Methods of improving disease resistance in plants

Abstract

The invention relates to method of improving disease resistance, an in particular fungal and oomycete disease resistance, in plants. Also described are genetically altered plants characterised by the above phenotype as well as methods of producing such plants.

Description

[0001] M& C PC932446GB

[0002] 1

[0003] Methods of improving disease resistance in plants

[0004] FIELD OF THE INVENTION

[0005] The invention relates to method of improving plant disease resistance, and in particular disease resistance caused by fungal and oomycete pathogens. Also described are genetically altered plants characterised by the above phenotype as well as methods of producing such plants.

[0006] BACKGROUND OF THE INVENTION

[0007] After many years of intense research, plant diseases continue to cause severe damage. The major crops suffer more than 20% yield losses globally despite pathogen control measures being taken. Disease resistance (R)-genes have been identified for many pathogens, but when deployed in crops, they are typically only efficient for a few years until the pathogen evolves new genotypes that overcome the resistance. As such alternative types of plant disease resistance are much needed.

[0008] Most classical R-genes encode nucleotide-binding leucine-rich repeat (NLR) intracellular receptors. They function according to the zig-zag model describing that plant plasma membrane (PM) pattern recognition receptor (PRR) kinases initially recognize conserved pathogen molecular patterns, whereby pattern triggered-immunity (PTI) is activated. Pathogens secrete effector proteins to the plant cell to target and inhibit PTI components. Meanwhile, plants have a large number of NLRs, and occasionally single NLRs are able to directly or indirectly detect effectors and activate effector-triggered immunity (ETI) in which case they are referred to as R-proteins, encoded by R-genes. While PTI and ETI involve transcriptional reprogramming, where many plant genes are up-regulated, ETI as well include the hypersensitive response (HR) programmed cell death that mediates efficient immunity to biotrophic pathogens. NLRs in dicot plants appear in three types according to their N-terminal domains, the Coiled-coil-type sensor NLRs (CNLs), the Toll / interleukin-1-type sensor NLRs (TNLs), and the Resistance-to-powdery-mildew8-type helper NLRs (RNLs). Arabidopsis has -200 NLRs of which -125 are TNLs, and it has been demonstrated that resistance mediated by TNLs depend on Enhanced Disease Susceptibilityl (EDS1). TNLs have recently been demonstrated to activate HR by their intrinsic NADase activity, generating ADP-ribose and cyclicADP-ribose, which in turnM& C PC932446GB

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[0010] activates a heterodimer of EDS1 and Senescence-Associated Carboxylesterase 101 (SAG101) to pass on the signal to RNLs that triggers the HR. At the same time another heterodimer of EDS1 and PhytoAlexin Deficient 4 (PAD4) is formed, which enters the nucleus to activate an immunity transcriptome by help of other RNLs. Newly, this TNL / EDS1 / RNL immunity activation system has in addition been demonstrated to function in PTI downstream of PRR activation. As a result, eds1 mutant plants are severely immune-hampered.

[0011] As an alternative to using naturally occurring R-genes for disease control, mutant screens have been made to search for loss of susceptibility genes, so-called S-genes. Here many recessive mutations that confer resistance have been identified. However, such mutations generally do not affect genuine susceptibility mechanisms required for pathogen growth. Rather, the S-proteins are negative regulators of immunity, causing the homozygous mutant plants to become autoimmune and therefore resistant to more pathogens. Examples are Arabidopsis pmr4, dmr6, edr1 and edr2. Alternatively, primary metabolites may accumulate to levels toxic to the pathogen. Examples are Arabidopsis rsp1, rsp2 and dmr1.

[0012] The most prominent example of an S-gene is Mildew Locus O (MLO), where recessive knock-out mutations confer resistance to powdery mildew. The first barley mlo mutant line was described in 1942, and many have been discovered later. Today mlo resistance is used broadly in barley cultivation, and knocking out orthologues in several plant species has proven to give powdery mildew resistance as well. In Arabidopsis, MLO2, MLO6 and MLO12 are partly redundant. Yet, mlo2 mutations cause some level of resistance, whereas mlo2 mlo6 mlo12 triple mutants are strongly resistant. Importantly, mlo plant genotypes often suffer from pleiotropic effects including leaf lesions and growth retardation.

[0013] Thus, there exists a need to develop new methods to improve disease resistance in plants. The present invention addresses this need.

[0014] SUMMARY OF THE INVENTION

[0015] Plant diseases continue to cause severe damages in crops and to alleviate this problem we identified a novel disease resistance mechanism. To identify new disease resistantM& C PC932446GB

[0016] 3

[0017] mutants with less growth retarding autoimmunity, we used an EMS mutant population in the immune-compromised eds1-2 genetic background. We identified an allelic series of recessive immunela (immla) mutations that provide resistance to the powdery mildew fungus, Golovinomyces orontii (Go). This EDS1 -independent resistance is partly manifested at the stage of penetration when papilla formation is stronger, and partly after penetration when imm1 is causing shorter fungal hyphae. IMM1a is identical to the sulfate transporter, SULTR3;3 (and such terms are used interchangeably herein), and the resistance phenotype shown in the mutants can be complemented by overexpression of the otherwise poorly expressed SULTR3;4 (=IMM1b) (again, such terms are used interchangeably). This indicates that the two proteins share a function in making a plant susceptible to infection.

[0018] Immla mutations also enhance the resistance to two unrelated fungal pathogens, Colletotrichum higginsianum and Botrytis cinerea, as well as to the oomycete pathogen Hyaloperonospora arabidosidis, demonstrating that what we have discovered is a broadspectrum disease resistance pathway. Interestingly, mutations in the closest homologues of IMM1a in barley and rice cause low phytic acid levels. These genes are thus named are low phytic acidl (LPA1) and low phytic acid (LPA), respectively. Thus, imm1a / lpa1 mutations may improve plant disease resistance at the same time as reducing eutrophication of the environment and consequently increasing the nutritional value of grains.

[0019] Accordingly, in a first aspect of the invention there is provided a genetically altered plant, plant part thereof or plant cell, wherein said plant has reduced expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and 3;4, wherein SULTR3;3 is defined in any one of SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, or 54-56 or a functional variant or homolog thereof, and SULTR3;4 is defined in any one of SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, or 56 or a functional variant or homolog thereof, wherein where the sulfate transporter is expressed as more than one homeologue, all homeologues are mutated; wherein said plant is not Arabidopsis.

[0020] In an embodiment, the genetically altered plant, plant part thereof or plant cell comprises at least one mutation in at least one gene encoding a SULTR3;3 protein and / or at least one mutation in the SULTR3;3 promoter, and further at least one mutation in at least one gene encoding a SULTR3;4 protein and / or at least one mutation in the SULTR3;4M& C PC932446GB

[0021] 4

[0022] promoter, wherein said mutation results in reduced expression and / or activity of the sulfate transporter compared to a wild-type or control plant.

[0023] In an embodiment, the mutation is a loss of function mutation or a partial loss of function mutation.

[0024] In another embodiment, the genetically altered plant, plant part thereof or plant cell comprises at least one RNAi construct, wherein the RNAi construct reduces the expression of the SULTR3;3 and SULTR3;4 gene compared to a wild-type or control plant.

[0025] In an embodiment, the genetically altered plant, plant part thereof has increased fungal disease resistance compared to a wild-type or control plant. Preferably, the genetically altered plant, plant part thereof has increased resistance to at least one of Golovinomyces orontii, Colletotrichum higginsianum, Botrytis cinerea.

[0026] In an embodiment, the genetically altered plant, plant part thereof or plant cell is a monocot or a dicot. Preferably the plant is a crop plant. In an embodiment, the plant is selected from wheat, maize, rice, barley, rye, oat millet, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, carrot, parsnip, turnip, zucchini, pumpkin, cucumber, tomato, watermelon, melon, zucchini, squash, pumpkin, gourd, pea, faba bean, common bean, soybean, chickpea, sugarbeet, grape, blueberry, sugar cane and ryegrass.

[0027] In an embodiment, plant growth or yield is not affected by the genetic alteration.

[0028] In an embodiment, the genetically altered plant, plant part thereof is characterised by reduced phytic acid levels compared to a wild-type or control plant.

[0029] In an embodiment, the plant part is a seed or grain.

[0030] In a further aspect of the invention, there is provided a genetically altered plant, plant part thereof or plant cell, wherein said plant is characterised by reduced expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3, wherein where the sulfate transporter is expressed as more than one homeologue in the plant, part thereof or plant cell, all homeologues are mutated, wherein the plant is tomato.

[0031] In one embodiment, the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NO: 39 or 40 or a functional variant or homolog thereof. Preferably, the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NO: 39 or a functional variant or homolog thereof.M& C PC932446GB

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[0033] In a further aspect of the invention there is provided the use of the genetically altered plant or plant part thereof of the invention as a food or feedstuff.

[0034] In another aspect of the invention, there is method of increasing disease resistance in a plant, plant part thereof or plant cell wherein said method comprises reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4 compared to a wild-type sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4.

[0035] In another aspect of the invention, there is provided a method of producing a plant with increased resistance to disease, the method comprising reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4, compared to a wild-type sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4.

[0036] In an embodiment, SULTR3;3 comprises an amino acid sequence as defined in one of SEQ ID NO: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56 or a functional variant or homolog thereof and / or SULTR3;4 comprises an amino acid sequence as defined in one of SEQ ID NO: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 or a functional variant or homolog thereof.

[0037] In an embodiment, the method comprises increasing resistance to at least one fungal disease and / or an oomycete pathogen. Preferably, the at least one fungal disease is selected from resistance to at least one of Podosphaera spp, Leveillula spp, Sphaerotheca spp., Blumeria spp, Erysiphe spp, Golovinomyces orontii, Colletotrichum higginsianum, Botrytis cinerea, or Blumeria spp.

[0038] In an embodiment, the method comprises introducing at least one mutation into at least one gene encoding a SULTR3;3 protein and / or introducing at least one mutation in the SULTR3;3 promoter, wherein said mutation results in reduced expression and / or activity of the sulfate transporter compared to a wild-type or control plant.

[0039] In an embodiment, the method comprises introducing at least one mutation into at least one gene encoding a SULTR3;4 protein and / or introducing at least one mutation in the SULTR3;4 promoter, wherein said mutation results in reduced expression and / or activity of the sulfate transporter compared to a wild-type or control plant.

[0040] In another embodiment, the method comprises introducing at least one mutation into at least one gene encoding a SULTR3;3 protein and / or introducing at least one mutation in the SULTR3;3 promoter, and further at least one mutation into at least one geneM& C PC932446GB

[0041] 6

[0042] encoding a SULTR3;4 protein and / or introducing at least one mutation in the SULTR3;4 promoter, wherein said mutation results in reduced expression and / or activity of the sulfate transporter compared to a wild-type or control plant.

[0043] In an embodiment, where the sulfate transporter is expressed as more than one homeologue, all homeologues are mutated.

[0044] In an embodiment, the mutation is a loss of function mutation or a partial loss of function mutation.

[0045] In an alternative embodiment, the method comprises introducing and expressing in the plant, plant part thereof or plant cell at least one RNAi construct, wherein the RNAi construct reduces the expression of the SULTR3;3 and / or the SULTR3;4 gene.

[0046] In an embodiment, the plant, plant part thereof or plant cell is a monocot or a dicot. Preferably the plant is a crop plant. In an embodiment, the plant is selected from, wheat, maize, rice, barley, rye, oat millet, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, carrot, parsnip, turnip, zucchini, pumpkin, cucumber, watermelon, melon, zucchini, squash, pumpkin, gourd, pea, faba bean, common bean, soybean, chickpea, soybean, pea, sugarbeet, grape, sunflower, rye, blueberry, sugar cane and ryegrass.

[0047] In an embodiment, the method does not affect plant growth or yield compared to a wildtype or control plant.

[0048] In an embodiment, the method further reduces phytic acid levels compared to a wildtype or control plant.

[0049] In another aspect of the invention, there is provided a method of screening a population of plants and identifying and / or selecting a plant that will have increased disease resistance compared to a control or wild-type plant, the method comprising detecting at least one polymorphism or mutation in the SULTR3;3 and / or 3;4 gene and / or promoter and selecting said plant. Preferably, the polymorphism or mutation is a loss or partial loss of function mutation.

[0050] DESCRIPTION OF THE FIGURES

[0051] The invention is further described in the following non-limiting figures:M& C PC932446GB

[0052] 7

[0053] Figure 1. immla mutants have EDS1 -independent resistance to the powdery mildew fungus, Golovinomyces orontii (Go). A, Disease development (see white cover, exemplified at arrows) of 4-week-old plants inoculated and imaged 10 days later. B, Disease severity index (DSI) visually scored on a scale from 0 to 5 (based on direct visual scoring of disease symptoms of plants imaged in Fig. 2). C, Biomass quantification of Go 7 days after inoculation by qPCR on fungal DNA relative to plant DNA using two biological replicates. Error bars, standard error (SE); *, P<0.05; **, P<0.01; ***, P<0.001 and ****, P<0.0001 determined by one-way ANOVA. D, IMM1a localizes to the plasma membrane in complemented transgenic lines (see A). H, haustorium. Arrow, plasma membrane. imm1 = immla in this figure.

[0054] Figure 2. Golovinomyces orontii disease development on immla mutants and associated lines. Disease development (see white cover, exemplified at arrows) of 4-week-old plants inoculated with Go and imaged 10 days later. Image size and colour settings are not to scale in all cases. imm1 = immla in this figure.

[0055] Figure 3. imm1-resistance to Golovinomyces orontii is conferred by a combination of pre- and post-invasive immunity, the latter neither conferred by HR nor by encasements. A, Summary of callose intensities at 50 attack sites with scores ‘O’, ‘1’, ‘2’, ‘3’ and ‘4’ at 18 hai. B, Penetration rate assessed. C, Average hyphal length developed from attacking spores. D, HR rate assessed. E, encasement rate. F. Image of wildtype (Col-O) infected with powdery mildew and immla resistant to infection. B to E assessed from 100 attack sites 36 hai. n=4. Error bars, SE; *, P<0.05; ***, P<0.001; and ****, P<0.0001 determined by T-test.

[0056] Figure 4. immla shows resistance to the hemibiotrophic fungal pathogen, Colletotricum higginsianum. A, Representative symptoms at 6 dpi. B, Disease severity index (DSI) at 6 dpi (n=7). C, Fungal biomass quantification by qPCR of fungal and Arabidopsis genomic DNA at 3 dpi (n=3). Error bars, SE; *, P<0.05; **, P<0.01; ***, P<0.001 and ****, P<0.0001 determined by one-way ANOVA.

[0057] Figure 5. immla shows resistance to the necrotrophic fungal pathogen, Botrytis cinerea. A, Representative symptoms at 5 dpi. B, Disease severity index at 5 dpi (n=63-69). C, Fungal biomass quantification by qPCR of fungal and Arabidopsis genomic DNA at 3 dpiM& C PC932446GB

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[0059] (n=3). Error bars, SE; *, P<0.05; **, P<0.01; and ***, P<0.001 determined by one-way ANOVA.

[0060] Figure 6. immla shows resistance to the oomycete pathogen, Hyaloperonospora arabidosidis.

[0061] Figure 7. immla plants have normal growth and seed-set. A, Plant growth compared to controls. B, Seed production by plants in A (n=5). Error bars, SE; *, P<0.05; **, P<0.01 determined by one-way ANOVA.

[0062] Figure 8. Eleven immla EMS alleles identified in mutant screen for resistance to Go in the Col-0 eds1-2 genetic background. Several immla alleles found more than once.

[0063] Figure 9. Dose-dependent requirement for AtIMMIa in Go susceptibility. AtIMMIa levels based on GFP-signal intensity visually scored by confocal microscopy and susceptibility levels visually scored as Go symptoms. Result based on 40 GFP-AtlMM1a and 50 AtIMM1a-GFP T1-plants.

[0064] Figure 10. Knock-out of IMM1a (SULTR3;3) is sufficient for obtaining resistance to Golovinomyces orontii, but over-expression of the close relative, SULTR3;4 (IMM1b), complements immla. A, Go development on different sultr3 mutants and combinations hereof. B, IMM1a over-expression reverts the sultr3 quintuble mutant to become susceptible to Go. C, SULTR3;4 (IMM1b) over-expression reverts the sultr3 quintuble mutant and immla to become susceptible to Go.

[0065] Figure 11. IMM1a and SULTR3;4 (IMM1b) are closely related to barley and rice proteins required for seed phytic acid. A, Phylogeny of Arabidopsis so-called sulfate transporters and selected barley and rice homologues. B, Phylogeny of selected plant sulfate transporters.

[0066] Figure 12. imm1-resistance in Arabidopsis can be complemented by barley HvIMMIa. Knock-out of IMM1A=SULTR3;3 (highly expr.), but not of IMM1B=SULTR3;4 (poorly expr.) causes resistance. Yet, the double immla immlb knock-out has marginally improved resistance. Over-expression of the barley IMM1A (LPA1) can complement Arabidopsis imm1A.M& C PC932446GB

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[0068] Figure 13. Schematic of working theory behind pre- and post-invasive immunity. At the microscopic level, immunity to powdery mildew fungi is manifested at three stages. 1) A strong plant epidermal cell wall apposition (papilla) formed at the site of attack can efficiently stop fungal penetration. 2) The fungal haustorium formed in the plant cell after penetration can become enclosed by an extension of the papilla cell wall apposition, referred to as an encasement. 3) The epidermal cell can undergo a programmed cell death, a so-called hypersensitive reaction (HR), which efficiently stops attack by the biotrophic powdery mildew fungi.

[0069] Figure 14. CRISPR / Cas9-generated barley imm1A-2

[0070]

[0071] double mutant line. A CRISPR / Cas9 construct with a single guide RNA (5’-GAGGCTGGCAATGGTGAGGC-3’ part of SEQ ID NO: 73, SEQ ID NO: 87 is full sequence) was transformed into barley cultivar Golden Promise immature embryos using Agrobacterium. After plant regeneration from callus cultures, the Hvimm1A-2 Hvimm1B-1 double mutant was selected. A-B, The Hvimm1A-2 single base insert allele was numbered to follow the Ipa1- 1 allele of this gene in cultivar Harrington (Ye et al., 2011), while the single base insert allele Hvimm1B-1 was numbered to follow the spdt-1 and -2 alleles of this gene in cultivar Golden Promise (Gu et al., 2022). FP and RP, forward and reverse primers, respectively. C, PCR-based demonstration that Hvimm1A-2 Hvimm1B-1 is transgene-free.

[0072] Figure 15. Barley double mutant, Hvimm1A-2 Hvimm1B-1, but not the single mutant Hvimm1A-1, has powdery mildew resistance. A, Top row: Hvimm1A-1 mutant in Harrington background (=low phytic acidl (Ipa1-1)) (Ye et al., 2011). Bottom row; Harrington WT. B, Top row: Hvimm1A-2 Hvimm1B-1 double mutants; bottom row: Golden Promise WT (CRISPR-minus regenerant). C, Quantification of powdery mildew colonies on first leaves of Hvimm1A-1, of mlo (in barley cultivar Ingrid background) and of Hvimm1A-2 Hvimm1B-1, 7 days after inoculation (countings made in boxed areas in A and B). D, Hvimm1A-2 Hvimm1B-1 has pre-invasive immunity to the powdery mildew fungus. Penetration rate quantified 2 days after inoculation from 100 attack sites in each repeat (one leaf). C-D, Error bars, SE; ***, P<0.001 and ****, P<0.0001 determined by T-test. n=3 (C) and n=5 (D).

[0073] Figure 16. Phylogenetic tree and SULTR3;3 and SULTR3;4 sequences. The arrows indicate sequences referred to as SLILTRs, which based on their known functions areM& C PC932446GB

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[0075] predicted cause disease resistance when knocked out or knocked down according to the current invention.

[0076] Figure 17. Knock-out of tomato SIIMM1A, but not of SIIMM1B, causes resistance to the powdery mildew fungus, Oidium neolycopersici. A, Slimm1A-1 mutation in Solyc05g007980 and Slimm1B-1 and -2 mutations in Solyc03g 120250. B-C, Powdery mildew symptoms and colony quantification on leaves of Slimml tomato mutants. T-test: **, P<0.01; ****, P<0.0001. Lack of effect of SI I MM 1B knock-out is likely due to its low transcript expression in leaves.

[0077] DETAILED DESCRIPTION OF THE INVENTION

[0078] The present invention will now be further described. In the following passages, different aspects of the invention are defined in more detail. 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.

[0079] 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.

[0080] 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), natural occurring, mutated, synthetic DNA or RNA molecules, and analogs of the DNA or RNA generated using nucleotide analogs. It can be single-stranded or double-stranded. 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" 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 optionally together with regulatory sequences or may comprise only a coding sequence as in cDNAs, and / or may include cDNAs in combination with regulatory sequences.M& C PC932446GB

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[0082] The terms "polypeptide" and "protein" are used interchangeably herein and refer to amino acids in a polymeric form of any length, linked together by peptide bonds.

[0083] The aspects of the invention involve recombinant DNA technology and exclude embodiments that are solely based on generating plants by traditional breeding methods.

[0084] In an aspect of the invention, there is provided a method of increasing disease resistance in a plant, the method comprising reducing or abolishing the expression and / or activity of a sulfate transporter (SLILTR).

[0085] SLILTRs have traditionally been classified as I / SO42" co-transporters, and are integrated into membranes by 12 membrane-spanning domains, and contain a carboxyl-terminal region, named STAS (Sulfate Transporter / AntiSigma-factor), which is thought to be critical for both activity and stability of the transporters, as well as for their interaction with other proteins.

[0086] Sulfate transporters are encoded by a multigene family. They can be divided into four functional groups based on their amino acid sequence.

[0087] The precise role of group 3 SLILTRs (SLILTR3) has been difficult to establish, in particular as their expression, subcellular localisation and substrate preferences vary. There are five sub-groups within the group 3 SLILTR transporter family, namely SULTR3;1, SULTR3;2, SULTR3;3, SULTR3;4 and SULTR3;5 transporters.

[0088] There is increasing evidence that SULTR3;3 and SULTR3;4 function additionally or alternatively as phosphate transporters. Ding et al. (2020) and Gu et al. (2022) provide evidence that Arabidopsis and barley SULTR3;4 is a phosphate transporter. For example, Gu et al. (2022) refers to HvSPDT - (= barley SULTR3;4) - encoding, a plasma membrane-localized Pi / H+cotransporter, while Ding et al. (2020) reports that AtSPDT - (= AtSULTR3;4) - is a SULTR-like P Distribution Transporter mainly expressed in the vascular cambium.

[0089] SULTR3;3 and SULTR3;4 transporters are also implicated in phytic acid accumulation in grains of barley and rice. As phytic acid is the major storage compound for phosphorous in plants, SULTR3;3 and SULTR3;4 have also been implicated in the transport of phosphorous.M& C PC932446GB

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[0091] Accordingly, in view of the above, the term “SULTR” or “sulfate transporter” used herein refers to an anion transporter. In other words, sulfate transporter in the context of SULTR used herein is not intended to limit the transporter to one that only transports sulfate. In one embodiment, SULTR3;3 and / or SULTR3;4 may be defined as transporters of sulfate and / or of phosphorus-containing anions such as phosphate and / or phytate, and / or of phosphorus-containing compounds (such as phytic acid). That is, a SULTR3;3; and / or SULTR3;4 may be defined as transporting at least sulfate, phosphate and / or phosphorous, and / or a further ion or compound. For the avoidance of doubt, the group 3 SULTRs disclosed herein may be functionally characterised as a H+ / SO42- cotransporter, a phosphorous and / or a phosphate transporter.

[0092] In one embodiment, the group 3 SULTRs is a SULTR3;3. In one embodiment, the transporter is LPA1 in barley and rice (here mutants referred to as immla) or a homolog thereof. For example, Figure 17 shows that a mutation in an SULTR 3;3 ortholog in tomato (SEQ ID NO: 39) is sufficient to confer powdery mildew resistance.

[0093] In one embodiment, the group 3 SULTRs is a SULTR3;4. In one embodiment, the SULTR3;4 transporter is SULTR-like phosphorus distribution transport (SPDT) in barley and rice (here mutants referred to as immlb). Fang et al. (Sci. Adv. 11, eady3442, 2025) provides an insightful structural and functional characterization of the transporter SULTR 3;4, SPDT in rice (OsSPDT). The authors determine that OsSPDT forms a domainswapped homodimer with each protomer comprising a cytoplasmic N-terminal domain (NTD; residues 1-109), a transmembrane domain (TMD; residues 110-519) divided into core and gate subdomains, and containing 14 transmembrane helices (TM1-TM14), and a C-terminal sulfate transporter and antisigma factor (STAS) domain (residues 520-670). The homodimer forms a phosphate-binding pocket, formed by TM1, TM3, TM8, and TM10.

[0094] Interestingly, the authors identify key residues for transporter function and specificity. Within the substrate-binding pocket, a hydrogen-bond network coordinates phosphate via Tyr133from TM1, Ser170from TM3, Glu370from TM8, and Ser413and Ser415from TM10 which are strictly conserved across SULTR orthologs (see fig. S4A of Fang et al). Notably, Arg416from TM10 stabilized the pocket architecture through dual interactions with Ser413side chain and the Phe168backbone carbonyl group, functioning as a structural linchpin conserved in SULTR transporters.M& C PC932446GB

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[0096] The authors of the publication also identified that a residue key for the Pi substrate specificity of OsSPDT was Serine170in TM3. A S170A mutation abolished the phosphate transport function (though had no effect on sulfate transport), establishing Ser170as a necessary but insufficient determinant for anion selectivity. Notably, the author’s phylogenetic analysis demonstrates that Ser170is exclusive to and conserved in the SPDT / SULTR3;4 and SULTR3;3 clades. We concur with this finding - a serine at a position corresponding to OsSPDT is conserved in all the IMM1A and IMM1B proteins in Arabidopsis, barley and tomato described herein.

[0097] Accordingly, in one embodiment, the SULTR3;3 and / or SULTR3;4 or variant thereof of the invention preferably comprises a serine at a position corresponding to position 170 of SEQ ID NO: 7. This is Serine170 of OsSPDT described in Fang et al.

[0098] An alignment with OsSPDT shows that the equivalent residue in Arabidopsis SLILTR 3;3 (IMM1A, SEQ ID NO: 1) and SULTR 3;4 (IMM1B, SEQ ID NO: 2) is Serine124 and Serine147, respectively. Either of SEQ ID NO: 1 or 2 can be used for an alignment to identify the corresponding position of the key residue described in Fang et al. (2025).

[0099] Accordingly, in one embodiment, the SULTR3;3 and / or SULTR3;4 or variant thereof of the invention preferably comprises a serine corresponding to residue 124 of SEQ ID NO: 1 (Arabidopsis IMM1A (SULTR 3;3)) and / or residue 147 of SEQ ID NO: 2 (Arabidopsis (SULT3;4)).

[0100] The corresponding position can be identified by aligning a sequence with the sequence of SEQ ID NO: 1, 2 or 7 (or another suitable sequence) and identifying the conserved SIASL motif - the corresponding residue is the first serine of this motif.

[0101] In Barley, the corresponding residue is residue 167 of SEQ ID NO: 3 (SULTR 3;3) and residue 114 in SEQ ID NO: 4 (SULTR 3;3) and residue 160 of SEQ ID NO: 5 (SULTR 3;4).

[0102] In rice, the corresponding residue is residue 157 in SEQ ID NO: 6 (SULTR 3;3).

[0103] In Tomato, the corresponding residue is residue 151 of SEQ ID NO: 39 (SULTR 3;3) and residue 154 of SEQ ID NO: 40 (SUTR 3;4).M& C PC932446GB

[0104] 14

[0105] In Barley, the corresponding residue is residue 167 of SEQ ID NO: 3 (SULTR 3;3), residue 114 in SEQ ID NO: 4 (SULTR 3;3) and residue 160 of SEQ ID NO: 5 (SULTR 3;4).

[0106] The corresponding residue in SEQ ID NO: 5 is a threonine residue, which is chemically similar to serine (both are polar, uncharged residues). Accordingly, the SULTR3;3 and / or SULTR3;4 or variant thereof of the invention preferably comprises a serine or a threonine corresponding to residue 124 of SEQ ID NO: 1 (Arabidopsis IMM1A (SULTR 3;3)) and / or residue 147 of SEQ ID NO: 2 (Arabidopsis (SULT3;4)). This can be identified by aligning against the SIASL or TIASL motif.

[0107] SULTR3;3 and SULTR3;4 transporters have high overall sequence homology (65% across the entire sequence), with the most sequence variation being seen between the 9thtransmembrane domain and the C-terminus (including the STAS domain). Preferably, the method comprises reducing the expression or activity of a sulfate transporter (SULTR) selected from the subfamily 3;3 and / or 3;4. The method may comprise reducing the expression of SULTR3;3. Alternatively, the method may comprise reducing the expression of SULTR3;4. Alternatively, the method may comprise reducing the expression of SULTR3;3 and 3;4.

[0108] The term “expression” means genes that are transcribed into mRNA and then translated into a protein as well as those that are transcribed into mRNA but are not translated into protein.

[0109] By “disease resistance” is meant the ability to prevent or reduce the presence of diseases in otherwise susceptible hosts i.e., the ability to withstand attack from pathogens. A pathogen is defined in the art as an organism that causes disease to its host, pathogens include viruses, bacteria, oomycetes, protists and fungi.

[0110] Preferably, the method comprises increasing disease resistance to at least one fungal pathogen. As shown in Figure 1 to 6, reducing the expression or activity of a SULTR leads to an increase in disease resistance to a number of different species of fungal pathogens, proving that the mechanism described herein increases plant immunity to any fungal pathogen.M& C PC932446GB

[0111] 15

[0112] In one example, the fungal pathogen may be selected from the phyla Ascomycota and Basidiomycota. Where the pathogen is from the phyla Ascomycetes, the pathogen may be selected from one of the following classes: Dothideomycetes (e.g., Cladosporium spp.), Sordariomycetes (e.g., Magnaporthe spp.), or the Leotiomycetes (e.g., Botrytis spp.). Where the pathogen is from the phyla Basidiomycetes, the pathogen may be selected from one of the following classes: (Pucciniomycetes) and the smuts (spread among the subphylum of Ustilaginomycotina).

[0113] In one example, the fungal pathogen may be selected from Erysiphaceae family (such as, Podosphaera spp.) Leveillula spp, Sphaerotheca spp. Pyricularia oryzae, Botrytis cinerea, Puccinia spp, Fusarium graminearum, Fusarium oxysporum, Blumeria spp., Zymoseptoria tritici, Colletotrichum spp, Ustilago maydis, Melampsora Uni, Phakopsora pachyrhizi, Golovinomyces orontii, Colletotrichum higginsianum, Erysiphe spp. (such as, Erysiphe graminis, Erysiphe cichoracearum, Erysiphe cruciferarum, Erysiphe lycopersici, Erysiphe pisi, Erysiphe heracleid, Erysiphe polygon!, Erysiphe necatorQ, Rhynchosporium graminicola (leaf blotch or scald of barley), Bipolaris spp, Alternaria spp., Sclerotinia spp, Verticillium spp. Pyrenophora spp., Ramularia spp., Pseudocercospora fijiensis (banana sigatoka disease), Cercospora zeina on maize and Rhizoctonia solani.

[0114] Preferably, the fungal pathogen is selected from Podosphaera spp, Leveillula spp, Sphaerotheca spp., Blumeria spp, Erysiphe spp., Golovinomyces orontii, Colletotrichum higginsianum, Botrytis cinerea, Blumeria spp, or, Hyaloperonospora arabidosidis.

[0115] It will be clear to the skilled person that any fungal pathogen whose pathogenicity is impacted by SULTR3;3 and / or SULTR3;4 function will fall within the scope of the invention.

[0116] As shown in Figure 6, the Applicant has demonstrated that the invention can be used to increase resistance against oomycete pathogens, such as the parasitic model pathogen Hyaloperonospora arabidosidis.

[0117] In a particular example, the fungal pathogen may be selected from Golovinomyces orontii, Colletotrichum higginsianum, Botrytis cinerea, Blumeria spp, or, Hyaloperonospora arabidosidis.M& C PC932446GB

[0118] 16

[0119] By “increasing disease resistance” is meant at least a decrease of disease symptoms and / or pathogen quantity by least 50%. There are a number of known techniques in the art to measure fungal resistance. For example, image-based symptom scoring can be used to measure disease resistance. Here, a plant is exposed to a known pathogen and images are taken of said plant over a given time-period. The images are then subjected to extensive analysis using a variety of tools, including but not limited to, pixel classification using colour hue values from RGB images and an algorithm (such as but not limited to a random forest algorithm) to establish necrotic, chlorotic and healthy areas of the plant. Chlorophyll fluorescence imaging can also be used to determine diseased areas and the proportion of the plant that they account for. Resistance can as well be determined by quantifying the amount of pathogen by quantitative PCR of pathogen genomic DNA relative to plant genomic DNA. In addition, resistance can be determined by microscopy by counting and measuring the progression of the fungal structures.

[0120] The methods described herein may also comprise the step of measuring an increase in disease resistance, and in particular, an increase in fungal disease resistance. This may be achieved using known techniques in the art, for example, one of the above-described methods.

[0121] As used herein, the terms “reducing” means a decrease in the levels of expression and / or activity of a sulfate transporter by up to or more than 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% when compared to the level in a wild-type or control plant. In one embodiment reducing means a decrease of at least 50% compared to the level in a wildtype or control plant. Reducing may or may not encompass changes in the absolute sulfate transporter transcript level, preferably it does not. Reducing also may or may not encompass abolishing expression. The term “abolish” expression means that no expression of a sulfate transporter is detectable (no transcript) or that no functional polypeptide is produced. These reductions can be measured by any standard technique known to the skilled person. For example, a reduction in the levels of expression and / or content levels may be a measure of nucleic acid levels and / or protein levels and can be measured by any technique known to the skilled person, such as, but not limited to, any form of quantitative PCR, gel electrophoresis and immunoblotting or chromatography (e.g. HPLC).M& C PC932446GB

[0122] 17

[0123] A reduction in activity may also refer to a complete or partial loss-of-function. Accordingly, the method may result in a complete or partial loss of sulfate, phosphate and / or phosphorous transport by the sulfate transporter. By complete, may mean that no phosphate and / or phosphorous sulfate transport can be detected. Levels of phosphate, phosphorous or sulfate transport can be measured using any known technique in the art, for example, using a medium containing a known amount of sulfate, phosphate and / or phosphorous and using mass spectrometry to quantify sulfate, phosphate and / or phosphorous levels before and after a time period. Alternatively, a FRET based sensor (FLIP-SP) can be used to measure the level of sulfate, phosphate and / or phosphorous inside cells with the level of fluorescence corresponding to the level of sulfate, phosphate and / or phosphorous.

[0124] The method may additionally lead to a decrease in phytic acid levels in the plant. Accordingly, there is provided a method of increasing resistance to disease, preferably fungal disease, and decreasing phytic acid levels in the plant. Phytic acid (PA) is the main phosphorus (P) store in seeds. It cannot be digested by humans and monogastric animals who lack the digestive enzyme phytase. For this reason, almost 90% of phytate consumed by humans is excreted, contributing to eutrophication of rivers, lakes, and oceans. Further, as phytic acid chelates divalent cations of nutritional importance for humans and animals, such as Zn2+ and Fe2+, phytic acid in consumed seed products may cause malnutrition. For this reason, it is desirable to reduce the levels of phytic acid in plants.

[0125] By decrease in phytic acid levels is meant a decrease in phytic acid levels compared to a wild-type or control plant. By decrease is meant, a decrease of at least 2%, 4%, 5%, 6%, 8%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% when compared to the level in a wild-type or control plant. To measure phytic acid levels, a common method is to extract the phytic acid from a sample using an acidic solution, then react it with an enzyme (phytase) to release inorganic phosphorus, which is then quantified using a colorimetric assay based on the formation of molybdenum blue, allowing for calculation of the initial phytic acid concentration based on the released phosphorus amount; this can be done using a commercially available phytic acid assay kit.

[0126] Accordingly, in another aspect of the invention, there is provided a method of improving the nutritional content of a plant, the method comprising reducing the expression and / orM& C PC932446GB

[0127] 18

[0128] activity of a sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4 as described herein. In a preferred embodiment, 'improving the nutritional content of a plant’ is meant decreasing phytic acid level in a plant as described herein.

[0129] There is also provided a method of increasing resistance to disease, preferably fungal disease, and decreasing phytic acid levels in a plant, comprising reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4 as described herein.

[0130] In one embodiment, the method comprises reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3, as described herein. For example, in Figure 17, only a mutation in the SULTR3;3 gene is required to confer powdery mildew resistance.

[0131] In one embodiment, the method comprises reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;4, as described herein.

[0132] In one embodiment, the method comprises reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and SLILTR 3;4, as described herein. For example, Figure 15 shows that in Barley a mutation in each of a SLILTR 3;3 and a SLILTR 3;4 is required to confer powdery mildew resistance.

[0133] There is also provided a method of reducing eutrophication, the method comprising the method comprising reducing the expression and / or activity of a sulfate transporter (SULTR) subfamily 3;3 and / or SULTR3;4.

[0134] It is common for genetically altered plants with broad-range disease resistance to suffer from growth penalties. This limits the commercial application of such plants. However, as shown in Figure 7, the plants of the present invention have no growth penalties. Accordingly, there is provided a method of increasing resistance to disease, preferably fungal disease without any effect on plant growth or yield. There is also provided a method of increasing resistance to disease, preferably fungal disease, and decreasing phytic acid levels in the plant, again without any effect on plant growth or yield.M& C PC932446GB

[0135] 19

[0136] Accordingly, there is provided a method of increasing resistance to disease and increasing plant yield in the plant compared to a wild-type or control plant, the method comprising reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4 as described herein. Preferably, increasing plant yield in a plant means increasing seed or grain number in a plant. As shown in Figure 7, immla mutants show an increased number of seed per plant compared to controls.

[0137] Also provided is a method of increasing resistance to disease, decreasing phytic acid levels and increasing plant yield in the plant compared to a wild-type or control plant, the method comprising reducing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 and / or SULTR3;4 as described herein.

[0138] Plant growth generally refers to biomass accumulation. This is the increase of plant volume and / or mass with or without the formation of new structures such as organs, tissues and cells. Plant growth is typically associated with development and reproduction. Plant growth can be assessed by measuring any one of the following parameters biomass, flowering time, seed production and percentage of seed germination amongst other variables.

[0139] The term yield in general means produce that has economic value, usually related to a specific crop, in an area within a period of time. Individual plant materials directly contribute to yield based on their number, size and / or weight. Alternatively, 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. Yield can be measured through assessing one or more of (a) biomass (weight) of one or more parts of a plant, aboveground (harvestable parts), or root biomass, root volume, root length, root diameter or root length or biomass of any other harvestable part. Biomass may be expressed as g / plant or kg / hectare (b) seed yield per plant, which may comprise one or more of seed biomass (weight) per plant or an individual basis, (c) seed filling rate, (d) number of filled seeds, (e) harvest index, which may be expressed as a ratio of the yield of harvestable parts such as seeds over the total biomass, (f) viability / germination efficiency, (g) number or size or weight of seeds or pods or beans or grain (h) seed volume (which may be a result of a change in the composition (i.e. lipid (also referred to herein as oil)), protein, and carbohydrate total content and composition), (i) (individual or average) seed area, (j) individual or average seed length, (k) individualM& C PC932446GB

[0140] 20

[0141] or average seed width, (I) individual or average seed perimeter, (m) growth or increased branching, for example inflorescences with more branches, (n) fresh weight or grain fill (o) ear weight (p) thousand kernel weight (TKW), which may be taken from the number of filled seeds counted and their total weight, amongst other variables.

[0142] By “no effect” is meant no significant or observable effect on any one of the above growth or yield parameters.

[0143] In one embodiment, the method comprises introducing at least one mutation into the, preferably endogenous, gene encoding SULTR3;3 and / or SULTR3;4 and / or the SULTR3;3 and / or SULTR3;4 promoter. The mutation may be any mutation that reduces or abolishes the phosphorylase activity of the sulfate transporter. As such, the mutation may be considered to be a partial or loss-of-function mutation. Such mutations may be referred to herein as immunel (imm1) or low phytic acidl (Ipa1).

[0144] The method may not comprise introducing a mutation into any other gene. In other words, only the SULTR3;3 and / or SULTR3;4 gene and / or promoter is mutated. In one embodiment, both SULTR3;3 and SULTR3;4 genes are mutated.

[0145] In a preferred embodiment, the mutation that is introduced into the endogenous SULTR3;3 and / or SULTR3;4 gene or promoter thereof to silence, reduce, or inhibit the biological activity, function and / or expression levels of the SULTR3;3 and / or SULTR3;4 gene or protein can be selected from the following mutation types.

[0146] 1. a "missense mutation", which is a change in the nucleic acid sequence that results in the substitution of one amino acid for another amino acid;

[0147] 2. a "nonsense mutation" or " STOP codon mutation", which is a change in the nucleic acid sequence that results in the introduction of a premature STOP codon and, thus, the termination of translation (resulting in a truncated protein); in plants, the translation stop codons may be selected from " TGA" (UGA in RNA), " TAA" (UAA in RNA) and " TAG" (UAG in RNA); thus any nucleotide substitution, insertion, deletion which results in one of these codons to be in the mature mRNA being translated (in the reading frame) will terminate translation.M& C PC932446GB

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[0149] 3. an "insertion mutation" of one or more nucleotides or one or more amino acids, due to one or more codons having been added in the coding sequence of the nucleic acid;

[0150] 4. a "deletion mutation" of one or more nucleotides or of one or more amino acids, due to one or more codons having been deleted in the coding sequence of the nucleic acid;

[0151] 5. a "frameshift mutation", resulting in the nucleic acid sequence being translated in a different frame downstream of the mutation. A frameshift mutation can have various causes, such as the insertion, deletion or duplication of one or more nucleotides.

[0152] 6. a “splice site” mutation, which is a mutation that results in the insertion, deletion or substitution of a nucleotide at the site of splicing.

[0153] As used herein, an “insertion”, “deletion” or “substitution” may refer to the insertion, deletion or substitution of at least one, two, three, four, five, six, seven, eight, nine or ten nucleotides. In one specific embodiment, said mutation may comprise the substitution of at least one of the following positions, or corresponding positions in homologous sequences

[0154] S to F at position 371 of SEQ ID NO: 1 and / or C to T at position 1112 of SEQ ID NO: 73; and / or

[0155] G to E at position 316 of SEQ ID NO: 1 and / or G to A at position 947 of SEQ ID NO: 73; and / or

[0156] E to K at position 347 of SEQ ID NO: 1 and or G to A at position 1039 of SEQ ID NO: 73; and / or

[0157] Q to * at position 502 of SEQ ID NO: 1 and / or C to T at position 1504 of SEQ ID NO: 73; and / or

[0158] - A to V at position 381 of SEQ ID NO: 1 and / or C to T at position 1142 of SEQ ID NO: 73; and / or

[0159] L to F at position 413 of SEQ ID NO: 1 and or C to T at position 1237 SEQ ID NO: 73; and / or

[0160] P to L at position 478 of SEQ ID NO: 1 and / or C to T at position 1433 of SEQ ID NO: 73; and / or

[0161] P to S at position 587 of SEQ ID NO: 1 and / or C to T at position 1759 SEQ ID NO: 73; and / orM& C PC932446GB

[0162] 22

[0163] Q to * at position 230 of SEQ ID NO: 1 and / or C to T at position 688 SEQ ID NO: 73; and / or

[0164] G to R at position 327 of SEQ ID NO: 1 and / or G to A at position 979 of SEQ ID NO: 73.

[0165] In one embodiment, “insertion”, “deletion” or “substitution” may refer to the insertion, deletion, or substitution of at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% of a gene.

[0166] In another aspect of the invention there is provided a genetically altered plant, part thereof or plant cell characterised in that the expression of SULTR3;3 and SULTR3;4 is reduced or abolished or the plant does not express a functional or fully functional SULTR3;3 and SULTR3;4 protein.

[0167] In one embodiment, the plant is a reduction (knock down) or loss of function (knock out) mutant wherein the function of the SULTR3;3 and SULTR3;4 nucleic acid sequence is reduced or lost compared to a wild type or control plant. Preferably, the plant is a knockout and not a knock down, meaning that SULTR3;3 and SULTR3;4 expression is abolished or significantly abolished (e.g. by at least 80%, 90% or 95% or more) or that the plant expresses a SULTR3;3 and SULTR3;4 protein with no detectable function and / or activity (e.g. transport function). To this end, a mutation is introduced into the SULTR3;3 and / or SULTR3;4 gene sequence or the corresponding promoter sequence which disrupts the transcription of the gene or the function of the protein. The mutation may be any mutation that reduces or abolishes the transport activity of SULTR3;3 and / or SULTR3;4. For example, at least one of phosphorous, phosphate and / or sulfate transport.

[0168] In another aspect of the invention there is provided a genetically altered tomato plant, part thereof or plant cell characterised in that the expression of SULTR3;3 is reduced or abolished or the plant does not express a functional or fully functional SULTR3;3.

[0169] In one embodiment, the plant is a reduction (knock down) or loss of function (knock out) mutant wherein the function of the SULTR3;3 nucleic acid sequence is reduced or lost compared to a wild type or control plant. Preferably, the plant is a knock-out and not a knock down, meaning that SULTR3;3 expression is abolished or significantly abolishedM& C PC932446GB

[0170] 23

[0171] (e.g. by at least 80%, 90% or 95% or more) or that the plant expresses a SULTR3;3 and protein with no detectable function and / or activity (e.g. transport function). To this end, a mutation is introduced into the SULTR3;3 gene sequence or the corresponding promoter sequence which disrupts the transcription of the gene or the function of the protein. The mutation may be any mutation that reduces or abolishes the transport activity of SULTR3;3. For example, at least one of phosphorous, phosphate and / or sulfate transport.

[0172] In one embodiment, and as described above, the plant may be characterised by an increase in disease resistance, and in particular, an increase in fungal disease resistance. The plant may additionally be characterised by a decrease in phytic acid levels, as described above. The plant may further have no growth or no significant growth penalties - that is, no difference in growth or yield compared to a plant that has not been genetically altered.

[0173] In one embodiment, the genetically altered plant is further characterised by an increase in total seed weight per plant compared to a wild-type plant. As shown in Figure 7B, the Arabidopsis imm1 a mutant shows increased seed weight per plant compared to the wildtype plant and also the mlo2-5 mutant. Accordingly, there is provided a method of increasing resistance to disease, preferably fungal disease, and further increasing seed weight per plant compared to a control. There is also provided a method of increasing resistance to disease, preferably fungal disease, and decreasing phytic acid levels in the plant, and increasing seed weight per plant compared to a control.

[0174] By increasing seed weight is meant an increase in total seed weight per plant compared to a wild-type or control plant. By increase, is meant an increase of at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%.

[0175] Preferably said mutation is introduced into a least one plant cell and a plant regenerated from the at least one mutated plant cell.

[0176] The mutation may be introduced into the coding region of a SULTR3;3 and / or SULTR3;4 gene. Alternatively, the mutation may be in an intronic sequence, a splice site, the 5’IITR or 3’IITR or a regulatory site of any such sites. In a further alternative, at least oneM& C PC932446GB

[0177] 24

[0178] mutation or structural alteration may be introduced into the SULTR3;3 and / or SULTR3;4 promoter such that the SULTR3;3 and / or SULTR3;4 gene is either not expressed (i.e. expression is abolished) or expression is reduced, as defined herein. In an alternative embodiment, at least one mutation may be introduced into the SULTR3;3 and / or SULTR3;4 gene such that the altered gene does not express a full-length (i.e. expresses a truncated) SULTR3;3 and / or SULTR3;4 protein or does not express a fully functional SULTR3;3 and / or SULTR3;4 protein. In this manner, the (sulfate, phosphate and / or phosphorous transport) activity of the SULTR3;3 and / or SULTR3;4 polypeptide can be considered reduced or abolished as described herein. In any case, the mutation may result in the expression of SULTR3;3 and / or SULTR3;4 with no, significantly reduced or altered biological activity in vivo.

[0179] By “at least one mutation” is meant that where the SULTR3;3 and / or SULTR3;4 gene is present as more than one copy or homoeologue (with the same or slightly different sequence) there is at least one mutation in at least one gene. Preferably, all genes or all homeologs are mutated. For example, to achieve a reduction or abolishment in SULTR3;3 and / or SULTR3;4 gene in wheat, at least one mutation must be present in each of the hexapioid chromosomes.

[0180] In one embodiment, the nucleic acid sequence of the SULTR3;3 gene comprises or consists of a nucleic acid sequence selected from Table 1.

[0181] In one embodiment, the nucleic acid sequence of the SULTR3;3 gene comprises or consists of a nucleic acid sequence that encodes a polypeptide as defined in one of SEQ ID NO: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, or 54-56 respectively or a functional variant or homolog thereof.

[0182] In one embodiment, the amino acid sequence of SULTR3;3 comprises or consists of a sequence as defined in one of SEQ ID NO: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52 or 54-56 respectively or a functional variant or homolog thereof.

[0183] In one embodiment, the nucleic acid sequence of the SULTR3;4 gene comprises or consists of a nucleic acid sequence selected from Table 1.M& C PC932446GB

[0184] 25

[0185] In one embodiment, the nucleic acid sequence of the SULTR3;4 gene comprises or consists of a nucleic acid sequence that encodes a polypeptide as defined in one of SEQ ID NO: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 respectively or a functional variant or homolog thereof.

[0186] In one embodiment, the amino acid sequence of the SULTR3;4 comprises or consists of a sequence as defined in one of SEQ ID NO: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, or 56 or a functional variant or homolog thereof.

[0187] As used throughout, by “SULTR3;3 promoter” is meant a region extending at least or approx. 10 kbp, more preferably about 6 kbp upstream of the ATG codon of the SULTR3;3 ORF. In one embodiment, the sequence of the SULTR3;3 promoter comprises or consists of a nucleic acid sequence as defined by a sequence in Table 1. Where the plant is barely, the SULTR3;3 promoter may comprise or consist of residues 1 to 1000 of SEQ ID NO: 71.

[0188] In one embodiment, the SULTR3;3 promoter may also include 5’ UTR sequences.

[0189] As used throughout, by “SULTR3;4 promoter” is meant a region extending at least or approx.10 kbp, more preferably about 7 kbp upstream of the ATG codon of the SULTR3;4 ORF. In one embodiment, the sequence of the SULTR3;4 promoter comprises or consists of a nucleic acid sequence as defined by a sequence in Table 1. In one embodiment, the SULTR3;4 promoter may also include 5’ UTR sequences. Where the plant is barley, the SULTR3;4 promoter may comprise or consist of residues 1 to 1283 of SEQ ID NO: 72.

[0190] In the above embodiments an ‘endogenous’ nucleic acid may refer to the native or natural sequence in the plant genome. Also included in the scope of this invention are functional variants (as defined herein) and homologs of the above identified sequences. Examples of homologs are shown in SEQ ID NOs: 1-56.

[0191] The homolog of SULTR3;3 may be from wheat, maize, rice, barley, oat, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, soybean, sugarbeet, grape, sunflower, sugar cane and encodes a polypeptide selected from SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52 and 54-56; or the homolog comprises or consists of aM& C PC932446GB

[0192] 26

[0193] nucleic acid sequence selected from a sequence in Table 1. In another embodiment, the SULTR3;3 promoter homolog comprises or consists of a nucleic acid sequence as defined in Table 1.

[0194] The homolog of SULTR3;4 may be from wheat, maize, rice, barley, oat, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, soybean, sugarbeet, grape, sunflower, sugar cane and encodes a polypeptide selected from SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55 and 56; or the homolog comprises or consists of a nucleic acid sequence selected from a sequence in Table 1. In another embodiment, the SULTR3;4 promoter homolog comprises or consists of a nucleic acid sequence as defined in Table 1.

[0195] In one embodiment, the plant is barley and the SULTR3;3 encodes a polypeptide selected from SEQ ID NOs: 3 or 4; or the homolog comprises or consists of a nucleic acid sequence selected from SEQ ID NO: 71. In another embodiment, the SULTR3;3 promoter homolog comprises or consists of a nucleic acid sequence as defined in residues 1 to 1000 of SEQ ID NO: 71.

[0196] In one embodiment, the plant is barley and the SULTR3;4 encodes a polypeptide selected from SEQ ID NOs: 5; or the homolog comprises or consists of a nucleic acid sequence selected from SEQ ID NO: 72. In another embodiment, the SULTR3;4 promoter homolog comprises or consists of a nucleic acid sequence as defined in residues 1 to 1283 of SEQ ID NO: 72.

[0197] In one embodiment, the plant is tomato and the SULTR3;3 encodes a polypeptide as defined in SEQ ID NO: 39. As shown in Figure 17, only a mutation in the SULTR3;3 gene is required to confer powdery mildew resistance.

[0198] Accordingly, for any of the method aspects of the invention, where the plant is tomato, the method comprises reducing or abolishing the expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3 member. Preferably, the SLILTR 3;3 comprises an amino acid sequence defined in SEQ ID NO: 39 or a functional variant or homolog thereof.M& C PC932446GB

[0199] 27

[0200] In one embodiment, the method comprises introducing at least one mutation into at least one gene encoding a SULTR3;3 protein and / or introducing at least one mutation into a SULTR3;3 promoter, wherein said mutation results in reduced expression and / or activity of SULTR3;3 compared to a wild-type or control plant.

[0201] The term “functional variant” (or “variant”) as used herein with reference to any of the sequences described herein refers to a variant sequence or part of the sequence which retains the biological function of the full non-variant sequence. Accordingly, in the context of SULTR3;3 and / or SULTR3;4, a functional variant retains the same or similar transporter function of a given SULTR3;3 and / or SULT3;4. In one embodiment, the functional variant may transport sulfate, a phosphate and / or phosphorous transporter. As used herein, a homolog may also be referred to as functional. That is, the homolog is a sulfate, phosphate and / or phosphorous transporter. Levels of phosphate, phosphorous or sulfate transport can be measured using any known technique in the art, for example, using by placing a plant sample or a plant cell in medium containing a known amount of radioactive sulfate, phosphate and / or phosphorous and using mass spectrometry to quantify the sulfate, phosphate and / or phosphorous levels before and after a time period, to provide a measurement of ion uptake and transporter activity. Alternatively, a FRET-based sensor can be used to assess transporter activity by providing a real-time readout of how much substrate accumulates inside the cell as the transporter moves it across the membrane. The sensor consists of two fluorescent proteins connected by a substrate-binding domain; when the substrate enters the cell and binds the sensor, the resulting conformational change alters the FRET ratio between the donor and acceptor fluorophores. By expressing the transporter in the plasma membrane and the FRET sensor in the cytosol (or another compartment), then adding the transporter’s substrate externally, it is possible to track changes in the FRET signal over time. An increase in the FRET ratio reflects rising intracellular substrate levels, indicating active transport, whereas no change suggests that the transporter is inactive or absent. By comparing the transport activity and kinetics of a variant transporter to a control transporter, it is possible to determine if a variant retains the same function as the non-variant transporter. The gold-standard assay for analysing the function of a transporter involves the insertion of said transporter into xenopus oocytes (exemplar protocol described in Ding et al. (2020)) and then using a FRET-based sensor or radioactive ion uptake assay as described above.M& C PC932446GB

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[0203] Fang et al. (2025) have determined that a serine at position 170 of SEQ ID NO: 7 is a key determinant of substrate specificity and that this is present in SLILTR 3;3 and SLILTR 3;4 transporters. As described herein, the residue is not essential for transport (a mutation at this position only affected Pi transport, but not sulfate transport) but it is a key determinant for substrate selectivity. Therefore, it is preferable that a variant would have a threonine or serine at this position because it is indicative of having Pi and sulfate transport function.

[0204] In one embodiment, the variant comprises a serine or threonine at a position corresponding to any one of: position 170 of SEQ ID NO: 7, position 124 of SEQ ID NO: 1 or position 147 of SEQ ID NO: 2. Preferably, the variant comprises a serine at the corresponding position. In one embodiment, the variant comprises a SIASL or TIASL motif in the amino acid sequence, preferably an SIASL motif.

[0205] The functional variant may also comprise variants of the SULTR3;3 and / or SULTR3;4 gene, which have sequence alterations that do not affect function, for example in nonconserved 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 (e.g. has phosphorylase activity). Alterations in a nucleic acid sequence which result in the production of a different amino acid at a given site that do 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.

[0206] As used herein, the term “functional variant” also encompasses a functional fragment. A “functional fragment” refers to a functionally active series of consecutive amino acidsM& C PC932446GB

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[0208] from a longer polypeptide or protein. For example, a functional fragment may still function as a sulfate, phosphate and / or phosphorous transporter.

[0209] In one embodiment, a functional variant has at least 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to the non-variant nucleic acid or amino acid sequence.

[0210] The term homolog, as used herein, also designates a SULTR3;3 and / or SULTR3;4 promoter or SULTR3;3 and / or SULTR3;4 gene orthologue from other plant species. A homolog may have, in increasing order of preference, at least 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to the amino acid represented by any of SEQ ID NO: 1-57, or to the nucleic acid sequences listed in Table 1, SEQ ID NO: 71-74. A SULTR3;3 and / or SULTR3;4 promoter orthologue may have, in increasing order of preference, at least 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to the nucleic acid sequences as shown in SEQ ID Nos: 71-74 or a sequence listed in Table 1.

[0211] 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 or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when comparedM& C PC932446GB

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[0213] 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 acids 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. Non-limiting examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms. The overall sequence identity of a variant can be determined using any number of sequence alignment programs known in the art. As an example, Emboss Stretcher from the EMBL-EBI may be used: https: / / www.ebi.ac.uk / Tools / psa / emboss stretcher / (using default parameters: pair output format, Matrix = BLOSUM62, Gap open = 1, Gap extend = 1 for proteins; pair output format, Matrix = DNAfull, Gap open = 16, Gap extend = 4 for nucleotides).

[0214] The skilled person would understand that suitable homologues and the homologous positions in these sequences can be identified by sequence comparisons (e.g. BLAST, alignments) and identifications of conserved domains. Phylogenetic tree analysis using nucleotide or amino acid sequences can be used to establish orthology to SULTR3;3 and / or SULTR3;4. There are predictors in the art that can be used to identify such sequences. The function of the homolog can be identified as described herein and a skilled person would thus be able to confirm the function, for example by measuring levels of sulfate, phosphate and / or phosphorous transport.M& C PC932446GB

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[0216] 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).

[0217] 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.

[0218] 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 24 hours, usually about 4 to 12. StringentM& C PC932446GB

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[0220] conditions may also be achieved with the addition of destabilizing agents such as formamide.

[0221] In a further embodiment, a variant as used herein can comprise a nucleic acid sequence encoding a SULTR3;3 and / or SULTR3;4 polypeptide as defined herein that is capable of hybridising under stringent conditions as defined herein to a nucleic acid sequence as defined herein, such as a sequence listed in Table 1 or SEQ ID NO: 71-74, or a nucleic acid sequence that encodes a polypeptide defined in SEQ ID NO: 1-57.

[0222] In one embodiment, the method comprises reducing or abolishing the expression of at least one nucleic acid encoding a SULTR3;3 and / or SULTR3;4 polypeptide or reducing or abolishing the activity of an SULTR3;3 and / or SULTR3;4 polypeptide, as described herein, wherein the method comprises introducing at least one mutation into at least one SULTR3;3 and / or SULTR3;4 gene and / or promoter, wherein the SULTR3;3 gene comprises or consists of

[0223] a. a nucleic acid sequence encoding a polypeptide as defined in one of SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56 or

[0224] b. a nucleic acid sequence as defined in one of Table 1 or SEQ ID Nos: 71 or 73 or c. a nucleic acid sequence with at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to either (a) or (b); or d. a nucleic acid sequence encoding a SULTR3;3 polypeptide as defined herein that is capable of hybridising under stringent conditions as defined herein to the nucleic acid sequence of any of (a) to (c);

[0225] and wherein the SULTR3;3 promoter comprises or consists of

[0226] e. a nucleic acid sequence as defined in Table 1 or SEQ ID Nos: 71 or 73; or f. a nucleic acid sequence with at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to (e); or

[0227] g. a nucleic acid sequence for a SULTR3;3 promoter as defined herein that is capable of hybridising under stringent conditions as defined herein to the nucleic acid sequence of any (e) to (f);

[0228] and wherein the SULTR3;4 gene comprises or consists of

[0229] h. a nucleic acid sequence encoding a polypeptide as defined in one of SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55 or 56; orM& C PC932446GB

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[0231] i. a nucleic acid sequence as defined in Table 1 or SEQ ID NO: 72 or 74; or j. a nucleic acid sequence with at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to either (h) or (i); or k. a nucleic acid sequence encoding a SULTR3;4 polypeptide as defined herein that is capable of hybridising under stringent conditions as defined herein to the nucleic acid sequence of any of (h) to (j);

[0232] and wherein the SULTR3;4 promoter comprises or consists of

[0233] l. a nucleic acid sequence as defined in Table 1 or SEQ ID NO: 72 or 74; m. a nucleic acid sequence with at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% overall sequence identity to (I); or

[0234] n. a nucleic acid sequence for a SULTR3;4 promoter as defined herein that is capable of hybridising under stringent conditions as defined herein to the nucleic acid sequence of any of (I) to (m).

[0235] In one embodiment, a mutation is introduced into a SULTR3;3 and / or a SULTR3;4 gene, wherein the first SULTR3;3 gene encodes a polypeptide selected from SEQ ID NO: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56 (preferably the SULTR3;3 gene comprises or consists of a nucleic acid sequence selected from SEQ ID NO: 71 or 73 or a sequence in Table 1) and / or wherein the SULTR3;4 gene encodes a polypeptide selected from SEQ ID NO: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 (preferably the SULTR3;4 gene comprises or consists of a nucleic acid sequence selected, from SEQ ID NO: 72 or 74 or a sequence in Table 1).

[0236] In a preferred embodiment, the mutation that is introduced into the endogenous SULTR3;3 and / or SULTR3;4 gene or promoter thereof to alter the biological activity and / or expression levels of the SULTR3;3 and / or SULTR3;4 gene or protein can be selected from the following mutation types:

[0237] 1. a "missense mutation", which is a change in the nucleic acid sequence that results in the substitution of one amino acid for another amino acid;

[0238] 2. a "nonsense mutation" or " STOP codon mutation", which is a change in the nucleic acid sequence that results in the introduction of a premature STOP codon and, thus, the termination of translation (resulting in a truncated protein); inM& C PC932446GB

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[0240] plants, the translation stop codons may be selected from " TGA" (UGA in RNA), " TAA" (UAA in RNA) and " TAG" (UAG in RNA); thus any nucleotide substitution, insertion, deletion which results in one of these codons to be in the mature mRNA being translated (in the reading frame) will terminate translation.

[0241] 3. an "insertion mutation" of one or more nucleotides or one or more amino acids, due to one or more codons having been added in the coding sequence of the nucleic acid;

[0242] 4. a "deletion mutation" of one or more nucleotides or of one or more amino acids, due to one or more codons having been deleted in the coding sequence of the nucleic acid;

[0243] 5. a "frameshift mutation", resulting in the nucleic acid sequence being translated in a different frame downstream of the mutation. A frameshift mutation can have various causes, such as the insertion, deletion or duplication of one or more nucleotides.

[0244] 6. a “splice site” mutation, which is a mutation that results in the insertion, deletion or substitution of a nucleotide at the site of splicing (i.e. either a splice acceptor or splice donor mutation)

[0245] where any one or more of the above mutations leads to a loss or partial loss of function in SULTR3;3 and / or SULTR3;4 (e.g. a complete or partial loss of sulfate, phosphate and / or phosphorous transport activity). The degree to which function is lost can vary considerably. A complete or total loss of function is when all of the wild-type function of the allele is lost, i.e. no function remains. A partial loss of function refers to when some of the function remains, but not at the same level as the wild-type allele, the function that remains is less than that of wild-type.

[0246] In particular, we have found at a mutation at a position close to the border between sequences encoding a transmembrane domain and sequences encoding cytosolic or extracellular loops are very important for the function of a SULTR3;3 and SULTR3;4.

[0247] By “partial loss” is meant reducing the function to a level lower than that in a wild-type plant. The term “partial loss” means a decrease in the function by up to 5% 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% when compared to the level of function in a control plant.M& C PC932446GB

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[0249] In general, the skilled person will understand that at least one mutation as defined above and which leads to the insertion, deletion or substitution of at least one nucleic acid or amino acid compared to the wild-type SULTR3;3 and / or SULTR3;4 promoter or SULTR3;3 and / or SULTR3;4 nucleic acid or protein sequence can affect the biological activity of the SULTR3;3 and / or SULTR3;4 protein.

[0250] In one embodiment, the mutation is introduced using mutagenesis or targeted genome editing. That is, in one embodiment, the invention relates to a method and plant that has been generated by genetic engineering methods as described above, and does not encompass naturally occurring varieties.

[0251] 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. To achieve effective genome editing via introduction of site-specific DNA DSBs, four major classes of customisable DNA binding proteins can be used: meganucleases derived from microbial mobile genetic elements, ZF nucleases based on eukaryotic transcription factors, 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 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.

[0252] Preferably the 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 and references cited herein.

[0253] Alternatively, more conventional mutagenesis methods can be used to introduce at least one mutation into a SULTR3;3 and / or SULTR3;4 gene or SULTR3;3 and / or SULTR34 promoter sequence. These methods include both physical and chemical mutagenesis. A skilled person will know further approaches can be used to generate such mutants, andM& C PC932446GB

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[0255] methods for mutagenesis and polynucleotide alterations are well known in the art. See, for example, Kunkel (1985) Proc. Natl. Acad. Sci. USA 82:488-492; Kunkel et al. (1987) Methods in Enzymol. 154:367-382; U. S. Patent No. 4,873,192; Walker and Gaastra, eds. (1983) Techniques in Molecular Biology (MacMillan Publishing Company, New York) and the references cited therein.

[0256] In one embodiment, insertional mutagenesis is used, for example using T-DNA mutagenesis (which inserts pieces of the T-DNA from the Agrobacterium tumefaciens Ti-Plasmid into DNA causing loss of gene function mutations); site-directed nucleases (SDNs) or transposons as a mutagen. Insertional mutagenesis is an alternative means of disrupting gene function and is based on the insertion of foreign DNA into the gene of interest (see Krysan et al, The Plant Cell, Vol. 11, 2283-2290, December 1999). Accordingly, in one embodiment, T-DNA is used as an insertional mutagen to disrupt a SULTR3;3 and / or SULTR3;4 gene or SULTR3;3 and / or SULTR3;4 promoter expression. T-DNA not only disrupts the expression of the gene into which it is inserted, but also acts as a marker for subsequent identification of the mutation. Since the sequence of the inserted element is known, the gene in which the insertion has occurred can be recovered, using various cloning or PCR-based strategies. The insertion of a piece of T-DNA in the order of 5 to 25 kb in length generally produces a disruption of gene function. If a large enough population of T-DNA transformed lines is generated, there are reasonably good chances of finding a transgenic plant carrying a T-DNA insert within any gene of interest. Transformation of spores with T-DNA is achieved by an Agrobacterium-mediated method which involves exposing plant cells and tissues to a suspension of Agrobacterium cells.

[0257] 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 mutant 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 SULTR3;3 and / or SULTR3;4 nucleic acid sequence in a wild type plant and phenotypic analysis is also carried out.M& C PC932446GB

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[0259] In another embodiment, mutagenesis is physical mutagenesis, such as application of ultraviolet radiation, X-rays, gamma rays, fast or thermal neutrons or protons. The targeted population can then be screened to identify an SULTR3;3 and / or SULTR3;4 mutant with reduced expression or activity.

[0260] In another embodiment of the various aspects of the invention, the method comprises mutagenizing a plant population with a mutagen. The mutagen may be a fast neutron irradiation or a chemical mutagen, for example selected from the following non-limiting list: ethyl methanesulfonate (EMS), methylmethane sulfonate (MMS), N-ethyl-N-nitrosurea (ENU), triethylmelamine (TEM), N-methyl-N-nitrosourea (MNU), procarbazine, chlorambucil, cyclophosphamide, diethyl sulfate, acrylamide monomer, melphalan, nitrogen mustard, vincristine, dimethylnitosamine, N-methyl-N'-nitro-Nitrosoguanidine (MNNG), nitrosoguanidine, 2-aminopurine, 7,12 dimethyl-benz(a)anthracene (DMBA), ethylene oxide, hexamethylphosphoramide, bisulfan, diepoxyalkanes (diepoxyoctane (DEO), diepoxybutane (BEB), and the like), 2-methoxy-6-chloro-9 [3-(ethyl-2-chloroethyl)aminopropylamino]acridine dihydrochloride (ICR-170) or formaldehyde. Again, the targeted population can then be screened to identify a SULTR3;3 and / or SULTR3;4 gene or promoter mutant.

[0261] In another embodiment, the method used to create and analyse mutations is targeting induced local lesions in genomes (TILLING), reviewed in Henikoff et al, 2004.

[0262] Subsequently, rapid high-throughput screening procedures allow the analysis of amplification products for identifying a mutation conferring the reduction or inactivation of the expression of the SULTR3;3 and / or SULTR3;4 gene as compared to a corresponding non-mutagenised wild type plant. Once a mutation is identified in a gene of interest, the seeds of the M2 plant carrying that mutation are grown into adult M3 plants and screened for the phenotypic characteristics associated with the target gene SULTR3;3 and / or SULTR3;4. Loss of and reduced function mutants with an increased disease resistance compared to a control can thus be identified.

[0263] In an alternative embodiment, the expression of the SULTR3;3 and / or SULTR3;4 gene may be reduced at either the level of transcription or translation. For example, expression of a SULTR3;3 and / or SULTR3;4 gene sequence, as defined herein, can be reduced or silenced using a number of gene silencing methods known to the skilled person, suchM& C PC932446GB

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[0265] as, but not limited to, the use of small interfering nucleic acids (siNA) against SULTR3;3 and / or SULTR3;4. “Gene silencing" is a term generally used to refer to suppression of expression of a gene via sequence-specific interactions that are mediated by RNA molecules. The degree of reduction may be so as to totally abolish production of the encoded gene product, but more usually the abolition of expression is partial, with some degree of expression remaining. The term should not therefore be taken to require complete "silencing" of expression.

[0266] In one embodiment, the siRNA may include, short interfering RNA (siRNA), doublestranded RNA (dsRNA), micro-RNA (miRNA), antagomirs and short hairpin RNA (shRNA) capable of mediating RNA interference. The inhibition of expression and / or activity can be measured by determining the presence and / or amount of SULTR3;3 and / or SULTR3;4 transcript using techniques well known to the skilled person (such as Northern Blotting, RT-PCR and so on).

[0267] Thus, according to the various aspects of the invention a plant may be transformed to introduce a RNAi, shRNA, snRNA, dsRNA, siRNA, miRNA, ta-siRNA, amiRNA or cosuppression molecule that has been designed to target the expression of an SULTR3;3 and / or 3;4 nucleic acid sequence and selectively decreases or inhibit the expression of the gene or stability of its transcript. Preferably, the RNAi, snRNA, dsRNA, shRNA siRNA, miRNA, amiRNA, ta-siRNA or cosuppression molecule used according to the various aspects of the invention comprises a fragment of at least 17 nt, preferably 22 to 26 nt and can be designed on the basis of the information shown in any of SEQ ID NOs: 1-57 and 58-70. Guidelines for designing effective siRNAs are known to the skilled person.

[0268] Plants obtained or obtainable by such method which carry a mutation in the endogenous SULTR3;3 and / or SULTR3;4 gene or promoter locus are also within the scope of the invention, where preferably the mutation is a loss or partial loss of function mutation as described above.

[0269] Accordingly, the genetically altered plant or plant cell may alternatively comprise a nucleic acid construct expressing an RNAi molecule,, targeting the SULTR3;3 and / or SULTR3;4 gene as described herein. In one embodiment, said construct is stably incorporated into the plant genome.M& C PC932446GB

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[0271] In another aspect of the invention there is provided a method for producing a genetically altered plant as described herein. In one embodiment, the method comprises introducing at least one mutation into the SULTR3;3 and / or SULTR3;4 gene and / or SULTR3;3 and / or SULTR3;4 promoter of preferably at least one plant cell using any mutagenesis technique described herein. Preferably said method further comprising regenerating a plant from the mutated plant cell.

[0272] The method may further comprise selecting one or more mutated plants, preferably for further propagation. Preferably said selected plants comprise at least one mutation in the SULTR3;3 and / or SULTR3;4 gene and / or promoter sequence. Preferably said plants are characterised by reduced or abolished SULTR3;3 and / or SULTR3;4 expression and / or a reduced or abolished levels of SULTR3;3 and / or SULTR3;4 polypeptide activity. Expression and / or activity levels (e.g. sulfate, phosphate and / or phosphorous transport level) of SULTR3;3 and / or SULTR3;4 can be measured by any standard technique known to the skilled person, including those described herein.

[0273] The selected 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. The generated transformed organisms may take a variety of forms. For example, they may be chimeras of transformed cells and non-transformed cells; clonal transformants (e.g., all cells transformed to contain the expression cassette); grafts of transformed and untransformed tissues (e.g., in plants, a transformed rootstock grafted to an untransformed scion).

[0274] A genetically altered plant of the present invention may also be obtained by transference of any of the sequences of the invention by crossing, e.g., using pollen of the genetically altered plant described herein to pollinate a wild-type or control plant, or pollinating the gynoecia of plants described herein with other pollen that does not contain a mutation in at least one of the SULTR3;3 and SULTR3;4 gene or promoter sequence. The methods for obtaining the plant of the invention are not exclusively limited to those described in this paragraph; for example, genetic transformation of germ cells from the ear of wheat could be carried out as mentioned, but without having to regenerate a plant afterward.M& C PC932446GB

[0275] 40

[0276] In a further aspect of the invention there is provided a plant obtained or obtainable by the above described methods.

[0277] For the purposes of the invention, a “genetically altered plant” or “mutant plant” is a plant that has been genetically altered compared to the naturally occurring wild type (WT) plant. In one embodiment, a mutant plant is a plant that has been altered compared to the naturally occurring wild type (WT) plant using a mutagenesis method, such as any of the mutagenesis methods described herein. In one embodiment, the mutagenesis method is targeted genome modification or genome editing. In one embodiment, the plant genome has been altered compared to wild type sequences using a mutagenesis method. Such plants have an altered phenotype as described herein, such as an increased disease resistance, particularly fungal disease resistance. Therefore, in this example, increased disease resistance is conferred by the presence of an altered plant genome, for example, a mutated endogenous SULTR3;3 and / or SULTR3;4 gene or SULTR3;3 and / or SULTR3;4 promoter sequence. In one embodiment, the endogenous promoter or gene sequence is specifically targeted using targeted genome modification and the presence of a mutated gene or promoter sequence is not conferred by the presence of transgenes expressed in the plant. In other words, the genetically altered plant can be described as transgene-free.

[0278] A plant according to the various aspects of the invention, including the transgenic plants, methods and uses described herein may be a monocot or a dicot plant. Preferably, the plant is a crop plant or a biofuel 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 vegetable. In a preferred embodiment, the plant is a cereal. In one embodiment the plant is not Arabidopsis.

[0279] Preferably the plant is selected from wheat, maize, rice, barley, rye, oat, millet, corn, sorghum, sugar cane, ryegrass, oil seed rape, brassica, potato, okra, carrot, parsnip, turnip, Lactuca, Cucurbits (such as pumpkin, cucumber, tomato, watermelon, melon, zucchini, squash, pumpkin, gourd etc.), tomato, pepper, aubergine (eggplant), legumes (such as, pea, faba bean, common bean, soybean, chickpes etc.) cotton, sunflower, sugarbeet, grape, sunflower, oil palm, apple, peach, orange, lemon, blueberry and quinoa.M& C PC932446GB

[0280] 41

[0281] In one embodiment, the crop plant is selected from wheat, maize, rice, barley, rye, oat millet, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, carrot, parsnip, turnip, zucchini, pumpkin, cucumber, tomato, watermelon, melon, zucchini, squash, pumpkin, gourd, pea, faba bean, common bean, soybean, chickpea, sugarbeet, grape, sunflower, rye, sugar cane and ryegrass.

[0282] Preferably the plant is cereal crop. In one embodiment, the cereal crop is selected from wheat, maize, rice, barley, rye, oat, millet, corn, sorghum, sugar cane, ryegrass, oil seed rape, brassica, soybean. Most preferably, the cereal crop is barley.

[0283] In a preferred embodiment, the plant is sugarbeet.

[0284] Trees are also susceptible to powdery mildew attacks, with the most commonly affected included oak, maple, dogwood, magnolia, catalpa and apple. Accordingly, in one embodiment the plant is a tree. In one embodiment, the tree is selected from oak, maple, dogwood, magnolia, catalpa and apple.

[0285] The term "plant" as used herein encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, fruit, shoots, stems, leaves, roots (including tubers), flowers, tissues and organs, wherein each of the aforementioned comprise the nucleic acid construct as described herein. The term "plant" also encompasses plant cells, suspension cultures, callus tissue, embryos, meristematic regions, gametophytes, sporophytes, pollen and microspores, again wherein each of the aforementioned comprises the nucleic acid construct as described herein.

[0286] The invention also extends to harvestable parts of a plant of the invention as described herein, but not limited to seeds, leaves, fruits, flowers, stems, roots, rhizomes, tubers and bulbs. The aspects of the invention also extend to products derived, preferably directly derived, from a harvestable part of such a plant, such as dry pellets or powders, oil, fat and fatty acids, starch or proteins. Another product that may derived from the harvestable parts of the plant of the invention is biodiesel. The invention also relates to food products and food supplements comprising the plant of the invention or parts thereof. In one embodiment, the food products may be animal feed. In another aspect of the invention, there is provided a product derived from a plant as described herein or from a part thereof.M& C PC932446GB

[0287] 42

[0288] The terms “seed” and “grain” as used herein can be used interchangeably. The terms "increase", "improve" or "enhance" as used herein are also interchangeable.

[0289] In a most preferred embodiment, the plant part or harvestable product is a seed or grain. Therefore, in a further aspect of the invention, there is provided a seed or grain produced from a genetically altered plant as described herein. Preferably the seed or grain comprises reduced expression or activity of a SULTR3;3 and / or SULTR3;4 transporter. For example, the seed or grain comprise at least one mutation, preferably a loss of function mutation, in a SULTR3;3 and / or SULTR3;4 gene or promoter sequence.

[0290] In an alternative embodiment, the plant part is pollen, a propagule or progeny of the genetically altered plant described herein. Accordingly, in a further aspect of the invention there is provided pollen, a propagule or progeny of the genetically altered plant as described herein.

[0291] A control plant as used herein according to all of the aspects of the invention is a plant which has not been modified according to the methods of the invention. Accordingly, in one embodiment, the control plant does not have altered expression of a SULTR3;3 and / or SULTR3;4 nucleic acid and / or altered activity of a SULTR3;3 and / or SULTR3;4 polypeptide, as described herein. In an alternative embodiment, the plant been genetically modified, as described above. In one embodiment, the control plant is a wild type plant. The control plant is typically of the same plant species, preferably having the same genetic background as the modified plant.

[0292] Genome editing constructs for use with the methods for targeted genome modification described herein

[0293] We have designed a CRISPR / Cas9 guide RNA construct for simultaneous mutation of IMM1A and IMM1B in barley. The construct comprised oligos, IMM1ABC20F: 5’-ACTTGGAGGCTGGCAATGGTGAGGCG-3’ and IMM1ABC20R: 5’- AAAACGCCTCACCATTGCCAGCCTCC-3’, which were annealed and GATEWAY™ cloned into the T-DNA destination pANIC6A vector while recombining with the entry vectors, pJG85 and pJG80, for combining sequences for sgRNA and Cas9 expression.M& C PC932446GB

[0294] 43

[0295] Accordingly, in another aspect of the invention there is provided a nucleic acid construct wherein the nucleic acid construct comprises a nucleic acid sequence that encodes at least one DNA-binding domain. In one embodiment, the DNA-binding domain can bind to a sequence in the SULTR3;3 and / or SULTR3;4 gene and / or promoter. In one embodiment, the nucleic acid construct comprises one or more DNA-binding domains, such that the construct can bind to one or more, preferably at least two sequences in the SULTR3;3 and / or SULTR3;4 genes. Preferably said sequence is selected from one or both of SEQ ID NO: 83 and 84 and are target sequences in a SULTR3;3 and SULTR3;4 gene.

[0296] In a further embodiment, said construct further comprises a nucleic acid encoding at least one sequence specific nuclease (SSN) such as Fokl or a Cas protein.

[0297] In a further final aspect of the invention, there is provided a method of screening a population of plants and identifying and / or selecting a plant that will have increased disease resistance, and in particular, increased fugal disease resistance, as described herein, compared to a control or wild-type plant, the method comprising detecting at least one polymorphism or mutation in the SULTR3;3 and / or SULTR3;4 gene and / or promoter, wherein said mutation or polymorphism leads to an reduction in the level of expression and / or activity of the SULTR3;3 and / or SULTR3;4 protein compared to the level in a plant not carrying said mutation or polymorphism (e.g. a control or wild-type plant). Said mutation or polymorphism may comprise at least one insertion and / or at least one deletion and / or substitution.

[0298] Suitable tests for assessing the presence of a polymorphism would be well known to the skilled person, and include but are not limited to, Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length polymorphisms (AFLPs), Simple Sequence Repeats (SSRs-which are also referred to as Microsatellites), and Single Nucleotide Polymorphisms (SNPs). In one embodiment, Kompetitive Allele Specific PCR (KASP) genotyping is used.

[0299] In one embodiment, the method comprises

[0300] a) obtaining a nucleic acid sample from a plant andM& C PC932446GB

[0301] 44

[0302] b) carrying out nucleic acid amplification of one or more SULTR3;3 and / or SULTR3;4 gene and / or promoter alleles using one or more primer pairs.

[0303] In a further embodiment, the method may further comprise introgressing the chromosomal region comprising at least one of said SULTR3;3 and / or 3;4-expressing / activity polymorphisms into a second plant or plant germplasm to produce an introgressed plant or plant germplasm. Preferably the expression or activity of SULTR3;3 and / or SULTR3;4 in said second plant will be altered (compared to a control or wild-type plant), and more preferably said second plant will display increased disease resistance, as described above.

[0304] 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.

[0305] "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.

[0306] 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.

[0307] The foregoing application, and all documents and sequence accession numbers cited therein or during their prosecution ("appln cited documents") and all documents cited or referenced in the appln cited documents, and all documents cited or referenced herein ("herein cited documents"), and all documents cited or referenced in herein cited documents, together with any manufacturer's instructions, descriptions, productM& C PC932446GB

[0308] 45

[0309] specifications, and product sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated herein by reference, and may be employed in the practice of the invention. More specifically, all referenced documents are incorporated by reference to the same extent as if each individual document was specifically and individually indicated to be incorporated by reference.

[0310] The invention is now described in the following non-limiting examples.

[0311] EXAMPLE 1: Knock-out of the sulfate transporter, SULTR3;3, causes EDS1-independent resistance to powdery mildew.

[0312] In the present study, we performed a mutant screen for powdery mildew resistance in Arabidopsis. However, unlike previous screens, we made the mutant population in an eds1 mutant background in order to decrease the likelihood of identifying mutants that are resistant because of harmful activation of PTI and TNL-based ETI. This allowed us to identify an allelic series of homozygous immunela (immla) mutants with a significant level of resistance to Golovinomyces orontii (Go), a powdery mildew fungal pathogen of Arabidopsis. IMM1a was identified as SULTR3;3 encoding a sulfate transporter. While Arabidopsis immla plants have enhanced resistance to more fungal leaf pathogens, transient silencing of a close homolog in the mutant background caused enhanced resistance to powdery mildew in barley (Figure 14).

[0313] IMM1a (At1g23090) encodes the sulfate and phosphate transporter, SULTR3;3. We subsequently over-expressed GFP-IMM1a and IMM1a-GFP in imm1a-TA, which made the plant susceptible to disease in a dose-dependent manner (Fig. 1A). Confocal microscopy found the GFP signal on the plasma membrane of epidermal cells, including the attacked cells hosting Go haustoria (Fig. 1B).

[0314] EXAMPLE 2: immla resistance manifested at the stage of penetration and after penetration

[0315] To uncover at which step in the infection process the immla resistance is having its effect, a microscopy study was performed. While the haustorium formation rate in eds1-2 was 87%, it was 25% in imm1a-TA eds1-2. imm1a-TA single mutant has a penetration rate of 48%. Mutation in the well-known MLO2 gene only reduced the penetration rate toM& C PC932446GB

[0316] 46

[0317] 54% (Fig. 3B) in agreement with previous studies. These results agree with the finding that immla has larger papillae at Go attack sites similar to mlo2 (Fig. 3A).

[0318] Meanwhile, when a haustorium is formed in imm1a-TA, it does not support hyphal growth to the same level as a haustorium in wild-type plants. The hyphal length is only about 25% (Fig. 30). When combining the low penetration rate and the poor hyphal growth, the fungal growth becomes very low (Fig. 30). This is reflected in for instance the fungal biomass quantification by qPCR (Fig. 10).

[0319] EXAMPLE 3: immla has increased resistance to both hemibiotrophic and necrotrophic fungal pathogens.

[0320] To test whether immla knock-out confers resistance to other leaf pathogens of Arabidopsis, we inoculated imm1a-TA with spores of Hyaloperonospora arabidopsidis, Colletotrichum higginsianum and Botrytis cinerea, two hemibiotrophs and a necrotroph, respectively. In all cases, this knock-out resulted in a reduction of the disease symptoms by approximately 50% (Figures 4, 5 and 6). The mlo2-5 mutation reduced the disease symptoms of C. higginsianum and B. cinerea by approximately 40% (Fig. 4 and 5). Fungal biomass quantification showed similar results (Fig. 4 and 5).

[0321] EXAMPLE 4: No growth penalty of immla knock-out, and immla show increased grain number per plant.

[0322] It is common to observe that plant mutants with broad-range disease resistance suffer from growth penalties. To test whether this is also the case for imm1, we grew the plants under short day, followed by long day to induce flowering. Here we saw no evident change in growth (Fig. 7). In fact, seed production was increased on imm1-TA (Figure 7B).

[0323] EXAMPLE 5: Barley double-mutants show resistance to barley powdery mildew fungus, Blumeria hordei (Bh).

[0324] The single SULTR3;3 mutant (barley Ipa1-1 (= Hvimm1A-1)) was generated in the Harrington cultivar (Ye et al., 2011). Double SULTR3;3: SULTR3;4 mutants were generated via CRISPR / Cas9 mutagenesis in the Golden Promise cultivar.M& C PC932446GB

[0325] 47

[0326] As shown in Figure 15A, the single Hvimm1A-1 mutants did not show resistance to powdery mildew fungus (the mutants show the same powdery mildew colony phenotype as the Harrington wild-type). The lack of resistance is confirmed in Figure 15C - the number of colonies / cm2 in the single mutant is statistically equal to the wildtype cultivar.

[0327] As shown in Figure 15B, the Barley double mutants were resistant to powdery mildew fungus (the mutants do not display the speckled, infected phenotype lie the Golden Promise wildtype). The resistance is confirmed in Figure 15C - the number of colonies in the double mutant is significantly reduced (p<0.001) compared to the wildtype. Figure 15D shows that the mutant displays a decreased fungal penetration rate compared to the wild-type and the CRISPR-minus regenerant - thereby confirming the improved resistance.

[0328] EXAMPLE 6: MATERIALS AND METHODS

[0329] Plant material

[0330] Approximately 10,000 seed of Arabidopsis Col-0 eds1-2 (Bartsch et al., 2006) were mutagenized by shaking in 0.3% ethyl methane-sulfonate (EMS) overnight at room temperature and subsequently washed in excess tap water with several shifts to remove residual EMS. The seeds were immediately sown in the greenhouse to produce 20 separate pools of M2 mutagenized seed. Arabidopsis plants were otherwise grown at short-day (8 / 16-h photoperiod) at 21 / 15°C and 125 μE m-2s-1.

[0331] The barley Ipa1-1 (= Hvimm1A-1) mutant in cultivar Harrington was used (Ye et al., 2011). Barley CRISPR / Cas9 mutagenesis was made in cultivar, Golden Promise.

[0332] Fungal pathogens, inoculation disease scoring

[0333] The Arabidopsis powdery mildew fungus, Golovinomyces orontii (Go) isolate PMIPZ, was produced on the hypersusceptible Col-0 eds1-2. The barley powdery mildew fungus, Blumeria hordei (Bh), isolate C15, was produced on the susceptible barley line, P-02. The anthracnose fungal pathogen, Colletotrichum higginsianum, isolate IMI 349063A, was cultivated on Mathur’s medium (2.8 g glucose, 1.22 g MgSO4·7H2O, 2.72 g KH2PO4, 2.18 g Oxoid Mycological peptone, 30 g agar, per 1 liter) at 25°C in the dark. The gray mold fungal pathogen, Botrytis cinerea, isolate 2909 found on Phaseolus vulgaris in Turkey, was cultivated on potato dextrose agar.M& C PC932446GB

[0334] 48

[0335] Go and Bh inoculation of 4-week-old Arabidopsis and 1-week-old barley was made in a settling tower according to Liao et al. (2023) and the level of attack was quantified visually 10 and 6 d post inoculation (dpi), respectively. Go was as well quantified 7 dpi by qPCR of genomic DNA using the fungal ATPase 1 (Go_V1_Contig3757) gene primer pair, R193: 5’-TCGCCGCTATATTTGGAGTC-3' R194: 5’-CTGGGTCAGATGGTTCACCT-3’, relative to Arabidopsis genomic DNA using the RNA-binding (RRM / RBD / RNP motifs) family protein (At3g21215) gene primer pair, R189: 5’-GAATCCACCCATACCACCAG-3' R192: 5’- GAGGAGGAGGATGGTGATGA-3’.

[0336] C. higginsianum was spray inoculated onto leaves of 4-week-old plants and the level of attack was quantified visually 6 dpi, and at 3 dpi by qPCR of genomic DNA using the fungal actin gene primer pair, ChACT-F: 5’-CCCCAAGTCCAACAGAGAGA-3' ChACT-R: 5’-CATCAGGTAGTCGGTCAAGTCA-3’, relative to Arabidopsis genomic DNA using the actin 2 (At3g18780) gene primer pair, AtACT2-F: 5’-ATGGAAGCTGCTGGAATCCAC-3' AtACT2-R: 5’-TTGCTCATACGGTCAGCGATG-3’.

[0337] B. cinerea was drop-inoculated onto leaves of 4-week-old plants and the level of attack by B. cinerea was quantified visually 5 dpi, and at 3 dpi by qPCR of genomic DNA using the fungal cutinase A (Z69264) gene primer pair, BcCG11: 5’-AGCCTTATGTCCCTTCCCTTG-3' BcCG12: 5’-GAAGAGAAATGGAAAATGGTGAG-3’, relative to Arabidopsis genomic DNA using the a-Shaggy kinase (At5g26751) gene primer pair, ASK1: 5’-CTTATCGGATTTCTCTATGTTTGGC-3' ASK2: 5’-GAGCTCCTGTTTATTTAACTTGTACATACC-3’.

[0338] Mutant screen and IMM1 gene identification

[0339] EMS-mutagenized Col-0 eds1-2 M2 plants were grown in flats and continuously inoculated with Go by keeping Go-producing plants in the same growth chamber in front of a fan to disperse the spores. Seeds from potentially Go-resistant mutant plants were selected for further testing. Confirmed Go-resistant M3 lines were crossed to each other. When Fi‘s of such complementation test crosses were Go-resistant, the two parents were taken to carry recessive mutations in the same gene.

[0340] The Go-resistant line B1-1 was crossed to the original Col-0 eds1-2, and genomic DNA from 30 resistant F2 plants was mixed and genome sequenced with Novogene (www.novogene.com). The EMS-induced single nucleotide polymorphisms (SNPs) were mapped onto the Arabidopsis Col-0 genome and the index for each SNP was scored. An SNP in At1g23090, conferring the S371F amino acid substitution in the encodedM& C PC932446GB

[0341] 49

[0342] protein SULTR3;3, reached an index of 100%. That this mutation caused Go-resistance was confirmed 1) by sequencing this gene in other resistant mutants, for instance predicted to be allelic from the complementation tests (see Fig. 8), 2) by over-expression of a construct encoding the wild-type SULTR3;3 and observing that this reverted resistance to susceptibility, and 3) by observing that a T-DNA insertion mutation in the same gene conferred Go-resistance in its homozygous state (Fig. 1, 2 and 3).

[0343] DNA constructs

[0344] Constructs for Arabidopsis transformation were made transferring coding sequence from pENTR plasmids to 35S promotor-driven destination vectors, pK7FWG2.0 for C-terminal GFP fusion and pK7WGF2.0 for N-terminal GFP fusion, using Gateway LR Clonase (Invitrogen) reactions.

[0345] The CRISPR / Cas9 guide RNA construct for simultaneously mutation of barley IMM1A and IMM1B was made using the oligos, IMM1ABC20F: 5’-ACTTGGAGGCTGGCAATGGTGAGGCG-3’ and IMM1ABC20R: 5’- AAAACGCCTCACCATTGCCAGCCTCC-3’, which were annealed and GATEWAY™ cloned into the T-DNA destination pANIC6A vector while recombining with the entry vectors, pJG85 and pJG80, for combining sequences for sgRNA and Cas9 expression.

[0346] Plant transformation

[0347] Arabidopsis T-DNA transformation was performed by the floral dip method. Barley T-DNA transformation for CRISPR / Cas9 mutagenesis was made on immature embryos according to Holme et al. (2012).

[0348] Microscopy

[0349] Microscopic analysis of Go development and imm1-resistance in Arabidopsis was performed according to Liao et al. (2023). Confocal microscopy was performed on a Leica SP5-X laser-scanning microscope using a 63x water immersion lens and a numerical aperture of 1.20. The AtIMM1a-GFP fusion protein was excited at 514 nm while emission was collected in the range, 527-586 nm.

[0350] References

[0351] Bartsch M, Gobbato E, Bednarek P, Debey S, Schultze JL, Bautor J, Parker JE. 2006. Salicylic acid-independent ENHANCED DISEASE SUSCEPTIBILITY1 signaling inM& C PC932446GB

[0352] 50

[0353] Arabidopsis immunity and cell death is regulated by the monooxygenase FMO1 and the Nudix hydrolase NUDT7. Plant Cell 18, 1038-1051.

[0354] Ding G, Lei GJ, Yamaji N, Yokosho K, Mitani-Ueno N, Huang S, Ma JF. (2020) Vascular Cambium-Localized AtSPDT Mediates Xylem-to-Phloem Transfer of Phosphorus for Its Preferential Distribution in Arabidopsis. Mol. Plant 13, 99-111.

[0355] Sunzhenhe Fang et al. (2025) Molecular mechanism underlying phosphate distribution by SULTR family transporter SPDT in Oryza sativa. Sci. Adv.11,eady3442, DOI: 10.1126 / sciadv.ady3442

[0356] Gu M, Huang H, Hisano H, Ding G, Huang S, Mitani-Ueno N, Yokosho K, Sato K, Yamaji N, Ma JF (2022) A crucial role for a node-localized transporter, HvSPDT, in loading phosphorus into barley grains. New Phytol. 234, 1249-1261.

[0357] Holme IB, Brinch-Pedersen H, Lange M, Holm PB (2012) Transformation of barley (Hordeum vulgare L.) by Agrobacterium tumefaciens infection of In Vitro cultured ovules. Methods in Molecular Biology 847, 151-161.

[0358] Liao W, Nielsen MW, Pedersen C, Xie WJ, Thordal-Christensen H (2023) Barley endosomal MONENSIN SENSITIVITY1 is a target of the powdery mildew effector CSEP0162 and plays a role in plant immunity. Journal of Experimental Botany 74, US-129.

[0359] Ye H, Zhang XQ, Broughton S, Westcott S, Wu D, Lance R, Li C. (2011) A nonsense mutation in a putative sulphate transporter gene results in low phytic acid in barley. Funct. Integr. Genomics. 11, 103-110.

[0360] Fatima U, Okla MK, Mohsin M, Naz R, Soufan W, Al-Ghamdi AA, Ahmad A. A Non-Invasive Tool for Real-Time Measurement of Sulfate in Living Cells. Int J Mol Sci. 2020 Apr 7;21(7):2572. doi: 10.3390 / ijms21072572. PMID: 32272790; PMCID: PMC7177696.

[0361] Sequence Listing

[0362] Materials and methods sequencesM& C PC932446GB

[0363] 51

[0364] SEQ ID NO: 57 - TCGCCGCTATATTTGGAGTC

[0365] SEQ ID NO: 58 - CTGGGTCAGATGGTTCACCT

[0366] SEQ ID NO: 59 - GAATCCACCCATACCACCAG

[0367] SEQ ID NO: 60 - GAGGAGGAGGATGGTGATGA

[0368] SEQ ID NO: 61 - CCCCAAGTCCAACAGAGAGA

[0369] SEQ ID NO: 62 - CATCAGGTAGTCGGTCAAGTCA

[0370] SEQ ID NO: 63 - ATGGAAGCTGCTGGAATCCAC

[0371] SEQ ID NO: 64 - TTGCTCATACGGTCAGCGATG

[0372] SEQ ID NO: 65 - AGCCTTATGTCCCTTCCCTTG

[0373] SEQ ID NO: 66 - GAAGAGAAATGGAAAATGGTGAG

[0374] SEQ ID NO: 67 - CTTATCGGATTTCTCTATGTTTGGC

[0375] SEQ ID NO: 68 - GAGCTCCTGTTTATTTAACTTGTACATACC

[0376] SEQ ID NO: 69 - ACTTGGAGGCTGGCAATGGTGAGGCG

[0377] SEQ ID NO: 70 - AAAACGCCTCACCATTGCCAGCCTCC

[0378] SEQ ID NO: 75 - GAGGCTGGCAATGGTGAGGC

[0379] Amino Acid Sequences

[0380] • SULTR 3;3 chosen from: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56 • SULTR 3;4 chosen from: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56

[0381] SEQ ID NO: 1 > AtlMM1A (SULTR3;3=AT1G23090). Accession number: NP_173722.1. Serine124 in bold and underlined. This is the corresponding position to Serine170 in Fang et al., shown to be important for substrate specificity of the transporter.

[0382] MEVHKVVAPPHKSTVAKLKTKLKETFFPDDPLRQFRGQPNRTKLIRAAQYIFPILQWC PEYSFSLLKSDVVSGLTIASLAIPQGISYAKLANLPPIVGLYSSFVPPLVYAVLGSSRDLA VGPVSI ASLI LGSM LRQQVSPVDDPVLFLQLAFSSTFFAGLFQASLGI LRLGFI I DFLSKA TLIGFMGGAAIIVSLQQLKGLLGITHFTKHMSVVPVLSSVFQHTNEWSWQTIVMGVCFL LFLLSTRHLSMKKPKLFWVSAGAPLLSVIVSTLLVFVFRAERHGISVIGKLPEGLNPPS WNMLQFHGSHLALVAKTGLVTGIVSLTEGIAVGRTFAALKNYHVDGNKEMIAIGLMNV VGSATSCYVTTGAFSRSAVNNNAGAKTAVSNIVMSVTVMVTLLFLMPLFEYTPNVVLG AIIVTAVIGLIDLPAACHIWKIDKFDFLVMLCAFFGVIFLSVQNGLAIAVGLSLFKILMQVT RPKMVIMGNIPGTDIYRDLHHYKEAQRIPGFLVLSIESPVNFANSNYLTERTSRWIEEC EEEEAQEKHSSLQFLILEMSAVSGVDTNGVSFFKELKKTTAKKDIELVFVNPLSEVVEK LQRADEQKEFMRPEFLFLTVAEAVASLSLKGPSLSNVM& C PC932446GB

[0383] 52

[0384] SEQ ID NO: 2 > AtlMM1B (SULTR3;4=AT3G 15990) Serine147 in bold and underlined. This is the corresponding position to Serine170 in Fang et al., shown to be important for substrate specificity of the transporter.

[0385] MGHGTNRVEDMASPNNGTAGETWEIHSVCLPPKKTAFQKLKKRVGDVFFPDDPLQ RFRNQTWRNRVILGLQSLFPIFTWGSQYDLKLLRSDVISGLTIASLAIPQGISYAKLANL PPIVGLYSSFVPPLIYAVLGSSRHLAVGPVSIASLVMGSMLSESVSPTQDSILYLKLAFT STFFAGVFQASLGLLRLGFMIDFLSKATLIGFTAGAAVIVSLQQLKGLLGIVHFTGKMQI VPVMSSVFNHRSEWSWETIVMGIGFLSILLTTRHISMRKPKLFWISAASPLASVIISTLL VYLIRSKTHAISFIGHLPKGLNPPSLNMLYFSGAHLALAIKTGIITGILSLTEGIAVGRTFA SLKNYQVNGNKEMMAIGFMNMAGSCTSCYVTTGSFSRSAVNYNAGAKTAVSNIVMA SAVLVTLLFLMPLFYYTPNVILAAIILTAVIGLIDYQAAYKLWKVDKFDFFTCLCSFFGVLF VSVPLGLAIAVAVSVIKILLHVTRPNTSEFGNIPGTQIYQSLGRYREASRIPGFLILAIESPI YFANSTYLQDRILRWAREEENRIKENNGTTLKCIILDMTAVSAIDTSGLEAVFELRRRLE KQSLQLVLVNPVGTVMEKLHKSKIIEALGLSGLYLTVGEAVADLSSTWKANGQP

[0386] Hordeum vulgare (barley)

[0387] SEQ ID NO: 3 > HvlMM1A (SULTR 3;3) HORVU. MQREX.r3.2HG0204020.1 Serine170 in bold. This is the corresponding position to Serine170 in Fang et al., shown to be important for substrate specificity of the transporter.

[0388] MVGMSGAYGGHHGNGGEGRAAPKQAWVGAPPPPAAAEMEMEIGVVHKVAAQPA QSTASKMKGKVKETFFPDDPFRSFKGQPVRAQWVLAAKYLFPVLEWVPGYSLSLFKS DLVAGLTIASLAIPQGISYAKLANLPPIIGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLIM GSMLRQAVSPSASPALFLQLAFTSTFFAGLVQASLGILRLGFIIDFLSKATLVGFMAGA AIIVSLQQLKALLGIVHFTTQMGIVPVMASVFQHTNEWSWQTILMGACFLVLLLAARHV SMRWPKFFWISACAPLASVIVSTLLVFLFKAQNHGISIIGSLKCGLNRPSWDQLLFDTT YLGLTMKTGLVTGIISLTEGVAVGRTFASLKDYQVDGNKEMMAIGLMNIVGSCTSCYV TTGAFSRSAVNHNAGCKTAMSNWMALTVMVTLLFLMPLFVYTPNVVLGAIIIAAVIGLI DLPAAYNIWKMDKMDFLVCLCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPRMMIQGN IKGTDIYRNLHQYKEAQRVPGFLILTIEAPINFANTNYLNERTKRWIEDESFSGNKQSEL RVVILDLSAVPAIDTSGIAFLIDLKKSTEKHGLELVLVNPTGEVMEKIQRANDAHNHFRQ DCLYLTTGEAIASLSGFAKMATP

[0389] SEQ ID NO: 4 > HvlMM1C (SULTR 3;3) HQRVU. MQREX.r3.7HG0741010.1. Prospective pseudogene

[0390] Threoninel 11 (a polar, uncharged amino acid like serine) in bold - this is the equivalent position to Serine170 described in Fang et al.M& C PC932446GB

[0391] 53 MEGTSHAYGGHNGSEILPPRQGMWGAPRPAEVSPVGMVHKVMAQPAQSTASKMK GKVKETFFPDDPFRSFKGQPLRKKWLMAAKYLLPSVEWVPGYSLSLFRSDLIAGFTIA SVAIPQGISYAKLADLPPIIGLYSSFVPPLVYAVLGSSHDLAVGPTSITSLIMGSMLQKAV LVSPTAEPALFVQLAFTSTLFAGLLQASLGILRLGFIIDFLSKATLLGFMAGAAIIVSLQQL KELLGIIHFTDKMDLVDVMASVFQHTDEWSWQTILMGACFLVLLLSARHVSMRWPKF FWISACAPLVSIIMSTVLVFIFKAENHGISVIGHIKCGLNHLSWDKLLFDPKYLGLAMKT G LVTGI I SLTEGVAVG RTFASI KDYQVDG N KEM M Al G LM N I VGSCTSCYVTTGG FSRS AVNHNAGCKTAMSNVIMALTVMVTLLFLMPLFVYTPNVVLGAIITVAVVGLIDVPAAYHI WKMDKMDFLVCLCAFAGVIFISVEEGLAIAVGISIFRVLMQITRPRMIIQGNIKGTDIYRNI HQYEEAQRVPGFLILTVEAPINFANTNYLNERTKRWIEDESFSRNKSELRFVIFDLSAV PAIDTSGIAFLVDLKKPTEKLGLELVLVNPTGEVMEKIQRANDPHNHFRPDCLYLTIGEA IASLSGEANMATP

[0392] SEQ ID NO: 5 > HvlMM1B (SULTR 3;4) HORVU. MOREX.r3.7HG0648840.1 =HvSPDT Serine160 in bold - This is the corresponding position to Serine170 in Fang et al., shown to be important for substrate specificity of the transporter.

[0393] MVVNNKVETLAFDVEAGQGQGPRPKGAAESGAAGRVVELHKVSAPERRTTCRALGQ RLAEIFFPDDPLHQFKNQSLARKLVLALQYFFPIFHWGSNYSLRLLRSDAVAGLTIASL AIPQGISYAKLANLPPIIGLYSSFVPPLIYALLGSSRDLAVGPVSIASLVMGSMLREAVAP EQQPILYLQLAFTATFFAGLFQASLGFLRLGFIVDFLSKATLTGFMGGAAVIVSLQQLK GLLGIVHFTTHMGFVDVMASWRRHSEWEWQTIVMGVAFLAILLGTRQISARNPRLF WVSAAAPLTSVIASTIISYLCRGHAISIIGDLPRGVNPPSMNMLVFSGSYVALAIKTGIMT GILSLTEGIAVGRTFASINNYQVDGNKEMMAIGVMNMAGSCASCYVTTGSFSRSAVN YSAGCRTAVSNIVMAAAVLVTLLFLMPLFHYTPNVILSAIIITAVAGLIDVRGAAKLWKVD KLDFCACVAAFLGVLLVSVQVGLAVAVGISLFKILLQVTRPNTVVMGLVPGTQSYRSM AQYREAVRVPPFLVVGVESAIYFANSTYLVERIMRYLREEEERAAKANLCGVRCIVLD MSAVTAIDTSGLDALAEMKRVLDKRGIDLVLANPVGSVTERMYNSVVGDTFGSGRIFF SVDEAVAAAPYKAQP

[0394] Oryza sativa

[0395] SEQ ID NO: 6 > OsLPA1 Qs04t0652400 (SULTR 3;3)

[0396] Serine157 in bold - this corresponds to Serine170 of OsSPDT, described in Fang et al. MVGMRGAYGYNDNGNEGGWVGGGEAEAEIAAMAVLHKVAAPPAQSTASKLKARV KETFFPDDPFRGFKGKPLTTKWVMAVQYLFPILDWVPSYSFSLFKSDLVAGLTIASLAI PQGISYAKLASLPPIIGLYSSFVPPMVYAVLGSSRDLAVGPVSIASLIMGSMLRQAVSP AAEPLLFLQLAFTSTFFAGLVQASLGI LRLGFI I DFLSKATLVGFMAGAAI I VSLQQLKALLM& C PC932446GB

[0397] 54 GIVHFTTEMGLVPVMASVIHHTKEWSWQTILMAVCFLVLLLTARHVSMKWPKLFWVS ACAPLACVIVSTLLVFLFKAQKHGISIIGQLKCGLNRPSWDKLLFDPQYLGLTVKTGLVT

[0398] G 11 SLTEGVAVG RTFASLKDYQVDG N KEM M Al G LM N I VGSCTSCYVTTGAFSRSAVN H NAGCKTAMSNVIMALTVMVTLLFLMPLFVYTPNVVLGAIIIAAVIGLIDLPAVYNIWKMDK MDFLVCLCAFAGVIFISVQQGLAIAVGISIFRVLLQITRPKMMIQGNIKGTDIYRNLHQYK DAQRVPGFLILTVEAPINFANTNYLNERIKRWIEEESSAGTKQSELHFVILDLSAVPAIDT SGISFLIDLKKSTEKHGLELILVNPTGEVMEKIQRANDAHGHFKSDSLYLTTGEAVASLS TFSKMTAP

[0399] SEQ ID NO: 7 >0s06t0143700 =OsSPDT (SULTR 3;4) Serine170 of Fang et al. in bold. MVVNNKVDSLSYDVEAPPAQAPTTPAVVSAPPTPRGEAPAMTTTAAAELHKVSVPER RSTAKALRQRLAEVFFPDDPLHQFKNQSSARRLVLALQYFFPIFHWGSDYSLRLLRSD VVSGLTIASLAIPQGISYAKLANLPPIIGLYSSFVPPLIYSLLGSSRDLAVGPVSIASLVMG SMLRQAVSPDQEPILYLQLAFTSTFFAGVFQASLGFLRLGFIVDFLSKATLTGFMGGAA

[0400] 11 VSLQQLKGLLGI I H FTSQMGFVQVM HSVFKH H DEWAWQTI LMGVAFLAVLLTTRH IS ARNPKLFWVSAAAPLTSVIISTIISFVSKAHGISVIGDLPKGLNPPSANMLTFSGSYVGL ALNTGIMTGILSLTEGIAVGRTFASINNYQVDGNKEMMAIGVMNMAGSCASCYVTTGS FSRSAVN YSAGCKTAVSN I VMASAVLVTLLFLM PLFHYTPN VI LSAI I ITAVIGLI DVRGAA RLWKVDKLDFLACMAAFLGVLLVSVQMGLAIAVGISLFKILLQVTRPNMVVKGVVPGT ASYRSMAQYREAMRVPSFLVVGVESAIYFANSMYLGERIMRFLREEDERAAKCNQCP VRCIILDMSAVAAIDTSGLDALAELKKVLEKRNIELVLANPVGSVTERLYNSVVGKTFGS DRVFFSVAEAVAAAPHKTQP

[0401] Brassica napus

[0402] SEQ ID NO: 8 > CDY25768 SULTR 3;3 orthologue MEVHKVVAPPHRSTAAKLKTRLKETFFPDDPLRQFKGQPNRTKLIRAAQYIFPILQWC PEYSFRLLKSDWSGLTIASLAIPQGISYAKLANLPPIVGLYSSFVPPLVYAVLGSSRDL A GPVSIASLILGSMLRQQVSPVDNPLLFLQLAFSSTFFAGLFQASLGILRLGFIIDFLSK ATLIGFMAGAAIIVSLQQLKALLGITHFTKQMGVVPVLSSVFHHTNEWSWQTIVMGVCF LLFLLATRHLSMKKPKLFWVSAGAPLLSVIVSTLLVFVFRADRHGISVIGKLQEGLNPPS WNMLQFHGSHLGLVAKTGLITGIVSLTEGIAVGRTFAALKNYHVDGNKEMIAIGLMNVV GSATSCYVTTGAFSRSAVNNNAGCKTAVSNI MSVTVMVTLLFLMPLFEYTPNVVLGA IIVTAVIGLIDLPAARHIWRIDKFDFLVMLCAFFGVIFLSVQQGLAIAVGLSLFKLLMQVTR PKTVIMGNIPGTDIYRNLHHYKEARRIPGVLVLSIESAVNFANSNYLTERTSRWIEDSEEM& C PC932446GB

[0403] 55 EEAQEKHSSLQFLILEMSAVSGVDTNGVSFFKELKKTTAKKNIELVFVNPLSEVMEKLQ RADEEEEFMRPEFLFLTVAEAVASLSLKGPSLNNV

[0404] SEQ ID NO: 9 > CDY21415 SULTR 3;3 orthologue MEVHKVVAPPHRSTAAKLKTRLKETFFPDDPLRQFRGQPNRTKLIRAAQYIFPILQWC PEYSFRLLKSDWSGLTIASLAIPQGISYAKLANLPPIVGLYSSFVPPLVYAVLGSSRDL AVGPVSIASLILGSMLRQQVSPVDNPLLFLQLAFTSTFFAGLFQASLGILRLGFIIDFLSK ATLIGFMAGAAIIVSLQQLKALLGITHFTKQMSVIPVLSSVFHHTNEWSWQTIVMGVCFL LFLLATRHLSMKKPKLFWVSAGAPLLSVILSTLIVFVSRADRHGISVIGKLQEGLNPPSW NMLQFHGSHLGLVAKTGLITGIVSLTEGIAVGRTFAAVKNYHVDGNKEMIAIGLMNVVG SATSCYVTTGAFSRSAVNNNAGCKTAVSNIVMSVTVMVTLLFLMPLFEYTPNWLGAII VTAVIGLIDLPAARHIWRIDKFDFLVMLCAFFGVVFLSVQNGLAIAVGLSLFKLLMQVTR PKTVVMGNIPGTDVYRNLHHYKDAQRIPGFLVLSIESPVNFANSNYLTERTSRWIEEC EEEEAQEKHSSLRFLILEMSAVSGVDTNGVSFFKELKKTTAKKNIELVFVNPLSEVMEK LQRADEEEEFMRPEFLFLTVSEAVASLSLKGGPSFNNV

[0405] SEQ ID NO: 10 > CDY04744 SULTR 3;3 orthologue MEVHKVVAPPHRSTAAKLKTRLKETFFPDDPLRQFRGQPNRTKLIRAAQYIFPILQWC PEYSFRLLKSDWSGLTIASLAIPQGISYAKLANLPPIVGLYSSFVPPLVYAVLGSSRDL AVGPVSIASLI LGSM LRQQVSPVDN PI LFLQLAFTSTFFAGLFQASLGVLRLGFI I DFLSK ATLIGFMAGAAIIVSLQQLKALLGITHFTKQMSVIPVLSSVFHHTNEWSWQTIVMGVCFL LFLLATRHLSMKKPKLFWVSAGAPLLSVIVSTLIVFVSRADRHGISVIGKLQEGLNPPS WNMLQFHGSHLGLVAKTGLITGIVSLTEGIAVGRTFAAVKNYHVDGNKEMIAIGLMNV VGSATSCYVTTGAFSRSAVNNNAGCKTAVSNIVMSVTVMVTLLFLMPLFEYTPNVVLG AIIVTAVIGLIDLPAARHIWRIDKFDFLVMLCAFFGVVFLSVQNGLAIAVGLSLFKLLMQV TRPKTVVMGNIPGTDVYRNLHHYKEAQRIPGFLVLSIESPVNFANSNYLTERTSRWIEE CEEEEAQEKHSSLRFLILEMSAVSGVDTNGVSFFKELKKTIAKKRIELVFVNPLSEVME KLQRADEEEEFMRPEFLFLTVSEAVASLSLKGGPYLNNV

[0406] SEQ ID NO: 11 > CDY20171 SULTR 3;4 orthologue MGHGTNRVEDMASPNNGTTAPATANARETIVEIHSVCLPPKKTAFQKLKKRFGDVFF PDDPLERFRNQTWRNRVILCLQSLFPIFTWGSQYDLKLLRSDVVSGLTIASLAIPQGIS YAKLANLPPIVGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGSMLSESVSPTQDSIL YLKLAFTSTFFAGLFQASLGLLRLGFVIDFLSKPTLIGFTAGAAVIVSLQQLKGLLGIVHF TGKMQFIPVMSSVFSHRSEWSWETILMGLGFLAILLTTRHISMRKPKLFWISAASPLASM& C PC932446GB

[0407] 56 VVISTLLVFLIRNKTHAISFIGHLPKGLNPPSSNMLYFSGTHLALAIKTGIITGILSLTEGIA VGRTFASLKNYQVNGNKEMMAIGFMNMAGSCTSCYVTTGSFSRSAVNYNAGAKTAV SNIVMASAVLVTLLFLMPLFYYTPNLILAAIILTAVIGLIDYQAAYKLWKVDKFDFFTCMC SFFGVLLVSVPLGLAIAVGVSVLKILLHVTRPNTLEFGNIQGTQIYQSVKRYREASRIPG FLI LAVESPI YFANSTYLQERI LRWTREEEARI KEN NGSTLKCI I LDMTAVSSI DTNGI EAL FELRRRLEKQSLQLVLVNPVGSVMEKLHKSKIIESLGLSGLYLTVGEAVADLSSTWKA HGQP

[0408] SEQ ID NO: 12 > CDX82421 SULTR 3;4 orthologue MGHGTNRVEDMTSPNTATARETIVEIHSVCLPPKKTTFQKLKKRFADVFFPDDPLERF RNQTWRNKVILGLQSLFPIFTWGSQYDLKLFRSDVISGLTIASLAIPQGISYAKLANLPPI VGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGSMLSESVSPTQDPVLYLKLAFTST FFAGLFQASLGLLRLGFLIDFLSKPTLVGFTAGAAVIVSLQQLKGLLGIVHFTGKMQFIP VMSSVFNHRSEWSWETIVMGVGFLIILLTTRHISMRKPKLFWISAASPLASVVISTLLVF LIRNKTHAISFIGHLPKGLNPPSSNMLYFSGTHLALAIKTGIITGVLSLTEGIAVGRTFASL KNYQVNGNKEMMAIGFMNMVGSCTSCYVTTGSFSRSAVNYNAGAKTAASNIVLASTV LVTLLFLM PLFYYTPN LI LAAI I LTAVIGLI DYQAAYKLYKVDKFDFFTCMCAFFGVLLVSV PLGLAIAVWSVIKILLHVTRPNTLEFGNIQGTQIYQSLKRYREASRIHGFLILAVESPIY FVNSTYLQERILRWTREEETRIKENNGTTLKCIVLDMTAVSSIDTSGIEAVFELRRRLEK QSLQLVLVNPVGSVMEKLQKSKIIESLGLSGLYLTVGEAVADLSSTWKAHGQP

[0409] SEQ ID NO: 13 > CDY30127 SULTR 3;4 orthologue MGHGTNRIEDMSSPNNETAANARETWEIHSVCLPPKKTTFQKLKKRFGDVFFPDDPL ERFRNQTWRNKVILGLQSLFPIFTWGSQYDLKLFRSDVISGLTIASLAIPQGISYAKLAN LPPIVGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGSMLSESVSPTQDSILYLKLAF TSTFFAGLFQASLGLLRLGFAIDFLSKATLVGFTAGAAVIVSLQQLKGLLGIVHFTGKMQ FVPVMSSVINTRSEWSWETIVMGLGFLIILLTTRHISMRKPKLFWISAASPLASVVISTLL VYVIRDKTHAISFIGHLPKGLNPPSANMLYFSAAHLALAIKTGIITGILSLTEGIAVGRTF ASLKNYQVNGNKEMMAIGFMNMAGSCTSCYVTTGSFSRSAVNVNAGAKTAVSNIVM ASAVLVTLLFLMPLFYYTPNLILAAIILTAVIGLIDYQAAYKLWKVDKFDFFTCMCSFFGV LFVSVPLGLAIAVGVSVIKILLHVTRPNTLEFGNIPETQIYQSLKRYREASRIPGFLILAVE SPIYFANCTYLQERISRWTREEENRIKENNERNLKCIILDMTAVSSIDTSGIESVFELRR RLEKQSLQLVLVNPVGSVMEKLHKSKIIESLGLSGLYLTVGEAVSDLSSTWKAHGQP

[0410] SEQ ID NO: 14 > CDX75862 SULTR 3;4 orthologueM& C PC932446GB

[0411] 57 MGHGTNRVEDMTSPNTATARETIVEIHSVCLPPKKTTFQKLKKRFADVFFPDDPLERF RNQTWRNKVILGLQSLFPIFTWGSQYDLKLFRSDVISGLTIASLAIPQGISYAKLANLPPI VGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGSMLSESVSPTQDPVLYLKLAFTST FFAGLFQASLGLLRLGFLIDFLSKPTLVGFTAGAAVIVSLQQLKGLLGIVHFTGKMQFIP VMFSVFNHRSEWSWETIVMGLGFLIILLTTRHISMRKPKLFWISAASPLASVVISTLLVF LIRNKTHAISFIGHLPKGLNPPSSNMLYFSGTHLALAIKTGIITGVLSLTEGIAVGRTFASL KNYQVNGNKEMMAIGFMNMVGSCTSCYVTTGSFSRSAVNYNAGAKTAASNIVLASTV LVTLLFLM PLFYYTPN LI LAAI I LTAVIGLI DYQAAYKLYKVDKFDFFTCMCAFFGVLLVSV PLGLAIAVVVSVIKILLHVTRPNTLEFGNIQGTQIYQSLKRYREASRIHGFLILAVESPIY FANSTYLQERILRWTREEETRIKENNGSTLKCIVLDMTAVSSIDTSGIEAVFELRRRLEK QSLQLVLVNPVGSVMEKLHKSKIIESLGLSGLYLTVAEAVADLSSTWKAHGQP

[0412] SEQ ID NO: 15 > CDX97715 SULTR 3;4 orthologue MGHGTNRIEDMSSPNNETAANARETWEIHSVCLPPKKTTFQKLKKRFGDVFFPDDPL ERFRNQTWRNKVILGLQSLFPIFTWGSQYDLKLFRSDVISGLTIASLAIPQGISYAKLAN LPPIVGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGSMLSESVSPTQDSILYLKLAF TSTFFAGLFQASLGLLRLGFVIDFLSKATLVGFTAGAAVIVSLQQVKGLLGIVHFTGKM QFVPVMSSVINTRSEWSWETIVMGLGFLIILLTTRHISMRKPNLFWISAASPLASWIST LLVYLIRDKTHAISFIGHLPKGLNPPSVNMLYFSAAHLALAIKTGIITGILSLTEGIAVGRTF ASLKNYQVNGNKEMMAIGFMNMAGSCTSCYVTTGSFSRSAVNVNAGAKTAVSNILM ASAVLVTLLFLMPLFYYTPNLILAAIILTAVIGLIDYQAAYKLWKVDKFDFFTCMCSFFGV LFVSVPLGLAIAVGVSVIKILLHVTRPNTVEFGNIPGTQIYQSLKRYREASRVPGFLILAV ESPLYFANCTYLQERILRWTREEENRIKENNDRNLKCIILDMTAVSSIDTSGIEAVFELR RRLEKQSLQLVLVN PVGSVM EKLH KSKI I ESLGLSGLYLTVGEAVSDLSSTWKAHGQP

[0413] Cucumis sativus (cucumber)

[0414] SEQ ID NO: 16 > KGN50848 MGDDDNNNNNNRNGSSKQSEPPRGPPEHSVEIIHPVVPPPRRSWLEKIRNRLKEIFF PDDPLRQFKGQSPVRKLVLGAQYI FPI LEWGSHYN FSLFKSDVVAGLTI ASLAI PQGIS YAKLANLPPIVGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLILGSMLRQEVSPIKDPILF LQLGFTATFFAGLFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKGLLGITHFTK QMGLIPVLSSVFHHTHEWSWQTILMGFCFLLFLLLTRHISMKRPKLFWVSAGAPLVSVI LSTILVFAFKADRHGISIIGKLEEGLNPPSLNMLRFEGSHLGLVIKTGLVTGIISLTEGIAV GRTFAAIKDYRVDGNKEMIAIGLMNVVGSFTSCYVTTGAFSRSAVNHNAGAKTAVSNI VMSVTIMVTLLFLMPLFQYTPNLVLAAIIVTAVIGLIDVPAAYAIWKVDKFDFVVMLCAFFM& C PC932446GB

[0415] 58 GVILISVQHGLAIAVGISIFKIILQITRPKTAMLGNIGGTDIYRNIHQYKDAMSIQGFLILSIE APINFANTTYLNERILRWIEDYEAGQDHLKKEGSDLQFVVLELSAVSAIDTSGVLLFKDL RRALEKKGVELVLVNPMGELLEKLQKADENQEILRPNNVFLTVGEAVAFLSATMKRQ SSTI

[0416] SEQ ID NO: 17 > KGN45350 MGINSNRVENLECRETVLTMPADAMPEPSRPEIEIHKVCLPPEQTTFQKLKHKLSEVF FPDDPFHRFKNQTTLRKLLLGLQFLFPVFQWGPEYTLALFKSDWSGLTIASLSIPQGI SYAKLANLPPIIGLYSSFVPPLIYSILGSSRHLAVGPVSIASLVMGSMITEAVSYNEHPTL YLKLAFTATFFAGVFQASLGLLRLGFVIDFLSKATLVGFMAGAAVIVSLQQFKGLLGIAH FTTKMQFIPVMSSVFHRKDEWSWQTIVLGFIFLLFLLGTRHISIKKPKLFWISAAAPLTS VILSTILVFLLRTKFPGISVIGHLPKGVNPPSLNMLYFTGPQLVLAIKTGIITGILSLTEGI AVGRTFAGLKNYQVDGNKEMMAIGFMNVAGSCSSCYVTTGSFSRSAVNYNAGAQTA VSNVVLSAAVLITLLFLMPLFHYTPNFILAAIIITAVIGLIDYQAACKLWKVDKLDFLACVC SFFGVLFISVPLGLAIAVGVSVFKILLHVTRPNTMVLGNISGTHIFQNLDRYRDASRVPS FLILAIDSPIYFANSTYLQERILRWVREEEERIKSTEDSPLKCVILDMTAVTSIDTSGIETV CELKKILMKKSLQFVLANPGGNVMEKLYNSKALEQFEFNGLYLSVGEAVKDISSLWKR PLS

[0417] Avena sativa (oat)

[0418] SEQ ID NO: 18 MVGMSGAYGGHNGSESRQTKEAVVVGAPEMGVVHKVASPPAQSTASKMKGKVKET FFPDDPFRSFKGQPQRAKLLLAVKYLFPVLEWAPGYTLSLFKSDLVAGLTIASLAIPQGI SYAKLANLPPIIGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLIMGSMLRQAVSPSAEPA LFLQLAFTSTLFAGLVQASLGILRLGFIIDFLSKATLVGFMAGAAVIVSLQQLKALLGIVH FTTQMGIVPVMASVFQHTDEWSWQTILMGACFLVLLLTARHVSIRWPKFFWISACAPL ACVIVSTLLVFLFKAQNHGISIIGQLKCGLNRPSWDKLLFDPTYLGLTMKTGLVTGIISLT EGVAVGRTFASLKDYQVDGNKEMMAIGLMNVVGSCTSCYVTTGAFSRSAVNHNAGC KTAMSNVIMALTVMVTLLFLMPLFVYTPNWLGAIIIAAVIGLIDLPAAYNIWKMDKMDFL VCLCAFAGVIFISVQQGLAIAVGISIFRVLMQITRPRMMIQGNIKGTDIYRNLHQYKEAQ RVPGFLI LTVEAPI N FANTNYLN ERTKRWI EDESSSGN KQTELH FVI LDLSAVPAI DTSGI AFLIDLKKATEKRGLELVLVNPTGEVMEKIQRANDAHDHFRPDCLYLTTGEAIASLSGF AKMATP

[0419] SEQ ID NO: 19M& C PC932446GB

[0420] 59 MVGMSGAYGGHNGSESRQSKEAVVVGAPEMIGVGLGMGWHKVASPPAQSTASKM KGKVKETFFPDDPFRSFKGQPQRAKLLLAVKYLFPVLEWAPGYTLSLFKSDLVAGLTI ASLAIPQGISYAKLANLPPIIGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLIMGSMLRQA VSPSAEPALFLQLAFTATLFAGLVQASLGILRLGFIIDFLSKATLVGFMAGAAVIVSLQQL KALLGIVHFTTQMGIVPVMASVFQHTDEWSWQTILMGACFLVLLLTARHVSIRWPKFF WISACAPLACVIVSTLLVFLFKAQNHGISIIGQLKCGLNRPSWDKLLFDPAYLGLTMKTG LVTGIISLTEGVAVGRTFASLKDYQVDGNKEMMAIGLMNVVGSCTSCYVTTGAFSRSA VNHNAGCKTAMSNVIMALTVMVTLLFLMPLFVYTPNVVLGAIIIAAVIGLIDLPAAYNIWK MDKMDFLVCVCAFAGVIFISVQQGLAIAVGISIFRVLMQITRPRMMIQGNIKGTDIYRNL HQYKEAQRVPGFLI LTVEAPI N FANTNYLN ERTKRWI EDESSSGN KQTELH FVI LDLSA VPAIDTSGIAFLIDLKKATEKRGLELVLVNPTGEVMEKIQRANDAHDHFRPDCLYLTTG EAIASLSGFAKMATP

[0421] Gossypium raymondii (Cotton)

[0422] SEQ ID NO: 20 > KJB15440 MEYSNSSNTEQPQTCLEITMEVHRVVPPPHKSTIHKLKTTLKETFFPDDPLRQFKGQP TGKKWILAAQYIFPILQWGPHYSLGLFKSDIVAGLTIASLAIPQGISYAKLANLPPIVGLY SSFVPPLVYAVLGSSRDLAVGPVSIASLILGSMLRQEVSPVSDPVLFLQLAFTTTFFAG LFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKSLLGITHFTTKMGLVPVLSSVF HNTEEWSWQTILMGFCFLVFLLVARHVSMKRPKLFWVSAGAPLVSVILSTLLVFAFKA QHHGISIIGKLQEGLNPPSWNMLQFRGSHLGLSIKTGLVTGIISLTEGIAVGRTFAALKN YKVDGNKEMMAIGLMNMVGSSTSCYITTGAFSRSAVNHNAGAKSAVSNIVMSITVMV TLLFLMPLFQYTPNWLGAIIVSAVVGLIDIPAACQIWKIDKFDFIVMLCAFFGVIFISVQD GLAMAVGMSIFKILLQITRPKTVMLGNIPGTDIFRDLHHYKESMKIPGFLILSIEAPINFAN STYLNERILRWIEEYEAEDHKKQSSLQFVVLVMSGKQSQPWKQSNDAGICG

[0423] SEQ ID NO: 21 > KJB15439 MEYSNSSNTEQPQTCLEITMEVHRVVPPPHKSTIHKLKTTLKETFFPDDPLRQFKGQP TGKKWILAAQYIFPILQWGPHYSLGLFKSDIVAGLTIASLAIPQGISYAKLANLPPIVGLY SSFVPPLVYAVLGSSRDLAVGPVSIASLILGSMLRQEVSPVSDPVLFLQLAFTTTFFAG LFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKSLLGITHFTTKMGLVPVLSSVF HNTEEWSWQTILMGFCFLVFLLVARHVSMKRPKLFWVSAGAPLVSVILSTLLVFAFKA QHHGISIIGKLQEGLNPPSWNMLQFRGSHLGLSIKTGLVTGIISLTEGIAVGRTFAALKN YKVDGNKEMMAIGLMNMVGSSTSCYITTGAFSRSAVNHNAGAKSAVSNIVMSITVMV TLLFLMPLFQYTPNWLGAIIVSAVVGLIDIPAACQIWKIDKFDFIVMLCAFFGVIFISVQDM& C PC932446GB

[0424] 60 GLAMAVGMSIFKILLQITRPKTVMLGNIPGTDIFRDLHHYKESMKIPGFLILSIEAPINFAN STYLNERWIEEYEAEDHKKQSSLQFVVLVMSAVSAIDTSGVSIFKELKKTVEKKGAELV LVNPLGEVMEKLQKSDEAGDFMRPDCLFLTVGEAVATLTATIKSQVSNHVV

[0425] SEQ ID NO: 22 > KJB50368 MESNPSTMQEQSPSCLEITMEVHSWPPPHKSTIQKLKTRLKETLFPDDPFRQFKGQP TKKKWVLAAQYIFPILHWGPNYNLKLFKSDIVSGVTIASLAIPQGISYAKLASLPPIVGLY SSFVPPLVYAVLGSSRDIAVGPVSIASLILGSMLTQQVSPTGDPLLFLQLAFTATFFAGL FQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKSLLGITHFTKKMGFIPVMTSVF H NSQEWSWQTI LMGFSFLVFLLVARH VSM RRPKLFWVSAAAPLVCVI LSTFLVFAFKA QHHGFSVIGKLQEGLNPPSWNMLQFHGGHLGLSMKTGLVTGIISLTEGIAVGRTFASL MNYKVDGNKEMMAIGLMNIVGSSTSCYVTTGAFSRSAVNHNAGAKTAASNIVMSITV MVTILFLMPLFQYTPNWLGAIIVSAVVGLIGIPAAYHVWKMDKFDFWMLCAFFGVIFIS VQHGLAIAVAVSIFKILMQITRPKTVLLGKIPGTDIYRDLHHYKESVKIPGFLILSIEAPINF ANSTYLNERVLRWIEEYEAEDPKMHSNSSLRFVILEMSTVSTIDTSGVSFFKELKQTM ENKGVELVLVNLVGEVMEKLQRSNEAGDFMKPDCLFLTVGEAVATLSATIKSQSSND V

[0426] SEQ ID NO: 23 > KJB56717 MGLNSNRVEDFFSHSANTTVTSLKVSTEIPMPPPEEAVEVHNVCLPPKKTTFQKLKHR LSDIFFPDDPLHGFKNQTWCKKLVLGLQSLFPICQWGLHYNLKLFRSDIISGLTIASLAI PQGISYAKLANLPPIVGLYSSFVPPLIYSVLGSSRHLAVGPVSIASLVMGTMLSGSVSS TDDPILYLQLAFTATFFAGVFQASLGLLRLGFLIDFLSNATLVGFMAGAAVIVALQQLKG LLGIVHFTGKMQFVPVMTSVFDHKDEWSWQTIVMGFGFLLFLLTTRQISIKKPKLFWV SAAAPLTSVILSTILVFCIKSKAHGISYIGHLPKGLNPPSLNMLYFNGQYLALTIKTGLITG ILSLTEGIAVGRTFASLQNYQVDGNKEMMAIGLMNMAGSCTSCYVTTGSFSRSAVNY NAGAQTAVSN I VLAAAVLVTLLFLM PLFYYTPN VI LAAI I ITAVIGLI DYRAAYKLWKVDKF DFLACMCSFFGVLFISVPLGLAIAVGVSIFKILLHVTRPNTVVLGNIPRTQIYQSLNRYKE ASRVPSFLILAIESPIYFANSTYLQERMLRWVREEEERINESHESTLKCIIIDMTAVSAID TSGIDMLCELRKILEKRSLQLVLVNPVGSVMEKMHKSKILESFGMTSMYLTVGEAVGDI AASWKPQP

[0427] SEQ ID NO: 24 > KJB13144 MGVTSDRVEYVCRYGGNGTTTSGVNVSIAAEVTMEIHSVCLPPKESTFQKLKHRLSEI FFPDDPLYRFKDQTWCKKLVLGLQFLFPILQWGSHYDLNLFKSDVVSGLTIASLAIPQGM& C PC932446GB

[0428] 61 ISYAQLANLPPIIGLYSSFVPPLMYSLLGSSRHLAVGPVSIASLVMGTMLSEKVSPVQH PTLYLKLAFTATFFAGLFQASLGFLRLGLVIDFLSKATLVGFMAGAAIIVSLQQLKGLLGI VHFTTKMQLIPVLTSVFHHTKEWSWETIVMGFSFLLLLLTTRHISMKKPKLFWISAAAPL TSVILSTLLVFLIKSKVHGISIIGHLQKGLNPTSTNMLYFNGQYLALAIKTGIVTGILSLTEG IAVGRTFASLKNYQVDGNKEMMAIGLMNMASSCTSCYVTTGSFSRSAVNYNAGAQTA VSNIVLAGTVLVTLLFLMPLFYYTPNVILAAIIITAVIGLIDYKAAYKLWNVDKLDFLACICS FFGVLFISVPLGLGIAVGVSVFKILLHVTRPNTLVLGNIPATQIYQSLNRYKEASRVPSF LI LAI ESPIYFANSTYLQERLLR WQEEEERI KAN H ESTLKCI 11 DMTSVTAI DTSGI DM VCELKKMLEKRSLVLVLVNPVGSVMEKMHRSNILGTNSLFLTVGEAVADISASWKPQP

[0429] SEQ ID NO: 25 > KJB15438 MEYSNSSNTEQPQTCLEITMEVHRVVPPPHKSTIHKLKTTLKETFFPDDPLRQFKGQP TGKKWILAAQYIFPILQWGPHYSLGLFKSDIVAGLTIASLAIPQGISYAKLANLPPIVGLY SSFVPPLVYAVLGSSRDLAVGPVSIASLILGSMLRQEVSPVSDPVLFLQLAFTTTFFAG LFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKSLLGITHFTTKMGLVPVLSSVF HNTEEWSWQTILMGFCFLVFLLVARHVSMKRPKLFWVSAGAPLVSVILSTLLVFAFKA QHHGISIIGKLQEGLNPPSWNMLQFRGSHLGLSIKTGLVTGIISLTEGIAVGRTFAALKN YKVDGNKEMMAIGLMNMVGSSTSCYITTGAFSRSAVNHNAGAKSAVSNIVMSITVMV TLLFLMPLFQYTPNWLGAIIVSAVVGLIDIPAACQIWKIDKFDFIVMLCAFFGVIFISVQD GLAMAILLQITRPKTVMLGNIPGTDIFRDLHHYKESMKIPGFLILSIEAPINFANSTYLNER ILRWIEEYEAEDHKKQSSLQFVVLVMSAVSAIDTSGVSIFKELKKTVEKKGAELVLVNPL GEVMEKLQKSDEAGDFMRPDCLFLTVGEAVATLTATIKSQVSNHW

[0430] Coffea canephora

[0431] SEQ ID NO: 26 > CDP00782 MEVHQVVPPPHMSTFNKLKNRFKETFFPDDPLRQFKGKSTKTKWILGAQYIFPILQWG PNYDLKLLKSDIVSGLTIASLAIPQGISYAKLASLPPIVGLYSSFVPPLIYAVLGSSRDLAV GPVSIASLIMGSMLRQEVSPATDPLLFLQLAFSSTFFAGLFQASLGFLRLGFIIDFLSKA TLIGFMAGAAIIVSLQQLKSLLGITNFTNQMGIVPVLSSVFHRTNEWSWQTILMGFSFLA FLLLTRHIGIKKPKLFWVSAGAPLVSVILATLVVFASKAQHHGISVIGKLQEGLNPPSWN MLHFHGSHLGLVMKTGLITGIVSLTEGIAVGRTFAALKNYQVDGNKEMIAIGVMNIVGS STSCYVTTGAFSRSAVNHNAGCKTAASNIIMAVTLMVTLLFLMPLFQYTPNVILGAIIVT AWGLIDIPAAYQTWKVDKFDFIVLLCAFLGVLFISVQGGLAIAVGISIFKVLLQITRPKT VMLGNIPGTDIYRNLHQYKDAVRIPGFLILSIEAPINFANTTYLKERITRWTEDYEAEVE KTKKQSGLRFLVIDLSAVSAIDTSGISFFKELRMVLEKKGIEASQQQLQRM& C PC932446GB

[0432] 62

[0433] SEQ ID NO: 27 > CDO98069 MGINSNRVEHCSDHHACHDEHETAVTISSTDVNVMPSLEVHRVCLPPHKTTLEKLMH KLSEAFFPDDPLHKFKNQTWFNKLVLGLQFFFPIFQWAPNYNFRLLRSDFISGLTIASL AIPQGISYAKLANLPPIIGLYSSFVPPLIYSVLGSSRHLAVGPVSIASLVMGTMLNEAVSY TDEPTLYLQLAFTATFFAGLFQASLGLLRLGFIIDFLSKATLVGFMAGAAVIVSLQQLKG LLGIVHFTSKMQIVPVVASVVQHKHEWSWQTIVLGVSFLILLLTTRNISLRKPKLFWISA ACPLASVILSTILVVLFKSKLGGVQTIGHLTKGLNPPSSNMLQFRGPFLAIAIKTGIVTGIL SLTEGIAVGRTFAALKNYQVDGNKEMMAIGFMNMAGSCSSCYVTTGSFSRSAVNYNA GAQTVVSN VI MAAAVLVTLLFLM PLFYYTPSVI LGAI I ITAVIGLI DYQAAYKLWKVDKLDF LACLCSFLGVLFISVPLGLAIAVGVSVFKILLHVTRPNTVVLGNIPGTQIYQNISRYTEAL RVPSFLVLAVEAPFYFANATYLQERILRWVREEEERIQANNESKLKCIILDMTAVTAIDT SGIDTICEVRKALENRSLKLVLANPVGSVMEKLHQSNILDSFGLDGLYLTVGEAVADIS SSWKP

[0434] Vitis vinifera (grape)

[0435] SEQ ID NO: 28 MGVSSNRVEDFSSHHETSVRMSPASAEAVMVVAMPPVEIHRVCLPPSKTTFQKLRQ RLSEIFFPDDPLHRFKNQSSFTKLVLALQFFFPIFHWAPTYSLALLRSDIISGLTIASLAIP QGISYAKLANLPPIIGLYSSFVPPLIYSILGSSRHLAVGPVSIASLVMGTMLNNAVSCSA DPILYLKLAFTATFFAGLFQAALGLLRLGFIIDFLSKATLVGFMAGAAVIVSLQQLKGLLG IAHFTTKMQIVPVLTSVFQQRHEWSWQTIVMGFGFLAFLLITRQISMRRPKLFWVSAA APLTSVILSTLLVFLLKSKLHGISIIGHLPKGLNPPSSNMLYFHGSYLAVAIKTGIITGILSL TEGIAVGRTFAALRNYQVDGNKEMMAIGFMNMAGSCSSCYVTTGSFSRSAVNYNAG AQTAVSN 11 MASTVLVTLLFLM PLFH YTPN Fl LAAI I ITAVIGLI DYEAAYKLWKVDKLDCFA CLCSFFGVLFISVPLGLAIAVGVSVFKVLLHVTRPNTMVLGNIPGTQIYQNPSRYREAM KVPSFLILAVESPIYFANSTYIQERILRWVREEEEQIQANNGNALKCVILDMTAVTAIDTS GIDVICELRKMLEKRSLQFVLANPAGNVMEKLHQSKILDSFGLNGLYLAVGEAVADISS LWKAQP*

[0436] SEQ ID NO: 29 MEPNASNLHSHCVEITMEVHKVVPPPHRSTFQKFKTRLKETFFPDDPLRQFKGQPPK RKWILGAQYVFPILQWGPNYSLKLFKSDIVSGLTIASLAIPQGISYAKLANLPPIVGLYSS FVPPLVYAALGSSRDLAVGPVSIASLILGSMLRQEVSPSKDPILFLQLAFSSTFFAGVV QASLGILRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKALLGITHFTKQMGLVPVLGSVFHM& C PC932446GB

[0437] 63 NTAEWSWQTIVMGFCFLSLLLLARHVSMKKPNLFWVSAGAPLASVIISTLLVFAFKAQ HHGISIIGKLQEGLNPPSWNMLHFHGSYLGLVMKTGLVTGIISLTEGIAVGRTFAALKG YKVDGNKEMMAIGLMNIVGSSTSCYVTTGAFSRSAVNHNAGAKTAASNIIMAVTVMVT LLFLM PLFQYTPN VVLGAI I VTAVVGLI DLPAAYQI WKI DKFDFI VLLCAFLGVI FISVQQG LAIAVGISIFKVLLQVTRPRTGMLGNIPGTDIYRNIHHYKDGMKVPGFLILSIDASINFANT TYLNERILRWVEEYEAQDAEEEGKKHSSLQFVILDLSAVSTIDTSGVSIFSDLKKALEKK GLEMALVNPVGEVMEKLQRWDEGRDILRPDSVYLTVGEAVASLSSAVKCQPSNRA*

[0438] SEQ ID NO: 30 MGVDSKRVEADSSAVSTPETPFNLTLSIGPVLLQPLEIHRVCLPPQKTTFQKLKHRLSE IFFPDDPFHRFKNQTFLRKWLGLHCLFPILQWVPSYSLSTFRSDLVSGLTIASLAIPQG ISYAKLANLPPIIGLYSSFVPPLIYSLLGSSKHLGVGPVSIASLVMGTMLSETVSSTHESD LYLRLAFTATFFAGLFQASLGLFRLGFIIDFLSKATLVGFMAGAAVIVSLQQLKGLLGIV HFTKKMQIIPVMSSVFGHTKEWSWKTIVLGFGFLIFLLTARLTSLKRPKLFWISAAAPLT SVILSTLLVYLLKSELHGVSVIGELPDGLNPPSANILYFHGPHLGLAIKAGIVTGILSLT EGIAVGRTFASLQNYQVDGNKEMMAIGLMNMVGSCSSCYVTTGSFSRSAVNYNAGA KTAFSNIVMAGAVLVTLLFLMPLFYHTPNLVLAAIIITAVIGLIDYNAAFLLWKLDKLEFLA CLCSFFGVLFISVPMGLAISVGVSVFKILLHVTRPNTVALGNIPGTQIYQNVSRYENASR VPCFLILGIESPIYFANSTYLQERILRWVWEEEERLKEKEENLKCVVLDMTAVTAIDSSG IDAIYELRKTLLNRSVQLVLVNPVGSVMEKLHHSKILDLFGTNQLYLTVGEAVTDISSSW KA*

[0439] Glycine max (soybean)

[0440] SEQ ID NO: 31 KRH50381.1 3;3 orthologue MEVALDKVTMDQVPNEVHQWAPPYKSSLQKFITKVKETFFPDDPLRQFKGQPLKRK LILGAQYVFPVLQWAPSYSFKLFKSDLISGLTIASLAIPQGISYANLANLPAILGLYSSFV PPLVYVVLGSSMDLAVGPVSIASLVLGSMLTEEVSPSEQPDLFLQLALTSTFFAGIFQA ALGILRLGFIIDFLSKAILIGFMAGSAVIVALQQLKGLLGIKHFTKKMALVPVLSSVFQNK HEWSWQTILMGVCFLVFLLVARHISIRKPKLFWVSAGAPLVSVIISTVLSSVIKAQLHGI SVIGKLPQGVNPPSVDKLLFQGSHLGLAIKTGLVTGLLSLTEGIAVARTFASIRNYKVDG NKEMMAIGFMNVVGSTTSCYVTTGSFSRSAINHNAGAKTAMSNLVMSVTVLVTLLFL MPLFQYTPNVILGTIIITAVIGLIDLPSAYLIWKLDKFDFWMLTAFFGVIFISVQLGLAIAV GLSVFRILLQVTRPKTVMLGNIPATTIYRNIHHYNEATRVPGFLILSIEAPINFANITYLNE RILRWVDEEEATINDNLCLQFVILEMSAVSAIDTSGVSLFKDLKTTLTMKGVQLVLVNPL ADVIEKLQKADEVDDFVREDYLFMTVGEAVTSLSSLMKGQSPTMEEEEAQKIVTEY*M& C PC932446GB

[0441] 64

[0442] SEQ ID NO: 32 > KRH44377.1 3;4 orthologue MGVNSYSNSRVEHLACNNNGSNNNMKIQAEIQMPPLEIHKVRLPPERTTLQKLRHRL SEIFFPDDPLHRFKNQTCLMKLLLALQYFFPIFQWAPHYNLSLLRSDIISGLTIASLAIPQ GISYAKFANLPPILGLYSSFVPPLIYSLLGSSRHLGVGPVSIASLVMGSMLSETVSFSQD PILYLKLAFTATFFAGLFQSSLGILRLGFVIDFLSKATLVGFMAGAAIIVSLQQLKGLLGIV HFTNKMQITPVLISVFKQRDEWSWQNLLLGFSFLLFLLTTRHISLKKPKLFWVSAAAPL TSVILSTIFVFILRNKTHKIAIIGELPKGLNPPSSNMLYFNGPYLALAIKTGLVTGILSLT EGIAVGRTFAALKNYQVDGNKEMMAIGLMNIAGSCSSCYVTTGSFSRSAVNYNAGAQ TAVSNIIMASAVLVTLLFLMPLFYYTPNVVLAAIIITAWGLIDYQGAYKLWKVDKLDFLA CLCSFFGVWFISVPLGLGIAVAISVFKILLHVSRPNTLVLGNIPGTPIFHSLNQYREALRI PSFVILAVESPIYFANSTYLQERILRWVREEEERVKANNESTLKCIILDMTAVTAIDTSGI DTLYELRKLVLANPVGNVMEKLHQSNILDSFGLKGVYLSVGEAVADISSSWKAQP*

[0443] SEQ ID NO: 33 > KRH47059.1 3;4 orthologue MGVNSYSNSMKIQAEIQMPPLEIHKVRLPPERTTLQKLRHRLSEIFFPDDPLHRFKNQT CLIKLLLALQYFFPIFQWAPLYNLSLLRSDIISGLTIASLAIPQGISYAKLANLPPILGLY SSFVPPLIYSLLGSSRHLGVGPVSIASLVMGSMLSETVSYSQDPILYLKMAFTATFFAG LFQSSLGILRLGFVIDFLSKATLVGFMAGAAIIVSLQQLKGLLGIVHFTNKMQITPVLISV FKQRDEWSWQNLLLGFSFLLFLLTTRHISLKKPKLFWVSAAAPLTSVILSTIFVFILRNK THKIAIIGGLPKGLNPPSSNMLYFNGPYLALAIKTGLVTGILSLTEGIAVGRTFAALKNY QVDG N KEM M Al G LM N I AGSCSSCYVTTGSFSRSAVN YN AGAQTAVSN 11 M ASAVLVTL LFLMPLFYYTPNVVLAAIIITAVSGLIDYQAAYKLWKVDKLDFLACLCSFFGVLFISVPLG LGIAVAISVFKILLHVSRPNTLVLGNIPGTPIFHNLNQYREALRIPSFIILAVESPIYFAN STYLQERILRWVREEEERVKANNESTLKCIILDMTAVTAIDTSGIDTLCELRKVLEKRSL QLVLTNPVGNVMEKLHQSNILDSFGLKGVYLSVGEAVADISSSWKAQP*

[0444] SEQ ID NO: 34> KRG89334.1 METNNACTMHSHCIEMSMEVHQVVPPPHKSTLQKLKGRLKETFFPDDPLRQFKGQP LKRKLILGAQYVFPILQWGPKYNLKLFKSDLVSGLTIASLAIPQGISYAKLASLPPIVGLY SSFVPPLVYAVLGSSKDLAVGPVSIASLVMGSMLHQEVSPTTDPILFLQLAFTSTLFAG LFQALLGILRLGFIIDFLSKAILIGFMAGAAIIVSLQQLKSLLGITHFTNQMGLIPVMTSVFH NIHEWSWQTILMGICFLVLLLLARHVSIRKPKLFWVSAGAPLMCVIISTLLVFAIKAQNH GISVIGKLQEGINPPSWNMLLFHGSHLDLVMKTGLITGILSLTEGIAVGRTFAALKNYKV DGNKEMMAIGFMNVVGSFTSCYVTTGAFSRSAVNNNAGAKTAVSNVVMSVTVMVTLM& C PC932446GB

[0445] 65 LFLMPLFQYTPNVVLGAIIVTAVIGLIDLPAACNIWKIDKFDFVVMMTAFLGVLFISVQGG LALAVGLSTLKILLQITRPKTVMLGKIPGTDIYRNLDQYKEAVRIPGFLILSIEAPINFANIT YLNERTLRWIEEEEDNIKEQLSLRFLVLEMSAVSAVDTSGISLFKELKATLEKKACVGQ SSC*

[0446] SEQ ID NO: 35 > |KRH09839.1 3;4 orthologue MGVNSNRVEHFDSHESTIKIQDETMQIHAVQLPPHRTTLHKLRQRVSEIFFPDDPLYR FKNQTCFKKFLLALQYLFPIFQWAPNYNLTLLRSDLISGLTISSLAIPQGISYAKLANLPPI IGLYSSFVPPLIYSLLGSSRHLGVGPVSIASLVMGSMLSEKISYTQEPILYLGLAFTATF FAGVFQASLGILRLGFVIDFLSKATLVGFTGGAAVIVSLQQLKGLLGIVHFTSKMQIIPV MISVFKQRHEWSWQTILLGFGFLVFLLTTRHISLRKPKLFWVSAAAPLTSVILSTILVFL LRNTTHQISVIGHLPKGVNPPSANMLYFNGPYLALAIKTGIITGILSLTEGIAVGRTFAS LKN YQVDG N KEM M Al GLM N I AGSCSSCYVTTGSFSRSAVN YN AGAQTTVSN 11 M AAAV LVTLLFLMPLFYYTPNVVLAAIIITAVIGLIDYQSAYKLWKVDKLDFLACLCSFFGVLFISV PLGLGIAVIISVFKILLHVTRPNTLVLGNIPGTQIFHNINQYIEALRVPSFLILAVESPI YFANSTYLQERI LRWVREEEEH I KAN NGAPLKCI I LDMTAVTAI DTSGLDTLCELRKM LE KRSLELVLANPVGNVMEKLHKSNILDSFGLKGVYLTVGEAVADISSIWKAQP*

[0447] SEQ ID NO: 36 > KRH50383.1 - 3;3 orthologue MEPNNACTMHSHCIEMSMEVHQWPPPHKSTLQKLQGRLKETFFPDDPLRQFKGQP LKRKLILGAQYVFPILQWGPKYNLKLFKSDLVSGLTIASLAIPQGISYAKLASLPPIVGLY SSFVPPLVYAVLGSSKDLAVGPVSIASLVMGSMLRQEVSPTADPILFLQLAFTSTLFAG LFQASLGILRLGFIIDFLSKAILIGFMAGAAIIVSLQQLKSLLGITHFTNQMGLIPVMTSVF HNIHEWSWQTILMGICFLVLLLLARHVSIKKPKLFWVSAGAPLMSVIISTLLVFAIKAQNH GISAIGKLQQGINPPSWNMLLFHGSHLGLVMKTGLITGILSLTEGIAVGRTFAALKNYKV DGNKEMMAIGFMNVVGSFTSCYVTTGAFSRSAVNNNAGAKTAVSNVVMSVTVMVTL LFLMPLFQYTPNVVLGAIIVTAVIGLIDLPAACNIWKIDKFDFVVMLTAFLGVLFISVQGG LALAVGLSTFKILLQITRPKTVMLGKIPGTDIYRNLDQYKEAVRIPGFLILSIEAPINFANIT YLNERTLRWIEEEEEDNIKEQLSLRFLVLEMSAVSAVDTSGISLFKELKATLEKKGVELV LVNPLAEVIEKLKKADEANDFIRADNLFLTVGEAVASLSSAMKGQSSTITEGTHTIVSHN

[0448] SEQ ID NO: 37 KRH23482.1 Glycine max - 3;4 orthologue MGVNSNRVEHFASHDSAIEETMQIHAVQLPPHQTTLHKLRHRVSEIFFPDDPLHRFKN QTRFKKFLLALQYLFPIFDWAPNYNLTLLRSDLISGLTIASLAIPQGISYAKLANLPPILGLM& C PC932446GB

[0449] 66 YSSFVPPLIYSLLGSSRHLGVGPVSIASLVMGSMLSDKISYTQEPILYLGLAFTATFFAG VFQASLGILRLGFVIDFLSKATLVGFTGGAAIIVSLQQLKGLLGIVHFTSKMQIIPVTISVF KQRHEWSWQTILLGFGFLVFLLTTRHISLRKPKLFWVSAAAPLTSVILSTILVFLLRNKT HQISVIGHLPKGVNPPSANMLYFNGPYLALAIKTGIITGILSLTEGIAVGRTFASLKNYQV DG N KEM M AIG LM N I AGSCSSCYVTTGSFSRSAVN YN AGAQTTVSN 11 M AAAVLVTLLF LMPLFYYTPNVVLAAIIITAVIGLIDYQSAYKLWKVDKLDFLACLCSFFGVLFISVPLGLGI AVI ISVLKI LLH VTRPNTLVLGN I PGTQI FH N I NQYKKALRVPSFLI LAVESPI YFANSTYLQ ERILRWVREEEEHIKANNGAPLKCIILDMTAVTATDTSGLDTLCELRKMLEKRSLEFVL ANPVGNVMEKLHKSNILDSFGLKGVYLTVGEAVTDISSIWKAQP

[0450] Solanum lycopersicum (tomato)

[0451] SEQ ID NO: 38 > 3;4 orthologue MTLNSIKVEDSSGNTNDGTEASSSSSSSQPNGVHKVCLPPHRTTFQKLRHRLSEIFFP DDPLHKFKNQTALRKFVLGLQFFFPVFEWGPKYNLMLLRSDIIAGITIASLAIPQGISYA KLANLPPIIGLYSSFVPPLIYSVLGSSRHLAVGPVSIASLVMGTMLSQAVTYSKEPTLYL QLAFTSTLIAGCLQAAMGFFRLGFIIDFLSKATLLGFMAGAAVIVSLQQLKGLLGIVHFT N KMAI I PVLTSVFEN RN EWM WQTI MGGCFLI FLLTARQISARN PKLFWVSAAAPLVSV ILSTVIVYLIKNETHVIPTIGHLPKGINPPSVNKLHFGGPYMALALRVGIITGILALTEGIAV GRTFAAMENYQVDGNKEMTAIGLMNIAGSCASCFVTTGSFSRSAVSYNAGGKSVVS NIVMAATVLITLLFLMPLFQYTPNVILAAIIITAVIGLIDYQGAFRLWKVDKLDCIACLSSFF GVLFISVPVGLLIAVGISVFKILLHVTRPNTNALGYISSTRSFQSLSRYTTAVRIPSFLII AVEAPFYFANSTYLHERTLRWIREEEDRIKTNQEPPIKCIIIDMTAVTAIDTSGIDTICE LRRILEKRSLKLVLANPVGNVMEKLFNSNALEAFGLDGLYLTVSEAVDDISSSWKPEK GPAQPLTI*

[0452] SEQ ID NO: 39 > 3;3 orthologue

[0453] Serine151 in bold and underlined - this is the corresponding position to Serine170 in Fang et al., shown to be important for substrate specificity of the transporter.

[0454] TIYLVLVCVVSKKKMEENRVIDITRSSFEVHKVVSPPHRSTLLKLKNRLKETFFPDDPLR QFKGQTIKKKLILGAQYFFPILEWCPNYRFHMFKSDIISGLTIASLAIPQGISYAKLANL PPIIGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLVLGSMLSEVVSPTKDPLLFLQLAFT STFFAGLFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKGLLGIINFTKQMAI IPVLSSVFHTINEWSWQTILMGFCFLVFLLLTRHIGMRKPKLFWVSAGAPLLSVIISTLI VIAIKGQNHGISIIGKLQEGLNPPSWNMLHFSGSYLGLVIKTGIVTGILSLTEGIAVGRTM& C PC932446GB

[0455] 67

[0456] FAALKN YQVDG N KEM I AIG LM N I VGSSTSCYVTTGSFSRSAVN H N AGSKTAVSN I VM A VTVMVTLLFLMPLFQYTPNVVLGAIIVTAVIGLIDIPAAFQIWKIDKFDFLVLLCAFFGVIF VSVQDGLAIAIGISMLKVLMQITRPKTVMLGNIPGTGIYRNVDHYEEALSVAGFLILSIE APINFANVTYLKERISRWIQDYEEEGAKKQPGLRVVVLDLSPVSSIDTSGISLFKDLSM ALEKKGLEFVLVNPIGEVMEKLQRADETKDLMRPDVLFLTVEEAVASLSSTVKYQIPEH

[0457] V*

[0458] SEQ ID NO: 40. SULTR 3;4 ortholog.

[0459] Serine154 in bold and underlined - this is the corresponding position to Serine170 in Fang et al., shown to be important for substrate specificity of the transporter.

[0460] MTLNSIKVEDSSGNTNDGTEASSSSSSSQPNGVHKVCLPPHRTTFQKLRHRLSEIFFP DDPLHKFKNQTALRKFVLGLQFFFPVFEWGPKYNLMLLRSDIIAGITIASLAIPQGISYA KLANLPPIIGLYSSFVPPLIYSVLGSSRHLAVGPVSIASLVMGTMLSQAVTYSKEPTLYL QLAFTSTLIAGCLQAAMGFFRLGFIIDFLSKATLLGFMAGAAVIVSLQQLKGLLGIVHFT N KMAI I PVLTSVFEN RN EWM WQTI VMGGCFLI FLLTARQISARN PKLFWVSAAAPLVSV ILSTVIVYLIKNETHVIPTIGHLPKGINPPSVNKLHFGGPYMALALRVGIITGILALTEGIAV GRTFAAMENYQVDGNKEMTAIGLMNIAGSCASCFVTTGSFSRSAVSYNAGGKSVVS NIVMAATVLITLLFLMPLFQYTPNVILAAIIITAVIGLIDYQGAFRLWKVDKLDCIACLSSFF GVLFISVPVGLLIAVGISVFKILLHVTRPNTNALGYISSTRSFQSLSRYTTAVRIPSFLII AVEAPFYFANSTYLHERTLRWIREEEDRIKTNQEPPIKCIIIDMTAVTAIDTSGIDTICE LRRILEKRSLKLVLANPVGNVMEKLFNSNALEAFGLDGLYLTVSEAVDDISSSWKPEK GPAQPLTI

[0461] Saccharum spontaneum (sugarcane)

[0462] SEQ ID NO: 41 MVGMRGAYGGGGGGGAYNDSKSGRPHGGMAAAAPVTTEQEIAAMAVHKVAPPPA RSTASKM KARVKETFFPDDPFRAFKGQPLGTQWLMAVRYLFPI LDWVPSYSFSLFKS DLGISYAKLASLPPIIGLYSSFVPPMVYAVLGSSRDLAVGPVSIASLIMGSMLRQAVSPT AEPLLFLQLAFTSTFFAGLVQASLGILRLGFVIDFLSKATLVGFMAGAAIIVALQQLKALL GIVHFTTEMGIVPVMASVFHHTSEWSWQTILMGVCFLVFLLSARHVSIRWPKLFWVSA CAPLASVIISTLLVYLFKAQNHGISIIGQLKCGLNRPSWDKLLFDTTYLGLTMKTGLVTGI I SLTEG I AVG RTFASI RGYQVDG N KEM M Al G LM N VVGSCTSCYVTTGAFSRSAVN H NA GCKTAMSNVIMALTVMVTLLFLMPLFVYTPNVVLGAIIIAAVIGLIDLPSVYHIWKMDKM DFLVCVCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPKMMVQGNIKGTDIYRDLHHYKM& C PC932446GB

[0463] 68 EAKEFPAVPAIDTSGIAFLIDIKKSIEKRDLELVLVNPSGEVMEKIQRANEAQNYFRPDC LYLTTAEAVASLSALAKMTKP*

[0464] Solanum tuberosum (potato)

[0465] SEQ ID NO: 42 MEENRVIDITTGSFEVHKWSPPHRSTLLKLKNRLKETFFPDDPLRQFKGQTMKQKLIL GAQYFFPILEWCPNYGFNMFKSDIVSGLTIASLAIPQGISYAKLANLPPIVGLYSSFVPP LVYAVLGSSRDLAVGPVSIASLVLGSMLREVVSPTKDPILFLQLAFSSTFFAGLFQASL GFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKGLLGITNFTKQMAIIPVLSSVFHTINEW SWQTILMGFCFLVFLLLTRHIGMRKPKLFWVSAGAPLLSVIISTLIVFAIKGQNHGISII GKLHQGLNPPSWNMLHFSGSYLGLVIKTGIVTGILSLTEGIAVGRTFAALKNYQVDGN KEM I AIGLM NM VGSTTSSYVTTGSFSRSAVN H NAGSKTAMSN I VMAVTVM VTLLFLM P LFQYTPNWLGAIIVTAVIGLIDIPAAFQIWKIDKFDFLVLLCAFFGVIFISVQDGLAIAIGIS ILKVLMQITRPKTVMLGNIPGTGIYRNVDHYKEAMSVAGFLILSIEAPINFANVTYLKER ISRWIQDYEEEGAKKQSGLRWVLDLSPVSAIDTSGISLFKDLSMALEKKGLEFVLVNPI GEVMEKLQRADETKNLMRPGVLFLTVDEAVGSLSSTVKYQLPEHV*

[0466] SEQ ID NO: 43 MVLSSNRVEDLSTHACNEEGFELPISNHDVPPLEVHRVCLPPHKTTLEKLRQRLLEVF FPDDPLHKFKNQTCLMKLYLGLQFFFPVFEWGPQYNLKLLRPDIISGLTIASLAIPQGIS YAKLANLPPIVGLYSSFVPPLIYSVLGSSRHLAVGPVSIASLVMGTMLSEVVSYTEQPIL YLQLAFTATLFAGVFQASLGFFRLGFIIDFLSKATLVGFMAGAAVIVSLQQLKGLLGMV HFTSKMQIVPVLSSVFQHKDEWSWQTIVMGMCFLAFLLTTRQISTRNPKFFWLSAASP LASVVLSTLVVACLKSKAHGIQTIGHLPKGLNPPSMNMLYLSGPYLPLAIKTGIVSGILAL TEGIAVGRTFAALKNYQVDGNKEMMAIGLMNMAGSCSSCYVTTGSFSRSAVNYNAG AQTVVSNIIMATAVLITLLFLMPLFYYTPIVILAAIIITAVIGLIDYQAALRLWKVDKLDFLAC LCSFFGVLFISVPLGLAIAVGVSVFKILLHVTRPNTGVLGNIPGTQVYQNMNRYRTAVRI PSFLILAVEAPFYFANSTYLQERILRWIREEEERIEANQETAIKCVIIDMTGQLKRKKEEK FADILYLTC*

[0467] SEQ ID NO: 44 MTLNSIKVEDSSGNTNDGAEASSSQSNGVHKVCLPPHRTTFQKLRHRLSEIFFPDDPL HRFKNQTTLRKFVLGLQFFFPVFEWGPKYNLMLLRSDIIAGITIASLAIPQGISYAKLANL PPIIGLYSSFVPPLIYSILGSSRHLAVGPVSIASLVMGTMLSQAVTYSKEPALYLQLAFTA TLIAGCLQAAMGFFRLGFIIDFLSKATLLGFMAGAAVIVSLQQLKGLLGIVHFTNRMEIIM& C PC932446GB

[0468] 69 PVLTSVFENRNEWMWQTWMGGCFLIFLLTARQISARNPKLFWVSAAAPLVSVILSTVI VYLIKNETHAIPTIGHLPKGINPPSVNKLHFGGPFMSLALRVGIITGILALTEGIAVGRTF AAMENYQVDGNKEMIAIGLMNIVGSCASCFVTTGSFSRSAVSYNAGGKSWSNIVMA TTVLITLLFLMPLFHYTPNVILAAIIITAVIGLIDYQGAFRLWKVDKLDCIACLSSFFGVLFI SVAIGLLIAVGISVFKILLHVTRPNTNVLGYISSTRSFQSLSRYTTAVRIPSFLIIAVEA PFYFANSTYLH ERTLRWI REEEDRI KANQEPPI KCVI LDMTAVTAI DTSGI DTICELRRI LEKRSLKLVLANPVGNVMEKLFNSNALEAFGLDGLYLTVSEAVDDISSSWKPEKGPAQ PLTI*

[0469] Beta vulgaris (sugar beet)

[0470] SEQ ID NO: 45 > KMT030423;3 orthologue MKPEEVDHVKMSSISIEQSAISVPNLGIHSVPLPPKKTTLQKLKLKLTEIFFPDDPLYQF KNKSLRTKIILGFQFFFPILQWLPTYSLQLFRSDLVSGLTIASLAIPQGISYAKLANLPP IIGLYSSFVPPIIYSVLGSSRHLGVGPVSIASLVMGAMLSEFISPTDEPVLYLKLAFTST FFAGLFQASLGLFRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKALLGITHFTNKMQIIP VMYSVFSHKNEWSWQTIVMGISFLVFLLTARYISSKKPRLFWISAAAPLTSVILSTLLVF CLRSNGDKIAKIGQLPEGLNPPSINMLYFQTSHIALAAKTGLITGILSLTEGIAVGRTFA SLENYQVDGNKEMMAIGAMNIVGSCSSCYVTTGAFSRSAVNYNAGAKTAFSNIVMAT AVLVTLLFLMPLFYYTPNVVLAAIIITAVIGLIDHEGALRLWKVDKLDFVACLCSFLGVLFI SVPSGLALAVGISVFKVLLHITRPNIVNLGNIPGTQMFNSIERYKNASRIPSFLIIGLEAP IYFLNSTYLQERILRSIREEEDMIKSNSDPMLKYVILDMTAVTAIDTSGIDTINELGKSI QKRSLELALVNPVGSVMEKLHKSKVLDLFGSSNGVYMSVGEAILDMTTSKKVDGLA*

[0471] SEQ ID NO: 46 > KMT007783;4 orthologue MIQLPNNSTTPTEIHRVCLPPKRSTLQKLKHRVSEIFFPDNPLHKFKNQTWLKKLVLSL QFFFPVFLWGPTYNLKLLRSDLISGLTIASLAIPQGISYAKLANLPPIIGLYSSFVPPLIY SILGSSKHLAVGPVSIASLVMGTMLSEWSPTQQPILYLKLAFTSTFFAGVFQAALGFL RLGFVIDFLSKATLIGFMAGAAVIVSLQQLKGLLGMVHFTNQMQIIPVLASVLKYKDEW AWQTILMGFCFLFFLLSARQISAKRPKMFWISAAAPLISVILSTIIVFAVKPTPRQIPIIGY LPKGLNPPSSNMLYFHGSYLALAIKTGIVSGILALTEGIAVGRTFATLNNYQVDGNKEM MAIGLMNMAGSCSSCYVTTGSFSRSAVNYNAGAQTAVSNIVMASAVLITLLFLMPLFY YTPTVILAAIIITAVIGLIDYQAAFRLWKVDKLDFLACVCSFFGVLFVSVPLGLAISVGISV FKILLHVTRPNTTALGNIPGTQIYQSLGRYREAMRIPSFLILSVESPIYFANSTYLQERVL RWVQEEEERVKANSESMMKCVILDMSAVTAIDTSGIELLSELRKILERRSLQLVLANPA GNVMEKLHFSKALESFGSQGLYLTVGEAITDISASWKAQP*M& C PC932446GB

[0472] 70

[0473] Helianthus annuus (sunflower)

[0474] SEQ ID NO: 47 > KAF5758196.1. Helianthus annuus- 3;4 orthologue MGLNSNRVDHYSGSGHAGQVHTFETTTDHPVTITVSTEAMTVHNVCLPPQKSTGQKL RHRLSEVFFPDDPLHGFKNQSRLRKLILALQFFFPVFEWAPNYSLTLLRADVVSGLTIA SLAIPQGISYAKLASLPPIIGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGTMLTEAV PYNQDPVLYLKLAFTATFVAGVFQASLGLLRLGFVIDFLSKATLLGFMAGAAVIVSLQQ LKGLLGIIHFTTKMQIIPVLSSAIEHKNEWSWQTIVMGFCFLAFLLATRHIGMRKPKLFW VSAAAPLASVILSTLLVTLFRSKLHGIATIGHLEKGLNPPSSNMLYFHGEYLGVAIKTGII TGILSLTEGIAVGRTFAALNDYQVDGNKEMIAIGLMNMAGSCSSCYVTTGSFSRSAVN ANAGAKTVVSNIIMASTVLVTLLFLMPLFHYTPNLILAAIIITAVIGLIDYQSAIKLWKVDKL DFLACLSSFLGVLM ISVPIGLAIAVGVSVFKI LLH VTRPNTGVLGN I PGTH IYQN VN RYR EAKRVPSFAI LGI EAPIYFAN ATYLQERIM RWI REEEEWLAAN NGSSLKCVI I DMTAVTG I DTSGLAMM KELKKM LEKRSLQLVLAN PGGTVM EKLHQSN I LESFGLEGVYLTVDEAV TDISSSWKSQP*

[0475] SEQ ID NO: 48 > KAF5793249.1 Helianthus annuus- 3;4 orthologue MGLNSHNNRVDDYSTSVHANEQQNLTTVPVSVTVSGETMAIHSVCLPPEKTTWQKL QHRLSEVFFPDDPLHGFKNQSRVKKLILALQFFFPIFEWAPNYNFTLFRSDVVSGLTIA SLAIPQGISYAKLANLPPIIGLYSSFVPPLIYSVLGSSKHLAVGPVSIASLVMGTMLNDAV PYNKDPNLYLKLAFTATFFAGVFQATLGFLRLGFVIDFLSKATLLGFMGGAAVIVSLQQ LKGLLGITHFTNKMQIYPVLASAIQHRNEWSWQTYVMGFCFLAFLLATRHIGMRKPKL FWVSAAAPLTSVVVSTLLVTLFRSKLHSIATIGHLDKGLNPPSSNMLVFHGQYLALAIKT GIITGILSLTEGIAVGRTFAALNDYQVDGNKEMIAIGLMNMAGSCSSCYVTTGSFSRSA VNANAGAKTVVSNIVMASTVLVTLLFLMPLFRYTPNLILAAIIITAVIGLIDYQSALRLWKV DKLDFVACLSSFFGVLMISVPMGLAIAVGVSVFKILLHVTRPNTCVLGNITGTQIYQNVN RYKEAKRVPYFVILGVEAPIYFANSTYLQERILRWI REEEEWLAAN NGTSVKCVI I DMTA VTGIDTSGLAMVRELKKM LEKRSLQLVLAN PGGSVM EKLHQSN I LESLGSEGVYLTVN EAVADISSAWKA*

[0476] SEQ ID NO: 49 > KAF5762297.1 Helianthus annuus- 3;3 orthologue MEGKFEGDMGSHNHNPCVEIAMEIHKVETPPKRSSLQKISNRLKETFFPDDPLRQFK GQSTKQKWI LAFQYI FPI LQWGPHYN LN LLKSDVVSGLTI ASLAI PQGISYAKLAN LPPI GLYSSFVPPLVYAVLGSSRDLAVGPVSIASLLMGSMLRQEVSPTLDPILYLQLAFTSTF FAGLFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKALLGITHFTKEMGLVPVM SSVFHNTKEWSWQTILMGFCFLVFLLVTRHISIKKPKLFWVSAGAPLLSVVVSTLFVFVM& C PC932446GB

[0477] 71 FKAQHHGISVIGKLEQGLNPPSWNMLHFHGSHIGLVLKTGVITGIISLTEGIAVGRTFAA LKNYQVDGNKEMIAIGAMNWGSTTSCYITTGAFSRSAVNHNAGAKTAVSNIIMAVTV M VTLLFLM PLFQYTPN LVLGAI I VTAVVGLI DLPAAYQI WKVDKFDFVIM LCAFFGVI FIS VQEGLALAVGISIFKMVLQMTRPKTVVLGNIPGTDIFRNKHQYKDAVTVPGFLILSIQAPI SFANSNYLNERILRWIQDYEEEETKSHSDLRFVILDLAAVSAIDANGVMFFQELRRVLE KKGVELVLVNPVGEVMEKLQKADGTHELLQPNNLYLTVGEAVSSLFSSMKVVSC*

[0478] SEQ ID NO: 50 > KAF5757455.1 Helianthus annuus- 3;3 orthologue MERKFEQGLGGSHHHHPCVEIDMEIHKVETPPKRSSFQKIKNRLKETFFPDDPLRQFK GQSSKRKWLLGFQYIFPILQWGPHYNLNLLKSDWSGLTIASLAIPQGISYAKLANLPPI VGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLLMGSMLRQEVSPTQDPLLFLQLAFTS TFFAGLFQASLGFLRLGFIIDFLSKATLIGFMAGAAIIVSLQQLKALLGITHFTKEMGLVP VMSSI FH NTKEWSWQTI LMGFCFLVFLLVTRH ISI KKPKLFWISAGAPLLSVVLSTI 11 FVS KAQHHGISVIGKLEQGLNPPSWNMLHFHGSHIGLVLKTGLVTGIISLTEGIAVGRTFAAL KN YQVDGN KEM I Al G VM N VVGSTTSCYVTTGAFSRSAVN H N AGAKTAVSN 11 M AVTVM VTLLFLMPLFRYTPNLVLGAIIVTAVIGLVDIPAAYQIWKVDKFDFVVMLCAFFGVIFISV QEGLALAVGISIFKMVLQMTRPKTVVLGNIPGTDIFRNKHQYKDAVSIPGFLILSIQAPIS FANSNYLN ERI LRWI LDYEEEEAAKSHSDLRFVI LDLAAVSAI DANGVAFFQELRRVLEK KGVELVLVNPVGEVIEKLQKADGTNDLLQPNNLYLTVGEAVSSLLSSMKVGSSSHG*

[0479] Triticum aestivum (wheat)

[0480] SEQ ID NO: 51 > 3;3 orthologue MVGMSGAYGGHSNGGESRAPKEAVVVRAPKEAVIVGAPPPGPPEMEMAGVVHKVA AQPAQSTASKMKGKVKETFFPDDPFRSFKGQPLRAQWVMAARYLFPVLEWLPGYSL SLFKSDLVAGLTIASLAIPQGISYAKLANLPPIIGLYSSFVPPLVYAVLGSSRDLAVGPVSI ASLIMGSMLRQAVSPSAEPMLFLQLAFTSTFFAGLVQASLGILRLGFIIDFLSKATLVGF MAGAAIIVSLQQLKALLGIVHFTTQMGIVPVMASVFQHTNEWSWQTILMGACFLALLLA ARHVSIRWPRFFWISACAPLASVIVSTLLVFLFKAQNHGISIIGSLKCGLNRPSWDKLLF DPTYLGLTMKTGLVTGIISLTEGVAVGRTFASLKDYQVDGNKEMMAIGLMNIVGSCTS CYVTTGAFSRSAVNHNAGCKTAMSNVVMALTVMVTLLFLMPLFVYTPNVVLGAIIIAAV IGLIDLPAAYNIWKMDKMDFLVCLCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPRMMI QGNIKGTDIYRNLHQYKEAQRVPGFLILTVEAPINFANTNYLNERTKRWIEDESSSGNK QNELRVVILDLSAVPTIDTSGIAFLIDLKKSTEKHGLELVLVNPTGEVMEKIQRANDAHN HFRPDSLYLTTGEAIASLSGFAKMATP*M& C PC932446GB

[0481] 72

[0482] SEQ ID NO: 52 > 3;3 orthologue MVGMSGAYGGHSNGGESRAPNEAVVVRAPKEAVWGAPPSEMEMAGAMGVVHKV AAQPAQSTASKMKGKVKETFFPDDPFRSFKGQPLRAQWVLAARYLFPVLEWLPGYS LSLFKSDLVAGLTIASLAIPQGISYAKLANLPPIIGLYSSFVPPLVYAVLGSSRDLAVGPV SIASLIMGSMLRQAVSPSAEPMLFLQLAFTSTFFAGLVQASLGILRLGFIIDFLSKATLVG FMAGAAIIVSLQQLKALLGIVHFTTQMGIVPVMASVFQHTNEWSWQTMLMGACFLVLL LAARHVSIRWPKFFWISACAPLASVIVSTLLVFLFKAQNHGISIIGSLKCGLNRPSWDKL LFDPTYLGLTMKTGLVTGIISLTEGVAVGRTFASLKDYQVDGNKEMMAIGLMNIVGSCT SCYVTTGAFSRSAVNHNAGCKTAMSNVVMALTVMVTLLFLMPLFVYTPNWLGAIIIAA VIGLIDLPAAYNIWKMDKMDFLVCLCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPRMM IQGNIKGTDIYRNLHQYKEAQRVPGFLILTIEAPINFANTNYLNERTKRWIEDESSSGNK QTELRVIILDLSAVPAIDTSGIAFLIDLKKSIEKHGLELVLVNPTGEVMEKIQRANDAHNH FRPDCLYLTTGEAIASLSGFAKMATP*

[0483] SEQ ID NO: 53 >3;4 orthologue MVGMSGAYGGHSNGGESRAPKAAWVGAPPPEMEMEMAGAMGVVHKVAAQPAQS TASKMKGKVKETFFPDDPFRSFKGQPLRAQWVMAARYLFPVLEWLPGYSLSLFKSDL VAGLTIASLAIPQGISYAKLANLPPIIGLYSSFVPPLVYAVLGSSRDLAVGPVSIASLIMGS MLRQAVSPSAEPMLFLQLAFTSTFFAGLVQASLGILRLGFIIDFLSKATLVGFMAGAAIIV SLQQLKALLGIVHFTTQMGIVPVMASVFQHTNEWSWQTILMGACFLVLLLTARHVSIR WPRFFWISACAPLASVIVSTLLVFLFKAQNHGISIIGSLKCGLNRPSWDKLLFDPTYLGL TMKTGLVTGIISLTEGVAVGRTFASLKDYQVDGNKEMMAIGLMNIVGSCTSCYVTTGA FSRSAVNHNAGCKTAMSNVVMALTVMVTLLFLMPLFVYTPNWLGAIIIAAVIGLIDLPA AYNIWKMDKMDFLVCLCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPRMMIQGNIKGT DIYRNLHQYKEAQRVPGFLILTVEAPINFANTNYLNERTKRWIEDESSSGNKQTELRVV ILDLSAVPAIDTSGIAFLIDLKKSTEKHGLELVLVNPTGEVMEKIQRANDAHDHFRPDCL YLTTGEAIASLSGFAKMATP*

[0484] SEQ ID NO: 54 > 3;3 orthologue MEGTSDRYGGHNGSEILPPKQAVVVKAPQPAVLHKVMAQPAQSSASKMKGKVKETF FPDDPFRSFKGQPLRKKWLMAVKYLFPIVEWLPGYSLSLFRSDLIAGLTIASVAIPQGIS YAKLADLPPIIGLFSSFVPPLVYTMLGSSRDLAVGPTSITSLIMGAMLQKGVPVSPSEEP TLFLQLALTSTLFAGLLQASLGILRLGFIIDFLSKATLLGFMAGAAIIVSLQQLKELLGIIHF TDKMDIVPVMASVFQHTDEWSWQTMLMGACFLVLLLSARHVSMRWPEFFWISACAP LVSIIISTVLIFLFKGENHNISIIGPLKCGLNHPSWDKLLFDPKYLGLTMKTGLVIGIISLTEM& C PC932446GB

[0485] 73 GVAVGRTFASIKDYKVDGNKEMMAIGLMNIVGSCTSCYVTTGGFSRSAVNHNAGCKT AMSNVIMALTVMVTLLFLMPLFVYTPNVVLGAIIIVAVIGLLDFSAAYHIWKMDKIDFLVC FCAFAGVIFLSVQQGLAIAVGISVFRVLMQVTRPRMIIQGNIKGTDIYRNIHQYEEAQRV PGFLILTVEAPINFANSNYLNERTKRWIEDESFSANKQTELRFVIFDLSAVPAIDTSGIAF LIDLKKPTERLGLELVLVNPTGEVMEKIQRANDTHNHFRADCLYLTIGEAIASLSGEAS MATP*

[0486] Sorghum bicolor

[0487] SEQ ID NO: 55 MVGMRGAYGGGGGANNDSKSGRPHHGGGMAGAATTTTEQEIAAMSVHKVAPPPA RSTASKMKARVKETFFPDDPFRAFKGQPLGLQWLMAVRYLFPILDWMPSYSFSLFKS DLVAGLTIASLAIPQGISYAKLASLPPIIGLYSSFVPPMVYAVLGSSRDLAVGPVSIASLI MGSMLRQAVSPTAEPVLFLQLAFTSTLFAGLVQASLGILRLGFVIDFLSKATLVGFMAG AAIIVALQQLKALLGIVHFTTEMGIVPVMASVFHHTNEWSWQTILMGVCFLVFLLSARH VSIRWPKLFWVSACAPLASVIISTLLVYLFKAQNHGISIIGQLKCGLNRPSWDKLLFDTT YLGLTMKTGLITGIISLTEGIAVGRTFASIRGYQVDGNKEMMAIGLMNWGSCTSCYVT TGAFSRSAVNHNAGCKTAMSNVIMALTVMVTLLFLMPLFVYTPNWLGAIIIAAVIGLIDL PAVYHIWKMDKMDFLVCVCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPKMMVQGNIK GTDIYRDLHHYKEAQRVSGFLILAIEAPINFANCNYLNERIKRWIEEESFEQDKHTELHF IILDLSAVPTIDTSGIAFLIDIKKSIEKRGLELVLVNPTGEVMEKIQRANEAQNYFRPDCLY LTTAEAVASLSALAKMTKP*

[0488] Zea mays

[0489] SEQ ID NO: 56 MVGMRGAYGGACNDDSKSRLHGGKAAEPEIASMAVHKVAPPPARSTASKMKVRVK ETFFPDDPFRAFKGQPPGTQWLMAVRYLFPILDWVPSYSLSLFKSDLVAGLTIASLAIP QGISYAKLASLPPIIGLYSSFVPPMVYAVLGSSRDLAVGPVSISSLIMGSMLRQAVSPTA EPTLFLQLAFTSTLFAGLVQASLGILRLGFVIDFLSKATLVGFMAGAAIIVALQQLKALLG I VH FTTEMGI VPVM ASVFH HTSEWSWQTI LMGVCFLVFLLSARH VSI RWPKLFWVSAC APLASVTISTLLVFLFKAQNHGISIIGQLKCGLNRPSWDKLLFDTAYLGLTMKTGLVTGII SLTEGIAVGRTFASLKDYQIDGNKEMMAIGLMNWGSCTSCYVTTGAFSRSAVNHNA GCKTAMSNVIMALTVMVTLLFLMPLFVYTPNVVLGAIIIAAVIGLIDFPAVYHIWKMDKM DFLVCVCAFAGVIFISVQEGLAIAVGISIFRVLMQITRPKMMVQGNIKGTDIYRDLHHYK EAQRVSGFLILAIEAPINFANSNYLNERIKRWIEEESFEQDKHTELHFIILDLSAVPAIDTSM& C PC932446GB

[0490] 74 GIAFLIDIKKSIEKRGLELVLVNPTGEVMEKIQRANEAENYFRPDCLYLTTGEAIASLSAL AKMTKP

[0491] Nucleic Acid Sequences

[0492] Table 1. Genomic Sequences (CDS and promoter) of SULTR proteins.

[0493] Gene name / ID provided for Ensembl Plants (accessible at: https: / / plants.ensembl.org / index.html)

[0494] Gene name / ID (Ensembl Plants) Comment

[0495] AT1G23090.1 Arabidopsis 3;3 orthologue

[0496] (SEQ ID NO: 73)

[0497] AT3G 15990.1 Arabidopsis 3;4 orthologue

[0498] (SEQ ID NO: 74)

[0499] HORVU. MQREX.r3.2HG0204020.1 > HvlMM1A (SULTR 3;3)

[0500] (SEQ ID NO: 71)

[0501] HvIMMIC (SULTR 3;3) HvIMMIC (SULTR 3;3) HQRVU. MQREX.r3.7HG0741010.1. HQRVU. MQREX.r3.7HG0741010.1.

[0502] HQRVU. MQREX.r3.7HG0648840.1 HvIMMIB (SULTR 3;4)

[0503] (SEQ ID NO: 72)

[0504] Psat7g 128440.1 Pisum sativum - 3;3 orthologue

[0505] Psat7g252760.1 Pisum sativum - 3;4 orthologue

[0506] BVRB_8g196040 Beta vulgaris - 3;3 orthologue BVRB_9g220590 Beta vulgaris - 3;4 orthologue

[0507] Bo5g126530.1 Brassica oleracea - 3;4 orthologue

[0508] Bo5g039240.1 Brassica oleracea - 3;3 orthologue

[0509] Bo1g124840.1 Brassica oleracea - 3;4 orthologue

[0510]

[0511] M& C PC932446GB

[0512] 75

[0513] Bo3g068170.1 Brassica oleracea - 3;4 orthologue

[0514] Bo7g056410.1 Brassica oleracea - 3;3 orthologue

[0515] BnaA07g10140D-201 Brassica napus SLILTR 3;3 orthologue BnaA09g30120D-201 Brassica napus SLILTR 3;3 orthologue BnaC05g18450D-201 Brassica napus SLILTR 3;3 orthologue BnaA01g28310D-201 Brassica napus SLILTR 3;4 orthologue BnaA03g34000D-201 Brassica napus SLILTR 3;4 orthologue BnaC05g37350D-201 Brassica napus SLILTR 3;4 orthologue BnaC03g39450D-201 Brassica napus SLILTR 3;4 orthologue BnaA05g23640D-201 Brassica napus SLILTR 3;4 orthologue AVESA.00001 b.r3.2Dg0002194.1 Avena sativa (oat)

[0516] AVESA.00001 b.r3.2Cg0002973.5 Avena sativa (oat)

[0517] Csa_5G289590 Cucumis sativus (cucumber)

[0518] Csa_7G446670 Cucumis sativus (cucumber)

[0519] Vitvi09g00399 Vitis vinifera (grape)

[0520] Vitvi01g00423 Vitis vinifera (grape)

[0521] Vitvil 1g00355 Vitis vinifera (grape)

[0522] GLYMA_15G014000.1 Glycine max - 3;4 orthologue

[0523] GLYMA_13G360000.1 Glycine max - 3;4 orthologue

[0524] GLYMA_20G017100.1 Glycine max - 3;3 orthologue

[0525] GLYMA_07G218800.1 Glycine max - 3;3 orthologue

[0526]

[0527] M& C PC932446GB

[0528] 76

[0529] GLYMA_08G207100 Glycine max - 3;3 orthologue

[0530] GLYMA_07G006500 Glycine max - 3;4 orthologue

[0531] Gohir. A05G128366.1 Gossypium hirsutum- 3;4 orthologue

[0532] Gohir. D05G128800.1 Gossypium hirsutum- 3;3 orthologue

[0533] Gohir. D12G154400 Gossypium hirsutum- 3;4 orthologue

[0534] Gohir. D03G064200 Gossypium hirsutum- 3;4 orthologue

[0535] Gohir. A01G151436 Gossypium hirsutum- 3;3 orthologue

[0536] Gohir. A02G121700 Gossypium hirsutum- 3;3 orthologue

[0537] Gohir. D01G144800 Gossypium hirsutum- 3;3 orthologue

[0538] Sspon.05G0022070-2D Saccharum spontaneum (sugar cane) Solyc03g120250.3.1 Solanum lycopersicum (tomato) - 3;4 orthologue

[0539] Solyc05g007980.3 Solanum lycopersicum (tomato) - 3;3 orthologue

[0540] Solyc09g082550.3 Solanum lycopersicum (tomato) Soltu. DM.05G005020.3 Solanum tuberosum- 3;3 orthologue

[0541] Soltu. DM.03G034860.1 Solanum tuberosum- 3;4 orthologue

[0542] Soltu. DM.12G010700.1 Solanum tuberosum- 3;4 orthologue

[0543] PGSC0003DMG400028635 Solanum tuberosum PGSC0003DMG400018422 Solanum tuberosum PGSC0003DMG400002643 Solanum tuberosum

[0544]

[0545] M& C PC932446GB

[0546] 77

[0547] Vitvi01g00423_t001.1 Vitis vinifera- 3;3 orthologue

[0548] Vitvi09g00399.1 Vitis vinifera- 3;4 orthologue

[0549] Vitvi11g00355.1 Vitis vinifera- 3;3 orthologue

[0550] HanXRQr2_Chr16g0775061.1 Helianthus annuus- 3;3 orthologue

[0551] HanXRQr2_Chr16g0726511 Helianthus annuus- 3;4 orthologue

[0552] HanXRQr2_Chr09g0415321 Helianthus annuus- 3;4 orthologue

[0553] HanXRQr2_Chr17g0826901 Helianthus annuus- 3;3 orthologue

[0554] TraesCS4A02G388000.1 Triticum aestivum- 3;4 orthologue

[0555] TraesCS7D02G084100.1 Triticum aestivum- 3;3 orthologue

[0556] TraesCS7A02G499000.1 Triticum aestivum- 3;3 orthologue

[0557] TraesCS7A02G088700.1 Triticum aestivum- 3;7 orthologue

[0558] TraesCS7D02G486000.1 Triticum aestivum- 3;3 orthologue

[0559] TraesCS2A02G508200.1 Triticum aestivum- 3;3 orthologue

[0560] TraesCS2B02G536100.1 Triticum aestivum- 3;3 orthologue

[0561] TraesCS7B02G406100.1 Triticum aestivum- 3;3 orthologue

[0562] TRITD2Av1G273870.1 Triticum turgidum- 3;3 orthologue

[0563] TRITD7Av1G260730.1 Triticum turgidum- 3;3 orthologue

[0564] TRITD4Av1G234520.1 Triticum turgidum- 3;4 orthologue

[0565]

[0566] M& C PC932446GB

[0567] 78

[0568] TRITD7Av1G025830.1 Triticum turgidum- 3;4 orthologue

[0569] TRITD2Bv1G237240.1 Triticum turgidum- 3;3 orthologue

[0570] TRITD7Bv1G210460.1 Triticum turgidum- 3;3 orthologue

[0571] SECCE4Rv1G0268140.1 Secale cereal - 3;4 orthologue

[0572] SECCE2Rv1G0126690.1 Secale cereal - 3;3 orthologue

[0573] SECCE6Rv1G0440390.1 Secale cereal - 3;4 orthologue

[0574] LOC_Os04g55800.1 Oryza sativa - 3;4 orthologue

[0575] LOC_Os06g05160.1 Oryza sativa - 3;4 orthologue

[0576] Zm00001d000204.1 Zea mays - 3;4 orthologue

[0577] Zm00001d002038.1 Zea mays - 3;3 orthologue

[0578] Zm00001eb068160 Zea mays Sobic.010G033800.1 Sorghum bicolor - 3;4 orthologue

[0579] Sobic.006G244000.1 Sorghum bicolor - 3;3 orthologue

[0580] SORBI_3006G244000 Sorghum bicolor

[0581]

[0582] Barley nucleic acid sequences > HORVU. MOREX.r3.2HG0204020|HORVU. MOREX.r3.2HG0204020.1 Hordeum vulgare- 3;3 orthologue

[0583] SEQ ID NO: 71

[0584] Underlined = exemplar promoter sequence.M& C PC932446GB

[0585] 79 TTTCTAAACAGACAGATATTATTCCATTAAGGAGCATTTTCCACAATGGATATTGTT CCATTAATGAACGCAACAACACTAAGAGGGATCATGTTTCAAATGCACACTTTCTA TACAGACAGATATTACTCCATTAAGGAGCACAATAAATGTGTATGTGTCATTCAATA TGACATGAGAAGTCAGCTATATTGTCGGTGGAGAGCTTCAGCGCTAAACCTCCAT TATTAGCACCGTGAAGGGTTTCCATTGTCTTGTCAGTTGATATCCTTTTGAAGTTTC TTGTAGAAACTGGCCTCAAATGTGCACACTCAAGGAAACTAATTTGAGTAAGATCT GATATTTAATTCAGTGAGCTGTGGGTGGAGTAAAGAACCGAATGACCCATTACGA CAAGTGACTACTCACACATCTTTGGACTGCATGTGATAAACGGAAACCACCAGAC CCCTCGTACGTCTTAAGGCCTTGTTGTACTAAAAGACGCACCGTCAATCAACAAAA AGACACACGTATCGTTCAAGAACATACGGTGCATGAGTGAGTCCCTACAAGCATA TACACGGCATGAGATCACCCAATCGCCATGTCCTGGAGGCGAAAGGAAAGCGGT GAATGTTCTCCATCGCAGGGATTGTGGGTGGCATCAAATCCAGGGATCGCATCCG CGAGGAAAACTACGAATTTATCTCGTGCCTTTGCCGAATTATTGAACGCCCCCCAC CCCACCCCACCCCACCCCACCCTACCACCATTGTGTTGTGTTGTTTCTGTGCTAT GTCACCTTTGATTCTTCTCTCCCTTTCTCAAGTTGCGTCCCGCTTTTCCTCTTTTGT AGCTAGCGACCAGCCCTGCGCCTGGGCGAGATGCCAGCCCAGGCCCATCTCGA CATGTACATATACGGACACGCGCGCCTGCGCCGTGCAGTGGCCTGATCACTTCA GCCTGTAGCTAGCAACACCTCTCTCTCTCTCTCTCTCTCTTGCCACAATAATTCCG CATGGTGGGGATGAGCGGCGCCTACGGCGGCCACCACGGCAATGGCGGCGAGG GCCGGGCGGCTCCCAAGCAGGCCGTGGTAGTCGGTGCGCCACCACCGCCGGCA GCGGCCGAGATGGAGATGGAGATTGGGGTGGTGCACAAGGTGGCGGCGCAGCC GGCGCAGAGCACGGCGAGCAAGATGAAGGGGAAGGTGAAGGAGACCTTCTTCC CCGACGACCCGTTCCGGAGCTTCAAGGGGCAGCCGGTGCGGGCGCAGTGGGTG CTGGCGGCCAAGTACCTGTTCCCCGTGCTGGAGTGGGTGCCCGGCTACTCCCTC TCCCTCTTCAAGTCCGACCTCGTCGCCGGCCTCACCATTGCCAGCCTCGCCATCC CCCAGGCATGTGCATGCCCGACTTCGTCTCTTTTCTCACGCACGTAATAACGCAT CCAAATGCGATTTCTATCTATCTAGCCTGTATTGATTTGGAGGCTAGCCTGTGCGC GATCAATCAGTCTCCGCGTACCAACAAACCCGTGACCAACCGTGCTGATGTGCTT TTTTGTGTTGTTGCTGGCAGGGTATTAGCTACGCCAAGCTTGCCAACCTGCCGCC GATCATAGGCCTATGTACGTACATACATGCTTTCCGTTTCTACAGCGTACTGTACC ATATTTATGTAGTCGTATGTGCAAATAAATAATGAACTGGTATGGATGGATGCATG TTTGCAGATTCGAGCTTCGTGCCGCCGCTGGTGTACGCGGTGCTGGGGAGCTCG AGGGACCTGGCGGTGGGGCCGGTGTCGATCGCGTCGCTGATCATGGGGTCGAT GCTGCGGCAGGCGGTGAGCCCGTCGGCGTCGCCGGCGCTGTTCCTGCAGCTGG CCTTCACGTCCACCTTCTTCGCGGGGCTGGTGCAGGCGTCGCTCGGCATCCTCCM& C PC932446GB

[0586] 80 GCCTGGGATTCATCATCGACTTCCTCTCCAAGGCGACGCTGGTGGGGTTCATGG CCGGCGCCGCCATCATCGTCTCGCTCCAGCAGCTCAAGGCCCTGCTCGGCATCG TCCACTTCACCACCCAGATGGGCATCGTCCCCGTCATGGCCTCCGTCTTCCAACA CACCAACGAGGTCCGTCCGTTCGTCCATGGCGTTCAATTATACACGTGCACACGT ACGGTGCATGCTTGGCTAACATAATGTTTTGTGTGTGCGTGCGTGCAGTGGTCGT GGCAGACGATACTGATGGGGGCATGCTTCCTGGTGCTCCTGCTAGCGGCCAGGC ACGTGGTACGTTACGTTACATACCCTTTTTTTTTTCTTTCTTTCTCTGCATGCTGTC GTGGACCGTTGACCGGCGCCGATTATATATGGCCTGCATGCTGGCCTGCGCGCG AGAAGCACCAAAATGCTGCTGTTTCTGCGCTGCTTGAGGACACTTTTATCTTCTCT TCATCACGTTGTCCGTGTGCCAGTACGCGTCGATCGTGCATCTAGCTGTACGCAT ACGCCGTCGGCAGGGCTGCGCTACCCTCTCACGGCACAAAGTTCTAGCAGCAAC GGGCACGTACGTACGTGCCACCACACACACACACACATGTGGTCGCGATCGTAC ACGTCGATGATGCACGACGCATATATATCTCACACCGTCCATGGGAGGGAGGAC GTACGATAAGATTAAGATAGATGTGTGCAACATGTTTATCTTCATTCAGGTTGGTG ACCAGGGGATGGACGCACACATCAGACAGTGCTTTACCCACAAATTACATAAGCT TTTTGATGATCATTATTATGGAAAAAAGGCTAGTAGTAGTAGTAGCGGTCAACGTC GTGTTGGGAAAAGAGGCTTCGGGACAAAGCCGGGCGGTGACTTGTCACGTTCGG TTCACGACGAGGTGGACAACACTGGTAAAGTGCCAATTCGCAGTGATGAAGGGC GTTCACTGAAACTTTGGACCATTTTCCCCACTCCCAGCAAGCTACTGTGACCTCTA CTTCACTGGACCTGCATTTTCATACCGAACCTTAGGTAGCTGGCTTGAGAAACCAA GCGCCCAAAGGCCGTCCAGTGCAACCACTACCCGATCTATTATGTGATGTAGCTA TATATATATATATAGATGAATACTGGTGTGCCTCACGATGCCATACTAATTTCGTTA CTCGGATTAAATGCACAGAATTCAAGTGCTTGTGCACCGTAAGAAATCCAAACTCT GCTCTGTACTACAGTACTTCTGTATAGCTAAACTTTCTTACTGCAATGTATTGAGG GTTTGTGGCCTCTCCACTGTAACTCTATGAACTTGCAACAAAAACTCCCTTACCCC CGCAAAAAAAGAAAAAAAAAACTTCTTACTGCAGGGATGAGATATCTTTCTGGATG AAACTTAGTTGGGAGCAGTAAAGGTCTTGAGCCATACCTAGTTTTAAATTTAATAG CCTGTGCTGGTGGCAACCCTGTCTAACTCAGTTCATTCTTAACTGCTTTTGCTTGG GTTGCCCAGTTGGTGTATTCTGAAACTGACATAGAATTTTACAATTTTACAATCTAA CCAAATCTGCACTCTTGCTGTATGGCTGAATCCATGGGTGCTGGCTTCAGAAATTA AGCTCCCCCAGGCCATAATTAGTGCAACTGCTACAGCTTGATCTATATATGCTAGC TATATATAGAGCAATAATGCTGCGCCTCTCAAGGCCATACTGATTTTGGCGCATCG ACAAAATGGTCATGACAAAGAAAATTAAGAAACTCACACAATTCAAGTACTTCTGC AACCTAGAAATTCTGAAGTTTGCTCTGTAGTCTGTACTTCTCTAGCTACATCTTCTT TCAGTACTGACCTATTTCCAAATGAAAATCGGCTGGGAATAGTGAAGGACTTGAGM& C PC932446GB

[0587] 81 CACTTCAATGGCCAGTGCTGGCAACAACTCTGTTTCACTCTGCTTCCGCTCTGAAA ATGACAGACAGCGTTCCATTTCCAGAACCTAAAATTAAATCCACTCCTCCTTGAAG TTGTTGAGCCTAGCTACCTCCTGCAGTACTGGTGGCAACCTTGTGTAACTCGGTT CATAGTAGTAGTAAAGTTCTTGAGGCATACTTACCTACTTTGATAGCCGGTGCTGG TGGCGACCCTGTCTATCTCACTTCATTCTTAGCTGTTTTTGCCATGGGTGGTTGGT GTATGCTGAAACTGTCAGAGAAATTCCCATTTTTAGAACCTAAAATCATATCTGTTT CTTCTTTTGCAATCACTGATTCTGACTTCTACACTCGTACCTATCCGTTTGACATTC TGCGTGCATGTGTATGTTCAGAGCATGAGATGGCCGAAGTTCTTCTGGATCTCAG CGTGTGCTCCCTTGGCGTCCGTCATCGTATCCACGCTGCTCGTCTTCCTGTTCAA AGCACAGAACCACGGCATCAGCATCGTAAGAAATCAACTAAAACCGAACAATCATT TCTATGATGTACAACTGTGTTATGCAACATGATGGCAAACAAGCAGTGTCTGCATG TTTTCAACGCAGATTGGATCGCTCAAATGTGGCCTGAACCGCCCCTCATGGGACC AACTGCTCTTCGATACCACATATCTTGGACTCACCATGAAGACCGGCCTCGTCAC CGGAATCATCTCCCTGACGGTATGCTATATATGGCCTCACACCGCAATAAAACATT ACCCTACACTTCAAATTATAATTAAGTAGGTGCCGTGGATGTTTCATAACCAGCGT CACTGGTAATCTGATGTGTGCCCTGAATTTCGCTCTGACAGGAAGGGGTAGCAGT TGGTAGGACGTTCGCATCACTCAAGGACTACCAGGTGGATGGAAACAAAGAGATG ATGGCCATTGGGTTGATGAACATTGTGGGCTCATGTACATCATGCTATGTGACAAC AGGTAACCTACATTTCTGGACTTAGAATAACCTCCAGCATTCTCTTGCATTCCTTGT AAAAAAAAAAATTTGAAGTATCAGTACCAGGGTAGAAAGTCTTTTTTTTCTTTCGTC AAAAAAGAAATATTTTTTAGATTATAGAAGGACAAGCCTCCCGGACTCTGCATCAT ACGATAGACACAACTATAGGTAGAAAGCTTGATGTCTCTCTTATTTCTAACAGGAG CATTCTCTCGCTCTGCTGTGAACCACAATGCTGGCTGCAAGACCGCCATGTCCAA CGTGGTCATGGCACTGACGGTCATGGTCACGCTGCTGTTCCTCATGCCATTGTTC GTATACACGCCGAATGTTGTCCTTGGAGCAATCATCATCGCTGCTGTCATTGGCC TGATCGATCTCCCTGCTGCATACAACATCTGGAAGATGGATAAGATGGATTTCCTC GTGTGCCTTTGCGCATTTGCTGGTGTCATCTTCATCTCGGTCCAGGAAGGCCTCG CGATTGCGGTAACCAACATAATTGTTACTGAGATACGTTGCTGAACTTTCTGCAGA GGAAGCTGCTACTTTAATTGCTTCGTTACTGAAATTTGTCAAATTTGCAGGTCGGT ATATCCATATTTAGGGTATTGATGCAAATCACAAGGCCAAGGATGATGATTCAGGG GAACATTAAGGGAACAGATATTTACCGGAACCTCCATCAGTATAAGGAGGCTCAA AGAGTTCCTGGATTCTTGATACTGACAATTGAGGCTCCTATAAACTTTGCAAACAC CAACTACCTAAATGAAAGGTAGGAAAAAAATTCTAGTAACAGAAATAAATGTTGAA GTCAAGCTGTGGAACATATTACAATTTAATATTTCCATTTGTAGGACTAAAAGATGG ATAGAGGATGAAAGTTTTTCAGGGAATAAACAAAGTGAACTCCGTGTCGTAATCTTM& C PC932446GB

[0588] 82 GGATCTATCAGGTTAGTGACTCTCGAGAGAAACATCGTTCACTCTATAGTTAAGCA TTATTATTATTTAACACAAGCTCACTAGACCATATGTATTTTTCTCAGCTGTCCCTG CAATTGACACAAGCGGCATAGCATTCCTCATCGACCTAAAGAAATCAACAGAGAA GCATGGCCTAGAGGTATACTTCATCATATATATGAAAAGTTAATATCTCTAGTTCAC AAGCTATCAGTTTAAGTTATGTGGCAAATTTACCTTTTTTCTCTCCCTTCGCAGCTT GTACTTGTGAATCCAACCGGAGAGGTTATGGAGAAAATACAACGAGCGAACGATG CACACAATCATTTCAGGCAGGACTGCCTCTATTTGACCACCGGGGAAGCAATCGC TTCACTTTCTGGATTTGCCAAGATGGCAACACCT— AGCGGTTTTGCCGAATTGC CATTTTGATCATCTGATTGCAATGCTAAATTTGCATTGCCTTCTCATCAGTGAATAT GCCTGCAACAAAGCATTTCACAAGTTTGTTTGTAGCTATCTTCAGTGCCCTTCAAG TCTATGTGATGGAAATTAGGTGCTTGAAATATGTACATTCATGGTTCGAATCAACAT GAGTTCTCGCA

[0589] SEQ ID NO: 72 > HORVU. MOREX.r3.7HG0648840|HORVU. MOREX.r3.7HG0648840.1 Hordeum vulgare- 3;4 orthologue

[0590] Underlined = exemplar 5’UTR / promoter sequence.

[0591] AAGCTTTCCAGATATTGATAGTGGTTTTGTCTTTGTACATGTATCCTTTTGGTATCC TACTAGTTCACGTTGGTAGTTGAAGACATTGAAGCATGTTGTTCTAGCTAGAAACA ATCTCTTTCCGTCAGCAAGTGTCTAGGTGGGTACGTTGGTTGTCTTCACGCCCCT AACTTTTTTTTACCACACTGAAAGCAAATATCTGGTGATTCTGGCTGTTTACTCTTG AGATAGAACAACATGCTTCAATCGCTTCAACCTCGCTTCAATCGCTTCAACCTCGA ATGAATCCTGTGACGTAAGCACATCTACAGCCGGGCCCTCAAATCCGTCTCAAAC GATCTGTGCAATCCGTCTAGTCATTGACCGGTCACAAAATATTGATCTAGCCGGAC CCTTCGTATCCATCTCAAACGTCCGGGCTGACCAGCATCACTCATATTCATCTTAA ATATAAGAACGATATGAGGGCTTACAGACGCATCCGAACACGCCTGGTCAGTCCG CCATGCAGAACGCAACCCCACCTTGGACCATATTTTTTTCTTTCTTTATTCATTCTC TACTCTCAATCTTTCTTCTTCACAAATCATATGCAAATGACCGGACATATGAGGGA AAATGTAAAAAATATAGCTGCACGGACGAAAAAGTAAGAGTTTAAAACGGACCAGC CGTGTCCGTGGGCGTTTAATGGACCGGATTTGCACGTTTTAATTGTAGATGCTCTA AGGCGGTTCAATATATCCAAACAAAAGACATAAATGAGTACTACGTATACACACGT TTTTGTACGAGGTATAATTCGATTTAGATTCCAGCACCTCCGTAGGTACAAATTAAT TAATTAACTTACCCGGCCTTAAAACAATATTTACCCCCCAACCCGAACGGCCGCTG ATTTTTCCAGCTCTGACCTCGGTGGCTAATTACAGCATCCAGTAAATAGTAACCGG AGAGGGCCCACCCAAATTAACCCAACCCAGCCCTACCACACCACGCCGGTGGCAM& C PC932446GB

[0592] 83 GAGCCCCCCCTTCCTTCCTTCCTTCCTAATCCGAACCCGCAACGCCGAGGGGGG AAATCCTCGGCTTCTCCCGCGCTTTCGCCTCGCGTCCCGTCCCTATAAAACCCGG CCATCCTCCGACGACCAATCCATCCATCCATCCATCCATCCATCAAGCAGGAGCA GCAACAACACACAGCCACCGTCCTCGGCCCTCTGCTCTGCTCCGCTCCGCTCAG CCATCGACCTCCCCTCCCCTGCGCCGGCGGTTGCAGGGAGAGGGCGTGCAGCG TAGCCATGGTGGTGAACAACAAGGTGGAGACCCTGGCGTTCGACGTGGAGGCGG GGCAGGGACAGGGGCCCCGGCCGAAGGGGGCGGCGGAATCGGGGGCGGCGG GGAGGGTGGTGGAGCTGCACAAGGTGTCGGCGCCGGAGCGCAGGACGACGTGC CGGGCGCTGGGGCAGAGGCTGGCGGAGATCTTCTTCCCCGACGACCCGCTGCA CCAGTTCAAGAACCAGTCGCTCGCCCGGAAGCTGGTGCTCGCGCTGCAGTACTT CTTCCCCATCTTCCACTGGGGTTCCAACTACAGCCTCCGCCTCCTCCGCTCCGAC GCCGTCGCCGGCCTCACCATTGCCAGCCTCGCCATCCCCCAGGTAATCCATCCC TCCGTCCATTGGTGATTGGTCCCGCTGCCTTCAATTCGCCGCCCCATTTTTAGCA CCTTCCTTTTCTGGCCCACGGCGAAGCCACCGGCCGCAAGTTTCCTCCATCGGTC GGCATTAATTTCTTTCCATCTTCTCTTCTCCATGCAGGGCATCAGCTACGCCAAGC TCGCCAACCTGCCCCCAATCATCGGCCTATGTGAGTCTCGCTCTCGCTCGCATGC ACGCCTCGTCGTCTACCCATTCATGGCGAGGAATTAATGGCTGATTGACGAAATT GTTGGCATGCGCGCAGATTCGAGCTTCGTGCCGCCGCTCATCTACGCGCTGCTG GGGAGCTCGCGCGACCTGGCGGTGGGCCCGGTGTCGATCGCGTCGCTGGTGAT GGGGTCCATGCTCCGGGAGGCGGTGGCGCCGGAGCAGCAGCCCATCCTGTACC TGCAGCTGGCCTTCACCGCCACCTTCTTCGCCGGCCTCTTCCAGGCGTCGCTGG GGTTCCTCCGCCTCGGCTTCATCGTCGACTTCCTCTCCAAGGCCACGCTCACCGG CTTCATGGGCGGCGCCGCCGTCATCGTGTCCCTGCAGCAGCTCAAGGGCCTCCT CGGCATCGTCCACTTCACCACCCACATGGGCTTCGTCGACGTCATGGCCTCCGTC GTCCGCCGCCACAGCGAGTGGGAGTGGCAGACCATCGTCATGGGCGTCGCCTTC CTCGCAATCCTCCTCGGCACACGCCAAATCGTAAGCCAGCCACCACCCATGATCT GCCTCTGCCTAGCACATACTCCCATATTTACTTCCAATTTAATTTAGTTAAAAGTAG TATACAATGTTTAGTTCCAATTTAATTTCCCCATATTTAGTTTCAATTCTACTAATAA TACAATTAATTAATTCCAAGATCCTCTTACTTTTAACATAAACCATGTTTAGTTTCAA TTTTATACTAATTAAATCACAGATTGTAACCGGAGAAGGACACATATGCATATGCTT ATGGTTATAGGATACATATGCATCTCCTTTTTTTTTCCTTTTGTTTAAGACAGGTTTA GTCATCATTTGCGAATTAGTCGGCTAATGCTTTTTCTAGAATTATTTGACCACGGG CATGGACAGGTAGCAACTAGGAAAGGTAGAATTTGCATAAAGGGGAGGTGGTGAT TAAACCCACAAAAGTGAGGCAGTACAATGGAGGGACCCAAAGGGTGCTTCCCACT AACAAGGAATTTCACTCCACTCAAATTCAGCTAATGGCTAATAATTAGTGGCAGGGM& C PC932446GB

[0593] 84 TGATCTTAACCACCTAGCTCCTGCTTGTTGGGACCTGATAATGCTGGGATATTTAT CTTTCTGAAAGAAAAGAAAGAAAAAAGCATTCCTATTTCAATTTGACTTAACAAATA AACGAATCCATTGCTTTACTTTAGTTTTTTGTTTCCTCGGAACAACATTCGACGGTA TCTGAAGTACACTCAAAATGAGTATCATACATAAGAAAGAAATGTCCTATAATTCAT TAGAGAACATGGCTAATAGCTTGGTCCAATCAGTTACAACTTTTTGAACTCTTTTAG TTTGTCTATGCTAATAGTACTTTAGACGGTCCTAACAAGAGCTTTTTAGCCTTTCTT CGTGTAACTGGGCAGTTGGTAGTCTATTTCCTCCCCCTATGGGAGGCCTAACCTT CCAAGATGTCTGAATAACAATAGACTGAAACATGGTGCCAAAAAGTGCCTGTTGGT TCTTGAGCGCATGGGCTTTCAAGATTGTAGAATGATTGCTAAAGGTGGTTCAAATC AGTATAGTCTTCTCCATTCAAACAAATCAGTGCTACCTATGTCCTTTTCTCCTTCAC AAAAGTGTCACTGTTTCCTTCAGCAAAAAAAAGGTACAGATTAGAAGATTCAGTCA TACCAAACATATGTTTTGAAGCCAAACCTAACCTGTCCGAGCAGTTTCAGTTTGGC CATGCACATACAGCTGCCATGTTCATAGCTTGCGTCAGTTTTCATACACTCTGCTA GGCCAAACGTATCAATCAATTTGGAGTAAGGCGTGTGTCATTGTGTGTGGCCTTA GTTAGAATCTATTCTGTATTGGCCGGTGACCCATTGCAGCCTGCCCTCTTTTCCCT TTTTGGGGAGAATGTGTGGAATTTGGTCTCTGATGTGTCACCTCCTAAGCTAGTAG TTGGTCACAATCTTTCCTCATCTCCATGATATTTTCTTCTGTCAAATCAAATGTGGA GAATCAAATGTGGAGATGATGATTGATGAACAACGCGCCGCGAACCCGCAATAAC GTGTCGCAAGATCCTCGATGACAAGCGGGCAAGGGTGCGTGATGGACTGATGGT TTTCCGATTTCAGAAGTGTTGGTCTTGAAAAGATGGGGCAACATTGACTAAACAGC TTAATTTCCACCTCAGAATTTCCCATATCATAGAAGAACTACTAGTAGTAAACTTAT TTATGGGGCACGCAGTTATCCTTGTCATGTTAAAATCTTCAGTTAGCCTGTCCTCG ATTTGTTTTCCCCTGCTCTGTTTTTTTTGTGTGACGATAATTCAGGTATAGCAATAG CAAGCAGATGTACTGAACAATCACACATTTTTACTCCTTGTTCTTCCGCAGAGCGC TCGGAATCCAAGGCTGTTCTGGGTGTCAGCGGCGGCTCCCCTGACGTCGGTGAT CGCCTCCACCATCATCTCCTACTTGTGCAGAGGCCATGCCATCAGCATCGTAAGC ACATCGATTTTTCTTCAGTGGCATGTCATCTCTGAACATGTTTCGAGCATGAAAAAT CCAAGGAACATTCATAACCTGTCTCTGATCAGTCTCCAAAAATGCCATCCTTGTCA TGCAGATCGGCGATCTCCCCAGGGGAGTGAACCCTCCATCCATGAACATGCTCGT CTTCAGCGGCTCCTACGTCGCCCTGGCCATCAAGACCGGGATCATGACCGGCAT CCTGTCCCTCACCGTAAGATCGATCAATCAGTCTGTGGTTTTTGTTAATCTGAAGA AACTTTTGTTGCGGCATCTTGATCTGATGGTGGATTTTGTTCTTGAATGTTGCAGG AGGGGATCGCGGTGGGACGGACGTTCGCGTCGATCAACAACTACCAGGTGGACG GGAACAAGGAGATGATGGCGATCGGGGTGATGAACATGGCCGGCTCCTGCGCCT CCTGCTACGTCACCACGGGCTCCTTCTCCCGCTCCGCCGTCAACTACAGCGCCGM& C PC932446GB

[0594] 85 GCTGCCGGACGGCGGTGTCCAACATCGTGATGGCCGCGGCGGTGCTGGTCACG CTGCTCTTCCTGATGCCGCTCTTCCACTACACGCCCAACGTGATCCTGTCCGCCA TCATCATCACGGCCGTGGCGGGGCTCATCGACGTCCGCGGCGCGGCCAAGCTG TGGAAGGTGGACAAGCTGGACTTCTGCGCGTGCGTGGCGGCCTTCCTTGGCGTG CTCCTCGTGTCCGTCCAGGTCGGGCTGGCCGTGGCCGTCGGCATCTCGCTGTTC AAGATCCTGCTGCAGGTGACCCGGCCCAACACCGTGGTCATGGGGCTGGTCCCC GGCACGCAGAGCTACCGCAGCATGGCGCAGTACCGCGAGGCCGTGCGCGTGCC GCCGTTCCTCGTCGTCGGCGTCGAGTCGGCCATCTACTTCGCCAACTCCACCTAC CTGGTGGAGAGGATCATGCGGTATCTCCGCGAGGAGGAGGAGCGCGCCGCCAA GGCCAACCTCTGCGGCGTCCGCTGCATCGTGCTCGACATGAGCGGTGAGTCTGT CTGTCTGTCCGAGTGTCTGTCAGCGCAGCGCAAACTAAATCAATCTTCCCGGGCG CGCGCGCGGGGATCGCCATTAATTGTTCTTCGCGCGCGTCCAAGTTGGAAGAGA TTGATTCGGAGCTAGTCTGACGAGCTTGATTGATTGCAGCCGTGACGGCGATCGA CACGAGCGGGCTGGACGCGCTGGCGGAGATGAAGCGGGTGCTGGACAAGCGGG GCATCGACCTGGTGCTGGCGAACCCGGTGGGGTCGGTGACGGAGAGGATGTAC AACTCGGTGGTGGGCGACACGTTCGGGTCGGGCCGCATCTTCTTCAGCGTCGAC GAGGCCGTCGCGGCGGCGCCGTACAAGGCGCAGCCCWiTGCCATAATATGTAT ACCAGCAGTAGGTCGGGTGCATAGAAGAAGAAGAAGGAGATTAAGAAGAAGAAG AAGGAGGATTAATCAGGATGCAGAGTGTTGGTGTGGTTGATTACGACTAGGGAGA TCCATGGAGGGGTTAGGGTAATTAATTAAGGCCGGCGGAATAACGTACGGCGGT GCTGATATCTCTCCTCTCTTCGCTCCCGTGGAGAAGAGGTTTTTGTTTTTTTGATG AAGAAAATGTACCCCTGATCAGCATGCGCGCGCTTGTTGTTGTTGTAGACGAAGA AGAAGAAGAAGGAGGAGGTGTAAGTTGATGCAGATGTAATCAAATGGAAGTTGGT TAATTTCGTAGTCCATCATGTCCAACTCACTCTTACATACCGCTTCTTGGGTCGGA AAGGGACGGTAAATCTACCCAAATGATAAGTGCCACACGTGCCATAATATGTATAC CAGCAGTAGGTCGGGTGCATAGAAGAAGAAGAAGGAGATTAAGAAGAAGAAGAA GGAGGATTAATCAGGATGCAGAGTGTTGGTGTGGTTGATTACGACTAGGGAGATC CATGGAGGGGTTAGGGTAATTAATTAAGGCCGGCGGAATAACGTACGGCGGTGC TGATATCTCTCCTCTCTTCG

[0595] Arabidopsis nucleic acid sequences

[0596] SEQ ID NO: 73. CDS sequence of gene > AT1G23090.4, ATGCTAAGGCAACAAGTATCTCCCGTAGACGATCCTGTTCTCTTTCTACAGCTAGC CTTCTCTTCTACCTTCTTTGCTGGTCTCTTTCAAGCCTCTCTTGGAATCCTCAGGCTM& C PC932446GB

[0597] 86 GGGATTTATAATAGACTTTCTATCAAAAGCGACGCTAATAGGGTTTATGGGTGGAG CAGCCATAATTGTATCACTCCAACAGCTAAAGGGTCTGCTTGGGATAACTCATTTC ACAAAGCATATGAGTGTAGTCCCTGTTCTCTCCTCTGTTTTCCAACACACCAACGA GTGGTCATGGCAAACAATTGTGATGGGAGTTTGCTTCTTGCTCTTCTTGCTCTCGA CACGTCACCTCAGCATGAAGAAGCCGAAGCTGTTTTGGGTCTCAGCCGGAGCAC CACTTCTTTCCGTTATCGTCTCTACACTTCTTGTCTTTGTTTTCAGAGCCGAGCGTC ACGGAATCAGCGTCATCGGGAAATTACCAGAAGGTTTGAATCCACCGTCTTGGAA CATGCTTCAGTTTCACGGTAGTCATCTCGCACTCGTCGCCAAAACCGGACTCGTC ACCGGAATCGTCTCCCTCACGGAAGGAATCGCAGTGGGAAGAACATTTGCAGCG CTAAAGAACTACCACGTAGATGGAAACAAAGAGATGATCGCCATTGGTCTGATGA ACGTAGTAGGCTCTGCCACATCTTGCTACGTCACAACCGGAGCATTCTCTAGATC AGCGGTCAACAACAACGCGGGAGCTAAAACCGCAGTTTCAAACATTGTTATGTCG GTCACTGTTATGGTTACGCTTCTCTTCCTAATGCCGCTTTTCGAATACACTCCCAA TGTGGTCCTCGGTGCCATCATTGTGACCGCGGTCATTGGTCTCATCGACCTTCCC GCGGCCTGTCACATATGGAAGATCGATAAATTTGATTTCTTGGTGATGCTTTGCGC GTTCTTTGGTGTCATTTTCCTATCCGTTCAAAACGGTCTAGCCATTGCGGTGGGGC TATCGTTGTTCAAGATATTGATGCAAGTAACAAGGCCGAAAATGGTTATAATGGGT AATATTCCTGGAACGGATATATACCGAGATCTTCATCATTACAAAGAAGCACAAAG GATCCCGGGATTTCTTGTTTTAAGCATCGAATCTCCTGTCAATTTCGCCAATTCTAA CTACCTCACTGAAAGAACATCTCGTTGGATTGAAGAATGCGAAGAAGAGGAAGCT CAAGAGAAGCATTCTAGCCTACAGTTCTTGATTCTTGAAATGTCAGCCGTGAGCG GTGTAGACACAAACGGAGTGTCCTTTTTTAAGGAACTTAAGAAAACAACCGCCAAG AAGGACATCGAGCTTGTGTTTGTGAACCCTCTAAGCGAAGTGGTGGAGAAGCTTC AAAGAGCTGACGAACAAAAAGAGTTCATGAGGCCCGAGTTTCTCTTCTTAACCGT CGCTGAGGCCGTTGCGTCGCTCTCTCTTAAAGGGCCATCTCTCAGTAACGTTTAA

[0598] SEQ ID NO: 74. CDS sequence of gene > AT1G23090.4, Accession number: NP_173722.1 ATGGGTCATGGCACTAACAGAGTAGAAGACATGGCGTCACCAAACAATGGAACCG CAGGAGAAACGGTCGTGGAGATACACAGCGTTTGTCTACCGCCGAAGAAAACAG CGTTTCAGAAACTGAAAAAACGCGTTGGGGACGTGTTCTTCCCTGATGATCCGTT ACAGAGGTTTAGGAACCAAACATGGAGAAACAGAGTGATTCTAGGTCTTCAAAGC TTGTTCCCGATATTCACATGGGGTTCTCAGTACGATCTCAAGCTTCTTAGGTCCGA TGTTATCTCTGGTCTCACCATTGCCAGTCTCGCCATTCCTCAGGGAATCAGCTATG CAAAGCTAGCAAACTTACCACCTATAGTTGGTCTTTATTCAAGCTTTGTGCCACCAM& C PC932446GB

[0599] 87 CTGATATATGCAGTTCTTGGGAGTTCAAGACATCTTGCAGTTGGTCCTGTCTCAAT AGCATCACTTGTGATGGGCTCAATGCTAAGTGAAAGTGTTTCACCAACACAAGATT CAATTCTATATCTCAAATTGGCTTTCACTTCAACTTTCTTTGCTGGTGTCTTCCAAG CCTCCCTTGGCCTTCTTAGGCTAGGATTTATGATTGACTTTTTGTCAAAGGCAACT TTGATTGGTTTCACTGCTGGTGCTGCAGTTATCGTGTCACTTCAACAACTTAAGGG TCTTCTTGGAATTGTTCATTTCACTGGCAAAATGCAAATTGTTCCTGTCATGTCTTC TGTTTTCAACCATAGATCTGAATGGTCTTGGGAAACTATTGTCATGGGGATTGGC TTCTTGAGCATTCTCTTAACCACAAGACACATTAGCATGAGGAAGCCAAAACTGTT CTGGATATCTGCTGCATCACCTTTGGCATCGGTTATTATATCAACTCTGCTTGTATA CCTCATTAGATCCAAGACTCATGCCATCTCTTTCATTGGACATCTACCAAAGGGTT TGAATCCACCTTCATTAAACATGTTGTACTTCAGTGGTGCTCATCTTGCTCTTGCC ATCAAAACCGGTATCATCACCGGGATTCTTTCTCTTACAGAAGGGATTGCTGTAGG GAGAACCTTTGCATCTCTAAAGAACTATCAAGTTAATGGAAACAAAGAAATGATGG CTATAGGTTTTATGAACATGGCTGGTTCTTGCACCTCTTGCTATGTTACAACAGGG TCTTTCTCTCGATCTGCGGTAAACTATAATGCTGGAGCGAAAACAGCGGTTTCTAA CATTGTGATGGCCTCTGCAGTTCTAGTGACCCTTCTGTTTTTGATGCCACTCTTCT ATTACACTCCTAATGTGATCCTAGCTGCTATCATCTTAACCGCGGTGATAGGCCTC ATTGATTATCAAGCGGCTTACAAGCTTTGGAAAGTTGACAAATTCGATTTCTTCACT TGCTTGTGTTCCTTCTTTGGCGTTCTCTTTGTTTCTGTACCTCTTGGCCTAGCAATA GCAGTGGCAGTTTCAGTTATCAAGATCTTGTTGCACGTAACCAGGCCAAACACTTC AGAATTTGGGAATATCCCAGGGACTCAGATTTATCAGAGTCTTGGAAGATACAGA GAAGCTTCAAGAATCCCAGGCTTCCTAATCCTTGCTATTGAATCTCCTATATAT TTCGCCAATAGCACTTATCTTCAAGACAGGATTTTGAGATGGGCCAGGGAAGAGG AAAATCGGATAAAGGAGAACAATGGTACTACCTTGAAATGCATAATTCTTGACATG ACAGCTGTGTCTGCGATAGACACAAGCGGGCTTGAAGCGGTGTTTGAACTTAGGA GGAGACTCGAGAAGCAATCTCTTCAGCTTGTGCTGGTGAATCCTGTGGGGACTGT GATGGAAAAGCTACACAAGTCCAAGATCATTGAGGCATTGGGTCTGAGTGGACTT TATCTAACAGTTGGTGAAGCTGTGGCTGATCTCTCATCAACATGGAAAGCTAATGG CCAGCCTTGA

[0600] SEQ ID NO: 76. Intentionally skipped sequence.

[0601] SEQ ID NO: 77. Figure 17 Slimm1A-1

[0602] AGTTGTGTCACCACCCCATAGAAGTACTTTGTTGAAACTM& C PC932446GB

[0603] 88

[0604] SEQ ID NO: 78. Figure 17 Slimm1A-1 AGTTGTGTCACCACCCATAGAAGTACTTTGTTGAAACT

[0605] SEQ ID NO: 79 - Figure 17 Slimm1b-1 GTCTTCGAACCTAACGGTGTACATAAGGTATGTTTGCC

[0606] SEQ ID NO: 80 - Figure 17 Slimm1b-1 GTCTTCGCAACCTAACGGTATAAGGTATGTTTGCC

[0607] SEQ ID NO: 81 Figure 14A CCTCTCCCTCTTCAAGTCCGACCTCGTCGCCGGCCTCACCATTGCCAGCCTCGCC ATCCCCCAGGCATGTGCATGCC

[0608] SEQ ID NO: 82. Figure 14A CCTCTCCCTCTTCAAGTCCGACCTCGTCGCCGGCCaTCACCATTGCCAGCCTCGC CATCCCCCAGGCATGTGCATGCC

[0609] SEQ ID NO: 83. Figure 14B AACTACAGCCTCCGCCTCCTCCGCTCCGACGCCGTCGCCGGCCTCACCATTGCC AGCCTCGCCATCCCCCAGGTAAT

[0610] SEQ ID NO: 84. Figure 14B AACTACAGCCTCCGCCTCCTCCGCTCCGACGCCGTCGCCGGCCaTCACCATTGC CAGCCTCGCCATCCCCCAGGTAAT

[0611] SEQ ID NO: 85. Figure 17 Slimm1B-2 CACAAGTTCAAGAACCAAACGGCGTTGAGGAAATTTGT

[0612] SEQ ID NO: 86. Figure 17 Slimm1B-2 CACAAGTTCAAGAACCCAAACGGCTTGAGGAAATTTCGT

[0613] SEQ ID NO: 87. sgRNA in Figure 14. GAGGCTGGCAATGGTGAGGC

Claims

M& C PC932446GB89CLAIMS:

1. A genetically altered plant, plant part thereof or plant cell, wherein said plant is characterised by reduced expression and / or activity of a sulfate transporter (SULTR) subfamily 3;3 and sulfate transporter (SULTR) subfamily 3;4, wherein the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56, or a functional variant or homolog thereof, and the SULTR3;4 comprises an amino acid sequence as defined in any of SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 or a functional variant or homolog thereof, wherein where the sulfate transporter is expressed as more than one homeologue in the plant, part thereof or plant cell, all homeologues are mutated; wherein said plant is not Arabidopsis.

2. The genetically altered plant, plant part thereof or plant cell of claim 1, wherein said plant, plant part or plant cell comprises at least one mutation in at least one gene encoding a SULTR3;3 protein and / or at least one mutation in a SULTR3 promoter, and / or at least one mutation in at least one gene encoding a SULTR3;4 protein and / or at least one mutation in a SULTR3;4 promoter, wherein said mutation results in reduced expression and / or activity of the sulfate transporter compared to a wild-type or control plant.

3. The genetically altered plant, plant part thereof or plant cell of claim 2, wherein the mutation is a loss of function mutation or a partial loss of function mutation.

4. The genetically altered plant, plant part thereof or plant cell of claim 1, wherein said plant, plant part or plant cell comprises at least one RNAi construct, wherein the RNAi construct reduces the expression of the SULTR3;3 and SULTR3;4 protein compared to a wild-type or control plant.

5. The genetically altered plant, plant part thereof or plant cell of any preceding claim, wherein said plant has increased fungal disease resistance compared to a wild-type or control plant.

6. The genetically altered plant, plant part thereof or plant cell of claim 5, wherein said plant has increased resistance to at least one of Podosphaera spp, Leveillula spp, Sphaerotheca spp., Blumeria spp, Erysiphe spp, Golovinomyces orontii, Colletotrichum higginsianum, Botrytis cinerea, or Blumeria spp.

7. The genetically altered plant, plant part thereof or plant cell of any preceding claim wherein said plant is a monocot or a dicot, wherein preferably the plant is a crop plant.M& C PC932446GB908. The genetically altered plant, plant part thereof or plant cell of claim 7, wherein the plant is selected from wheat, maize, rice, barley, rye, oat millet, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, carrot, parsnip, turnip, zucchini, pumpkin, cucumber, tomato, watermelon, melon, zucchini, squash, pumpkin, gourd, pea, faba bean, common bean, soybean, chickpea, soybean, pea, sugarbeet, grape, sunflower, rye, blueberry, sugar cane and ryegrass.

9. The genetically altered plant, plant part thereof or plant cell of claim 7, wherein the plant is selected from oak, maple, dogwood, magnolia, catalpa and apple.

10. The genetically altered plant, plant part thereof or plant cell of any preceding claim, wherein plant growth or yield is not affected by the genetic alteration.

11. The genetically altered plant, plant part thereof or plant cell of any preceding claim, wherein said plant is characterised by reduced phytic acid levels compared to a wild-type or control plant.

12. The genetically altered plant part of any preceding claim, wherein the plant part is a seed or grain.

13. Use of the genetically altered plant or plant part thereof of any preceding claim as a food or feedstuff.

14. A method of increasing disease resistance in a plant, plant part thereof or plant cell wherein said method comprises reducing the expression and / or activity of a sulfate transporter (SULTR) subfamily 3;3 and / or sulfate transporter (SULTR) subfamily 3;4, wherein the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56 or a functional variant or homolog thereof, and the SULTR3;4 comprises an amino acid sequence as defined in any of SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 or a functional variant or homolog thereof.

15. A method of producing a plant, plant part or plant cell with increased resistance to disease, the method comprising reducing the expression and / or activity of a sulfate transporter (SULTR) subfamily 3;3 and / or sulfate transporter (SULTR) subfamily 3;4, wherein the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54- 56, or a functional variant or homolog thereof, and the SULTR3;4 comprises an amino acid sequence as defined in any of SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 or a functional variant or homolog thereof.M& C PC932446GB9116. The method of claims 14 or 15, wherein the method comprises increasing resistance to at least one fungal disease and / or an oomycete pathogen.

17. The method of claim 16, wherein resistance to at least one fungal disease is selected from resistance to at least one of Podosphaera spp, Leveillula spp, Sphaerotheca spp., Blumeria spp, Erysiphe spp, Golovinomyces orontii, Colletotrichum higginsianum, Botrytis cinerea, or Blumeria spp.

18. The method of any of claims 14 to 17, wherein said method comprises introducing at least one mutation into at least one gene encoding a SULTR3;3 protein and / or introducing at least one mutation into a SULTR3;3 promoter, wherein said mutation results in reduced expression and / or activity of SULTR3;3 compared to a wild-type or control plant.

19. The method of any of claims 14 to 18, wherein the method comprises introducing at least one mutation into at least one gene encoding a SULTR3;4 protein and / or introducing at least one mutation into a SULTR3;4 promoter, wherein said mutation results in reduced expression and / or activity of the SULTR3;4 compared to a wild-type or control plant.

20. The method of any one of claims 14 to 19, wherein where the sulfate transporter is expressed in the as more than one homeologue in plant, plant part or plant cell, all homeologues are mutated.

21. The method of claim 18 or 20, wherein the mutation is a loss of function mutation or a partial loss of function mutation.

22. The method of any of claims 14 to 17, wherein the method comprises introducing and expressing in the plant at least one RNAi construct, wherein the RNAi construct reduces the expression of the SULTR3;3 and / or the SULTR3;4 protein.

23. The method of any of claims 14 to 22, wherein said plant is a monocot or a dicot, wherein preferably the plant is a crop plant.

24. The method of claim 23, wherein the plant is selected from wheat, maize, rice, barley, rye, oat millet, corn, sorghum, oil seed rape, brassica, potato, sunflower, tomato, cotton, carrot, parsnip, turnip, zucchini, pumpkin, cucumber, tomato, watermelon, melon, zucchini, squash, pumpkin, gourd, pea, faba bean, common bean, soybean, chickpea, soybean, pea, sugarbeet, grape, sunflower, rye, blueberry, sugar cane and ryegrass.

25. The method of claim 23, wherein the plant is selected from oak, maple, dogwood, magnolia, catalpa and apple.M& C PC932446GB9226. The method of any of claims 14 to 25, wherein the method does not affect plant growth or yield compared to a wild-type or control plant.

27. The method of any one of claims 14 to 25, wherein the method further reduces phytic acid levels compared to a wild-type or control plant.

28. A method of screening a population of plants and identifying and / or selecting a plant that will have increased disease resistance compared to a control or wildtype plant, the method comprising detecting at least one polymorphism or mutation in a SULTR3;3 and / or 3;4 gene and / or promoter and selecting said plant, wherein the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NOs: 1, 3, 4, 6, 8, 9, 10, 16-31, 36, 39-45, 49-52, 54-56, or a functional variant or homolog thereof, and the SULTR3;4 comprises an amino acid sequence as defined in any of SEQ ID NOs: 2, 5, 7, 11-30, 32-35, 37, 38, 40-44, 46-48, 53, 55, 56 or a functional variant or homolog thereof.

29. The method of claim 28, wherein the polymorphism or mutation is a loss or partial loss of function mutation.

30. A genetically altered plant, plant part thereof or plant cell, wherein said plant is characterised by reduced expression and / or activity of a sulfate transporter (SLILTR) subfamily 3;3, wherein the SULTR3;3 comprises an amino acid sequence as defined in any of SEQ ID NO: 39 or 40 or a functional variant or homolog thereof, wherein the plant is tomato.

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