Methods of altering plant stress response

Abstract

The invention relates to method of altering a plant stress response comprising contacting a plant or part thereof with a formulation. The invention also relates to genetically altered plants with an altered stress response and methods of producing said plants.

Description

[0001] Methods of altering plant stress response

[0002] Introduction

[0003] Biotic and abiotic stresses (such as salt, drought, heat, cold, heavy metals, ozone, UV radiation, and nutrient deficiencies) have detrimental effects on plant growth and yield. Plants respond in many ways to abiotic stresses, which allows them to tolerate and / or adapt to adverse conditions. Altering the response of plants to stress can increase plant yield, survival and productivity.

[0004] Minimising the impact of adverse environments on plant growth is of vital importance for agriculture and food security. Adverse environments were estimated to cause an overall yield loss of ~70% in key agricultural crops, i.e., the average yield was only ~30% of the genetic yield potential (Zhang et al (2020) Dev Cell 55(5), 529-443).

[0005] As an example, agricultural practices and poor irrigation management in warm and dry regions often result in saline and gypsiferous soils with a low productivity. Indeed, secondary salinization resulting from poor irrigation management affects approximately 20% of irrigated land worldwide. Thus, abiotic stressors such as salt stress represents a serious limitation to soil productivity. There is therefore a need to improve crop yields in soils subjected to salinity constraints and other abiotic stressors.

[0006] Dimethylsulfoniopropionate (DMSP) is one of Earth’s most abundant and ecologically important organosulfur molecules, with roles as an osmolyte1, cryoprotectant2, antioxidant3, baroprotectant4, grazing deterrent5, in chemotaxis6, global carbon and sulfur cycling, and, potentially, in climate regulation, since it is a major precursor for the climate-active gases dimethylsulfide (DMS) and methanethiol7. Marine algae and bacteria are major global DMSP producers8, but invasive perennial Spartina grasses9(e.g. S. anglica and S. alterniflora) also display high intracellular DMSP concentrations and are responsible for considerable DMSP production and cycling in saltmarshes. Per unit area, the DMSP concentrations produced by these plants are far higher than the oceans and are proposed to contribute ~10% of global atmospheric DMS emissions10. Only seagrasses such as Posidonia oceanica and Zostera marina^, Saccharum officinarum (sugarcane)12, and sea daisy Melanthera biflora (formerly Wollastonia biflora)Kproduce similarly high DMSP concentrations, though some other plants can produce DMSP at concentrations 1000 times lower14 15.

[0007] Analyses of DMSP have focused on marine phytoplankton and plants where it is found in higher concentrations, e.g. S. alterniflora. The role of DMSP in higher plants is unclear. It has been reported that in vitro application of DMSP to Paraquat-treated isolated leaf discs of Panicum commutatum in petri dishes has a protective effect (Husband et al., Environmental and Experimental Botany 79 (2012) 44-48).

[0008] The present invention is aimed at addressing the need for generating plants with improved stress responses. Summary of the Invention

[0009] The inventors have identified the first genes involved in DMSP synthesis in plants. They have shown that S. anglica DMSP synthesis enzymes are efficient and highly expressed compared to homologs from plants that accumulate low DMSP concentrations. They have demonstrated that DMSP has roles in oxidative and osmotic stress, that DMSP levels are manipulable in low-accumulation species and that doing so can increase tolerance to salinity and drought. This work provides the first molecular genetic understanding of DMSP production in plants, with the findings opening new research avenues to address the contribution of plants to global DMSP cycling and identifies a promising role for DMSP in plant biotechnology.

[0010] The inventors have shown that exogenous application of a formulation comprising DMSP to plants, e.g. application to the roots, leads to translocation and stable and long-term accumulation of DMSP in the leaves which is protective against stress.

[0011] The inventors have also shown that over-expression of S. anglica DMSP synthesis genes improves plant tolerance to stress such as to salinity and drought. The invention therefore provides plants with an altered stress response for use in plant industries, for example agriculture and / or horticulture, as well as methods of altering a plant stress response.

[0012] According to a first aspect of the invention, there is provided a method of altering a plant stress response comprising contacting a plant with a formulation comprising dimethylsulfoniopropionate (DMSP) wherein said plant is not Panicum commutatum.

[0013] In another aspect, the invention relates to a genetically altered plant wherein said plant expresses one or more nucleic acid from Spartina, a bacterium or / and algae involved in DMSP synthesis. Exemplary sequences are those listed in Tables 1 a, 1 b, 2 and the section entitled Listing of sequences or sequences with at least 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity thereto.

[0014] In another aspect, the invention relates to a genetically altered plant wherein said plant expresses a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

[0015] In another aspect, the invention relates to an isolated polypeptide comprising SEQ ID NO: 2 or a sequence having 90% sequence identity thereto.

[0016] In another aspect, the invention relates to an isolated polypeptide comprising SEQ ID NO: 4 or a sequence having 92% sequence identity thereto.

[0017] In another aspect, the invention relates to an isolated polypeptide comprising SEQ ID NO: 6 or a sequence having 90% sequence identity thereto.

[0018] In another aspect, the invention relates to an isolated nucleic acid sequence comprising SEQ ID NO: 1 or a sequence having 90% sequence identity thereto.

[0019] In another aspect, the invention relates to an isolated nucleic acid sequence comprising SEQ ID NO: 3 or a sequence having 92% sequence identity thereto.

[0020] In another aspect, the invention relates to an isolated nucleic acid sequence comprising SEQ ID NO: 5 or a sequence having 90% sequence identity thereto.

[0021] In another aspect, the invention relates to a vector comprising an isolated nucleic acid sequence as described herein.

[0022] In another aspect, the invention relates to a host cell comprising an isolated nucleic acid sequence or vector as described herein.

[0023] In another aspect, the invention relates to a use of a nucleic acid sequence or the vector as described herein for altering the stress response of a plant. In another aspect, the invention relates to method for altering the stress response of a plant, comprising introducing into said plant. a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto; and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

[0024] In another aspect, the invention relates to a method for producing a plant with an altered stress response, comprising introducing into said plant a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

[0025] In another aspect, the invention relates to a method of altering a plant stress response comprising contacting the roots of a plant with a formulation comprising DSMP. Contacting the roots leads to uptake of DSMP.

[0026] Figures

[0027] The invention is described in the following non-limiting figures.

[0028] Figure 1 : Spartina anglica produces DMSP via SDC and DOX a, Spartina anglica at Stiffkey saltmarsh, May 2021. b, DMSP synthesis pathway in S. anglica converts methionine to DMSP via four enzyme activities (green), c, HPLC trace of SDC assay for recombinant S. anglica SDC (SaSDC, S-methylmethionine decarboxylase) incubated with SMM, leading to production of DMSP-amine, alongside an SMM only control, d, GC result of DOX assay performed on enzyme extracts from S. anglica leaf tissue, recombinant S. anglica copper amine oxidase 1 (SaCAOl) and recombinant S. anglica DOX (SaDOX, DMSP-amine oxidase). Enzymes are incubated with DMSP-amine or DMSP-amine and competing substrate putrescine in excess, leading to production of DMSP-aldehyde that spontaneously degrades to produce DMS. Pig kidney diamine oxidase used as a positive control, e, RT-qPCR of S. anglica DMSP synthesis genes in denoted tissues, f, HPLC traces of SDC assays performed on enzyme extracts prepared from denoted S. anglica tissues. Retention times of SMM (6.5 min) and DMSP-amine (7.6 min) are indicated in blue and red, respectively, g, DOX assays performed on enzyme extracts prepared from denoted S. anglica tissues, normalised to 1 pg of total extracted protein and with activities shown relative to leaf, h, DMSP accumulation in denoted S. anglica tissues. Data represent mean ± one standard error from n>3 independent biological replicates. Letters indicate statistical significance after ANOVA followed by Tukey test; n.s. denotes no statistically significant difference.

[0029] Figure 2: SaMMTI is a functional methionine S-methyltransferase a, HPLC traces of recombinant SaMMTI , HvMMT, AtMMT incubated with methionine and SAM, demonstrating the production of SMM from methionine and SAM of all three enzymes. MBP control is purified maltose binding protein incubated with MMT assay reaction mix. b, Alignment of MMT from Zea mays against SaMMT sequences demonstrating insertion in SaMMT2 SAM binding domain. Red triangle indicates the beginning of the domain. ZmMMT refers to position 69 to 125 of SEQ ID NO: 8, SaMMTI refers to position 64 to 97 of SEQ ID NO: 6, SaMMT2 refers to position 66 to 101 of SEQ ID NO: 10.

[0030] Figure 3: SaODC has SDC activity, whereas SaDAPDC and SaADC do not a, HPLC traces of diaminopimelate retention time, and recombinant SaDAPDC incubated with diaminopimelate (DAP), demonstrating this enzyme is functional in vitro, b, HPLC traces shows SMM retention time, and incubation of SaDAPDC and SaADC with SMM, which show no activity with this substrate despite being in vitro functional enzymes, c, LC-MS total ion chromatogram of products of SaODC incubation with SMM, demonstrating presence of DMSP-amine (top) and expected mass of protonated form of DMSP-amine (bottom), d, Reaction rate of SaSDC with varying concentrations of SMM, used to calculate Kmand Vmax of the enzyme, e, NMR assignment of SMM (blue, lower) and DMSP-amine (red, upper). For quantification of the reaction rate in panel d, peak number three (corresponding to the CH2 adjacent to the sulphur) was used, f, NMR assignment of ornithine (red, middle) and putrescine (green, upper). No reduction is observed in peak 1 and 2 in the blue (lower) spectra, after 1 hour of incubation with 35 pg of SDC, suggesting limited activity on this substrate.

[0031] Figure 4: SaDOX and SaCAOl have major and minor DOX activities, respectively a, Fold enrichment of DOX activity in total protein extracts from S. anglica leaves following successive rounds of chromatography, b, SDS-PAGE gel of 3 pg S. anglica leaf protein fractions following each step of the activity tracking process, demonstrating removal of proteins with each round. Highlighted lanes are the samples which were sent for mass spectrometry analysis. AS: ammonium sulfate, PS: phenyl sepharose, Q: Q strong anion exchange, GF: gel filtration. Number following '#' refers to the fraction number after Akta separation. Figure 5: DMSP accumulation in Spartina anglica is associated with stress responses a, Satellite image of Spartina anglica sampling sites at Stiffkey saltmarsh, taken from Google Maps. b, Variability in DMSP accumulation in individual clumps of S. anglica harvested from the sites denoted in panel a. Samples identified as ‘highest accumulators’ (red) and ‘lowest accumulators’ (blue) were subjected to RNA-sequencing. c, Volcano plot demonstrating contrasting gene expression profiles between highest and lowest DMSP accumulating clumps of S. anglica. Each dot represents a gene, with red dots (upper right quadrant) indicating genes that are more highly expressed in highest accumulators, blue dots (upper left quadrant) indicating genes that are lower, and grey dots (upper middle quadrant, lower quadrants) denoting genes that are not significantly differentially expressed. Dotted lines indicate significance thresholds. Labelled dot refers to Ethylene Responsive Element Binding Protein 1 (EREBPT), a key stress responsive transcription factor in grasses, d, Gene ontology enrichment analysis of differentially expressed genes from panel c, demonstrating an enrichment of stress-responsive genes in highest DMSP accumulators, e, Heatmap of pairwise Pearson correlations between measured variables (abundance of indicated elements and amino acids in S. anglica leaves), demonstrating negative linear relationship between DMSP and levels of nitrogen and proline, and positive linear relationship between DMSP and molybdenum (Mo). Crosses indicate that a relationship is not significant. Data represent mean ± one standard error from n>3 independent biological replicates. Letters indicate statistical significance after ANOVA followed by Tukey test

[0032] Figure 6: DMSP accumulation in Spartina anglica a, Example clump of S. anglica growing at Siffkey, Norfolk, UK. b, PCA of RNA-seq libraries generated from highest and lowest DMSP accumulating S. anglica. Libraries cluster, but not based on DMSP accumulation, c, Heatmap of pairwise Pearson correlations between expression level of SaMMT, SaSDC and SaDOX and DMSP, methionine (Met) and SMM, showing that gene expression levels do not correlate with DMSP accumulation, but SaMMT expression does correlate with SMM accumulation, d, Heatmap of pairwise Pearson correlations as in Figure 2e but elements represent measurements from soil rather than leaf tissue. Amino acid measurements derive from leaf, as do DMSP measurements. This shows the relationship between nutrient and heavy metal levels in the soil and DMSP and amino acid accumulation in leaves. Crosses indicate that a relationship is not significant, e, Sulfate and O-acetylserine measurements in leaf tissues from highest and lowest DMSP-accumulating clumps of S. anglica. n.s. denotes no statistically significant difference in a two- tailed student’s t-test.

[0033] Figure 7: Spartina anglica produces high levels of DMSP due to its unique SDC activity a, DMSP measurements showing accumulation of the compound in a large diversity of different plant species, b, Phylogenetic tree of ODC / SDCs in plant species related to S. anglica. c, HPLC traces of SDC assays showing that the recombinant S. anglica SDC produces DMSP-amine from SMM, whilst enzyme homologs from other plant species do not have this activity, d, Phylogenetic tree of DOX homologs in plant species related to S. anglica e, DOX assays performed with selected enzyme homologs from S. anglica (Sa), Saccharum offinarum (So), Setaria viridis (Sv) and Solanum lycopersicum (SI), f, Number of reads mapping to homologs of denoted DMSP synthesis genes in each species, normalised to the number of reads mapping to ACT8 homologs (square rooted for clarity), showing levels of DOX are much higher in S. anglica, despite equivalent enzymology. EF1A is included as a reference gene control, to demonstrate expression differences of other genes are not attributable to read-depth artifacts. Data represent mean ± one standard error from n>3 independent biological replicates. Letters indicate statistical significance after ANOVA followed by Tukey test.

[0034] Figure 8: Other plant species have MMT enzymes, DOX activity and functional ODC enzymes a, Phylogenetic tree of MMT proteins from higher plants rooted against bacterial mmtN, showing that SaMMTI and SaMMT2 closely resemble other plant MMT enzymes, b, HPLC traces of recombinant SaSDC and homologous ODC enzymes incubated with ornithine, demonstrating all enzymes have in vitro functionality, and that SaSDC works less well with this substrate, c, DOX assays performed on protein extracts from the denoted species, demonstrating S. anglica has enhanced DOX activity per unit protein, and in all cases DOX enzymes are inhibited by addition of excess putrescine. Data represent mean ± one standard error from n>3 independent biological replicates.

[0035] Figure 9: Root uptake of DMSP or over-expression of S. anglica MMT, SDC and DOX have protective effects against salt stress and drought a, GC measurements of leaf tissue of tomato shown in Figure 10a, demonstrating that DMSP is taken up by roots and accumulates in leaf tissue, b, PCA plot of RNA-seq performed on tomato experiment presented in Figure 4, showing treatment groups cluster together, c, Volcano plot demonstrating broad transcriptomic changes in tomato following NaCI treatment. Each dot represents a gene, with blue dots (upper left quadrant) indicating genes that are downregulated in NaCI treatment, red dots (upper right quadrant) representing upregulation, and grey dots (upper middle quadrant, lower quadrants) denoting genes that are not significantly differentially expressed. Dotted lines indicate significance thresholds, d, Volcano plot as in panel c, but comparing DMSP treated vs untreated tomato plants, e, Image of N. benthamiana plants under drought conditions, in which every leaf has either been infiltrated with Agrobacterium tumefaciens containing no plasmid (control infiltrated) or containing a plasmid to over-express S. anglica MMT, SDC and DOX. f, Pie charts representing the frequency of wilted vs non wilted leaves from plants treated as described in panel e. Data represent mean ± one standard error from n>3 independent biological replicates.

[0036] Figure 10: DMSP protects plants from abiotic stress a, Image of representative tomato plants from each treatment group, demonstrating that biomass loss caused by NaCI is rescued by DMSP treatment, b, Fresh weight measurements of tomato plants from each treatment group, demonstrating that biomass loss caused by NaCI is rescued by DMSP treatment, c, Volcano plot demonstrating contrasting gene expression profiles of tomato plants treated with salt stress alone (NaCI) or salt stress and DMSP (NaCI + DMSP). Each dot represents a gene, with red dots (upper right quadrant) indicating genes that are more highly expressed in NaCI treatment, blue dots (upper left quadrant) representing downregulation, and grey dots (upper middle quadrant, lower quadrants) denoting genes that are not significantly differentially expressed. Dotted lines indicate significance thresholds, d, Clustered heatmap of average normalised count numbers of differentially expressed genes of interest from panel c, showing that samples from Control and NaCI + DMSP-treated plants cluster together away from samples from NaCI-treated plants, e, DMSP measurements of Nicotiana benthamiana leaves transformed with different S. anglica DMSP synthesis genes, showing that all three genes (MMT, SDC and DOX) are needed for high DMSP accumulation, f, DMSP measurements of Arabidopsis thaliana over-expressing S. anglica MMT, SDC and DOXshowing highly elevated accumulation in four independent transgenic lines, g, Images of wildtype (WT, Col-0) and T2 transgenic A. thaliana seedlings (Line 2) growing in the presence and absence of NaCI, demonstrating that over-expression of S. anglica DMSP synthesis genes (MMT, SDC and DOX) conveys tolerance to salt stress. Scale bar = 1 cm. h, Fresh weight measurements of wildtype and T3 transgenic plants, demonstrating that biomass rescue is statistically significant (n=36-54 plants per condition, with at least 4 technical replicates per condition). All other data represent mean ± one standard error from n>3 independent biological replicates. Letters indicate statistical significance after ANOVA followed by Tukey test; n.s. denotes no statistically significant difference.

[0037] Figure 11 : DMSP protects plants from salt stress and osmotic stress

[0038] Fresh weight measurements of tomato plants from each treatment group grown in agar, demonstrating that biomass loss caused by salt stress (100 mM NaCI) or osmotic stress (200 mM sorbitol) is increased by DMSP treatment (5 pM DMSP). The percentage increase in total biomass (root and shoot) of plants treated with stress and DMSP is indicated relative to plants treated with the respective stress alone. Data represent mean ± one standard error from n>4 biological replicates. Letters indicate statistical significance in two-tailed t-test (p<0.05).

[0039] Figure 12: Watering of plants and uptake of DMSP through the roots leads to long lasting systemic effects

[0040] Watering tomato plants in soil with a single dose of a solution containing the indicated concentrations of DMSP (50 or 500 pM) leads to accumulation of DMSP in the soil for up to c. 2 weeks after watering (a), but long lasting accumulation of DMSP in the leaves (b). No visual negative impacts on plant growth were observed in response to watering with DMSP (c). Data represent mean ± one standard error from n>3 biological replicates.

[0041] Figure 13: Tomato plants watered with DMSP show improved tolerance to salt stress a, Image of representative tomato plants from each treatment group, demonstrating that plants growing in soil and watered with a single dose of a solution containing 50 pM DMSP and then supplemented with 100 mM NaCI are larger than plants supplemented with 100 mM NaCI alone (i.e. with no DMSP pre-treatment). Scale bar = 5 cm. b, Image of representative leaves (leaf 3) from tomato plants from each treatment group in (a). Scale bar = 2 cm. c, Total chlorophyll content measured in leaves from each treatment group in (b). Data represent mean ± one standard error from n>3 biological replicates. Letters indicate statistical significance in two-tailed t-test (p<0.05).

[0042] Figure 14: Spray application of DMSP improves plant tolerance to oxidative stress

[0043] Application of 500 pM DMSP to tomato plants (applied as a single spray dose to leaves, three days before stress treatment) improves tolerance to oxidative stress (induced by 1 mM Paraquat treatment to leaves for 3 days) as measured by total chlorophyll content. Stars indicate statistical significance in two-tailed t-test (p<0.05).

[0044] Figure 15: Transgenic barley plants produce high concentrations of DMSP a, Transgenic barley plants overexpressing MMT, SDC and DOX produce high concentrations of DMSP (green bars), with some lines (red bars) producing DMSP at concentrations 10-100 fold greater than Spartina anglica (grey bar), b, Image of representative transgenic barley plants overproducing DMSP reveal no visual negative impacts on plant growth are observed.

[0045] Figure 16: Sulfate transporters are expressed in Spartina anglica a, Phylogenetic tree of SULTRs from Arabidopsis thaliana (black) and S. anglica (orange), b, RT- qPCR of selected S. anglica SULTR genes in denoted tissues, with values for leaf tissue normalised to 1 for each gene. SULTR3;7 expression was not detected (n.d.) in rhizome or root tissue. Data represent mean ± one standard error from n=4 independent biological replicates, c, Heatmap of read counts for SULTR genes in leaf tissues from highest and lowest DMSP-accumulating clumps of S. anglica, with values representing Log2(read count +1) where 1 is a psuedocount. d, DMSP accumulation in glasshouse-grown S. anglica plants watered with deionised water (control), 500 mM NaCI, or 35 PSU seas salts. Data represent mean from six independent biological replicates, with three technical replicate measurements made per sample for each time-point (n=18). Letters indicate statistical significance after ANOVA followed by Tukey test.

[0046] Figure 17: Abundance of dddand dmdA genes in the sediments of Spartina alterniflora (high DMSP producer) and S. patens (low DMSP producer). Relative abundance of the nine prokaryotic DMSP lyase ddd genes and the DMSP demethylation dmdA gene. Data represent mean ± one standard error from three independent biological replicates (n=3). n.s. denotes no statistically significant difference after ANOVA followed by Tukey test.

[0047] Figure 18 DMSP biosynthesis genes, enzymes and pathways, a, The methylation’ pathway in some higher plants with the methionine (Met) S-methyltransferase (MMT) and bacteria containing MmtN or another methyltransferase (BurB) (left); the ‘transamination’ pathway in algae, bacteria and corals with DSYB / DsyB, DsyGD / DsyG, DSYE and / or TpMMT (middle); and the ‘decarboxylation’ pathway in Crypthecodinium cohnii (right). The pathways are named after their first reaction step (in larger font). AdoMet, S-adenosylmethionine; AdoHcy, S-adenosylhomocysteine; NADP, nicotine adenine dinucleotide phosphate; MAT, methionine aminotransferase; MR, MTOB reductase; MSM, MTHB S-methyltransferase; DDC, DMSHB decarboxylase; SMM, S-methylmethionine; MTOB, 4- methylthio-2-oxybutyrate; MTHB, 4-methylthio2-hydroxybutyrate; DMSHB, 4-dimethylsulfonio-2- hydroxy butyrate; MTPA, 3-methylthiopropylamine; MMPA, 3-methylmercaptopropionate. Plant genes from Spartina anglica are MMT, SDC, DOX and ALDH.

[0048] Figure 19: DMSP supplementation in soil leads to long-lasting increases in DMSP concentrations in the leaf. Single root drench applications of the indicated concentrations of DMSP were made at day 0 in Solanum lycopersicum (tomato; panel a) and Hordeum vulgare cv. Golden Promise (barley; panel b). Plants were grown in controlled environment rooms.

[0049] Figure 20: Spray application of DMSP to barley leads to long-lasting increases in DMSP concentrations in the leaf. A single spray application of DMSP solution at the indicated concentrations was made to Hordeum vulgare cv. Golden Promise (barley) plants at week 0 and grown in controlled environment rooms. DMSP concentrations was monitored in non-sprayed leaf tissue at the indicated timepoints after the initial spray application. Independent replicates are shown in panels a and b.

[0050] Figure 21 : Spray application of DMSP to barley allows plants to tolerate salt stress. A single spray application of DMSP solution at the indicated concentrations was made to Hordeum vulgare cv. Golden Promise (barley) plants grown in controlled environment rooms. Plant biomass (dry weight, DW) was measured after 15 weeks of growth under normal conditions (watering with fresh water; panel a) or salt stress (watering with 100 mM NaCI; panel b). Stars indicate significant difference (p<0.05) relative to control samples.

[0051] Figure 22: Spray application of DMSP to pea allows plants to tolerate drought stress. One spray application of a 50 mM DMSP solution was made to Pisum sativum cv. Kelvedon Wonder (pea) plants grown under controlled environment conditions. Plants were subjected to drought stress by decreased watering regime (50% watering relative to normal conditions). Plant biomass (fresh weight, FW; panel a), chlorophyll content (panel b) and DMSP concentration in non-sprayed leaves (panel c) was measured after 8 weeks of growth. Plant phenotypes of 8-week-old plants are shown (panel d). Stars indicate significant difference (p<0.05) relative to control samples, n.s. Not significant.

[0052] Figure 23: Root drench and spray applications of DMSP to wheat both lead to long-lasting increases in DMSP concentrations in the leaf in the glasshouse, a, Three spray applications or three root drench applications of DMSP solutions (given three weeks apart) at the indicated concentrations were made to Triticum aestivum cv. Bamford (wheat) plants grown under glasshouse conditions. DMSP concentrations was monitored in non-sprayed leaf tissue at the indicated timepoints after the second application (week 0). b, Chlorophyll content was measured in plants 7 weeks after the second DMSP application. Stars indicate significant difference (p<0.05) relative to control (untreated) samples.

[0053] Figure 24: Spray application of DMSP to wheat allows plants to tolerate drought stress. Three spray applications of DMSP solutions (given three weeks apart) at the indicated concentrations were made to Triticum aestivum cv. Bamford (wheat) plants grown under glasshouse conditions. Plants were subjected to drought stress by decreased watering regime (50% watering relative to normal conditions). Plant biomass (fresh weight, FW; panel a) and chlorophyll content (panel b) was measured after ~29 weeks of growth. Stars indicate significant difference (p<0.05) relative to control samples. Bars represent mean ± SE from three independent replicates (n=61-74).

[0054] Figure 25: Spray application of DMSP to wheat leads to long-lasting increases in DMSP concentrations in the leaf in the field. Two or three spray applications of DMSP solutions (given approximately three weeks apart) at the indicated concentrations were made to Triticum aestivum cv. Bamford (wheat) plants grown in the field (Yorkshire, UK). DMSP concentrations was monitored in non-sprayed leaf tissue 9 weeks after the final application.

[0055] Figure 26: Transgenic barley plants are more tolerant to drought stress. Three independent transgenic Hordeum vulgare cv. Golden Promise (barley) lines demonstrate decreased wilting relative to wildtype (non-transgenic) plants. Plants were subjected to drought stress by decreased watering regime (50% watering relative to normal conditions) and were grown in controlled environment rooms.

[0056] Detailed Description

[0057] The embodiments of the invention will now be further described. In the following passages, different embodiments are described. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous. The practice of the present invention will employ, unless otherwise indicated, conventional techniques of botany, microbiology, tissue culture, molecular biology, chemistry, biochemistry and recombinant DNA technology, bioinformatics which are within the skill of the art. Such techniques are explained fully in the literature.

[0058] Generally, nomenclatures used in connection with, and techniques of, cell and tissue culture, plant biology, molecular biology, biochemistry, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art. The methods and techniques of the present disclosure are generally performed according to conventional methods well-known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. See, e.g., Green and Sambrook et al., Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2012).

[0059] Method of altering a plant stress response The inventors have shown that exogenous application of DMSP, e.g. to roots or leaves, improves the tolerance of plants to stress.

[0060] Therefore, according to a first aspect of the invention, there is provided a method of altering a plant stress response comprising contacting a plant with a formulation comprising dimethylsulfoniopropionate (DMSP) wherein said plant is not Panicum commutatum. In one aspect, the plant is not a Panicum species.

[0061] In one embodiment, the term “altering” means “increasing”, “improving” or “enhancing.” Altering the stress response can be an improvement of plant stress tolerance of at least 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% or more in comparison to a control plant, or 2-fold, 3-fold, 4-fold, 5-fold, 6- fold, 7-fold, 8-fold, 9-fold or 10-fold or more in comparison to a control plant. Thus, the methods described herein improve a plants’ tolerance to stress. Improving the stress response leads to an increase in yield. An improvement in the stress response can be measured by measuring yield.

[0062] Various methods are available in the art to measure the tolerance of plants, some of which are described in the examples here below. Increased stress tolerance will usually be apparent from the general appearance of the plants and may be measured e.g., by increased biomass production, continued vegetative growth under adverse conditions or higher seed yield. Stress tolerant plants have a broader growth spectrum, i.e. they are able to withstand a broader range of climatological and other abiotic changes, without yield penalty, as compared to control plants. Biochemically, stress tolerance may be apparent as the higher NAD+-NADH / ATP content and lower production of reactive oxygen species of stress tolerant plants compared to control plants under stress condition. Stress tolerance may also be apparent as the higher chlorophyll content, higher germination rates, higher photosynthesis and lower chlorophyll fluorescence under stress conditions in stress tolerant plants compared to control plants under the same conditions.

[0063] It will be clear that it is also not required that the plant be grown continuously under the adverse conditions for the stress tolerance to become apparent. Usually, the difference in stress tolerance between a plant or plant cell produced according to the invention and a control plant or plant cell will become apparent even when only a relatively short period of adverse conditions is encountered during growth.

[0064] The term "yield" in general means a measurable produce of economic value, typically related to a specified crop, to an area, and to a period of time. Individual plant parts directly contribute to yield based on their number, size and / or weight, or the actual yield is the yield per square meter for a crop and year, which is determined by dividing total production (includes both harvested and appraised production) by planted square meters. The term "yield" of a plant may relate to vegetative biomass (root and / or shoot biomass), to reproductive organs, and / or to propagules (such as seeds) of that plant. Thus, according to the invention, yield comprises one or more of and can be measured by assessing one or more of: increased seed yield per plant, increased seed filling rate, increased number of filled seeds, increased harvest index, increased number of seed capsules and / or pods, increased seed size, increased growth or increased branching, for example inflorescences with more branches or increased biomass. Yield is increased relative to control plants, for example by at least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more, or 1.5-fold, 1.75-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold or 10-fold or more.

[0065] According to this aspect, a control plant is defined as a plant that has not been exposed to DMSP. Preferably, the control plant is of the same species.

[0066] In one embodiment of the method, the plant is not of the genus Panicum.

[0067] The term plant as used in accordance with this aspect does not include isolated plants parts, such as cuttings. The term isolated refers to parts of the plant that have been removed from the plant, e.g. isolated leaves or parts thereof.

[0068] The term plant part in connection with this aspect of the invention includes leaves, shoots, stems, flowers, fruits and roots.

[0069] Preferably, the term plant part as used in accordance with this aspect is not an isolated plant part and the method is not carried out in vitro, e.g. by applying the formulation to the isolated plant part in vitro.

[0070] For example, the term plant part does not refer to isolated leaves or parts thereof, e.g. isolated leaf discs.

[0071] In one embodiment, the plant produces no endogenous DMSP or low levels of endogenous DMSP. In one embodiment the plant produces 20 nmol g-1FW (fresh weight) of endogenous DMSP. In one embodiment the plant produces up to 20 nmol g-1FW of endogenous DMSP. In one embodiment the plant produces up to 1 nmol g-1FW of endogenous DMSP, up to 2 nmol g-1FW of endogenous DMSP, up to 3 nmol g-1FW of endogenous DMSP, up to 4 nmol g-1FW of endogenous DMSP, up to 5 nmol g-1FW of endogenous DMSP, up to 6 nmol g-1FW of endogenous DMSP, up to 7 nmol g-1FW of endogenous DMSP, up to 8 nmol g-1FW of endogenous DMSP, up to 9 nmol g-1FW of endogenous DMSP, up to 10 nmol g-1FW of endogenous DMSP, up to 11 nmol g-1FW of endogenous DMSP, up to 12 nmol g-1FW of endogenous DMSP, up to 13 nmol g-1FW of endogenous DMSP, up to 14 nmol g-1FW of endogenous DMSP, up to 15 nmol g-1FW of endogenous DMSP, up to 16 nmol g-1FW of endogenous DMSP, up to 17 nmol g-1FW of endogenous DMSP, up to 18 nmol g-1FW of endogenous DMSP, or up to 19 nmol g-1FW of endogenous DMSP. In one embodiment, the plant produces 5 nmol g-1FW of endogenous DMSP. In one embodiment, the plant produces up to 1 nmol g-1FW of endogenous DMSP, up to 2 nmol g-1FW of endogenous DMSP, up to 3 nmol g-1FW of endogenous DMSP, up to 4 nmol g-1FW of endogenous DMSP, up to 5 nmol g-1FW of endogenous DMSP, or up to 6 nmol g-1FW of endogenous DMSP.

[0072] The inventors have surprisingly shown that when DMSP is applied to a plant, it is taken up by the plant and, from just one application, DMSP concentrations in the plant remain elevated for a prolonged time, e.g. several months. When the plant is contacted with a formulation comprising DMSP by soil supplementation, the DMSP is translocated to and accumulates in the leaves after being taken up by the roots. Thus, following uptake of DMSP through the roots, increased DMSP concentration was observed in the leaves.

[0073] This demonstrates that when DMSP is applied to a plant, e.g. by watering the plant and uptake of DMSP through the roots, this leads to long lasting systemic effects.

[0074] This is advantageous so as prophylactically protect against stresses e.g. drought protection which can occur without much warning. Protection against future stresses can therefore be applied in advance before the emergence of any stress to protect plants and yield.

[0075] In one embodiment, the formulation is a liquid composition. In one embodiment, the liquid composition is an aqueous solution. In one embodiment, the formulation may include algae material. Optionally, the algae material may be fractionated using routine methods known in the art so as to as to increase the purity of DMSP containing fractions. Therefore, in one embodiment, the formulation may comprise fractionated algae, with DMSP as an active ingredient. In one embodiment, the formulation does not comprise fractionated algae.

[0076] In one embodiment, contacting the plant is by application to the roots.

[0077] In one embodiment, contacting the plant, e.g. application to the roots, may comprise watering. Thus, one embodiment, the method of the invention may comprise topically applying the formulation, and / or watering the plant with the formulation.

[0078] In one embodiment, watering the plant may comprise fertigation, hand watering, soil drenching soil supplementation or irrigation. Different types of irrigation systems can be used, including surface irrigation, localized irrigation, drip irrigation, sprinkler irrigation, center pivot irrigation, lateral move irrigation, sub-irrigation or manual irrigation.

[0079] The method for the invention also extends to foliar application. Foliar application, also known as “foliar nutrition” or “foliar fertilization” or “foliar feeding” is a process of applying nutrients or formulations to above-ground plant parts in order to supply the plant with nutrients or formulations which are traditionally applied to the soil and taken up by the roots. The formulation enters the plant via the cuticle or stomata before entering the plant cell.

[0080] Thus, one embodiment, topically applying may comprise foliar feeding. Thus, in one embodiment foliar feeding may comprise spraying. The topical application is onto a plant body for example spraying on leaves, flowers, or fruits.

[0081] In one embodiment, the plant or part thereof is contacted once with an application of DMSP formulation before harvest. For example, in one embodiment, the plant or part thereof is sprayed once with an application of DMSP formulation before harvest. In one embodiment, the plant or part thereof is contacted more than once with an application of DMSP formulation before harvest. For example, in one embodiment, the plant or part thereof is sprayed more than once with an application of DMSP formulation before harvest. In one embodiment, the plant or part thereof is contacted 3 times with an application of DMSP formulation before harvest. In one embodiment, the plant or part thereof is contacted once, twice, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times or more with an application of DMSP formulation before harvest. For example, in one embodiment, the plant or part thereof is sprayed 3 times with an application of DMSP formulation before harvest. In one embodiment, the plant or part thereof is sprayed once, twice, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times or more with an application of DMSP formulation before harvest. In one embodiment, the plant or part thereof is contacted every about 3 weeks with an application of DMSP formulation before harvest. In one embodiment, the plant or part thereof has about 3 weeks gap between contact or spray with DMSP. In one embodiment, the plant or part thereof has about 1 weeks gap, about 2 weeks gap, about 3 weeks gap, about 4 weeks gap, about 5 weeks gap, about 6 weeks gap, about 7 weeks gap, about 8 weeks gap or more between contact or spray with DMSP. Thus, in one embodiment, the plant or part thereof is sprayed 3 times with an application of DMSP formulation before harvest, and has about 3 weeks gap between spray with DMSP. Thus, in one embodiment, the plant or part thereof is contacted 3 times with an application of DMSP formulation before harvest, and has about 3 weeks gap between contact with DMSP.

[0082] In one embodiment, the plant may be a dicot or monocot plant. In one embodiment the dicot or monocot plant may be selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco, grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut barley or oats. In one embodiment the plant may be selected from Solanum lycopersicum, Arabidopsis thaliana, Nicotiana benthamiana, or Hordeum vulgare.

[0083] In one embodiment, the plant is a plant used in agriculture. In one embodiment, the plant is a crop plant. By crop plant is meant any plant which is grown on a commercial scale for human or animal consumption or use. In one embodiment, the plant is a cereal. In another embodiment, the plant is selected from rice (Oryza sativa), maize (Zea mays), wheat (Triticum aestivum), sorghum (Sorghum bicolor, Sorghum vulgare), brassica, soybean and millet. In one embodiment, the plant is selected from rice, such as the japonica or indica varieties. Other exemplary genetically altered plants of the invention include, but are not limited to, canola (Brassica napus, Brassica rapa ssp., Brassica Oleracea), alfalfa (Medicago sativa), rape (Brassica napus), rye (Secale cereale), sunflower (Helianthus annuus), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium hirsutum), sweet potato (Ipomoea batatas), cassava (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp), avocado (Persea americana), fig (Ficus carica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), apple (Malus domestica), blackberry (Rubus), strawberry (Fragaria), walnut (Juglans regia), grape (Vitis vinifera), apricot (Prunus armeniaca), cherry (Prunus), peach (Prunus persica), plum (Prunus domestica), pear (Pyrus communis), watermelon (Citrullus vulgaris), duckweed (Lemna), oats, barley, vegetables, ornamentals, conifers, and turfgrasses (e.g., for ornamental, recreational or forage purposes), Cannabis sativa, Cannabis indica, Pennycress (Thlaspi spp.) and biomass grasses (e.g., switchgrass and miscanthus). In one embodiment, the plant is a plant used in horticulture. By horticulture, it is meant smaller scale plant management including the management of garden crops, fruits, vegetables, and ornamental plants.

[0084] The term “plant stress” means a disadvantageous impact on the physiology of a plant, which may be induced upon a departure from optimal environmental conditions. Plants are considered to be under stress when environmental conditions are not ideal for growth.

[0085] In one embodiment, the stress is abiotic or biotic stress.

[0086] Biotic stress includes pathogen or pest infection including infection by fungi, viruses, bacteria, insects, nematodes, mycoplasms and mycoplasma like organisms, etc. Abiotic stress is caused by deficiencies or excesses in environmental factors including water, salt, light, temperature, and nutrients can substantially reduce plant growth and productivity and even survival. Abiotic stress reduces plant growth because when plants are under stress, the environmental conditions are not optimal for growth processes such as photosynthesis and protein synthesis, resulting in decreased plant growth and biomass production. Abiotic stressors may promote the generation of reactive oxygen species in photosynthetic cells, and cell death from abiotic stress may therefore be in part a result of oxidative damage.

[0087] Drought stress, for example, inhibits plant growth because water is needed for cell turgor that drives cell expansion. Slower plant growth under stress, however, is not only a passive consequence of the adverse environment. Plants under stress also actively slow their growth in order to adapt to the stress conditions. This “active” growth inhibition is achieved through stress-triggered cell signalling. The effect of stress on plant growth can be measured for example as a decrease in plant growth rate or as a decrease in biomass accumulation. An improved stress response therefore leads to increased yield, growth rate / and or biomass.

[0088] In one embodiment, the stress is abiotic stress. For example, the stress is osmotic stress, oxidative stress, heavy metal stress, drought, submergence, cold, UV radiation, heat, ozone, nutrient deficiency and / or salt stress. In one embodiment, the stress may be osmotic stress, herbicides, drought, and / or extremes of temperature. In one embodiment, more than one stress response may be altered.

[0089] In one embodiment, the liquid composition is an aqueous solution. In one embodiment, the concentration of DMSP in the formulation of the invention may be about 500pM. In one embodiment, the concentration of DMSP in the formulation may be about 10 pM, about 20 pM, about 30pM, about 40pM, about 50pM, about 100pM, about 150pM, about 200pM, about 250pM, about 300pM, about 350pM, about 400pM, about 450pM, about 500pM, about 550pM, about 600pM, about 650pM, about 700pM, about 750pM, about 800pM, about 850pM, about 900pM, about 950pM, about 1 M. In one embodiment, the formulation of the invention may be applied about 260pM, about 270pM, about 280pM, about 290pM, about 300pM, about 31 OpM, about 320pM, about 330pM, about 340pM, about 350pM, about 360pM, about 370pM, about 380pM, about 390pM, about 400pM, about 41 OpM, about 420pM, about 430pM, about 440pM, about 450pM, about 460pM, about 470pM, about 480pM, about 490pM, about 500pM, about 51 OpM, about 520pM, about 530pM, about 540pM, about 550pM, about 560pM, about 570pM, about 580pM, about 590pM, about 600pM, about 61 OpM, about 620pM, about 630pM, about 640pM, about 650pM, about 660pM, about 600pM, about 61 OpM, about 620pM, about 630pM, about 640pM, about 650pM, about 660pM, about 670pM, about 680pM, about 690pM, about 700pM, about 710pM, about 720pM, about 730pM, about 740pM or about 750pM. In one embodiment, the formulation of the invention may be applied about 1 mM, 1 .1 mM, about 1 ,2mM, about 1 ,3mM, about 1 ,4mM, about 1 ,5mM, about 1 ,6mM, about 1 ,7mM, about 1 ,8mM, about 1 ,9mM, about 2mM, about 2.1 mM, about 2.2mM, about 2.3mM, about 2.4mM, about 2.5mM, about 2.6mM, about 2.7mM, about 2.8mM, about 2.9mM, about 3mM, 3.1 mM, about 3.2mM, about 3.3mM, about 3.4mM, about 3.5mM, about 3.6mM, about 3.7mM, about 3.8mM, about 3.9mM, about 4mM, about 4.1 mM, about 4.2mM, about 4.3mM, about 4.4mM, about 4.5mM, about 4.6mM, about 4.7mM, about 4.8mM, about 4.9mM, about 5mM, about 6mM, about 7mM, about 8mM, about 9mM, about 10mM, about 20mM, about 30mM, about 40mM, about 50mM, about 60mM, about 70mM, about 80mM, about 90mM, about 100mM, about 200mM, about 300mM, about 400mM, about 500mM, about 600mM, about 700mM, about 800mM, about 900mM, about 1 M. In one embodiment, the formulation of the invention may be applied about 1.1 M, about 1 ,2M, about 1 ,3M, about 1 ,4M, about 1 ,5M, about 1 ,6M, about 1 ,7M, about 1 ,8M, about 1 ,9M, about 2M, about 2.1 M, about 2.2M, about 2.3M, about 2.4M, about 2.5M, about 2.6M, about 2.7M, about 2.8M, about 2.9M, about 3M, 3.1 M, about 3.2M, about 3.3M, about 3.4M, about 3.5M, about 3.6M, about 3.7M, about 3.8M, about 3.9M, about 4M, about 4.1 M, about 4.2M, about 4.3M, about 4.4M, about 4.5M, about 4.6M, about 4.7M, about 4.8M, about 4.9M, about 5M.

[0090] In one embodiment, the formulation comprises further excipients, for example other agricultural products, such as fertilisers or pesticides. The term pesticide as used herein is to be understood broadly and encompasses any herbicide, fungicide, insecticide or mixtures thereof, i.e. the present disclosure is not limited to a specific kind of pesticide product. In one embodiment the excipients may be a liquid fertiliser such as nitrogen fertilisers, phosphate fertilisers, potassium fertilisers, calcium fertilisers, sulphur fertilisers and amendments, micronutrients, boric acid and leonardite, and combinations thereof, and / or in combination with one or more biostimulants selected from the group consisting of amino acid hydrolysates, humic extracts, algae extracts, live micro-organisms or extracts of micro-organisms, e.g. Pichia guilliermondii and Azotobacter chroococcum, and combinations thereof.

[0091] In one embodiment, the formulation does not comprise further excipients, for example other agricultural products, such as fertilisers or pesticides.

[0092] In the context of the invention, the term "fertiliser" refers to any organic or inorganic composition, natural or synthetic, which provides plants with one or more of the nutritional elements essential for the normal plant development thereof, usually primary macroelements (N, P, K), secondary macroelements (Ca, Mg, S) and microelements (B, Cl, Co, CU, Fe, Mn, Mo and Zn). Any suitable fertiliser may be used. The fertiliser is not considered to be a limitation to the invention.

[0093] The term herbicide as used herein is to be understood broadly in the present case and presents any herbicide material to be applied onto an agricultural field. Herbicides can specifically be referred to as selective or non-selective herbicides. A selective herbicide controls specific weed species, while leaving the desired crop relatively unharmed. In contrast, non-selective herbicides, e.g. called total weed killers, kill all plant material with which they come into contact. A herbicide may be at least one of the following, but is not limited thereto: acetamides, amides, aryloxyphenoxypropionates, benzamides, benzofuran, benzoic acids, benzothiadiazinones, bipyridylium, carbamates, chloroacetamides, chlorocarboxylic acids, cyclohexanediones, dinitroanilines, dinitrophenol, diphenyl ether, glycines, imidazolinones, isoxazoles, isoxazolidinones, nitriles, N- phenylphthalimides, oxadiazoles, oxazolidinediones, oxyacetamides, phenoxycarboxylic acids, phenylcarbamates, phenylpyrazoles, phenylpyrazolines, phenylpyridazines, phosphinic acids, phosphoroamidates, phosphorodithioates, phthalamates, pyrazoles, pyridazinones, pyridines, pyridinecarboxylic acids, pyridinecarboxamides, pyrimidinediones, pyrimidinyl(thio)benzoates, quinolinecarboxylic acids, semicarbazones, sulfonylaminocarbonyltriazolinones, sulfonylureas, tetrazol inones, thiadiazoles, thiocarbamates, triazines, triazinones, triazoles, triazolinones, triazolocarboxamides, triazolopyrimidines, triketones, uracils, ureas. Further, a herbicide may be, but are not limited thereto, lipid biosynthesis inhibitors, acetolactate synthase inhibitors (ALS inhibitors), photosynthesis inhibitors, protoporphyrinogen-IX oxidase inhibitors, bleacher herbicides, enolpyruvyl shikimate 3-phosphate synthase inhibitors (EPSP inhibitors), glutamine synthetase inhibitors, 7,8- dihydropteroate synthase inhibitors (DHP inhibitors), mitosis inhibitors, inhibitors of the synthesis of very long chain fatty acids (VLCFA inhibitors), cellulose biosynthesis inhibitors, decoupler herbicides, auxinic herbicides, auxin transport inhibitors, and / or other herbicides selected from the group consisting of bromobutide, chlorflurenol, chlorflurenol-methyl, cinmethylin, cumyluron, dalapon, dazomet, difenzoquat, difenzoquat- metilsulfate, dimethipin, DSMA, dymron, endothal and its salts, etobenzanid, flamprop, flamprop-isopropyl, flamprop-methyl, flamprop-M-isopropyl, flamprop-M- methyl, flurenol, flurenol-butyl, flurprimidol, fosamine, fosamine-ammonium, indanofan, indaziflam, maleic hydrazide, mefluidide, metam, methiozolin, methyl azide, methyl bromide, methyl-dymron, methyl iodide, MSMA, oleic acid, oxaziclomefone, pelargonic acid, pyributicarb, quinoclamine, tetflupyrolimet, triaziflam, tridiphane, and their agriculturally acceptable salts, amides, Isoxaflutole, Flufenacet, S-Metolachlor, Pendimethalin, Acetochlor, Pyroxasulfone, Cloransulam-methyl, Imazamethayr, Dimethenamid-P, Metamitrion, Ethofumesate, Quimerac, Prosulfocarb, Chlortoluron, Cinmethylin, Pendimethalin, esters or thioesters. Any suitable herbicide may be used. The herbicide is not considered to be a limitation to the invention.

[0094] As used herein, the term "fungicide" refers to the ability of a substance to decrease or inhibit growth of fungi or oomycetes. Any suitable fungicide may be used. The fungicide is not considered to be a limitation to the invention. Insecticides are pesticides used to kill insects. They include ovicides and larvicides used against insect eggs and larvae, respectively. Any suitable insecticide may be used. The insecticide is not considered to be a limitation to the invention.

[0095] The formulation may further comprise a plant growth regulator and / or plant health promoter biostimulant.

[0096] The formulation may optionally include one or more additional compounds providing an additional beneficial or otherwise useful effect. Such compounds include, without limitation, an adhesive, a surfactant, a solvent, a bio-agent, an emulsifying agent, a carrier, an adjuvant, a diluent, a dispersing agent, an insecticide, a pesticide, a fungicide, a fertilizer of a micronutrient or macronutrient nature, a herbicide, a feeding inhibitor, an insect moulting inhibitor, an insect mating inhibitor, an insect maturation inhibitor, a nematicide, a nutritional or horticultural supplement, or any combination thereof. In an aspect, a formulation described herein is odor free.

[0097] The formulation may optionally contain an agriculturally acceptable carrier. The agriculturally acceptable carrier is not particularly limited, and may be any of a solid carrier, a liquid carrier, and a gaseous carrier.

[0098] Examples of the solid carrier include clay, silicate, silica, resin, wax, fertilizer and the like. Examples of the liquid carrier include water, alcohol, ketone, petroleum solvent, saturated or unsaturated hydrocarbon, chlorinated hydrocarbon, liquefied petroleum gas and the like. Examples of the gaseous carrier include LPG, air, nitrogen, carbon dioxide, dimethyl ether and the like.

[0099] In combination with the active ingredients, the formulation may also comprise other compatible components which are, for example, a solid or liquid filler or diluent, an adjuvants, a preservative, a surfactant, a dispersant, a fixing agent, a defoaming agent, an anti-freezing agent, a dye, a thickening agent, an adhesive, a protective colloid, a penetrating agent, a stabilizer, a metal ion blocking agent, an aggregation preventing agent, an anti-rust agent, a pigment or other auxiliary agent, suitable for the desired use and which are acceptable for uses in agriculture. The formulations can be of any type known in the sector which are suitable for application onto all types of crops.

[0100] In one embodiment, the method of the invention generates a plant with elevated DMSP levels in the leaves or in other plant tissues such as roots, flowers, or fruits as compared to a plant that has not been contacted with DMSP. Elevated DMSP levels are elevated by at least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60, 70%, 80%, 90%, 100% or at least 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7- fold, 8-fold, 9-fold or 10-fold or more compared to a non-DMSP treated control plant. In one embodiment, the elevated DMSP levels last for at least 30 days. In one embodiment, the elevated DMSP levels last for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days, at least 25 days, at least 26 days, at least 27 days, at least 28 days, at least 29 days, at least 30 days, at least 31 days, at least 32 days, at least 33 days, at least 34 days, at least 35 days, at least 36 days, at least 37 days, at least 38 days, at least 39 days, at least 40 days, at least 41 days, at least 42 days, at least 43 days, at least 44 days, at least 45 days, at least 46 days, at least 47 days, at least 48 days, at least 49 days, at least 50 days. In one embodiment, the elevated DMSP levels last for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks. In one embodiment, the elevated DMSP levels last for at least 1 month, at least 1 .5 months, at least 2 months, at least 2.5 months, at least 3 months, at least 3.5 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months or at least 12 months.

[0101] The elevated DMSP levels may be measured by any one of suitable techniques in the art.

[0102] In another aspect, the invention also relates to a method of altering a plant stress response comprising contacting the roots of a plant with a formulation comprising DMSP. Contacting the roots leads to uptake of DMSP. Contacting the roots can be by means as described above, including irrigation.

[0103] Genetically altered plant

[0104] In another aspect, the invention relates to a genetically altered plant wherein said plant expresses one or more nucleic acid from Spartina, a bacterium or / and algae wherein said nucleic acid encodes a polypeptide involved in DSMP synthesis. Exemplary sequences are those listed in Tables 1 a, 1 b, 2 and in the section entitled Listing of sequences or sequences with at least 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity thereto. Examples also include sequences which phylogenetically cluster to the said protein families.

[0105] Exemplary enzymes involved in the DMSP synthesis pathways are described in Tables 1 a, 1 b and 2.

[0106] Table 1 a Enzymes involved in the DMSP synthesis pathways- nucleic acid sequences

[0107]

[0108] Table 1 b Enzymes involved in the DMSP synthesis pathways- protein sequences

[0109] For Table 2, see the example section.

[0110] In another aspect, the invention relates to a genetically altered plant wherein said plant expresses a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

[0111] In some embodiments, the genetically altered plant may have any combination of one or more the above exogenous nucleic acid constructs. For example the invention relates to a genetically altered plant wherein said plant expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a SaSDC polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a SaDOX polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto.

[0112] In one embodiment, the exogenous nucleic acid construct comprises a nucleic acid sequence encoding the polypeptides of the invention wherein said nucleic acid sequence is a plant sequence. In one embodiment, the exogenous nucleic acid construct comprises a nucleic acid sequence encoding the polypeptides of the invention wherein said nucleic acid sequence is not a plant sequence.

[0113] In one embodiment, the exogenous nucleic acid construct comprises a nucleic acid sequence encoding the polypeptides of the invention wherein said nucleic acid sequence is eukaryotic, for example an algae sequence. In one embodiment, the exogenous nucleic acid construct comprises a nucleic acid sequence encoding the polypeptides of the invention wherein said nucleic acid sequence is prokaryotic, for example a bacterial sequence.

[0114] In one embodiment, the genetically altered plant produces DMSP from methionine via the methylation, transamination and / or decarboxylation pathway. In one embodiment, the genetically altered plant produces DMSP from methionine via the methylation pathway. In one embodiment, the genetically altered plant produces DMSP from methionine via the transamination pathway. In one embodiment, the genetically altered plant produces DMSP from methionine via the decarboxylation pathway. In one embodiment, the genetically altered plant produces DMSP from methionine via the methylation and the transamination pathway. In one embodiment, the genetically altered plant produces DMSP from methionine via the methylation and the decarboxylation pathway. In one embodiment, the genetically altered plant produces DMSP from methionine via the transamination and the decarboxylation pathway.

[0115] As used herein, the words "nucleic acid", "nucleic acid sequence", "nucleotide", "nucleic acid molecule" or "polynucleotide" are intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), naturally occurring, mutated, synthetic DNA or RNA molecules, and analogs of the DNA or RNA generated using nucleotide analogs. It can be single-stranded or doublestranded. Such nucleic acids or polynucleotides include, but are not limited to, coding sequences of structural genes, anti-sense sequences, and non-coding regulatory sequences that do not encode mRNAs or protein products. These terms also encompass a gene. The term "gene", "allele" or "gene sequence" is used broadly to refer to a DNA nucleic acid associated with a biological function. Thus, genes may include introns and exons as in the genomic sequence, or may comprise only a coding sequence as in cDNAs, and / or may include cDNAs in combination with regulatory sequences. Thus, according to the various aspects of the invention, genomic DNA, cDNA or coding DNA may be used. In one embodiment, the nucleic acid is cDNA or coding DNA.

[0116] The terms "peptide", "polypeptide" and "protein" are used interchangeably herein and refer to amino acids in a polymeric form of any length, linked together by peptide bonds. The term "allele" designates any of one or more alternative forms of a gene at a particular locus. Heterozygous alleles are two different alleles at the same locus. Homozygous alleles are two identical alleles at a particular locus. A wild type (wt) allele is a naturally occurring allele without a modification at the target locus.

[0117] In one embodiment, the term “altering” means “increasing”, “improving” or “enhancing.” Altering the stress response for example can be increased by at least 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%, preferably at least 15% or 20%, more preferably 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% or more in comparison to a control plant, or 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold or 10-fold or more in comparison to a control plant. The stress response can for example be increased by at least 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50%%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% or more in comparison to a control plant or 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9- fold or 10-fold or more in comparison to a control plant.

[0118] The increase in stress response can be measured by measuring yield. The term yield is defined above.

[0119] Plants of the invention have an altered plant stress response compared to a control plant. The term altered plant stress response refers to a stress response that is improved compared to the stress response of a control plant. This is achieved by increased production of DMSP which is protective against environmental stresses. By modulating DMSP levels through transgene expression of genes encoding enzymes in Spartina anglica, namely SaSDC, SaDOX and / or SaMMTI, the inventors demonstrate that DMSP can improve the tolerance of plants to stresses such as salt and drought. The improved stress response protects the plant against detrimental environmental conditions for plant growth and can be quantified for example as a decrease in plant growth rate or as a decrease in biomass accumulation. As explained elsewhere herein, an improved stress response can result in better protection against environmental stresses for example increased drought resistance and ultimately increased yield.

[0120] Spartina anglica synthesises DMSP through the methylation pathway (Figure 18). However, some bacteria also produce DMSP through this methylation pathway via homologous enzymes to SaSDC, SaDOX and SaMMTI. Known homologous enzymes to SaSDC include Burl, homologous enzymes to SaDOX include BurD and homologous enzymes to SaMMTI include MMtN and BurB.

[0121] DMSP is also synthesised by some bacteria, algae and corals through the transamination pathway. Key enzymes include DSYE, DSYB, TpMMT, DsyB, DsyGD. The decarboxylation pathway used by Crypthecodinium cohnii also produces DMSP by converting 3-methylthiopropylamine to 3- methyl mercaptopropionate.

[0122] It is expected that the modulation of DMSP levels through transgene expression of genes encoding enzymes from non-plant sources will improve the stress response of plants upon their expression. In one aspect, the invention also relates to a genetically altered plant wherein said plant expresses one or more exogenous nucleic acid constructs comprising a nucleic acid sequence from plant and / or non-plant sources, such as bacteria, algae and coral. The genetically altered plant may utilise a single, or multiple pathways involved in DMSP synthesis. MMT catalyses the production of SMM from methionine as part of the S-methyl-methionine cycle (SMM cycle) and is the first step of plant DMSP synthesis. The inventors found two transcripts present in S. anglica leaves, SaMMTI and SaMMT2. Only SaMMTI was effective in catalysing the production of SMM. SDC catalyses the production of DMSP-amine from SMM, whilst DOX catalyses the production of DMSP-aldehyde from DMSP-amine. The inventors identified three candidate SDC enzymes predicted to encode an arginine decarboxylase SaADC), ornithine decarboxylase (SaODC) and diaminopimelate decarboxylase SaDAPDC). However, when provided with SMM, only SaODC demonstrated SDC activity. SaODC was termed SaSDC by the inventors. Finally, DMSP- aldehyde dehydrogenase (ALDH) catalyses the production of DMSP from DMSP-aldehyde. The inventors identified two candidate DOX enzymes, SaCAOl and SaCAO2 (termed SaDOX). Surprisingly, SaCAOl was inhibited by endogenous putrescine, therefore was not suitable for use in a transgenic plant. The inventors found that SaMMTI, SaSDC and SaDO work in combination to produce DMSP.

[0123] For the purposes of the invention, a "genetically altered plant" is a plant that has been genetically altered compared to a control plant. A control plant as used according to this aspect of the invention is a plant which has not been modified according to the methods of the invention. Accordingly, the control plant does not express one or more exogenous nucleic acid constructs encoding a SaSDC, SaDOX and / or SaMMTI polypeptide, or other DMSP synthesis enzymes, as described herein. For example, the control plant is a wild type plant. The control plant is typically of the same plant species, preferably the same ecotype or the same or similar genetic background as the plant to be assessed.

[0124] The term "plant" as used in the context of genetically modified plants encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, fruit, shoots, stems, leaves, roots (including tubers), flowers, and tissues and organs, wherein each of the aforementioned comprise the gene / nucleic acid of interest. The term "plant" also encompasses plant cells, suspension cultures, protoplasts, callus tissue, embryos, meristematic regions, gametophytes, sporophytes, pollen and microspores, again wherein each of the aforementioned comprises the gene / nucleic acid of interest.

[0125] The term also extends to harvestable parts of a genetically altered plant of the invention as described above such as, but not limited to seeds, leaves, flowers, stems and roots. The invention furthermore relates to products derived, obtainable or obtained, preferably directly derived, obtainable or obtained, from a harvestable part of such a plant, such as dry pellets or powders, oil, fat and fatty acids, flour, starch or proteins. The invention also relates to food products and food supplements comprising the plant of the invention or parts thereof. In one aspect, the invention relates to a seed of a genetically altered plant of the invention. In another embodiment, the genetically altered plant is a regenerable plant as described herein and cells for use in tissue culture. The tissue culture will preferably be capable of regenerating plants having essentially all of the physiological and morphological characteristics of the plant, and of regenerating plants having substantially the same genotype. Preferably, the regenerable cells in such tissue cultures will be callus, protoplasts, meristematic cells, cotyledons, hypocotyl, leaves, pollen, embryos, roots, root tips, anthers, pistils, shoots, stems, petioles, flowers, and seeds. Still further, the present invention provides plants regenerated from the plant tissue cultures of the invention.

[0126] For the purposes of certain other embodiments of the invention, "transgenic", "transgene" or "recombinant" means with regard to, for example, a nucleic acid sequence, an expression cassette, nucleic acid construct, gene construct or a vector comprising the nucleic acid sequence or an organism transformed with the nucleic acid sequences, expression cassettes or vectors according to the invention, all those constructions brought about by recombinant methods in which either (a) the nucleic acid sequences encoding proteins useful in the methods of the invention, or (b) genetic control sequence(s) which is operably linked with the nucleic acid sequence according to the invention, for example a promoter, or (c) a) and b) are not located in their natural genetic environment or have been modified by recombinant methods.

[0127] The term "functional variant of a nucleic acid sequence" as used herein with reference to SEQ ID NO: 1 , 3, 5, 20, 21 , 22, 23, 24, 25, 26, 27, 28 or a sequence from Table 1 a or 2 refers to a variant gene sequence or part of the gene sequence which retains the biological function of the full nonvariant sequence. A functional variant also comprises a variant of the gene of interest, which has sequence alterations that do not affect function, for example in non-conserved residues. Also encompassed is a variant that is substantially identical, i.e. has only some sequence variations, for example in non-conserved residues, compared to the wild type sequences as shown herein and is biologically active. Alterations in a nucleic acid sequence that results in the production of a different amino acid at a given site that does not affect the functional properties of the encoded polypeptide are well known in the art. For example, a codon for the amino acid alanine, a hydrophobic amino acid, may be substituted by a codon encoding another less hydrophobic residue, such as glycine, or a more hydrophobic residue, such as valine, leucine, or isoleucine. Similarly, changes which result in substitution of one negatively charged residue for another, such as aspartic acid for glutamic acid, or one positively charged residue for another, such as lysine for arginine, can also be expected to produce a functionally equivalent product. Nucleotide changes which result in alteration of the N- terminal and C-terminal portions of the polypeptide molecule would also not be expected to alter the activity of the polypeptide. Each of the proposed modifications is well within the routine skill in the art, as is determination of retention of biological activity of the encoded products.

[0128] The term "functional variant” as used herein with reference to SEQ ID NO: 2, 4, 6, 11 , 12, 13, 31 , 15, 29, 17, 18, 19, 16 or 14 or a sequence from Table 1 b or 2 refers to a variant protein / polypeptide sequence and refers to a variant protein / polypeptide sequence which retains the biological function of the full non-variant sequence.

[0129] The term "functional variant” as used herein with reference to SEQ ID NO: 1 , 3, 5, 20, 21 , 22, 23, 24, 25, 26, 27, 28 or a sequence from Table 1 a or 2 refers to a variant nucleic acid sequence and refers to a variant protein / polypeptide sequence which retains the biological function of the full non-variant sequence.

[0130] In one embodiment, a functional variant has at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the non-variant nucleic acid or polypeptide.

[0131] Two nucleic acid sequences or polypeptides are said to be "identical" if the sequence of nucleotides or amino acid residues, respectively, in the two sequences is the same when aligned for maximum correspondence as described below. The terms "identical" or percent "identity," in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence over a comparison window, as measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection. When percentage of sequence identity is used in reference to proteins or peptides, it is recognised that residue positions that are not identical often differ by conservative amino acid substitutions, where amino acid residues are substituted for other amino acid residues with similar chemical properties (e.g., charge or hydrophobicity) and therefore do not change the functional properties of the molecule. Where sequences differ in conservative substitutions, the percent sequence identity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those of skill in the art. For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters. Nonlimiting examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms.

[0132] Suitable functional variants can be identified by sequence comparisons and identifications of conserved domains. There are predictors in the art that can be used to identify such sequences. The function of the homologue can be identified as described herein and a skilled person would thus be able to confirm the function, for example when overexpressed in a plant.

[0133] Thus, the nucleotide sequences of the invention and described herein can also be used to isolate corresponding sequences from other organisms, particularly other plants, for example crop plants. In this manner, methods such as PCR, hybridization, and the like can be used to identify such sequences based on their sequence homology to the sequences described herein. Topology of the sequences and the characteristic domains structure can also be considered when identifying and isolating homologs. Sequences may be isolated based on their sequence identity to the entire sequence or to fragments thereof. In hybridization techniques, all or part of a known nucleotide sequence is used as a probe that selectively hybridizes to other corresponding nucleotide sequences present in a population of cloned genomic DNA fragments or cDNA fragments (i.e., genomic or cDNA libraries) from a chosen plant. The hybridization probes may be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and may be labelled with a detectable group, or any other detectable marker. Methods for preparation of probes for hybridization and for construction of cDNA and genomic libraries are generally known in the art and are disclosed in Sambrook, et al., (1989) Molecular Cloning: A Library Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, New York).

[0134] Hybridization of such sequences may be carried out under stringent conditions. By "stringent conditions" or "stringent hybridization conditions" is intended conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g. at least 2-fold over background). Stringent conditions are sequence dependent and will be different in different circumstances. By controlling the stringency of the hybridization and / or washing conditions, target sequences that are 100% complementary to the probe can be identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity are detected (heterologous probing). Generally, a probe is less than about 1000 nucleotides in length, preferably less than 500 nucleotides in length.

[0135] Typically, stringent conditions will be those in which the salt concentration is less than about 1.5 M Na+ion, typically about 0.01 to 1 .0 M Na+ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes (e.g., 10 to 50 nucleotides) and at least about 60°C for long probes (e.g., greater than 50 nucleotides). Duration of hybridization is generally less than about 24 hours, usually about 4 to 12. Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide.

[0136] In a further embodiment, a functional variant as used herein comprises a nucleic acid sequence encoding a SaMMTI , SaSDC, SaDOX, Burl, BurD, DsyGD, DSYE, DysB, DYSB or TpMMT polypeptide as defined herein that is capable of hybridising under stringent conditions as defined herein to a nucleic acid sequence as defined in SEQ ID NO: 1 , 3, 5, 20, 21 , 22, 23, 24, 25, 26, 27 or 28 or a sequence as shown in Table 1 a.

[0137] In one embodiment, the genetically altered plant expresses a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and expressing an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto.

[0138] In one embodiment, the genetically altered plant of a. further expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a MmtN polypeptide comprising SEQ ID NO:18 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurB polypeptide comprising SEQ ID NO:19 or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, the genetically altered plant of a. optionally further expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an DMSP-aldehyde dehydrogenase (ALDH) and / or a BurE polypeptide or a functional variant thereof having at least 90% sequence identity thereto or a polypeptide selected from Table 1 b or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, the BurE polypeptide comprises SEQ ID NO: 30 or a functional variant thereof having at least 90% sequence identity thereto.

[0139] In one embodiment, the genetically altered plant expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a SaSDC polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a SaDOX polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto.

[0140] In one embodiment, the genetically altered plant further expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a SaMMTI polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto.

[0141] Thus, in one embodiment, the genetically altered plant expresses an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a SaSDC polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and expressing an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a SaDOX polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a SaMMTI polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto.

[0142] In one embodiment of the genetically altered plant, said nucleic acid sequences form part of one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve nucleic acid constructs. For example, a single nucleic acid construct may be used comprising a nucleic acid sequence encoding a SaSDC and SaDOX polypeptide. For example, a single nucleic acid construct may be used comprising a nucleic acid sequence encoding a SaSDC, a SaDOX, and a SaMMTI polypeptide. For example, a single nucleic acid construct may be used comprising a nucleic acid sequence encoding a. a SaSDC and / or a Burl and a SaDOX and / or a BurD and a SaMMTI and / or MmtN and / or BurB polypeptide; and / or b. a DsyGD and a DSYE and / or a DsyB and / or a DSYB and / or a TpMMT polypeptide.

[0143] Exemplary sequences are in Table 1 , 2 and listed elsewhere herein.

[0144] Alternatively, each nucleic acid sequence may be comprised in a single construct.

[0145] In one embodiment, the construct comprises or consists of a nucleic acid encoding a polypeptide of SEQ ID: 2, 4, 6, 11 , 12, 13, 31 , 15, 29, 17, 18, 19, 16 or 14 or a sequence shown in Table 1 b.

[0146] In one embodiment, the functional variant is a homolog of SEQ ID NO: 2, 4, 6, 11 , 12, 13, 31 , 15, 29, 17, 18, 19, 16 or 14 or a sequence shown in Table 1 b or respectively which is a homolog of a plant that accumulates DMSP. In one embodiment, the functional variant is a homolog of SEQ ID NO: 2, 4, 6, 11 , 12, 13, 31 , 15, 29, 17, 18, 19, 16 or 14 or a sequence shown in Table 1 b respectively which is a homolog of a plant that does not accumulate DMSP, e.g. the plant produces no endogenous DMSP or low levels of endogenous DMSP.

[0147] In one embodiment of the genetically altered plant, the nucleic acid encoding a SaSDC polypeptide comprises SEQ ID NO:1 or a functional variant thereof having at least 90% sequence identity thereto.

[0148] In one embodiment of the genetically altered plant, the nucleic acid encoding a SaDOX polypeptide comprises SEQ ID NO:3 or a functional variant thereof having at least 90% sequence identity thereto.

[0149] In one embodiment of the genetically altered plant, the nucleic acid encoding a SaMMT 1 polypeptide comprises SEQ ID NO:5 or a functional variant thereof having at least 90% sequence identity thereto.

[0150] In one embodiment of the genetically altered plant, the nucleic acid encoding a Burl polypeptide comprises SEQ ID NO: 20 or a functional variant thereof having at least 90% sequence identity thereto.

[0151] In one embodiment of the genetically altered plant, the nucleic acid encoding a BurD polypeptide comprises SEQ ID NO: 21 or a functional variant thereof having at least 90% sequence identity thereto.

[0152] In one embodiment of the genetically altered plant, the nucleic acid encoding a DsyGD polypeptide comprises SEQ ID NO: 22 or a functional variant thereof having at least 90% sequence identity thereto.

[0153] In one embodiment of the genetically altered plant, the nucleic acid encoding a DSYE polypeptide comprises SEQ ID NO: 23 or a functional variant thereof having at least 90% sequence identity thereto.

[0154] In one embodiment of the genetically altered plant, the nucleic acid encoding a DsyB polypeptide comprises SEQ ID NO: 24 or a functional variant thereof having at least 90% sequence identity thereto.

[0155] In one embodiment of the genetically altered plant, the nucleic acid encoding a DSYB polypeptide comprises SEQ ID NO: 25 or a functional variant thereof having at least 90% sequence identity thereto.

[0156] In one embodiment of the genetically altered plant, the nucleic acid encoding a TpMMT polypeptide comprises SEQ ID NO: 26 or a functional variant thereof having at least 90% sequence identity thereto.

[0157] In one embodiment of the genetically altered plant, the nucleic acid encoding a MmtN polypeptide comprises SEQ ID NO: 27 or a functional variant thereof having at least 90% sequence identity thereto.

[0158] In one embodiment of the genetically altered plant, the nucleic acid encoding a BurB polypeptide comprises SEQ ID NO: 28 or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, the nucleic acid sequence comprises or consists of SEQ ID NO: 1 , 3, 5, 20, 21 , 22, 23, 24, 25, 26, 27, 28 or a sequence as shown in Table 1 a or a functional variant thereof. This encodes a protein comprising or consisting of SEQ ID NO: 2, 4, 6, 11 , 12, 13, 31 , 15, 29, 17, 18, 19 or a sequence as shown in Table 1 b respectively. In one embodiment, the protein comprises or consists of SEQ ID NO: 16 or 14.

[0159] In one embodiment of the genetically altered plant, said plant overexpresses the SaSDC, SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence.

[0160] In one embodiment of the genetically altered plant, the nucleic acid construct comprising a nucleic acid sequence encoding a SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB polypeptide and further comprises a constitutive promoter.

[0161] A "constitutive promoter" refers to a promoter that is transcriptionally active during most, but not necessarily all, phases of growth and development and under most environmental conditions, in at least one cell, tissue or organ. Examples of constitutive promoters include but are not limited to actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promoter, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfa H3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V- ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters. In another aspect of the invention there is provided a vector comprising the nucleic acid sequence described above.

[0162] In one embodiment, said promoter is a plant promoter. As used herein, a "plant promoter" comprises regulatory elements, which mediate the expression of a coding sequence segment in plant cells. Accordingly, a plant promoter need not be of plant origin, but may originate from viruses or microorganisms, for example from viruses which attack plant cells. The "plant promoter" can also originate from a plant cell, e.g. from the plant which is transformed with the nucleic acid sequence to be expressed in the inventive process and described herein. This also applies to other "plant" regulatory signals, such as "plant" terminators. The promoters upstream of the nucleotide sequences useful in the methods of the present invention can be modified by one or more nucleotide substitution(s), insertion(s) and / or deletion(s) without interfering with the functionality or activity of either the promoters, the open reading frame (ORF) or the 3'-regulatory region such as terminators or other 3' regulatory regions which are located away from the ORF. It is furthermore possible that the activity of the promoters is increased by modification of their sequence, or that they are replaced completely by more active promoters, even promoters from heterologous organisms. For expression in plants, the nucleic acid molecule must, as described above, be linked operably to or comprise a suitable promoter which expresses the gene at the right point in time and with the required spatial expression pattern.

[0163] In one embodiment, the promoter is selected from actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promotor, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfa H3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V-ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters.

[0164] Increased expression, as used herein, refers to increase in expression level of at least 2%, or at least 5%, or at least 10%, or at least 15%, or at least 20%, or at least 25%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, 95% or 100% or 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold or 10-fold even more. Said increase is an increase with respect to the expression in control plants.

[0165] As the skilled person will be well aware, various promoters may be used to promote the transcription of the nucleic acid. Such promoters include for example constitutive promoters, inducible promoters (e.g. stress-inducible promoters, drought-inducible promoters, hormone-inducible promoters, chemical-inducible promoters, etc.), tissue-specific promoters, developmental regulated promoters and the like.

[0166] A variety of plant gene promoters that regulate gene expression in response to environmental, hormonal, chemical, developmental signals, and in a tissue-active manner can be used for expression of a sequence in plants. Choice of a promoter is based largely on the phenotype of interest and is determined by such factors as tissue (e.g., seed, fruit, root, pollen, vascular tissue, flower, carpel, etc.), inducibility (e.g., in response to wounding, heat, cold, drought, light, pathogens, etc.), timing, developmental stage, and the like.

[0167] Additional promoters that can be used to practice this invention are those that elicit expression in response to stresses, such as the RD29 promoters that are activated in response to drought, low temperature, salt stress, or exposure to ABA, but also promoters that are induced in response to heat, light (e.g., the pea rbcS-3A promoter, and the maize rbcS promoter, wounding (e.g., wunl), pathogens (such as the PR-I promoter and the PDF 1.2 promoter), and chemicals such as methyl jasmonate or salicylic acid. In addition, the timing of the expression can be controlled by using promoters such as those acting at senescence; or late seed development.

[0168] Another example are salt-inducible promoters such as the salt-inducible NHX1 promoter of rice landrace Pokkali (PKN), the salt inducible promoter of the vacuolar pyrophosphatase from Thellungiella halophila (TsVP1) or the salt-inducible promoter of the Citrus sinensis gene encoding phospholipid hydroperoxide isoform gpxl.

[0169] In some embodiments, tissue-specific and / or developmental stage-specific promoters are used, e.g., promoter that can promote transcription only within a certain time frame of developmental stage within that tissue.

[0170] In some embodiments, nucleic acids used to practice the invention can also be operably linked to plant promoters which are inducible upon exposure to chemicals reagents which can be applied to the plant, such as herbicides or antibiotics. In some embodiments, a promoter may be used whose host range is limited to target plant species, such as corn, rice, barley, wheat, potato or other crops, inducible at any stage of development of the crop.

[0171] In some embodiments, a tissue-specific plant promoter may drive expression of operably linked sequences in specific target tissues. In alternative embodiments, a tissue-specific promoter that drives expression preferentially in the target tissue or cell type, but may also lead to some expression in other tissues as well, is used. The nucleic acid construct may also comprise regulatory sequences as described above. Other regulatory regions include those that mediate targeted integration into the genome using genome editing as further described below. For example, the construct intended for the delivery of the transgene to the target region using the CRISPR genomic editing technology may include flanking DNA regions homologous to the insertion site

[0172] The nucleic acid constructs as described herein may be comprised in a vector.

[0173] The term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked; a plasmid is a species of the genus encompassed by "vector". The term "vector" typically refers to a nucleic acid sequence containing an origin of replication and other entities necessary for replication and / or maintenance in a host cell. Vectors capable of directing the expression of genes and / or nucleic acid sequence to which they are operatively linked are referred to herein as "expression vectors". In general, expression vectors of utility are often in the form of "plasmids" which refer to circular double stranded DNA loops which, in their vector form are not bound to the chromosome, and typically comprise entities for stable or transient expression of the encoded DNA. Other expression vectors can be used in the methods as disclosed herein for example, but are not limited to, plasmids, episomes, bacterial artificial chromosomes, yeast artificial chromosomes, bacteriophages or viral vectors, and such vectors can integrate into the host's genome or replicate autonomously in the particular cell. A vector can be a DNA or RNA vector. Other forms of expression vectors known by those skilled in the art which serve the equivalent functions can also be used, for example self-replicating extrachromosomal vectors or vectors which integrate into a host genome. Preferred vectors are those capable of autonomous replication and / or expression of nucleic acids to which they are linked. Vectors capable of directing the expression of genes to which they are operatively linked are referred to herein as "expression vectors".

[0174] The term "regulatory sequences" is used interchangeably with "regulatory elements" herein refers to a segment of nucleic acid, typically but not limited to DNA or RNA or analogues thereof, that modulates the transcription of the nucleic acid sequence to which it is operatively linked, and thus act as transcriptional modulators. Regulatory sequences modulate the expression of gene and / or nucleic acid sequences to which they are operatively linked. Regulatory sequences often comprise "regulatory elements" which are nucleic acid sequences that are transcription binding domains and are recognized by the nucleic acid-binding domains of transcriptional proteins and / or transcription factors, repressors or enhancers etc. Typical regulatory sequences include, but are not limited to, transcriptional promoters, inducible promoters and transcriptional elements, an optional operate sequence to control transcription, a sequence encoding suitable mRNA ribosomal binding sites, and sequences to control the termination of transcription and / or translation. Regulatory sequences can be a single regulatory sequence or multiple regulatory sequences or modified regulatory sequences or fragments thereof. Modified regulatory sequences are regulatory sequences where the nucleic acid sequence has been changed or modified by some means, for example, but not limited to, mutation, methylation etc.

[0175] The term "operatively linked" as used herein refers to the functional relationship of the nucleic acid sequences with regulatory sequences of nucleotides, such as promoters, enhancers, transcriptional and translational stop sites, and other signal sequences. For example, operative linkage of nucleic acid sequences, typically DNA, to a regulatory sequence or promoter region refers to the physical and functional relationship between the DNA and the regulatory sequence or promoter such that the transcription of such DNA is initiated from the regulatory sequence or promoter, by an RNA polymerase that specifically recognizes, binds and transcribes the DNA. In order to optimize expression and / or in vitro transcription, it may be necessary to modify the regulatory sequence for the expression of the nucleic acid or DNA in the cell type for which it is expressed. The desirability of, or need of, such modification may be empirically determined. Enhancers need not be located in close proximity to the coding sequences whose transcription they enhance. Furthermore, a gene transcribed from a promoter regulated in trans by a factor transcribed by a second promoter may be said to be operatively linked to the second promoter. In such a case, transcription of the first gene is said to be operatively linked to the first promoter and is also said to be operatively linked to the second promoter.

[0176] In one embodiment, the exogenous nucleic acid construct is stably incorporated into the plant genome.

[0177] Recently, genome editing techniques have emerged as alternative methods to conventional mutagenesis methods (such as physical and chemical mutagenesis) or methods using the expression of transgenes in plants to produce mutant plants with improved phenotypes that are important in agriculture. These techniques employ sequence-specific nucleases (SSNs) including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the RNA- guided nucleases (e.g. Cas9), which generate targeted DNA double-strand breaks (DSBs), which are then repaired mainly by either error-prone non-homologous end joining (NHEJ) or high-fidelity homologous recombination (HR). Thus, nucleic acid constructs according to the invention can be introduced into plants using targeted genome modification based on such editing techniques.

[0178] Targeted DNA integration into known locations in the genome has potential advantages over the random insertional events typically achieved using conventional means of genetic modification. This avoids screening of a large number of transgenic lines and directly obtain highly productive lines producing recombinant proteins with target genes delivered to transcriptionally active regions. Knock-in lines where the exogenous nucleic acid integrates into the target genome can be achieved in this way allowing integration of transgenic DNA into the plant genome, for example in regions that have a very high constitutive level of expression to achieve overexpression, such as the region of the histone H3.3 gene.

[0179] Thus, in one embodiment, the exogenous nucleic acid construct having a nucleic acid sequences as described herein is inserted into the plant genome using targeted genome modification.

[0180] For example, the nucleic acid construct is inserted into the plant genome using a rare-cutting endonuclease, for example a TALEN, ZFN or CRISPR / Cas9.

[0181] Targeted genome modification or targeted genome editing is a genome engineering technique that uses targeted DNA double-strand breaks (DSBs) to stimulate genome editing through homologous recombination (HR)-mediated recombination events. Site-specific gene integration via CRISPR / Cas9 relies on endogenous DNA repair machinery, which is used to repair genomic DNA damage due to DSBs created by Cas9 or equivalent enzymes.

[0182] To achieve effective genome editing via introduction of site-specific DNA DSBs, four major classes of customizable DNA binding proteins can be used: meganucleases derived from microbial mobile genetic elements, ZF nucleases based on eukaryotic transcription factors, rare-cutting endonucleases / sequence specific endonucleases (SSN), for example TALENs, transcription activator-like effectors (TALEs) from Xanthomonas bacteria, and the RNA-guided DNA endonuclease Cas9 from the type II bacterial adaptive immune system CRISPR (clustered regularly interspaced short palindromic repeats). Meganuclease, ZF, and TALE proteins all recognize specific DNA sequences through protein-DNA interactions. Although meganucleases integrate their nuclease and DNA-binding domains, ZF and TALE proteins consist of individual modules targeting 3 or 1 nucleotides (nt) of DNA, respectively. ZFs and TALEs can be assembled in desired combinations and attached to the nuclease domain of Fokl to direct nucleolytic activity toward specific genomic loci.

[0183] Upon delivery into host cells via the bacterial type III secretion system, TAL effectors enter the nucleus, bind to effector-specific sequences in host gene promoters and activate transcription. Their targeting specificity is determined by a central domain of tandem, 33-35 amino acid repeats. This is followed by a single truncated repeat of 20 amino acids. The majority of naturally occurring TAL effectors examined have between 12 and 27 full repeats.

[0184] These repeats only differ from each other by two adjacent amino acids, their repeat- variable diresidue (RVD). The RVD determines which single nucleotide the TAL effector will recognize: one RVD corresponds to one nucleotide, with the four most common RVDs each preferentially associating with one of the four bases. Naturally occurring recognition sites are uniformly preceded by a T that is required for TAL effector activity. TAL effectors can be fused to the catalytic domain of the Fokl nuclease to create a TAL effector nuclease (TALEN) which makes targeted DNA double- strand breaks (DSBs) in vivo for genome editing. The use of this technology in genome editing is well described in the art, for example in US 8,440,431 , US 8,440, 432 and US 8,450,471. Customized plasmids can be used with the Golden Gate cloning method to assemble multiple DNA fragments. The Golden Gate method uses Type IIS restriction endonucleases, which cleave outside their recognition sites to create unique 4 bp overhangs. Cloning is expedited by digesting and ligating in the same reaction mixture because correct assembly eliminates the enzyme recognition site. Assembly of a custom TALEN or TAL effector construct and involves two steps: (i) assembly of repeat modules into intermediary arrays of 1-10 repeats and (ii) joining of the intermediary arrays into a backbone to make the final construct.

[0185] Another genome editing method that can be used according to the various aspects of the invention is CRISPR. The use of this technology in genome editing is well described in the art, for example in US 8,697,359. In short, CRISPR is a microbial nuclease system involved in defence against invading phages and plasmids. CRISPR loci in microbial hosts contain a combination of CRISPR- associated (Cas) genes as well as non-coding RNA elements capable of programming the specificity of the CRISPR-mediated nucleic acid cleavage. Three types (l-lll) of CRISPR systems have been identified across a wide range of bacterial hosts. One key feature of each CRISPR locus is the presence of an array of repetitive sequences (direct repeats) interspaced by short stretches of non-repetitive sequences (spacers). The non-coding CRISPR array is transcribed and cleaved within direct repeats into short crRNAs containing individual spacer sequences, which direct Cas nucleases to the target site (protospacer).

[0186] The Type II CRISPR is one of the most well characterized systems and carries out targeted DNA double-strand breaks in four sequential steps. First, two non-coding RNA, the pre-crRNA array and tracrRNA, are transcribed from the CRISPR locus. Second, tracrRNA hybridizes to the repeat regions of the pre-crRNA and mediates the processing of pre-crRNA into mature crRNAs containing individual spacer sequences. Third, the mature crRNA: tracrRNA complex directs Cas9 to the target DNA via Watson-Crick base-pairing between the spacer on the crRNA and the protospacer on the target DNA next to the protospacer adjacent motif (PAM), an additional requirement for target recognition. Finally, the Cas enzyme, for example Cas9 mediates cleavage of target DNA to create a double-stranded break within the protospacer. Cas9 is thus the hallmark protein of the type II CRISPR-Cas system, and a large monomeric DNA nuclease guided to a DNA target sequence adjacent to the PAM sequence motif by a complex of two noncoding RNAs: CRIPSR RNA (crRNA) and trans-activating crRNA (tracrRNA). The Cas9 protein contains two nuclease domains homologous to RuvC and HNH nucleases. The HNH nuclease domain cleaves the complementary DNA strand whereas the RuvC-like domain cleaves the non-complementary strand and, as a result, a blunt cut is introduced in the target DNA. Heterologous expression of Cas9 together with a guide RNA (gRNA) also called single guide RNA (sgRNA) can introduce site-specific double strand breaks (DSBs) into genomic DNA of live cells from various organisms. For applications in eukaryotic organisms, codon optimized versions of Cas9, which is originally from the bacterium Streptococcus pyogenes, have been used.

[0187] Synthetic CRISPR systems typically consist of two components, the gRNA and a non-specific CRISPR-associated endonuclease and can be used to generate knock-out cells or animals by coexpressing a gRNA specific to the gene to be targeted and capable of association with the endonuclease, for example Cas9. Notably, the gRNA is an artificial molecule comprising one domain interacting with the Cas or any other CRISPR effector protein or a variant or catalytically active fragment thereof and another domain interacting with the target nucleic acid of interest and thus representing a synthetic fusion of crRNA and tracrRNA. The genomic target can be any 20 nucleotide DNA sequence, provided that the target is present immediately upstream of a PAM sequence. The PAM sequence is of outstanding importance for target binding and the exact sequence is dependent upon the species of Cas enzyme, for example Cas9.

[0188] In one embodiment, the Cas endonuclease gene is a Cas9 endonuclease, such as but not limited to, Cas9 genes listed in W02007 / 025097 incorporated herein by reference. In another embodiment, the Cas endonuclease gene is plant, maize or soybean optimized Cas9 endonuclease. In one embodiment, the Cas endonuclease gene is an alternate DNA-targeting Cas endonuclease such as Cas12.

[0189] In one embodiment, the Cas endonuclease gene is a plant codon optimized streptococcus pyogenes Cas9 gene that can recognize any genomic sequence of the form N(12-30)NGG can in principle be targeted.

[0190] In a specific embodiment Cas9 is codon-optimised Cas9. In one embodiment the alternate DNA- targeting Cas endonuclease is codon-optimised. For example, the alternate DNA-targeting Cas endonuclease is codon-optimised Cas12. In another embodiment, the CRISPR enzyme is a protein from the family of Class 2 candidate proteins, such as C2c1 , C2C2 and / or C2c3. In one embodiment, the Cas protein is from Streptococcus pyogenes. In an alternative embodiment, the Cas protein may be from any one of Staphylococcus aureus, Neisseria meningitides or Streptococcus thermophiles.

[0191] In a further embodiment, the Cas9 or the alternate DNA-targeting Cas endonuclease protein has been modified to improve activity. Suitable homologs or orthologs can be identified by sequence comparisons and identifications of conserved domains. The function of the homolog or ortholog can be identified as described herein and a skilled person would thus be able to confirm the function when expressed in a plant. In a further embodiment, the Cas9 protein or alternate DNA-targeting Cas endonuclease has been modified to improve activity. For example, in one embodiment, the Cas9 protein may comprise the D10A amino acid substitution, this nickase cleaves only the DNA strand that is complementary to and recognized by the gRNA. In an alternative embodiment, the Cas9 protein may alternatively or additionally comprise the H840A amino acid substitution, this nickase cleaves only the DNA strand that does not interact with the sRNA. In this embodiment, Cas9 may be used with a pair (i.e. two) sgRNA molecules (or a construct expressing such a pair) and as a result can cleave the target region on the opposite DNA strand, with the possibility of improving specificity by 100-1500 fold. In a further embodiment, the Cas9 protein may comprise a D1135E substitution. The Cas9 protein may also be the VQR variant. Alternatively, the Cas protein may comprise a mutation in both nuclease domains, HNH and RuvC-like and therefore is catalytically inactive. Rather than cleaving the target strand, this catalytically inactive Cas protein can be used to prevent the transcription elongation process, leading to a loss of function of incompletely translated proteins when co-expressed with a sgRNA molecule. An example of a catalytically inactive protein is dead Cas9 (dCas9) caused by a point mutation in RuvC and / or the HNH nuclease domains.

[0192] Suitable methods for producing the CRISPR nucleic acids and vectors system are known, and for example are published in Molecular Plant (Ma et al., 2015, Molecular Plant, 2015 Aug;8(8):1274-8), which is incorporated herein by reference.

[0193] Most of the genetic constructs intended for the delivery of transgenes to the target region using the CRISPR / Cas genomic editing technology include flanking DNA regions homologous to the insertion site. To increase the efficiency of the delivery of the target construct to a given region of the genome a technique can be used that allows the release of the target fragment in the cell in the form of a linear template from the circular plasmid DNA with the help of the Cas nuclease (Permyakova et al, Int J Mol Sci. 2022 Aug; 23(15)).

[0194] Alternatively, conventional methods for integrating the exogenous nucleic acid construct comprising the transgene can be used, e.g. transformation.

[0195] The transfer of foreign genes into the genome of a plant is called transformation. Transformation of plants is now a routine technique in many species. Advantageously, any of several transformation methods may be used to introduce the gene of interest into a suitable ancestor cell. The methods described for the transformation and regeneration of plants from plant tissues or plant cells may be utilized for transient or for stable transformation. Transformation methods include the use of liposomes, electroporation, chemicals that increase free DNA uptake, injection of the DNA directly into the plant, particle bombardment as described in the examples, transformation using viruses or pollen and microinjection. Methods may be selected from the calcium / polyethylene glycol method for protoplasts, electroporation of protoplasts, microinjection into plant material, DNA or RNA-coated particle bombardment, infection with (non-integrative) viruses and the like. Transgenic plants, including transgenic crop plants, are for example produced via Agrobacterium tumefaciens mediated transformation.

[0196] The details of this method are well known to a skilled person. In short, plant transformation by Agrobacterium results in the integration into the nuclear genome of a sequence called T-DNA, which is carried on a bacterial plasmid. The use of T-DNA transformation leads to stable single insertions. Further analysis of the resultant transformed lines is straightforward and each individual insertion line can be rapidly characterized by direct sequencing and analysis of DNA flanking the insertion. Gene expression in the mutant is compared to expression of the SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence in a control plant, e.g. wild type plant and phenotypic analysis is also carried out.

[0197] To select transformed plants, the plant material obtained in the transformation is, as a rule, subjected to selective conditions so that transformed plants can be distinguished from untransformed plants. For example, the seeds obtained in the above-described manner can be planted and, after an initial growing period, subjected to a suitable selection by spraying. A further possibility is growing the seeds, if appropriate after sterilization, on agar plates using a suitable selection agent so that only the transformed seeds can grow into plants. Alternatively, the transformed plants are screened for the presence of a selectable marker.

[0198] Following DNA transfer and regeneration, putatively transformed plants may also be evaluated, for instance using Southern analysis, for the presence of the gene of interest, copy number and / or genomic organisation. Alternatively or additionally, expression levels of the newly introduced DNA may be monitored using Northern and / or Western analysis, both techniques being well known to persons having ordinary skill in the art.

[0199] The generated transformed plants may be propagated by a variety of means, such as by clonal propagation or classical breeding techniques. For example, a first generation (or T1) transformed plant may be selfed and homozygous second-generation (or T2) transformants selected, and the T2 plants may then further be propagated through classical breeding techniques.

[0200] The term "introduction" or "transformation" as referred to herein encompasses the transfer of an exogenous polynucleotide into a host cell, irrespective of the method used for transfer. Plant tissue capable of subsequent clonal propagation, whether by organogenesis or embryogenesis, may be transformed with a genetic construct of the present invention and a whole plant regenerated there from. The particular tissue chosen will vary depending on the clonal propagation systems available for, and best suited to, the particular species being transformed. Exemplary tissue targets include leaf disks, pollen, embryos, cotyledons, hypocotyls, megagametophytes, callus tissue, existing meristematic tissue (e.g., apical meristem, axillary buds, and root meristems), and induced meristem tissue (e.g., cotyledon meristem and hypocotyl meristem). The resulting transformed plant cell may then be used to regenerate a transformed plant in a manner known to persons skilled in the art.

[0201] In one embodiment, said plant is a dicot or monocot plant. These terms are defined above.

[0202] The stress is abiotic or biotic stress as further defined above.

[0203] Nucleic acids, polypeptides, vectors and host cells

[0204] In another aspect, the invention relates to an isolated polypeptide comprising or consisting of SEQ ID NO: 2 or a sequence having 90% sequence identity thereto. In one embodiment, sequence identity is 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. In another aspect, the invention relates to an isolated polypeptide comprising or consisting of SEQ ID NO: 4 or a sequence having 92% sequence identity thereto. In one embodiment, sequence identity is 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0205] In another aspect, the invention relates to an isolated polypeptide comprising or consisting of SEQ ID NO: 6 or a sequence having 90% sequence identity thereto. In one embodiment, sequence identity is 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0206] In another aspect, the invention relates to an isolated nucleic acid sequence comprising or consisting of SEQ ID NO: 1 or a sequence having 90% sequence identity thereto. In one embodiment, sequence identity is 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0207] In another aspect, the invention relates to an isolated nucleic acid sequence comprising or consisting of SEQ ID NO: 3 or a sequence having 92% sequence identity thereto. In one embodiment, sequence identity is 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0208] In another aspect, the invention relates to an isolated nucleic acid sequence comprising or consisting of SEQ ID NO: 5 or a sequence having 90% sequence identity thereto. In one embodiment, sequence identity is 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0209] In another aspect, the invention relates to a vector comprising an isolated nucleic acid sequence as above or a sequence as shown in Table 1 a or a sequence having 90% sequence identity thereto. In one embodiment, sequence identity is 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0210] In one embodiment, the vector further comprises a promoter sequence operably linked to said nucleic acid sequence.

[0211] In another aspect, the invention relates to a host cell comprising an isolated nucleic acid sequence or the vector as described above. The host cell may be a plant, viral or bacterial cell. The host cell may be a plant cell or a microbial cell. The host cell may be a bacterial cell, such as Agrobacterium tumefaciens, or an isolated plant cell. The invention also relates to a culture medium or kit comprising a culture medium and an isolated host cell.

[0212] In a further aspect of the invention, there is provided an isolated plant cell transfected with at least one nucleic acid construct as described herein. In one embodiment, the isolated plant cell is transfected with at least one nucleic acid construct as described herein and a second nucleic acid construct, wherein said second nucleic acid construct comprises a nucleic acid sequence encoding a Cas protein, preferably a Cas9 protein or a functional variant thereof. Preferably, the second nucleic acid construct is transfected before, after or concurrently with the first nucleic acid construct described herein. In an alternative aspect of the invention, the nucleic acid construct comprises at least one nucleic acid sequence that encodes a TAL effector.

[0213] The invention also relates to a nucleic acid construct comprising a nucleic acid sequence operably linked to a suitable plant promoter. A suitable plant promoter may be a constitutive or strong promoter as described herein or may be a tissue-specific promoter. In one embodiment, suitable plant promoters are selected from, but not limited to, oestrum yellow leaf curling virus (CmYLCV) promoter, switchgrass ubiquitin 1 promoter (PvUbil), wheat U6 RNA polymerase III (TaU6), CaMV35S, wheat U6 or maize ubiquitin (e.g. Ubi 1) promoters. Alternatively, expression can be specifically directed to particular tissues of through gene expression-regulating sequences.

[0214] In another aspect, the invention relates to a use of a nucleic acid sequence or vector as described above for altering the stress response of a plant.

[0215] Method for altering the stress response

[0216] In another aspect, the invention relates to a method for altering the stress response of a plant, comprising introducing into said plant a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

[0217] In one embodiment, the method comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S-methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto.

[0218] In one embodiment, the method comprises introducing into said plant a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, a. further comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a MmtN polypeptide comprising SEQ ID NO:18 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurB polypeptide comprising SEQ ID NO:19 or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, the genetically altered plant of a. optionally further expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an DMSP-aldehyde dehydrogenase (ALDH) and / or a BurE polypeptide or a polypeptide selected from Table 1 b or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, the BurE polypeptide comprises SEQ ID NO: 30 or a functional variant thereof having at least 90% sequence identity thereto.

[0219] In one embodiment, the method comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S-methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto.

[0220] In one embodiment, the method comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto.

[0221] In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is a plant sequence. In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is not a plant sequence.

[0222] In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is eukaryotic, for example an algae sequence.

[0223] In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is prokaryotic, for example a bacterial sequence.

[0224] In one embodiment, the genetically altered plant produces DMSP from methionine via the methylation, transamination and / or decarboxylation pathway.

[0225] In one embodiment, said nucleic acid sequences form part of one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve nucleic acid constructs. In one embodiment, the nucleic acid encoding a SaSDC polypeptide comprises SEQ ID NO:1 or a functional variant thereof having at least 90% sequence identity thereto.

[0226] In one embodiment, the nucleic acid encoding a SaDOX polypeptide comprises SEQ ID NO:3 or a functional variant thereof having at least 90% sequence identity thereto.

[0227] In one embodiment, the nucleic acid encoding a SaMMTI polypeptide comprises SEQ ID NO:5 or a functional variant thereof having at least 90% sequence identity thereto.

[0228] In one embodiment, the nucleic acid encoding a Burl polypeptide comprises SEQ ID NO: 20 or a functional variant thereof having at least 90% sequence identity thereto.

[0229] In one embodiment, the nucleic acid encoding a BurD polypeptide comprises SEQ ID NO: 21 or a functional variant thereof having at least 90% sequence identity thereto.

[0230] In one embodiment, the nucleic acid encoding a DsyGD polypeptide comprises SEQ ID NO: 22 or a functional variant thereof having at least 90% sequence identity thereto.

[0231] In one embodiment, the nucleic acid encoding a DSYE polypeptide comprises SEQ ID NO: 23 or a functional variant thereof having at least 90% sequence identity thereto.

[0232] In one embodiment, the nucleic acid encoding a DsyB polypeptide comprises SEQ ID NO: 24 or a functional variant thereof having at least 90% sequence identity thereto.

[0233] In one embodiment, the nucleic acid encoding a DSYB polypeptide comprises SEQ ID NO: 25 or a functional variant thereof having at least 90% sequence identity thereto.

[0234] In one embodiment, the nucleic acid encoding a TpMMT polypeptide comprises SEQ ID NO: 26 or a functional variant thereof having at least 90% sequence identity thereto.

[0235] In one embodiment, the nucleic acid encoding a MmtN polypeptide comprises SEQ ID NO: 27 or a functional variant thereof having at least 90% sequence identity thereto.

[0236] In one embodiment, the nucleic acid encoding a BurB polypeptide comprises SEQ ID NO: 28 or a functional variant thereof having at least 90% sequence identity thereto.

[0237] In one embodiment, the nucleic acid encodes a polypeptide comprising a sequence as shown in Table 1 b or a functional variant thereof.

[0238] In one embodiment, said plant expresses, for example overexpresses an exogenous SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence. Exemplary sequences are shown herein.

[0239] In one embodiment, the nucleic acid construct comprises a nucleic acid sequence encoding a SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB polypeptide further comprises a constitutive promoter.

[0240] In one embodiment, said promoter is selected from actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promoter, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfa H3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V-ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters.

[0241] In one embodiment, the exogenous nucleic acid construct is stably incorporated into the plant genome.

[0242] In one embodiment, the exogenous nucleic acid construct is inserted into the plant genome using targeted genome modification.

[0243] In one embodiment, the nucleic acid construct is inserted into the plant genome using a rare-cutting endonuclease, for example a TALEN, ZFN or CRISPR / Cas9. Further details are set out above.

[0244] In another embodiment, the nucleic acid construct is introduced using transformation. Further details are set out above.

[0245] In one embodiment, said plant is a dicot or monocot plant.

[0246] In one embodiment, said dicot or monocot plant is selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco, grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut, barley or oats. In one embodiment the plant may be selected from a plant as described above.

[0247] In one embodiment, the stress is abiotic or biotic stress.

[0248] In one embodiment, the abiotic stress is selected from osmotic stress, oxidative stress, heavy metal stress, drought, submergence, cold, UV radiation, heat, ozone, nutrient deficiency and / or salt stress.

[0249] Methods for producing a plant

[0250] In another aspect, the invention relates to a method for producing a plant with an altered stress response, comprising introducing into said plant a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

[0251] In one embodiment, the method comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S-methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto.

[0252] In one embodiment, the method comprises introducing into said plant a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and expressing an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto.

[0253] In one embodiment, a. further comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S-methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto. In one embodiment, the genetically altered plant of a. optionally further expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an DMSP-aldehyde dehydrogenase (ALDH) and / or a BurE polypeptide. In one embodiment, the BurE polypeptide comprises SEQ ID NO: 30 or a functional variant thereof having at least 90% sequence identity thereto.

[0254] In one embodiment, the method comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto.

[0255] In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is a plant sequence. In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is not a plant sequence.

[0256] In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is eukaryotic, for example an algae sequence.

[0257] In one embodiment, the nucleic acid sequence in the exogenous nucleic acid construct encoding the polypeptides of the invention is prokaryotic, for example bacterial sequence.

[0258] In one embodiment, the genetically altered plant produces DMSP from methionine via the methylation, transamination and / or decarboxylation pathway.

[0259] In one embodiment, said nucleic acid sequences form part of one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve nucleic acid constructs. In one embodiment, the nucleic acid encoding a SaSDC polypeptide comprises SEQ ID NO:1 or a functional variant thereof having at least 90% sequence identity thereto.

[0260] In one embodiment, the nucleic acid encoding a SaDOX polypeptide comprises SEQ ID NO:3 or a functional variant thereof having at least 90% sequence identity thereto.

[0261] In one embodiment, the nucleic acid encoding a SaMMTI polypeptide comprises SEQ ID NO:5 or a functional variant thereof having at least 90% sequence identity thereto.

[0262] In one embodiment, the nucleic acid encoding a Burl polypeptide comprises SEQ ID NO: 20 or a functional variant thereof having at least 90% sequence identity thereto.

[0263] In one embodiment, the nucleic acid encoding a BurD polypeptide comprises SEQ ID NO: 21 or a functional variant thereof having at least 90% sequence identity thereto.

[0264] In one embodiment, the nucleic acid encoding a DsyGD polypeptide comprises SEQ ID NO: 22 or a functional variant thereof having at least 90% sequence identity thereto.

[0265] In one embodiment, the nucleic acid encoding a DSYE polypeptide comprises SEQ ID NO: 23 or a functional variant thereof having at least 90% sequence identity thereto.

[0266] In one embodiment, the nucleic acid encoding a DsyB polypeptide comprises SEQ ID NO: 24 or a functional variant thereof having at least 90% sequence identity thereto.

[0267] In one embodiment, the nucleic acid encoding a DSYB polypeptide comprises SEQ ID NO: 25 or a functional variant thereof having at least 90% sequence identity thereto.

[0268] In one embodiment, the nucleic acid encoding a TpMMT polypeptide comprises SEQ ID NO: 26 or a functional variant thereof having at least 90% sequence identity thereto.

[0269] In one embodiment, the nucleic acid encoding a MmtN polypeptide comprises SEQ ID NO: 27 or a functional variant thereof having at least 90% sequence identity thereto.

[0270] In one embodiment, the nucleic acid encoding a BurB polypeptide comprises SEQ ID NO: 28 or a functional variant thereof having at least 90% sequence identity thereto.

[0271] In one embodiment, the nucleic acid encodes a polypeptide as shown in Tablel b or a functional variant thereof having at least 90% sequence identity thereto.

[0272] In one embodiment, said plant expresses, for example overexpresses an exogenous SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence. Exemplary sequences are shown herein.

[0273] In one embodiment, the nucleic acid construct comprises a nucleic acid sequence encoding a SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB polypeptide further comprises a constitutive promoter.

[0274] In one embodiment, said promoter is selected from actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promoter, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfa H3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V-ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters.

[0275] In one embodiment, the exogenous nucleic acid construct is stably incorporated into the plant genome.

[0276] In one embodiment, the exogenous nucleic acid construct is inserted into the plant genome using targeted genome modification.

[0277] In one embodiment, the nucleic acid construct is inserted into the plant genome using a rare-cutting endonuclease, for example a TALEN, ZFN or CRISPR / Cas9. Further details are set out above.

[0278] In another embodiment, the nucleic acid construct is introduced using transformation. Further details are set out above.

[0279] In one embodiment, said plant is a dicot or monocot plant.

[0280] In one embodiment, said dicot or monocot plant is selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco, grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut, barley or oats. In one embodiment the plant may be selected from a plant as described above.

[0281] In one embodiment, the stress is abiotic or biotic stress.

[0282] In one embodiment, the abiotic stress is selected from osmotic stress, oxidative stress, heavy metal stress, drought, submergence, cold, UV radiation, heat, ozone, nutrient deficiency and / or salt stress.

[0283] In another aspect of the invention there is provided a method for obtaining the genetically modified plant as described herein, the method comprising: a. selecting a part of the plant; b. transfecting at least one cell of the part of the plant of paragraph (a) with the nucleic acid construct as described above; c. regenerating at least one plant obtained from the transfected cell or cells; selecting one or more plants obtained according to paragraph (c) that show an altered stress response.

[0284] Further aspects and embodiments of the invention will be apparent to those skilled in the art given the present disclosure including the following experimental exemplification.

[0285] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. While the foregoing disclosure provides a general description of the subject matter encompassed within the scope of the present invention, including methods, as well as the best mode thereof, of making and using this invention, the following examples are provided to further enable those skilled in the art to practice this invention and to provide a complete written description thereof. However, those skilled in the art will appreciate that the specifics of these examples should not be read as limiting on the invention, the scope of which should be apprehended from the claims and equivalents thereof appended to this disclosure. Various further aspects and embodiments of the present invention will be apparent to those skilled in the art in view of the present disclosure.

[0286] All documents mentioned in this specification are incorporated herein by reference in their entirety, including any references to gene accession numbers and references to patent publications.

[0287] “and / or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. For example, “A and / or B” is to be taken as specific disclosure of each of (i) A, (ii) B and (iii) A and B, just as if each is set out individually herein. Unless context dictates otherwise, the descriptions and definitions of the features set out above are not limited to any particular aspect or embodiment of the invention and apply equally to all aspects and embodiments which are described.

[0288] The aspects of the invention are described in the following non-limiting examples.

[0289] Examples

[0290] Example 1 : Identification and ratification of DMSP synthesis genes from Spartina anglica

[0291] We first set out to identify candidate DMSP synthesis genes in Spartina anglica (Fig. 1 a), which is xenonative to the UK, an invasive coloniser of coastal wetlands worldwide9, and hosts key DMSP catabolic bacteria16. Spartina produces DMSP from the amino acid L-methionine via a methionine methylation pathway17(Fig. 1 b) that comprises four sequential enzymes: Methionine S- methyltransferase (termed MMT), S-methylmethionine (SMM) decarboxylase (termed SDC), DMSP- amine oxidase (termed DOX) and DMSP-aldehyde dehydrogenase (termed ALDH). Analysis of mRNA sequencing from triplicate S. anglica leaves, which contained 6 pmol g1FWDMSP, identified two transcripts from different loci that encoded proteins, SaMMTI and SaMMT2. These proteins shared >84% amino acid identity to the MMT enzyme from Zea mays. MMT is ubiquitous in plants18, catalyses the production of SMM as part of the SMM cycle19and is the first step of plant DMSP synthesis (Fig. 1 b). SaMMT2, which contained a 10 amino acid insertion at the conserved S- adenosylmethionine-binding domain (Fig. 2), showed no MMT activity and, thus, is not involved in SMM cycling or DMSP production. In contrast, SaMMT 1 , which contained no insertion in this domain, readily demonstrated in vitro MMT activity (Fig. 2). Using the S. anglica transcriptome, we also identified 11 transcripts encoding candidate aldehyde dehydrogenases highly homologous (75% protein identity) to characterised enzymes, notably betaine aldehyde dehydrogenase (BADH1 and BADH2) that were previously shown to have significant ALDH activity20. However, since aldehyde dehydrogenases are promiscuous in their substrate range, readily convert DMSP-aldehyde into DMSP in species that do not accumulate DMSP to high concentrations20, and (like MMT) are ubiquitous in plants, neither MMT nor ALDH are indicative of high-level production of DMSP in plants. This left the Spartina SDC and DOX enzymes as specific to plant DMSP synthesis17and as the best candidates for determining high-level DMSP production, especially as neither enzyme had been identified in a plant.

[0292] The S. anglica transcriptome was screened for genes encoding amino acid decarboxylases and copper amine oxidases, since SDC and DOX were previously predicted to belong to these enzyme families, respectively17. This identified three candidate SDC enzymes predicted to encode an arginine decarboxylase (SaADC), ornithine decarboxylase (SaODC) and diaminopimelate decarboxylase (SaDAPDC). SaDAPDC shared the highest identity to bacterial SMMDC and Burl proteins (with 27% and 24% amino acid identity, respectively) that catalyse this reaction in DMSP producing bacteria21 22. When expressed in and purified from E. coll, SaADC and SaDAPDC both showed in vitro activity with their predicted native substrates (arginine and diaminopimelate, respectively; Fig. 3a). However, when provided with SMM, only SaODC demonstrated SDC activity (Fig. 1 c; Fig. 3b, c), showing a Kmof 2.13 ± 0.20 mM and a Kcat of 1 .67 ± 0.08 nmol pg1min1protein (Fig. 3d). Contrastingly, SaODC demonstrated virtually no activity with ornithine (Fig. 3e, f). Additionally, SMM was roughly 14-times more abundant than ornithine in S. anglica leaf extracts. Therefore, we subsequently refer to the SaODC enzyme as SaSDC.

[0293] Two candidate DOX enzymes (copper amine oxidases) were identified in S. anglica that shared 30% amino acid identity and were termed SaCAOl and SaCAO2. Recombinant SaCAOl and SaCAO2 proteins were assayed for DOX activity, using pig kidney diamine oxidase as a positive control17. Both SaCAOl and SaCAO2 showed DOX activity, but the latter was ~45-fold more active (Fig. 1d), with an approximate Kcat of 298.57 pmol pg1min1(compared to 6.63 pmol pg1min1for SaCAOl). Furthermore, these enzymes were both independently identified by mass spectrometry analysis of the most active protein fractions from chromatographic experiments that enriched for S. anglica DOX activity (Fig. 4), consistent with them (particularly SaCAO2) being the enzymes responsible for DOX activity in this plant. As well as DMSP amine, SaCAO2 could use putrescine as a substrate and its high DOX activity was decreased by 97% in the presence of 20-fold excess putrescine, consistent with what is seen in S. anglica protein extracts (Fig. 1d), and previous work on DOX activity in Spartina extracts17. SaCAOl showed no activity towards putrescine. We therefore refer to SaCAO2 as SaDOX for the remainder of this study.

[0294] We next assessed the expression of SaMMTI, SaCAOl, SaDOX and SaSDC across different S. anglica tissues by RT-qPCR. Transcript levels of these genes, but most prominently, SaSDC, were significantly higher in leaves than any other tissue type (Fig. 1 e). Furthermore, SDC and DOX enzyme activities were also both highest in leaves (Fig. 1f and 1g), where plant DMSP synthesis was previously reported to occur23. Indeed, SaSDC transcripts and corresponding SDC enzyme activity were virtually undetectable or undetected, respectively, in any tissue besides leaf. Despite this, DMSP was still detected at high concentrations in other tissues, at 16-58% of the concentration in leaves (Fig. 1 h). Since SDC transcripts and activity were exclusively found in leaves (Fig. 1 e and f), our findings are consistent with DMSP being synthesised in the leaves and then subsequently mobilised throughout the plant to other tissues. It is also possible that some DMSP-amine synthesised in leaf tissue was exported to other tissues where it could be converted to DMSP, since DOX activity was only 21-42% decreased in non-leaf extracts (Fig. 1g). Whether this non-leaf DOX activity is due to SaCAOl , SaDOX and / or other unknown enzymes requires future investigation.

[0295] Example 2: DMSP accumulation in Spartina anglica is associated with abiotic stress

[0296] Spa / Y / na-populated saltmarshes are significant sources of climate-active gas emissions via DMSP catabolism10, yet little is currently known about how synthesis and accumulation of DMSP varies across natural Spartina populations, or what regulates DMSP levels in them. To address this, we measured DMSP concentrations in leaf samples from clumps of S. anglica growing across a transect in the saltmarsh at Stiffkey, Norfolk, UK (Fig. 5a; Fig. 6a). Our sampling strategy targeted a natural population across a single saltmarsh, which represents relatively limited genetic diversity (since S. anglica primarily undergoes vegetative propagation and the species formed from a hybridisation event some 200 years ago, resulting in a genetic bottleneck69), thus allowing us to focus on uncovering environmental (rather than genetic) factors that control DMSP accumulation. Unexpectedly, DMSP accumulation was highly variable, with differences as great as 14-fold (Fig. 5b). This is particularly striking given the magnitude of DMSP levels (>0.4-6 pmol g1FW). Moreover, there was no correlation in DMSP concentrations between clonal clumps growing within the same sampling site.

[0297] RNA-seq and differential expression analysis of the three lowest and three highest DMSP accumulating clumps (Fig. 6b) revealed the highest DMSP accumulating plants to have strongly elevated expression of an ethylene-responsive transcription factor 1 (EREBPT) homolog (Fig. 5c), a transcription factor whose expression has been linked to drought, submergence and pathogen stresses in other grasses2425. This upregulation suggested that these highest DMSP accumulating plants were experiencing elevated levels of one of these stresses, and concurrently had elevated concentrations of DMSP; DMSP synthesis may also be regulated by EREBP1 . Expression of MMT, SDC and DOX did not correlate with standing stocks of DMSP (Fig. 6c), suggesting that these genes may be post-translationally regulated, that DMSP is further metabolised26or that DMSP synthesis is influenced by substrate partitioning / availability27. Gene ontology enrichment analysis of differentially expressed genes (Fig. 5d) demonstrated a significant enrichment in high accumulators for biological process terms relating to oxidative stress and reactive oxygen species, salt and osmotic stress, and cellular detoxification, in agreement with the hypothesis that DMSP synthesis is stress regulated28. There was also a substantial enrichment of biological process terms relating to the metabolism of proline (Fig. 5d), a known osmolyte in plants29, suggesting a coupling with DMSP synthesis. To confirm this connection with proline metabolism, we measured proline, glutamate, and ornithine concentrations in S. anglica leaf tissues, in conjunction with a selection of other amino acids to contextualise DMSP accumulation within primary metabolism. We also assayed a variety of nutrients (N, S, K, P, Mg, Se, Mo, Fe, Zn) and ions and heavy metals known to contaminate saltmarshes (Na, Cd, Ni, Cu, Pb, Sr, Mn, Ba), both in leaf tissue (Fig. 5e) and soil (Fig. 6c), to determine if variation in these parameters influenced DMSP accumulation in the environment. Consistent with literature30, a negative correlation was observed between nitrogen levels and DMSP. We found that proline also negatively correlated with DMSP (correlation of proline with nitrogen was just short of the p-value cut-off; p = 0.08) (Fig. 5e). Leaf DMSP content also negatively correlated with soil nitrogen (Fig. 6d). Interestingly, DMSP positively correlated with the levels of molybdenum in leaves (Fig. 5e). This effect appears to be unrelated to the amount of molybdenum in soil, as this relationship is lost when correlated with total soil Mo (Fig. 6d). This is likely related to nitrogen deficiency, as Mo is used as a cofactor for nitrate reductase31, and nitrogen uptake is a demand driven process. It is also possible that S. anglica sulfate transporters contribute to Mo accumulation, as in other plants70. Consistent with this, S. anglica SULTR2;2 and SULTR 2;6 showed induced expression in the root (Fig. 16a, 16b), whilst the highest DMSP-accumulating plants had higher expression levels of SULTR3;4, SULTR3;6 and SULTR3;7 (Fig. 16c). There is a strong positive relationship between methionine and most other core amino acids (Fig. 5e), as can be expected given their connection via primary metabolism. DMSP does not share a relationship with these amino acids, indicating its decoupling from primary metabolism, which is also seen for SMM. We also observed no relationship between methionine, SMM and DMSP levels (Fig. 5e), despite the former two being synthesis intermediates. This suggests that the regulation of flux occurs at a point after MMT, and that these amino acid pools are resilient to depletion by large fluxes of DMSP synthesis in S. anglica.

[0298] To test how DMSP accumulation is regulated by salt or sulfur status, we performed experiments with glasshouse-grown S. anglica plants watered with freshwater, salt solution (NaCI), or sea salts (containing NaCI and high concentrations of sulfate). Interestingly, DMSP accumulation was not significantly increased in plants watered with NaCI relative to plants treated with freshwater (consistent with 30,71), but plants watered with sea salts did contain higher DMSP concentrations (Fig. 16d). This finding suggests that DMSP accumulation is not regulated by NaCI treatment, but rather is regulated by sulfate availability. Consistent with this, in the natural S. anglica populations we found no correlation between DMSP accumulation and Na concentrations in the leaf (Fig. 5e) or soil (Fig. 6d). Furthermore, measurements of sulfate and O-acetylserine (a key sulfur status indicator72) in leaf material from the three lowest and three highest DMSP accumulating clumps revealed no significant differences in the concentrations of these compounds (Fig. 6e), suggesting that DMSP accumulation places no significant drain on the sulfur reserves of S. anglica. Overall, these findings support a role for DMSP synthesis being an adaptation of S. anglica to growing in a sulfur-rich saltmarsh environment. Taken together, these data demonstrate that DMSP accumulates dynamically across populations of S. anglica in the environment. The level of accumulation depends on nitrogen availability, and the contrasted transcriptional profile of highest and lowest DMSP accumulators suggests a link with abiotic stress, supporting the role of DMSP as an anti-stress molecule. DMSP accumulation is increased in response to elevated concentrations of sulfate but not salt. Additionally, the lack of correlation between transcript abundance of synthesis genes and standing stocks of DMSP suggest more complex regulation than simple up or downregulation of gene expression.

[0299] Example 3: DMSP synthesis is widespread in plants and evolution of high-level production is convergent

[0300] To determine the breadth of DMSP production in plants, DMSP concentrations were measured in phylogenetically and environmentally diverse species spanning the plant kingdom. Consistent with literature14 15, all 43 tested plant species produced DMSP, albeit mostly at levels that were two or four orders of magnitude lower than in Saccharum officinarum or Spartina anglica, respectively (Fig. 7a). No new high-level DMSP accumulating plants were identified and this trait should therefore be considered rare. However, it should be noted that species were identified which accumulated DMSP concentrations only 5-times lower than Saccharum officinarum; these were Araucaria araucana (monkey puzzle; 125 nmol g1FW) and Eurhynchium striatum (common striated feather-moss; 113 nmol g1FW). DMSP accumulation at lower levels was universal in plants, implying that terrestrial systems may be a significant source of DMSP. There was no clear phylogenetic link between species that demonstrated high-level DMSP accumulation, indicating that this characteristic likely evolved convergently multiple times. Furthermore, DMSP concentrations were previously shown to be elevated in Arundo donax and Solanum lycopersicum in response to drought14 15, suggesting that DMSP accumulation in other lower DMSP-producing plants may also be enhanced by conditions linked to climate change and potentially become more significant sources of DMSP and related climate-active gases under such future conditions.

[0301] To determine how plants accumulate different concentrations of DMSP, we assessed the prevalence and activity of Spartina DMSP synthesis enzyme homologs from different species. SaMMTI in vitro activity was comparable to previously characterised plant MMTs from Arabidopsis thaliana and Hordeum vulgare, which accumulate low concentrations of DMSP (Fig. 2a). Phylogenetic analysis also showed plant MMT diversity to follow taxonomy, with MMT from high accumulating species not being distinct from those from low producers (Fig. 8a).

[0302] High-level SDC activity was previously shown to be rare in plants17. Proteins homologous to SaSDC (50-84% amino acid identity) were also identified in most higher plants (Fig. 7b), but notably not in A. thaliana, which lacks ornithine decarboxylases32, nor from the available RNA-seq data from the high DMSP producer P. oceanica33nor in the genome sequence of Zostera marina, implying the existence of (an)other DMSP synthesis route(s) in these species. The proteins homologous to SaSDC from the high-level DMSP producer Saccharum officinarum and low-level DMSP producers Solanum lycopersicum, Setaria viridis and Spartina patens, all converted ornithine into putrescine (Fig. 8b), but significantly, lacked SDC activity (Fig. 7c). Despite these stark differences in enzyme activity, SaSDC phylogenetically clustered with the other ODC enzymes in a manner reflective of phylogeny (Fig. 7b). Sequence analysis did not highlight any substantial amino acid insertions or deletions between SaSDC and those ODC enzymes lacking SDC activity to explain the gained activity of SaSDC on SMM substrate, implying this difference is likely due to a small number of specific amino acid substitutions. Together, these data imply that biochemical changes in SaODC that alter its function to that of an SDC, underlie evolution of high-level DMSP accumulation in S. anglica. We also conclude that Saccharum officinarum and seagrasses P. oceanica and Z. marina, must use distinct enzymes / pathways to synthesise and accumulate DMSP at high concentrations.

[0303] Proteins with 87-91% protein identity to SaDOX were ubiquitous in the plant kingdom (Fig. 7d) and, like MMT, showed much less diversity than seen in the SDC / ODC family enzymes (Fig. 7b; Fig. 8a). Indeed, SaDOX homologs from S. officinarum and Setaria viridis had DOX activity equivalent to the S. anglica DOX, and Solanum lycopersicum had lower activity (30%) (Fig. 7e), implying that enzymes with DOX activity were themselves not determinants of high-level DMSP accumulation. This seemingly contradicts previous work demonstrating enhanced DOX rates in Spartina species that accumulated high DMSP concentrations compared to plants now known to accumulate low DMSP concentrations17. We subsequently assayed protein extracts from S. anglica, S. patens, Solanum lycopersicum, Nicotiana benthamiana, A. thaliana and H. vulgare for DOX activity, species for which we were able to identify DOX homologs (with the exception of Spartina patens, for which no genetic resources exist and forwhich we were unable to clone a DOXbased on sequences designed against S. anglica DOX). As reported17, S. anglica showed much higher levels of DOX activity (12-times higher) than the low producing species (Fig. 8c), and this activity was also inhibited by excess putrescine. To compare expression levels of the DMSP synthesis genes in these species, we analysed publicly available transcriptomic datasets (A. thaliana, H. vulgare and N. benthamiana) or datasets generated in this study (S. anglica and Solanum lycopersicum). In these data, we identified homologs of MMT, SDC, DOX, CAO1 (as well as ACTS and EF1A to correct for sequencing depth artefacts) and calculated the transcript abundance for each relative to ACT8 (Fig. 7f). This revealed that DOX is much more highly expressed in Spartina anglica relative to the other low producing species (33-84-times). MMT transcript levels were equivalent between S. anglica and the other species, except, unexpectedly, in H. vulgare, which demonstrated highly elevated levels of MMT. SDC levels in S. anglica were roughly double those of other species. Overall, these data demonstrate that DOX activity is common in plants, and there is little enzymatic difference between DOX from high producing S. anglica and other low producing species. However, in S. anglica, DOX is much more highly expressed, and therefore DOX activity is substantially higher overall. Example 4: DMSP functions in plant stress and can improve stress tolerance

[0304] To determine how DMSP functions in plants, we used tomato as a model, since this species produces DMSP in a salinity- and drought-regulated manner14and is highly amenable to laboratory experiments. We subjected plants to salt stress in the presence / absence of DMSP. DMSP was taken up by the roots and transported into aerial tissues, resulting in DMSP accumulation in the leaves 4- 6-fold higherthan in untreated plants (up to maximum levels of 21 nmol g1FW; Fig. 9a). As expected, salt stress resulted in significantly decreased biomass (42% lowerthan control plants; Fig. 10a, 10b). Strikingly, addition of DMSP resulted in total biomass rescue (Fig. 10a, 10b), demonstrating for the first time the protective effects of this compound in plants. We also subjected tomato plants to osmotic stress (using the osmoticum sorbitol) in the presence / absence of DMSP, and observed a 50% increase in total biomass in plants supplemented with DMSP relative to those plants exposed to the osmotic stress alone (Fig. 11). This result demonstrates that DMSP is not only protective against salt stress, but also osmotic stress.

[0305] To dissect further the mechanisms by which DMSP acts in planta, we performed RNA-seq on plants grown in the presence of salt and / or DMSP (Fig. 9b). Salt stress resulted in large-scale transcriptomic changes relative to control plants (Fig. 9c), with 1033 transcripts differentially expressed by 2-fold or greater in NaCI treated plants compared with control plants. In NaCI and DMSP treated plants, this number was decreased to 704 transcripts, of which 427 were also differentially expressed between NaCI treated plants and the untreated control. Gene ontology enrichment on genes that were differentially expressed in response to DMSP treatment alone identified enrichment in processes relating to water movement and auxin signalling (Fig. 9d), suggesting association with salt stress responses34. A total of 175 genes were differentially expressed in plants treated with NaCI and DMSP (Fig. 10c) relative to NaCI treated plants. Gene ontology enrichment revealed that many of these genes were involved in developmental processes, as expected given NaCI treated plants were developmentally arrested whilst NaCI and DMSP treated plants were not (Fig. 10a, b). Of these genes, many of the most differentially expressed (or their homologs in other species) have roles in osmotic stress (SISWEET, SIACO, SIFIBRILLIN, SITIP3;2)35-38, oxidative stress (SIGLO2, SIFER2, SIOPT3, SILEA5, SIERF105)39 43or salt / metal stress (SITDT, SIPME13, SIPCR2, SIREC2, SISTZ)44-48(Fig. 10d). These genes may therefore represent pathways through which DMSP exerts its protective effect. Taken together, these data demonstrate that DMSP can ameliorate salt stress, and likely does so in tomato by counteracting osmotic stress and oxidative stress. This aligns well with our S. anglica RNA-seq (Fig. 5c), suggesting DMSP likely acts in a conserved way across species.

[0306] To determine if the application procedure altered the effectiveness of DMSP on plant stress tolerance, we tested two distinct application methods. The first involved applying a single application of DMSP through watering / root drenching of tomato plants growing in soil, and then measuring DMSP content in the leaves of these plants, plus DMSP content in the surrounding soil. In this experiment, soil watered with solutions containing 50 or 500 pM DMSP showed relatively limited retention of DMSP (typically up to c. 2 weeks post application; Fig. 12a), with higher concentrations detected in soil watered with the solution containing more DMSP. In contrast, DMSP concentrations in the leaves were highly elevated (reaching 50-150 nmol g1FW for the 500 pM DMSP treatment), and to our surprise, these elevated concentrations persisted for many weeks post the original application of DMSP (Fig. 12b, Fig. 19a). Importantly, no obvious visible detrimental effects were observed on plant growth in response to the DMSP treatment (Fig. 12c). This significant finding demonstrates two important things: i) that DMSP is taken up by the plant roots, where it can be subsequently transported to the leaves, and ii) that this systemic accumulation of DMSP in the leaves persists long after the initial application. We saw the same effect when barley plants were watered with a solution containing DMSP, with DMSP concentrations in the leaves reaching 20-180 nmol g1FW for the 500 pM DMSP treatment and accumulation lasting for many weeks after the original application of DMSP (Fig. 19b). We then tested whether plants watered with DMSP showed improved stress tolerance, and indeed, tomato plants exposed to salt stress and watered with DMSP were larger than plants exposed to salt stress alone (Fig. 13a). Furthermore, the leaves of these salt- stressed tomato plants watered with DMSP were healthier than the leaves of plants treated with salt stress alone, as determined by visual appearance and quantifying total chlorophyll content (Fig. 13b, c). Interestingly, plants watered with DMSP alone (in the absence of salt stress) showed some early flowering and increased internode length (Fig. 13a). Our second method test involved a single foliar application of DMSP as a topical spray to the leaves of tomato plants growing in soil. In this experiment, the leaves were sprayed with a solution containing 500 pM DMSP and then, after three days, exposed to oxidative stress (through application of the herbicide Paraquat). The control plants (i.e. non-DMSP treated) showed yellowing / browning of the leaves, but surprisingly the plants treated with DMSP were greener, as determined by quantifying total chlorophyll content (Fig. 14). Together, these important findings demonstrate that DMSP can be applied to plants growing in the soil via two distinct methods, and that DMSP can have protective effects against different stresses for plants growing in soil.

[0307] To explore further the effectiveness of spray application of DMSP, barley plants were sprayed with a single application of solutions containing different DMSP concentrations. DMSP content in nonsprayed leaves was measured after the initial application to see whether DMSP would be taken up by the plants and then remain in the plant tissue. To our surprise, elevated concentrations of DMSP were detected in non-sprayed leaves for many weeks / months after the initial application (Fig. 20), demonstrating that DMSP applied by spraying is indeed taken up by plants, transported to other plant tissues (including newly growing aerial tissue) and accumulates in these tissues for significant periods oftime after being applied. Application of solutions containing higher concentrations of DMSP resulted in higher final concentrations detected in the leaves and showed greater persistence over time (Fig. 20a, b), with 50 mM applications resulting in elevated leaf DMSP concentrations for at least 9 weeks (Fig. 20a). Excitingly, these spray treatments also resulted in barley plants with increased biomass, with the 50 mM DMSP application resulting in a 59% increase in total biomass relative to the control unsprayed plants (Fig. 21 a). Importantly, when these barley plants were also subjected to salt stress (watering with 100 mM NaCI), the plants treated with DMSP showed increased biomass relative to control plants not sprayed with DMSP (Fig. 21 b). The barley plants sprayed with 50 mM DMSP showed a 38% increase in total biomass relative to unsprayed plants (Fig. 21 b). Spray applications of DMSP at lower concentrations (1 and 5 mM) showed increased biomass relative to unsprayed plants under both normal conditions and salt stress conditions, but larger increases in biomass were seen for the spray treatments with higher DMSP concentrations (e.g. 50 mM DMSP; Fig. 21).

[0308] We then tested whether the spray application of DMSP could be translated to another crop and another stress, focussing on pea and drought stress. There was no observable difference between untreated pea plants and those treated with a single 50 mM DMSP spray application under well- watered conditions, but there were significant improvements for the sprayed plants under drought stress (Fig. 22). Pea plants sprayed with 50 mM DMSP showed a 176% increase in biomass relative to unsprayed plants (Fig. 22a) and had almost twice as much chlorophyll content (Fig. 22b). Remarkably, DMSP content in the leaves of the sprayed pea plants reached 800-1200 nmol g1FW (Fig. 22c), levels which were comparable with the saltmarsh cordgrass Spartina anglica (Fig. 5b). The improved growth of drought-stressed pea plants sprayed with DMSP was clearly visible when compared to unsprayed plants (Fig. 22d).

[0309] We next tested whether these findings could be translated from controlled environment facilities to the glasshouse, using wheat as a test species. Application of DMSP by watering onto the soil (root drench) or spraying directly onto wheat plants both resulted in elevated concentrations of DMSP in leaf tissue that had not come into direct contact with DMSP upon the initial treatment (Fig. 23a). As with barley, higher DMSP application concentrations resulted in higher DMSP concentrations detected in the leaves of the plants, and accumulation was seen for many weeks / months after the application was made. In these glasshouse experiments, three DMSP applications were made to the plants (3 weeks apart), and this resulted in significantly higher maximum concentrations of DMSP detected in the plants, e.g. a single 50 mM DMSP spray in barley resulted in DMSP leaf concentrations of 5-10 nmol g1FW being achieved 7 weeks after application (Fig. 20), whilst three 50 mM DMSP sprays in wheat resulted in DMSP leaf concentrations of 100-500 nmol g1FW being achieved 7 weeks after application (Fig. 23). This suggests that multiple applications of DMSP (by either spraying or root drench) result in greater accumulation of DMSP in plant tissues overtime (Fig. 23). The leaf concentrations of DMSP achieved in wheat with the higher application concentrations (50 and 100 mM DMSP) in the initial weeks following spraying (Fig. 23a) again resulted in levels comparable with S. anglica (Fig. 5b), mirroring what was observed in pea (Fig. 22c). The wheat plants sprayed with 100 mM DMSP also had 11 % more chlorophyll than control (unsprayed) plants (Fig. 23b), demonstrating that the increases in chlorophyll content resulting from DMSP application can be translated from plants grown under controlled environments (Fig. 13c, Fig. 14) to plants grown under glasshouse conditions. When the wheat plants sprayed with DMSP were also subjected to drought stress, we observed significant increases in total biomass (38% more fresh weight in plants sprayed with 100 mM DMSP vs. control unsprayed plants; Fig. 24a) and chlorophyll content (22% more chlorophyll in plants sprayed with 100 mM DMSP vs. control unsprayed plants; Fig. 24b), demonstrating that DMSP application improves drought stress tolerance in wheat as well as other plant species. To test if these results could be further translated to the field, field-grown wheat plants were sprayed either two or three times with DMSP solutions of different concentrations. Nine weeks after the final application, DMSP concentrations in the leaves were measured and all three treatment regimens resulted in increased DMSP leaf content relative to control unsprayed wheat plants (Fig. 25), demonstrating that DMSP applications can be made successfully to plants grown in the field.

[0310] Finally, we set out to demonstrate that DMSP synthesis is a trait which can be engineered in plants. Transient expression of SaMMTI, SaSDC and SaDOX alone and in combination in N. benthamiana demonstrated that over-expression of all three genes was required to elevate DMSP concentrations significantly (Fig. 10e). Importantly, these transiently transformed plants with elevated DMSP accumulation (23 nmol g1) demonstrated drought resistance relative to control plants (Fig. 9e). Transgenic A thaliana plants over-expressing SaMMTI, SaSDC and SaDOX were also generated, and the resulting lines accumulated DMSP up to 3.4 pmol g1FW (Fig. 10f), i.e. concentrations of DMSP equivalent to S. anglica (Fig. 5b). We also generated transgenic barley plants expressing SaMMTI, SaSDC and SaDOX, with several of the resulting lines accumulating DMSP between 1 and 34 pmol g1FW (i.e. concentrations 10-100 fold greater than those detected in S. anglica; Fig. 15a). A., thaliana seedlings of the second highest DMSP synthesising line (1.3 pmol g1FW) and wildtype Col-0 (10 nmol g1FW) were grown and subjected to NaCI stress. These transgenic plants did not demonstrate any growth defects, and, excitingly, when exposed to 100 mM NaCI, demonstrated a significant increase in biomass relative to control plants (Fig. 10g and h; Fig. 15b), similar to what was seen through root uptake of DMSP (Fig. 10a; Fig. 11 ; Fig. 12). Moreover, the transgenic barley lines showed improved tolerance to drought stress (Fig. 26), corroborating the result seen for transient expression of SaMMTI, SaSDC and SaDOX in N. benthamiana (Fig. 10e) and the work with topical application of DMSP (e.g. Fig. 22, Fig. 24). Overall, these data demonstrate that DMSP levels can be manipulated in low DMSP-accumulating species through root uptake or spraying of DMSP or by over-expression of MMT, SDC, and DOX from S. anglica, and that doing so can increase tolerance to salt stress and drought.

[0311] Discussion

[0312] In saltmarshes, cycling of Spartina derived DMSP is considered to be the key source of climate active DMS10. All plants tested both here and in other studies produce DMSP (Fig. 7a;14 15), though many do so at low concentrations. Many of these low DMSP-accumulating species are crop plants that cover large areas; for example, in 2022 in the UK, wheat and barley covered 1 ,809,000 and 1 ,104,000 hectares, respectively49. Cumulatively, terrestrial environments may therefore play host to significant DMSP cycling, a characteristic previously thought to be confined exclusively to marine environments. Indeed, one would predict that environments associated with plants that accumulate high DMSP concentrations contain higher proportions of microorganisms able to catabolise DMSP, and thus produce more DMS, than those from plants accumulating low DMSP concentrations. To test this hypothesis, we analysed a recently published metagenomic dataset of sediments associated with S. alterniflora (high DMSP producer) and S. patens (low DMSP producer;73). Although there were some differences in the abundance of specific DMSP lyase genes, notably in dddY, surprisingly, there was no significant difference between the high and low DMSP accumulators in the cumulative abundance of known DMSP lyase genes (15.9 ± 2.4 vs 20.5 ± 3.7%) or the DMSP demethylation gene (8.3 ± 1.7 vs 8.6 ± 1.3%) (Fig. 17). The reason for this could be because: the transcript and protein abundance for these genes and the DMSP catabolic rates (not examined in73), differed between these samples; S. alterniflora retains DMSP for its important physiological roles; or other organisms (e.g. bacteria) provide the DMSP to this environment (as implied by74). Importantly, these terrestrial DMSP lyase gene abundances are very similar to those previously reported for the Earth’s surface oceans75and coastal surface marine sediments74. The significance of terrestrial DMSP cycling thus demands further investigation to determine the significance of these processes in diverse environments, particularly in the context of other microbial activities, e.g. carbon fixation and nitrogen fixation as recently identified in the microbiome of S. alterniflora76.

[0313] Our identification and characterisation of the genes presented here constitute the first molecular genetic study of DMSP synthesis in plants. MMT and ALDH (Fig. 1 a) are ubiquitous in plants1720regardless of how much DMSP they produce, and as such are not clear determinants of high-level DMSP production. MMT enzyme activity and gene expression is equivalent between low and high DMSP producing plants (Fig. 2a; Fig. 7f). DOX genes also appeared near-ubiquitous in plants, showed little variation (Fig. 7d), and were enzymatically similar between species (Fig. 7e). However, in S. anglica, this gene is subject to elevated expression (Fig. 7f), resulting in overall much greater DOX activity (Fig. 8c). Importantly, our work demonstrates that SDC is the most critical enzyme for evolution of high-level DMSP synthesis in Spartina, with the S. anglica SDC demonstrating markedly different kinetics with SMM than homologous ODC enzymes from other plant species (Fig. d). Indeed, this difference was salient when comparing to the ODC of S. patens, a species that diverged fewer than 2-4 MYA from S. anglica50but that fails to produce high concentrations of DMSP (Fig. 7a). In addition to highlighting the importance of evolution of ODC to SDC for high-level DMSP-synthesis in the Spartina lineage, this comparison also provides a timeframe in which this change occurred. Additionally, the lack of detectable SDC activity or expression in other high DMSP producers (Saccharum officinarum, P. oceanica, Z. marina) suggests the existence of other plant DMSP synthesis routes that are SDC-independent (e.g. as in M. biflora5V), which warrant future investigation. Overall, we conclude that high-level DMSP synthesis evolved convergently multiple times in plants. DMSP is an anti-stress molecule and levels of DMSP in S. anglica growing in the environment showed substantial variation, which was unlinked to the primary metabolic state of plants (Fig. 5). DMSP concentrations showed a positive correlation with plant molybdenum stores, and a negative correlation with nitrogen and levels of proline (Fig. 5e), a protein-coding amino acid that also serves as an osmolyte and anti-stress molecule in plants52. Given the highly saline nature of saltmarshes, S. anglica must maintain elevated pools of osmolytes, but this represents a substantial drain on nitrogen reserves, which are frequently limiting in Spartina species53. Sulfur is rarely limiting in saltmarshes owing to daily fluxes of sulfates from tides54, so DMSP likely represents a component of a diverse toolkit of osmolytes that S. anglica can draw upon to maintain its water potential when met with specific nutrient limitations. Additionally, as ammonia is released from DMSP-amine to produce DMSP-aldehyde (Fig. 1 b), this synthesis pathway could ultimately also be used to recycle nitrogen from methionine pools, thereby helping to balance the nitrogen and sulfur demands of the plant.

[0314] By modulating DMSP levels through root uptake or over-expression of MMT, SDC and DOX, we demonstrate that DMSP can improve the tolerance of plants to salt stress and drought (Fig. 10). The mechanism of action of DMSP was likely through its capacity to act as an osmolyte and counteract oxidative stress (Fig. 10d). Such an approach may be of particular benefit in nitrogen-poor soils, where production of nitrogen-containing osmolytes would be a significant drain on the limited nitrogen levels of a plant. Manipulation of DMSP levels in plants through root uptake and the use of transgenics to engineer DMSP production represent potential routes to enhance abiotic stress resistance and improve agricultural productivity. Whether transgenic plants with elevated levels of DMSP would also significantly alter DMS flux into the atmosphere remains to be explored, but this would be an exciting area of future study, particularly in the face of global climate change.

[0315] Methods

[0316] Spartina anglica sampling

[0317] S. anglica leaf tissue was collected from Stiffkey saltmarsh in Norfolk, UK (Fig. 6A). Tissue was washed three times using deionised water to remove surface contaminating bacteria and then RNA and protein was extracted immediately for transcriptomics and protein activity assays. For each plant, soil was also collected from the base of the plant for ICP-AES and CHN analyses.

[0318] RNA extraction from Spartina anglica and assembly of transcriptome

[0319] For each S. anglica sample, leaves were collected from 4-6 clumped individuals. These were then cut into segments, pooled, and 1 g was homogenised in liquid nitrogen. RNA was extracted using Tri Reagent Solution (ThermoFisher Scientific, AM9738) as per the manufacturer’s instructions. RNA was cleaned up using Zymo RNA Clean & Concentrator™-25 (Zymo-Research). Extracted RNA was then sent to Novogene Co. where mRNA libraries were prepared. The mRNA libraries were sequenced with paired-ends at a read depth of approximately 20 million reads. Adapters were trimmed from reads using Trim Galore55and a reference transcriptome was built using Trinity56under default settings. For highest and lowest DMSP accumulators, a reference transcriptome was constructed using both sets of samples, and then redundancy was removed.

[0320] Identification of candidate genes from Spartina anglica transcriptome

[0321] For each gene of interest (MMT, SDC, DOX and ALDH), a list of protein sequences containing the domain of interest was curated for A. thaliana, Z. mays and T. aestivum, using the InterPro entries IPR025779, IPR008286, IPR000269 and IPR015590, respectively. A tBLASTn search was run against the S. anglica transcriptome using each of the lists as query sequences. The contigs identified by this BLAST search were then subset so that only those with an e-value lower than 0.0005 were carried forward. For MMT, SDC and DOX lists, the non-redundant contig sequences were then manually translated and continuous reading frames with start and stop codons were extracted. For each open reading frame (ORF), a BLASTp search was run against the complete BLAST database, and only those which had near complete alignment (>90%) to matching genes of interest were carried forward. The ORFs were aligned against one another and redundant ORFs were discarded. Finally, the nucleotide sequences for each of the remaining ORFs were identified from the original contigs, and these were domesticated for Golden Gate cloning and synthesised via commercial DNA synthesis. This resulted in two transcripts for MMT, three for SDC and two for DOX.

[0322] Protein extractions from Spartina anglica

[0323] For DOX activity tracking, 60 g of S. anglica leaf tissue was used for protein extraction. The plant material was blended in 150 ml buffer containing potassium phosphate buffer 50 mM pH 8, DTT 5 mM, EDTA 1 mM, L-ascorbic acid 5 mM (Buffer A), for purification of DOX activity, as described previously17. Blended samples were filtered through three layers of Miracloth (Millipore), followed by dialysis with SpectraPor™ dry standard grade dialysis tubing MWCO 6-8000 (Repligen) overnight. Dialysis was performed in 5 litres of Tris 20 mM pH 8, NaC1 150 mM, which was changed three times. Samples were then centrifuged for 45 minutes at 4000g, and the supernatant was taken forward. Protein was then quantified by Bradford assay and brought to a final concentration of 2 mg ml1. All steps were performed at 4°C.

[0324] For all other enzyme extract experiments, 5-10 g of leaf tissue was used, and homogenisation was performed under liquid nitrogen using a mortar and pestle. For SDC assays, powder was thawed in potassium phosphate buffer 50 mM pH 7.2, DTT 5 mM, EDTA 1 mM, pyridoxal 5’-phosphate 0.1 mM, L-ascorbic acid 5 mM (Buffer B), centrifuged at 4000g for 10 minutes to clarify, and then filtered through Miracloth. Material for DOX assays was treated the same, but thawed in Buffer A. Filtered extracts were then desalted by PD-10 columns (Cytiva) using gravity filtration and exchanged into fresh Buffer B for SDC, and Buffer A with no L-ascorbic acid for DOX.

[0325] Isolation of DOX by activity tracking Protein extract was brought to 40% ammonium sulfate over 1 hour and was incubated at 4°C for 1 hour. The protein extract was centrifuged for 30 minutes at 4000g, and the supernatant was dialysed overnight using SpectraPor™ dry standard grade dialysis tubing MWCO 6-8000 (Repligen). Proteins were then fractionated on a phenyl sepharose column (Cytiva) with a programme of 0-100% PS Buffer A (Tris-HCI 50 mM pH 8, 20% Ammonium sulfate) against PS Buffer B (Tris-HCI 50 mM pH 8) over 400 ml. Two fractions demonstrated activity and the fraction with the highest activity was taken forward for Q anion exchange chromatography, in which the proteins were fractionated across a gradient of Q Buffer A (Tris-HCI 50 mM pH 8, NaCI 1 M) 0-100% against Q Buffer B (Tris-HCI 50 mM pH 8, NaCI 50 mM) over 400 ml. A single fraction demonstrated DOX activity. This final fraction was separated by gel filtration with Superdex HiLoad 16 / 600 Column (Cytiva) with an isocratic flow of Q Buffer B. After each fractionation, DOX activity demonstrated an enrichment (Fig. 4b). Two samples were sent for LC-MS analysis (Fig. 4b) at the John Innes Centre Proteomic Platform. These samples were the final active fraction after gel filtration chromatography and the lower activity fraction taken from the phenyl sepharose chromatography.

[0326] Purification of recombinant proteins from E. coli

[0327] Genes of interest were cloned into a pMALc2x plasmid modified for Golden Gate cloning and then transformed into E. coli strain BL21 DE3. Positive colonies (verified by Sanger sequencing) were grown overnight at 37°C 180 rpm in 10 ml Luria-Bertani (LB) broth containing 100 mg ml1carbenicillin. These 10 ml cultures were then used to inoculate 1 litre cultures (LB broth containing 100 mg mH carbenicillin and 0.05% glucose), which were grown at 37°C 180 rpm to an ODeoo of 0.6 and then induced with 0.5 mM IPTG at 28°C 180rpm for 3 hours. Cultures were then pelleted, resuspended in lysis buffer (Tris-HCI 20 mM pH7.4, NaCI 200 mM, EDTA 1 mM, DTT 1 mM) and lysed by French press at 10000 psi. Samples were separated into soluble and insoluble fractions by centrifugation at 9000g for 30 minutes, and the soluble fraction was then incubated for 1 hour with amylose resin (New England Biolabs E8021 S) pre-washed with washing buffer (Tris-HCI 20 mM pH 7.4, NaCI 200 mM, EDTA 1 mM) four times. We confirmed that each protein of interest was in the soluble fraction by running samples of total lysed extract, soluble fraction, and insoluble fraction on polyacrylamide gels. After incubation with amylose resin, samples were centrifuged for 1000g for 1 minute and the supernatant removed. The resin was washed four times with washing buffer, and then poured into an empty PD-10 column with filter (Cytiva). Wash buffer was allowed to run through by gravity, followed by a wash with washing buffer 2 (Tris-HCI 20 mM pH 7.4, NaCI 200 mM). Columns were then incubated for 5 minutes with 10 mM maltose (Merck 63418) dissolved in washing buffer 2, and the eluate was collected and quantified by Bradford assay.

[0328] Enzyme activity assays For total plant extracts 20-40 pg of protein was used, and for purified enzymes 2-10 pg of protein. All enzyme reactions were performed at 25°C. To assay MMT activity, protein extracts or purified enzymes were incubated with 1 mM L-methionine (Merck 1 .05707) and 1 mM S-adenosylmethionine (Merck 798231) in a reaction buffer containing Tris-HCI 20 mM pH 7.2 and DTT 1 mM.

[0329] Decarboxylase assays were incubated with 1 mM substrate (L-S-methylmethionine (Biosynth FM25160), L-ornithine (Merck 02375), L-diaminopimelic acid (Merck 33240) or L-arginine (Merck A5006)) in a buffer containing Tris-HCI 20 mM pH 7.2 and pyridoxal 5’-phosphate 0.1 mM (Merck P9255). For plant extracts, 5 mM L-ascorbic acid was also included in this mixture to inhibit removal of the product by DOX activity. Decarboxylases were measured by HPLC and NMR for the kinetics. Note; for experiments with SaSDC, 10 pg of protein was used for SDC assays, but 35 pg was used for ornithine assays, owing to a lack of detectable activity.

[0330] DOX assays were incubated with 1 mM substrate (unless otherwise specified), either putrescine (Merck 51799) or DMSP-amine which was synthesised enzymatically using our identified SDC and confirmed by HPLC. Reactions were set up in Tris-HCI 20 mM pH 8 in crimped 2 ml gas-tight glass vials and incubated at 25°C in the dark. As DMSP-aldehyde is unstable, it decays with a half-life of ~1 hour17, resulting in DMS in the headspace of these vials. Measurements by gas chromatography were then taken of headspace DMS from decayed DMSP-aldehyde after a minimum of 2 hours.

[0331] High Performance Liquid Chromatography (HPLC)

[0332] Enzyme assays were precipitated by the addition of 100% trichloroacetic acid (Merck T0699) to a final concentration of 0.15%. Samples were incubated at 4°C for 10 minutes and then centrifuged at 13000g for 10 minutes. The supernatant was transferred to a new tube and 50 pl of this supernatant was then reacted with an equal volume of ortho-phthaldialdehyde (Merck P0532) for 5 minutes for the derivatisation reaction to occur. The fluorescent adducts were then resolved via reverse-phase chromatography by injecting 20 pl of sample on to a Synergi™ 4 pm Hydro-RP 80 A 100x2 mm column (Phenomenex) using buffers A (NaH2PC>4 25 mM pH 2.5 adjusted with H2PO4) and B (50:40:10 methanol:acetonitrile:water), as described previously57, with a flow rate of 0.3 ml min1. For our method, we ran a gradient of 90% A and 10% B to 20% A and 80% B over 12 minutes, which was then held for 5 minutes. The column was then brought back to 90% A and 10% B over 2 minutes and allowed to equilibrate for a further 5 minutes. A Dionex RF2000 fluorescence detector (Dionex) was used to detect the products with excitation at 332 nm and emission at 445 nm.

[0333] For sulfate measurements, 10 mg of dry ground sample was extracted in 200 pl water for 30 min with sonication in a bath sonicator. The extract was diluted 100-fold in water, and 10 pl was injected onto a Dionex Ion Pac AS18 column 2 x 250 mm, fitted with AG18 2 x 50 mm guard column. The column was eluted with a gradient of potassium hydroxide delivered at a flow rate of 0.25 mL min-1 by a Dionex ICS-2100 Ion Chromatography System and Dionex AS-AP autosampler. The gradient was delivered with the schedule: time (min), mM; 0,12; 12,12; 20,34; 25,100; 25.1 ,12; 35,12. Sulfate was detected as a peak with retention time of 12.18±0.12 min. Calibration curves for injection of 10 pmol- 10 nmol gave R2>0.998 and peaks were integrated in Dionex Chromeleon v.6 software.

[0334] For O-acetylserine measurements, 10 mg of dry ground sample was extracted in 200 pl water for 30 min with sonication in a bath sonicator. The extract derivatised with ortho-phthaldialdehyde (Merck P0532), and samples were resolved as described for enzyme assay measurements with the following differences: 10 pl of sample was injected onto the column, and the gradient was delivered with the schedule: time (min), % B: 0,20; 20,80; 25,80; 26,20; 31 ,20. The retention time of O-acetylserine was 8.11 ± 0.05 min. Calibration curve for injection of 25-500 pmol gave R2=0.998, and peaks were integrated in Jasco ChromNav v.1 software.

[0335] Gas Chromatography (GC)

[0336] DMSP measurements were conducted using GC. For plant samples, a known quantity of tissue (usually between 30-50 mg) was placed in a 2 ml glass vial containing 0.1 ml deionised water. The sample was homogenised using a pipette tip, 0.1 ml of 10 M NaOH was added to each homogenate to liberate DMS from DMSP by alkaline lysis and followed by immediate crimping. Samples were incubated in the dark at room temperature overnight and assayed the subsequent day. DMS in the headspace of these vials was resolved using an HP-INNOWax 30 m x 0.530 mm capillary column and measured using a flame photometric detector (Agilent 7890A GC system fitted with a 7693 autosampler). For all species, the presence of DMSP was confirmed by LC-MS as described previously58.

[0337] Nuclear Magnetic Resonance (NMR)

[0338] All NMR measurements of enzyme reactions were performed in a 5 mm NMR tube at 298K on a Bruker500 MHz spectrometer with autosampler. Product formation was quantified against an internal standard of 1 mM pyrazine (Merck P56003). The pulse sequence used incorporated a double echo excitation sculpting component for water suppression (Bruker library zgesgp) to remove residual water. Samples were run using 256 scans with a relaxation delay D1 of 1 s. All spectra were phased, base corrected and calibrated for the pyrazine peak at 8.63975 ppm. The chemical shift of the diagnostic groups of SMM and DMSP-amine were the CH2-group adjacent to the sulphur (peak 3 in Fig. 3F). The initial rates obtained were then plotted against [S]o using QtiPlot to obtain the enzymatic parameters Kmand Vmaxas shown in Fig. 3e.

[0339] Sequence identification of homologs in other species

[0340] The S. anglica MMT, SDC and DOX protein sequences were used in a tBLASTn search against the selected species. For each species, the top hit was taken, then codon optimised for expression in E. coli. Sequences were then synthesised by commercial DNA synthesis and cloned into the Golden Gate modified pMALc2x plasmid. For SpSDC, which lacks genetic resources, DNA was extracted from S. patens material from Foz de Odeleite, Portugal. Primers designed against the full-length CDS of SaSDC were used to amplify the S. patens DNA by polymerase chain reaction (PCR) using high fidelity Phusion® polymerase (New England Biolabs M0530). Bands of equivalent size to S. anglica SDC were extracted and cloned into the pJET1 ,2 / blunt cloning vector (Thermo Scientific K1231). E.coli strain DH5a was transformed with this construct, and clones were selected and sequenced by Sanger sequencing using forward and reverse pJetl .2 sequencing primers. Sequences were then aligned against SaSDC and a full-length homolog was identified. This S. patens SDC homolog was cloned into the Golden Gate modified pMALc2x plasmid, as with other homologs.

[0341] RT-qPCR

[0342] RNA from S. anglica was extracted as described above. RNA samples were treated with TURBO™ DNAse (ThermoFisher Scientific, AM2238) according to the manufacturer’s instructions. Samples were then tested for contaminating DNA by PCR using RT-qPCR SaActin primers. Complementary DNA (cDNA) first strand synthesis was performed using Superscript IV® Reverse transcriptase (Thermo Fisher) according to manufacture protocol. Housekeeper genes SaActin and SaGAPDH were identified in the S. anglica transcriptome by BLAST. Specific primers pairs were designed using NCBI Primer BLAST specifying PCR product sizes of 120-200 bp and annealing temperatures of 60°C. RT-qPCR used total leaf RNA extracted from three independent biological replicates and was performed with three technical replicates, in 96-well plates using the RealTime PCR System (Aria system, Aglient). Primers showing 90-1 10% efficiency were used and amplification was performed using a programme consisting of 94°C for 2 minutes, followed by 40 cycles of 94°C for 15 s and 60°C for 1 minute, using SYBR® Green Jumpstart™ Taq ReadyMix™ (Merck S5193) according to the manufacturer’s protocol. For analysis, the Ct value of the target gene was normalised and expression values relative to each control were then determined by the Pfaffl method59.

[0343] Quantification of amino acids by Liquid Chromatography-Mass Spectrometry (LC-MS)

[0344] Leaf tissue from S. anglica was homogenised under liquid nitrogen and freeze dried. Amino acids were extracted from 30 mg of this powder as in60. Extracted samples were then derivatised with AccQ Tag (Waters) and diluted 1 :10-1 :1000 to bring in line with standard curves. Measurements were then performed as in60.

[0345] Differential expression analyses

[0346] Tomato and S. anglica mRNA libraries were constructed and sequenced with paired-ends by Novogene Co. Adapters were removed from reads using Trim Galore55. The tomato experiments were performed in quadruplet, and two of these biological replicates were then taken forward for RNA sequencing. For the S. anglica experiments, material from three independent biological replicates was used for each group (i.e. the three plants with the highest or lowest DMSP accumulation). Clean reads were then aligned either to the reference tomato transcriptome iTAG v4.1 for tomato sequencing or to our own trinity transcriptome constructed from S. anglica RNA-seq using HISAT261and default parameters. Aligned reads were then quantified using Salmon62run in alignment-based mode. Statistical testing of quantified reads was then performed using DESeq263. All gene ontology enrichments were performed using ShinyGO v0.7764.

[0347] Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES)

[0348] Plant samples were homogenised in liquid nitrogen, freeze dried and then digested in nitric acid. Soil samples were freeze dried and 1 ml volume of powder was incubated in 5 ml of 50 mM ammonium EDTA (Merck 330507) with shaking for 1 hour at room temperature. Samples were then centrifuged at 4000g for 10 minutes and filtered through a 0.22 pm filter. Plant digests and soil solutions were then analysed for elemental composition using an Agilent Vista Pro at the UEA Science Analytical Facility.

[0349] Nitrogen measurements

[0350] Plant and soil samples were homogenised at room temperature and freeze dried. 1.5 mg material was then weighed into nickel sleeves and assayed using Exeter CE440 CHN analyser calibrated using acetanilide and benzoic acid standards.

[0351] Construction of phylogenetic trees

[0352] BLASTp was run using SaMMTI , SaSDC and SaDOX as input sequences. The top 50 hits were taken along with homologous sequences from S. officinarum, S. lycopersicum and M. truncatula. SaCAOl and SaDAPDC were included to root the DOX and SDC trees, respectively. For SaMMT, trees were rooted against mmtN from Novosphingobium55. Rooting branches were then removed for visual clarity. Sequences were aligned using MUSCLE66and the subsequent Phylip file was run through PhyIML67with a standard bootstrap analysis of 100 repeats. The most similar hit for each species was retained and lower similarity accessions were removed so that each species was represented only once on the trees.

[0353] Quantification of MMT SDC DOX abundance in RNA-seq datasets

[0354] Primary RNA-seq data were collected for S. anglica, S. lycopersicum, A. thaliana, H. vulgare and N. benthamiana from the sources indicated in table S8. Adapters were trimmed as before and clean reads were aligned using Salmon62to de novo assembled transcriptome for S. anglica, iTAG v4.1 for S. lycopersicum, TAIR10 cDNA for A. thaliana, BaRT2v18 for H. vulgare and NBv5 for N. benthamiana. Mapping parameters were kept the same for all species. Homologs were identified for genes of interest using BLAST and reads mapping to them were then taken from each of the quantitative alignment files. Finally, for each library, the total number of reads mapping to each gene were divided to the total number of reads mapping to the ACT8 homolog to remove artefacts that may be introduced due to differences in sequencing depth and transcriptomic complexity. Plant materials and growth conditions

[0355] Tomato (S. lycopersicum cv. 'Micro Tom') seeds were obtained from Moles Seeds (www.wholesale.molesseeds.co.uk) and sterilised using 70% ethanol for 2 minutes, followed by 1 % sodium hypochlorite (Merck) for 20 minutes and rinsed five times with sterile water prior to sowing. Tomato growth media was prepared (Murashige and Skoog basal medium (Merck M5519), 3% sucrose, 0.8% agar) and five seeds were sown per 40 ml of media in a Magenta GA-7 Plant Culture Box (Bioworld) using sterile technique. Material was placed in dark conditions at 4°C for five days before transfer to a Sanyo MLR-351 controlled environmental chamber, at 26°C / 18°C, day / night with a 16-hour photoperiod, for2 weeks of growth. Plantlets at the 3-4 true leaf stage were transferred to treatment conditions using 40 ml fresh media, three plantlets per GA-7 box. Tomato growth medium was supplemented with ± 5 pM DMSP or 100 mM NaCI ± 5 pM DMSP. Phenotypes were observed after 2 weeks of growth under treatment conditions. Fresh weight was recorded for aerial and roots tissues per box. DMSP levels were measured for leaf. For each condition, four biological replicates of three technical replicates were used. For RNA-seq, two boxes were selected per condition.

[0356] For A. thaliana experiments, seedlings were sterilised in 1 % sodium hypochlorite for 13 minutes and then rinsed five times with sterile water. Seedlings were sown on % strength MS containing 0.25% glucose, stratified at 4°C for two days, and then grown in a Sanyo MLR-351 controlled environmental chamber, at 22°C / 18°C, day / night with a 16-hour photoperiod, for 12 days of growth. Plants were then transferred to treatment plates containing fresh % MS with 0.25% glucose ± 100mM NaCI and grown for a further 12 days before being harvested for further analyses.

[0357] For glasshouse experiments, Spartina anglica plants were collected from Stiffkey saltmarsh between May and July 2022 and maintained in freshwater until January 2024. Clonal clumps from six plants were each divided into three, and each sub-divided clump was then treated with either deionised water, sea salts (35 PSU) or NaCI (500 mM) for up to 5 weeks. The salinity of treatments with sea salts or NaCI was measured by electrical conductivity and maintained at 45-55 mS / cm throughout the experiment. Three leaf samples from each clump were pooled together and collected 1 , 3 and 5 weeks after treatment.

[0358] For tomato, barley, pea and wheat experiments, seeds of the indicated genotypes were germinated, and the plants were grown in standard compost under controlled environment or glasshouse conditions as indicated. Plants were subsequently treated with solutions of DMSP either by root drench (watering of soil) or spraying of foliar material using the concentrations and frequencies indicated.

[0359] Plasmid construction and creation of transgenic plants

[0360] Plasmids over-expressing MMT (pOsUBI3-SaMMT), SDC (p35S-SaSDC) and / or DOX (pAtUB / 10- DOX) were constructed using Golden Gate cloning, as previously described68. Transformation of N. benthamiana using Agrobacterium tumefaciens was performed as previously described68. Transgenic A thaliana plants were created by floral dip. Transgenic plants were selected on media containing kanamycin and all subsequent phenotypic analyses were performed on T3 material. For transgenic barley plants, a plasmid over-expressing MMT (pOsActinl-SaMMT), SDC (pOsUBI3- SaSDC) and DOX (pZmUB / 1-D0X) was constructed using Golden Gate cloning and Agrobacterium- mediated transformation was performed.

[0361] Quantification of DMSP lyase and demethylation gene abundance in metagenomic datasets

[0362] Raw data for S. alterniflora (accession numbers: SRR11828899, SRR11829104, SRR11829000) and S. patens (accession numbers: SRR11829102, SRR1 1828889, SRR1 1828593)73were retrieved from the NCBI Sequence Read Archive (SRA) using the SRA toolkit v.2.10.2. Data quality was assessed using FastQC vO.12.1 , and as the quality was deemed high, no trimming was necessary. Assembly (MEGAHIT v1.2.9), annotation (Prodigal v2.6.3), and non-redundant protein sequence generation (CD-HIT v4.8.1) were performed. Profile Hidden Markov Model (HMM)-based searches were conducted for the nine DMSP lyase genes (dddD, dddK, dddL, dddP, dddQ, dddU, dddW, dddX and dddY) and the DMSP demethylation gene (dmdA) using HMMER v.3.3 with an e-value threshold of 1 e-30. Relative abundance calculations were based on ten single copy marker genes77, retrieved with an e-value threshold of >1 e-10. Sequences were further refined by BLASTp (>40% identity and >70% coverage) to create a reference database. Metagenomic reads from the six samples were mapped to this database using CoverM 0.6.1. Relative abundances of ddd and dmdA genes were normalised to the average relative abundance of ten conserved single copy marker genes and total read counts as previously described78.

[0363] Example 5 Transgenic plant using bacterial / algal genes for DMSP synthesis

[0364] Marine algae, bacteria and corals are major global DMSP producers and utilise ‘methylation’ ‘transamination’ or ‘decarboxylation’ pathways for DMSP production (Figure 18). Enzymes involved in the DMSP synthesis pathways are described in Table 2.

[0365] Table 2 Enzymes involved in the DMSP synthesis pathways

[0366]

[0367]

[0368] As described above, these genes can be utilised to make transgenic plants with increased concentrations of DMSP.

[0369] Nucleic acid constructs containing one or more genes to reconstitute a DMSP synthesis pathway are created using standard cloning methods, e.g. traditional restriction enzymes or multi-gene construct assembly via Golden Gate cloning. The genes are expressed under the control of a promoter (e.g. a strong constitutive promoter such as pOsUBI3, p35S or pAtUBHO), and can be codon-optimised (e.g. using DNA synthesis) and / or include a synthetic intron to improve gene expression levels. The nucleic acid constructs reconstituting a DMSP synthesis pathway are then transformed into plants (e.g. using Agrobacterium-mediated transformation) to create transgenic plants that produce elevated concentrations of DMSP.

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[0443] Listing of Sequences

[0444] Candidate: SDC

[0445] Enzyme: Ornithine decarboxylase

[0446] Interpro ID: IPR008286

[0447] Name in text: SaODC (from Spartina anglica)

[0448] SEQ ID NO: 1

[0449] ATGGTTGGAGGCAGCCCCATGCCCATGCAGGCCATGCTGGTGGTCCCCGGCGTCAAGGACC GGAAGGTGCTCTCGTTCAAGGGCGACGCGCTCAAGGAGAACGACGCCATCCGCTCCATCAC CTCCTCCACCCCCGCCCAGCGTAGCGCCTTCCACGTCCTGGACCTGGCCAAGGTCGTCGAC CTGTACCGCGCGTGGCGGCGCGCGCTGCCCAACGTGCGCCCCTACTACGCCGTCAAGTGCA ACAGTGAACCGGCGCTGCTGGGCGCGCTGGCTGCGCTAGGCGCGGGCTTCGACTGCGCCA GCCACGCCGAGATCGACGCCGTGCTTGCGCTCGGCGTGCACCCGAGCAGCATCGTCTACGC

[0450] CAACCCGTGCAAGCCAGAGTCCCACCTCGAGCACGCCGCGGCGGCGGGCGTCAATCTCACC

[0451] ACATACGACTCCGAGGAGGAGGTGGCCAAGGTGCGGCGCTGCCACCCGAGGTGCGAGCTCC

[0452] TGCTCCGTATCAAGGGCCCCGACGACGAGGAGGCCCAGTGCGACCTGGGCATCAAGTACGG

[0453] CGCGCTCGCCGACGAGGTCGTGCCCCTCCTTCGCGCCGCGCAGCGCGCGGGGGTCCCCGT

[0454] CGCCGGCGTCGCGTTCAACGTTGGTAGCCCCGCGTCGCGCATGGACTTGTACCGCGACGCC

[0455] ATCGAGGCCGCGCGCGCGGCCTTCGACGCCGCGGCGGGCCTCGGCATGCCGCCCATGCGC

[0456] GTGCTCGACATCGGCGGCGGGTTCACGGCGGGTGCCACGTTCGAAGAGGCTGCCGCGGTGA

[0457] TCAATGACGCCCTCGCGCGGCACTTCGGCGACCTGCCGTACGTGGAGGTGATCAGCGAGCC

[0458] CGGGCGCTACTTCGTCAAGAAGGCCATCACGCTCGCCGCCCGTGTCATCGGGAAGCGCACG

[0459] CGCGGGGAGGTGCGTGAGTACTGGATCGACGACGGCGTCTACGGCTCGTTCAACTGCGTCA

[0460] TCATGGACCACTACAAGCCCCGCCCGAGGCCGCTGGTGACCCCGCGCCCCGGCGAGGCGAA

[0461] GTACACGTCGACGGTGTTCGGACCCACGTGCGACTCGCAAGACATGGTGGTCACCGGTTACA

[0462] AGCTGCCGGAAATGAGTGTCGGCGACTGGCTCGTCTTCGACGACATGGGTGCCTACACCACC

[0463] GCAGCCGGCTCCAAGTTCAACGGCTTTGACACCTCGGACATAAAAATCTACCTGGCATACTCC

[0464] ACCTGA

[0465] Candidate: SDC

[0466] Enzyme: Ornithine decarboxylase

[0467] Interpro ID: IPR008286

[0468] Name in text: SaODC (from Spartina anglica)

[0469] SEQ ID NO: 2

[0470] MVGGSPMPMQAMLVVPGVKDRKVLSFKGDALKENDAIRSITSSTPAQRSAFHVLDLAKVVDLYRA

[0471] WRRALPNVRPYYAVKCNSEPALLGALAALGAGFDCASHAEIDAVLALGVHPSSIVYANPCKPESHL

[0472] EHAAAAGVNLTTYDSEEEVAKVRRCHPRCELLLRIKGPDDEEAQCDLGIKYGALADEVVPLLRAAQ

[0473] RAGVPVAGVAFNVGSPASRMDLYRDAIEAARAAFDAAAGLGMPPMRVLDIGGGFTAGATFEEAA

[0474] AVINDALARHFGDLPYVEVISEPGRYFVKKAITLAARVIGKRTRGEVREYWIDDGVYGSFNCVIMDH

[0475] YKPRPRPLVTPRPGEAKYTSTVFGPTCDSQDMVVTGYKLPEMSVGDWLVFDDMGAYTTAAGSKF NGFDTSDIKIYLAYST

[0476] Candidate: DOX

[0477] Enzyme: Copper amine oxidase

[0478] Interpro ID: IPR000269

[0479] Name in text: SaDOX (from Spartina anglica)

[0480] SEQ ID NO: 3 AAGCCATTCGCCAAATCACCCCACTCCCTCAGCGACCACCGCGTCCCCTCCTCGGCTGCTTC

[0481] CGGCGGACGGAGAGAGAACCCCGGTCCCCTTCCTAGGGTTCCCGGAGCCCTAGCCCGAACC

[0482] CCGCACTCTCGAGGACCCCGGCGACTAGCAGGAGCGGCGGCGGAGAGGGAGGCAGAAGGC

[0483] GCGGGAGAGGCGGAGGGCCTGTGCTGTATGTCAGATCGTCTGTGCCGGCGAGGCGGAGGC

[0484] GGAGGAGGAGAATACGGACACGGTCAGCTCGCGAATCCGCTCCAATGGCCTCAGCTAAGGA

[0485] AAAGGCGGCGGTCTGCTGCGTCGGAGGCGCGCCCGCACGTGGCGAACCCGTTGCCGTGCG

[0486] GGCGATCCCGGAGTCCCCGCCCGGGAAGGTGGAGATGGCCGCGAGCGATGAACACGTGGC

[0487] CGCGTCGGCCGTCGGAGGTGGCGTCGTGATCGAGGAGATCGCCACGGTCCAGCCCACCATC

[0488] GCGAAGGCCTCTTCCAAAGGGATACCTATCATGACCAGAGCCCAGAGAAGCCACCCTTTAGA

[0489] CCCATTATCTGCTACTGAAATTGCAGTCGCTGTAGCAACTGTAAGGGCTGCTGGAAGGTCTCC

[0490] TGAGGAGAGAGACAGCATGCGTTTTTCCGAAGTTGTGCTGTTGGAACCGGAAAAGAATGTTGT

[0491] GGCATTAGCAGATGCGTACTTTTTCCCACCTTTCCAACCATCACTGCTTCCTAGGTCCAAAGGT

[0492] TCTGCTGTTATTCCAAGCAGGTTACCTCCTAGGAGGGCCAAGCTTGTCATTTACAATAAGCATT

[0493] CAAATGAGACTAGCATTTGGATAGTGGAACTCTCTGAAGTGCATGCAGCTACTAGGGGTGGAC

[0494] ATCACAGAGGCAAGGTGATATCGTCTGAAGTTATTCCAGATGTTCAGCCTGCAATGGATGCTA

[0495] TGGAGTATGCTGAGTGCGAAGCAACAGTCAAAAGCTATCCCCCATTTATTGAAGCTATGAAGC

[0496] GAAGAGGTGTGGATGACATGGATCTTGTTATGGTAGATACCTGGTGTGCTGGCTACTATGGTG

[0497] ATGCTGATGCTCCCAGTCGCAGACTTGGCAAACCTCTAATATTTTGCAGAACTGAGAGTGATA

[0498] GCCCCATGGAGAATGGTTATGCACGTCCTGTGGAAGGGATTCATGCCATTGTTGATGTACAGA

[0499] ATAATACTGTGATTGAATTTGAAGATAGGAAGTTGGTTCCTCTCCCCCCACCAGATCATTTGAG

[0500] AAACTATACTCCTGGGGAAACAAGAGGTGGAGTTGATCGAAGTGATGTAAAACCGCTTATTAT

[0501] CAATCAACCTGAAGGTCCAAGTTTTCGCATTAATGGCTATTTTGTGGAGTGGCAGAAGTGGAA

[0502] CTTCCGTATTGGCTTCACCCCCAAAGAGGGTTTGGTTATCCATTCTGTTGCGTACGTTGATGG

[0503] TAGCCGTGGACGTCGACCTATAGCACATAGGCTGAGCTTTGTTGAGATGGTTGTTCCTTATGG

[0504] AGATCCAAATGAACCACATTATAGTAAGAATGCATTTGATGCTGGAGAAGATGGACTTGGAAAA

[0505] AATGCCAATTCTCTTAAGAAGGGGTGTGACTGCTTGGGCTACATAAAATATTTTGATGCACATT

[0506] TCACAAACTTCACTGGTGGTGTGGAGACAATTGAGAACTGTGTCTGTTTGCATGAGGAGGACC

[0507] ATGGAATCCTGTGGAAACATCAAGACTGGAGAACTGGCTTGGCAGAAGTTAGGCGGTCAAGG

[0508] AGGCTCACTGTCTCATTTATCTGTACAGTTGCAAACTATGAATATGGTTTTTACTGGCACTTTCA

[0509] TCAGGATGGCAACATAGAAGCAGAAGTAAAGCTAACTGGAATTCTCAGCTTGGGAGCTCTAAT

[0510] GCCTGGGGAATCTAGGAAATATGGCACAACCATTTCTCCTGCTCTGTATGCACCAGTTCATCA

[0511] GCACTTTTTTGTTGCTCGTATGGACATGGCTATTGATTGCAAACCTAACGAGGCTTATAATCAG

[0512] GTGGTTGAGGTGAACACTAAAGTGGAAAGTCCTGGACCACATAATGTGCATAACAATGCTTTC

[0513] TATGCTGAAGAGAAGTTGTTGAAGTCTGAGTTGCAAGCTATGCGAGACTGTGACCCGTCATCT

[0514] GCACGCCATTGGATTGTAAGGAACACAAGAACTGTAAATCGAACAGGACAGCCAACTGGTTAC

[0515] AAGCTCGTGCCGGGTTCAAACTGTTTGCCGTTGGCTCTGCCAGAGGCAAAATATCTCAGGAG

[0516] AGCTGGGTTCTTAAAGCATAATCTCTGGATCACGCAGTACAAAAGTGACCAGAATTTCCCGGG GGGAGAATTTCCAAATCAGAACCCACGTCTTCATGAGGGCTTACCTACATGGGTGAAGAAGGA

[0517] TAGATCCCTGGAGGAAACTGACATTGTCCTCTGGTACGTGTTCGGGCTCACCCATATCCCAAG

[0518] GCTGGAAGACTGGCCTGTGATGCCGGTGGAGCGTATTGGCTTCATGCTCATGCCACATGGAT

[0519] TCTTCAACTGCTCCCCTGCGGTGGACGTGCCTCCCAGCTCTTCTGATGCCGATGCCAAGGAG

[0520] GCCGAGTCGCCCAAGGCCATGCAGAACGGCCTTCTGTCGAAGCTGTGAGCTCCGCTGTTCCT

[0521] CGGACGAACAACTGGCTTGCTGCTGTTTCGACTTTTATCTTACAACGCTGTTCCAATAAAAGG

[0522] GAAACTGTTGTCCCCTCTCTTGTACTGTTCCAATAAAAGCGATACTGTGTTCTCCGCTCCCTTG

[0523] TGATTCGTATGGTGATCTGTAGTTGCTGTTCTTGTTGATCAGCGAGCTATGAACTATCACATAG

[0524] TATAATAGCACATAATAATGTTTGGGCATGCATCTACATCGTTAAAATCATGTGCTATAGTGTG

[0525] GCGATCCCTTTCTGCTCTCTGCCTTTTTTTTCCAATGTCGAGGGCCGCTACAAACAACAAATCG

[0526] AGACCACCTGGTTCCTCGACAATAAAAATTTGATCAAGAAGCAATGTCTCGAAACTTTAACAAT

[0527] TAACACAATGCGTACTTGCTGG

[0528] Candidate: DOX

[0529] Enzyme: Copper amine oxidase

[0530] Interpro ID: IPR000269

[0531] Name in text: SaDOX (from Spartina anglica)

[0532] SEQ ID NO: 4

[0533] MASAKEKAAVCCVGGAPARGEPVAVRAIPESPPGKVEMAASDEHVAASAVGGGWIEEIATVQPTI

[0534] AKASSKGIPIMTRAQRSHPLDPLSATEIAVAVATVRAAGRSPEERDSMRFSEVVLLEPEKNWALA

[0535] DAYFFPPFQPSLLPRSKGSAVIPSRLPPRRAKLVIYNKHSNETSIWIVELSEVHAATRGGHHRGKVI

[0536] SSEVIPDVQPAMDAMEYAECEATVKSYPPFIEAMKRRGVDDMDLVMVDTWCAGYYGDADAPSR

[0537] RLGKPLIFCRTESDSPMENGYARPVEGIHAIVDVQNNTVIEFEDRKLVPLPPPDHLRNYTPGETRG

[0538] GVDRSDVKPLIINQPEGPSFRINGYFVEWQKWNFRIGFTPKEGLVIHSVAYVDGSRGRRPIAHRLS

[0539] FVEMVVPYGDPNEPHYSKNAFDAGEDGLGKNANSLKKGCDCLGYIKYFDAHFTNFTGGVETIENC

[0540] VCLHEEDHGILWKHQDWRTGLAEVRRSRRLTVSFICTVANYEYGFYWHFHQDGNIEAEVKLTGIL

[0541] SLGALMPGESRKYGTTISPALYAPVHQHFFVARMDMAIDCKPNEAYNQVVEVNTKVESPGPHNVH

[0542] NNAFYAEEKLLKSELQAMRDCDPSSARHWIVRNTRTVNRTGQPTGYKLVPGSNCLPLALPEAKYL

[0543] RRAGFLKHNLWITQYKSDQNFPGGEFPNQNPRLHEGLPTWVKKDRSLEETDIVLWYVFGLTHIPR

[0544] LEDWPVMPVERIGFMLMPHGFFNCSPAVDVPPSSSDADAKEAESPKAMQNGLLSKL

[0545] Candidate: MMT

[0546] Enzyme: Methionine S-methyltransferase

[0547] Interpro ID: IPR025779

[0548] Name in text: SaMMTI (from Spartina anglica) SEQ ID NO: 5

[0549] GCCACTCGAACCAACCTCCGATCTCCTGAACAACCAACCTCCGATCTCCTAAACCAATGGCGG

[0550] GCGAGGACAAGGACGTCGATGCCTTCCTCGCCGACTGCACCGTCTCCGGCGACGCCGCCTA

[0551] CAACGCCGCCAAGGCCGTGCTCGAGCGCCTCCACGCTCCGGCCACCAGGCCCGCCGCGCG

[0552] ACGCCTCCTCGGCGCTGTACGCCGACGCTTTGCCCGAGACCCCGCCACCGGGGAGGACTGC

[0553] TTCCGCACCTACCACTTCCGCATCCACGACGTCCAGCTTGACCCTCACATCCAAGGTTTCCAA

[0554] CAAAGAAAGAAGCTAACAATGATGGACATACCTAGCATTTTCATTCCTGAAGATTGGTCCTTCA

[0555] CTTTCTATGAGGGCCTCAATCGGCATCAAGACTCCATTTTCAGGGATAAGACTGTAGCCGAGC

[0556] TGGGATGTGGCAATGGTTGGATATCCATTGCTCTTGCAGAAAAGTGGTCCCCTTCAAAGGTCT

[0557] ACGGTCTTGATATAAACCCAAGAGCTGTGAAGATTGCGTGGATAAACCTGTACCTAAATGCAC

[0558] TAGATGATGATGGTCTCCCAATCTATGACAAGGAGGGGAAAACATTGCTGGACAGAGTTGAAT

[0559] TCCATGAATCAGATCTCCTTTCTTATTGCAGAGACAACAACATGGAGCTCGATCGCATAGTTGG

[0560] ATGCATACCACAGATTCTTAACCCAAATCCAGAGGCAATGTCAAAGATTATAACTGAAAATTCA

[0561] AGCGAGGAGTTCTTGTACTCCTTGAGTAACTATTGCGCTCTTCAGGGTTTTGTTGAGGATCAAT

[0562] TTGGCCTTGGATTGATCGCTCGGGCAGTTGAAGAAGGTATTGCTGTCATAAAGCCTTCAGGTA

[0563] TTATGGTATTCAACATGGGAGGTCGGCCAGGACAAGGTGTCTGTGAACGTTTATTTGAGCGCC

[0564] GAGGATTTCGCATAACAAAGCTCTGGCAAACAAAAATTATGCAGGCTGCTGACACAGATATTT

[0565] CAGCTTTAGTTGAAATTGAGAAAAATAGCCGACATCGCTTTGAGTTCTTCATGGATCTTGTTGG

[0566] GGATCAGCCTATATGTGCTCGCACAGCCTGGGCATATATGAAATCTGGTGGCCGCATTTCACA

[0567] TGCTCTATCTGTGTATAGCTGTCAACTTCGCCAACCCAACCAGGCTAAGAAAATATTCGAGTTT

[0568] CTTAGAGATGGATTCCGTGAAGTCAGCAGTTCTCTTGATTTATCCTTTGACGATGATTCTGTTG

[0569] CTGATGAAAAAATTCCGTTCCTTGCGTACCTAGCTAGCTTTTTGAAAGAGAATAAGTCTAATCC

[0570] CTGTGAGCCACCAGCTGGATGCCAAAACTTCCGGAAACTTGTTGCTGGATTTATGAAGAGCTA

[0571] TCATCACATCCCGTTAGCTCCTGATAATGTCGTTGTGTTCCCTTCTCGCGCTGTGGCAATAGA

[0572] GAATGCTCTTCAGTTGTTCTCACCCGTGCTAGCAATTGTTGATGAGCATTTGACCAGACACTTG

[0573] CCCAAACACTGGTTAACATCTTTAGCAATTGAGGGGGGAGCAGATGGTAACCATGCTGAAGAC

[0574] ACAGTCACTGTAATTGAGGCACCACGCCAATCAGATTTGTTGATTGAATTAATCAAGAAGCTGA

[0575] AGCCTCAAGTTGTTGTTACGGGCATGGCTCAATTTGAGGCTATCACTAGTGCTGCTTTTGAGA

[0576] ACTTACTGAGCGTAACGAAAGATGTTGGTTCCCGGCTATTCCTAGACATTTCAGAGCATTTGG

[0577] AATTGTCTAGTCTACCAAGCTCTAATGGTGTGTTGAAATATCTTGCTGGAAAGACTCTACCTTC

[0578] GCATGCTGCTATTCTGTGTGGTTTAGTAAAGAACCAGGTTTATTCTGATCTGGAAGTTGCTTTT

[0579] GCCATATCTGAGGATGCATCTGTATACAAAGCTTTGTCACAGACTATTGAGCTAATGGAGGGG

[0580] CATACTTCTATGATCAGTCAACACTATTATGGTTGCCTTTTCCATGAGCTTCTCGCATTTCAAAT

[0581] TGTTGACCGACATCCACAACAAGAGAGAGAACCTGCAGAAGTTATACCTCAGAAGATGATTGG

[0582] ATTTTCTAATTCGGCCATGTCTACACTAAAGGCAGCTGAATTTTTCGTTCCTGATTCAAATGAAT

[0583] CCAGTATCATTCATATGGATTTAGACCGCAGCTTTCTGCCAGTACCTTCTGCAGTGAACGGTTC

[0584] TGTTTTTGAAAGTTTTGTCAGGCAGAACATCACTGAATCTGAAACTGATGTTCGTTCCAGCATT CAACAGCTGGTGAAAGATAGTTATGGTTTCCCTGCAGATGCCTGTTCCGAATTTATCTATGGCA

[0585] ACACCTCTCTTGCACTGTTCAACAAGCTTGTTCTTTGTTGCATTCAAGAAGAGGGCACCATGCT

[0586] TTTCCCCTTGGGCACTAATGGCCGTTACATTTCTGCAGCAAAGTTTGTGAACGCAAAAACCTTA

[0587] ACTATTCCAACATCACTCAGTTCAGATTTCAAGATTGAACCGAAAGCTCTAGCTGACACTCTTA

[0588] AGAATGTATCTCGTCCATGGGTATATATTTGTGGCCCCACAATCAATCCTACCGGTTTTCTGTA

[0589] CAGTGACAATGATATTCAGGATCTGCTCTCTGTATGTGCTGAATATGGTGCTAGGGTAGTGTTA

[0590] GATACCTCCTTCTCTGGTCTGGAGTTCCAAACTGATGGCTGGAGTCGGTGGAATTTGGAAAGA

[0591] TCTCTTTCTGCTGTGAACTGTTCAAAGGCTTCATTTTCCGTGCTTCTGCTTGGAGAGCTGTCCT

[0592] TTGAGCTAACTGCAGCGGGGCATGAATTTGGGTTTCTGATTCTGAATGACTCATCCTTGATCG

[0593] AGACATTTTACAGTTTCCCGAGCTTGAGTCGGCCGCACAGCACATTGAAGTACACTTTCAAAA

[0594] GGCTACTAGGCCTTAAGAACCAGAAGGATGAGCACTTCTCTAATCTAATGGTGGAGCAAATGG

[0595] AGAAATTAAAGAGCCGCGCCAACCACTTGATAAAGACACTTGAGAGCTGCGGCTGGGACGTT

[0596] GCCAGTGGTTGTGGTGGCACCTCAATGCTGGCAAAACCGACCGCATACATTGGGAAGCCCTT

[0597] CAAAGTTGATGGGTTTGAGGGCAAGCTGGATAGCTGCAACATCAGGGAGGCCATGCTGAGAG

[0598] CCACCGGCCTGTGCATAAACAGCAGCTCGTGGACCGGGATCCCGGACTACTGCCGATTCAGC

[0599] TTTGCTGTGGGGAGCAGCGAATTCGAACGCGCCACCGGTTGCATAACTCGCTTCAAGGAGTT

[0600] GGTTCTGGAGTAGCTGCTCAGATCGCCCCCAATATTTAAATTAGTAACAGCAGCAACAAACGC

[0601] TTGCTGTGAATGTATTGGTAACACGCTCATTGGCTCTTGTGACTAGAGTCTATTCATCACTGGC

[0602] GAATCAATCAGTCAAGTGCAACTTTGAATAAACACTTCCTGTTTGCATCATTATAGCTCCAAATC

[0603] AATATTTTGCAAATTGGCCTACCGTTGACTACTTGACTTTGAAATGCAAGCACTTTAAATATGCC

[0604] GCCCTATCGCGGACCCATCACGTGATAGGGCAAACGTGTGACCCGCTCGCCTTCGTCTTTGC

[0605] AGTGCTGCCCCGCCTCTTGTCTCGCTCGTCCTTCCTCTGCCTCACCACTTGTCGTGCCCTTGT

[0606] CACCAAGCCGATCCTCCACCTCCACCTGGATCAC

[0607] Candidate: MMT

[0608] Enzyme: Methionine S-methyltransferase

[0609] Interpro ID: IPR025779

[0610] Name in text: SaMMTI (from Spartina anglica)

[0611] SEQ ID NO: 6

[0612] MAGEDKDVDAFLADCTVSGDAAYNAAKAVLERLHAPATRPAARRLLGAVRRRFARDPATGEDCF

[0613] RTYHFRIHDVQLDPHIQGFQQRKKLTMMDIPSIFIPEDWSFTFYEGLNRHQDSIFRDKTVAELGCGN

[0614] GWISIALAEKWSPSKVYGLDINPRAVKIAWINLYLNALDDDGLPIYDKEGKTLLDRVEFHESDLLSYC

[0615] RDNNMELDRIVGCIPQILNPNPEAMSKIITENSSEEFLYSLSNYCALQGFVEDQFGLGLIARAVEEGI

[0616] AVIKPSGIMVFNMGGRPGQGVCERLFERRGFRITKLWQTKIMQAADTDISALVEIEKNSRHRFEFF

[0617] MDLVGDQPICARTAWAYMKSGGRISHALSVYSCQLRQPNQAKKIFEFLRDGFREVSSSLDLSFDD

[0618] DSVADEKIPFLAYLASFLKENKSNPCEPPAGCQNFRKLVAGFMKSYHHIPLAPDNVVVFPSRAVAIE

[0619] NALQLFSPVLAIVDEHLTRHLPKHWLTSLAIEGGADGNHAEDTVTVIEAPRQSDLLIELIKKLKPQW VTGMAQFEAITSAAFENLLSVTKDVGSRLFLDISEHLELSSLPSSNGVLKYLAGKTLPSHAAILCGLV

[0620] KNQVYSDLEVAFAISEDASVYKALSQTIELMEGHTSMISQHYYGCLFHELLAFQIVDRHPQQEREP

[0621] AEVIPQKMIGFSNSAMSTLKAAEFFVPDSNESSIIHMDLDRSFLPVPSAVNGSVFESFVRQNITESE

[0622] TDVRSSIQQLVKDSYGFPADACSEFIYGNTSLALFNKLVLCCIQEEGTMLFPLGTNGRYISAAKFVN

[0623] AKTLTIPTSLSSDFKIEPKALADTLKNVSRPWVYICGPTINPTGFLYSDNDIQDLLSVCAEYGARVVL

[0624] DTSFSGLEFQTDGWSRWNLERSLSAVNCSKASFSVLLLGELSFELTAAGHEFGFLILNDSSLIETFY

[0625] SFPSLSRPHSTLKYTFKRLLGLKNQKDEHFSNLMVEQMEKLKSRANHLIKTLESCGWDVASGCGG

[0626] TSMLAKPTAYIGKPFKVDGFEGKLDSCNIREAMLRATGLCINSSSWTGIPDYCRFSFAVGSSEFER

[0627] ATGCITRFKELVLE

[0628] SEQ ID NO: 7 MMT from Zea mays

[0629] >ZmMMT_nucleotide_sequence

[0630] ATGGGCAGCGTGGGCGTGGAGGCGGATGACGACGGCAGCATCGTCAAGGAGTTCCTGCAGC

[0631] GGTGCGAGCCCTCCGGCGACGCCGCCTACGGCGAGCTCAGGGCTCTGCTGGCGCGCCTCC

[0632] ACGACCCGGCCACCAGGCGCGACGCGCGCCTCTTCCTCGCCGCGCTCCACCGCCACCAGCA

[0633] GCGCAGCTCGTCGGCCGCCGGCGGACTAACGCACGAGCAATTCTTTCGACGCTTCGGCTTCC

[0634] GCATGCAGGAGCTGCTGCTCCAGGATACCGCCACCGACATAACCGCCTCCAGCTTCATCGCC

[0635] ACCAAGCCCGCCGCAGCCGCAGGTTTCCAACAAATGAAGAAGTTGACAATGATGGAGATACC

[0636] AAGCATCTTCATTCCTGAAGATTGGTCATTCACTTTCTATGAGGGCCTCAACCGTCATCCAGAC

[0637] TCCATTTTCAGGGATAAGACAGTAGCTGAGCTGGGATGTGGCAACGGTTGGATATCTATTGCT

[0638] CTTGCAGAGAAGTGGTGCCCTTCAAAGGTCTATGGCCTGGATATAAACCCAAGAGCTGTGAAG

[0639] ATTGCATGGATAAACCTGTACTTGAACGCATTAGATGACGACGGTCTCCCGATCTATGACGGG

[0640] GAGGGGAAAACATTGCTCGATAGAGTTGAATTCTATGAATCTGATCTCCTTTCTTACTGTAGAG

[0641] ACAACAAGATAGAGCTCGATCGTATTGTTGGATGCATACCACAGATTCTCAACCCAAATCCAG

[0642] AGGCGATGTCAAAGATTGTAACAGAGAATTCAAGTGAGGAGTTCTTGTACGCCTTGAGTAACT

[0643] ACTGTGCTCTTCAGGGTTTTGTCGAGGACCAATTTGGCCTTGGGTTGATTGCTCGTGCGGTGG

[0644] AAGAAGGGATATCTGTCATAAAGCCTTCAGGTATTATGGTATTCAACATGGGAGGTCGACCAG

[0645] GACAGGGTGTCTGTGAACGTCTATTTCGACGGCGTGGGTTTCGCATTACTAAGCTCTGGCAAA

[0646] CCAAAATTATGCAGGCTGCTGACACAGATATTTCAGCTTTGGTTGAAATTGAGAAAAACAGCAG

[0647] ACATCGATTCGAGTTCTTCATGGATCTTGTTGGGGATCAGCCTATCTGTGCTCGCACAGCCTG

[0648] GGCATACATGAAATCTGGTGGTCACATTTCACATGCTTTGTCTGTGTATAGCTGTCAACTTCGC

[0649] CAGCCCAACCAGGTGAAGAAAATATTTGAGTTTCTTAAAGATGGATTCCATGAAGTCAGCAGTT

[0650] CCCTTGATTTATCCTTTGACGATGATTCTGTAGCTGAGGAAAAAATTCCCTTCCTAGCTTACCT

[0651] TGCTAGTTTTCTGAAAGAGAACAAGTCAAATCCCTGTGAGCCACCAGCTGGATGTCTAAACTTT

[0652] AGGAAACTTGTTGCTGGATTTATGAAGAGCTACCATCACATTCCTCTAACTCCTGATAATGTCG

[0653] TTGTGTTCCCTTCTCGCTCTGTGGCAATAGAGAATGCTCTTCAATTGTTCTCACCGGCGCTTGC

[0654] AATTGTTGATGAACATTTAACCAGACACTTGCCCAAGCAATGGTTAACATCTTTAGCAATTGGA

[0655] AGAGCAGATTGTAATCACGCCGATGGTACAGTTACTGTAATTGAGGCACCACGCCAATCAGAT TTACTGATTGAGTTGATCAGGAAGCTGCAGCCTCAGGTGGTTGTTACTGGCATGGCTCAATTT

[0656] GAGGCTATCACCAGTGCTGCTTTTGAGAACCTACTAAACGTAACAAAAGATGTTGGCTCCCGG

[0657] CTGTTCCTGGATATTTCTGAGCATCTGGAGTTGTCTAGCCTGCCAAGCTCTAATGGTGTATTGA

[0658] AATATCTTGCTGGAAAGACATTACCGTCACATGCAGCTATACTGTGTGGTTTAGTAAAGAATCA

[0659] GGTGTACTCTGATCTGGAGGTTGCTTTTGCCATTTCCGAAGATGCAGCTGTATATAAAGCATTA

[0660] TCACAAACTATTGAGCTATTGGAAGGCCACACTTCTCTGATCAGCCAGCACTATTATGGTTGCC

[0661] TTTTCCACGAGCTTCTGGCATTTCAGATTGCTGACCGTCATCCACAGCAAGAGAGACAACCTG

[0662] CGGAAGTGATACCTCAGCAGATGATAGGATTTTCTGACCCAGCTGTGTCTACCCTAAAGGCTA

[0663] CTGAATTTTTCGTTCCTGGTTCAGCTGAATCCAGCATTATTCATATGGATTTAGATCGCAGCTT

[0664] CCTGCCAGTACCTTCAGCAGTGAATGCGTCTGTTTTTGAAAGTTTTGTCAGGCAGAACATCACT

[0665] GATTCTGAAACCGATGTCCGTTCCAGCATCCAACAGCTGGTGAAAGACAGCTATGGTTTATCC

[0666] GCAGCTGGTTGTGCAGAAATTATCTACGGAAACACCTCCGTAGCACTCTTCAACAAGCTCGTC

[0667] CTTTGTTGCATGCAAGAACAGGGCACCTTGCTTTTCCCCTTGGGCACCAATGGCCACTACGTC

[0668] TCCGCAGCAAAGTTTGTGAACGCTAGCACCGTGACTATACCGACAAATCCCAGTTCAGGGTTC

[0669] AGGATCGAGCCCAAGGTTCTAGCTGACACTCTTAAGAATGTATCTCGGCCGTGGGTGTATGTT

[0670] TGCGGCCCCACGATCAACCCTACTGGCTTTCTGTACAGTGACAGTGACATCCGAGAGCTGCT

[0671] CTCCGTATGCGCTGAATATGGAGCCAGGGTAGTGATAGATACATCCTTCTCTGGCCTGGAGTA

[0672] CGAGACCGATGGATGGAGGCAGTGGAATCTGGCAGGATGCCTTTCCTCTTTGAAGCGTTCAG

[0673] AGCCATCGTTCTCTGTGGTCCTGCTCGGAGAGCTGTCCTTTGCGCTGACTGCAGGTGGGCAT

[0674] GACTTTGGGTTTGTGATATTGGGTGACTCGTCGTTGGCTGAGACGTTCCATAGTTTTTCTAGCT

[0675] TGAGTCGACCGCACACCACATTGAAGTACACTTTCAAGAAGCTGCTGGGGCTTAAGAACCAGA

[0676] AGGACCAGCATTTCTCGGATCTGATAGTGGAGCAGAAGGAGGAGCTGAAGAATCGCGCCAAC

[0677] CAGTTGATACAGACACTGGAGAGCTGTGGCTGGGAAGCTGCCATCGGTTGCGGTGGCATCTC

[0678] GATGCTGGCGAAGCCCACCGCGTACATGGGGAAGGCCTTCAAAGCTGCTGGCTTCGACGGC

[0679] GAGCTGGACGCGAGCAACATCCGGGAAGCCATCCTGAGGGCCACTGGGCTGTGCATAAACA

[0680] GCAGCTCCTGGACGGGGATCCCTGGCTACTGCCGGTTCAGCTTTGCTCTGGAGAGGGGGGA

[0681] ATTCGAGCGTGCGATGGGATGCATAGCTCGGTTCAAGGAGCTGGTTCTGGGAGGCGCTCAGA

[0682] TGAATGGTGCCTGA

[0683] SEQ ID NO: 8 MMT from Zea mays

[0684] >Zm M MT_prote i n_seq u e n ce

[0685] MGSVGVEADDDGSIVKEFLQRCEPSGDAAYGELRALLARLHDPATRRDARLFLAALHRHQQRSS

[0686] SAAGGLTHEQFFRRFGFRMQELLLQDTATDITASSFIATKPAAAAGFQQMKKLTMMEIPSIFIPEDW

[0687] SFTFYEGLNRHPDSIFRDKTVAELGCGNGWISIALAEKWCPSKVYGLDINPRAVKIAWINLYLNALD

[0688] DDGLPIYDGEGKTLLDRVEFYESDLLSYCRDNKIELDRIVGCIPQILNPNPEAMSKIVTENSSEEFLY

[0689] ALSNYCALQGFVEDQFGLGLIARAVEEGISVIKPSGIMVFNMGGRPGQGVCERLFRRRGFRITKLW

[0690] QTKIMQAADTDISALVEIEKNSRHRFEFFMDLVGDQPICARTAWAYMKSGGHISHALSVYSCQLRQ

[0691] PNQVKKIFEFLKDGFHEVSSSLDLSFDDDSVAEEKIPFLAYLASFLKENKSNPCEPPAGCLNFRKLV AGFMKSYHHIPLTPDNVVVFPSRSVAIENALQLFSPALAIVDEHLTRHLPKQWLTSLAIGRADCNHA

[0692] DGTVTVIEAPRQSDLLIELIRKLQPQVWTGMAQFEAITSAAFENLLNVTKDVGSRLFLDISEHLELS

[0693] SLPSSNGVLKYLAGKTLPSHAAILCGLVKNQVYSDLEVAFAISEDAAVYKALSQTIELLEGHTSLISQ

[0694] HYYGCLFHELLAFQIADRHPQQERQPAEVIPQQMIGFSDPAVSTLKATEFFVPGSAESSIIHMDLDR

[0695] SFLPVPSAVNASVFESFVRQNITDSETDVRSSIQQLVKDSYGLSAAGCAEIIYGNTSVALFNKLVLC

[0696] CMQEQGTLLFPLGTNGHYVSAAKFVNASTVTIPTNPSSGFRIEPKVLADTLKNVSRPWVYVCGPTI

[0697] NPTGFLYSDSDIRELLSVCAEYGARVVIDTSFSGLEYETDGWRQWNLAGCLSSLKRSEPSFSVVLL

[0698] GELSFALTAGGHDFGFVILGDSSLAETFHSFSSLSRPHTTLKYTFKKLLGLKNQKDQHFSDLIVEQK

[0699] EELKNRANQLIQTLESCGWEAAIGCGGISMLAKPTAYMGKAFKAAGFDGELDASNIREAILRATGL

[0700] CINSSSWTGIPGYCRFSFALERGEFERAMGCIARFKELVLGGAQMNGA

[0701] SEQ ID NO: 9 Alternate transcript present in in S. anglica: SaMMT2

[0702] >SaMMT2_nucleotide_sequence

[0703] AATCAGCTAAAGCTAAAGCCACCTATCTCTCGGTTGTTAATAATAAGCTGAAGAAGGAAATGG

[0704] GCAGCGAGGGCGTGGTGGTGGATTTCCTGAAGCGGTGCGAGCCCTCCGGCGACGCCGCCTA

[0705] CGGGGAGCTCAAGGCACTGCTCGGCCGCCTCCATGACCCAGCAACCAGGCGCCAAGCTCGC

[0706] GTCTTCCTGGCCGCGCTCCGGGGCCATTCTTCCGATGATGGCGGTGACTTGTTCTTCCGGCG

[0707] GTACGGCTTCGCCATCCGGGAGCTGGACCTTCGTCAATCCGTCCCCTTCTTCCATCATGCAG

[0708] GAGCGTCATCAGCAGGTTTCCAACAAAGAAAGAAGCTAACAATGATGGACATACCTAGCATTT

[0709] TCATTCCTGAAGATTGGTCCTTCACTTTCTATGAGGGCCTCAATCGGCATCAAGACTCCATTTT

[0710] CAGGGATAAGACTGTAGCCGAGCTGGGATGTGGCAATGGTTGGATATCCATTGCTCTTGCAG

[0711] AAAAGTGGTCCCCTTCAAAGGTCTACGGTCTTGATATAAACCCAAGAGCTGTGAAGATTGCGT

[0712] GGATAAACCTGTACCTAAATGCACTAGATGATGATGGTCTCCCAATCTATGACAAGGAGGGGA

[0713] AAACATTGCTGGACAGAGTTGAATTCCATGAATCAGATCTCCTTTCTTATTGCAGAGACAACAA

[0714] CATGGAGCTCGATCGCATAGTTGGATGCATACCACAGATTCTTAACCCAAATCCAGAGGCAAT

[0715] GTCAAAGATTATAACTGAAAATTCAAGCGAGGAGTTCTTGTACTCCTTGAGTAACTATTGCGCT

[0716] CTTCAGGGTTTTGTTGAGGATCAATTTGGCCTTGGATTGATCGCTCGGGCAGTTGAAGAAGGT

[0717] ATTGCTGTCATAAAGCCTTCAGGTATTATGGTATTCAACATGGGAGGTCGGCCAGGACAAGGT

[0718] GTCTGTGAACGTTTATTTGAGCGCCGAGGATTTCGCATAACAAAGCTCTGGCAAACAAAAATTA

[0719] TGCAGGCTGCTGACACAGATATTTCAGCTTTAGTTGAAATTGAGAAAAATAGCCGACATCGCTT

[0720] TGAGTTCTTCATGGATCTTGTTGGGGATCAGCCTATATGTGCTCGCACAGCCTGGGCATATAT

[0721] GAAATCTGGTGGCCGCATTTCACATGCTCTATCTGTGTATAGCTGTCAACTTCGCCAACCCAA

[0722] CCAGGCTAAGAAAATATTCGAGTTTCTTAGAGATGGATTCCGTGAAGTCAGCAGTTCTCTTGAT

[0723] TTATCCTTTGACGATGATTCTGTTGCTGATGAAAAAATTCCGTTCCTTGCGTACCTAGCTAGCT

[0724] TTTTGAAAGAGAATAAGTCTAATCCCTGTGAGCCACCAGCTGGATGCCAAAACTTCCGGAAAC

[0725] TTGTTGCTGGATTTATGAAGAGCTATCATCACATCCCGTTAGCTCCTGATAATGTCGTTGTGTT

[0726] CCCTTCTCGCGCTGTGGCAATAGAGAATGCTCTTCAGTTGTTCTCACCCGTGCTAGCAATTGT

[0727] TGATGAGCATTTGACCAGACACTTGCCCAAACACTGGTTAACATCTTTAGCAATTGAGGGGGG AGCAGATGGTAACCATGCTGAAGACACAGTCACTGTAATTGAGGCACCACGCCAATCAGATTT

[0728] GTTGATTGAATTAATCAAGAAGCTGAAGCCTCAAGTTGTTGTTACGGGCATGGCTCAATTTGAG

[0729] GCTATCACTAGTGCTGCTTTTGAGAACTTACTGAGCGTAACGAAAGATGTTGGTTCCCGGCTA

[0730] TTCCTAGACATTTCAGAGCATTTGGAATTGTCTAGTCTACCAAGCTCTAATGGTGTGTTGAAAT

[0731] ATCTTGCTGGAAAGACTCTACCTTCGCATGCTGCTATTCTGTGTGGTTTAGTAAAGAACCAGGT

[0732] TTATTCTGATCTGGAAGTTGCTTTTGCCATATCTGAGGATGCATCTGTATACAAAGCTTTGTCA

[0733] CAGACTATTGAGCTAATGGAGGGGCATACTTCTATGATCAGTCAACACTATTATGGTTGCCTTT

[0734] TCCATGAGCTTCTCGCATTTCAAATTGTTGACCGACATCCACAACAAGAGAGAGAACCTGCAG

[0735] AAGTTATACCTCAGAAGATGATTGGATTTTCTAATTCGGCCATGTCTACACTAAAGGCAGCTGA

[0736] ATTTTTCGTTCCTGATTCAAATGAATCCAGTATCATTCATATGGATTTAGACCGCAGCTTTCTGC

[0737] CAGTACCTTCTGCAGTGAACGGTTCTGTTTTTGAAAGTTTTGTCAGGCAGAACATCACTGAATC

[0738] TGAAACTGATGTTCGTTCCAGCATTCAACAGCTGGTGAAAGATAGTTATGGTTTCCCTGCAGAT

[0739] GCCTGTTCCGAATTTATCTATGGCAACACCTCTCTTGCACTGTTCAACAAGCTTGTTCTTTGTT

[0740] GCATTCAAGAAGAGGGCACCATGCTTTTCCCCTTGGGCACTAATGGCCGTTACATTTCTGCAG

[0741] CAAAGTTTGTGAACGCAAAAACCTTAACTATTCCAACATCACTCAGTTCAGATTTCAAGATTGA

[0742] ACCGAAAGCTCTAGCTGACACTCTTAAGAATGTATCTCGTCCATGGGTATATATTTGTGGCCCC

[0743] ACAATCAATCCTACCGGTTTTCTGTACAGTGACAATGATATTCAGGATCTGCTCTCTGTATGTG

[0744] CTGAATATGGTGCTAGGGTAGTGTTAGATACCTCCTTCTCTGGTCTGGAGTTCCAAACTGATG

[0745] GCTGGAGTCGGTGGAATTTGGAAAGATCTCTTTCTGCTGTGAACTGTTCAAAGGCTTCATTTTC

[0746] CGTGCTTCTGCTTGGAGAGCTGTCCTTTGAGCTAACTGCAGCGGGGCATGAATTTGGGTTTCT

[0747] GATTCTGAATGACTCATCCTTGATCGAGACATTTTACAGTTTCCCGAGCTTGAGTCGGCCGCA

[0748] CAGCACATTGAAGTACACTTTCAAAAGGCTACTAGGCCTTAAGAACCAGAAGGATGAGCACTT

[0749] CTCTAATCTAATGGTGGAGCAAATGGAGAAATTAAAGAGCCGCGCCAACCACTTGATAAAGAC

[0750] ACTTGAGAGCTGCGGCTGGGACGTTGCCAGTGGTTGTGGTGGCACCTCAATGCTGGCAAAAC

[0751] CGACCGCATACATTGGGAAGCCCTTCAAAGTTGATGGGTTTGAGGGCAAGCTGGATAGCTGC

[0752] AACATCAGGGAGGCCATGCTGAGAGCCACCGGCCTGTGCATAAACAGCAGCTCGTGGACCG

[0753] GGATCCCGGACTACTGCCGATTCAGCTTTGCTGTGGGGAGCAGCGAATTCGAACGCGCCACC

[0754] GGTTGCATAACTCGCTTCAAGGAGTTGGTTCTGGAGTAGCTGCTCAGATCGCCCCCAATATTT

[0755] AAATTAGTAACAGCAGCAACAAACGCTTGCTGTGAATGTATTGGTAACACGCTCATTGGCTCTT

[0756] GTGACTAGAGTCTATTCATCACTGGCGAATCAATCAGTCAAGTGCAACTTTGAATAAACACTTC

[0757] CTGTTTGCATCATTATAGCTCCAAATCAATATTTTGCAAATTGGCCTACCGTTGACTACTTGACT

[0758] TTGAAATGCAAGCACTTTAAATATGCCGCCCTATCGCGGACCCATCACGTGATAGGGCAAACG

[0759] TGTGACCCGCTCGCCTTCGTCTTTGCAGTGCTGCCCCGCCTCTTGTCTCGCTCGTCCTTCCTC

[0760] TGCCTCACCACTTGTCGTGCCCTTGTCACCAAGCCGATCCTCCACCTCCACCTGGATCAC

[0761] SEQ ID NO: 10 >SaMMT2_protein_sequence

[0762] MGSEGVVVDFLKRCEPSGDAAYGELKALLGRLHDPATRRQARVFLAALRGHSSDDGGDLFFRRY

[0763] GFAIRELDLRQSVPFFHHAGASSAGFQQRKKLTMMDIPSIFIPEDWSFTFYEGLNRHQDSIFRDKTV AELGCGNGWISIALAEKWSPSKVYGLDINPRAVKIAWINLYLNALDDDGLPIYDKEGKTLLDRVEFH

[0764] ESDLLSYCRDNNMELDRIVGCIPQILNPNPEAMSKIITENSSEEFLYSLSNYCALQGFVEDQFGLGLI

[0765] ARAVEEGIAVIKPSGIMVFNMGGRPGQGVCERLFERRGFRITKLWQTKIMQAADTDISALVEIEKNS

[0766] RHRFEFFMDLVGDQPICARTAWAYMKSGGRISHALSVYSCQLRQPNQAKKIFEFLRDGFREVSSS

[0767] LDLSFDDDSVADEKIPFLAYLASFLKENKSNPCEPPAGCQNFRKLVAGFMKSYHHIPLAPDNVVVF

[0768] PSRAVAIENALQLFSPVLAIVDEHLTRHLPKHWLTSLAIEGGADGNHAEDTVTVIEAPRQSDLLIELIK

[0769] KLKPQVVVTGMAQFEAITSAAFENLLSVTKDVGSRLFLDISEHLELSSLPSSNGVLKYLAGKTLPSH

[0770] AAILCGLVKNQVYSDLEVAFAISEDASVYKALSQTIELMEGHTSMISQHYYGCLFHELLAFQIVDRH

[0771] PQQEREPAEVIPQKMIGFSNSAMSTLKAAEFFVPDSNESSIIHMDLDRSFLPVPSAVNGSVFESFV

[0772] RQNITESETDVRSSIQQLVKDSYGFPADACSEFIYGNTSLALFNKLVLCCIQEEGTMLFPLGTNGRY

[0773] ISAAKFVNAKTLTIPTSLSSDFKIEPKALADTLKNVSRPWVYICGPTINPTGFLYSDNDIQDLLSVCAE

[0774] YGARVVLDTSFSGLEFQTDGWSRWNLERSLSAVNCSKASFSVLLLGELSFELTAAGHEFGFLILND

[0775] SSLIETFYSFPSLSRPHSTLKYTFKRLLGLKNQKDEHFSNLMVEQMEKLKSRANHLIKTLESCGWD

[0776] VASGCGGTSMLAKPTAYIGKPFKVDGFEGKLDSCNIREAMLRATGLCINSSSWTGIPDYCRFSFAV

[0777] GSSEFERATGCITRFKELVLE

[0778] SEQ ID NO: 11 Burl >Burl|WP_009894054.1 diaminopimelate decarboxylase [Burkholderia thailandensis]

[0779] MNANPDALKLTRAHAAPPADWWARERLHYRDNSLHFCGRRVAELAAAFAEPVFLYDPQRAVDNV

[0780] ARLQRALGDLQRGDFHVYYAMKANRFRPLLSELRRSPIFGIDACSPEELREALACGFAPERISYTA

[0781] HGMMPDEAALLAALPDVHVNCDTLSAIALLGSRSPGREIGIRVNPGVGIGYGDSERLSYAGATITKF

[0782] GIYAEQFGAALELAARHGLTVTWLHCHAGCGYLDAQLDSFERVLDALDAFVARAPGLRGINLGGG

[0783] LGLPHRATDRPLDLERWRAAVHARFGGRPLALAIEPGDFIAKDAGMLVLRVAYVELKRNRRFVGL

[0784] NGGFNLAIEPAFYDLPCEPVPCVRRPGPTQSVCLAGNINEALDVWGDDVSLPPVEPGDFVALLNA

[0785] GGYASSMSSNHCLRGQFRELALFDAAPH

[0786] SEQ ID NO: 12 BurD >BurD|WP_009894048.1 aminotransferase [Burkholderia thailandensis]

[0787] MAHRNDSLLTSAQQDARHLLHPWADLTALGRETPTVIVDAQGTRVTDAEGRTYLDAIGGMWCVT

[0788] VGYGRREIADAIRDQALRMPFYTPFGAMTNAPAAALGARLAERAPGDLKRVHLTTCGSTAVESAL

[0789] RFAHYYFGATGRPHKRHIVTRGDAYHGSTYLAASVSGKAWDRTCFHYDSTIVHHLSSPNPYRRPA

[0790] GMSVAAFCASLVDEFDALIAKLGADRIACFIAEPILASGGVIVPPPGYLAAMRERCRRHDILYISDEV

[0791] VTGFGRVGHFFASQAHFGIEPDMIVVAKGLTSGYQPLGAVLISERLVEAVSGERAYGNGVFTNGFT

[0792] YSGHPVACAAALANIELMERERICEHVRDVGPYFIRRLDALRRFPIVGDVRGDHLMACIECTSGAG

[0793] ATGALPTPADIAIAQRVDRHCEEMGLLVRPYESMCILSPPLTVTRADIDEICEILAAALERTQRELAE

[0794] RAPVREENETC

[0795] SEQ ID NO: 13 DsyGD >DsyGD|WP_044616208.1 [Gynuella sunshinyii YC6258] MKQVSYEISSQVLEQYDSPQGRAFYRQVMGDSGFNIHYGIYPSENETMKTASENIIRHLQELAQQ

[0796] RGVHLPQASILDLGSGTGGAAHYLAGHFGCHVTCVNISPEQNKINRKQAQELGIDDLIKIEQCSFDN

[0797] LPGKWSGQFDLVWSEEAFCHAEHKDTVIKEAWRVLKPGGVLVFSDIMEGELNQDTHTFSDRNAIR

[0798] DLASPSDYIRLCMANGFYHLSYHDLSHHLPINFRKMIDQIDQHYDRLVDNGVSSKYADNFRQSLND

[0799] RVNAAFQGNFSWGSFVMNKSTRLEHPHLRSVIEGRNLCRITAEPLTRENLAELGTLYAYDQPLEQ

[0800] HPPVPQHSWPVKYPQRMETGRGLAPLGTDDMTMTWQQEILHARNEVLEHCYQNIAYRDEQHGY

[0801] FIEWFNQHVESGQKFYCPGVPLLYVLAAPVDHPKAQDFKAFIADGSHGVIINPGVWHTNPIPLIDTE VTLTTTQSIVDASCDCSLSAEHNQWLNITVSTGTDS

[0802] SEQ ID NO: 14 DSYE >DSYE|MMETSP0113 [Norrisiella sphaerica BC52]

[0803] SEVGSKVVEQYDNLQTRKFYQTVMGGGGNDIHFGVFKSKNDNVREAAANTTELMLDLMHWIRPV

[0804] TQHHQVLDLGSGHGGATHEMIKRFGTQNTCFNIGTEQNEMNLAYCQEIGISDHVTCHRGDFNLGL

[0805] PKEWDEGFDYVYSCEVLCHAADKPALLSEVRRVLAPGGVFVFSDIMGADGVSEEILKPFTDRNTT

[0806] TVMGRPSEYHYQLREAGLREVAYIDLTVNLGPFFQLMLDQVTNNYTELVEVGLSNDYLDNWISSLT DRVKVQRDHGAFAWGVFGAVNDQN

[0807] SEQ ID NO: 15 DsyB >DsyB_prok|AOR83342.1 methyltransferase [Labrenzia aggregata LZB033]

[0808] MPVARSLETAEEISDIAFGFMGSKALFSALHVDLFSLLSEKTLTPQHVAEESELDLDRATTLLTALTS

[0809] LGLVRREGAGFTNSPAAEAFLVKGRKYDFGDYLRFQIDKQMYPFMTQLNDALTDSLEDDQVASYE

[0810] TWFSDPEEARLYSRSQHAGSLGPGRGLAKLVDLSAAKQLLDVGGGTGAFSISLCKAYPGLRSTVL

[0811] DFPNVAKVGEEFIAEEGLQDRIRYAPGNALKDTWPDSADAVLMSYLFSGVPGTAIPGLVRKAFEVL

[0812] TPGSDFMVHDFMVDENRDGPKLAALWQLQHTAFNPEARSITSSYVAGLMEAAGFTDIAVEVMIPG MTMLVHGRKPE

[0813] SEQ ID NO: 16 DSYB >DSYB_euk|Prymnesium_parvum_CCAP_946 / 6

[0814] MLRLAPRLPTRALVRHALRAHALPLARPALPSGTRLFASAPADDIDVDNVAYGFMASQALFTGLEM

[0815] GLFDAIAAGPEAGLNIDELKAAANCSAPRLQTLVTSLVAIKSLKRTPDGRYTLSPNTARFLVQSSKQ

[0816] YYGDYLKYQMGRQFYHRMGALPDVMTSGEAPSYASWFSDPETAATYTQAQHNGSVATAKYLIKK

[0817] KLQLGDAATMLDVGGGSGAFSYVFTEATPGLSSTVLELPEVCRTGEAIKAKQPVSVQERVKLVEL

[0818] DATSPDWPVNDAAYDWLMSYISGSVPESIIGALYANAYKALKPGGRLLVHDFMVDNSLDGPPLGA

[0819] LWALQHVTVNADGLGLCPQGVIERMGSAGFDPSACETMEMITGLTKLIVAYKP

[0820] SEQ ID NO: 17 TpMMT >TpMMT|XP_002291473.1 methyl transferase-like protein [Thalassiosira pseudonana CCMP1335]

[0821] MTDYMFDVATGLLGDDGGVGAEEKTIKYVDLGSGTGAAALRLCQKHDVIAKATCLNLCEEQNALA

[0822] RKCASDLGLEDRIAVVTGTYESAPFEANSFDIAFSQDAFVHAFSKVGTFREALRVTKPGGVLVFCD

[0823] LMCGSGDGVSEEELATFAATNMVNDWLSPDLNVRACQEAGWTDVKFVDLTLDIRISFQLMLKKVE

[0824] KIIQDGNPAKIDEKLLDSYKKNLANRIVQVDRGVFKWGWTGKKPLYKFCTRKDSW SEQ ID NO:18 MmtN >MmtN|GAM03459.1 S-adenosyl-l-methionine: l-methionine s- methyltransferase [Novosphingobium sp. MBES04] MSDADGSKVISRHDDPSATTERPGYAFDPTDPWTITFQQGLKAAGLEGKAVYEVGVGTGTNVAFV LRHCAAKVFYGSDLDPRLVELARRNVANLAPERADSFQPVEGAVSLIDTDEARAKIARTDVVIGCL

[0825] PQVGDPNDERFAAFRAEHAVDLPQGADDEAQDHIAHYYPWAMFDEYPYNSVGLGLNEALLRRIK

[0826] EQAPKADWMNFGCRIGSDLIFEMFRANGYEPEKLASQLVLQHAGTDISFFVTLEGALTGTDLEGE FVCRFFADPLGHEPLSARAAQALLDKDPNVPLYHEVAVIRGTPKMD

[0827] SEQ ID NO:19 BurB >BurB|WP_009894046.1 SET domain-containing protein-lysine N- methyltransferase [Burkholderia thailandensis]

[0828] MTITLIEPVQQRTGIYPSSDLKVEDGYPSSDTFQIIQTQDGRGAGVRVLKTFARGRRMARVSGQITA

[0829] FCRLHTLQINAHTHLYDPHFSGLLLHSCDPNVRLDMAGFELWSLRDIAAGEMLTMDYASTEDVLM RQFECHCGAPNCRRWITGAKELPNDIGQALLAGLRAAALA

[0830] SEQ ID NO: 20 Burl >bur / |NC_007650.1 :2573204-2574466 Burkholderia thailandensis E264 chromosome II, complete sequence ATGAACGCGAATCCCGACGCACTCAAGCTCACGCGCGCGCACGCCGCGCCGCCCGCCGACT

[0831] GGTGGGCGCGCGAGCGGCTGCACTACCGCGACAACTCACTGCACTTCTGCGGCCGCCGCGT

[0832] CGCCGAACTCGCGGCCGCGTTCGCCGAACCGGTGTTCCTGTACGACCCGCAGCGCGCGGTC

[0833] GACAACGTCGCGCGCCTGCAACGCGCGCTGGGCGACCTGCAACGCGGCGACTTTCACGTCT

[0834] ACTACGCGATGAAGGCGAATCGCTTCCGCCCGCTGCTGTCCGAGCTGCGGCGCTCGCCGAT

[0835] CTTCGGCATCGACGCGTGCTCGCCCGAGGAGTTGCGCGAAGCGCTCGCGTGCGGCTTCGCG

[0836] CCGGAGCGCATCTCGTACACCGCGCACGGCATGATGCCCGACGAAGCCGCGCTCCTCGCCG

[0837] CGCTGCCCGACGTGCACGTGAACTGCGACACGCTGAGCGCGATCGCGCTGCTCGGCAGCCG

[0838] TTCGCCGGGGCGCGAGATCGGCATTCGCGTGAACCCCGGCGTCGGCATCGGCTACGGCGAC

[0839] AGCGAGCGCCTGAGCTACGCGGGCGCGACCATCACGAAGTTCGGCATCTATGCGGAACAGT

[0840] TCGGCGCGGCGCTCGAGCTCGCCGCGCGGCACGGCCTCACCGTGACGTGGCTGCATTGCCA

[0841] CGCGGGCTGCGGCTATCTCGACGCGCAGCTCGACTCGTTCGAGCGCGTGCTCGATGCGCTC

[0842] GATGCGTTCGTCGCGCGCGCGCCGGGGCTGCGCGGCATCAACCTCGGCGGCGGGCTCGGC

[0843] CTGCCGCATCGCGCGACCGACCGCCCGCTCGATCTCGAACGCTGGCGCGCGGCCGTGCAC

[0844] GCGCGCTTCGGCGGCCGGCCGCTCGCGCTCGCGATCGAGCCAGGCGACTTCATCGCGAAG

[0845] GACGCCGGCATGCTCGTGCTGCGCGTCGCATACGTCGAGCTCAAGCGCAACCGCCGTTTCG

[0846] TCGGATTGAACGGCGGCTTCAATCTCGCGATAGAACCCGCGTTCTACGATCTGCCGTGCGAG

[0847] CCCGTGCCGTGCGTGCGCCGGCCGGGCCCCACGCAATCGGTGTGCCTCGCGGGCAACATCA

[0848] ACGAAGCGCTCGACGTGTGGGGCGACGACGTGAGCCTGCCGCCCGTCGAGCCGGGCGATTT

[0849] CGTCGCGCTGCTGAACGCGGGCGGCTACGCGTCATCGATGAGCTCGAACCACTGCCTGCGC GGGCAGTTCCGCGAGCTCGCGCTGTTCGACGCCGCGCCGCACTGA SEQ ID NO: 21 BurD >burD|NC_007650.1 :2555749-2557170 Burkholderia thailandensis E264 chromosome II, complete sequence

[0850] ATGGCACACCGAAACGATTCCCTTCTGACGAGCGCGCAGCAGGACGCGCGGCATCTGCTGC

[0851] ACCCGTGGGCCGACCTGACCGCGCTCGGCCGCGAGACGCCGACCGTCATCGTCGACGCGC

[0852] AAGGCACGCGCGTGACCGACGCCGAAGGCCGCACGTATCTCGACGCGATCGGCGGCATGTG

[0853] GTGCGTGACCGTCGGCTACGGCCGGCGCGAGATCGCGGACGCGATCCGCGACCAGGCGCT

[0854] GCGCATGCCGTTCTACACGCCGTTCGGCGCGATGACGAACGCGCCCGCCGCCGCGCTCGGC

[0855] GCGCGGCTCGCCGAGCGCGCGCCCGGCGATCTGAAGCGCGTGCATCTGACCACCTGCGGC

[0856] TCGACCGCGGTCGAATCGGCGCTGCGTTTCGCGCACTACTACTTCGGCGCGACGGGCCGCC

[0857] CGCACAAGCGCCACATCGTCACGCGCGGCGACGCGTACCACGGCAGCACGTATCTCGCCGC

[0858] GTCGGTGTCCGGCAAGGCGTGGGACCGCACGTGCTTTCACTACGACAGCACGATCGTCCATC

[0859] ACCTGTCGTCGCCGAACCCGTACCGGCGGCCGGCCGGCATGAGCGTCGCCGCGTTCTGCGC

[0860] GTCGCTCGTCGACGAATTCGACGCGCTGATCGCAAAGCTCGGCGCGGACCGGATCGCCTGC

[0861] TTCATCGCCGAGCCGATCCTCGCGTCGGGCGGCGTGATCGTGCCGCCGCCCGGTTATCTCG

[0862] CCGCGATGCGCGAGCGCTGCCGCCGGCACGACATCCTGTACATCTCCGACGAAGTCGTCAC

[0863] GGGCTTCGGCCGCGTCGGCCATTTCTTCGCGTCGCAGGCGCACTTCGGCATCGAGCCCGAC

[0864] ATGATCGTCGTCGCGAAGGGACTCACGTCCGGCTACCAGCCGCTCGGCGCGGTGCTGATCT

[0865] CGGAGCGGCTCGTCGAGGCGGTGTCCGGCGAGCGCGCGTACGGCAACGGCGTGTTCACGA

[0866] ACGGCTTCACCTATTCGGGCCATCCGGTCGCGTGCGCGGCGGCGCTCGCGAACATCGAGCT

[0867] GATGGAGCGCGAGCGGATCTGCGAGCACGTGCGCGACGTCGGCCCGTACTTCATTCGCCGG

[0868] CTCGACGCGCTGCGGCGCTTTCCCATCGTCGGCGACGTGCGCGGCGACCACCTGATGGCGT

[0869] GCATCGAATGCACGAGCGGCGCGGGCGCGACGGGCGCGCTACCGACGCCCGCGGACATCG

[0870] CGATCGCGCAGCGCGTCGATCGCCATTGCGAGGAGATGGGCCTGCTCGTGCGGCCCTACGA

[0871] GAGCATGTGCATCCTGTCGCCGCCCCTCACCGTGACCCGCGCCGACATCGATGAAATCTGCG

[0872] AGATTCTCGCGGCCGCGCTCGAGCGCACGCAGCGCGAGCTCGCGGAACGCGCCCCCGTTC GCGAGGAAAACGAAACATGCTGA

[0873] SEQ ID NO: 22 DsyGD >dsyGD|NZ_CP007142.1 :c1328831 -1327347 Gynuella sunshinyii YC6258 chromosome, complete genome

[0874] ATGAAACAAGTCAGTTATGAAATCAGCTCACAGGTTCTGGAACAATACGATTCACCACAAGGA

[0875] CGAGCGTTTTACCGGCAAGTAATGGGGGACAGCGGTTTCAATATCCATTACGGCATCTATCCT

[0876] TCCGAAAACGAAACCATGAAGACTGCCAGTGAAAATATCATTCGACATTTGCAGGAACTGGCG

[0877] CAACAACGTGGTGTTCACCTGCCCCAGGCATCCATACTGGATCTCGGCTCCGGTACCGGTGG

[0878] TGCCGCACATTATCTGGCCGGACATTTTGGTTGCCATGTTACCTGTGTGAACATCAGTCCGGA

[0879] ACAAAACAAAATTAATCGCAAACAGGCTCAGGAGCTGGGCATTGATGATTTGATCAAGATAGA

[0880] ACAATGCAGCTTCGACAATCTGCCCGGGAAGTGGTCAGGGCAGTTTGATCTGGTCTGGAGTG AAGAAGCCTTTTGCCACGCGGAGCATAAGGATACCGTGATCAAAGAAGCCTGGCGGGTGTTA AAACCCGGAGGCGTGCTGGTCTTCAGCGATATTATGGAGGGGGAGCTGAATCAGGACACCCA TACCTTCAGCGACCGTAACGCCATTCGCGATCTTGCCAGTCCAAGTGACTATATCCGGTTGTG CATGGCAAACGGTTTTTACCATTTAAGCTATCACGACCTCAGTCATCACCTGCCGATCAACTTC CGCAAAATGATCGATCAGATCGATCAACACTATGACCGACTGGTCGACAACGGAGTGTCATCC AAATATGCCGATAATTTCCGTCAATCATTGAACGACAGAGTCAATGCGGCTTTCCAGGGTAATT TTTCCTGGGGTAGTTTTGTGATGAACAAATCCACCCGGTTGGAACATCCGCATCTTCGCTCCG TGATCGAAGGGCGCAACCTCTGCCGTATCACAGCAGAACCACTGACCCGGGAAAATCTCGCC GAACTGGGAACCCTGTACGCCTATGATCAACCACTGGAACAACACCCACCGGTTCCACAACA CAGCTGGCCGGTAAAATATCCACAGCGAATGGAAACCGGGCGGGGTCTGGCTCCCCTTGGC ACCGATGATATGACCATGACATGGCAGCAGGAAATTCTGCATGCCCGTAATGAAGTGCTGGAA CATTGTTACCAGAACATCGCCTATCGGGATGAACAGCATGGATACTTTATCGAATGGTTCAAC CAGCACGTCGAAAGCGGACAAAAATTCTACTGTCCTGGTGTACCGTTGCTGTACGTGCTGGCA GCTCCGGTAGATCATCCCAAAGCGCAGGATTTTAAGGCATTTATCGCCGACGGCAGTCATGG CGTCATCATCAATCCCGGCGTATGGCACACCAATCCGATTCCACTGATCGATACAGAAGTTAC CCTGACCACTACTCAGTCTATTGTCGATGCCAGTTGTGACTGCTCACTGTCAGCAGAGCACAA TCAGTGGTTGAACATCACGGTCAGCACCGGGACGGACTCATGA

[0881] SEQ ID NO: 23 DSYE >CDS_DSYE|KV784358.1 :c1861226-1861136,c1860956- 1860739, c1860497-1859886 Fragilariopsis cylindrus CCMP1102 unplaced genomic scaffold FRACYscaffold_6, whole genome shotgun sequence ATGGCGCCTCCTACTGAAGAAGAAAACACAGGAGATTTCAAGGACCAACAATTCGGCGAGAA AGAGGTAGCTGATAAAGTTGTTGAGCAATATGACGAGAAACAAGCCAGAGTCTTTTACAAATA CGTCATGGGTGGCGGCGGATTTGATATTCATTATGGAATATTCCGTAAAGCAACAGATACTGT CTATGAATCATCAAAGGCGACCAATGAACGTTTATTGACTTGTCTCGATTGGACATTCCCTGTT ACTAAAGATTCATACGTTCTTGATCTTGGGTCTGGTCATGGTGGAATTTCACATGAGATTGCTG CCAGATTTGGTTGTAAGGTGATGGGAGCCAATATCTCTCCCGAACAAAATAAAATGAATTTGTT GGAGGCAAAGAAAGAAGGTGTTGGAGAACTGGTCGATGTTACGCTATGCAATTTCAACGATG GACTTCCAGAAGAATGGACCAATAAATTCACTCATGTTATTTCCTGTGAGGTTCTTTGTCACGC AGCAAACAAACCCGAACTATTCAAACAGATCAACCGTATCCTTCAACCGGGTGGTGCGTTTGT ATTTACTGATATTATGGGAGCAGATGGTGCAGATGAAAAAGTCTTGAAGGACTTCACTGATAGA AATGCGACTACTGCTATGGCCCGACCTAGTGAATATTTGACTCTTATTACTGATGCCGGACTTA GTGAACCATCATTTTTGAACTTTAGTCCACATTTGGTACATTACTTCCAATCGATGGTCAATCAG ATCAATGAAAATAAGGCCGCCATGATGAAGGAGGGTTGCTCAGAAGAATATCTAGCAAAATGG TTGGAATCGTTGACTAGTCGTGTTGATATGCAGCGTGAACACAATGTCTTTGCTTGGGGTATCT TCACGGCACGAACGGAGGGTAATATTTACTAA SEQ ID NO: 24 DsyB >dsyB_prok|KT989543.1 Labrenzia aggregata strain LZB033 DsyB (dsyB) gene, complete cds ATGCCCGTCGCCAGGTCCTTGGAAACGGCTGAGGAAATTTCAGATATTGCTTTCGGATTTATG GGCTCCAAGGCTCTTTTTTCTGCACTTCATGTAGATTTGTTTTCACTGCTTTCCGAAAAGACCT TGACGCCACAGCATGTTGCTGAAGAAAGCGAGCTTGATCTTGATCGCGCCACAACACTGTTGA CCGCGCTTACGAGCCTCGGGCTGGTGCGCCGGGAGGGAGCAGGCTTCACCAATTCGCCCGC AGCGGAGGCGTTTCTGGTGAAAGGACGGAAGTATGATTTCGGCGACTATCTGCGGTTTCAGA TCGACAAGCAGATGTACCCCTTCATGACGCAGCTCAATGACGCCCTGACGGACAGTCTGGAG GACGACCAGGTTGCTTCCTATGAGACCTGGTTCAGCGACCCGGAAGAAGCACGGCTTTATTC CCGGTCCCAGCATGCAGGCTCGCTTGGGCCCGGGCGGGGACTTGCCAAGCTTGTGGATCTG TCCGCTGCCAAGCAGTTGCTGGACGTTGGCGGTGGCACCGGGGCCTTCTCGATTTCGCTTTG CAAGGCGTATCCGGGACTGCGATCAACCGTTCTGGATTTTCCGAATGTCGCCAAGGTAGGTG AAGAGTTCATTGCCGAAGAAGGCCTGCAGGACCGCATCCGCTATGCGCCGGGCAATGCCCTG AAGGACACATGGCCGGACAGCGCGGACGCAGTGCTGATGTCCTATCTCTTTTCCGGCGTCCC GGGCACGGCCATTCCCGGGCTTGTGCGCAAGGCCTTCGAGGTTCTGACACCGGGCAGTGAC TTCATGGTGCATGACTTCATGGTCGACGAAAACCGCGACGGGCCGAAGCTGGCGGCACTCTG GCAGCTTCAGCACACCGCCTTCAACCCGGAAGCCCGTTCGATCACGAGTTCCTACGTTGCCG GCTTGATGGAAGCCGCCGGCTTCACAGATATCGCAGTTGAAGTGATGATCCCGGGCATGACG ATGCTGGTGCACGGCCGCAAGCCGGAATAG

[0882] SEQ ID NO: 25 DSYB >CDS_DSYB_euk|JWZX01001700.1 :c6450-5786,c5696-5210 Chrysochromulina tobinii strain CCMP291 CAF.Contig7645, whole genome shotgun sequence ATGCTGCTCGTCACTGCTCGGCGAGGCCTCTTCCAGTCGCGTGCAGTCTTGCGGGCGACGC CCGCCCTCGCGACGGCCGCGCCGGCGCTGATTGGCGCACGTCGCACTCTCTGCAACTCGCT GTTCGAGTTGTCCGACGACATTGACGTGGACAGCGTGTCATATGGCTTCATGGCATCGCAGG CGATCTTTTGCGCGCTCGAGGTGGGCGTATACGAGGCAGTTGCGGGCGCGGGCGAGGGCG GCCTTACGGCCGATGCCCTCAAGACTGCGTGCAGCATAAACGCGCCGCGGTTGCAGACGCT GCTCACGTCGCTCACTGCGATCAAGTCGCTCAAGCGCAACGCCGCCACGGGCGCTTACACG CTCTCGCCCAACAGCGCACGCTACCTTGTCCAGTCGTCGCGCCAATACTACGGCGACTACCT GCGCTACCAGATGGGTCGCCAGTTCTACCACCGCATGGGCGCGCTGCCAGATGTGATGACG ACCGGCGAGGCACCGTCATACGCGTCGTGGTTCTCGGACCCGGCCACCGCGGCGATGTACA CGCAGGCGCAGCATAACGGCTCGGTCGCCACGGCCAAGTACCTTGTCAAGAAAAAGCTTCAG CTCGGCTCGGTCACGTCAATGCTCGACGTGGGCGGCGGCTCGGGCGCCTTTTCGTACGTCTT CGTCGAGAGCACTCCGGGACTCAAGTCGACAGTCCTCGAGCTGCCTGAGGTGTGCAAGACG GGCAACGGCATCAAGGCGAAGCAGTCGCCCGATGTGCAGGCACGCGTCGCATTCATCGAGC TCGACGCGACGAAGCCCGACTGGCCGGTGAAGGAAGCGGACTATGAGCTTGTGCTCATGAG CTATATATCGGGTTCGGTGCCCGAGGCCATCATCGGCAAGCTGTACGAGAACGCGTTCAAGG CTCTCAAGCCGGGGGGTCGGCTGCTCGTGCACGATTTTATGGTGAACGACTCGCTCGACGGC

[0883] CCAGCGCTTGGTTCGCTCTGGGCGCTGCAGCACGTGACGGTCAACGCCGGCGGACTCGGTC

[0884] TGACCCCCGCGGAGGTCATCAGCCGCATGACCGCGGCGGGCTTTGCCAAGTGCGAGACGAT GGAGGTGATCAATGGCCTGACTAAGCTGATCGTCGCGTGCAAGGAGTGA

[0885] SEQ ID NO: 26 TpMMT >CDS_7pMMT_methyl transferase-like protein [Thalassiosira pseudonana CCMP1335]

[0886] ATGACGGATTATATGTTTGATGTTGCAACTGGATTGCTTGGTGATGATGGTGGAGTTGGAGCG

[0887] GAGGAGAAAACGATCAAGTATGTTGATTTGGGTAGTGGTACAGGAGCGGCTGCTTTGAGGTT

[0888] GTGTCAGAAACATGATGTCATCGCAAAGGCTACTTGTTTGAATCTGTGTGAGGAGCAGAATGC

[0889] TTTGGCAAGGAAGTGTGCCTCTGATCTTGGATTGGAAGACCGTATTGCCGTCGTGACAGGCA

[0890] CCTACGAATCAGCTCCGTTCGAAGCCAACTCGTTCGACATAGCCTTTAGCCAAGATGCATTCG

[0891] TCCATGCTTTCTCCAAGGTCGGAACATTCCGCGAGGCATTACGTGTTACCAAGCCTGGTGGAG

[0892] TCCTTGTCTTTTGCGACCTAATGTGCGGCAGCGGAGACGGGGTATCTGAAGAAGAGCTGGCA

[0893] ACATTTGCCGCGACCAACATGGTGAATGATTGGCTTTCTCCCGATCTGAATGTCAGGGCGTGT

[0894] CAGGAGGCTGGATGGACGGATGTCAAGTTTGTTGACTTGACGCTTGATATCCGTATCAGCTTT

[0895] CAACTCATGTTGAAGAAGGTGGAGAAGATCATTCAGGATGGAAATCCTGCGAAGATTGATGAA

[0896] AAGTTGTTGGATTCGTATAAGAAGAACTTGGCAAATAGGATTGTCCAGGTGGATAGGGGTGTG

[0897] TTCAAATGGGGTGTGGTGACTGGGAAGAAGCCGTTGTACAAGTTTTGCACCAGAAAGGATTCT TGGTAA

[0898] SEQ ID NO: 27 MmtN >mmf / V|DF850488.1 :c495808-494885 Novosphingobium sp. MBES04 DNA, scaffold: MBES4S01 , whole genome shotgun sequence

[0899] ATGTCTGACGCAGATGGCTCCAAGGTCATTTCACGCCACGACGATCCCTCCGCGACGACCGA

[0900] ACGTCCCGGGTACGCGTTCGATCCCACCGATCCCTGGACGATCACGTTCCAGCAAGGCCTCA

[0901] AGGCCGCCGGCCTTGAAGGCAAGGCCGTCTACGAAGTTGGCGTCGGAACCGGCACCAACGT

[0902] GGCCTTCGTGCTTCGCCATTGCGCCGCCAAGGTCTTCTACGGCAGCGACCTCGATCCGCGCC

[0903] TCGTGGAACTGGCCAGGCGCAACGTCGCCAACCTGGCGCCCGAACGCGCTGACAGTTTCCA

[0904] GCCCGTCGAGGGCGCGGTCAGCCTGATCGATACCGATGAGGCCCGCGCCAAGATTGCCAGG

[0905] ACCGATGTCGTGATCGGCTGCCTGCCGCAGGTGGGAGACCCCAATGACGAGCGTTTCGCGG

[0906] CCTTTCGGGCCGAGCACGCGGTCGATCTGCCGCAGGGCGCCGACGATGAGGCCCAGGACCA

[0907] CATCGCCCACTACTACCCCTGGGCGATGTTTGACGAGTATCCCTATAATTCGGTGGGGCTGG

[0908] GGCTGAACGAGGCCCTGTTGCGCCGCATCAAGGAGCAGGCGCCCAAGGCCGACGTGGTGAT

[0909] GAACTTCGGCTGCCGCATCGGGAGTGATCTCATCTTCGAGATGTTCCGTGCCAACGGGTATG

[0910] AGCCCGAAAAGCTGGCGTCGCAGCTTGTCCTGCAACACGCGGGAACCGACATCTCGTTCTTC GTGACGCTGGAAGGTGCCTTGACCGGCACCGATCTGGAGGGCGAGTTCGTCTGCCGCTTCTT TGCCGATCCGCTGGGGCACGAACCTCTCTCGGCCCGCGCGGCACAGGCGCTGCTGGACAAG

[0911] GATCCCAACGTCCCGCTCTACCACGAAGTCGCCGTGATCCGGGGCACCCCCAAGATGGATTG A

[0912] SEQ ID NO: 28 BurB >burB|NC_007650.1 :2553671 -2554189 Burkholderia thailandensis E264 chromosome II, complete sequence

[0913] ATGACGATCACCCTCATCGAACCTGTCCAGCAGCGCACGGGCATCTACCCGTCGTCCGACCT

[0914] GAAGGTCGAGGACGGCTATCCGTCGTCCGACACGTTTCAGATCATCCAGACGCAGGACGGCC

[0915] GCGGTGCGGGCGTGCGCGTGCTGAAAACGTTCGCGCGCGGGCGGCGCATGGCGCGCGTAT

[0916] CCGGACAGATCACCGCGTTCTGCCGGTTGCACACCCTGCAGATCAACGCGCACACGCACCTG

[0917] TACGACCCGCATTTCAGCGGGCTGCTGCTGCATTCGTGCGATCCGAACGTGCGCCTCGACAT

[0918] GGCGGGCTTCGAGCTCTGGTCGCTGCGCGACATCGCGGCGGGCGAGATGCTGACGATGGAC

[0919] TACGCATCGACCGAGGACGTGCTGATGCGCCAGTTCGAATGCCACTGCGGCGCGCCGAACT

[0920] GCCGGCGCTGGATCACGGGCGCGAAGGAGCTGCCGAACGATATCGGGCAGGCGCTCCTCG CCGGATTGCGGGCCGCCGCGCTCGCGTGA

[0921] SEQ ID NO: 29 DSYB >DSYB_euk|KOO32714.1 o- family 2 [Chrysochromulina tobinii]

[0922] MLLVTARRGLFQSRAVLRATPALATAAPALIGARRTLCNSLFELSDDIDVDSVSYGFMASQAIFCAL

[0923] EVGVYEAVAGAGEGGLTADALKTACSINAPRLQTLLTSLTAIKSLKRNAATGAYTLSPNSARYLVQS

[0924] SRQYYGDYLRYQMGRQFYHRMGALPDVMTTGEAPSYASWFSDPATAAMYTQAQHNGSVATAK

[0925] YLVKKKLQLGSVTSMLDVGGGSGAFSYVFVESTPGLKSTVLELPEVCKTGNGIKAKQSPDVQARV

[0926] AFIELDATKPDWPVKEADYELVLMSYISGSVPEAIIGKLYENAFKALKPGGRLLVHDFMVNDSLDGP ALGSLWALQHVTVNAGGLGLTPAEVISRMTAAGFAKCETMEVINGLTKLIVACKE

[0927] SEQ ID NO: 30 >BurE|WP_009894049.1 gamma-aminobutyraldehyde dehydrogenase

[0928] [Burkholderia thailandensis]

[0929] MLINGLCIEGSGEPLSIVDPATGEPLASPAAASAADVERAVAAAEAAFPAWRATTPATRASLLLALA

[0930] DEIERHAHALAQIESRNTGKPLHLWQDEMPAVADCFRFYAGAARTASGPSAGEYVEGHTSMVRR

[0931] DPVGVVAQIAPWNYPLMMAAWKLAPALAAGNTIVFKPSEWTPLSIVALEAALARIFPAGVVNVVLG

[0932] DGANVGRALATHPRVRMISLTGSVEAGKSVLAAAARNLKRTHLELGGKAPVLVFDDADLDAAVAGI

[0933] RYAGFYNAGQDCTAATRIYAQRGVYDALAQRLADAASTLRIGPPDRADAEMGPLVSAAHRARVD

[0934] RFVREAAALPHASVLTGGAPLPGPGCYYAPTVIAGVRHDDAPMRREAFGPVVTLTPFDTESQALR

[0935] WANDSEYGLASSVWTRDAARGMRLAACIEAGVTWVNAHFTYTADMPHGGTKQSGYGSDLSTLG LADYTQPRHVMWRH

[0936] SEQ ID NO: 31 DSYE >DSYE|Frag / 7ar / ops / s_cy / / ndrus_CCMP1102_OEU16654.1_DSYE MAPPTEEENTGDFKDQQFGEKEVADKVVEQYDEKQARVFYKYVMGGGGFDIHYGIFRKATDTVY

[0937] ESSKATNERLLTCLDWTFPVTKDSYVLDLGSGHGGISHEIAARFGCKVMGANISPEQNKMNLLEAK

[0938] KEGVGELVDVTLCNFNDGLPEEWTNKFTHVISCEVLCHAANKPELFKQINRILQPGGAFVFTDIMG

[0939] ADGADEKVLKDFTDRNATTAMARPSEYLTLITDAGLSEPSFLNFSPHLVHYFQSMVNQINENKAAM

[0940] MKEGCSEEYLAKWLESLTSRVDMQREHNVFAWGIFTARTEGNIY

[0941] SEQ ID NO: 32 >burE|NC_007650.1 :2557164-2558576 Burkholderia thailandensis E264 chromosome II, complete sequence

[0942] ATGCTGATTAACGGCCTTTGCATCGAAGGAAGCGGCGAGCCGCTGTCGATCGTCGATCCGGC

[0943] GACGGGCGAGCCGCTCGCATCGCCCGCCGCCGCGAGCGCGGCCGATGTCGAACGCGCGGT

[0944] CGCGGCCGCCGAAGCCGCGTTCCCCGCATGGCGCGCGACGACGCCGGCCACGCGCGCGAG

[0945] CCTGCTGCTCGCGCTCGCCGACGAAATCGAGCGGCACGCGCACGCGCTCGCGCAAATCGAA

[0946] AGCCGCAACACCGGCAAGCCGCTGCATCTCGTCGTGCAGGACGAGATGCCCGCCGTCGCCG

[0947] ACTGTTTCCGCTTCTACGCGGGTGCTGCGCGCACCGCGAGCGGGCCGTCCGCGGGCGAATA

[0948] CGTCGAGGGCCACACGAGCATGGTCCGCCGCGATCCCGTCGGCGTGGTCGCGCAAATCGCG

[0949] CCGTGGAACTACCCGCTGATGATGGCCGCATGGAAACTCGCGCCGGCGCTCGCGGCCGGCA

[0950] ACACGATCGTGTTCAAGCCGTCGGAATGGACGCCGCTGTCGATCGTCGCGCTCGAAGCCGC

[0951] GCTCGCGCGCATCTTTCCGGCGGGCGTCGTCAACGTCGTGCTCGGCGACGGCGCGAACGTC

[0952] GGCCGCGCGCTCGCGACGCATCCGCGCGTGCGGATGATTTCGCTGACGGGCTCGGTCGAG

[0953] GCCGGCAAGTCCGTGCTCGCCGCGGCGGCCAGAAACCTGAAGCGCACGCATCTGGAGCTCG

[0954] GCGGCAAGGCGCCCGTGCTCGTGTTCGACGACGCCGACCTCGACGCGGCCGTCGCCGGCA

[0955] TCCGCTACGCGGGCTTCTACAACGCCGGGCAGGACTGCACGGCGGCCACGCGAATCTACGC

[0956] GCAGCGCGGCGTCTATGACGCGCTCGCGCAGCGGCTCGCCGATGCGGCGAGCACGCTGCG

[0957] CATCGGCCCGCCCGATCGCGCGGACGCCGAGATGGGGCCGCTCGTCAGCGCCGCGCATCG

[0958] CGCGCGCGTCGACCGTTTCGTGCGGGAAGCGGCCGCGCTGCCGCATGCGAGCGTGCTCAC

[0959] GGGCGGCGCGCCGCTTCCCGGCCCCGGCTGCTACTACGCGCCGACCGTCATCGCCGGCGT

[0960] GCGCCACGACGACGCGCCGATGCGCCGCGAAGCGTTCGGCCCGGTCGTCACGCTGACGCC

[0961] GTTCGACACCGAATCGCAAGCGCTCAGGTGGGCGAACGATTCGGAATACGGGCTGGCTTCGT

[0962] CGGTATGGACGCGCGACGCGGCGCGCGGCATGCGGCTCGCCGCGTGCATCGAGGCGGGCG

[0963] TCACGTGGGTGAACGCGCATTTCACCTACACGGCCGACATGCCGCACGGCGGGACCAAGCA

[0964] GTCCGGCTACGGCTCCGATCTGTCGACGCTCGGCCTCGCCGACTACACGCAGCCGCGCCAC

[0965] GTGATGTGGCGGCATTGA

[0966] SEQ ID NO: 33: >DsyB_prok_Roseibium aggregatum LZB033

[0967] MPVARSLETAEEISDIAFGFMGSKALFSALHVDLFSLLSEKTLTPQHVAEESELDLDRATTLLTALTS

[0968] LGLVRREGAGFTNSPAAEAFLVKGRKYDFGDYLRFQIDKQMYPFMTQLNDALTDSLEDDQVASYE

[0969] TWFSDPEEARLYSRSQHAGSLGPGRGLAKLVDLSAAKQLLDVGGGTGAFSISLCKAYPGLRSTVL

[0970] DFPNVAKVGEEFIAEEGLQDRIRYAPGNALKDTWPDSADAVLMSYLFSGVPGTAIPGLVRKAFEVL TPGSDFMVHDFMVDENRDGPKLAALWQLQHTAFNPEARSITSSYVAGLMEAAGFTDIAVEVMIPG MTMLVHGRKPE

[0971] SEQ ID NO: 34 >dsyB_Roseibium aggregatum LZB033

[0972] ATGCCCGTCGCCAGGTCCTTGGAAACGGCTGAGGAAATTTCAGATATTGCTTTCGGATTTATG

[0973] GGCTCCAAGGCTCTTTTTTCTGCACTTCATGTAGATTTGTTTTCACTGCTTTCCGAAAAGACCT

[0974] TGACGCCACAGCATGTTGCTGAAGAAAGCGAGCTTGATCTTGATCGCGCCACAACACTGTTGA

[0975] CCGCGCTTACGAGCCTCGGGCTGGTGCGCCGGGAGGGAGCAGGCTTCACCAATTCGCCCGC

[0976] AGCGGAGGCGTTTCTGGTGAAAGGACGGAAGTATGATTTCGGCGACTATCTGCGGTTTCAGA

[0977] TCGACAAGCAGATGTACCCCTTCATGACGCAGCTCAATGACGCCCTGACGGACAGTCTGGAG

[0978] GACGACCAGGTTGCTTCCTATGAGACCTGGTTCAGCGACCCGGAAGAAGCACGGCTTTATTC

[0979] CCGGTCCCAGCATGCAGGCTCGCTTGGGCCCGGGCGGGGACTTGCCAAGCTTGTGGATCTG

[0980] TCCGCTGCCAAGCAGTTGCTGGACGTTGGCGGTGGCACCGGGGCCTTCTCGATTTCGCTTTG

[0981] CAAGGCGTATCCGGGACTGCGATCAACCGTTCTGGATTTTCCGAATGTCGCCAAGGTAGGTG

[0982] AAGAGTTCATTGCCGAAGAAGGCCTGCAGGACCGCATCCGCTATGCGCCGGGCAATGCCCTG

[0983] AAGGACACATGGCCGGACAGCGCGGACGCAGTGCTGATGTCCTATCTCTTTTCCGGCGTCCC

[0984] GGGCACGGCCATTCCCGGGCTTGTGCGCAAGGCCTTCGAGGTTCTGACACCGGGCAGTGAC

[0985] TTCATGGTGCATGACTTCATGGTCGACGAAAACCGCGACGGGCCGAAGCTGGCGGCACTCTG

[0986] GCAGCTTCAGCACACCGCCTTCAACCCGGAAGCCCGTTCGATCACGAGTTCCTACGTTGCCG

[0987] GCTTGATGGAAGCCGCCGGCTTCACAGATATCGCAGTTGAAGTGATGATCCCGGGCATGACG

[0988] ATGCTGGTGCACGGCCGCAAGCCGGAATAG

[0989] SEQ ID NO: 35 >DsyB_prok_Amorphus_coralli_DSM_19760

[0990] MTLLTTAEEISDIAFGFMGSKALFAALHFGVFTHLAERPMTAEELGQAAGLPAERARTLLTAVASLG

[0991] LVSVEDGRFANAPAAEAFLVKGAKYDFGDYLRLQVDRQMYGLLDQIEPALANRLPEDATGSYAEW

[0992] FSDPEQARIYSESQHAGSLGPARGLAKSVDLAGARTLLDVGGGTGAYAITLCKANPDLAATVVDFP

[0993] NVAALGREYVAEAGLADRVSYVDGNALETDWPAGQDSVLMSYLFSGVPGEEHDRLVRRAYDTLT

[0994] PGGLYMVHDFVVDADRTGPKLAALWQLQHTAFTPTARSLDEATLADMMTGAGFEGVEVREMIPG LTMLATGRKPG

[0995] SEQ ID NO: 36 >dsyB_Amorphus_coralli_DSM_19760

[0996] ATGACCCTCCTGACCACGGCCGAGGAGATCTCCGACATCGCCTTCGGCTTCATGGGCTCGAA

[0997] GGCCTTGTTCGCCGCGCTTCATTTCGGCGTGTTCACCCATCTCGCCGAGAGGCCGATGACGG

[0998] CGGAGGAGCTTGGACAGGCCGCCGGCCTGCCGGCGGAGAGGGCACGGACGCTGCTGACGG

[0999] CGGTCGCCTCCCTCGGCCTGGTGTCGGTCGAGGACGGCCGCTTCGCCAACGCGCCGGCGG

[1000] CGGAGGCCTTCCTGGTGAAGGGCGCGAAGTACGATTTCGGCGACTACCTGCGTCTGCAGGT

[1001] GGATCGGCAGATGTACGGACTGCTCGACCAGATCGAACCGGCGCTGGCCAATCGTCTGCCC

[1002] GAGGATGCGACCGGCAGCTACGCCGAGTGGTTCTCCGATCCCGAGCAGGCCCGGATCTACT CGGAAAGCCAGCACGCTGGTTCGCTCGGTCCGGCGCGAGGCCTTGCCAAGAGCGTCGACCT

[1003] GGCCGGCGCGAGGACGCTCCTCGACGTGGGGGGTGGCACCGGCGCCTACGCCATCACGCT

[1004] CTGCAAGGCCAATCCGGACCTAGCCGCGACGGTGGTCGACTTCCCGAACGTCGCCGCGCTC

[1005] GGCCGAGAGTATGTCGCCGAGGCGGGGCTCGCTGACCGGGTTTCCTATGTCGACGGCAACG

[1006] CGCTGGAGACGGACTGGCCGGCCGGGCAGGACTCGGTCCTGATGTCCTACCTCTTCTCCGG

[1007] CGTGCCGGGTGAAGAGCACGACCGCCTGGTGCGTCGCGCTTATGACACCCTGACGCCCGGC

[1008] GGCCTGTATATGGTCCACGATTTCGTGGTGGACGCGGATCGGACCGGCCCCAAGCTCGCCG

[1009] CTCTCTGGCAGCTCCAGCACACGGCCTTCACGCCGACGGCGCGCAGTCTCGACGAGGCAAC

[1010] GCTCGCCGACATGATGACCGGCGCGGGCTTCGAGGGTGTCGAGGTGCGGGAAATGATCCCC

[1011] GGTCTGACCATGCTGGCTACGGGCCGCAAGCCGGGTTGA

[1012] SEQ ID NO: 37 >DsyB_prok_Roseibium_aggregatum_IAM_12614

[1013] MPVARSLETAEEISDIAFGFMGSKALFSALHVDLFSLLSEKTLSPDEVSRKSELDLDRATTLLTALAS

[1014] LGLVRREGTGFTNSPAAEAFLVKGRKYDFGDYLRFQIDKQMYPFMTQLNDALTDSLEDGQVASYE

[1015] DWFSDPEEARLYSRSQHAGSLGPGRGLAKLVDLSAAKQLLDVGGGTGAFSISLCKAYPGLRSTVL

[1016] DFPNVAKVGEEFIAEEGLQDRIQYAPGNALKDPWPDSADAVLMSYLFSGVPGTAIPGLVRKAFEVL

[1017] TPGGDLMVHDFMVDENRDGPKLAALWQLQHTAFNPEARSITSSYVAGLMEAAGFIDIAVEVMIPG MTMLVHGRKPD

[1018] SEQ ID NO: 38 >dsyB_Roseibium_aggregatum_IAM_12614

[1019] ATGCCTGTCGCCAGGTCCTTGGAAACGGCTGAGGAAATTTCAGATATTGCTTTTGGGTTCATG

[1020] GGCTCGAAAGCCCTTTTTTCTGCACTTCATGTGGATTTGTTTTCACTTCTTTCCGAAAAGACCT

[1021] TGTCTCCAGACGAAGTTTCTCGGAAAAGCGAACTTGATCTTGATCGCGCGACAACCTTGCTGA

[1022] CTGCTCTCGCCAGCCTTGGGCTGGTGCGCCGGGAGGGAACCGGCTTTACCAATTCTCCTGCG

[1023] GCGGAAGCATTTCTGGTGAAGGGCCGGAAATATGATTTCGGCGACTATCTGCGGTTCCAGAT

[1024] CGACAAGCAGATGTATCCCTTCATGACGCAGCTCAACGATGCGCTGACCGACAGCCTGGAGG

[1025] ATGGACAGGTAGCCTCCTATGAGGACTGGTTCAGCGACCCGGAGGAAGCGCGGCTTTATTCA

[1026] AGGTCCCAGCACGCTGGCTCGCTCGGGCCCGGGCGGGGGCTCGCCAAGCTTGTGGATCTGT

[1027] CCGCTGCCAAGCAGTTGCTGGACGTCGGCGGCGGCACCGGGGCGTTCTCGATTTCGCTCTG

[1028] CAAGGCTTATCCCGGATTGCGGTCAACCGTTCTGGACTTCCCCAATGTTGCCAAGGTGGGCG

[1029] AGGAGTTCATTGCCGAGGAAGGCTTGCAGGATCGCATCCAGTATGCGCCGGGCAATGCCCTG

[1030] AAGGACCCTTGGCCGGATAGCGCGGACGCGGTGCTGATGTCTTACCTCTTTTCCGGTGTGCC

[1031] GGGCACGGCCATTCCGGGGCTTGTGCGCAAGGCGTTCGAGGTTCTGACACCGGGCGGCGAT

[1032] TTGATGGTCCATGACTTCATGGTCGACGAAAATCGCGACGGGCCGAAGCTGGCTGCACTGTG

[1033] GCAATTGCAGCATACCGCCTTCAATCCGGAAGCCCGCTCGATCACGAGTTCCTATGTTGCCG

[1034] GTTTGATGGAAGCTGCCGGCTTCATCGATATCGCGGTTGAGGTGATGATCCCGGGAATGACG

[1035] ATGCTGGTGCATGGCCGCAAGCCGGACTGA SEQ ID NO: 39>DsyB_prok_Pseudooceanicola_batsensis_HTCC2597

[1036] MHPATEADEISAIAFGFMGSKALFVALDLGVFTKLAGGSATAEEMAQATGIHRDRAETLLTALTGLG LLTVKAGRFANSPAADSFLVKGAKYDFGDYLRLQVGRQMYGLLDQLDAAVQGEMTEGATASYEQ WFSDPDQARLYSESQHAGSLGPARQLAKKVDLSGARRLLDVGGGTGAFAITLCRAFPELTATVVE

[1037] FPNVATLGRKYVEEAGLSDRITYVEGNALSTDWPEGQDTVLMSYLFSGVPGDAHTDLIADARAAL

[1038] APGGQVVIHDFMVEADRSGPELAALWQLQHTAFTPEARSVDTGTLAEELTQGGFEKVDIVEMIPQ MTKVAVGRRAA

[1039] SEQ ID NO: 40 >dsyB_Pseudooceanicola_batsensis_HTCC2597

[1040] ATGCATCCAGCGACCGAAGCCGACGAGATTTCCGCCATTGCATTCGGGTTCATGGGGTCCAA

[1041] GGCGCTCTTCGTCGCGCTCGACCTCGGCGTGTTCACGAAGCTGGCCGGAGGGTCGGCAACG

[1042] GCCGAAGAGATGGCACAGGCCACGGGTATCCACAGGGATCGGGCCGAAACCCTCCTGACGG

[1043] CGCTGACCGGCCTGGGGCTGCTGACGGTGAAGGCGGGCCGTTTCGCCAATTCGCCGGCCGC

[1044] CGATTCGTTTCTGGTGAAGGGGGCCAAGTACGATTTCGGCGATTACCTGCGGCTGCAGGTCG

[1045] GGCGCCAGATGTACGGGCTTCTGGACCAGCTGGACGCGGCGGTGCAGGGCGAGATGACCG

[1046] AGGGTGCCACGGCGAGTTACGAACAGTGGTTCTCGGATCCCGACCAGGCCCGGCTCTATTCC

[1047] GAGAGCCAGCACGCCGGATCGCTGGGCCCTGCGCGGCAACTGGCGAAAAAGGTGGACCTGT

[1048] CGGGCGCGCGGCGGCTGCTGGACGTCGGCGGCGGGACCGGTGCCTTCGCCATCACCTTGT

[1049] GCAGGGCCTTTCCCGAGCTGACGGCGACGGTGGTCGAATTTCCCAATGTCGCCACGCTGGG GCGGAAATACGTCGAGGAGGCGGGGCTGTCGGACCGGATCACCTATGTCGAGGGCAACGCG CTGTCGACGGACTGGCCGGAGGGGCAGGACACGGTGCTGATGTCCTACCTGTTCTCGGGGG

[1050] TGCCGGGCGACGCGCACACCGACCTGATCGCGGATGCCCGGGCGGCGCTGGCGCCCGGGG

[1051] GGCAGGTGGTGATCCACGACTTCATGGTCGAGGCCGACCGAAGCGGGCCGGAACTCGCCGC

[1052] GCTCTGGCAGTTGCAGCACACGGCCTTCACGCCGGAAGCGCGGTCGGTGGATACGGGGACG

[1053] CTGGCAGAGGAACTGACGCAGGGCGGGTTCGAGAAGGTCGACATCGTCGAGATGATCCCGC

[1054] AGATGACCAAGGTCGCCGTGGGGCGCCGGGCGGCGTGA

[1055] SEQ ID NO: 41 >DsyB_prok_Salipiger_bermudensis _HTCC2601

[1056] MGPVTDADEISRIAFGFMGSQALFTALDHGLFTVLAEGALDAEALADRTGLHRDRAETLLTALAGL GLVTVSDGRFANSPAAEAFLVKGAKYDFGDYLRLQVGKQMYGLMGQLGDAVSGALGEGATASYE QWFSDPEQARLYSESQHAGSLGPARQLAKRIDLSGARQLLDVGGGTGAFAITLCKAFPELSATIVD

[1057] FPNVAALGRRHVAEAGLSDRIAYVEGNALETDWPGGQDVVLMSYLFSGVPGSAHEGLLRAAHDR LMPGGRLLIHDFVVHADRSGPPLAALWQLQHTAFTPEARSVDAEGLARDLWAAGFAEVTVSEMIP QMTMLAEARRPE

[1058] SEQ ID NO: 42 >dsyB_Salipiger_bermudensis_HTCC2601

[1059] ATGGGGCCGGTCACGGACGCCGACGAGATTTCGCGAATTGCCTTCGGGTTCATGGGGTCACA GGCGCTGTTCACTGCGCTGGATCATGGCCTCTTTACCGTGCTCGCCGAGGGCGCGCTTGATG CAGAGGCGCTGGCAGACCGCACCGGCCTGCATCGCGATCGCGCCGAGACGCTGCTGACCG

[1060] CGCTGGCAGGGCTGGGGCTGGTGACGGTCTCGGACGGGCGCTTTGCGAATTCACCGGCCGC

[1061] CGAGGCCTTTCTGGTGAAGGGCGCGAAATACGATTTCGGCGATTACCTGCGGCTTCAGGTCG

[1062] GCAAGCAGATGTACGGGCTCATGGGCCAGCTCGGTGACGCAGTCTCGGGGGCGCTGGGTGA

[1063] GGGGGCCACGGCCTCCTACGAGCAATGGTTCTCCGACCCAGAGCAGGCGCGGCTCTATTCC

[1064] GAGAGCCAGCATGCGGGCTCGCTCGGCCCGGCGCGCCAGCTCGCCAAGCGCATCGACCTGT

[1065] CGGGGGCGCGCCAGCTGCTCGACGTGGGCGGCGGAACTGGCGCCTTCGCGATCACGCTGT

[1066] GCAAGGCCTTCCCGGAGCTCTCGGCCACCATCGTCGATTTCCCCAATGTCGCGGCGCTCGGC

[1067] CGCCGCCATGTCGCCGAGGCAGGTTTGTCGGACCGGATCGCGTATGTCGAGGGGAACGCGC

[1068] TCGAAACCGACTGGCCGGGCGGGCAGGACGTGGTGCTGATGTCCTACCTGTTCTCCGGCGT

[1069] GCCTGGATCGGCGCATGAGGGGCTGCTCCGCGCCGCCCATGATCGGCTGATGCCGGGCGG

[1070] ACGGCTGTTGATCCACGATTTCGTGGTCCATGCCGACCGCAGCGGCCCGCCGCTGGCGGCG

[1071] CTCTGGCAGCTCCAGCACACCGCCTTCACGCCCGAGGCGCGCTCGGTCGATGCCGAAGGGC

[1072] TGGCGAGGGATCTGTGGGCGGCGGGCTTTGCCGAGGTGACGGTCTCCGAGATGATCCCGCA

[1073] GATGACGATGCTGGCCGAAGCGCGCCGACCGGAGTGA

[1074] SEQ ID NO: 43 >DsyB_prok_Sediminimonas_qiaohouensis_DSM_21189

[1075] MTLATKADEISEIAFGFMGSKALFAALQLKVFTHLAEGPLSAEELAQKANVHPDRAQTLLTALASLG

[1076] LIEVQDGNRFGNAPASQAFLVNGAKYDFGDYLRLQVGRQMYSLLDQIENALQGTMDADDTASYA

[1077] EWFADPDEARLYSESQHSGSVGPARQLSGALDLSGARRMLDVGGGTGAFAITLCKDNPDLTASIV

[1078] DFPNVAELGRGYVEKAGLSDRIAYIPGNALEAEWPADQDVILMSYLLSGVPGETHVDLIRRAYDHL

[1079] VPGGRLLIHDFVVEKERTGPKLAALWQLQHTAFTPEARSLDAGWLEVALDEIGFIDAQVAPLIPKMT MLAQGTKPAA

[1080] SEQ ID NO: 44 >dsyB_Sediminimonas_qiaohouensis_DSM_21189

[1081] ATGACACTTGCCACCAAGGCAGATGAAATTTCCGAGATCGCGTTCGGCTTCATGGGGTCCAA

[1082] GGCGCTGTTCGCGGCGCTGCAACTCAAGGTGTTTACCCACCTTGCAGAAGGGCCACTGAGCG

[1083] CCGAGGAACTGGCGCAAAAGGCCAATGTGCACCCCGACCGCGCGCAAACCCTGCTGACCGC

[1084] GCTGGCCTCCCTGGGCCTGATCGAGGTCCAGGATGGCAACCGCTTCGGCAATGCGCCCGCG

[1085] TCGCAGGCGTTCCTGGTCAACGGGGCAAAGTACGATTTCGGCGATTATCTGCGGTTGCAGGT

[1086] GGGGCGCCAGATGTACAGCCTGCTGGACCAGATCGAGAACGCCCTGCAAGGCACGATGGAT

[1087] GCCGATGATACCGCCAGCTATGCCGAGTGGTTCGCCGATCCCGACGAGGCGCGGCTTTACTC

[1088] CGAAAGCCAGCATTCCGGCTCGGTCGGGCCGGCACGGCAACTGTCGGGCGCGCTTGACCTG

[1089] TCGGGCGCGCGGCGCATGCTGGACGTTGGCGGCGGCACCGGTGCGTTCGCCATCACCCTGT

[1090] GCAAGGACAATCCGGATTTGACTGCGTCGATCGTCGATTTCCCCAACGTGGCCGAGCTGGGC

[1091] CGCGGCTATGTGGAGAAGGCCGGTTTGTCGGACCGGATCGCGTATATCCCCGGCAATGCGC

[1092] TCGAGGCCGAGTGGCCCGCCGATCAGGACGTGATCCTGATGTCCTACCTGCTTTCGGGTGTG

[1093] CCCGGCGAGACGCATGTCGATCTGATCCGCCGCGCCTATGATCACCTGGTGCCGGGTGGGC GGCTGCTGATCCACGACTTCGTCGTCGAGAAAGAGCGCACCGGCCCCAAGCTGGCCGCGCT

[1094] GTGGCAGTTGCAGCATACCGCGTTCACGCCCGAGGCGCGCTCGCTCGATGCCGGCTGGCTC

[1095] GAGGTGGCGCTGGACGAGATCGGGTTTATCGATGCGCAGGTGGCACCGCTCATCCCCAAGA

[1096] TGACGATGCTGGCGCAGGGCACCAAGCCCGCCGCATAA

[1097] SEQ ID NO: 45 >DsyB_prok_Sagittula_stellata_E-37

[1098] MAVLTEAEDISDIAFGYMGSKALFAALEFGVFTALSQGNIGLTGIAGATGLPKERCRTLLSALVGLG

[1099] LVTHDDAGFANSPAAESFLVKGARHDFGDYLRLQVGRQMYPLMDQIEKALTGDLEDDHTGSYAQ

[1100] WFADPEEARLYSESQHAGSLGPARGLAKRVDFSGIGSLLDVGGGTGAFAITLARRNPNLRITVLDF

[1101] PNVAKLGEAYVADAGLSSQIGYCHGNALESGWPGGQDAVLMSYLFSGVPDHSHAGLLRKAHDAL

[1102] NPGGQVLIHDFIVDADLSGPKNTALWQLQHTAFTPEARSLDDDWLIGALEAAGFSDADVGPLIPGM TKLATARKA

[1103] SEQ ID NO: 46 >dsyB_Sagittula_stellata_E-37

[1104] ATGGCGGTCCTGACCGAGGCAGAGGACATTTCGGATATCGCGTTTGGCTACATGGGCTCCAA

[1105] GGCGTTGTTTGCGGCGCTCGAATTCGGTGTGTTTACGGCCTTGTCACAGGGCAACATCGGAT

[1106] TGACTGGAATTGCCGGGGCCACGGGCCTGCCCAAGGAGCGTTGCCGCACCCTGCTGTCCGC

[1107] GCTGGTGGGTTTGGGCCTTGTCACACATGACGACGCTGGGTTCGCCAATTCCCCGGCGGCG

[1108] GAGTCTTTCCTTGTGAAAGGGGCGCGTCACGACTTCGGCGACTACCTGCGCCTGCAGGTCGG

[1109] CCGCCAGATGTATCCGCTGATGGACCAGATCGAGAAGGCGTTGACCGGGGACCTGGAAGAC

[1110] GATCACACTGGATCCTACGCACAGTGGTTTGCCGACCCGGAAGAGGCCCGGCTGTATTCCGA

[1111] GAGCCAGCACGCCGGATCGCTCGGTCCGGCGCGGGGGCTGGCCAAGCGGGTCGACTTTTC

[1112] GGGCATCGGCAGCCTGCTGGATGTGGGCGGCGGGACCGGCGCCTTTGCCATCACGCTGGC

[1113] GCGCAGGAACCCGAACCTGCGCATCACGGTGCTGGACTTCCCCAATGTCGCCAAGCTGGGC

[1114] GAAGCCTACGTGGCTGATGCGGGACTGTCGTCGCAAATCGGCTATTGCCATGGGAACGCGCT

[1115] GGAGTCTGGATGGCCCGGCGGGCAGGATGCCGTCCTGATGAGCTACCTGTTTTCGGGTGTG

[1116] CCGGATCACAGCCACGCGGGCCTGCTGCGCAAGGCACATGACGCGCTCAATCCCGGCGGGC

[1117] AGGTGCTGATCCACGACTTCATCGTCGACGCGGACCTGTCCGGCCCGAAGAACACAGCGCTC

[1118] TGGCAGCTTCAGCACACGGCCTTCACCCCCGAGGCCCGATCGCTGGACGACGACTGGCTGA

[1119] TCGGCGCCCTTGAGGCCGCAGGGTTCAGCGACGCCGATGTCGGGCCGCTGATCCCCGGGAT

[1120] GACCAAGCTGGCCACCGCCCGAAAGGCCTGA

[1121] SEQ ID NO: 47 >DsyB_prok_Thalassobaculum_salexigens

[1122] MKPIESAEDISELAFGFMASKALFAALHVDVFGALSDGPKSITDLAAATKVPAQRMQTLVTALVSVG

[1123] LLTRNDGKIANAPASDAYLVRDNTNYFGDYLRFQIDRQMYPFMENLDKVLLGDTDDIEYPDYASW

[1124] MADRHHAELFSRSQHSGSLGPGAVLAKRLLKEGAVSEDVGSMLDVGGGSGAFSIMFCKRFPKLH

[1125] ATVLDFPNVIEVGKTFVAEEEMSDRIDFVAGDGTNANWPNDQDIVLMSYLFSGVPEEAIDKLCSDA FRVLKPGGLIAIHDFMVTDDRKGPALAALWQLQHMVYTPDGVGMTPGFVEKHLKKAGFEIEIDDDL

[1126] IPGMTRVMTARKPG

[1127] SEQ ID NO: 48 >dsyB_Thalassobaculum_salexigens

[1128] GTGAAACCGATCGAATCCGCCGAGGACATTTCCGAGCTCGCCTTCGGGTTCATGGCGTCGAA

[1129] GGCTCTGTTCGCGGCCCTGCATGTGGACGTGTTCGGGGCACTGTCCGACGGCCCGAAATCG

[1130] ATCACGGATCTGGCGGCGGCGACGAAGGTGCCCGCGCAGCGCATGCAGACGCTGGTGACG

[1131] GCCCTGGTCTCGGTCGGCCTGCTCACCCGCAACGACGGCAAGATCGCCAACGCGCCGGCCA

[1132] GCGACGCCTATCTGGTGCGCGACAACACCAACTATTTCGGCGATTACCTGCGCTTCCAGATC

[1133] GATCGCCAGATGTATCCGTTCATGGAGAACCTGGACAAGGTTCTGCTCGGCGACACCGACGA

[1134] CATCGAGTATCCGGACTACGCGTCCTGGATGGCCGACCGGCACCATGCCGAGCTGTTCAGCC

[1135] GCTCGCAGCATTCCGGCTCGCTCGGCCCGGGCGCGGTGCTGGCCAAGCGCCTGCTCAAGGA

[1136] AGGCGCGGTCTCCGAGGATGTCGGCTCGATGCTTGATGTCGGCGGCGGCTCCGGCGCGTTC

[1137] TCCATCATGTTCTGCAAGCGCTTCCCGAAACTGCACGCCACGGTGCTCGACTTCCCCAACGT

[1138] GATCGAGGTCGGCAAGACCTTCGTCGCCGAGGAGGAGATGTCGGACCGGATCGACTTCGTC

[1139] GCCGGCGACGGCACCAACGCGAACTGGCCGAACGACCAGGACATCGTGCTGATGAGCTATC

[1140] TGTTCTCCGGCGTGCCGGAGGAGGCGATCGACAAGCTGTGCTCCGATGCGTTCCGGGTGCT

[1141] GAAGCCGGGCGGGCTGATCGCGATCCACGACTTCATGGTGACCGACGACCGCAAGGGCCCG

[1142] GCCCTGGCGGCGCTGTGGCAGCTCCAGCACATGGTCTACACGCCGGACGGTGTCGGCATGA

[1143] CCCCGGGCTTCGTGGAGAAGCACCTGAAGAAGGCCGGGTTCGAGATCGAGATCGACGACGA

[1144] CCTCATCCCCGGCATGACCCGGGTGATGACCGCCCGCAAGCCGGGCTGA

[1145] SEQ ID NO: 49 >DSYB_euk_Prymnesium_parvum_CCAP_946 / 6

[1146] MLRLAPRLPTRALVRHALRAHALPLARPALPSGTRLFASAPADDIDVDNVAYGFMASQALFTGLEM

[1147] GLFDAIAAGPEAGLNIDELKAAANCSAPRLQTLVTSLVAIKSLKRTPDGRYTLSPNTARFLVQSSKQ

[1148] YYGDYLKYQMGRQFYHRMGALPDVMTSGEAPSYASWFSDPETAATYTQAQHNGSVATAKYLIKK

[1149] KLQLGDAATMLDVGGGSGAFSYVFTEATPGLSSTVLELPEVCRTGEAIKAKQPVSVQERVKLVEL

[1150] DATSPDWPVNDAAYDWLMSYISGSVPESIIGALYANAYKALKPGGRLLVHDFMVDNSLDGPPLGA

[1151] LWALQHVTVNADGLGLCPQGVIERMGSAGFDPSACETMEMITGLTKLIVAYKP

[1152] SEQ ID NO: 50 >DSYB_euk_Chrysochromulina_tobinii_CCMP291

[1153] MLLVTARRGLFQSRAVLRATPALATAAPALIGARRTLCNSLFELSDDIDVDSVSYGFMASQAIFCAL

[1154] EVGVYEAVAGAGEGGLTADALKTACSINAPRLQTLLTSLTAIKSLKRNAATGAYTLSPNSARYLVQS

[1155] SRQYYGDYLRYQMGRQFYHRMGALPDVMTTGEAPSYASWFSDPATAAMYTQAQHNGSVATAK

[1156] YLVKKKLQLGSVTSMLDVGGGSGAFSYVFVESTPGLKSTVLELPEVCKTGNGIKAKQSPDVQARV

[1157] AFIELDATKPDWPVKEADYELVLMSYISGSVPEAIIGKLYENAFKALKPGGRLLVHDFMVNDSLDGP

[1158] ALGSLWALQHVTVNAGGLGLTPAEVISRMTAAGFAKCETMEVINGLTKLIVACKE SEQ ID NO: 51 >CDS_DSYB_euk_Chrysochromulina_tobinii_CCMP291

[1159] ATGCTGCTCGTCACTGCTCGGCGAGGCCTCTTCCAGTCGCGTGCAGTCTTGCGGGCGACGC

[1160] CCGCCCTCGCGACGGCCGCGCCGGCGCTGATTGGCGCACGTCGCACTCTCTGCAACTCGCT

[1161] GTTCGAGTTGTCCGACGACATTGACGTGGACAGCGTGTCATATGGCTTCATGGCATCGCAGG

[1162] CGATCTTTTGCGCGCTCGAGGTGGGCGTATACGAGGCAGTTGCGGGCGCGGGCGAGGGCG

[1163] GCCTTACGGCCGATGCCCTCAAGACTGCGTGCAGCATAAACGCGCCGCGGTTGCAGACGCT

[1164] GCTCACGTCGCTCACTGCGATCAAGTCGCTCAAGCGCAACGCCGCCACGGGCGCTTACACG

[1165] CTCTCGCCCAACAGCGCACGCTACCTTGTCCAGTCGTCGCGCCAATACTACGGCGACTACCT

[1166] GCGCTACCAGATGGGTCGCCAGTTCTACCACCGCATGGGCGCGCTGCCAGATGTGATGACG

[1167] ACCGGCGAGGCACCGTCATACGCGTCGTGGTTCTCGGACCCGGCCACCGCGGCGATGTACA

[1168] CGCAGGCGCAGCATAACGGCTCGGTCGCCACGGCCAAGTACCTTGTCAAGAAAAAGCTTCAG

[1169] CTCGGCTCGGTCACGTCAATGCTCGACGTGGGCGGCGGCTCGGGCGCCTTTTCGTACGTCTT

[1170] CGTCGAGAGCACTCCGGGACTCAAGTCGACAGTCCTCGAGCTGCCTGAGGTGTGCAAGACG

[1171] GGCAACGGCATCAAGGCGAAGCAGTCGCCCGATGTGCAGGCACGCGTCGCATTCATCGAGC

[1172] TCGACGCGACGAAGCCCGACTGGCCGGTGAAGGAAGCGGACTATGAGCTTGTGCTCATGAG

[1173] CTATATATCGGGTTCGGTGCCCGAGGCCATCATCGGCAAGCTGTACGAGAACGCGTTCAAGG

[1174] CTCTCAAGCCGGGGGGTCGGCTGCTCGTGCACGATTTTATGGTGAACGACTCGCTCGACGGC

[1175] CCAGCGCTTGGTTCGCTCTGGGCGCTGCAGCACGTGACGGTCAACGCCGGCGGACTCGGTC

[1176] TGACCCCCGCGGAGGTCATCAGCCGCATGACCGCGGCGGGCTTTGCCAAGTGCGAGACGAT GGAGGTGATCAATGGCCTGACTAAGCTGATCGTCGCGTGCAAGGAGTGA

[1177] SEQ ID NO: 52 >DSYB_euk_Lingulodinium_polyedrum_CCMP1936

[1178] MAFAPRTSPLLARAVTRAWPRAQRSAAAALRSWARPALAAKPPLASRAFARTSWDEGEDVDLDS VAYGFMASQALFTGLELGIFDHIAAAGAGGLSAAGIGKACGIEAPRVQTLLTSLVAVKCLKRDASAM YTLSPNTAQYMVTSSRHFYGDYLRYQIGRQFYHRMGALPEVMTSGKAPSYASWFSDPEVARTYT

[1179] QAQHNGSVATAKYLIKKKLQLGGISAMLDVGGGSGAFSYVFTQATPGLHSKVLELPEVCRTGEGIR AKQPEDVRSRVSFVELDASSPTWPVDDSAFDVVLMSYISGSVPEPIIGSLYANAMKALRPGGRLLV HDFMVNDSLDGPALGALWGLQHVTVNADGLGLCPKEIISRMGTAGFDTSKCETMEMIHGMTKLIV

[1180] GYKN

[1181] SEQ ID NO: 53 >DSYB_euk_Alexandrium_tamarense_ATSP1 B_1

[1182] MPPSRAWRRLAVSAQQILPSIVQGTSSEDVDLDALAYGFMASQALFSALELGIFDHLAACPATAPE LGSACGIPTQRLQTLLTALVASRCLRLDAESLKYTNSPNVARFMVSNSKSYYGDYLKCQIGWLFYH RMGRITEVMKGGEALDYQTWFSDPHVADTYTSAQHNGSLATARALMRKVDLSRVTCLLDAGGGS

[1183] GAFSIAAARAVPGLEATVLELPEVCKTGSRIVEQAGLSGRIRYVELDATSPDWPVHGGSFQAVLMS YLSGSVPAHAIVGLYSNAYKALGPGGRLVVHDFMVDNSLDGPQLGALWALQHVTVNPDGLGLCP GHWSRMRTAGFARTETLDLIGGMTKVVVGYKD SEQ ID NO: 54 >DSYB_euk_Acropora_cervicornis

[1184] MAAAFAAARGLTRAISRCASKRVPEQALFSRLSAPAPPALAALPGVRAFSRTSWESGDDVDLDSV

[1185] AYGFMASQALFSALELGIFDKVAAAGEKGCAAKDVQQACGVEGPRLTTLLTALTAVKCLRRSDEG

[1186] LYTLSPNTAQYMVSSSRHYYGDYLQYQIGRQFYHRMGALPEVMTTGKAPSYASWFSDPEVAKTY

[1187] TQAQHNGSVATAKYLVRKKLDLGGISSMLDVGGGSGAFSYVFTEATPGLKSTVLELPEVCRTGEGI

[1188] KAQQPQDIQDRVSFVELDATSPDWPVSDSNYDIVLMSYISGSVPESVILPLYKNAFKALRPGGRLL

[1189] VHDFMVNDSLDGPALGALWGLQHVTVNAQGLGLCPAEVIRRMAQAGFEENKCQTHEMIHGMTKL IVAHKA

[1190] SEQ ID NO: 55 >DSYB_euk_Fragilariopsis_cylindrus_CCMP1102

[1191] MTLSTPTTVGSGEDEVSKKNDDQPAIDDIAYGFMGSKALSVALKIGVFDAIDVVSKGEDNAVEGGA

[1192] TLNQILTHCKVPKERLRTLLSACVALKLINRRVLRGEDVFSLPEASAEQLVKSSKRYWGDYIVGQVD

[1193] AQFYVRMKDLDTTILTGDTSSDGYEAWFDKDPDAAKRYTQAQHNGSLATGYGLLKRLPELSQGE

[1194] KYPNLRMLDIGGGSGAFSIATARKVKDADCVILDLPNVIKVAEEIISKEDEQVRNRLSTVALSAADPG

[1195] NWMIEDESFDVVLMSYVSGSISSDALSGLYQNAFRALRPGGMVVIHDFFVDNNGEGPKKAALWAL THVTVNPEGMGLRPGRIVKLLTENGLVGPKVEDMIPGTTQLIVATKPKLA

[1196] SEQ ID NO: 56 >DSYB_euk_Symbiodinium_sp._KB8

[1197] MSRPIRRLKVVTGHTTALRTSACGSEDVDLDSLAYGFMASQALFAALELGVFDYLAKGPQSVSDL

[1198] AEGCGVPCNRLQTLLTALVAAKCLRRDESQLYSNSPNVQKFMVSTSKAYYGDYFKHQVGGLFYA

[1199] RMGQLAKVLRGEEVLDYSQWFSDPTVASLYTSAQHNGSLATAKSLFRKVSLATTGRMLDVGGGS

[1200] GAFSLQAAQMNPELRATVLDLPEVCRVGRSLMEQAASPEKSRVGFKELDATSPAWPVKAESQDL

[1201] ALMSYLCGSVPEDVIAQLFQNAFRVLRSGGRLVVHDFMVDDTRDGPTLGAYWALQHVTVNPTGL

[1202] GLAPCDISSRMKAAGFDHVEVFDMIAGMTKVVLATKP

[1203] SEQ ID NO: 57 >CDS_DSYB_euk_Symbiodinium_sp._KB8

[1204] ATGTCGCGCCCGATCCGCCGATTGAAGGTTGTGACGGGCCATACTACCGCTCTACGGACCTC

[1205] AGCTTGTGGAAGCGAGGATGTCGACCTCGACAGCCTGGCCTATGGGTTCATGGCTTCGCAGG

[1206] CTCTCTTTGCCGCGCTGGAACTGGGCGTCTTCGATTACCTGGCGAAGGGCCCGCAAAGCGTC

[1207] TCCGACTTGGCAGAAGGCTGCGGGGTGCCCTGCAATCGCCTGCAGACTTTGCTGACAGCGCT

[1208] CGTGGCAGCAAAGTGCTTGCGCCGAGACGAATCACAGCTCTACAGTAATTCGCCGAATGTTC

[1209] AAAAGTTTATGGTGTCGACTTCAAAGGCGTACTATGGTGACTATTTCAAACATCAGGTGGGTG

[1210] GCTTGTTTTATGCCCGGATGGGGCAGCTGGCAAAAGTGCTGAGGGGAGAGGAGGTACTCGA

[1211] CTACTCCCAGTGGTTCTCGGATCCAACAGTGGCCTCCCTC...

Claims

Claims1 . A method of altering a plant stress response comprising contacting a plant or part thereof with a formulation comprising dimethylsulfoniopropionate (DMSP) wherein said plant is not Panicum commutatum.

2. The method of claim 1 , wherein said plant part is not an isolated plant part and the method is not carried out in vitro.

3. The method of claim 1 or 2, wherein the formulation is a liquid formulation.

4. The method of any preceding claim, wherein contacting the plant or part thereof comprises topical application, coating or watering.

5. The method of claim 4, wherein topical application comprises foliar feeding.

6. The method of claim 5, wherein foliar feeding comprises spraying.

7. The method of claim 4, wherein watering the plant comprises fertigation, hand watering, soil drenching and irrigation.

8. The method of any preceding claim, wherein said plant is a dicot or monocot plant.

9. The method of claim 8, wherein said dicot or monocot plant is selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco, grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut, barley or oats.

10. The method of any preceding claim, wherein the stress is biotic or abiotic stress.

11. The method of claim 10 wherein said abiotic stress is selected from osmotic stress, oxidative stress, heavy metal stress, drought, submergence, cold, UV radiation, heat, ozone, nutrient deficiency and / or salt stress.

12. A genetically altered plant wherein said plant expresses a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90%sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

13. The genetically altered plant of claim 12 wherein said plant expresses a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP- amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acidsequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

14. The genetically altered plant of claim 12 or 13 wherein the plant as defined in a. further expresses an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a MmtN polypeptide comprising SEQ ID NO: 18 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurB polypeptide comprising SEQ ID NO:19 or a functional variant thereof having at least 90% sequence identity thereto or a polypeptide selected from Table 1 b or a functional variant thereof having at least 90% sequence identity thereto.

15. The genetically altered plant of claim 14, wherein said nucleic acid sequences form part of one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve nucleic acid constructs.

16. The genetically altered plant of any one of claims 12 to 15, wherein the nucleic acid encoding a SaSDC polypeptide comprises SEQ ID NO:1 or a functional variant thereof having at least 90% sequence identity thereto.

17. The genetically altered plant of any one of claims 12 to 16, wherein the nucleic acid encoding a SaDOX polypeptide comprises SEQ ID NO:3 or a functional variant thereof having at least 90% sequence identity thereto.

18. The genetically altered plant of any one of claims 12 to 17, wherein the nucleic acid encoding a SaMMTI polypeptide comprises SEQ ID NO:5 or a functional variant thereof having at least 90% sequence identity thereto.

19. The genetically altered plant of any one of claims 12 to 18, wherein the nucleic acid encoding a Burl polypeptide comprises SEQ ID NO: 20 or a functional variant thereof having at least 90% sequence identity thereto.

20. The genetically altered plant of any one of claims 12 to 19, wherein the nucleic acid encoding a BurD polypeptide comprises SEQ ID NO: 21 or a functional variant thereof having at least 90% sequence identity thereto.21 . The genetically altered plant of any one of claims 12 to 20, wherein the nucleic acid encoding a DsyGD polypeptide comprises SEQ ID NO: 22 or a functional variant thereof having at least 90% sequence identity thereto.

22. The genetically altered plant of any one of claims 12 to 21 , wherein the nucleic acid encoding a DSYE polypeptide comprises SEQ ID NO: 23 or a functional variant thereof having at least 90% sequence identity thereto.

23. The genetically altered plant of any one of claims 12 to 22, wherein the nucleic acid encoding a DsyB polypeptide comprises SEQ ID NO: 24 or a functional variant thereof having at least 90% sequence identity thereto.

24. The genetically altered plant of any one of claims 12 to 23, wherein the nucleic acid encoding a DSYB polypeptide comprises SEQ ID NO: 25 or a functional variant thereof having at least 90% sequence identity thereto.

25. The genetically altered plant of any one of claims 12 to 24, wherein the nucleic acid encoding a TpMMT polypeptide comprises SEQ ID NO: 26 or a functional variant thereof having at least 90% sequence identity thereto.

26. The genetically altered plant of any one of claims 12 to 25, wherein the nucleic acid encoding a MmtN polypeptide comprises SEQ ID NO: 27 or a functional variant thereof having at least 90% sequence identity thereto.

27. The genetically altered plant of any one of claims 12 to 26, wherein the nucleic acid encoding a BurB polypeptide comprises SEQ ID NO: 28 or a functional variant thereof having at least 90% sequence identity thereto.

28. The genetically altered plant of any of claims 12 to 27, wherein said plant overexpresses the SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence.

29. The genetically altered plant of claim 28, wherein the nucleic acid construct comprising a nucleic acid sequence encoding a SaSDC, SaDOX, SaMMT 1 , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB polypeptide and further comprises a constitutive promoter.

30. The genetically altered plant of claim 29 wherein said promoter is selected from actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promoter, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfa H3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V-ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters.31 . The genetically altered plant of any one of claims 12 to 30 wherein the exogenous nucleic acid construct is stably incorporated into the plant genome.

32. The genetically altered plant of any one of claims 12 to 31 wherein the exogenous nucleic acid construct is inserted into the plant genome using targeted genome modification.

33. The genetically altered plant of claim 32 wherein the nucleic acid construct is inserted into the plant genome using a rare-cutting endonuclease, for example a TALEN, ZFN or CRISPR / Cas9.

34. The genetically altered plant of any one of claims 12 to 33, wherein said plant is a dicot or monocot plant.

35. The genetically altered plant of claim 34, wherein said dicot or monocot plant is selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco,grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut, barley or oats.

36. The genetically altered plant of any one of claims 12 to 35, wherein the plant has an altered stress response compared to a control plant.

37. The genetically altered plant of claim 36, wherein the stress is abiotic or biotic stress.

38. The genetically altered plant of claim 37 wherein said abiotic stress is selected from osmotic stress, oxidative stress, heavy metal stress, drought, submergence, cold, UV radiation, heat, ozone, nutrient deficiency and / or salt stress.

39. An isolated polypeptide comprising SEQ ID NO: 2 or a sequence having 90% sequence identity thereto.

40. An isolated polypeptide comprising SEQ ID NO: 4 or a sequence having 92% sequence identity thereto.41 . An isolated polypeptide comprising SEQ ID NO: 6 or a sequence having 90% sequence identity thereto.

42. An isolated nucleic acid sequence comprising SEQ ID NO: 1 or a sequence having 90% sequence identity thereto.

43. An isolated nucleic acid sequence comprising SEQ ID NO: 3 or a sequence having 92% sequence identity thereto.

44. An isolated nucleic acid sequence comprising SEQ ID NO: 5 or a sequence having 90% sequence identity thereto.

45. A vector comprising an isolated nucleic acid sequence of any of claims 42 to 44.

46. The vector of claim 45 further comprising a promoter sequence operably linked to said nucleic acid sequence.

47. A host cell comprising an isolated nucleic acid sequence of any of claims 42 to 44 or the vector of claim 45 or 46.

48. The use of a nucleic acid sequence of any of claims 42 to 44 or the vector of claim 45 or 46 for altering the stress response of a plant.

49. A method for altering the stress response of a plant, comprising introducing into said plant a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burlpolypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

50. The method of claim 49 comprising introducing into said plant a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ IDNO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto.51 . The method of a. of claim 49 or 50 wherein said method further comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a MmtN polypeptide comprising SEQ ID NO: 18 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurB polypeptide comprising SEQ ID NO:19 or a functional variant thereof having at least 90% sequence identity thereto or a polypeptide selected from Table 1 b or a functional variant thereof having at least 90% sequence identity thereto.

52. The method of any one of claims 49 to 51 , wherein said nucleic acid sequences form part of one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve nucleic acid constructs.

53. The method of any one of claims 49 to 52, wherein the nucleic acid encoding a SaSDC polypeptide comprises SEQ ID NO:1 or a functional variant thereof having at least 90% sequence identity thereto.

54. The method of any one of claims 49 to 53, wherein the nucleic acid encoding a SaDOX polypeptide comprises SEQ ID NO:3 or a functional variant thereof having at least 90% sequence identity thereto.

55. The method of any one of claims 49 to 54, wherein the nucleic acid encoding a SaMMT 1 polypeptide comprises SEQ ID NO:5 or a functional variant thereof having at least 90% sequence identity thereto.

56. The method of any one of claims 49 to 55, wherein the nucleic acid encoding a Burl polypeptide comprises SEQ ID NO: 20 or a functional variant thereof having at least 90% sequence identity thereto.

57. The method of any one of claims 49 to 56, wherein the nucleic acid encoding a BurD polypeptide comprises SEQ ID NO: 21 or a functional variant thereof having at least 90% sequence identity thereto.

58. The method of any one of claims 49 to 57, wherein the nucleic acid encoding a DsyGD polypeptide comprises SEQ ID NO: 22 or a functional variant thereof having at least 90% sequence identity thereto.

59. The method of any one of claims 49 to 58, wherein the nucleic acid encoding a DSYE polypeptide comprises SEQ ID NO: 23 or a functional variant thereof having at least 90% sequence identity thereto.

60. The method of any one of claims 49 to 59, wherein the nucleic acid encoding a DsyB polypeptide comprises SEQ ID NO: 24 or a functional variant thereof having at least 90% sequence identity thereto.61 . The method of any one of claims 49 to 60, wherein the nucleic acid encoding a DSYB polypeptide comprises SEQ ID NO: 25 or a functional variant thereof having at least 90% sequence identity thereto.

62. The method of any one of claims 49 to 61 , wherein the nucleic acid encoding a TpMMT polypeptide comprises SEQ ID NO: 26 or a functional variant thereof having at least 90% sequence identity thereto.

63. The method of any one of claims 49 to 62, wherein the nucleic acid encoding a MmtN polypeptide comprises SEQ ID NO: 27 or a functional variant thereof having at least 90% sequence identity thereto.

64. The method of any one of claims 49 to 63, wherein the nucleic acid encoding a BurB polypeptide comprises SEQ ID NO: 28 or a functional variant thereof having at least 90% sequence identity thereto.

65. The method of any one of claims 49 to 64, wherein said plant overexpresses an exogenous SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence.

66. The method of claim 65, wherein the nucleic acid construct comprising a nucleic acid sequence encoding a SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB polypeptide further comprises a constitutive promoter.

67. The method of claim 66 wherein said promoter is selected from actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promoter, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfa H3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V-ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters.

68. The method of any of claims 49 to 67 wherein the exogenous nucleic acid construct is stably incorporated into the plant genome.

69. The method of any of claims 49 to 68 wherein the exogenous nucleic acid construct is inserted into the plant genome using targeted genome modification.

70. The method of claim 65 wherein the nucleic acid construct is inserted into the plant genome using a rare-cutting endonuclease, for example a TALEN, ZFN or CRISPR / Cas9.71 . The method of any of claims 49 to 70, wherein said plant is a dicot or monocot plant.

72. The method of claim 71 , wherein said dicot or monocot plant is selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco, grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut, barley or oats.

73. The method of any of claims 49 to 72, wherein the stress is abiotic or biotic stress.

74. The method of claim 73 wherein the abiotic stress is selected from osmotic stress, oxidative stress, heavy metal stress, drought, submergence, cold, UV radiation, heat, ozone, nutrient deficiency and / or salt stress.

75. A method for producing a plant with an altered stress response, comprising introducing into said plant a. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD or Burl polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DysB, TpMMT, DSYE, DysGD, DSYB or DsyG polypeptide comprising a sequence as listed in Table 1 b or a functional variant thereof having at least 90% sequence identity thereto; and / or c. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding an enzyme involved in the conversion of 3-methylthiopropylamine to 3-methylmercaptopropionate.

76. The method of claim 75 comprising introducing into said plant a. an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a S- methylmethionine decarboxylase (SaSDC) polypeptide comprising SEQ ID NO:2 or a functionalvariant thereof having at least 90% sequence identity thereto and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a Burl polypeptide comprising SEQ ID NO: 11 or a functional variant thereof having at least 90% sequence identity thereto, and expressing an exogenous nucleic acid construct comprising an nucleic acid sequence encoding a DMSP-amine oxidase (SaDOX) polypeptide comprising SEQ ID NO:4 or a functional variant thereof having at least 90% sequence identity thereto, and / or expressing an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurD polypeptide comprising SEQ ID NO: 12 or a functional variant thereof having at least 90% sequence identity thereto; and / or b. an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyGD polypeptide comprising SEQ ID NO: 13 or a functional variant thereof having at least 90% sequence identity thereto, and an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYE polypeptide comprising SEQ ID NO: 14 or 31 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DsyB polypeptide comprising SEQ ID NO: 15 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a DSYB polypeptide comprising SEQ ID NO: 16 or 29 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a TpMMT polypeptide comprising SEQ ID NO: 17 or a functional variant thereof having at least 90% sequence identity thereto.

77. The method of a. of claim 75 or 76 wherein said method further comprises introducing into said plant an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a methionine S-methyltransferase (SaMMTI) polypeptide comprising SEQ ID NO:6 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a MmtN polypeptide comprising SEQ ID NO:18 or a functional variant thereof having at least 90% sequence identity thereto, and / or an exogenous nucleic acid construct comprising a nucleic acid sequence encoding a BurB polypeptide comprising SEQ ID NO:19 or a functional variant thereof having at least 90% sequence identity thereto.

78. The method of any one of claims 75 to 77, wherein said nucleic acid sequences form part of one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve nucleic acid constructs.

79. The method of any one of claims 75 to 78, wherein the nucleic acid encoding a SaSDC polypeptide comprises SEQ ID NO:1 or a functional variant thereof having at least 90% sequence identity thereto.

80. The method of any one of claims 75 to 79, wherein the nucleic acid encoding a SaDOX polypeptide comprises SEQ ID NO:3 or a functional variant thereof having at least 90% sequence identity thereto.81 . The method of any one of claims 75 to 80, wherein the nucleic acid encoding a SaMMT 1 polypeptide comprises SEQ ID NO:5 or a functional variant thereof having at least 90% sequence identity thereto.

82. The method of any one of claims 75 to 81 , wherein the nucleic acid encoding a Burl polypeptide comprises SEQ ID NO: 20 or a functional variant thereof having at least 90% sequence identity thereto.

83. The method of any one of claims 75 to 82, wherein the nucleic acid encoding a BurD polypeptide comprises SEQ ID NO: 21 or a functional variant thereof having at least 90% sequence identity thereto.

84. The method of any one of claims 75 to 83, wherein the nucleic acid encoding a DsyGD polypeptide comprises SEQ ID NO: 22 or a functional variant thereof having at least 90% sequence identity thereto.

85. The method of any one of claims 75 to 84, wherein the nucleic acid encoding a DSYE polypeptide comprises SEQ ID NO: 23 or a functional variant thereof having at least 90% sequence identity thereto.

86. The method of any one of claims 75 to 85, wherein the nucleic acid encoding a DsyB polypeptide comprises SEQ ID NO: 24 or a functional variant thereof having at least 90% sequence identity thereto.

87. The method of any one of claims 75 to 86, wherein the nucleic acid encoding a DSYB polypeptide comprises SEQ ID NO: 25 or a functional variant thereof having at least 90% sequence identity thereto.

88. The method of any one of claims 75 to 87, wherein the nucleic acid encoding a TpMMT polypeptide comprises SEQ ID NO: 26 or a functional variant thereof having at least 90% sequence identity thereto.

89. The method of any one of claims 75 to 88, wherein the nucleic acid encoding a MmtN polypeptide comprises SEQ ID NO: 27 or a functional variant thereof having at least 90% sequence identity thereto.

90. The method of any one of claims 75 to 89, wherein the nucleic acid encoding a BurB polypeptide comprises SEQ ID NO: 28 or a functional variant thereof having at least 90% sequence identity thereto.91 . The method of any one of claims 75 to 90, wherein said plant overexpresses an exogenous SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB nucleic acid sequence.

92. The method of claim 91 , wherein the nucleic acid construct comprising a nucleic acid sequence encoding a SaSDC, SaDOX, SaMMTI , Burl, BurD, DsyGD, DSYE, DsyB, DSYB, TpMMT, MmtN and / or BurB polypeptide further comprises a constitutive promoter.

93. The method of claim 92 wherein said promoter is selected from actin, Act2, Act1 , pUbi, Ubi.U4, Ubil promoter, HMGP, CaMV19S, CaMV 35S, GOS2, rice cyclophilin, maize H3 histone, alfalfaH3 histone, 34S FMV, rubisco small subunit, OCS, SAD1 , SAD2, nos, V-ATPase, super promoter, BSV, CsVMV, MMV, FMV G-box proteins and synthetic promoters.

94. The method of any of claims 75 to 93 wherein the exogenous nucleic acid construct is stably incorporated into the plant genome.

95. The method of any of claims 75 to 94 wherein the exogenous nucleic acid construct is inserted into the plant genome using targeted genome modification.

96. The method of claim 95 wherein the nucleic acid construct is inserted into the plant genome using a rare-cutting endonuclease, for example a TALEN, ZFN or CRISPR / Cas9.

97. The method of any of claims 75 to 96, wherein said plant is a dicot or monocot plant.

98. The method of claim 97, wherein said dicot or monocot plant is selected from rice, maize, wheat, sorghum, brassica, soybean, cotton, millet, tomato, potato, pepper, tobacco, grapevine, cucumber, citrus, apple, strawberry, cassava, thale cress, pecan trees, peas, beans, broad beans, lentils, chickpea, cowpea, alfalfa, peanut barley or oats.

99. The method of any of claims 75 to 98, wherein the stress is abiotic or biotic stress.

100. The method of claim 99 wherein said abiotic stress is selected from osmotic stress, oxidative stress, heavy metal stress, biotic stress, drought, cold, UV radiation, heat, ozone, nutrient deficiency, submergence and / or salt stress.

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