Enhancing Crop Disease Resistance through Downregulation of Repressor Genes

Abstract

The present invention relates to plants having an increased resistance or tolerance to pathogens. Such plants have a reduced expression level of CPL1 and / or ERF922. The present invention also relates to methods for producing such plants and methods for identifying such plants.

Description

Field of the Invention

[0001] The present invention relates to plants having increased resistance or tolerance to pathogens. The present invention also relates to methods for producing such plants, and methods for identifying such plants. Background of the Invention

[0003] Plants possess an efficient two-tier innate immune system that enables them to resist most microbial pathogens (Jones and Dangl, 2006, “The plant immune system”, Nature, 444(7117):323-329).

[0004] The first layer of defense relies on the recognition of evolutionarily conserved pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) by so-called pattern recognition receptors (PRRs).

[0005] PAMPs or MAMPs are invariant structures that are widely expressed among microbial taxa and have essential roles in microbial physiology. Only a very select group of molecules have been found to function as PAMPs. Conserved molecules from nematodes have also been found to trigger plant defenses and pathogen resistance. Thus, these molecules have been defined as nematode-associated molecular patterns (NAMPs). In addition to molecular patterns derived from pathogens, plants can also sense molecular patterns associated with cell wall disruption or cell damage, i.e., so-called danger / damage-associated molecular patterns (DAMPs).

[0006] PRRs are generally plasma membrane receptors that are usually coupled to an intracellular kinase domain or require a co-receptor to provide signal transduction function (Dangl et al., 2013, “Pivoting the plant immune system from dissection to deployment”, Science, 341(6147):746-751). Depending on the presence of a signal transduction domain, plant PRRs are classified as receptor-like kinases (RLKs) or receptor-like proteins (RLPs). The recognition of PAMPs, MAMPs, NAMPs, or DAMPs in the apoplast by pattern recognition receptors (PRRs) initiates a complex signal cascade that leads to PRR-triggered immunity (PTI). Adapted pathogens may be able to suppress the first layer of defense by secreting effector proteins that interfere with signal transduction (Jones and Dangl, 2006).

[0007] The second plant defense layer, effector-triggered immunity (ETI), relies on the specific recognition of effectors by disease resistance genes (Jones and Dangl, 2006). This recognition leads to a strong defense response, which is usually associated with local programmed cell death, the hypersensitive response. Since effectors are usually species- or isolate-specific, this second immune layer is only effective against isolates carrying the recognized effector, which is then referred to as an avirulence gene.

[0008] Whether a potential pathogen can overcome the first defense layer, PTI, and effectively multiply depends on its inherent ability to suppress the plant PTI response. But it also depends on the plant's ability to effectively and rapidly induce and, if necessary, maintain a defense response above a certain effective resistance threshold (Jones and Dangl, 2006).

[0009] The PTI response is generally conserved and includes the activation of mitogen-activated protein kinases (MAPKs), the production of reactive oxygen species, the activation of salicylic acid (SA)- and jasmonic acid (JA)-signaling pathways, and the enhanced expression of plant defense genes such as pathogenesis-related proteins. Transcriptional activation can usually be measured within minutes or hours after infection and decreases after an effective defense response.

[0010] Transcription of protein-coding genes in eukaryotes is orchestrated by RNA polymerase II (RNAPII), general transcription factors, mediator, and gene-specific transcription factors. The multi-subunit RNAPII is evolutionarily conserved from yeast to humans. Its largest subunit, Rpb1, contains a carboxy-terminal domain (CTD) composed of conserved heptapeptides with the consensus sequence Y1S2P3T4S5P6S7 repeats (Buratowski, 2009, “Progression through the RNA polymerase II CTD cycle”, MollCell, 36(4):541-546). The combinatorial complexity of CTD post-translational modifications constitutes a “CTD code” “read” by CTD-binding proteins to regulate the transcription cycle, modify chromatin structure, and regulate RNA capping, splicing, and polyadenylation. In particular, the CTD undergoes fluctuations in serine phosphorylation and dephosphorylation events regulated by various CTD kinases - usually cyclin-dependent kinases (CDKs) and phosphatases - during transcription initiation, elongation, and termination. Interactions between different CTD kinases and phosphatases provide a means to couple and coordinate specific stages of transcription by recruiting other factors desired for appropriate gene expression.

[0011] In Arabidopsis, the CTD phosphatase-like protein family has five members (CPL1-5) (Fukudome et al., 2014, “Arabidopsis CPL4 is an essential C-terminal domain phosphatase that suppresses xenobiotic stress responses”, Plant J, 80(1):27-39). It has been shown to have a preference for different phosphorylated serines in the heptapeptide repeat and has been shown to be involved in different biological processes.

[0012] Arabidopsis AtCPL1 is a negative regulator of stress-responsive gene expression under various abiotic stresses (cold, abscisic acid (ABA), salt treatment, and iron deficiency) (Koiwa et al., 2002, “C-terminal domain phosphatase-like family members (AtCPLs) differentially regulate Arabidopsis thaliana abiotic stress signalling, growth, and development”, PNAS, 99(16):10893-10898; Aksoy et al., 2013, “Loss of function of Arabidopsis C-terminal domain phosphatase-like 1 activates iron deficiency responses at the transcriptional level”, Plant Physiol, 161(1):330-345), and negatively regulates wound-induced JA-biosynthesis genes. Thatcher et al. (2018, “The Arabidopsis RNA Polymerase II Carboxyl Terminal Domain (CTD) Phosphatase-Like 1 (CPL1) is a biotic stress susceptibility gene”, Sci Rep, 8:13454) subsequently revealed that complete knockout of AtCPL1 in Arabidopsis led to enhanced resistance to necrotrophic fungal pathogens Fusarium oxysporum and Alternaria brassicicola, and reduced symptom development upon aphid (Myzus persicae) infestation. Additionally, Arabidopsis AtCPL1 mutants with premature stop codons (complete AtCPL1 knockout) showed delayed flowering.

[0013] Thatcher et al. (Sci Rep; 2018) did not describe plants with downregulated AtCPL1. To date, the effect of CPL genes on pathogen resistance in plant species other than Arabidopsis has not been described.

[0014] ERF922 is a plant ethylene response factor (ERF), which is a subfamily of the APETELA2 / ethylene response factor (AP2 / ERF) transcription factor superfamily in plants. The ERF922 mutant of rice was generated by CRISPR / Cas9 engineering, thereby enhancing rice blast resistance (Wang et al., 2016, “Enhanced Rice Blast Resistance by CRISPR / Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922”, PLoSONE, 11(4):e0154027). This publication showed that RNAi-mediated downregulation of ERF922 enhanced resistance to the fungal pathogen Magnaporthe grisea and acted as a negative regulator of plant defense. OsERF922 is a transcriptional activator that binds to the GCC element of the promoter and is induced by infection with Magnaporthe grisea (M. grisea). Downregulation of OsERF922 reduces the expression of defense-related genes including PR1 and P10. Summary of the Invention

[0016] The present invention relates to the induction or increase of pathogen resistance in plants, particularly achieved by reducing or eliminating the expression level of a negative regulator of plant defense genes or disease resistance genes. In certain embodiments, the present invention relates to a method for inducing or increasing pathogen resistance in plants by reducing or eliminating the expression level of CPL1 and / or ERF922 or affecting the activity or stability of these proteins. In fact, it has been found that both CPL1 and / or ERF922 are negative regulators of plant defense and are particularly suitable for the method of the present invention.

[0017] The inventors have hitherto identified previously unknown CPL1 and ERF922 genes in a variety of economically highly relevant crops, including maize, wheat, barley, rye, sorghum, potato, soybean, and sugar beet.

[0018] In addition, in several of these crops, multiple CPL1 and / or ERF922 genes have been identified, particularly some homologs and paralogs. In addition, differential expression of some homologs / paralogs has been found. The presence of such homologs or paralogs - especially when differentially expressed - clearly increases the level of complexity in many ways. However, advantageously, the presence of different paralogs allows for better fine-tuning of pathogen resistance.

[0019] The inventors have found that reducing the expression (or otherwise reducing the functionality) of CPL1 and / or ERF922 - especially CPL1 - rather than completely eliminating the expression can advantageously lead to increased pathogen resistance or tolerance without accompanying growth or developmental defects, such as growth delay in the case of ERF922 knockout or delayed flowering in the case of CPL1 knockout.

[0020] The above effects can be advantageously achieved using dominant negative forms of CPL1 and / or ERF922 - especially CPL1 - particularly in the presence of multiple homologs and / or paralogs, since a single dominant negative form can simultaneously inhibit all homologs and paralogs.

[0021] Furthermore, the inventors have for the first time demonstrated that reducing or eliminating the expression (or otherwise reducing or eliminating the functionality) of CPL1 and / or ERF922 can increase pathogen resistance or tolerance against biotrophic and hemibiotrophic pathogens (especially fungi), which operates in a completely different manner from necrotrophic pathogens.

[0022] Downregulation of negative regulators of plant defense such as CPL1 and ERF922 increases plant tolerance to diseases caused by fungal pathogens, bacteria, insects, and nematodes. For example, downregulation can be carried out by disrupting the coding sequence by CRISPR / nuclease or Tilling, reducing gene transcription by RNAi, miRNA, and by generating less functional alleles (e.g., by Tilling or CRISPR / nuclease).

[0023] The present invention is captured in particular by any one or more of statements 1 to 73 numbered below and any other statements and / or embodiments and any combination thereof.

[0024] 1. A method for increasing resistance and / or tolerance to pathogens in a plant, plant part, or plant population, comprising reducing or eliminating the expression, stability, and / or activity of CPL1 and / or ERF922 protein and / or gene in the plant, plant part, or plant population.

[0025] 2. The method according to statement 1, wherein the CPL1 and / or ERF922 gene is mutated.

[0026] 3. The method according to statement 1 or 2, wherein the coding sequence and / or regulatory sequence of the CPL1 and / or ERF922 gene is mutated.

[0027] 4. The method according to any one of statements 1 to 3, wherein the expression, stability, and / or activity of the CPL1 and / or ERF922 protein is reduced.

[0028] 5. The method according to any one of statements 1 to 4, comprising expressing a mutant CPL1 and / or ERF922 protein.

[0029] 6. The method according to statement 5, wherein the mutant CPL1 and / or ERF922 gene comprises a point mutation, preferably resulting in an amino acid substitution in the CPL1 and / or ERF922 protein.

[0030] 7. The method according to statement 5 or 6, wherein the mutant CPL1 and / or ERF922 protein is a dominant negative CPL1 and / or ERF922 protein.

[0031] 8. The method according to any one of statements 5 to 7, wherein the mutant CPL1 protein comprises a mutation in the DXDXT motif.

[0032] 9. The method according to any one of statements 5 to 8, wherein the mutant CPL1 protein comprises a mutation corresponding to D128X in AtCPL4, wherein X is an amino acid different from D.

[0033] 10. The method according to any one of statements 5 to 9, wherein the mutant CPL1 protein comprises a mutation corresponding to D128A in AtCPL4.

[0034] 11. The method according to any one of statements 5 to 10, wherein the mutant CPL1 protein comprises a mutation in the CPL1 protein corresponding to:

[0035] - D148X, preferably D148A, when the plant is from the genus Zea, preferably Zea mays;

[0036] - D149X, preferably D148A, when the plant is from the genus Zea, preferably Zea mays;

[0037] - D162X, preferably D162A, when the plant is from the genus Beta, preferably Beta vulgaris;

[0038] - D143X, preferably D143A, when the plant is from the genus Solanum, preferably Solanum tuberosum;

[0039] - D141X, preferably D141A, when the plant is from the genus Glycine, preferably Glycine max;

[0040] -D149X, preferably D149A, when the plant is from the genus Triticum, preferably Triticum aestivum;

[0041] -D147X, preferably D147A, when the plant is from the genus Triticum, preferably Triticum aestivum;

[0042] -D149X, preferably D149A, when the plant is from the genus Sorghum, preferably Sorghum bicolor;

[0043] wherein X is an amino acid different from D.

[0044] 12. The method according to any one of statements 1 to 11, wherein the wild-type CPL1 or ERF922 protein comprises a sequence that is preferably at least 95% identical to any one of the sequences of SEQ ID NO: 2-17 or 37-50 over its full length, or is encoded by a sequence that is preferably at least 95% identical to any one of the sequences of SEQ ID NO: 19-34 or 52-65 over its full length.

[0045] 13. The method according to any one of statements 1 to 12, wherein the pathogen is selected from fungi, bacteria, viruses, nematodes, and insects.

[0046] 14. The method according to any one of statements 1 to 13, wherein the pathogen is a biotrophic or hemibiotrophic pathogen.

[0047] 15. The method according to any one of statements 1 to 14, wherein the plant comprises at least two CPL1 and / or ERF922 genes.

[0048] 16. The method according to any one of statements 1 to 15, wherein the plant comprises at least two homologs or paralogs of CPL1 and / or ERF922.

[0049] 17. The method according to any one of statements 1 to 15, comprising reducing or eliminating the expression, stability, and / or activity of more than one CPL1 and / or more than one ERF922 protein in the plant.

[0050] 18. The method according to any one of statements 15 to 17, wherein the at least two CPL1 proteins are differentially expressed and / or wherein the at least two ERF922 proteins are differentially expressed.

[0051] 19. The method according to any one of statements 1 to 18, wherein the plant is a crop.

[0052] 20. The method according to any one of statements 1 to 19, wherein the plant is selected from the Gramineae family.

[0053] 21. The method according to any one of statements 1 to 20, wherein the plant is selected from the Pooideae subfamily.

[0054] 22. The method according to any one of statements 1 to 20, wherein the plant is selected from the genus Zea, Sorghum, Triticum, Hordeum, Secale, Beta, Glycine or Solanum.

[0055] 23. The method according to any one of statements 1 to 20, wherein the plant is selected from maize, sorghum, wheat, barley, rye, sugar beet, soybean or potato species.

[0056] 24. The method according to any one of statements 1 to 23, wherein the CPL1 and / or ERF922 protein and / or gene expression, stability and / or activity are reduced or eliminated by knocking out the CPL1 and / or ERF922 gene or knocking down the CPL1 and / or ERF922 protein.

[0057] 25. The method according to any one of statements 1 to 24, wherein the CPL1 and / or ERF922 protein and / or gene expression, stability and / or activity are reduced or eliminated by mutagenesis, RNAi or gene editing.

[0058] 26. The method according to any one of statements 1 to 24, wherein the method comprises (recombinantly or transgenically) introducing or introgressing a mutation in the CPL1 and / or ERF922 gene or a nucleotide sequence encoding a CPL1 and / or ERF922 having a mutation into the genome of a plant or plant part, preferably a mutation that results in a decrease or elimination of the mRNA expression of the gene and / or the expression of the CPL1 and / or ERF922 protein, a mutation that results in a CPL1 and / or ERF922 protein having a reduced activity upon translation, or a mutation that results in a CPL1 and / or ERF922 having a reduced stability.

[0059] 27. The method according to any one of statements 1 to 26, wherein the method comprises:

[0060] (a) (Recombinantly or transgenically) introducing or introgressing a mutation in the nucleotide sequence of the (endogenous (wild-type)) gene encoding CPL1 and / or ERF922 in a plant or plant part, preferably a mutation that results in a decrease or elimination of the (endogenous (full-length)) mRNA of the gene and / or the (endogenous (full-length)) CPL1 and / or ERF922 protein, a mutation that results in a CPL1 and / or ERF922 protein having a reduced activity upon translation, or a mutation that results in a CPL1 and / or ERF922 protein having a reduced stability;

[0061] (b) introducing or infiltrating into a plant or a plant part, recombinantly or transgenically, an RNAi molecule that targets, is complementary to, or hybridizes with a nucleotide sequence encoding the CPL1 and / or ERF922 protein, or a polynucleotide sequence encoding an RNAi molecule that targets, is complementary to, or hybridizes with a nucleotide sequence encoding the CPL1 and / or ERF922 protein; or

[0062] (c) introducing or infiltrating into a plant or a plant part, recombinantly or transgenically, an RNA-specific or DNA-specific CRISPR / Cas system that targets a nucleotide sequence encoding the CPL1 and / or ERF922 protein, and / or one or more polynucleotide sequences encoding said RNA-specific CRISPR / Cas system; or

[0063] (d) introducing or infiltrating into a plant or a plant part, recombinantly or transgenically, a chemical compound or an antibody that modifies the activity of the CPL1 and / or ERF922 protein when interacting with the CPL1 and / or ERF922;

[0064] (e) introducing or infiltrating into a plant or a plant part, recombinantly or transgenically, a dominant-negative CPL1 and / or ERF922 protein or one or more nucleic acids encoding a dominant-negative CPL1 and / or ERF922 protein;

[0065] (f) optionally, regenerating a plant from a plant part of any one of (a) to (d).

[0066] 28. The method according to any one of statements 1 to 27, wherein the plant is transgenic.

[0067] 29. A plant, a plant part, or a plant population, or progeny thereof, obtainable by the method according to any one of statements 1 to 28.

[0068] 30. A plant, a plant part, or a plant population having a reduced or eliminated expression, stability, and / or activity of the CPL1 and / or ERF922 protein and / or gene, as compared to the expression, stability, and / or activity in a plant, a plant part, or a plant population of the same species that does not have a reduced or eliminated expression, stability, and / or activity of the CPL1 and / or ERF922 protein and / or gene.

[0069] 31. The plant, the plant part, or the plant population according to statement 29 or 30, which is mutagenized.

[0070] 32. The plant, the plant part, or the plant population according to any one of statements 29 to 31, which is transgenic or gene-edited.

[0071] 33. A plant part according to any one of statements 1 to 32, which is a cell, tissue, organ, fruit or seed.

[0072] 34. A plant, plant part or plant population according to any one of statements 29 to 33, wherein the plant is a crop.

[0073] 35. A plant, plant part or plant population according to any one of statements 29 to 34, wherein the plant is selected from the Poaceae family.

[0074] 36. A plant, plant part or plant population according to any one of statements 29 to 35, wherein the plant is selected from the Pooideae subfamily.

[0075] 37. A plant, plant part or plant population according to any one of statements 29 to 34, wherein the plant is selected from the genus Zea, Sorghum, Triticum, Hordeum, Secale, Beta, Glycine or Solanum.

[0076] 38. A plant, plant part or plant population according to any one of statements 29 to 34, wherein the plant is selected from maize, sorghum, wheat, barley, rye, beet, soybean or potato.

[0077] 39. An (isolated) polynucleotide comprising a sequence that is preferably at least 90% identical to any one of the sequences of SEQ ID NO: 19 - 34 or 52 - 65 over its full length; or a sequence encoding a polypeptide that is preferably at least 90% identical to any one of the sequences of SEQ ID NO: 2 - 17, 80 - 87 or 37 - 50 over its full length.

[0078] 40. An (isolated) polynucleotide that specifically hybridizes to the polynucleotide of statement 39, its complement or its reverse complement.

[0079] 41. The (isolated) polynucleotide according to statement 40, wherein the polynucleotide is a primer or a probe.

[0080] 42. The (isolated) polynucleotide according to statement 40, wherein the polynucleotide is an RNAi polynucleotide, siRNA or shRNA.

[0081] 43. The (isolated) polynucleotide according to statement 40, wherein the polynucleotide is a guide RNA.

[0082] 44. A method for generating a plant or a plant part, comprising (a) providing a first plant according to any one of statements 29 to 38, (b) crossing the first plant with a second plant, (c) selecting a progeny plant having a reduced or eliminated expression, stability and / or activity of the CPL1 and / or ERF922 protein and / or gene as compared to the expression, stability and / or activity in plants of the same species, and optionally (d) harvesting the plant part from the progeny plant.

[0083] 45. Use of a (isolated) polynucleotide according to any one of statements 39 to 43 for increasing the resistance and / or tolerance of a plant, a plant part or a plant population to a pathogen and / or for generating a plant or a plant part or a plant according to any one of statements 29 to 38.

[0084] 46. The use according to statement 45, wherein the (isolated) polynucleotide encodes a polypeptide which is preferably at least 90% identical to any one of the sequences of SEQ ID NO: 80 - 87 over its full length.

[0085] 47. A plant, a plant part or a plant population comprising a polynucleotide according to any one of statements 39 to 43.

[0086] 48. The plant, the plant part or the plant population according to statement 47, wherein the plant, the plant part or the plant population is transgenic.

[0087] 49. The plant, the plant part or the plant population according to statement 47, wherein the plant, the plant part or the plant population recombinantly expresses the polynucleotide.

[0088] 50. The plant part according to any one of statements 47 to 49, which is a cell, a tissue, an organ, a fruit or a seed.

[0089] 51. A method for controlling pathogen infection in a plant (population), comprising:

[0090] a) providing a plant according to any one of statements 29 to 38 or 47 to 50 or growing a plant from a seed according to any one of statements 29 to 38 or 47 to 50,

[0091] b) cultivating the plant of a) under conditions of pathogen infection.

[0092] 52. The method according to statement 51, wherein pathogen infection is reduced.

[0093] 53. The method according to statement 51 or 52, wherein symptoms caused by the pathogen are reduced.

[0094] 54. The method according to any one of statements 51 to 53, wherein the conditions of pathogen infection include the presence of the pathogen.

[0095] 55. The method or application according to any one of statements 45 to 54, wherein the pathogen is selected from fungi, bacteria, viruses, nematodes and insects.

[0096] 56. The method or application according to any one of statements 45 to 55, wherein the pathogen is a biotrophic or hemibiotrophic pathogen.

[0097] 57. Use of the method according to any one of statements 1 to 28 in increasing plant yield (potential), preferably under conditions of pathogen infection.

[0098] 58. The use according to statement 52, wherein the yield is biomass or seed yield.

[0099] 59. The use according to statement 58, wherein the biomass is whole-plant biomass or biomass of plant parts.

[0100] 60. The use according to statement 59, wherein the plant parts are tissues, organs, fruits or seeds.

[0101] 61. The use according to statement 59 or 60, wherein the plant parts are harvestable plant parts.

[0102] 62. A method for producing feed or food having a reduced amount of fungal or bacterial toxins, comprising:

[0103] A) Controlling pathogen infection in a plant population by the method according to any one of statements 51 to 56,

[0104] B) Harvesting plant material from the population, and

[0105] C) Producing feed or food from the harvested plant material.

[0106] 63. Feed or food having a reduced content of fungal or bacterial toxins obtained by the method according to statement 62.

[0107] 64. The method or application according to any one of statements 45 to 63, wherein the plant is a crop.

[0108] 65. The method or application according to any one of statements 45 to 64, wherein the plant is selected from the Gramineae.

[0109] 66. The method or application according to any one of statements 45 to 65, wherein the plant is selected from the Pooideae.

[0110] 67. The method or application according to any one of statements 45 to 66, wherein the plant is selected from the genus Zea, Sorghum, Triticum, Hordeum, Secale, Beta, Glycine or Solanum.

[0111] 68. The method or application according to any one of statements 45 to 67, wherein the plant is selected from the species of corn, sorghum, wheat, barley, rye, sugar beet, soybean or potato.

[0112] 69. A method for identifying a plant, plant part or plant population having increased resistance and / or tolerance to a pathogen, comprising screening and / or identifying a mutation as defined in any one of statements 2 to 11.

[0113] 71. The method according to statement 69, further comprising the step of selecting a plant, plant part or plant population having a mutation as defined in any one of statements 2 to 11.

[0114] 72. A method for identifying a plant, plant part or plant population having increased resistance and / or tolerance to a pathogen, comprising screening and / or identifying a reduced or eliminated expression, activity and / or stability of the CPL1 and / or ERF922 protein and / or gene in the plant, plant part or plant population.

[0115] 73. The method according to statement 72, further comprising the step of selecting a plant, plant part or plant population having a reduced or eliminated expression, activity and / or stability of CPL1 and / or ERF922. Brief Description of the Drawings

[0117] Figure 1 : Sequence alignment between different protein sequences of the discovered CPL1 gene and comparison with the Arabidopsis thaliana AtCPL1 protein sequence.

[0118] Figure 2 : Sequence alignment between different coding sequences of the discovered CPL1 gene and comparison with the Arabidopsis thaliana AtCPL1 coding sequence.

[0119] Figure 3 : Vector diagram of the plasmid construct for maize transformation to silence the ZmCPL1 gene.

[0120] Figure 4 : Disease score of wheat leaves infected with Zymoseptoria tritici after VIGS-mediated downregulation of TaCPL1.

[0121] Figure 5 : Amino acid sequence comparison of ERF922-I and ERF922-II from different crops.

[0122] Figure 6 : Phylogenetic map of ERF922-I and ERF922-II genes from different crops.

[0123] Figure 7: Disease scores of wheat leaves infected with Zymoseptoria tritici after downregulation of TaERF922-I (TaERF922-3A-I, TaERF922-3B-I, TAERF922-3D-I) on chromosome 3 and TaERF922-II (TaERF922-2A-II, TaERF922-2B-II, TAERF922-2D-II) on chromosome 2 mediated by VIGS.

[0124] Figure 8 : Disease scores of wheat ears infected with Fusarium graminearum after downregulation of TaERF922-I (TaERF922-3A-I, TaERF922-3B-I, TAERF922-3D-I) on chromosome 3 and TaERF922-II (TaERF922-2A-II, TaERF922-2B-II, TAERF922-2D-II) on chromosome 2 mediated by VIGS.

[0125] Figure 9 : Vector diagram of a plasmid construct for maize transformation to silence the ZmERF922 gene. DETAILED DESCRIPTION OF THE INVENTION

[0127] Before describing the systems and methods of the present invention, it should be understood that the present invention is not limited to the specific systems and methods or combinations described, as such systems and methods and combinations can of course vary. It should also be understood that the terms used herein are not intended to be limiting, as the scope of the present invention will be limited only by the appended claims.

[0128] As used herein, the singular forms "a", "an" and "the" include both singular and plural referents unless the context clearly dictates otherwise.

[0129] As used herein, the terms "comprising" and "consisting of" are synonymous with "including" or "containing" and are inclusive or open-ended and do not exclude additional unrecited elements or method steps. It should be understood that the terms "comprising" and "consisting of" as used herein encompass the term "consisting of" as well as the term "consisting essentially of".

[0130] Numerical ranges limited by endpoints recited include all numbers and fractions within the corresponding ranges, as well as the recited endpoints.

[0131] As used herein, the term "about" or "approximate", when referring to a measurable value such as a parameter, quantity, duration, etc., means encompassing a specified value of + / - 20% or less, preferably + / - 10% or less, more preferably + / - 5% or less and even more preferably + / - 1% or less, provided that such variations are suitable for carrying out the invention as disclosed. It should be understood that the value itself to which the modifier "about" or "approximate" refers is also specifically and preferably disclosed.

[0132] Although the term "one or more" or "at least one", e.g., one or more members of a group of members or at least one member is clear in itself, by way of further illustration, the term includes in particular referring to any one of the said members, or any two or more of the said members, e.g., any ≥3, ≥4, ≥5, ≥6 or ≥7 etc. of the said members, and up to all of the said members.

[0133] All references cited in this specification are hereby incorporated by reference in their entirety. In particular, the teachings of all references specifically mentioned herein are incorporated by reference.

[0134] Unless otherwise defined, all terms used to disclose the present invention, including technical and scientific terms, shall have the meanings commonly understood by those of ordinary skill in the art to which the present invention pertains. Through further guidance, including term definitions, the teachings of the present invention are better understood.

[0135] Standard reference books that describe the general principles of recombinant DNA technology include Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1-3, ed. Sambrook et al., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; Current Protocols in Molecular Biology, ed. Ausubel et al., Greene Publishing and Wiley-Interscience, New York, 1992 (updated periodically) (“Ausubel et al. 1992”); the series Methods in Enzymology (Academic Press, Inc.); Innis et al., PCR Protocols: A Guide to Methods and Applications, Academic Press: San Diego, 1990; PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames and G.R. Taylor eds. (1995); Harlow and Lane, eds. (1988) Antibodies, a Laboratory Manual; and Animal Cell Culture (R.I. Freshney, ed. (1987). General principles of microbiology are described, for example, in Davis, B.D. et al., Microbiology, 3rd edition, Harper & Row, publishers, Philadelphia, Pa. (1980).

[0136] In the following paragraphs, different aspects of the invention are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects, unless the contrary is explicitly indicated. 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.

[0137] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment, but may be. Furthermore, in one or more embodiments, the particular features, structures, or characteristics may be combined in any suitable manner, as will be apparent to those skilled in the art from the present disclosure. Additionally, although some embodiments described herein include some features not included in other embodiments, combinations of features of different embodiments are meant to be within the scope of the invention and constitute different embodiments, as will be understood by those skilled in the art. For example, in the appended claims, any of the claimed embodiments can be used in any combination.

[0138] In the following detailed description of the invention, reference is made to the accompanying drawings which form a part hereof, and in which are shown by way of illustration specific embodiments in which the invention may be practiced. It is to be understood that other embodiments may be utilized and structural or logical changes may be made without departing from the scope of the invention. Accordingly, the following detailed description is not to be taken in a limiting sense, and the scope of the invention is defined by the appended claims.

[0139] Preferred statements (features) and embodiments of the invention are set forth below. Each statement and embodiment of the invention so defined can be combined with any other statement and / or embodiment, unless the contrary is explicitly indicated. In particular, any feature indicated as preferred or advantageous can be combined with any other one or more features or statements indicated as preferred or advantageous.

[0140] In one aspect, the invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part, or a plant population, comprising reducing or eliminating the expression, stability, and / or activity of a negative regulator of a plant defense or a plant pathogen resistance gene in the plant, the plant part, or the plant population.

[0141] In one embodiment, the negative regulator is CPL1 and / or ERF922. Thus, in one aspect, the invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part, or a plant population, comprising reducing or eliminating the expression, stability, and / or activity of CPL1 and / or ERF922 protein and / or gene in the plant, the plant part, or the plant population.

[0142] In a specific embodiment, the present invention includes a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression, stability and / or activity of CPL1 protein and / or gene in the plant, the plant part or the plant population.

[0143] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression, stability and / or activity of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0144] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression or activity of CPL1 and / or ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0145] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression or activity of CPL1 protein and / or gene in the plant, the plant part or the plant population.

[0146] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression or activity of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0147] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression of CPL1 and / or ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0148] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression of CPL1 protein and / or gene in the plant, the plant part or the plant population.

[0149] On the one hand, the present invention relates to a method for increasing resistance and / or tolerance to pathogens and / or increasing yield (potential) in a plant, a plant part or a plant population, comprising reducing or eliminating the expression of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0150] On the one hand, the present invention relates to a method for increasing the resistance and / or tolerance to pathogens and / or increasing the yield (potential) in plants, plant parts or plant populations, comprising reducing or eliminating the activity of CPL1 and / or ERF922 proteins and / or genes in plants, plant parts or plant populations.

[0151] On the one hand, the present invention relates to a method for increasing the resistance and / or tolerance to pathogens and / or increasing the yield (potential) in plants, plant parts or plant populations, comprising reducing or eliminating the activity of CPL1 protein and / or gene in plants, plant parts or plant populations.

[0152] On the one hand, the present invention relates to a method for increasing the resistance and / or tolerance to pathogens and / or increasing the yield (potential) in plants, plant parts or plant populations, comprising reducing or eliminating the activity of ERF922 protein and / or gene in plants, plant parts or plant populations.

[0153] On the one hand, the present invention relates to a method for increasing the resistance and / or tolerance to pathogens and / or increasing the yield (potential) in plants, plant parts or plant populations, comprising expressing dominant negative proteins and / or genes of CPL1 and / or ERF922 in plants, plant parts or plant populations.

[0154] On the one hand, the present invention relates to a method for increasing the resistance and / or tolerance to pathogens and / or increasing the yield (potential) in plants, plant parts or plant populations, comprising expressing dominant negative CPL1 protein and / or gene in plants, plant parts or plant populations.

[0155] On the one hand, the present invention relates to a method for increasing the resistance and / or tolerance to pathogens and / or increasing the yield (potential) in plants, plant parts or plant populations, comprising expressing dominant negative ERF922 protein and / or gene in plants, plant parts or plant populations.

[0156] On the one hand, the present invention relates to a method for generating or obtaining plants, plant parts or plant populations, comprising reducing or eliminating the expression, stability and / or activity of negative regulators of plant defense or plant pathogen resistance genes, more particularly CPL1 and / or ERF922 proteins and / or genes, in plants, plant parts or plant populations.

[0157] On the one hand, the present invention relates to a method for generating or obtaining plants, plant parts or plant populations, comprising reducing or eliminating the expression, stability and / or activity of CPL1 protein and / or gene in plants, plant parts or plant populations.

[0158] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression, stability and / or activity of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0159] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression or activity of CPL1 and / or ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0160] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression or activity of CPL1 protein and / or gene in the plant, the plant part or the plant population.

[0161] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression or activity of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0162] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression of CPL1 and / or ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0163] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression of CPL1 protein and / or gene in the plant, the plant part or the plant population.

[0164] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the expression of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0165] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the activity of CPL1 and / or ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0166] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the activity of CPL1 protein and / or gene in the plant, the plant part or the plant population.

[0167] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising reducing or eliminating the activity of ERF922 protein and / or gene in the plant, the plant part or the plant population.

[0168] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising expressing a CPL1 and / or an ERF922 dominant negative protein and / or gene in the plant, the plant part or the plant population.

[0169] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising expressing a CPL1 dominant negative protein and / or gene in the plant, the plant part or the plant population.

[0170] On the one hand, the present invention relates to a method for generating or obtaining a plant, a plant part or a plant population, comprising expressing an ERF922 dominant negative protein and / or gene in the plant, the plant part or the plant population.

[0171] The inventors have observed that in certain embodiments, the plant will contain more than one CPL1 and / or ERF922 gene. Thus, in certain embodiments, the expression, activity and / or stability of at least two CPL1 and / or at least two ERF922 proteins are reduced or eliminated.

[0172] In certain embodiments, the expression, activity and / or stability of at least two CPL1 and at least two ERF922 proteins are reduced or eliminated.

[0173] In certain embodiments, the expression, activity and / or stability of at least two CPL1 proteins are reduced or eliminated.

[0174] In certain embodiments, the expression, activity and / or stability of at least two ERF922 proteins are reduced or eliminated.

[0175] One or more mutations in one or more genes encoding negative regulators of plant defense or one or more plant pathogen resistance genes can lead to a decrease or elimination of the expression of the negative regulator in the plant, increasing or inducing pathogen resistance and / or yield. In certain embodiments, a nucleotide sequence of a gene having a mutation in a CPL1 and / or ERF922 gene or encoding a CPL1 and / or ERF922 having a mutation is introduced or introgressed (recombinantly or transgenically) into the genome of a plant or a plant part, preferably a mutation that results in a decrease or elimination of the expression of the gene's mRNA and / or CPL1 and / or ERF922 protein, a mutation that results in a CPL1 and / or ERF922 protein having reduced or eliminated activity upon translation (including dominant negative mutations), or a mutation that results in a CPL1 and / or ERF922 protein having reduced stability. Different methods are envisioned to ensure one or more mutations in one or more genes encoding negative regulators of plant defense or plant pathogen resistance genes. For example, mutations can be introduced by recombinant techniques or can be introduced by introgression of a gene from another plant (carrying one or more envisioned mutations).

[0176] In certain embodiments, the methods of the invention as described herein include:

[0177] (a) (Recombinantly or transgenically) introducing or introgressing a mutation in a plant or plant part in the nucleotide sequence of an (endogenous (wild-type)) gene encoding CPL1 and / or ERF922, preferably a mutation that results in reduced or eliminated expression of the (endogenous (full-length)) mRNA and / or (endogenous (full-length)) CPL1 and / or ERF922 protein of said gene, a mutation that results in a CPL1 and / or ERF922 protein with reduced activity upon translation, or a mutation that results in a CPL1 and / or ERF922 protein with reduced stability;

[0178] (b) (Recombinantly or transgenically) introducing or introgressing into the plant or plant part one or more RNAi molecules that target, are directed against, or hybridize to the nucleotide sequence encoding CPL1 and / or the nucleotide sequence encoding the ERF922 protein, or a polynucleotide sequence encoding one or more RNAi molecules that target, are directed against, or hybridize to the nucleotide sequence encoding CPL1 and / or the nucleotide sequence encoding the ERF922 protein;

[0179] (c) (Recombinantly or transgenically) introducing or introgressing into the plant or plant part one or more RNA-specific or DNA-specific CRISPR / Cas systems that target or are directed against the nucleotide sequence encoding CPL1 and / or the nucleotide sequence encoding the ERF922 protein, or one or more polynucleotide sequences encoding said RNA-specific or DNA-specific CRISPR / Cas systems;

[0180] (d) (Recombinantly or transgenically) introducing or introgressing into the plant or plant part a chemical compound or antibody (or polynucleic acid encoding the same) that modifies the activity of the CPL1 and / or ERF922 protein when interacting with said CPL1 and / or ERF922; or

[0181] (e) (Recombinantly or transgenically) introducing or introgressing into the plant or plant part a dominant negative CPL1 and / or ERF922 protein or one or more nucleic acids encoding a dominant negative CPL1 and / or ERF922 protein;

[0182] (f) Optionally, regenerating a plant from the plant part of any one of (a) to (d).

[0183] The methods described herein result in a plant, plant part, or plant population comprising one or more mutations in one or more genes encoding a negative regulator of a plant defense or plant pathogen resistance gene, resulting in reduced or eliminated expression of the negative regulator in the plant, plant part, or plant population, resulting in increased or induced pathogen resistance and / or yield in the plant, plant part, or plant population. Accordingly, the present invention provides plants, plant parts, and plant populations having increased or induced pathogen resistance and / or yield.

[0184] Accordingly, in one aspect, the present invention relates to a plant, plant part, or plant population obtained by any of the methods described above. In one aspect, the present invention relates to a plant obtained by any of the methods described above. In one aspect, the present invention relates to a plant part obtained by any of the methods described above. In one aspect, the present invention relates to a plant population obtained by any of the methods described above.

[0185] In one aspect, the present invention relates to a plant, plant part, or plant population that, for example, has reduced or eliminated expression, stability, and / or activity of CPL1 and / or ERF922 protein and / or gene compared to a wild-type plant, plant part, or plant population of the same species (or strain or genotype), or compared to a plant, plant part, or plant population of the same species (or strain or genotype) that does not have reduced or eliminated expression, stability, and / or activity of CPL1 and / or ERF922 protein and / or gene.

[0186] In one aspect, the present invention relates to a plant, plant part, or plant population comprising one or more polynucleotide sequences encoding CPL1 and / or ERF922 having a mutation, preferably a mutation that results in reduced or absent expression of the mRNA and / or protein, or a mutation that results in a truncated or non-functional protein upon translation.

[0187] In one aspect, the present invention relates to a plant, plant part, or plant population comprising one or more nucleotide sequences encoding CPL1 and / or ERF922 having reduced or eliminated mRNA and / or protein expression.

[0188] The present invention further provides plants, plant parts, and plant populations comprising a modified sequence of one or more genes encoding a negative regulator of a plant defense gene or a plant pathogen resistance gene. In one aspect, the present invention relates to a plant comprising:

[0189] (a) Mutations in the nucleotide sequence of the (endogenous (wild-type)) gene encoding CPL1 and / or ERF922, preferably mutations that result in reduced or eliminated expression of the (endogenous (full-length)) mRNA of the gene and / or the (endogenous (full-length)) CPL1 and / or ERF922 protein, mutations that result in CPL1 and / or ERF922 proteins with reduced activity upon translation, or mutations that result in CPL1 and / or ERF922 proteins with reduced stability;

[0190] (b) One or more RNAi molecules that target, bind to, or hybridize to the nucleotide sequence encoding CPL1 and / or the nucleotide sequence encoding the ERF922 protein, or a polynucleotide sequence encoding one or more RNAi molecules that target, bind to, or hybridize to the nucleotide sequence encoding CPL1 and / or the nucleotide sequence encoding the ERF922 protein;

[0191] (c) One or more RNA-specific or DNA-specific CRISPR / Cas systems that target or bind to the nucleotide sequence encoding CPL1 and / or the nucleotide sequence encoding the ERF922 protein, or one or more polynucleotide sequences encoding the RNA-specific or DNA-specific CRISPR / Cas systems;

[0192] (d) Chemical compounds or antibodies (or polynucleic acids encoding them) that alter the activity of the CPL1 and / or ERF922 protein when interacting with CPL1 and / or ERF922; or

[0193] (e) Dominant negative CPL1 and / or ERF922 proteins or one or more nucleic acids encoding the dominant negative CPL1 and / or ERF922 proteins.

[0194] In certain embodiments, plants are provided that comprise one or more negative regulators of a plant pathogen resistance gene or pathogen defense gene that have been knocked out and / or knocked down. Alternatively, the mutations in the negative regulators are dominant negative mutations such that the gene product negatively impacts the function of the wild-type negative regulator. In one aspect, the invention relates to plants comprising a knockout mutation of CPL1 and / or ERF922. In one aspect, the invention relates to plants comprising a knockout mutation of CPL1 and ERF922. In one aspect, the invention relates to plants comprising a knockout mutation of CPL1. In one aspect, the invention relates to plants comprising a knockout mutation of ERF922. In certain embodiments, the mutations are homozygous. In certain embodiments, the mutations are heterozygous.

[0195] On the one hand, the present invention relates to plants comprising knockdown mutations of CPL1 and / or ERF922. On the one hand, the present invention relates to plants comprising knockdown mutations of CPL1 and ERF922. On the one hand, the present invention relates to plants comprising a knockdown mutation of CPL1. On the one hand, the present invention relates to plants comprising a knockdown mutation of ERF922. In certain embodiments, the mutation is homozygous. In certain embodiments, the mutation is heterozygous.

[0196] On the one hand, the present invention relates to plants expressing a dominant negative CPL1 and / or ERF922 protein. On the one hand, the present invention relates to plants expressing a dominant negative CPL1 and ERF922 protein. On the one hand, the present invention relates to plants expressing a dominant negative CPL1. On the one hand, the present invention relates to plants expressing a dominant negative ERF922 protein. In certain embodiments, the mutation is homozygous. In certain embodiments, the mutation is heterozygous.

[0197] The term "plant" includes the whole plant, including its progeny or offspring. The term "plant part" includes any part or derivative of a plant, including specific plant tissues or structures, plant cells, plant protoplasts, plant cells or tissue cultures from which a plant can be regenerated, plant callus, plant clumps and plant cells that are intact in a plant or plant part, such as seeds, kernels, rachises, fruits, flowers, cotyledons, leaves, stems, buds, roots, root tips, straws, etc. Plant parts can include processed plant parts or derivatives, including flowers, oils, extracts, etc. In certain embodiments, the plant part or derivative as referred to herein is a cell, tissue or seed (or grain). In certain embodiments, the plant part or derivative as referred to herein is a seed (or grain). In certain embodiments, the plant part is a harvestable plant part, such as a seed or grain, fruit, root, leaf, stem, flower, tuber, bulb, rachis, etc.

[0198] In certain embodiments, the plant is a crop plant, such as a cash crop or a subsistence crop, such as a food or non-food crop, including an agricultural, horticultural, floral or cash crop. The term crop plant has its ordinary meaning known in the art. By way of further guidance, but not limited to, a crop is a plant grown by humans for food and other resources and can be widely planted and harvested for profit or livelihood, typically in an agricultural environment or context.

[0199] In certain embodiments, the plant is from the Poaceae family. As used herein, the term Poaceae refers to the family of grasses or Gramineae. Preferably, the Poaceae is a cereal (or grain grass), which is cultivated particularly for the edible components of its grains.

[0200] In certain embodiments, the plant is from the subfamily Pooideae. As used herein, the term subfamily Pooideae refers to the subfamily Pooideae within the family Poaceae. Preferably, the subfamily Pooideae is a cereal (or grain grass), particularly cultivated for the edible components of its grains. In certain embodiments, the plant is from the genus Zea, preferably maize (Zea mays). In certain embodiments, the plant is from the genus Sorghum, preferably sorghum (Sorghum bicolor). In certain embodiments, the plant is from the genus Triticum, preferably wheat (Triticum aestivum). In certain embodiments, the plant is from the genus Hordeum, preferably barley (Hordeum vulgare). In certain embodiments, the plant is from the genus Secale, preferably rye (Secale cereale). In certain embodiments, the plant is from the genus Beta, preferably sugar beet (Beta vulgaris). In certain embodiments, the plant is from the genus Glycine, preferably soybean (Glycine max). In certain embodiments, the plant is from the genus Solanum, preferably potato (Solanum tuberosum). In certain embodiments, the plant is from the genus Oryza, preferably rice (Oryza sativa).

[0201] In certain embodiments, the plant part is or comprises propagation material. In certain embodiments, the plant part or derivative is not (functional) propagation material, such as germplasm, seeds or plant embryos or other materials from which a plant can be regenerated. In certain embodiments, the plant part or derivative does not comprise (functional) male and female reproductive organs. In certain embodiments, the plant part or derivative is or comprises propagation material, but is not used or cannot (any longer) be used as propagation material for the production or generation of new plants, such as propagation material that has been rendered non-functional chemically, mechanically or otherwise, for example by heat treatment, acid treatment, compaction, fragmentation, shredding, etc.

[0202] As used herein, the term "plant population" may be used interchangeably with a population of plants. A plant population preferably comprises a large number of individual plants, for example preferably at least 10 species, such as 20, 30, 40, 50, 60, 70, 80 or 90 species, more preferably at least 100 species, such as 200, 300, 400, 500, 600, 700, 800 or 900 and even more preferably at least 1000 species, such as at least 10000 or at least 100000 plant species.

[0203] As used herein, the terms "increased pathogen tolerance" and "increased pathogen resistance" relate to any alleviation of any symptom of pathogen infection (such as damage or loss of biomass), any reduction in symptom presentation, any improvement in symptoms or any combination thereof. The increased pathogen resistance or tolerance as referred to herein may also relate to the ability of a plant to maintain, for example, its biomass production (such as harvestable biomass production, such as seed yield) during or upon pathogen infection. A pathogen-resistant or -tolerant plant, plant cell or plant part may herein refer to a plant, plant cell or plant part that has increased resistance / tolerance to a pathogen as compared to its parent plant (and that does not have reduced or eliminated expression, stability and / or activity of the CPL1 and / or ERF922 protein and / or gene). The resistance herein may relate to the ability of a plant to restrict pathogen reproduction. Herein, tolerance may relate to the ability of a plant to reduce the impact of infection on its fitness, regardless of the level of pathogen reproduction. Methods for determining pathogen resistance / tolerance are known to those skilled in the art, such as visual scoring of pathogen infection or pathogen-induced damage, determination of biomass (yield), etc. As used herein, the terms "increased pathogen tolerance" and "increased pathogen resistance" may be used interchangeably with "reduced sensitivity" or "reduced susceptibility" to a pathogen. Thus, a plant, plant part or plant population that is more resistant or more tolerant to a pathogen according to the invention is considered to be less sensitive to such a pathogen. When used herein, less sensitive or less susceptible may be regarded as "more tolerant" or "more resistant". Similarly, "more tolerant" or "more resistant" may be regarded as "less sensitive" or "less susceptible", and vice versa. When used herein, more sensitive or more susceptible may be regarded as "less tolerant" or "less resistant", and vice versa. Likewise, "less tolerant" or "less resistant" may be regarded as "more sensitive" or "more susceptible", and vice versa.

[0204] A plant, plant part or plant population having increased pathogen resistance or tolerance as described herein may be used to control pathogen infestation or infection. Thus, in one aspect, the invention relates to the use of such a plant to control pathogen infestation or infection. Preferably, pathogen infestation or infection is controlled by reducing pathogen infestation or infection or reducing the symptoms of pathogen infestation or infection at the plant, plant part or plant population level, as further described below.

[0205] In certain embodiments, the increased resistance or tolerance may itself be manifested as a reduction in infection or infestation at the (sub)plant level (e.g., specific cells, organs or tissues, such as the harvestable parts of the plant or e.g. leaves, stems, fruits or seeds) or at the population level, e.g., a reduction in the amount of pathogen (e.g., in per plant area or per plant biomass), a reduction in the reproduction (rate) or spread (rate) / distribution of the pathogen, and a reduction in the rate of pathogen spread at a particular time during the (growing) season, e.g., a reduction of at least 5%, preferably at least 10%, e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or (about) 100%. At the population level, the increased resistance or tolerance may itself be manifested as a reduction in infection or infestation as described above, but may also be manifested as, e.g., a reduction in the number of infected plants (or combinations thereof), e.g., a reduction of at least 5%, preferably at least 10%, e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or (about) 100%. It should be understood that such a reduction in infection or infestation may be relative to a reference plant (part) or population (e.g., a corresponding wild-type plant not according to the invention).

[0206] In certain embodiments, the increased resistance or tolerance may itself be manifested as a reduction in biomass or yield loss, or as a reduction in specific (harvestable) plant parts (e.g., the amount or weight of seeds or fruits) due to or as a result of pathogen infection. In certain embodiments, resistant or tolerant plants show a lower loss of biomass production (e.g., expressed as g / day or kg / ha or kg / ha / day, e.g., expressed in dry matter, such as in weight percentage) of at least 1%, preferably at least 2%, e.g., at least 3%, at least 4%, at least 5%, e.g., at least 10%, at least 15%, or at least 20% or more under pathogen infection compared to corresponding control plants, such as plants with lower resistance or tolerance or plants not according to the invention as described herein. In certain embodiments, resistant or tolerant plants show a higher biomass production (e.g., expressed as g / day or kg / ha or kg / ha / day, e.g., expressed in dry matter, such as in weight percentage) of at least 1%, preferably at least 2%, e.g., at least 3%, at least 4%, at least 5%, e.g., at least 10%, at least 15%, or at least 20% or more under pathogen infection compared to corresponding control plants, such as plants with lower resistance or tolerance or plants not according to the invention as described herein.

[0207] As used herein, the term "yield potential" refers to the maximum yield obtainable at harvest.

[0208] As used herein, the term "pathogen" generally refers to any type of infectious agent capable of causing (infectious) disease and includes, but is not limited to, viruses, bacteria, protozoa, prions, viroids, or fungi (including yeasts). Parasites such as insects or worms, as well as parasitic plants or algae, are generally also included within the term pathogen as used herein.

[0209] In certain embodiments, the pathogen is a biotrophic pathogen. In certain embodiments, the pathogen is a hemibiotrophic pathogen. In certain embodiments, the pathogen is not a necrotrophic pathogen.

[0210] Biotrophic pathogens obtain energy from living cells; they are found on or within living plants, can have very complex nutritional requirements, and do not (rapidly) kill the host plant. In contrast, necrotrophic pathogens obtain energy from killed cells; they rapidly invade and kill plant tissue and then live saprophytically on the dead remains. Hemibiotrophic pathogens have an initial biotrophic phase followed by a necrotrophic phase.

[0211] As used herein, a reduced (protein and / or gene / mRNA) expression level can refer to a reduction in the expression level of about at least 10%, preferably at least 30%, more preferably at least 50%, such as at least 20%, 40%, 60%, 80% or more, such as at least 85%, at least 90%, at least 95% or more. If the expression level is reduced by at least 80%, preferably at least 90%, more preferably at least 95%, then the expression is (substantially) absent or eliminated. In certain embodiments, if a protein and / or mRNA cannot be detected, particularly a wild-type or native protein and / or mRNA, then the expression is (substantially) absent. In certain embodiments, the protein and / or gene / mRNA expression level is reduced by 20 - 80%, such as 30 - 70% or 40 - 60%, such as (about) 50%, compared to the expression level in a reference plant, which can be a wild-type plant or a plant that does not contain a mutated CPL1 and / or ERF822 or any (genetic) event that results in reduced expression, as described elsewhere herein. The expression level can be determined by any means known in the art, such as by standard detection methods including, for example, (quantitative) PCR, northern blotting, western blotting, ELISA, etc.

[0212] The reduced expression level may be due to increased turnover of the mRNA or protein, such as increased degradation or reduced stability. The reduced expression level may be due to a reduced expression rate or a reduced transcription rate. The reduced expression level may be due to a reduced copy number (e.g., heterozygous wild-type and mutant CPL1 and / or ERF922).

[0213] As used herein, a reduced expression rate can refer to the expression rate of a nucleotide sequence being reduced by more than 10%, 15%, 20%, 25% or 30% compared to a specified reference value, preferably more than 40%, 45%, 50%, 55%, 60% or 65%, more preferably more than 70%, 75%, 80%, 85%, 90%, 92%, 94%, 96% or 98%, the reference being, for example, a plant that does not contain the genetic modification or other modification according to the invention as described elsewhere herein, or a reference plant such as a wild-type plant. However, this may also mean that the expression rate of the nucleotide sequence is reduced by 100%. A reduction in the expression rate preferably results in a change in the phenotype of the plant with the reduced expression rate.

[0214] As used herein, a reduced transcription rate can refer to the transcription rate of a nucleotide sequence being reduced by more than 10%, 15%, 20%, 25% or 30% compared to a specified reference, preferably more than 40%, 45%, 50%, 55%, 60% or 65%, more preferably more than 70%, 75%, 80%, 85%, 90%, 92%, 94%, 96% or 98%, the reference being, for example, a plant that does not contain the genetic modification or other modification according to the invention as described elsewhere herein, or a reference plant such as a wild-type plant. However, this may also mean that the transcription rate of the nucleotide sequence is reduced by 100%. A reduction in the transcription rate preferably results in a change in the phenotype of the plant with the reduced transcription rate.

[0215] As used herein, a reduced (protein) activity can mean that the activity is reduced by at least about 10%, preferably at least 30%, more preferably at least 50%, such as at least 20%, 40%, 60%, 80% or more, such as at least 85%, at least 90%, at least 95% or more. If the activity is reduced by at least 80%, preferably at least 90%, more preferably at least 95%, then the activity is (substantially) absent or eliminated. In certain embodiments, the activity is (substantially) absent if no activity can be detected, especially wild-type or native protein activity. The (protein) activity level can be determined by any means known in the art, depending on the type of protein, for example, by standard assay methods, including, for example, enzyme assays (for enzymes), transcription assays (for transcription factors), assays for analyzing phenotypic outputs, etc.

[0216] As used herein, reduced stability can refer to reduced protein stability or reduced RNA stability such as mRNA stability. The stability of a protein or RNA can be determined by methods known in the art, such as determining the protein / RNA half-life. In certain embodiments, a reduction in protein or RNA stability means a reduction in stability of at least about 10%, preferably at least 30%, more preferably at least 50%, such as at least 20%, 40%, 60%, 80% or more, such as at least 85%, at least 90%, at least 95%. In certain embodiments, a reduced protein or RNA stability means a reduction in half-life of at least about 10%, preferably at least 30%, more preferably at least 50%, such as at least 20%, 40%, 60%, 80% or more, such as at least 85%, at least 90%, at least 95%, such as a 2-fold, 3-fold, 4-fold, 5-fold or greater reduction in half-life. Stability can be compared to a reference value as defined above.

[0217] Expression levels, stability or activity can be compared between different plants (or plant parts), such as a plant (part) having reduced CLP1 and / or ERF922 expression, stability or activity according to the present invention compared to a reference plant (part) such as a wild-type plant (part). Expression levels, stability or activity can be compared between different conditions. Expression levels, stability or activity can be compared to a predetermined threshold. Such a predetermined threshold can correspond, for example, to the expression level, stability or activity of a particular genotype or under particular conditions.

[0218] In certain embodiments, the expression, stability and / or activity of CPL1 and / or ERF922 protein and / or gene is reduced or eliminated by knocking out the CPL1 and / or ERF922 gene or knocking down the CPL1 and / or ERF922 protein, as described elsewhere herein. In certain embodiments, as described elsewhere herein, the expression, stability and / or activity of CPL1 and / or ERF922 protein and / or gene is reduced or eliminated by mutagenesis, RNAi or gene editing.

[0219] It should be understood that reduced expression, activity or stability preferably refers to reduced expression, activity or stability of wild-type (functional) CPL1 or ERF922. For example, a knockout generated by genomic insertion or creation of a premature stop codon can result in the expression of a truncated mRNA / protein whose expression level is similar to that of the non-mutated mRNA / protein. However, the resulting protein has an affected functionality and thus the expression (and in this case the activity) is considered reduced. Similarly, introduction of a dominant negative variant of a protein may not affect the expression of the endogenous wild-type (functional) protein. However, at least the wild-type (functional) protein activity is reduced due to the presence of the dominant negative variant.

[0220] As used herein, the term "dominant negative" has its ordinary meaning known in the art. By way of further guidance, but not limited to, a dominant negative protein contains a mutation in its gene product that has an adverse effect on the normal wild-type gene product within the same cell. This typically occurs if the product can still interact with the same elements as the wild-type product, but blocks some aspect of its function, such as, for example: a mutation in a transcription factor that removes the activation domain but still contains the DNA-binding domain (the product can then block the binding of the wild-type transcription factor to the DNA site, resulting in a reduced level of gene activation); a mutation in an enzyme that abolishes catalytic activity (the product can then block the wild-type by binding and thus tittering out the substrate without catalytic conversion); a protein with dimerization function (a mutation that removes the functional domain but retains the dimerization domain results in a dominant negative phenotype because some of the protein dimers will lack a functional domain); and so on.

[0221] A decrease in expression, activity, and / or stability as described herein can be constitutive or conditional, such as inducible and / or tissue-specific (e.g., in the reproductive organs). Means for conditional manipulation, such as inducible or tissue-specific, are well known in the art.

[0222] As used herein, the terms "polypeptide" or "protein" (the two terms are used interchangeably herein) refer to a peptide, protein, or polypeptide comprising an amino acid chain of a given length, wherein the amino acid residues are joined by covalent peptide bonds. However, peptidomimetics of such proteins / polypeptides in which the amino acids and / or peptide bonds have been replaced by functional analogs are also included in the present invention, as well as in addition to the 20 genetically encoded amino acids such as selenocysteine. Peptides, oligopeptides, and proteins can be referred to as polypeptides. The term polypeptide also refers to, but does not exclude, modifications of the polypeptide, such as glycosylation, acetylation, phosphorylation, etc. Such modifications are well described in basic textbooks and more detailed monographs as well as in the research literature.

[0223] Amino acid substitutions include amino acid changes in which an amino acid is replaced with a different naturally occurring amino acid residue. Such substitutions can be classified as "conservative<1>", in which the amino acid residue contained in the wild-type protein is replaced with another naturally occurring amino acid having similar characteristics, such as Gly<>Ala, Val<>Ile<>Leu, Asp<>Glu, Lys<>Arg, Asn<>Gln, or Phe<>Trp<>Tyr. Substitutions encompassed by the present invention can also be "non-conservative", in which the amino acid residue present in the wild-type protein is replaced with an amino acid having different properties, such as a naturally occurring amino acid from a different group (e.g., replacing a charged amino acid with alanine). As used herein, "similar amino acids" refers to amino acids having similar amino acid side chains, i.e., amino acids having polar, non-polar, or nearly neutral side chains. As used herein, "dissimilar amino acids" refers to amino acids having different amino acid side chains, e.g., an amino acid having a polar side chain is dissimilar to an amino acid having a non-polar side chain. Polar side chains generally tend to be present on the surface of a protein, where they can interact with the aqueous environment in a cell ("hydrophilic" amino acids). On the other hand, "non-polar" amino acids tend to be located in the center of a protein, where they can interact with similar non-polar neighbors ("hydrophobic" amino acids). Examples of amino acids having polar side chains are arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, histidine, lysine, serine, and threonine (all are hydrophilic amino acids except cysteine, which is a hydrophobic amino acid). Examples of amino acids having non-polar side chains are alanine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, and tryptophan (all are hydrophobic amino acids except glycine, which is a neutral amino acid).

[0224] As used herein, the term "gene" refers to a polymeric form of nucleotides of any length, which are ribonucleotides or deoxyribonucleotides. The term includes double-stranded and single-stranded DNA and RNA. Also included are known types of modifications, such as methylation, "capping", and replacement of one or more naturally occurring nucleotides with analogs. Preferably, a gene contains a coding sequence encoding a polypeptide as defined herein. A "coding sequence" is a nucleotide sequence that, when placed under or in the control of appropriate regulatory sequences, is transcribed into mRNA and / or translated into a polypeptide. The boundaries of the coding sequence are determined by a translation initiation codon at the 5'-end and a translation termination codon at the 3'-end. The coding sequence can include, but is not limited to, mRNA, cDNA, recombinant nucleic acid sequences, or genomic DNA, and in some cases, introns may also be present.

[0225] As used herein, the term "endogenous" refers to a gene or allele that is present at its native genomic location. The term "endogenous" can be used interchangeably with "native". However, this does not exclude the existence of one or more nucleic acid differences from the wild-type allele. In certain embodiments, the difference from the wild-type allele can be limited to less than 9, preferably less than 6, more particularly less than 3 nucleotide differences. More particularly, the difference from the wild-type sequence can be in only one nucleotide. In certain embodiments, the difference from the wild-type allele can be limited to less than 9, preferably less than 6, more particularly less than 3 nucleotide differences. More particularly, the difference from the wild-type sequence can be in only one nucleotide. Preferably, the endogenous allele encodes a modified protein having less than 9, preferably less than 6, more particularly less than 3, and even more preferably only one amino acid difference from the wild-type protein. In certain embodiments, the endogenous gene or allele is a wild-type gene or allele. In certain embodiments, the endogenous gene is mutagenized as described elsewhere herein. In certain embodiments, the expression, activity, and / or stability of the endogenous gene is modified as described elsewhere herein.

[0226] As used herein, the term "homozygous" refers to a single cell or plant that has the same alleles at one or more or all loci. When the term is used with respect to a particular locus or gene, it means that at least that locus or gene has the same alleles. As used herein, the term "homozygous" refers to the genetic condition that exists when the same alleles are located at the corresponding loci on homologous chromosomes. As used herein, the term "heterozygous" refers to a single cell or plant that has different alleles at one or more or all loci. When the term is used with respect to a particular locus or gene, it means that at least that locus or gene has different alleles. As used herein, the term "heterozygous" refers to the genetic condition that exists when different alleles are located at the corresponding loci on homologous chromosomes. In certain embodiments, the CPL1 and / or ERF922 genes as described herein are homozygous (e.g., all mutated CPL1 and / or ERF922 on homologous or homologous chromosomes). In certain embodiments, the CPL1 and / or ERF922 genes as described herein are heterozygous (e.g., (at least) one mutated CPL1 and / or ERF922 and (at least) one wild-type CPL1 and / or ERF922 on homologous or homologous chromosomes).

[0227] "Polymorphism" refers to DNA variation between two or more individuals in a population. The polymorphism preferably has a frequency of at least 1% in the population. Useful polymorphisms can include single nucleotide polymorphisms (SNPs), simple sequence repeats (SSRs), or insertion / deletion polymorphisms, also referred to herein as "insertions / deletions (indels)". The term "insertion / deletion" refers to an insertion or deletion where one sequence can be said to have an inserted nucleotide or DNA fragment relative to a second sequence, or the second sequence can be said to have a deleted nucleotide or DNA fragment relative to the first sequence.

[0228] The term "sequence" as used herein refers to nucleotide sequences, polynucleotides, nucleic acid sequences, nucleic acids, nucleic acid molecules, peptides, polypeptides, and proteins, depending on the context in which the term "sequence" is used. The terms "nucleotide sequence", "polynucleotide", "nucleic acid sequence", "nucleic acid", "nucleic acid molecule" are used interchangeably herein and refer to nucleotides in a polymeric, unbranched form of any length, which are ribonucleotides or deoxyribonucleotides or a combination of both. Nucleic acid sequences include DNA, cDNA, genomic DNA, RNA, synthetic forms, and mixed polymers, sense and antisense strands, or may contain non-natural or derivatized nucleobases, which will be readily understood by those skilled in the art.

[0229] As used herein, the term "sequence identity" refers to the degree of identity between any given nucleic acid sequence and a target nucleic acid sequence. The percent sequence identity is calculated by determining the number of matching positions in the aligned nucleic acid sequences, dividing the number of matching positions by the total number of aligned nucleotides, and then multiplying by 100. A matching position is a position where the same nucleotide appears at the same position in the aligned nucleic acid sequences. The percent sequence identity of any amino acid sequence can also be determined. To determine the percent sequence identity, the BLAST2Sequences (Bl2seq) program from BLASTZ, which is an independent version of BLASTN and BLASTP, is used to compare the target nucleic acid or amino acid sequence with the identified nucleic acid or amino acid sequence. This independent version of BLASTZ can be obtained from the Fish & Richardson website (www.fr.com / blast) or the National Center for Biotechnology Information website of the US government (www.ncbi.nlm.nih.gov). Instructions on how to use the Bl2seq program can be found in the README file accompanying BLASTZ. BI2seq uses the BLASTN or BLASTP algorithm to compare two sequences.

[0230] BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. To compare two nucleic acid sequences, the options are set as follows: -i is set to the file containing the first nucleic acid sequence to be compared (e.g., C:\seq l.txt); -j is set to the file containing the second nucleic acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastn; -o is set to any desired file name (e.g., C:\output.txt); -q is set to -1; -r is set to 2; all other options are kept at their default settings. The following command will generate an output file containing the comparison between the two sequences: C:\B12seq -i c:\seql.txt -j c:\seq2.txt -p blastn -o c:\output.txt -q -1 -r 2. If the target sequence has homology to any part of the identified sequence, the specified output file will present these homologous regions as aligned sequences. If the target sequence has no homology to any part of the identified sequence, the specified output file will not present aligned sequences. Once aligned, the length is determined by counting the number of consecutive nucleotides in the target sequence that align with the identified sequence starting from any matching position and ending at any other matching position. A matching position is any position where the same nucleotide is present in both the target sequence and the identified sequence. Gaps that occur in the target sequence are not counted because gaps are not nucleotides. Similarly, gaps that are present in the identified sequence are not counted because the target sequence nucleotides are being counted, not the nucleotides from the identified sequence. The percentage identity over a particular length is determined by counting the number of matching positions over that length, dividing that number by the length, and then multiplying the resulting value by 100. For example, if (i) a 500-base nucleic acid target sequence is compared to a subject nucleic acid sequence, (ii) the Bl2seq program presents 200 bases of the target sequence that align with a region of the subject sequence, where the first and last bases of the 200-base region are matching, and (iii) the number of matches over these 200 aligned bases is 180, then the 500-base nucleic acid target sequence has a length of 200 bases and 90% sequence identity over that length (i.e., 180 / 200 × 100 = 90). It should be understood that different regions within a single nucleic acid target sequence that align with the identified sequence can each have their own percentage identity. Note that the value of the percentage identity is rounded to the nearest tenth. For example, 78.11, 78.12, 78.13, and 78.14 are rounded down to 78.1, while 78.15, 78.16, 78.17, 78.18, and 78.19 are rounded up to 78.2. Also note that the length value will always be an integer.

[0231] "Isolated nucleic acid" shall be understood as nucleic acid isolated from its natural or original environment. The term also includes synthetically produced nucleic acid. "Isolated nucleic acid sequence" or "isolated DNA" refers to a nucleic acid sequence that no longer exists in the natural environment from which it was isolated, such as a nucleic acid sequence in a bacterial host cell or a plant nuclear or plastid genome. When "sequence" is referred to herein, it should be understood that a molecule having such a sequence refers to, for example, a nucleic acid molecule. "Host cell" or "recombinant host cell" or "transformed cell" is a term for a new individual cell (or organism) that has arisen because at least one nucleic acid molecule has been introduced into said cell. The host cell is preferably a plant cell or a bacterial cell. The host cell may contain the nucleic acid as an extrachromosomal (episomal) replicating molecule, or contain nucleic acid integrated into the nuclear or plastid genome of the host cell, or as an introduced chromosome such as a minichromosome.

[0232] In one embodiment, when a nucleic acid sequence (e.g., DNA or genomic DNA) is said to have "substantial sequence identity" with a reference sequence or to have at least 80% or more sequence identity with the reference sequence, such as at least 85%, 90%, 95%, 98% or more or 99% or more nucleic acid sequence identity, the nucleotide sequence is considered to be substantially the same as the given nucleotide sequence and can be identified using stringent hybridization conditions. In another embodiment, the nucleic acid sequence contains one or more mutations compared to the given nucleotide sequence, but can still be identified using stringent hybridization conditions. "Stringent hybridization conditions" can be used to identify nucleotide sequences that are substantially the same as a given nucleotide sequence. Stringent conditions depend on the sequence and will vary in different circumstances. Typically, stringent conditions are selected to be about 5°C below the thermal melting temperature (Tm) of a particular sequence at a defined ionic strength and pH. Tm is the temperature at which 50% of the target sequence hybridizes to a perfectly matched probe (at a defined ionic strength and pH). Typical stringent conditions will be selected such that the salt concentration is about 0.02 mol, pH 7 and the temperature is at least 60°C. Decreasing the salt concentration and / or increasing the temperature increases the stringency. Stringent conditions for RNA-DNA hybridization (using, for example, a 100 nt probe in a Northern blot) are, for example, those conditions that include washing at least once for 20 minutes in 0.2× SSC at 63°C or equivalent conditions. Stringent conditions for DNA-DNA hybridization (using, for example, a 100 nt probe in a Southern blot) are, for example, those conditions that include washing at least once (usually 2 times) for 20 minutes at a temperature of at least 50°C, usually about 55°C in 0.2× SSC or equivalent conditions. See also Sambrook et al. (1989) and Sambrook and Russell (2001).

[0233] "Stringent hybridization conditions" can be used to identify nucleotide sequences that are substantially identical to a given nucleotide sequence. Stringent conditions depend on the sequence and vary in different circumstances. Typically, stringent conditions are selected to be about 5°C lower than the thermal melting temperature (Tm) of the specific sequence at a defined ionic strength and pH. The Tm is the temperature at which 50% of the target sequence hybridizes to a perfectly matched probe (at a defined ionic strength and pH). Typical stringent conditions will be selected where the salt concentration is about 0.02 M, the pH is 7 and the temperature is at least 60°C. Decreasing the salt concentration and / or increasing the temperature increases the stringency. Stringent conditions for RNA-DNA hybridization (e.g., Northern blotting using a 100 nt probe) are, for example, those that include at least one wash for 20 minutes in 0.2×SSC at 63°C or equivalent conditions. Stringent conditions for DNA-DNA hybridization (e.g., Southern blotting using a 100 nt probe) are, for example, those that include at least one wash (usually two) for 20 minutes at a temperature of at least 50°C, typically about 55°C, in 0.2×SSC or equivalent conditions. See also Sambrook et al. (1989) and Sambrook and Russell (2001).

[0234] As used herein, a "functional fragment" of a nucleotide sequence refers to a nucleotide sequence fragment having the same or comparable functionality as the complete nucleotide sequence from which the functional fragment is derived. Thus, a functional fragment can have a nucleotide sequence that is the same or homologous to the complete nucleotide sequence over at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 94%, 96%, 97%, 98% or 99% of its length. Additionally, a "functional fragment" of a nucleotide sequence can also refer to a nucleotide sequence fragment that alters the functionality of the total nucleotide sequence, such as during the process of post-transcriptional gene silencing. Thus, a functional fragment of a nucleotide sequence can comprise at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25, preferably at least 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 120 or 140, more preferably at least 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900 or 1000 consecutive nucleotides of the complete nucleotide sequence.

[0235] The "functional portion" of a protein refers to a protein fragment or a fragment of the amino acid sequence encoding the protein, wherein the fragment can perform the same or comparable function as the whole protein in a plant cell. The functional portion of the protein has the same amino acid sequence as the protein from which the functional portion is derived over at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 94%, 96%, 97%, 98% or 99% of its length or - with conservative and semi-conservative amino acid substitutions - a similar amino acid sequence.

[0236] As used herein, "CPL1" refers to RNA polymerase II C-terminal domain phosphatase-like 1. By way of example and not limitation, CPL1 as used herein can refer to any ortholog, homolog, paralog or homolog of Arabidopsis thaliana CPL1, as represented by the protein sequence of SEQ ID NO:1 or the coding sequence / cDNA sequence of SEQ ID NO:18.

[0237] In certain embodiments, a plant (or plant part or population) contains more than one CPL1 gene, which can be homologous genes or paralogous genes, or both. In certain embodiments, a plant (or plant part or population) contains two or more CPL1 genes, which can be homologous genes or paralogous genes, or both. In certain embodiments, different CPL1 homologs or paralogs are differentially expressed. Differential expression may require expression in different cells, tissues or organs; expression at different growth stages; or both.

[0238] As used herein, "ERF922" refers to ethylene response factor 922, also known as ethylene response factor 922. By way of example but not limitation, ERF922 as used herein can refer to any ortholog, homolog, paralog or homolog of Oryza sativa ERF922, represented by the protein sequence of SEQ ID NO:36 or the coding sequence / cDNA sequence of SEQ ID NO:51.

[0239] In certain embodiments, a plant (or plant part or population) contains more than one ERF922 gene, which can be homologous genes or paralogous genes, or both. In certain embodiments, a plant (or plant part or population) contains two or more ERF922 genes, which can be homologous genes or paralogous genes, or both.

[0240] In certain embodiments, different ERF922 homologs or paralogs are differentially expressed. Differential expression may require expression in different cells, tissues or organs; expression at different growth stages; or both.

[0241] In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, the CPL1 and / or ERF922 genes are mutated.

[0242] In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, the CPL1 and ERF922 genes are mutated.

[0243] In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, the CPL1 gene is mutated.

[0244] In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, the ERF922 gene is mutated.

[0245] In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, at least two, such as two, CPL1 and / or at least two, such as two, ERF922 genes are mutated. In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, at least two, such as two, CPL1 and at least two, such as two, ERF922 genes are mutated. In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, at least two, such as two, CPL1 genes are mutated. In certain embodiments of the methods, plants (or parts or populations thereof), uses, polynucleic acids or polypeptides according to the invention as described herein, at least two, such as two, ERF922 genes are mutated.

[0246] The skilled person will understand that a wild-type or unmutated gene product is a functional gene product with (substantially) unaltered functionality, such as the enzymatic or transcriptional activity as defined elsewhere herein. The skilled person will further understand that the sequence variations of the above wild-type genes do not include frameshift or nonsense mutations.

[0247] As used herein, a mutated CPL1 and / or ERF922 or a mutation in CPL1 and / or ERF922 can include or can refer to any type of CPL1 and / or ERF922 mutation. In certain embodiments, the mutation alters the expression of wild-type or native CPL1 and / or ERF922 protein and / or mRNA. In certain embodiments, the mutation reduces or eliminates (wild-type or native) CPL1 and / or ERF922 protein and / or mRNA expression, as described elsewhere herein. The mutation can affect transcription and / or translation. The mutation can occur in an exon or an intron. The mutation can occur in regulatory elements such as promoters, enhancers, terminators, insulators, etc., as well as in the 5' and / or 3' UTR coding regions. The mutation can occur in the coding sequence. The mutation can occur at a splice signal site such as a splice donor or acceptor site. The mutation can be a frameshift mutation. The mutation can be a nonsense mutation. The mutation can be a point mutation. The mutation can be an insertion or deletion of one or more nucleotides internally and / or terminally. The mutation can be a non-conservative mutation (where one or more wild-type amino acids are replaced by one or more non-wild-type amino acids). The mutation can result in a truncated protein. The mutation can affect or alter the function of the CPL1 and / or ERF922 protein, such as enzymatic activity or transcriptional activity. The mutation can reduce or (substantially) eliminate the function of the CPL1 and / or ERF922 protein, such as enzymatic activity or transcriptional activity. A reduced function such as reduced enzymatic activity or transcriptional activity can refer to a reduction of at least about 10%, preferably at least 30%, more preferably at least 50%, such as at least 20%, 40%, 60%, 80% or more, such as at least 85%, at least 90%, at least 95% or more. A (substantially) eliminated function, such as a (substantially) eliminated enzymatic activity or transcriptional activity, can refer to a reduction of at least 80%, preferably at least 90%, more preferably at least 95%. The mutation can be a dominant negative mutation.

[0248] In certain embodiments, only a single allele is mutated (e.g., a plant, plant part, or plant population contains only one mutant allele). In certain embodiments, multiple alleles are mutated. In certain embodiments, the mutation is homozygous. In certain embodiments, the mutation is heterozygous. In certain embodiments, one or two alleles of one or more paralogous genes are mutated. In certain embodiments, one or two alleles of one or more orthologous genes are mutated. In certain embodiments, one or two alleles of one or more paralogous and orthologous genes are mutated.

[0249] In certain embodiments, dominant negative CPL1 and / or ERF922 are expressed in plants. In certain embodiments, dominant negative CPL1 and / or ERF922 are expressed in plants, whereby the dominant negative is from the same species as the species in which it is expressed. In certain embodiments, dominant negative CPL1 and / or ERF922 are expressed in plants such that the dominant negative is from a different species than the species in which it is expressed (i.e., from an ortholog).

[0250] In certain embodiments, the endogenous CPL1 and / or ERF922 genes are mutated to result in the expression of a dominant negative protein. If all alleles are mutated (all homologous or paralogous genes), this dominant negative can be functionally equivalent to a knockout. If only a subset of alleles are mutated, this dominant negative may be functionally equivalent to a knockdown. In certain embodiments, the dominant negative is homozygous. In certain embodiments, the dominant negative is heterozygous.

[0251] In certain embodiments, dominant negative CPL1 and / or ERF922 are exogenous (i.e., recombinant or transgenic), although it may be derived from the same species as the species into which it is introduced.

[0252] As used herein, the term "CPL1 activity" can refer to the enzymatic activity of CPL1 or the non-enzymatic activity of CPL1. The term "having reduced CPL1 activity" in the context of variant CPL1 as described above in certain preferred embodiments refers to CPL1 whose enzymatic or non-enzymatic activity is affected, particularly CPL1 having reduced activity compared to wild-type or native CPL1. In certain embodiments, the (enzymatic) activity is at most 50% of wild-type CPL1 activity, preferably at most 40%, more preferably at most 30%, even more preferably at most 20%, most preferably at most 10%, for example at most 5%. The (enzymatic) activity can be measured by methods known in the art.

[0253] As used herein, the term "ERF922 activity" can refer to the transcriptional activity of ERF922 or the non-transcriptional activity of ERF922. The term "having reduced ERF922 activity" in the context of variant ERF922 as described above in certain preferred embodiments refers to ERF922 whose transcriptional or non-transcriptional activity is affected, particularly ERF922 having reduced activity compared to wild-type or native ERF922. In certain embodiments, the (transcriptional) activity is at most 50% of wild-type ERF922 activity, preferably at most 40%, more preferably at most 30%, even more preferably at most 20%, most preferably at most 10%, for example at most 5%. The (transcriptional) activity can be measured by methods known in the art.

[0254] In certain embodiments, the CPL1 and / or ERF922 mutation is the insertion of one or more nucleotides in the coding sequence. In certain embodiments, the CPL1 and / or ERF922 mutation is a nonsense mutation. In certain embodiments, the CPL1 and / or ERF922 mutation results in decreased expression of the CPL1 and / or ERF922 gene. In certain embodiments, the CPL1 and / or ERF922 mutation results in knockout of the CPL1 and / or ERF922 gene or knockdown of the CPL1 and / or ERF922 mRNA and / or protein. In certain embodiments, the mutation results in a frameshift in the coding sequence of CPL1 and / or ERF922. In certain embodiments, the mutation results in an alteration of the protein sequence encoded by the CPL1 and / or ERF922 gene.

[0255] In certain embodiments, the CPL1 and / or ERF922 mutation is the insertion of one or more nucleotides, preferably in an exon, preferably in the first exon, and preferably a frameshift insertion.

[0256] CPL1 and / or ERF922 mRNA and / or protein expression can be reduced or eliminated by mutating the CPL1 and / or ERF922 gene itself (including coding, non-coding, and regulatory elements). Methods for introducing mutations are described elsewhere herein. Alternatively, CPL1 and / or ERF922 mRNA and / or protein expression can be reduced or eliminated by (specifically) interfering with transcription and / or translation, such as reducing or eliminating mRNA and / or protein transcription or translation. Alternatively, CPL1 and / or ERF922 mRNA and / or protein expression can be reduced or eliminated by (specifically) interfering with mRNA and / or protein stability, such as reducing mRNA and / or protein stability. For example, mRNA (stability) can be reduced by RNAi as described elsewhere herein. miRNA can also be used to affect mRNA (stability). In certain embodiments, reduced CPL1 and / or ERF922 expression achieved by reducing mRNA or protein stability is also included in the term "mutated" CPL1 and / or ERF922. In certain embodiments, reduced CPL1 and / or ERF922 expression achieved by reducing mRNA or protein stability is not included in the term "mutated" CPL1 and / or ERF922.

[0257] As used herein, "mutation" refers to modifications at the DNA level, including genetic and / or epigenetic changes. Genetic alterations can include insertions, deletions, introduction of stop codons, base changes (e.g., transitions or transversions), or changes in splice junctions. These alterations may occur in the coding or non-coding regions of the endogenous DNA sequence (e.g., promoter regions, exons, introns, or splice junctions). For example, a genetic alteration can be a substitution of at least one nucleobase in the endogenous DNA sequence or in the regulatory sequence of the endogenous DNA sequence. For example, if such a nucleobase substitution occurs in a promoter, this can result in a change in the activity of the promoter because, for example, a cis-regulatory element is modified such that the affinity of a transcription factor for the mutated cis-regulatory element is altered compared to the wild-type promoter, and thus the activity of the promoter with the mutated cis-regulatory element is increased or decreased, depending on whether the transcription factor is a repressor or an inducer, or whether the affinity of the transcription factor for the mutated cis-regulatory element is enhanced or diminished. If, for example, such a nucleobase substitution occurs in the coding region of the endogenous DNA sequence, this can result in an amino acid substitution in the encoded protein, whereby the activity or stability of the protein can be altered compared to the wild-type protein. Epigenetic alterations can occur through changes in DNA methylation patterns.

[0258] Mutagenesis can be performed according to any technique known in the art. As used herein, "mutagenesis" or "performing mutagenesis" includes conventional mutagenesis and site-specific mutagenesis or "genome editing" or "gene editing". In conventional mutagenesis, modifications at the DNA level are not generated in a targeted manner. Plant cells or plants are exposed to mutagenic conditions by ultraviolet irradiation or using chemicals, such as TILLING (Till et al., 2004). Another method of random mutagenesis is mutagenesis by means of transposons. Site-specific mutagenesis enables the introduction of modifications at the DNA level at a predetermined position in the DNA in a targeted manner. For example, TALENS, meganucleases, homing endonucleases, zinc finger nucleases, or CRISPR / Cas systems, as further described herein, can be used for this purpose.

[0259] As used herein, the terms "introgress", "introgressing" and "introgressed" refer to both natural and artificial processes by which a chromosomal segment or gene of one species, variety or cultivar is introduced into the genome of another species, variety or cultivar by crossing species. This process can optionally be accomplished by backcrossing to a recurrent parent. For example, introgression of a desired allele at a particular locus can be passed to at least one offspring by sexual crossing between two parents of the same species, where at least one parent has the desired allele in its genome. Or, for example, transmission of the allele can occur by recombination between two donor genomes, such as in fused protoplasts, where at least one donor protoplast has the desired allele in its genome. The desired allele can be detected, for example, by markers associated with a phenotype, QTL, transgene, etc. In any case, the offspring containing the desired allele can be repeatedly backcrossed to a line with a desired genetic background and selected for the desired allele, resulting in fixation of the allele in the selected genetic background. When this process is repeated two or more times, the "introgression" process is generally referred to as "backcrossing". An "introgressed segment" or "introgressed section" or "introgressed region" refers to a chromosomal segment (or chromosomal part or region) that has been introduced into another plant of the same or a related species, either artificially or naturally, for example by hybridization or traditional breeding techniques such as backcrossing, i.e., the introgressed segment is the result of the breeding method (such as backcrossing) referred to by the verb "introgress". It should be understood that the term "introgressed segment" never includes an entire chromosome, but only a part of a chromosome. The introgressed segment can be large, for example even three-quarters or half of a chromosome, but is preferably smaller, for example about 15 Mb or less, for example about 10 Mb or less, about 9 Mb or less, about 8 Mb or less, about 7 Mb or less, about 6 Mb or less, about 5 Mb or less, about 4 Mb or less, about 3 Mb or less, about 2.5 Mb or 2 Mb or less, about 1 Mb (equal to 1,000,000 base pairs) or less, or about 0.5 Mb (equal to 500,000 base pairs) or less, for example about 200,000 bp (equal to 200 kilo base pairs) or less, about 100,000 bp (100 kb) or less, about 50,000 bp (50 kb) or less, about 25,000 bp (25 kb) or less.

[0260] The term "locus" (plural loci) refers to one or more specific locations or sites on a chromosome, where, for example, QTLs, genes, or genetic markers are found. As used herein, the term "quantitative trait locus" or "QTL" has its ordinary meaning known in the art. By way of further guidance, but not limited to, a QTL can refer to a DNA region associated with differential expression of a quantitative phenotypic trait in at least one genetic background, such as in at least one breeding population. The region of a QTL contains or is tightly linked to one or more genes that affect the trait in question. An "allele of a QTL" can comprise multiple genes or other genetic factors, such as a haplotype, within a contiguous genomic region or linkage group. An allele of a QTL can represent a haplotype within a specific window, where the window is a contiguous genomic region that can be defined and tracked using a set of one or more polymorphic markers. A haplotype can be defined by the unique fingerprint of the alleles at each marker within the specified window. A QTL can encode one or more alleles that affect the expression of a continuously distributed (quantitative) phenotype. In certain embodiments, a QTL as described herein can be homozygous. In certain embodiments, a QTL as described herein can be heterozygous.

[0261] As used herein, the term "allele" or "alleles" refers to one or more alternative forms of a locus, i.e., different nucleotide sequences.

[0262] As used herein, the term "mutant allele" or "mutation" of an allele includes an allele having one or more mutations, such as insertions, deletions, stop codons, base changes (e.g., transitions or transversions), or alterations of splice junctions, which may or may not result in an altered gene product. Modifications of an allele can occur in the coding or non-coding regions (e.g., promoter region, exon, intron, or splice junction).

[0263] As described elsewhere herein, a genetic element, introgression segment, or gene or allele that confers a trait (such as increased pathogen tolerance or resistance) is said to be "derivable from" or "obtainable from" or "derived from" or "present in" or "found in" a plant or plant part if it can be transferred from the plant in which it is present to another plant (such as a line or variety) in which it is not present using conventional breeding techniques that do not result in phenotypic changes in the recipient plant other than an increase in the trait conferred by the genetic element, locus, introgression segment, gene, or allele. These terms are used interchangeably, and thus a genetic element, locus, introgression segment, gene, or allele can be transferred into any other genetic background lacking that trait. Not only can plants containing the genetic element, locus, introgression segment, gene, or allele be used, but also the progeny / descendants of such plants that have been selected to retain the genetic element, locus, introgression segment, gene, or allele and are included herein. One or more techniques or combinations of techniques known in the art can be used by a person skilled in the art to determine whether a plant (or the genomic DNA, cells, or tissues of a plant) contains the same genetic element, locus, introgression segment, gene, or allele derivable from such a plant, such techniques being, for example, phenotypic assays, whole-genome sequencing, molecular marker analysis, trait mapping, chromosome painting, allele testing, etc. It should be understood that transgenic plants can also be included.

[0264] As used herein, the terms "genetic engineering", "transformation", and "genetic modification" are all used herein as synonyms for the transfer of isolated and cloned genes into the DNA of another organism, usually chromosomal DNA or the genome.

[0265] A "transgenic" or "genetically modified organism" (GMO) as used herein is an organism whose genetic material has been altered using techniques commonly referred to as "recombinant DNA technology". Recombinant DNA technology includes the ability to combine DNA molecules from different sources in vitro (such as in a test tube) into one molecule. This term generally does not include organisms whose genetic makeup has been altered by conventional hybridization breeding or "mutagenesis" breeding, as these methods predate the discovery of recombinant DNA technology. As used herein, "non-transgenic" refers to plants and foods derived from plants that are not "transgenic" or "genetically modified organisms" as defined above.

[0266] A "transgene" or "chimeric gene" refers to a locus containing a DNA sequence, such as a recombinant gene, that has been introduced into the genome of a plant by transformation, such as Agrobacterium-mediated transformation. A plant containing a transgene stably integrated into its genome is called a "transgenic plant".

[0267] "Gene editing" or "genome editing" refers to genetic engineering that inserts, deletes, modifies, or replaces DNA or RNA in the genome of a living organism. Gene editing can include targeted or non-targeted (random) mutagenesis. Targeted mutagenesis can be accomplished, for example, using engineered nucleases, such as using meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR / Cas9) systems. These nucleases create site-specific double-strand breaks (DSBs) at desired locations in the genome. The induced double-strand breaks are repaired by non-homologous end joining (NHEJ) or homologous recombination (HR), resulting in targeted mutations or nucleic acid modifications. The use of engineered nucleases is particularly suitable for generating gene knockouts or knockdowns. In certain embodiments, engineered nucleases that specifically induce mutations in the CPL1 and / or ERF922 genes have been developed, as described elsewhere herein, for example to generate mutant CPL1 and / or ERF922 or knockout the CPL1 and / or ERF922 genes. In certain embodiments, engineered nucleases that specifically target CPL1 and / or ERF922 mRNA have been developed, particularly RNA-specific CRISPR / Cas systems, for example to cleave CPL1 and / or ERF922 mRNA and generate knockdown of the CPL1 and / or ERF922 gene / mRNA / protein. Delivery and expression systems for engineered nuclease systems are well known in the art.

[0268] In some embodiments, the nuclease or targeted / site-specific / homing nuclease is, comprises, consists essentially of, or consists of: a (modified) CRISPR / Cas system or complex, a (modified) Cas protein, a (modified) zinc finger, a (modified) zinc finger nuclease (ZFN), a (modified) transcription activator-like effector (TALE), a (modified) transcription activator-like effector nuclease (TALEN), or a (modified) meganuclease. In some embodiments, the (modified) nuclease or targeted / site-specific / homing nuclease is, comprises, consists essentially of, or consists of a (modified) RNA-guided nuclease. It should be understood that in some embodiments, the nuclease may be codon-optimized for expression in plants. As used herein, the term "targeting" of a selected nucleic acid sequence means that the nuclease or nuclease complex acts in a nucleotide sequence-specific manner. For example, in the context of a CRISPR / Cas system, the guide RNA is capable of hybridizing to the selected nucleic acid sequence. As used herein, "hybridization" refers to a reaction in which one or more polynucleotides react to form a complex that is stabilized by hydrogen bonding between the bases of the nucleotide residues. The hydrogen bonding can occur through Watson Crick base pairing, Hoogstein binding, or in any other sequence-specific manner. The complex can comprise two strands forming a duplex structure, three or more strands forming a multi-stranded complex, a self-hybridizing strand, or any combination of these strands. Hybridization is the process by which a single-stranded nucleic acid molecule binds to a complementary nucleic acid strand, i.e., in accordance with such base pairing. For example, standard procedures for hybridization are described in Sambrook et al. (Molecular Cloning. A Laboratory Manual, Cold Spring Harbor Laboratory Press, 3rd edition 2001). Preferably, this will be understood to mean that at least 50%, more preferably at least 55%, 60%, 65%, 70%, 75%, 80%, or 85%, more preferably 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the nucleic acid strand bases form base pairs with the complementary nucleic acid strand. The hybridization reaction can form one step in a broader process, such as the initiation of PGR, or the cleavage of a polynucleotide by an enzyme. A sequence capable of hybridizing to a given sequence is called the "complement" of the given sequence.

[0269] Gene editing can involve transient, inducible, or constitutive expression of a gene editing component or system. Gene editing can involve genomic integration or episomal presence of a gene editing component or system. The gene editing component or system can be provided on a vector, such as a plasmid, which can be delivered by a suitable delivery vehicle, as known in the art. A preferred vector is an expression vector.

[0270] Gene editing can include providing a recombination template to effect homology-directed repair (HDR). For example, a genetic element can be replaced by gene editing that provides a recombination template. DNA can be cleaved upstream and downstream of the sequence to be replaced. Thereby, the sequence to be replaced is excised from the DNA. By HDR, the excised sequence is subsequently replaced by the template. In certain embodiments, the QTL alleles of the invention as described herein can be provided on / serve as the template. By designing the system such that double-strand breaks are introduced upstream and downstream of the corresponding region in the plant genome that does not contain the QTL allele, that region is excised and can be replaced by a template containing the QTL allele of the invention. In this way, introducing the QTL alleles of the invention into a plant does not require multiple backcrosses, especially in plants with a specific genetic background. Similarly, the mutant CPL1 and / or ERF922 of the invention can be provided on / serve as the template. However, more advantageously, the mutant CPL1 and / or ERF922 of the invention can be generated without using a recombination template, but only by endonuclease action resulting in a double-strand DNA break that is repaired by NHEJ, resulting in the generation of indels.

[0271] In certain embodiments, nucleic acid modification or mutation is affected by a (modified) transcription activator-like effector nuclease (TALEN) system. Transcription activator-like effectors (TALEs) can be engineered to bind to almost any desired DNA sequence. Exemplary methods for genome editing using the TALEN system can be found, for example, in Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, et al. Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res. 2011;39:e82; Zhang F, Cong L, Lodato S, Kosuri S, Church GM, Arlotta P. Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription. Nat Biotechnol. 2011;29:149–153 and U.S. Patent Nos. 8,450,471, 8,440,431, and 8,440,432, all of which are hereby specifically incorporated by reference. By way of further guidance, but not limited to, a naturally occurring TALE or “wild-type TALE” is a nucleic acid-binding protein secreted by various proteobacteria. A TALE polypeptide contains a nucleic acid-binding domain composed of tandem repeats of highly conserved monomeric polypeptides, the monomeric polypeptides being predominantly 33, 34, or 35 amino acids in length and differing from each other predominantly at amino acid positions 12 and 13. In advantageous embodiments, the nucleic acid is DNA. As used herein, the term “polypeptide monomer” or “TALE monomer” will be used to refer to the highly conserved repeat polypeptide sequences within the TALE nucleic acid-binding domain, and the term “repeat variable diresidue” or “RVD” will be used to refer to the highly variable amino acids at positions 12 and 13 of the polypeptide monomer. As provided throughout the disclosure, the amino acid residues of the RVD are described using the IUPAC single-letter amino acid code. The general representation of a TALE monomer contained within the DNA-binding domain is X1-11-(X12X13)-X14-33 or 34 or 35, where the subscripts denote amino acid positions and X denotes any amino acid. X12X13 represents the RVD. In some polypeptide monomers, the variable amino acid at position 13 is absent or non-existent, and in such polypeptide monomers, the RVD consists of a single amino acid. In this case, the RVD can alternatively be represented as X*, where X represents X12 and (*) represents the absence of X13.The DNA binding domain contains multiple TALE monomer repeats, which can be represented as (X1-11-(X12X13)-X14-33 or 34 or 35)z, where in one advantageous embodiment, z is at least 5 to 40. In another advantageous embodiment, z is at least 10 to 26. The TALE monomer has nucleotide binding affinity, which is determined by the identity of the amino acids in its RVD. For example, a polypeptide monomer with an RVD of NI preferentially binds adenine (A), a polypeptide monomer with an RVD of NG preferentially binds thymine (T), a polypeptide monomer with an RVD of HD preferentially binds cytosine (C), and a polypeptide monomer with an RVD of NN preferentially binds adenine (A) and guanine (G). In another embodiment of the present invention, a polypeptide monomer with an RVD of IG preferentially binds T. Thus, the number and order of the polypeptide monomer repeats in the nucleic acid binding domain of TALE determine its nucleic acid target specificity. In a further embodiment of the present invention, a polypeptide monomer with an RVD of NS recognizes all four base pairs and can bind A, T, G, or C. The structure and function of TALE are further described in, for example, Moscou et al., Science 326:1501 (2009), Boch et al., Science 326:1509-1512 (2009), and Zhang et al., Nature Biotechnology 29:149-153 (2011), each of which is incorporated herein by reference in its entirety.

[0272] In certain embodiments, nucleic acid modification or mutation is achieved by a (modified) zinc finger nuclease (ZFN) system. The ZFN system uses an artificial restriction enzyme generated by fusing a zinc finger DNA-binding domain to a DNA cleavage domain, which can be engineered to target a desired DNA sequence. Example methods of genome editing using ZFNs can be found in U.S. Patent Nos. 6,534,261, 6,607,882, 6,746,838, 6,794,136, 6,824,978, 6,866,997, 6,933,113, 6,979,539, 7,013,219, 7,030,215, 7,220,719, 7,241,573, 7,241,574, 7,585,849, 7,595,376, 6,903,185, and 6,479,626, all of which are hereby specifically incorporated by reference. By way of further guidance, but not limited to, artificial zinc finger (ZF) technology involves arrays of ZF modules to target new DNA-binding sites in the genome. Each finger module in the ZF array targets three DNA bases. Arrays of custom-designed individual zinc finger domains are assembled into zinc finger proteins (ZFPs). A ZFP can contain a functional domain. The first synthetic zinc finger nuclease (ZFN) was developed by fusing a ZF protein to the catalytic domain of the type IIS restriction enzyme FokI (Kim, Y.G. et al., 1994, Chimeric restriction endonuclease, Proc. Natl. Acad. Sci. U.S.A. 91, 883-887; Kim, Y.G. et al., 1996, Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain. Proc. Natl. Acad. Sci. U.S.A. 93, 1156-1160). By using paired ZFN heterodimers, cleavage specificity can be enhanced by reducing off-target activity, with each heterodimer targeting different nucleotide sequences separated by a short spacer (Doyon, Y. et al., 2011, Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures. Nat. Methods 8, 74-79). ZFPs can also be designed to act as transcriptional activators and repressors and have been used to target many genes in a variety of organisms.

[0273] In certain embodiments, the nucleic acid modification is achieved by (modified) meganucleases, which are endodeoxyribonucleases characterized by large recognition sites (double-stranded DNA sequences of 12 to 40 base pairs). Exemplary methods of using meganucleases can be found in U.S. Patent Nos.: 8,163,514; 8,133,697; 8,021,867; 8,119,361; 8,119,381; 8,124,369 and 8,129,134, which are hereby specifically incorporated by reference.

[0274] In certain embodiments, the nucleic acid modification is achieved by a (modified) CRISPR / Cas complex or system. For general information regarding CRISPR / Cas systems, their components, and the delivery of such components, including methods, materials, delivery vehicles, vectors, particles, and their manufacture and use, including amounts and formulations, as well as eukaryotic cells expressing Cas9 CRISPR / Cas, eukaryotes expressing Cas-9 CRISPR / Cas, reference is made to the following patent documents: U.S. Patent Nos. 8,999,641, 8,993,233, 8,697,359, 8,771,945, 8,795,965, 8,865,406, 8,871,445, 8,889,356, 8,889,418, 8,895,308, 8,906,616, 8,932,814, 8,945,839, 8,993,233, and 8,999,641; U.S. Patent Publications US2014-0310830 (US App. Ser. No. 14 / 105,031), US 2014-0287938 A1 (U.S. App. Ser. No. 14 / 213,991), US 2014-0273234 A1 (U.S. App. Ser. No. 14 / 293,674), US2014-0273232A1 (U.S. App. Ser. No. 14 / 290,575), US 2014-0273231 (U.S. App. Ser. No. 14 / 259,420), US2014-0256046 A1 (U.S. App. Ser. No. 14 / 226,274), US 2014-0248702 A1 (U.S. App. Ser. No. 14 / 258,458), US 2014-0242700 A1 (U.S. App. Ser. No. 14 / 222,930), US2014-0242699A1 (U.S. App. Ser. No. 14 / 183,512), US 2014-0242664 A1 (U.S. App. Ser. No. 14 / 104,990), US 2014-0234972 A1 (U.S. App. Ser. No. 14 / 183,471), US2014-0227787 A1 (U.S. App. Ser. No. 14 / 256,912), US 2014-0189896 A1 (U.S. App. Ser. No. 14 / 105,035), US2014-0186958 (U.S. App. Ser. No. 14 / 105,017), US2014-0186919 A1 (U.S. App. Ser. No. 14 / 104,977),US 2014-0186843 A1 (U.S. App. Ser. No. 14 / 104,900), US 2014-0179770 A1 (U.S. App. Ser. No. 14 / 104,837), and US 2014-0179006 A1 (U.S. App. Ser. No. 14 / 183,486), US 2014-0170753 (US App Ser No 14 / 183,429); US 2015-0184139 (U.S. App. Ser. No. 14 / 324,960); 14 / 054,414 European patent applications EP2 771 468 (EP13818570.7), EP 2 764 103 (EP13824232.6), and EP 2 784 162 (EP14170383.5); and PCT patent publications WO 2014 / 093661 (PCT / US2013 / 074743), WO 2014 / 093694 (PCT / US2013 / 074790), WO 2014 / 093595 (PCT / US2013 / 074611), WO 2014 / 093718 (PCT / US2013 / 074825), WO 2014 / 093709 (PCT / US2013 / 074812), WO 2014 / 093622 (PCT / US2013 / 074667), WO 2014 / 093635 (PCT / US2013 / 074691), WO 2014 / 093655 (PCT / US2013 / 074736), WO 2014 / 093712 (PCT / US2013 / 074819), WO 2014 / 093701 (PCT / US2013 / 074800), WO 2014 / 018423 (PCT / US2013 / 051418), WO 2014 / 204723 (PCT / US2014 / 041790), WO 2014 / 204724 (PCT / US2014 / 041800), WO 2014 / 204725 (PCT / US2014 / 041803), WO 2014 / 204726 (PCT / US2014 / 041804), WO 2014 / 204727 (PCT / US2014 / 041806), WO 2014 / 204728 (PCT / US2014 / 041808), WO 2014 / 204729 (PCT / US2014 / 041809), WO 2015 / 089351 (PCT / US2014 / 069897),WO 2015 / 089354 (PCT / US2014 / 069902), WO 2015 / 089364 (PCT / US2014 / 069925), WO 2015 / 089427 (PCT / US2014 / 070068), WO 2015 / 089462 (PCT / US2014 / 070127), WO 2015 / 089419 (PCT / US2014 / 070057), WO 2015 / 089465 (PCT / US2014 / 070135), WO 2015 / 089486 (PCT / US2014 / 070175), PCT / US2015 / 051691, PCT / US2015 / 051830. Also refer to U.S. Provisional Patent Applications 61 / 758,466, 61 / 802,174, 61 / 806,375, 61 / 814,263, 61 / 819,803, and 61 / 828,130 filed on January 30, 2013, March 15, 2013, March 5, 2013, May 28, 2013, respectively. Also refer to U.S. Provisional Patent Application 61 / 836,123 filed on June 17, 2013. Additionally refer to U.S. Provisional Patent Applications 61 / 835,931, 61 / 835,936, 61 / 835,973, 61 / 836,080, 61 / 836,101, and 61 / 836,127, all filed on June 17, 2013. Further refer to U.S. Provisional Patent Applications 61 / 862,468 and 61 / 862,355 filed on August 5, 2013; 61 / 871,301 filed on August 28, 2013; 61 / 960,777 filed on September 25, 2013, and 61 / 961,980 filed on October 28, 2013. Further refer to: PCT / US2014 / 62558 filed on October 28, 2014, and U.S. Provisional Patent Application Series 61 / 915,148, 61 / 915,150, 61 / 915,153, 61 / 915,203, 61 / 915,251, 61 / 915,301, 61 / 915,267, 61 / 915,260, and 61 / 915,397, all filed on December 12, 2013; 61 / 757,972 and 61 / 768,959 filed on January 29, 2013 and February 25, 2013, respectively; 62 / 010,888 and 62 / 010,879 both filed on June 11, 2014; 62 / 010,329 filed on June 10, 2014,62 / 010,439 and 62 / 010,441; 61 / 939,228 and 61 / 939,242, both filed on Feb. 12, 2014; 61 / 980,012 filed on Apr. 15, 2014; 62 / 038,358 filed on Aug. 17, 2014; 62 / 055,484, 62 / 055,460 and 62 / 055,487, all filed on Sep. 25, 2014, and 62 / 069,243 filed on Oct. 27, 2014. Reference is made to PCT Application PCT / US14 / 41806, filed on Jun. 10, 2014 and designating the United States. Reference is made to U.S. Provisional Patent Application 61 / 930,214, filed on Jan. 22, 2014. Reference is made to PCT Application PCT / US14 / 41806, filed on Jun. 10, 2014 and designating the United States. Also mentioned are U.S. Application 62 / 180,709, 17-Jun-15, PROTECTED GUIDE RNAS (PGRNAS); U.S. Application 62 / 091,455, filed on Dec. 12, 2014, PROTECTED GUIDE RNAS (PGRNAS); U.S. Application 62 / 096,708, 24-Dec-14, PROTECTED GUIDE RNAS (PGRNAS); U.S. Application 62 / 091,462, 12-Dec-14, 62 / 096,324, 23-Dec-14, 62 / 180,681, 17-Jun-2015, and 62 / 237,496, 5-Oct-2015, DEAD GUIDES FOR CRISPR TRANSCRIPTION FACTORS; U.S. Applications 62 / 091,456, 12-Dec-14 and 62 / 180,692, 17-Jun-2015, ESCORTED AND FUNCTIONALIZED GUIDES FOR CRISPR-CAS SYSTEMS; U.S. Application 62 / 091,461, 12-Dec-14, DELIVERY, USE AND THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS AND COMPOSITIONS FOR GENOME EDITING AS TO HEMATOPOETIC STEM CELLS (HSCs); U.S. Application 62 / 094,903, 19-Dec-14,UNBIASED IDENTIFICATION OF DOUBLE-STRAND BREAKS AND GENOMIC REARRANGEMENT BY GENOME-WISE INSERT CAPTURE SEQUENCING; US Application 62 / 096,761, 24-Dec-14, ENGINEERING OF SYSTEMS, METHODS AND OPTIMIZED ENZYME AND GUIDE SCAFFOLDS FOR SEQUENCE MANIPULATION; US Application 62 / 098,059, 30-Dec-14, 62 / 181,641, 18-Jun-2015, and 62 / 181,667, 18-Jun-2015, RNA-TARGETING SYSTEM; US Applications 62 / 096,656, 24-Dec-14 and 62 / 181,151, 17-Jun-2015, CRISPR HAVING OR ASSOCIATED WITH DESTABILIZATION DOMAINS; US Application 62 / 096,697, 24-Dec-14, CRISPR HAVING OR ASSOCIATED WITH AAV; US Application 62 / 098,158, 30-Dec-14, ENGINEERED CRISPR COMPLEX INSERTIONAL TARGETING SYSTEMS; US Application 62 / 151,052, 22-Apr-15, CELLULAR TARGETING FOR EXTRACELLULAR EXOSOMAL REPORTING; US Application 62 / 054,490, 24-Sep-14, DELIVERY, USE AND THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS AND COMPOSITIONS FOR TARGETING DISORDERS AND DISEASES USING PARTICLE DELIVERY COMPONENTS; US Application 61 / 939,154, 12-FEB-14, SYSTEMS, METHODS AND COMPOSITIONS FOR SEQUENCE MANIPULATION WITH OPTIMIZED FUNCTIONAL CRISPR-CAS SYSTEMS; US Application 62 / 055,484, 25-Sep-14, SYSTEMS,METHODS AND COMPOSITIONS FOR SEQUENCE MANIPULATION WITH OPTIMIZED FUNCTIONAL CRISPR-CAS SYSTEMS; US Application 62 / 087,537, 4-Dec-14, SYSTEMS, METHODS AND COMPOSITIONS FOR SEQUENCE MANIPULATION WITH OPTIMIZED FUNCTIONAL CRISPR-CAS SYSTEMS; US Application 62 / 054,651, 24-Sep-14, DELIVERY, USE AND THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS and COMPOSITIONS FOR MODELING COMPETITION OF MULTIPLE CANCER MUTATIONS IN VIVO; US Application 62 / 067,886, 23-Oct-14, DELIVERY, USE AND THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS AND COMPOSITIONS FOR MODELING COMPETITION OF MULTIPLE CANCER MUTATIONS IN VIVO; US Applications 62 / 054,675, 24-Sep-14 and 62 / 181,002, 17-Jun-2015, DELIVERY, USE AND THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS and COMPOSITIONS IN NEURONAL CELLS / TISSUES; US Application 62 / 054,528, 24-Sep-14, DELIVERY, USE AND THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS AND COMPOSITIONS IN IMMUNE DISEASES OR DISORDERS; US Application 62 / 055,454, 25-Sep-14, DELIVERY,USE and THERAPEUTIC APPLICATIONS OF THE CRISPR-CAS SYSTEMS AND COMPOSITIONS FOR TARGETING DISORDERS and DISEASES USING CELL PENETRATION PEPTIDES (CPP); US Application 62 / 055,460, 25-Sep-14, MULTIFUNCTIONAL-CRISPR COMPLEXES AND / OR OPTIMIZED ENZYME LINKED FUNCTIONAL-CRISPR COMPLEXES; US Application 62 / 087,475, 4-Dec-14 and 62 / 181,690, 18-Jun-2015, FUNCTIONAL SCREENING WITH OPTIMIZED FUNCTIONAL CRISPR-CAS SYSTEMS; US Application 62 / 055,487, 25-Sep-14, FUNCTIONAL SCREENING WITH OPTIMIZED FUNCTIONAL CRISPR-CAS SYSTEMS; US Application 62 / 087,546, 4-Dec-14 and 62 / 181,687, 18-Jun-2015, MULTIFUNCTIONAL CRISPR COMPLEXES AND / OR OPTIMIZED ENZYME LINKED FUNCTIONAL-CRISPR COMPLEXES; and US Application 62 / 098,285, 30-Dec-14, CRISPR MEDIATED IN VIVO MODELING AND GENETICS SCREENING OF TUMOR GROWTH and METASTASIS. Mentioned US Applications 62 / 181,659, 18-Jun-2015 and 62 / 207,318, 19-Aug-2015, ENGINEERING AND OPTIMIZATION OF SYSTEMS, METHODS, ENZYME AND GUIDE SCAFFOLDS OF CAS9 ORTHOLOGS AND VARIANTS FOR SEQUENCE MANIPULATION. Mentioned US Applications 62 / 181,663, 18-Jun-2015 and 62 / 245,264, 22-Oct-2015, NOVEL CRISPR ENZYMES AND SYSTEMS, US Application 62 / 181,675, 18-Jun-2015,and Attorney Docket No. 46783.01.2128, filed Oct 22, 2015, NOVEL CRISPR ENZYMES AND SYSTEMS, U.S. Application No. 62 / 232,067, filed Sep 24, 2015, U.S. Application No. 62 / 205,733, filed Aug 16, 2015, U.S. Application No. 62 / 201,542, filed Aug 5, 2015, U.S. Application No. 62 / 193,507, filed Jul 16, 2015, and U.S. Application No. 62 / 181,739, filed Jun 18, 2015, all with the title NOVEL CRISPR ENZYMES AND SYSTEMS and U.S. Application No. 62 / 245,270, filed Oct 22, 2015, NOVEL CRISPR ENZYMES AND SYSTEMS. Also mentioned are U.S. Application No. 61 / 939,256, filed Feb 12, 2014, and WO 2015 / 089473 (PCT / US2014 / 070152), filed Dec 12, 2014, both with the title ENGINEERING OF SYSTEMS, METHODS AND OPTIMIZED GUIDE COMPOSITIONS WITH NEW ARCHITECTURES FOR SEQUENCE MANIPULATION. Also mentioned are PCT / US2015 / 045504, filed Aug 15, 2015, U.S. Application No. 62 / 180,699, filed Jun 17, 2015, and U.S. Application No. 62 / 038,358, filed Aug 17, 2014, all with the title GENOME EDITING USING CAS9 NICKASES. European Patent Application EP3009511. Further incorporated is Multiplex genome engineering using CRISPR / Cas systems. Cong, L., Ran, F.A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P.D., Wu, X., Jiang, W., Marraffini, L.A., & Zhang, F. Science Feb 15; 339(6121):819-23 (2013); RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Jiang W., Bikard D., Cox D., Zhang F,Marraffini LA. Nat Biotechnol Mar; 31(3):233 - 9(2013); One - Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR / Cas - Mediated Genome Engineering. Wang H., Yang H., Shivalila CS., Dawlaty MM., Cheng AW., Zhang F., Jaenisch R. Cell May 9; 153(4):910 - 8(2013); Optical control of mammalian endogenous transcription and epigenetic states. Konermann S, Brigham MD, Trevino AE, Hsu PD, Heidenreich M, Cong L, Platt RJ, Scott DA, Church GM, Zhang F. Nature. 2013 Aug 22; 500(7463):472 - 6. doi:10.1038 / Nature12466. Epub 2013 Aug 23; Double Nicking by RNA - Guided CRISPR Cas9 for Enhanced Genome Editing Specificity. Ran, FA., Hsu, PD., Lin, CY., Gootenberg, JS., Konermann, S., Trevino, AE., Scott, DA., Inoue, A., Matoba, S., Zhang, Y., & Zhang, F. Cell Aug 28. pii:S0092 - 8674(13)01015 - 5.(2013); DNA targeting specificity of RNA - guided Cas9 nucleases. Hsu, P., Scott, D., Weinstein, J., Ran, FA., Konermann, S., Agarwala, V., Li, Y., Fine, E., Wu, X., Shalem, O., Cradick, TJ., Marraffini, LA., Bao, G., & Zhang,F.Nat Biotechnol doi:10.1038 / nbt.2647(2013);Genome engineering usingthe CRISPR-Cas9system.Ran,FA.,Hsu,PD.,Wright,J.,Agarwala,V.,Scott,DA.,Zhang,F.Nature Protocols Nov;8(11):2281-308.(2013);Genome-Scale CRISPR-Cas9Knockout Screening in Human Cells.Shalem,O.,Sanjana,NE.,Hartenian,E.,Shi,X.,Scott,DA.,Mikkelson,T.,Heckl,D.,Ebert,BL.,Root,DE.,Doench,JG.,Zhang,F.ScienceDec 12.(2013).[Epub ahead of print];Crystal structure of cas9 in complex withguide RNA and target DNA.Nishimasu,H.,Ran,FA.,Hsu,PD.,Konermann,S.,Shehata,SI.,Dohmae,N.,Ishitani,R.,Zhang,F.,Nureki,O.Cell Feb 27.(2014).156(5):935-49;Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells.Wu X.,Scott DA.,Kriz AJ.,Chiu AC.,Hsu PD.,Dadon DB.,Cheng AW.,Trevino AE.,KonermannS.,Chen S.,Jaenisch R.,Zhang F.,Sharp PA.Nat Biotechnol.(2014)Apr 20.doi:10.1038 / nbt.2889;CRISPR-Cas9Knockin Mice for Genome Editing和Cancer Modeling,Platt et al.,Cell 159(2):440 - 455(2014) DOI:10.1016 / j.cell.2014.09.014; Development and Applications of CRISPR-Cas9 for Genome Engineering, Hsu et al, Cell 157, 1262 - 1278(June 5, 2014)(Hsu 2014); Genetic screens in human cells using the CRISPR / Cas9 system, Wang et al., Science. 2014 January 3; 343(6166):80–84.doi:10.1126 / science.1246981; Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation, Doench et al., Nature Biotechnology 32(12):1262 - 7(2014) published online 3 September 2014; doi:10.1038 / nbt.3026, and In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9, Swiech et al, Nature Biotechnology 33, 102–106(2015) published online 19 October 2014; doi:10.1038 / nbt.3055, Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System, Zetsche et al., Cell 163, 1 - 13(2015); Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems, Shmakov et al., Mol Cell 60(3):385 - 397(2015); C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector,Abudayyeh et al., Science (2016), published online on June 2, 2016, doi:10.1126 / science.aaf5573. Each of these publications, patents, patent publications, and applications, as well as all documents cited therein or during their prosecution (“application cited documents”) and all documents cited or incorporated by reference in the application cited documents, together with any instructions, descriptions, product specifications, and product manuals of any products mentioned in or incorporated by reference in any of these documents, are hereby incorporated by reference into this application and may be used in the practice of the present invention. All documents (e.g., these patents, patent publications, and applications and the application cited documents) are incorporated by reference into this application to the same extent as if each individual document had been specifically and individually indicated to be incorporated by reference.,

[0275] In certain embodiments, the CRISPR / Cas system or complex is a type II CRISPR / Cas system. In certain embodiments, the CRISPR / Cas system or complex is a type II, V, or VI CRISPR / Cas system or complex. The CRISPR / Cas system does not require the generation of custom proteins that target specific sequences, but rather can program a single Cas protein to recognize a specific nucleic acid target via an RNA guide (gRNA). In other words, the Cas enzyme protein can be recruited to a specific nucleic acid target site of interest (which may consist of or be composed of RNA and / or DNA) using the short RNA guide.

[0276] Generally, as used in the foregoing documents herein, CRISPR / Cas or the CRISPR system collectively refers to transcripts and other elements involved in the expression or guidance of CRISPR-associated ("Cas") gene activity, including sequences encoding Cas genes and one or more tracr (trans-activating CRISPR) sequences (e.g., tracrRNA or the active portion of tracrRNA), tracr-mate sequences (including "direct repeats" and portions of direct repeats processed by tracrRNA in the context of endogenous CRISPR systems), guide sequences (also referred to as "spacers" in the context of endogenous CRISPR systems) or "RNA" as used herein (e.g., RNA guiding Cas such as Cas9, e.g., CRISPR RNA, and, where applicable, trans-activating (tracr) RNA or single guide RNA (sgRNA) (chimeric RNA)) or other sequences and transcripts from the CRISPR locus. Generally, the CRISPR system is characterized by elements that facilitate the formation of a CRISPR complex at a target sequence site (also referred to as a protospacer in the context of endogenous CRISPR systems). In the context of forming a CRISPR complex, a "target sequence" is a sequence to which a guide sequence is designed to have complementarity, wherein hybridization between the target sequence and the guide sequence facilitates the formation of the CRISPR complex. The target sequence can comprise any polynucleotide, e.g., DNA or RNA polynucleotide.

[0277] In certain embodiments, the gRNA is a chimeric guide RNA or single guide RNA (sgRNA). In certain embodiments, the gRNA comprises a guide sequence and a tracr mate sequence (or direct repeat). In certain embodiments, the gRNA comprises a guide sequence, a tracr mate sequence (or direct repeat) and a tracr sequence. In certain embodiments, the CRISPR / Cas system or complex as described herein does not comprise and / or is not dependent on the presence of a tracr sequence (e.g., if the Cas protein is Cpfl).

[0278] As used herein, the term "crRNA" or "guide RNA" or "single guide RNA" or "sgRNA" or "one or more nucleic acid components" of a CRISPR / Cas locus effector protein, as applicable, encompasses any polynucleotide sequence having sufficient complementarity to a target nucleic acid sequence to hybridize with the target nucleic acid sequence and direct sequence-specific binding of a nucleic acid targeting complex to the target nucleic acid sequence. In some embodiments, the degree of complementarity is about or greater than about 50%, 60%, 75%, 80%, 85%, 90%, 95%, 97.5%, 99% or more when optimal alignment is performed using a suitable alignment algorithm. Any suitable algorithm for aligning sequences can be used to determine the optimal alignment, non-limiting examples of which include the Smith-Waterman algorithm, the Needleman-Wunsch algorithm, algorithms based on the Burrows-Wheeler transform (e.g., Burrows Wheeler Aligner), ClustalW, Clustal X, BLAT, Novoalign (Novocraft Technologies; available at www.novocraft.com), ELAND (Illumina, San Diego, CA), SOAP (available at soap.genomics.org.cn) and Maq (available at maq.sourceforge.net). The ability of the guide sequence (within the guide RNA targeting the nucleic acid) to direct sequence-specific binding of the targeting nucleic acid complex to the target nucleic acid sequence can be evaluated by any suitable assay.

[0279] A guide sequence can be selected and thus a guide RNA targeting a nucleic acid can be selected to target any target nucleic acid sequence. The target sequence can be DNA. The target sequence can be genomic DNA. The target sequence can be mitochondrial DNA. The target sequence can be any RNA sequence. In some embodiments, the target sequence can be a sequence within an RNA molecule selected from the group consisting of: messenger RNA (mRNA), pre-mRNA, ribosomal RNA (rRNA), transfer RNA (tRNA), microRNA (miRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), double-stranded RNA (dsRNA), non-coding RNA (ncRNA), long non-coding RNA (lncRNA), and small cytoplasmic RNA (scRNA). In some preferred embodiments, the target sequence can be a sequence within an RNA molecule selected from mRNA, pre-mRNA, and rRNA. In some preferred embodiments, the target sequence can be a sequence within an RNA molecule selected from ncRNA and lncRNA. In some more preferred embodiments, the target sequence can be a sequence within an mRNA molecule or a pre-mRNA molecule.

[0280] In certain embodiments, the gRNA comprises a stem-loop, preferably a single stem-loop. In certain embodiments, the direct repeat forms a stem-loop, preferably a single stem-loop. In certain embodiments, the spacer region of the guide RNA is from 15 to 35 nt in length. In certain embodiments, the spacer region of the guide RNA is at least 15 nucleotides in length. In certain embodiments, the spacer region is from 15 to 17 nt, such as 15, 16 or 17 nt, 17 to 20 nt, such as 17, 18, 19 or 20 nt, 20 to 24 nt, such as 20, 21, 22, 23 or 24 nt, 23 to 25 nt, such as 23, 24 or 25 nt, 24 to 27 nt, such as 24, 25, 26 or 27 nt, 27 to 30 nt, such as 27, 28, 29 or 30 nt, 30 to 35 nt, such as 30, 31, 32, 33, 34 or 35 nt, or 35 nt or longer. In certain embodiments, the CRISPR / Cas system requires a tracrRNA. The "tracrRNA" sequence or a similar term includes any polynucleotide sequence having sufficient complementarity to hybridize with the crRNA sequence. In some embodiments, when optimally aligned, the degree of complementarity between the tracrRNA sequence and the crRNA sequence along the length of the shorter of the two sequences is about or greater than about 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97.5%, 99% or higher. In some embodiments, the tracr sequence is about or greater than about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50 or more nucleotides in length. In some embodiments, the tracr sequence and the gRNA sequence are included in a single transcript such that hybridization between the two results in a transcript having a secondary structure, such as a hairpin. In one embodiment of the invention, the transcript or the transcribed polynucleotide sequence has at least two or more hairpins. In a preferred embodiment, the transcript has two, three, four or five hairpins. In another embodiment of the invention, the transcript has at most five hairpins. In the hairpin structure, the last "N" of the loop and the 5' sequence portion upstream can correspond to the tracr mate sequence, and then the 3' sequence portion of the loop corresponds to the tracr sequence. In the hairpin structure, alternatively, the last "N" of the loop and the 5' sequence portion upstream can correspond to the tracr sequence, and the 3' sequence portion of the loop corresponds to the tracr mate sequence. In another embodiment, the CRISPR / Cas system does not require a tracrRNA, as known to those skilled in the art.

[0281] In some embodiments, the guide RNA (which can direct Cas to the target locus) can comprise (1) a guide sequence capable of hybridizing to the target locus, and (2) a tracr mate or direct repeat sequence (in the 5' to 3' direction, or 3' to 5' direction, as is known to those skilled in the art depending on the type of Cas protein). In certain embodiments, the CRISPR / Cas protein is characterized in that it utilizes a guide RNA comprising a guide sequence capable of hybridizing to the target locus and a direct repeat sequence, and does not require a tracrRNA. In certain embodiments, where the CRISPR / Cas protein is characterized in that it utilizes a tracrRNA, the guide sequence, tracr mate, and tracr sequence can be present in a single RNA, i.e., the sgRNA (arranged in the 5' to 3' direction or in the 3' to 5' direction), or the tracr RNA can be a different RNA from the RNA containing the guide and tracr mate sequences. In these embodiments, the tracr hybridizes to the tracr mate sequence and directs the CRISPR / Cas complex to the target sequence.

[0282] Generally, in the case of a system for endogenous targeting of nucleic acids, the formation of a complex targeting the nucleic acid (comprising a guide RNA that hybridizes to the target sequence and is complexed with one or more nucleic acid targeting effector proteins) results in the modification of one or both of the DNA or RNA strands in or near the target sequence (e.g., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50 or more base pairs of the target sequence). As used herein, the term "sequence associated with the target locus of interest" refers to a sequence adjacent to the target sequence (e.g., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50 or more base pairs of the target sequence, where the target sequence is contained within the target locus of interest). Those skilled in the art will appreciate that the selection is the specific cleavage site of the CRISPR / Cas system relative to the target sequence, as is known in the art, which can be within the target sequence or in the 3' or 5' sequence of the target sequence.

[0283] In some embodiments, the effector protein of the unmodified targeting nucleic acid can have nuclease activity. In some embodiments, the nuclease as described herein can direct cleavage of one or both nucleic acid (DNA, RNA, or hybrid, which can be single-stranded or double-stranded) strands at or near the position of the target sequence, e.g., within the target sequence and / or within the complementary sequence of the target sequence or at a sequence related to the target sequence. In some embodiments, the effector protein of the targeting nucleic acid can direct cleavage of one or both DNA or RNA strands within about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 100, 200, 500 or more base pairs from the first or last nucleotide of the target sequence. In some embodiments, the cleavage can be blunt-ended (e.g., for Cas9, such as SaCas9 or SpCas9). In some embodiments, the cleavage can be staggered (e.g., for Cpf1), i.e., generating sticky ends. In some embodiments, the cleavage is a staggered cleavage with a 5' overhang. In some embodiments, the cleavage is a staggered cleavage with a 5' overhang of 1 to 5 nucleotides, preferably 4 or 5 nucleotides. In some embodiments, the cleavage site is upstream of the PAM. In some embodiments, the cleavage site is downstream of the PAM. In some embodiments, the effector protein of the targeting nucleic acid can be mutated relative to the corresponding wild-type enzyme such that the mutated effector protein of the targeting nucleic acid lacks the ability to cleave one or both DNA or RNA strands of the target polynucleotide containing the target sequence. As another example, two or more catalytic domains of the Cas protein (e.g., RuvC I, RuvCII, and RuvC III or the HNH domain of the Cas9 protein) can be mutated to generate a mutated Cas protein that is substantially devoid of all DNA cleavage activity. In some embodiments, when the cleavage activity of the mutated enzyme is about no more than 25%, 10%, 5%, 1%, 0.1%, 0.01% or less of the nucleic acid cleavage activity of the non-mutated form of the enzyme, the effector protein of the targeting nucleic acid can be considered to be substantially devoid of all DNA and / or RNA cleavage activity; an example can be when the nucleic acid cleavage activity of the mutated form is zero or negligible compared to the non-mutated form. As used herein, the term "modified" Cas generally refers to a Cas protein having one or more modifications or mutations (including point mutations, truncations, insertions, deletions, chimeras, fusion proteins, etc.) compared to the wild-type Cas protein from which it is derived. Derived means that the enzyme derived therefrom is to a large extent based on having a high sequence homology with the wild-type enzyme, but it has been mutated (modified) in a manner known in the art or as described herein.

[0284] In some embodiments, the target sequence should be associated with a PAM (protospacer adjacent motif) or PFS (protospacer flanking sequence or site); that is, the PAM or PFS is a short sequence recognized by the CRISPR complex. The exact sequence and length requirements of the PAM vary depending on the CRISPR enzyme used, but the PAM is typically a 2- to 5-base pair sequence adjacent to the protospacer (i.e., the target sequence). Examples of PAM sequences are given in the Examples section below, and one of ordinary skill in the art will be able to identify additional PAM sequences for use with a given CRISPR enzyme. Additionally, engineering of the PAM-interacting (PI) domain can program PAM specificity, improve target site recognition fidelity, and increase the versatility of Cas, such as the Cas9 genome engineering platform. Cas proteins such as Cas9 protein can be engineered to alter their PAM specificities, as described, for example, in Kleinstiver BP et al. Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jul 23;523(7561):481-5. doi:10.1038 / nature14592. In some embodiments, the method comprises binding a CRISPR complex to a target polynucleotide to effect cleavage of the target polynucleotide, thereby modifying the target polynucleotide, wherein the CRISPR complex comprises a CRISPR enzyme complexed with a guide sequence that hybridizes to a target sequence within the target polynucleotide, wherein the guide sequence is linked to a tracr mate sequence that hybridizes to a tracr sequence. One of ordinary skill in the art will understand that other Cas proteins can be similarly modified.

[0285] As used herein, Cas proteins, such as but not limited to Cas9, Cpf1 (Cas12a), C2c1 (Cas12b), C2c2 (Cas13a), C2c3, Cas13b proteins, can be derived from any suitable source and thus can include different orthologs, derived from a variety of (prokaryotic) organisms, as is well documented in the art. In certain embodiments, the Cas protein is (modified) Cas9, preferably (modified) Staphylococcus aureus Cas9 (SaCas9) or (modified) Streptococcus pyogenes Cas9 (SpCas9). In certain embodiments, the Cas protein is (modified) Cpf1, preferably Acidaminococcus sp., such as Acidaminococcus BV3L6 Cpf1 (AsCpf1) or Lachnospiraceae bacterium Cpf1, such as Lachnospiraceae bacterium MA2020 or Lachnospiraceae bacterium MD2006 (LbCpf1). In certain embodiments, the Cas protein is (modified) C2c2, preferably Leptotrichia wadei C2c2 (LwC2c2) or Listeria newyorkensis FSL M6-0635 C2c2 (LbFSLC2c2). In certain embodiments, the (modified) Cas protein is C2c1. In certain embodiments, the (modified) Cas protein is C2c3. In certain embodiments, the (modified) Cas protein is Cas13b.

[0286] In some embodiments, nucleic acid modification is achieved by random mutagenesis. Cells or organisms can be exposed to mutagens, such as ultraviolet radiation or mutagenic chemicals (e.g., ethyl methanesulfonate (EMS)), and then mutants with desired characteristics are selected. For example, mutants can be identified by TILLING (Targeting Induced Local Lesions IN Genomes). This method combines mutagenesis, for example using a chemical mutagen such as ethyl methanesulfonate (EMS), with a sensitive DNA screening technique for identifying single-base mutations / point mutations in target genes. The TILLING method relies on the formation of DNA heteroduplexes, which form when multiple alleles are amplified by PCR and then heated and slowly cooled. "Bubbles" form at mismatches between the two DNA strands and are then cut by a single-strand nuclease. The products are then separated by size, for example by HPLC. See also McCallum et al. "Targeted screening for induced mutations"; Nat Biotechnol. 2000 Apr;18(4):455-7 and McCallum et al. "Targeting induced local lesions IN genomes (TILLING) for plant functional genomics"; Plant Physiol. 2000 Jun;123(2):439-42.

[0287] RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression or translation by neutralizing targeted mRNA molecules. Two types of small ribonucleic acid (RNA) molecules - microRNA (miRNA) and small interfering RNA (siRNA) - are crucial for RNA interference. RNA is a direct product of genes, and these small RNAs can bind to other specific messenger RNA (mRNA) molecules and increase or decrease their activity, for example, by preventing the mRNA from being translated into protein. The RNAi pathway exists in many eukaryotes, including animals, and is initiated by the Dicer enzyme, which cleaves long double-stranded RNA (dsRNA) molecules into short double-stranded fragments of siRNA (small interfering RNA) approximately 21 nucleotides in length. Each siRNA unfolds into two single-stranded RNAs (ssRNAs), the passenger strand and the guide strand. The passenger strand is degraded, and the guide strand is incorporated into the RNA-induced silencing complex (RISC). Mature miRNA is structurally similar to siRNA generated from exogenous dsRNA, but before maturation, miRNA must first undergo extensive post-transcriptional modification. miRNA is expressed from longer RNA-encoding genes as primary transcripts called pri-miRNA, which are processed in the nucleus by the microprocessor complex into a 70-nucleotide stem-loop structure called pre-miRNA. This complex consists of the Rnase III enzyme called Drosha and the dsRNA-binding protein DGCR8. The dsRNA portion of this pre-miRNA is bound and cleaved by Dicer, generating mature miRNA molecules that can be incorporated into the RISC complex; thus, miRNA and siRNA share the same downstream cellular machinery. Short hairpin RNA or small hairpin RNA (shRNA / hairpin vector) is an artificial RNA molecule with a tight hairpin turn and can be used to silence target gene expression by RNA interference. The best-studied outcome is post-transcriptional gene silencing, which occurs when the guide strand pairs with a complementary sequence in the messenger RNA molecule and induces cleavage by the catalytic component of RISC, Argonaute 2 (Ago2). As used herein, an RNAi molecule can be siRNA, shRNA, or miRNA. It should be understood that an RNAi molecule can be so applied to plants / plants or can be encoded by a suitable vector from which the RNAi molecule is expressed. Delivery and expression systems for RNAi molecules such as siRNA, shRNA, or miRNA are well known in the art.

[0288] In certain embodiments, methods for obtaining a plant or plant part according to the invention as described herein, such as methods for obtaining a plant or plant part having increased pathogen resistance or tolerance, involve or comprise genetic transformation and / or gene editing, such as including CRISPR / Cas, TALEN, ZFN, meganucleases; (induced) mutagenesis, which may or may not be random mutagenesis, such as TILLING. In certain embodiments, methods for obtaining a plant or plant part according to the invention as described herein, such as methods for obtaining a plant or plant part having increased pathogen resistance or tolerance, involve or comprise RNAi applications, which may or may not involve or comprise genetic transformation applications. By way of example, non-genetic transformation applications may include, for example, applying RNAi components such as double-stranded siRNA to the plant or plant surface, such as as a spray. Stable integration into the plant genome is not required.

[0289] In certain embodiments, methods for obtaining a plant or plant part according to the invention as described herein, such as methods for obtaining a plant or plant part having increased pathogen resistance or tolerance, do not involve or comprise genetic transformation, gene editing, and / or mutagenesis.

[0290] In certain embodiments, methods for obtaining a plant or plant part according to the invention as described herein, such as methods for obtaining a plant or plant part having increased pathogen resistance or tolerance, involve, comprise, consist of, or include breeding and selection.

[0291] In certain embodiments, methods for obtaining a plant or plant part according to the invention as described herein, such as methods for obtaining a plant or plant part having increased pathogen resistance or tolerance, do not involve, do not comprise, or do not consist of breeding and selection.

[0292] In one aspect, the invention relates to a plant or plant part obtainable or obtained by the method of the invention as described herein, such as a method for obtaining a plant or plant part having increased pathogen resistance or tolerance.

[0293] In certain embodiments of the methods, uses, plants, plant parts, plant populations, nucleic acids or proteins of the invention as described herein, CPL1 is mutated. In certain embodiments, the mutation is a dominant negative mutation. In certain embodiments, the mutation is located in the acyl phosphatase signature motif. In certain embodiments, the mutation is a mutation in the DXDXT motif, where D is aspartic acid, T is threonine, and X is any amino acid. For example, the DXDXT motif can be found at positions 128 to 134 of the Arabidopsis CPL4 protein. The corresponding DXDXT motif can be found at positions 161 to 165 of the Arabidopsis CPL1 protein, such as positions 161 to 165 of SEQ ID NO:1. Corresponding positions can be identified in orthologs, paralogs, homologs or xenologs. In certain embodiments, one or both of the aspartic acid residues are mutated. In certain embodiments, the threonine residue is mutated. In certain preferred embodiments, the first aspartic acid residue (i.e., the aspartic acid residue corresponding to position 161 of Arabidopsis CPL1) is mutated. In certain preferred embodiments, the mutation is a non-conservative mutation. In certain embodiments, the mutation is to a (small) non-polar amino acid. In certain embodiments, the mutation is to an alanine residue.

[0294] In certain embodiments, the CPL1 protein comprises one of the following mutations:

[0295] - D148X, preferably D148A, when the plant is from the genus Zea, preferably maize, preferably where the maize wild-type sequence has the sequence as shown in SEQ ID NO:2;

[0296] - D149X, preferably D149A, when the plant is from the genus Zea, preferably maize, preferably where the maize wild-type sequence has the sequence as shown in SEQ ID NO:3;

[0297] - D162X, preferably D162A, when the plant is from the genus Beta, preferably sugar beet, preferably where the sugar beet wild-type sequence has the sequence as shown in SEQ ID NO:16;

[0298] - D143X, preferably D143A, when the plant is from the genus Solanum, preferably potato, preferably where the common potato wild-type sequence has the sequence as shown in SEQ ID NO:17;

[0299] - D140X, D141X or D145X, preferably D140A, D141A or D145A, when the plant is from the genus Glycine, preferably soybean, preferably where the soybean wild-type sequences have the sequences as shown in SEQ ID NO:14 / 15, 12 or 13, respectively;

[0300] -D149X, preferably D149A, when the plant is from the genus Triticum, preferably wheat, preferably wherein the wild-type sugar beet sequence has a sequence as shown in SEQ ID NO: 6, 7 or 8;

[0301] -D147X, preferably D147A, when the plant is from the genus Triticum, preferably wheat, preferably wherein the wild-type sugar beet sequence has a sequence as shown in SEQ ID NO: 9, 10 or 11;

[0302] -D149X or D144X, preferably D149A or D144A, when the plant is from the genus Sorghum, preferably sorghum, preferably wherein the wild-type sorghum sequence has a sequence as shown in SEQ ID NO: 4 or 5, respectively;

[0303] wherein X is an amino acid different from D.

[0304] In certain embodiments, the mutated CPL1 protein has a sequence as shown in any one of SEQ ID NO: 80 - 87.

[0305] In certain embodiments, the wild-type CPL1 gene has or comprises:

[0306] (i) a nucleotide sequence having a cDNA or coding sequence as shown in any one of SEQ ID NO: 19 - 34;

[0307] (ii) a nucleotide sequence encoding a polypeptide having an amino acid sequence as shown in any one of SEQ ID NO: 2 - 17;

[0308] (iii) a nucleotide sequence having at least 60% identity with any one of the sequences of SEQ ID NO: 19 - 34; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity;

[0309] (iv) a nucleotide sequence encoding a polypeptide having at least 60% identity with any one of the sequences of SEQ ID NO: 2 - 17; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity;

[0310] (v) a nucleotide sequence that hybridizes under stringent hybridization conditions to the reverse complementary sequence of the nucleotide sequence defined in (i) or (ii); and

[0311] (vi) A nucleotide sequence encoding a protein that is derived from a polypeptide encoded by the nucleotide sequence of any one of (i) to (v) by substitution, deletion, and / or addition of one or more amino acids.

[0312] In certain embodiments, the wild-type ERF922 gene has or comprises:

[0313] (i) A nucleotide sequence having any cDNA or coding sequence of SEQ ID NO: 52 - 65;

[0314] (ii) A nucleotide sequence encoding a polypeptide having any amino acid sequence of SEQ ID NO: 37 - 50;

[0315] (iii) A nucleotide sequence having at least 60% sequence identity with any one of the sequences of SEQ ID NO: 52 - 65; such as at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity;

[0316] (iv) A nucleotide sequence encoding a polypeptide having at least 60% sequence identity with any one of the sequences of SEQ ID NO: 37 - 50; such as at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity;

[0317] (v) A nucleotide sequence that hybridizes under stringent hybridization conditions to the reverse complementary sequence of the nucleotide sequence defined in (i) or (ii); and

[0318] (vi) A nucleotide sequence encoding a protein that is derived from a polypeptide encoded by the nucleotide sequence of any one of (i) to (v) by substitution, deletion, and / or addition of one or more amino acids.

[0319] In certain embodiments, the wild-type CPL1 or ERF922 protein comprises a sequence that is preferably at least 95% identical to any one of the sequences of SEQ ID NO: 2 - 17 or 37 - 50 over its full length, or a sequence encoded by a sequence that is preferably at least 95% identical to any one of the sequences of SEQ ID NO: 19 - 34 or 52 - 65 over its full length.

[0320] On the one hand, the present invention provides a nucleic acid which, after transcription or expression in a plant or after silencing in a plant, is suitable for enhancing pathogen resistance or tolerance caused, for example, by mutant CPL1 and / or ERF922 genes. On the other hand, an endogenous DNA sequence in a plant genome or in a plant haploid inducer genome, which is identical to one of the nucleic acids according to the present invention, can also be modified such that the property of mediating pathogen resistance or tolerance or increasing pathogen resistance or tolerance is achieved after transcription or expression of the endogenous DNA sequence.

[0321] The nucleic acid of the present invention is preferably an isolated nucleic acid extracted from its natural or original environment (genetic background). The nucleic acid can be double-stranded or single-stranded, linear or circular. Thus it can be of genomic DNA, synthetic DNA, cDNA or RNA type (such as lncRNA, siRNA or miRNA), where the nucleobase uracil appears instead of the nucleobase thymine in RNA.

[0322] The nucleic acid according to the present invention can be used as a transgene. On the other hand, an endogenous DNA sequence in a plant genome that is identical to one of the nucleic acids according to the present invention can also be modified, whereby pathogen resistance or tolerance, for example caused by mutant CPL1 and / or ERF922 genes, can be enhanced after transcription or expression of the endogenous DNA sequence or after silencing of the endogenous DNA sequence.

[0323] On the one hand, the present invention relates to an (isolated) polynucleic acid comprising or consisting of any one of the sequences of SEQ ID NO: 19-35 or 52-79, its complementary sequence or reverse complementary sequence.

[0324] On the one hand, the present invention relates to a polynucleic acid encoding a polypeptide having any one of the sequences of SEQ ID NO: 2-17, 37-50 or 80-87, its complementary sequence or reverse complementary sequence.

[0325] On the one hand, the present invention relates to a polynucleic acid comprising:

[0326] (i) a nucleotide sequence having any one of the sequences of SEQ ID NO: 19-35 or 52-79, its complementary sequence or reverse complementary sequence;

[0327] (ii) a nucleotide sequence encoding a polypeptide having any one of the amino acid sequences of SEQ ID NO: 2-17, 37-50 or 80-87; its complementary sequence or reverse complementary sequence;

[0328] (iii) a nucleotide sequence having at least 60% identity (preferably over the entire length) to any of the sequences of SEQ ID NOs: 19-35 or 52-79; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity; its complementary sequence or reverse complementary sequence;

[0329] (iv) a nucleotide sequence encoding a polypeptide having at least 60% identity (preferably over the entire length) to any of the sequences of SEQ ID NOs: 2-17, 37-50 or 80-87; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity; its complementary sequence or reverse complementary sequence;

[0330] (v) a nucleotide sequence that hybridizes under stringent hybridization conditions to the reverse complementary sequence of the nucleotide sequence defined in (i) or (ii), its complementary sequence or reverse complementary sequence;

[0331] (vi) a nucleotide sequence encoding a protein that is derived from the polypeptide encoded by the nucleotide sequence of any one of (i) to (v) by substitution, deletion and / or addition of one or more amino acids, its complementary sequence or reverse complementary sequence; and

[0332] (vii) a functional fragment of any one of (i) to (vi) above.

[0333] On the one hand, the present invention relates to a polynucleotide comprising:

[0334] (i) a nucleotide sequence having any of the sequences of SEQ ID NOs: 19-34 or 52-65, its complementary sequence or reverse complementary sequence;

[0335] (ii) a nucleotide sequence encoding a polypeptide having any of the amino acid sequences of SEQ ID NOs: 2-17 or 37-50, its complementary sequence or reverse complementary sequence;

[0336] (iii) a nucleotide sequence having at least 60% identity (preferably over the entire length) to any of the sequences of SEQ ID NOs: 19-34 or 52-65; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity, its complementary sequence or reverse complementary sequence, preferably wherein the nucleotide sequence encodes a functional polypeptide;

[0337] (iv) a nucleotide sequence encoding a polypeptide having at least 60% identity (preferably over the entire length) to any of the sequences of SEQ ID NO: 2-17 or 37-50; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity, its complementary sequence or reverse complementary sequence, preferably wherein said nucleotide sequence encodes a functional polypeptide;

[0338] (v) a nucleotide sequence that hybridizes under stringent hybridization conditions to the reverse complementary sequence of the nucleotide sequence defined in (i) or (ii), its complementary sequence or reverse complementary sequence, preferably wherein said nucleotide sequence encodes a functional polypeptide;

[0339] (vi) a nucleotide sequence encoding a protein that is derived from a polypeptide encoded by the nucleotide sequence of any one of (i) to (v) by substitution, deletion and / or addition of one or more amino acids, its complementary sequence or its reverse complementary sequence, preferably wherein said nucleotide sequence encodes a functional polypeptide; and

[0340] (vii) a functional fragment of any one of (i) to (vi) above.

[0341] On the one hand, the present invention relates to a polynucleotide comprising:

[0342] (i) a nucleotide sequence encoding a polypeptide having any of the amino acid sequences of SEQ ID NO: 80-87, its complementary sequence or reverse complementary sequence;

[0343] (ii) a nucleotide sequence encoding a polypeptide having at least 60% identity (preferably over the entire length) to any of the sequences of SEQ ID NO: 80-87; for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity or at least 95% sequence identity, its complementary sequence or reverse complementary sequence, preferably wherein said nucleotide sequence encodes a dominant negative polypeptide;

[0344] (iii) a nucleotide sequence that hybridizes under stringent hybridization conditions to the reverse complementary sequence of the nucleotide sequence defined in (i) or (ii), its complementary sequence or reverse complementary sequence, preferably wherein said nucleotide sequence encodes a dominant negative polypeptide;

[0345] (iv) a nucleotide sequence encoding a protein that is derived from a polypeptide encoded by the nucleotide sequence of any one of (i) to (iii) by substitution, deletion, and / or addition of one or more amino acids, its complementary sequence or reverse complementary sequence, preferably wherein the nucleotide sequence encodes a dominant-negative polypeptide; and

[0346] (v) a functional fragment of any one of the above (i) to (iv).

[0347] In one aspect, the present invention relates to an (isolated) polynucleotide comprising a mutated CPL1 and / or ERF922 as described elsewhere herein, for example encoding a dominant-negative CPL1 and / or ERF922, or its complementary sequence or reverse complementary sequence.

[0348] In certain embodiments, the polynucleotide is DNA. In certain embodiments, the polynucleotide is RNA. In certain embodiments, the polynucleotide is single-stranded. In certain embodiments, the polynucleotide is double-stranded. In certain embodiments, the polynucleotide is single-stranded DNA. In certain embodiments, the polynucleotide is single-stranded RNA. In certain embodiments, the polynucleotide is double-stranded DNA. In certain embodiments, the polynucleotide is double-stranded RNA.

[0349] In one aspect, the present invention relates to a polynucleotide that specifically hybridizes to a polynucleotide as described above, or its complementary sequence or reverse complementary sequence. In certain embodiments, such a polynucleotide is at least 80% identical (i.e., complementary) (preferably over the entire length), preferably at least 90% identical, such as at least 95%, 96%, 97%, 98%, 99% or 100% identical. In certain embodiments, such a polynucleotide is 100% identical (i.e., complementary).

[0350] In one aspect, the present invention relates to a first polynucleotide comprising a second polynucleotide that specifically hybridizes to a polynucleotide as described above (such as gRNA, shRNA, siRNA), or its complementary sequence or reverse complementary sequence. In certain embodiments, the second polynucleotide and the above polynucleotide are at least 80% identical (i.e., complementary) (preferably over the entire length), preferably at least 90% identical, such as at least 95%, 96%, 97%, 98%, 99% or 100% identical. In certain embodiments, such a polynucleotide is 100% identical (i.e., complementary).

[0351] In certain embodiments, the present invention relates to an RNAi molecule comprising a polynucleotide having a sequence as shown in any one of SEQ ID NO: 35, 69 or 70, its complementary sequence or reverse complementary sequence.

[0352] In certain embodiments, the present invention relates to gRNAs that comprise a sequence as shown in any one of SEQ ID NO:67, 68, or 71-79.

[0353] In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 50,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 40,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 30,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 25,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 20,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 15,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 10,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise fewer than 5,000 nucleotides. In certain embodiments, the nucleotide molecules as described herein comprise at least 100 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 50,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 40,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 30,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 25,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 20,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 15,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 10,000 nucleotides. In certain embodiments, the nucleic acid molecules as described herein comprise at least 100 nucleotides and fewer than 5,000 nucleotides.

[0354] In certain embodiments, the polynucleic acid comprises at least 15 nucleotides, such as 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides, such as at least 30, 35, 40, 45, or 50 nucleotides, such as at least 100, 200, 300, or 500 nucleotides.

[0355] In certain embodiments, the polynucleic acids as described herein are primers or probes. In certain embodiments, the polynucleic acid is (and includes) a primer of 10 to 80 nucleotides, such as (and includes) 15 to 50 nucleotides or 15 to 25 nucleotides. In certain embodiments, the polynucleic acid is (and includes) a probe of 10 to 500 nucleotides, such as (and includes) 50 to 400 nucleotides or 100 to 250 nucleotides.

[0356] In certain embodiments, the primer or probe is capable of specifically detecting the polynucleic acids of the present invention, such as specifically hybridizing with the polynucleic acids of the present invention. In certain embodiments, the primer or probe is capable of hybridizing with any unique nucleotide fragment or portion or its complementary sequence or reverse complementary sequence of SEQ ID NOs: 19 - 34 or 52 - 68. In certain embodiments, the primer or probe is capable of hybridizing with a unique nucleotide fragment or portion or complementary sequence or reverse complementary sequence of a polynucleic acid encoding any one of the proteins of SEQ ID NOs: 2 - 17, 37 - 50 or 80 - 87. In certain embodiments, the primer or probe is capable of specifically hybridizing or capable of specifically detecting a polynucleic acid encoding a dominant negative CPL1 or ERF922 protein (i.e., capable of distinguishing between wild - type and dominant negative protein - coding sequences).

[0357] It should be understood that in embodiments involving a set of forward and reverse primers, only one of the two primers (forward or reverse) may need to be capable of distinguishing between wild - type and mutant, and thus can be unique. The other primer may or may not be capable of distinguishing between wild - type and mutant, and thus can be unique.

[0358] In certain embodiments, the polynucleic acids as described herein are RNAi polynucleic acids, siRNA or shRNA.

[0359] In certain embodiments, the polynucleic acids as described herein are guide RNAs (gRNAs).

[0360] In one aspect, the present invention relates to a vector comprising the polynucleic acids of the present invention.

[0361] As used herein, "vector" has its ordinary meaning in the art and can be, for example, a plasmid, cosmid, phage, or expression vector, transformation vector, shuttle vector, or cloning vector; it can be double-stranded or single-stranded, linear or circular; or it can transform a prokaryotic or eukaryotic host by integration into its genome or episomally. The nucleic acids according to the invention are preferably operably linked in a vector to one or more regulatory sequences which allow transcription and optionally expression in a prokaryotic or eukaryotic host cell. The regulatory sequences, preferably DNA, can be homologous or heterologous to the nucleic acids according to the invention. For example, the nucleic acids are under the control of a suitable promoter or terminator. Suitable promoters can be constitutively inducible promoters (e.g., the 35S promoter from "Cauliflower mosaic virus" (Odell et al., 1985)); those tissue-specific promoters are particularly suitable (e.g., pollen-specific promoters, Chen et al. (2010), Zhao et al. (2006) or Twell et al. (1991)), or developmentally specific (e.g., flowering-specific promoters). Suitable promoters can also be synthetic or chimeric promoters which do not exist in nature, are composed of multiple elements, and contain a minimal promoter, as well as at least one cis-regulatory element upstream of the minimal promoter, serving as a binding site for specific transcription factors. Chimeric promoters can be designed according to the desired properties and induced or repressed by different factors. Examples of such promoters can be found in Gurr & Rushton (2005) or Venter (2007). For example, a suitable terminator is the nos-terminator (Depicker et al., 1982). The vector can be introduced by conjugation, mobilization, gene gun transformation, Agrobacterium-mediated transformation, transfection, transduction, vacuum infiltration, or electroporation.

[0362] As used herein, the term "operably linked" or "operably connected" means being linked in a common nucleic acid molecule in such a way that the linked elements are positioned and oriented relative to each other such that transcription of the nucleic acid molecule can occur. DNA operably linked to a promoter is under the transcriptional control of that promoter.

[0363] In certain embodiments, the vector is a conditional expression vector. In certain embodiments, the vector is a constitutive expression vector. In certain embodiments, the vector is a tissue-specific expression vector, such as a pollen-specific expression vector. In certain embodiments, the vector is an inducible expression vector. All of these vectors are well known in the art.

[0364] The method for preparing the vector is a common method for those skilled in the art (Sambrook et al., 2001).

[0365] Also contemplated herein are host cells, such as plant cells, that contain a nucleic acid or a vector as described herein. The host cell can contain the nucleic acid as an extrachromosomal (episomal) replicating molecule, or contain the nucleic acid integrated into the nuclear or plastid genome of the host cell, or as an introduced chromosome, such as a minichromosome.

[0366] The host cell can be a prokaryotic cell (e.g., a bacterium) or a eukaryotic cell (e.g., a plant cell or a yeast cell). For example, the host cell can be an Agrobacterium, such as Agrobacterium tumefaciens or Agrobacterium rhizogenes. Preferably, the host cell is a plant cell.

[0367] The nucleic acid or the vector as described herein can be introduced into the host cell by well-known methods, which can depend on the selected host cell, including, for example, conjugation, mobilization, gene gun transformation, Agrobacterium-mediated transformation, transfection, transduction, vacuum infiltration, or electroporation. In particular, the methods for introducing the nucleic acid or the vector into Agrobacterium cells are well-known to those skilled in the art and can include conjugation or electroporation methods. The methods for introducing the nucleic acid or the vector into plant cells are also known (Sambrook et al., 2001) and can include various transformation methods, such as gene gun transformation and Agrobacterium-mediated transformation.

[0368] In one aspect, the invention relates to a kit that contains a polynucleotide, such as a primer (including forward and / or reverse primers) and / or a probe, a vector, or a host cell of the invention as described herein. The kit can also include instructions for use.

[0369] In one aspect, the invention relates to a method for identifying a plant, a plant part, or a plant population that has increased resistance and / or tolerance to a pathogen, including screening and / or identifying in the plant, the plant part, or the plant population a CPL1 and / or ERF922 protein and / or gene that has reduced or eliminated expression, activity, and / or stability, or screening and / or identifying a mutation as elsewhere herein.

[0370] On the one hand, the present invention relates to a method for identifying plants, plant parts or plant populations having increased resistance and / or tolerance to pathogens, comprising isolating genetic material (e.g., genomic DNA or mRNA) from the plants, plant parts or plant populations, and screening and / or identifying in the plants, plant parts or plant populations CPL1 and / or ERF922 proteins and / or genes having reduced or eliminated expression, activity and / or stability, or screening and / or identifying mutations as described elsewhere herein.

[0371] On the one hand, the present invention relates to a method for selecting plants, plant parts or plant populations having increased resistance and / or tolerance to pathogens, comprising screening and / or identifying in the plants, plant parts or plant populations CPL1 and / or ERF922 proteins and / or genes having reduced or eliminated expression, activity and / or stability, or screening and / or identifying mutations as described elsewhere herein; and selecting plants, plant parts or plant populations having reduced or eliminated expression, activity and / or stability of CPL1 and / or ERF922 proteins or having mutations as described elsewhere herein.

[0372] On the one hand, the present invention relates to a method for selecting plants, plant parts or plant populations having increased resistance and / or tolerance to pathogens, comprising isolating genetic material (e.g., genomic DNA or mRNA) from the plants, plant parts or plant populations, and screening and / or identifying in the plants, plant parts or plant populations CPL1 and / or ERF922 proteins and / or genes having reduced or eliminated expression, activity and / or stability, or screening and / or identifying mutations as described elsewhere herein; and selecting plants, plant parts or plant populations having reduced or eliminated expression, activity and / or stability of CPL1 and / or ERF922 proteins or having mutations as described elsewhere herein.

[0373] Methods for screening for the presence of CPL1 and / or ERF922 genes having reduced expression, activity or stability or CPL1 and / or ERF922 genes having mutations as described herein are known in the art. Non-limitingly, screening can cover or include sequencing, hybridization-based methods (e.g., (dynamic) allele-specific hybridization, molecular beacons, SNP microarrays), enzyme-based methods (e.g., PCR, KASP (competitive allele-specific PCR), RFLP, ALFP, RAPD, Flap endonuclease, primer extension, 5'-nuclease, oligonucleotide ligation assay), post-amplification methods based on DNA physical properties (such as single-strand conformation polymorphism, temperature gradient gel electrophoresis, denaturing high performance liquid chromatography, high-resolution melting of intact amplicons, use of DNA mismatch binding proteins, SNPlex, surveyor nuclease assay), etc.

[0374] Aspects and embodiments of the present invention are further supported by the following non-limiting examples. The following examples, including the experiments conducted and the results obtained, are for illustrative purposes only and do not constitute a limitation on the present invention. Example

[0375] Example 1: Identification of CPL1 Genes

[0376] Using a homology search with the Arabidopsis AtCPL1 protein sequence (SEQ ID NO: 1), CPL1-encoding genes (SEQ ID NOs: 2 - 17) have been identified in a variety of crop plants.

[0377] The selected CPL1 proteins (SEQ ID NOs: 2 - 17) have 46% to 60% identity with the Arabidopsis AtCPL1 protein at the amino acid level (see Figure 1 ; Table 1). The coding sequences (CDS) (SEQ ID NOs: 19 - 34) of the selected CPL1 genes have 53% to 66% identity with the Arabidopsis AtCPL1 coding sequence (SEQ ID NO: 18) at the nucleotide level (also see Figure 2 ; Table 2). Soybean (Glycine max) contains four paralogous CPL1 sequences, and there are two CPL1 paralogs each in maize (Zea mays) and sorghum (Sorghum bicolor). There is one CPL1 gene each on chromosomes 2A, 2B, 2D, 6A, 6B, and 6D of wheat (Triticum aestivum).

[0378] Table 1: Percentage identity between different protein sequences of the revealed CPL1 genes and comparison with the Arabidopsis AtCPL1 protein sequence

[0379]

[0380]

[0381] Table 2: Percentage identity between different coding sequences of the revealed CPL1 genes and comparison with the Arabidopsis AtCPL1 coding sequence

[0382]

[0383] Example 2: Fungal Resistance in Maize (Zea mays) with RNAi or Mutated CPL1 Genes

[0384] RNAi-based silencing constructs targeting the maize ZmCPL1.1 and ZmCPL1.2 genes ( Figure 1-2)has been developed and stably transformed into maize plants of the A188 genotype.

[0385] The ZmCPL1 silencing sequence (SEQ ID NO:35) was specifically selected to have a very high homology with the selected ZmCPL1 gene (100% identity with ZmCPL1.1, >95% identity with ZmCPL1.2), while avoiding large stretches of homology (<20bp) with other sequences in the maize genome. The vector construct used for transformation is as Figure 3 shown. Transgenic maize plants of the T1 generation for detecting gene segregation, homozygous T2 lines, and their respective unpaired sister lines (null segregants) were tested for resistance to Setosphaeria turcica (northern corn leaf blight) (NCLB) in the greenhouse and to Fusarium in the field.

[0386] These experiments showed that downregulation of the ZmCPL1 gene led to increased resistance to hemibiotrophic fungi, and downregulation of ZmCPL1 did not cause any growth retardation.

[0387] There are multiple ways to downregulate gene expression. These include, for example, the expression of RNAi constructs or microRNA constructs or the modification of the promoter or other regulatory elements of the gene. For the downregulation of the CPL1 gene, in addition to the above methods, a dominant negative allele of CPL1 was also expressed. It has been shown that a point mutation D128A in the conserved DXDXT motif of the catalytic phosphatase domain results in a dominant negative form of the Arabidopsis protein AtCPL4, which is involved in normal plant growth and plant development (Fukudome et al.;2014). Overexpression of the dominant negative allele AtCPL4_D128A in Arabidopsis is lethal and phenotypically similar to the homozygous lethal AtCPL4 knockout line. Arabidopsis plants with the AtCPL4_RNAi construct are viable with a mildly toxic phenotype. This indicates that strong overexpression of the dominant negative AtCPL4 allele is similar to a complete knockout of AtCPL4.

[0388] To achieve downregulation of the CPL1 gene, a dominant-negative allele under the control of a strong overexpression promoter or under the control of a weaker promoter such as the native promoter was used to obtain a dominant-negative allele of the selected CPL1 gene. In one embodiment, a DXDXT motif mutation similar to D128A in AtCPL4 was used. Examples of DXDXT motif modifications are D148A in ZmCPL1.1 (SEQ ID NO:80) and D149A in ZmCPL1.2 (SEQ ID NO:81). In other important crops, these positions correspond to D162A in the g16374.t1_protein of sugar beet (CPL1 homolog in sugar beet; SEQ ID NO:82), D143A in the PGSC0003DMT400057635_protein of potato (CPL1 homolog in potato; SEQ ID NO:83), D141A in the Glyma.07g194800.1_protein of soybean (CPL1 homolog in soybean; SEQ ID NO:84), D149A in the TraesCS2A02G373400.3_protein of wheat (CPL1 homolog in wheat; SEQ ID NO:85), D147A in the TraesCS6B02G264000.4_protein of wheat (CPL1 homolog in wheat; SEQ ID NO:86), and D149A in the Sobic.004G227600.1_protein of sorghum (CPL1 homolog in sorghum; SEQ ID NO:87).

[0389] Thus, downregulation of the CPL1 gene can also be achieved by introducing a point mutation in the native CPL1 gene that results in a dominant-negative allele and maintaining the mutation in the heterozygous state. The resistance effect of the dominant-negative allele of CPL1 will be genetically dominant, which is beneficial for breeding resistant hybrid crops compared to, for example, the requirement for recessive mutations in the promoters of both parents of the hybrid. If paralogous CPL1 genes exist, such as in maize, wheat, soybean, and sorghum, engineering resistant plants with a dominant-negative CPL1 allele requires less effort compared to, for example, the desired effort to downregulate all CPL1 paralogs simultaneously by promoter modification.

[0390] Example 3: Fungal resistance in wheat (Triticum aestivum) by VIGS method targeting CPL1

[0391] Two silencing constructs targeting all six paralogs / homologs of CPL1 have been developed for virus-induced gene silencing (VIGS) experiments in wheat. The silencing sequences were specifically designed to have a high homology (>83% identity) with all six wheat CPL1 homologs, while avoiding large stretches of homology (<20 bp) with other coding sequences in the wheat genome. For the VIGS experiments, an appropriate vector system was used. Here, the TaCPL1 silencing sequences (SEQ ID NO: 69 and 70) were inserted individually. Virus particles carrying the silencing sequences were assembled by Agrobacterium co-infiltration transformation into Nicotiana benthamiana. Subsequently, the leaves of the wheat cultivar Passat (KWS, Einbeck, Germany) were transfected with the sap extracted from the infiltrated N. benthamiana leaves. Fourteen days after transfection with the viral variant, the wheat plants were infected with a pathogen suspension, for example with fungal spores of Zymoseptoria tritici, which is the causative agent of wheat leaf blotch. Subsequently, the plants were kept under conditions that supported pathogen infection and scored for pathogen infection.

[0392] Compared to mock-inoculated, untreated or wheat plants infected with an empty vector control, wheat plants infected with the CPL1 silencing construct showed a significant reduction in the fungal pathogen ( Figure 4 ).

[0393] This indicates that silencing of all CPL1 homologs in wheat leads to an increased resistance to insects or pathogens, including hemibiotrophic fungal pathogens.

[0394] As a complement to the fungal pathogen assays, insect resistance assays were performed. It was expected that silencing of CPL1 would also affect the susceptibility of plants, especially wheat plants, to insect infestation, i.e., modification of the CPL1 protein or reduction of CPL1 gene expression would increase the insect resistance of the plants. Therefore, the CPL1 silencing construct as well as the above-described dominant negative alleles can also be used as a method for increasing insect resistance.

[0395] Example 4: Fungal resistance in wheat (Triticum aestivum) by the KO method of the CPL1 gene

[0396] To confirm the correlation between the CPL1 gene and fungal resistance, EMS or ENU was used as a mutagen to introduce (knockout) mutations in this gene by the TILLING method. Subsequently, the selected plants were self-pollinated to generate homozygous mutants. The increased resistance of the homozygous CPL1 mutants to fungal pathogens was analyzed. If there are multiple paralogs in the plant, the cumulative effect of parallel knockout of multiple paralogous genes is expected. Complete knockout of the CPL1 gene and not just downregulation of its expression results in delayed flowering. Therefore, downregulation of CPL1 gene expression can be the preferred technical guidance.

[0397] As an alternative to the TILLING method, genome editing was used to generate knockout mutations in the CPL1 gene. Modification of the CPL1 gene regulatory elements by genome editing was used to downregulate CPL1 gene expression. Regarding the delayed flowering in CPL1 complete knockout plants, downregulation of expression prevented this side effect of the CPL1 resistance method.

[0398] For genome editing of CPL1, target validation has been performed. Maize protoplasts were co-transfected with two constructs. One construct carried the constitutively expressed MAD7 gene, which has nuclease activity if expressed, and the second construct carried a single guide RNA (crRNA) constitutively expressed for each target site. After transfection, the protoplasts were cultured for 24 hours.

[0399] The expression of a fluorescent marker gene carried on the same plasmid as the MAD7 gene was used to measure the transfection efficiency of the protoplast samples. The protoplasts were counted using a flow cytometer.

[0400] Genomic DNA was extracted from the protoplast samples, and the DNA samples were PCR amplified to obtain amplicons flanking the target sequences. Amplicon deep sequencing was used to determine the cleavage rate at each site. The INDEL frequencies are shown in Table 3. Each treatment was repeated 3 times, and the data are shown in the following table. The INDEL frequencies and standard errors are based on the raw data. The second set of values was adjusted according to the protoplast transfection efficiency.

[0401] Table 3

[0402]

[0403]

[0404] Example 5: Identification of the ERF922 Gene

[0405] Using homology searches with the rice ERF922 protein sequence, ERF922-encoding genes have been identified in a variety of crop plants, which encode the ERF922 protein sequence (SEQ ID NO: 36-50). The corresponding DNA coding regions are shown (SEQ ID NO: 51-65). In maize (Zea mays), sorghum (Sorghum bicolor), sugar beet (Beta vulgaris) and potato (Solanum tuberosum), the protein of the ERF922 gene is encoded by one gene, ERF922-I. In the gramineous wheat (Triticum aestivum) and rye (Secale graine), in addition to ERF922-I, there is also a second ERF992-like gene, which is also highly homologous to OsERF922, but belongs to a second ERF922-like protein named ERF922-II. In wheat, the ERF922-I and ERF922-II genes are located on chromosomes 3 and 2 as single alleles in the A, B and D genomes ( Figure 5 and 6 ). Unique but related ERF922 alleles are present in rye and wheat plants as Figure 5 shown.

[0406] The transcription of OsERF922 is induced by fungal infection and abiotic stresses (salt and drought) in rice leaves. Wheat mRNA seq data analysis shows that the A, B and D alleles of ERF922-I are expressed only in wheat ears after Fusarium infection. Compared with ERF922-II, the expression of ERF922-I in uninfected wheat is detected only in the rachis and florets of wheat ears and is absent in other tissues. ERF922-II transcription can be detected in seedlings, roots, shoots, flag leaves and spikelets, rachises and inflorescence florets. After fungal infection or stress treatment of wheat plants in leaves, the expression of the ERF922-I allele is not detected. However, the three alleles of ERF922-II are induced by both biotic and abiotic stresses in all tissues. ERF922-II transcription is induced by Fusarium in wheat ears and by Septoria tritici, Puccinia and powdery mildew infection in wheat leaves. In addition, the expression of ERF922-II is induced by cold. The ERF922 genes in rye and wheat are paralogs. In order to improve wheat resistance to leaf diseases, the expression of ERF922-II must be reduced. Reducing ERF922-I or ERF922-II increases wheat resistance to Fusarium head blight.

[0407] Example 6: Fungal resistance in wheat (Triticum aestivum) by VIGS method targeting ERF922-I and ERF922-II

[0408] For virus-induced gene silencing (VIGS) experiments in wheat, two silencing constructs (TaERF922-3A-I, TaERF922-3B-I, TAERF922-3D-I) targeting all six alleles of TaERF922-I on chromosome 3 and two silencing constructs (TaERF922-2A-II, TaERF922-2B-II, TAERF922-2D-II) targeting all six alleles of TaERF922-II on chromosome 2 have been developed. The silencing sequences were specifically designed to have high homology (>92% identity) with all six wheat ERF922-I homologs or all six wheat ERF922-II homologs, while avoiding large stretches of homology (<20 bp) with other coding sequences in the wheat genome. For VIGS experiments, an appropriate vector system was used.

[0409] Here, the TaERF922-I silencing sequences of fragments TaERF922-3A_fragA and TaERF922-3A_fragB and the TaERF922-II silencing sequences of TaERF922-2A_fragA and TaERF922-2A_fragB were inserted respectively. The sequences of TaERF922-3A_fragA and TaERF922-3A_fragB are identical to positions 70 - 288 and 368 - 558 of SED ID NO:58 - 60. The sequences of TaERF922-2A_fragA and TaERF922-2A_fragB are identical to positions 21 - 179 and 611 - 766 of SED ID NO:61 - 63. Virus particles carrying the silencing sequences were assembled into Nicotiana benthamiana by Agrobacterium co-infiltration transformation. Subsequently, the leaves of wheat cultivar Taifun (KWS, Einbeck, Germany) were transfected with the juice extracted from the infiltrated Nicotiana benthamiana leaves. 14 days after transfection with the virus variant, the leaves of the wheat plants were infected with fungal spores of Zymoseptoria tritici, which is the pathogen of wheat leaf blotch, and the wheat ears were infected with spores of Fusarium graminearum, which is the pathogen of Fusarium head blight. Subsequently, the plants were kept under conditions supportive of pathogen infection and scored for pathogen infection.

[0410] Compared with wheat plants inoculated mock, untreated or infected with empty vector control, wheat plants infected with ERF922-I or ERF922-II silencing constructs showed significantly reduced Septoria tritici blotch ( Figure 7 ) and Fusarium head blight ( Figure 8 ).

[0411] This indicates that silencing of all ERF922 homologs in wheat leads to increased resistance to fungal pathogens, including hemibiotrophic and necrotrophic fungal pathogens.

[0412] Example 7: Fungal resistance in maize (Zea mays) by RNAi approach targeting ERF922

[0413] After selecting the maize ZmERF922 gene for testing, improvement of resistance was confirmed by downregulating the expression of the ERF922-I gene. An RNAi-based silencing construct targeting the maize ERF922-I gene ( Figure 9 , SEQ ID NO:66) was constructed and stably transformed into the A188 genotype of maize by Agrobacterium tumefaciens-mediated transformation. Transgenic maize plants of the MTR554-T-015 line and the respective unpaired sister line MTR554-T-015 (non-transgenic) of the T1 generation with gene segregation were identified by PCR. The resistance of MTR554-T015 plants to Setosphaeria turcica, the pathogen causing northern corn leaf blight (NCLB), was tested in the greenhouse using genotype A188. The plant height was additionally measured before inoculation, and no differences were detected (Table 4).

[0414] Table 4: The ERF922 RNAi line MTR554-T-015 shows enhanced resistance to northern corn leaf blight compared to the transformed genotype A188 and unpaired MTR554-T-015 (non-transgenic)

[0415]

[0416] 1 Plant size measured before inoculation

[0417] In the resistance assay, the fifth leaf of each plant was spray-inoculated with a mixture of two isolates (race 0) of Setosphaeria turcica at 5,000 spores / ml. The transgenic ERF922RNAi plants of the MTR554-T-015 line showed enhanced resistance after visual disease scoring of the fifth leaf at 18 and 23 days post inoculation (dpi). Compared to the transformed genotype A188 and the unpaired sister line MTR554-T-015 (non-transgenic), the symptoms of ERF922 RNAi plants were reduced by 30% and 23% at 18 dpi and 23 dpi, respectively. At 24 dpi, the fifth leaf of each plant was sampled, dried, and fungal biomass was determined by qPCR from the isolated genomic DNA. The fungal biomass of the transgenic MTR554-T-015 line was reduced by 14% compared to the average of the fungal biomass in A188 and MTR554-T-015 (non-transgenic). These experiments indicate that downregulation of the ZmERF922 gene leads to increased resistance to hemibiotrophic fungi in maize, and downregulation of ZmERF922 does not cause growth retardation.

[0418] Example 8: Fungal Resistance in Wheat (Triticum aestivum) by ERF922 Genome Editing

[0419] Genome editing was applied to downregulate the expression of ERF922-I and ERF922-II in wheat. Wheat protoplasts were co-transfected with two constructs; one construct carried the constitutively expressed RR-Cpf1 gene, and the second construct carried a single guide RNA constitutively expressed for each target site. The protospacers for the target sites crGEP239 and crGEP243 are shown as SEQ ID NO:67 and 68. After transfection, the protoplasts were cultured for 24 hours. The transfection efficiency of the protoplast samples was measured using the expression of a fluorescent marker gene carried on the same plasmid as the RR-Cpf1 gene. The protoplasts were counted using a flow cytometer.

[0420] Genomic DNA was extracted from the protoplast samples, and the DNA samples were PCR amplified to obtain amplicons flanking the target sequences. The cleavage rate at each site in the A and B genomes of wheat was determined using amplicon deep sequencing. The INDEL frequencies are shown in Table 5.

[0421] Table 5: Detection of gRNAs (pGE745 - 750) in wheat for gene disruption. InDel frequencies are given as percentages %.

[0422] gene construct target target annotation INDEL frequency (%) Ta-ERF922 pGEP745 crGEP239 A, only B1 A4.0, B2.4 Ta-ERF922 pGEP746 crGEP240 A, only B1 A1.2, B3.5 Ta-ERF922 pGEP747 crGEP241 A, only B1 A0.0, B0.0 Ta-ERF922 pGEP748 crGEP242 only B2 0.0 Ta-ERF922 pGEP749 crGEP243 only B2 0.6 Ta-ERF922 pGEP750 crGEP244 only B2 0.1 Sequence Listing <110> KWS Saat SE & Co. KG <120>Enhancing Crop Disease Resistance through Downregulation of Repressor Genes <130>KWS0346PCT <150>US 62 / 916,578 <151>2019-10-17 <160>87 <170>PatentIn version 3.5 <210>1 <211>967 <212>PRT <213>Arabidopsis thaliana <400>1 Met Tyr Ser Asn Asn Arg Val Glu Val Phe His Gly Asp Gly Arg Leu 1 5 10 15 Gly Glu Leu Glu Ile Tyr Pro Ser Arg Glu Leu Asn Gln Gln Gln Asp 20 25 30 Asp Val Met Lys Gln Arg Lys Lys Lys Gln Arg Glu Val Met Glu Leu 35 40 45 Ala Lys Met Gly Ile Arg Ile Ser His Phe Ser Gln Ser Gly Glu Arg 50 55 60 Cys Pro Pro Leu Ala Ile Leu Thr Thr Ile Ser Ser Cys Gly Leu Cys 65 70 75 80 Phe Lys Leu Glu Ala Ser Pro Ser Pro Ala Gln Glu Ser Leu Ser Leu 85 90 95 Phe Tyr Ser Ser Cys Leu Arg Asp Asn Lys Thr Ala Val Met Leu Leu 100 105 110 Gly Gly Glu Glu Leu His Leu Val Ala Met Tyr Ser Glu Asn Ile Lys 115 120 125 Asn Asp Arg Pro Cys Phe Trp Ala Phe Ser Val Ala Pro Gly Ile Tyr 130 135 140 Asp Ser Cys Leu Val Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe 145 150 155 160 Asp Leu Asp Glu Thr Leu Val Val Ala Asn Thr Met Arg Ser Phe Glu 165 170 175 Asp Lys Ile Asp Gly Phe Gln Arg Arg Ile Asn Asn Glu Met Asp Pro 180 185 190 Gln Arg Leu Ala Val Ile Val Ala Glu Met Lys Arg Tyr Gln Asp Asp 195 200 205 Lys Asn Leu Leu Lys Gln Tyr Ile Glu Ser Asp Gln Val Val Glu Asn 210 215 220 Gly Glu Val Ile Lys Val Gln Ser Glu Ile Val Pro Ala Leu Ser Asp 225 230 235 240 Asn His Gln Pro Leu Val Arg Pro Leu Ile Arg Leu Gln Glu Lys Asn 245 250 255 Ile Ile Leu Thr Arg Ile Asn Pro Met Ile Arg Asp Thr Ser Val Leu 260 265 270 Val Arg Met Arg Pro Ser Trp Glu Glu Leu Arg Ser Tyr Leu Thr Ala 275 280 285 Lys Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala Glu Arg 290 295 300 Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Glu Gly Asn Leu 305 310 315 320 Ile Asn Thr Asn Asp Leu Leu Ala Arg Ile Val Cys Val Lys Ser Gly 325 330 335 Phe Lys Lys Ser Leu Phe Asn Val Phe Leu Asp Gly Thr Cys His Pro 340 345 350 Lys Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp Glu Lys 355 360 365 Asp Gln Pro Arg Val His Val Val Pro Ala Phe Ala Pro Tyr Tyr Ser 370 375 380 Pro Gln Ala Glu Ala Ala Ala Thr Pro Val Leu Cys Val Ala Arg Asn 385 390 395 400 Val Ala Cys Gly Val Arg Gly Gly Phe Phe Arg Asp Phe Asp Asp Ser 405 410 415 Leu Leu Pro Arg Ile Ala Glu Ile Ser Tyr Glu Asn Asp Ala Glu Asp 420 425 430 Ile Pro Ser Pro Pro Asp Val Ser His Tyr Leu Val Ser Glu Asp Asp 435 440 445 Thr Ser Gly Leu Asn Gly Asn Lys Asp Pro Leu Ser Phe Asp Gly Met 450 455 460 Ala Asp Thr Glu Val Glu Arg Arg Leu Lys Glu Ala Ile Ser Ala Ser 465 470 475 480 Ser Ala Val Leu Pro Ala Ala Asn Ile Asp Pro Arg Ile Ala Ala Pro 485 490 495 Val Gln Phe Pro Met Ala Ser Ala Ser Ser Val Ser Val Pro Val Pro 500 505 510 Val Gln Val Val Gln Gln Ala Ile Gln Pro Ser Ala Met Ala Phe Pro 515 520 525 Ser Ile Pro Phe Gln Gln Pro Gln Gln Pro Thr Ser Ile Ala Lys His 530 535 540 Leu Val Pro Ser Glu Pro Ser Leu Gln Ser Ser Pro Ala Arg Glu Glu 545 550 555 560 Gly Glu Val Pro Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu 565 570 575 Leu Ile Leu Gln His Gly Gln Asp Thr Arg Asp Pro Ala Pro Ser Glu 580 585 590 Pro Ser Phe Pro Gln Arg Pro Pro Val Gln Ala Pro Pro Ser His Val 595 600 605 Gln Ser Arg Asn Gly Trp Phe Pro Val Glu Glu Glu Met Asp Pro Ala 610 615 620 Gln Ile Arg Arg Ala Val Ser Lys Glu Tyr Pro Leu Asp Ser Glu Met 625 630 635 640 Ile His Met Glu Lys His Arg Pro Arg His Pro Ser Phe Phe Ser Lys 645 650 655 Ile Asp Asn Ser Thr Gln Ser Asp Arg Met Leu His Glu Asn Arg Arg 660 665 670 Pro Pro Lys Glu Ser Leu Arg Arg Asp Glu Gln Leu Arg Ser Asn Asn 675 680 685 Asn Leu Pro Asp Ser His Pro Phe Tyr Gly Glu Asp Ala Ser Trp Asn 690 695 700 Gln Ser Ser Ser Arg Asn Ser Asp Leu Asp Phe Leu Pro Glu Arg Ser 705 710 715 720 Val Ser Ala Thr Glu Thr Ser Ala Asp Val Leu His Gly Ile Ala Ile 725 730 735 Lys Cys Gly Ala Lys Val Glu Tyr Lys Pro Ser Leu Val Ser Ser Thr 740 745 750 Asp Leu Arg Phe Ser Val Glu Ala Trp Leu Ser Asn Gln Lys Ile Gly 755 760 765 Glu Gly Ile Gly Lys Ser Arg Arg Glu Ala Leu His Lys Ala Ala Glu 770 775 780 Ala Ser Ile Gln Asn Leu Ala Asp Gly Tyr Met Arg Ala Asn Gly Asp 785 790 795 800 Pro Gly Pro Ser His Arg Asp Ala Thr Pro Phe Thr Asn Glu Asn Ile 805 810 815 Ser Met Gly Asn Ala Asn Ala Leu Asn Asn Gln Pro Phe Ala Arg Asp 820 825 830 Glu Thr Ala Leu Pro Val Ser Ser Arg Pro Thr Asp Pro Arg Leu Glu 835 840 845 Gly Ser Met Arg His Thr Gly Ser Ile Thr Ala Leu Arg Glu Leu Cys 850 855 860 Ala Ser Glu Gly Leu Glu Met Ala Phe Gln Ser Gln Arg Gln Leu Pro 865 870 875 880 Ser Asp Met Val His Arg Asp Glu Leu His Ala Gln Val Glu Ile Asp 885 890 895 Gly Arg Val Val Gly Glu Gly Val Gly Ser Thr Trp Asp Glu Ala Arg 900 905 910 Met Gln Ala Ala Glu Arg Ala Leu Ser Ser Val Arg Ser Met Leu Gly 915 920 925 Gln Pro Leu His Lys Arg Gln Gly Ser Pro Arg Ser Phe Gly Gly Met 930 935 940 Ser Asn Lys Arg Leu Lys Pro Asp Phe Gln Arg Ser Leu Gln Arg Met 945 950 955 960 Pro Ser Ser Gly Arg Tyr Ser 965 <210> 2 <211> 932 <212> PRT <213> Zea mays <400> 2 Met Phe Lys Ser Met Val Tyr Tyr Val Asn Thr Ser Ile Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Glu Ala Ser Ala Gly Leu Thr Met Ala Ala 20 25 30 Trp Ala Arg Glu Ile Arg Val Glu Arg Ile Ser Pro Pro Ser Glu Arg 35 40 45 Cys Pro Pro Leu Ala Val Met His Thr Val Ala Val Gly Ala Arg Cys 50 55 60 Leu Val Met Glu Ser Arg Pro Pro Val Val Ala Asp Val Val Pro Pro 65 70 75 80 Leu Val Val Met His Thr Ala Cys Leu Arg Glu Asn Lys Thr Ala Val 85 90 95 Val Pro Leu Gly Asp Glu Glu Leu His Leu Val Ala Met Thr Ser Arg 100 105 110 Arg Asn Leu Thr Asn His Ala Cys Phe Trp Gly Tyr Lys Leu Pro Phe 115 120 125 Gly Leu Tyr Asn Ser Cys Leu Thr Met Leu Asn Leu Arg Cys Leu Gly 130 135 140 Ile Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Ser Arg 145 150 155 160 Ser Phe Glu Asp Arg Ile Asp Ala Leu Gln Arg Lys Leu Ser Asn Glu 165 170 175 Thr Asp Pro Gln Arg Arg Asn Gly Met Leu Ser Glu Ile Lys Arg Tyr 180 185 190 Gln Asp Asp Lys Ser Ile Leu Lys Gln Tyr Ile Glu Gly Asp Gln Val 195 200 205 Tyr Asp Asp Gly Lys Val Tyr Lys Ala Gln Pro Glu Ile Val Pro Pro 210 215 220 Leu Ser Asp Asn Gln Gln Pro Met Thr Arg Pro Val Ile Arg Leu Gln 225 230 235 240 Asp Lys Asn Ile Ile Leu Thr Arg Ile Asn Pro Leu Ile Arg Asp Thr 245 250 255 Ser Val Leu Val Cys Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr 260 265 270 Leu Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met 275 280 285 Ala Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp 290 295 300 Ser Arg Leu Ile Asn Ser Val Gln Leu His Asp Arg Met Val Cys Val 305 310 315 320 Lys Ser Gly Leu Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly Ser 325 330 335 Cys His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp 340 345 350 Asp Glu Lys Asp Gln Leu Arg Val His Val Val Pro Ala Phe Thr Pro 355 360 365 Tyr Tyr Ala Pro Gln Ala Glu Ala Asn Cys Ser Ile Pro Val Leu Cys 370 375 380 Val Ala Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Asp 385 390 395 400 Phe Asp Glu Gly Leu Leu Pro Arg Ile Ser Asn Val His Tyr Glu Asp 405 410 415 Glu Val Asn Glu Ile Ser Ala Pro Asp Val Gly Asn Tyr Leu Ile Thr 420 425 430 Asp Asp Glu Asn Val Ala Leu Val Asn Gly Asn Arg Asp Ser Leu Pro 435 440 445 Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys Glu Ala 450 455 460 Asn Ala Gln Ser Phe His Gln Thr Ala Gly Asp Phe Val Met Pro Val 465 470 475 480 Ala Pro Ala Gln Asn Phe Val Ser Thr Ser Val Ala Ser Leu Ala Pro 485 490 495 Pro Leu Gly Met Met Pro Ser Pro Phe Ser Gln Pro Val Ala Pro Pro 500 505 510 Gly Phe Ser Asp Ser Leu Gln Gly Ser Pro Ala Arg Glu Glu Gly Glu 515 520 525 Val Pro Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile 530 535 540 Leu Gln His Gly Gln Asp Thr Arg Asp Pro Thr Ser Pro Leu Pro Ala 545 550 555 560 Ile Pro Pro Val Gln Val Pro Val Pro Pro Val Gln Pro His Gly Asn 565 570 575 Trp Phe Pro Thr Glu Asp Gly Ile Asn Gln Ser Asn Leu Asn Arg Gly 580 585 590 Ser Ala Gly Phe Thr Val Glu Ser Asp Ser Ile Val Tyr Glu Lys Lys 595 600 605 Gln Pro Pro His Pro Ser Phe Phe His Gly Gly Asp Ser Pro Met Pro 610 615 620 Ser Asp Arg Phe Gly Tyr Gln Asn Gln Arg Phe Pro Ser Gln Leu Pro 625 630 635 640 His Glu Asp His Pro Met Met Gln Asn His Ala Pro Pro Lys Tyr Arg 645 650 655 Ser Phe Ser Gly Glu Glu Leu Ala Ser Trp His Val Pro Ser Ser Gln 660 665 670 Arg Asn Asn Gln Ile Glu Ser Gly Arg His Phe Ala Gln Tyr Ala Gly 675 680 685 Thr Ser Ala Gly Ile Leu Glu Gly Ile Ala Leu Lys Cys Gly Ser Lys 690 695 700 Val Glu Tyr Lys Ser Ala Leu Cys Asp Thr Ala Glu Leu Gln Phe Ser 705 710 715 720 Ile Glu Val Trp Ile Val Gly Glu Lys Val Gly Glu Gly Ile Gly Arg 725 730 735 Thr Arg Arg Glu Ala Gln Arg Gln Ala Ala Glu Met Ser Leu Arg Asn 740 745 750 Leu Ala Asn Lys Tyr Leu Ser Ser Asp Pro Asn Lys Leu Ser Asp Met 755 760 765 Lys Glu Asn Asp Phe Ser Ser Asn Arg Asn Val Phe Gly Tyr Ser Gly 770 775 780 Asn Thr Arg Asp Asp Met Leu Pro Leu Ser Ser Thr Ser Glu Glu Ser 785 790 795 800 Arg Phe Met Lys Met Glu Asn Asn Asn Ser Arg Lys Thr Gly Ser Ser 805 810 815 Val Ala Ala Leu Lys Glu Leu Cys Thr Val Glu Gly Tyr Asn Leu Val 820 825 830 Phe Gln Ala Cys Pro Ser Ser Ala Asp Gly Leu Val Gly Lys Glu Ser 835 840 845 Tyr Ala Gln Val Gln Val Gly Gly Gln Ile Leu Gly Lys Gly Val Gly 850 855 860 Leu Thr Trp Glu Glu Ala Lys Leu Gln Ala Ala Ala Glu Ala Leu Gly 865 870 875 880 Thr Leu Arg Ser Met Leu Gly Gln Leu Gly His Lys Arg Ser Gly Ser 885 890 895 Pro Arg Ser Leu Ala Pro Asn Phe Asn Lys Arg Phe Lys Pro Asp Phe 900 905 910 Pro Arg Thr Val Gln Arg Val Pro Tyr Gly Thr Tyr Ser Arg Ile Glu 915 920 925 Gly His Val Pro 930 <210> 3 <211> 934 <212> PRT <213> Zea mays <400> 3 Met Phe Lys Ser Met Val Tyr Phe Val Asn Ile Ser Ile Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Gln Ala Ser Ala Gly Leu Ala Val Ala Ala 20 25 30 Trp Ala Arg Asp Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg 35 40 45 Cys Pro Pro Leu Ala Val Met His Thr Val Ala Val Ala Ala Arg Cys 50 55 60 Leu Val Met Glu Ser Arg Pro Pro Val Ala Ala Asp Val Ala Ser Leu 65 70 75 80 Pro Leu Val Asp Met His Ala Ala Cys Leu Arg Asp Asn Lys Thr Ala 85 90 95 Val Val Ala Leu Gly Asp Glu Glu Leu His Leu Val Ala Met Thr Ser 100 105 110 Arg Arg Asn Leu Thr Asn His Ala Cys Phe Trp Gly Tyr Lys Leu Pro 115 120 125 Phe Gly Leu Tyr Asn Ser Cys Leu Thr Met Leu Asn Leu Arg Cys Leu 130 135 140 Gly Ile Val Phe Asp Leu Asp Glu Thr Leu Val Val Ala Asn Thr Ser 145 150 155 160 Arg Ser Phe Glu Asp Arg Ile Asp Ala Leu Gln Arg Lys Leu Ser Asn 165 170 175 Glu Thr Asp Pro Gln Arg Arg Asn Gly Met Leu Ser Glu Ile Lys Arg 180 185 190 Tyr Gln Asp Asp Lys Ser Ile Leu Lys Gln Tyr Ile Glu Gly Asp Gln 195 200 205 Val Tyr Asp Asp Gly Lys Val Tyr Lys Ala Gln Pro Glu Ile Val Pro 210 215 220 Pro Leu Ser Asp Asn Gln Gln Thr Met Thr Arg Pro Val Ile Arg Leu 225 230 235 240 Gln Glu Lys Asp Ile Ile Leu Thr Arg Ile Asn Pro Leu Ile Arg Asp 245 250 255 Thr Ser Val Leu Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser 260 265 270 Tyr Leu Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr 275 280 285 Met Ala Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro 290 295 300 Asp Ser Arg Leu Ile Asn Ser Val Arg Leu His Asp Arg Met Val Cys 305 310 315 320 Val Lys Ser Gly Leu Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly 325 330 335 Ser Cys His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val 340 345 350 Trp Asp Glu Lys Asp Gln Leu Arg Val His Val Val Pro Ala Phe Thr 355 360 365 Pro Tyr Tyr Ala Pro Gln Ala Glu Ala Asn Cys Ser Ile Pro Val Leu 370 375 380 Cys Val Ala Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys 385 390 395 400 Asp Phe Asp Glu Gly Leu Leu Pro Arg Ile Ser Asn Val His Tyr Glu 405 410 415 Asp Glu Val Asn Asp Ile Ser Ser Ala Pro Asp Val Gly Asn Tyr Leu 420 425 430 Ile Thr Glu Asp Glu Asn Val Ala Leu Val Asn Gly Asn Arg Asp Ser 435 440 445 Leu Pro Phe Asp Gly Met Ser Asp Ala Glu Ile Glu Arg Arg Met Lys 450 455 460 Glu Ala Asn Ala Gln Ala Phe His Gln Thr Ala Thr Asn Phe Val Met 465 470 475 480 Pro Val Ala Pro Ala Gln Asn Phe Val Ser Ser Ser Val Ala Pro Leu 485 490 495 Ala Pro Pro Leu Ser Met Met Pro Pro Pro Phe Ser Gln Pro Val Val 500 505 510 Gln Pro Gly Phe Ser Asp Pro Leu Gln Gly Ser Pro Ala Arg Glu Glu 515 520 525 Gly Glu Val Pro Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu 530 535 540 Leu Ile Leu Gln His Gly Gln Asp Thr Arg Asp Pro Thr Pro Pro His 545 550 555 560 Pro Ala Ile Pro Pro Val Gln Val Pro Val Pro Pro Val Gln Pro His 565 570 575 Gly Asn Trp Phe Pro Thr Glu Asp Gly Ile Asn Pro Ser Asn Leu Ser 580 585 590 Arg Gly Ser Ala Gly Phe Thr Val Glu Ser Asp Ser Met Pro Tyr Glu 595 600 605 Lys Lys Gln Pro Pro His Pro Ser Phe Phe His Gly Gly Asp Ser Pro 610 615 620 Met Ser Ser Asp Arg Phe Gly Tyr Gln Asn Gln Arg Phe Pro Ser Gln 625 630 635 640 Leu Pro His Thr Glu Asp His His Met Leu Gln Asn His Ala Pro Pro 645 650 655 Lys Tyr Arg Ser Phe Ser Gly Glu Glu Leu Ala Thr Arg His Val Pro 660 665 670 Ser Ser Gln Arg Asn His Ile Glu Ser Gly Arg His Phe Ser Gln Tyr 675 680 685 Ala Gly Thr Ser Ala Gly Val Leu Glu Gly Ile Ala Val Lys Cys Gly 690 695 700 Ser Lys Val Glu Tyr Arg Ser Thr Leu Cys Asp Thr Ala Glu Leu Gln 705 710 715 720 Phe Ser Ile Glu Val Trp Ile Val Gly Glu Lys Phe Gly Glu Gly Ile 725 730 735 Gly Arg Thr Arg Arg Glu Ala Gln Arg Gln Ala Ala Glu Met Ser Leu 740 745 750 Arg Asn Leu Ala Asn Lys Tyr Leu Ser Ser Asp Pro Asn Lys Leu Thr 755 760 765 Asp Met Lys Gln Asp Ala Phe Gly Ser Asn Arg Asn Ile Phe Gly Tyr 770 775 780 Ser Gly Asn Thr Arg Asp Asp Met Leu Pro Leu Ser Ser Thr Ser Glu 785 790 795 800 Glu Ser Arg Phe Met Lys Met Glu Glu Asn Asn Ser Arg Lys Thr Gly 805 810 815 Asp Ser Val Thr Ala Leu Lys Glu Leu Cys Thr Val Glu Gly Tyr Asn 820 825 830 Leu Val Phe Gln Ala Cys Pro Ser Pro Ala Asp Gly Leu Val Gly Lys 835 840 845 Glu Ser Tyr Ala Gln Val Gln Ile Gly Arg Gln Ile Leu Gly Lys Gly 850 855 860 Val Gly Leu Thr Trp Glu Glu Ala Lys Leu Gln Ala Ala Asp Glu Ala 865 870 875 880 Leu Gly Thr Leu Arg Ser Met Leu Gly Gln Leu Gly His Arg Arg Ser 885 890 895 Gly Ser Pro Arg Ser Leu Ala Pro Asn Phe Asn Lys Arg Phe Lys Pro 900 905 910 Asp Phe Pro Arg Thr Val Gln Arg Val Pro Tyr Gly Thr Tyr Ser Arg 915 920 925 Ile Glu Gly His Ala Pro 930 <210> 4 <211> 934 <212> PRT <213> Sorghum bicolor <400> 4 Met Phe Lys Ser Met Val Tyr Tyr Gly Asn Thr Ser Ile Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Glu Ala Ser Ala Gly Leu Ala Val Ala Ala 20 25 30 Trp Ala Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg 35 40 45 Cys Pro Pro Leu Ala Val Met His Thr Val Ala Val Gly Ala Arg Cys 50 55 60 Leu Val Met Glu Ser Arg Pro Pro Val Ala Ala Asp Val Ala Pro Met 65 70 75 80 Pro Leu Val Ala Met His Ala Ala Cys Leu Arg Asp Asn Lys Thr Ala 85 90 95 Val Val Pro Leu Gly Asp Gly Glu Leu His Leu Val Ala Met Thr Ser 100 105 110 Arg Arg Asn Leu Thr Asn His Ala Cys Phe Trp Gly Tyr Lys Leu Pro 115 120 125 Phe Gly Leu Tyr Asn Ser Cys Leu Thr Met Leu Asn Leu Arg Cys Leu 130 135 140 Gly Ile Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Ser 145 150 155 160 Arg Ser Phe Glu Asp Arg Ile Asp Gly Leu Gln Arg Lys Leu Ser Asn 165 170 175 Glu Thr Asp Pro Gln Arg Arg Asn Gly Met Leu Ser Glu Ile Lys Arg 180 185 190 Tyr Gln Asp Asp Lys Ser Ile Leu Lys Gln Tyr Ile Glu Gly Asp Gln 195 200 205 Val Tyr Asp Asp Gly Lys Val Tyr Lys Ala Gln Pro Glu Ile Val Pro 210 215 220 Pro Leu Ser Asp Asn Gln Gln Pro Met Thr Arg Pro Val Ile Arg Leu 225 230 235 240 Gln Asp Lys Asn Ile Ile Leu Thr Arg Ile Asn Pro Leu Ile Arg Asp 245 250 255 Thr Ser Val Leu Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser 260 265 270 Tyr Leu Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr 275 280 285 Met Ala Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro 290 295 300 Asp Ser Arg Leu Ile Asn Ser Val Gln Leu His Asp Arg Met Val Cys 305 310 315 320 Val Lys Ser Gly Leu Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly 325 330 335 Ser Cys His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val 340 345 350 Trp Asp Glu Lys Asp Gln Leu Arg Val His Val Val Pro Ala Phe Thr 355 360 365 Pro Tyr Tyr Ala Pro Gln Ala Glu Ala Asn Cys Ser Ile Pro Val Leu 370 375 380 Cys Val Ala Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys 385 390 395 400 Asp Phe Asp Glu Gly Leu Leu Pro Arg Ile Ser Asn Val His Tyr Glu 405 410 415 Asp Glu Val Asn Asp Ile Ser Ser Ala Pro Asp Val Gly Asn Tyr Leu 420 425 430 Ile Thr Glu Asp Glu Asn Ala Ala Leu Val Asn Gly Asn Arg Asp Ser 435 440 445 Leu Pro Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys 450 455 460 Glu Ala Asn Ala Gln Ala Phe His Gln Thr Ala Gly Asn Phe Val Met 465 470 475 480 Pro Val Ala Pro Ala Gln Asn Phe Val Ser Ser Ser Val Ala Pro Leu 485 490 495 Ala Pro Pro Leu Gly Val Met Pro Pro Thr Phe Ser Gln Pro Val Val 500 505 510 Gln Pro Gly Phe Ser Asp Ser Leu Gln Gly Ser Pro Ala Arg Glu Glu 515 520 525 Gly Glu Val Pro Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu 530 535 540 Leu Ile Leu Gln His Gly Gln Asp Ile Arg Asp Pro Thr Pro Pro Leu 545 550 555 560 Pro Ala Ile Pro Pro Val Gln Val Pro Val Pro Pro Val Gln Pro His 565 570 575 Gly Asn Trp Phe Pro Thr Glu Asp Gly Leu Asn Pro Ser Asn Leu Asn 580 585 590 Arg Gly Ser Ala Gly Phe Thr Val Glu Ser Asp Pro Met Leu Tyr Glu 595 600 605 Lys Lys Gln Pro Pro His Pro Ser Phe Phe His Gly Gly Asp Ser Pro 610 615 620 Met Ser Ser Asp Arg Phe Gly Tyr Gln Asn Gln Arg Phe Pro Ser Gln 625 630 635 640 Leu Pro His Thr Glu Asp His His Met Leu Gln Asn His Ala Pro Pro 645 650 655 Lys Tyr Arg Ser Phe Ser Gly Glu Glu Leu Ala Ala Arg His Val Pro 660 665 670 Ser Ser Gln Arg Asn Asn Gln Ile Glu Ser Gly Arg His Phe Ala Gln 675 680 685 Tyr Ala Gly Thr Ser Ala Gly Ile Leu Asp Gly Ile Ala Leu Lys Cys 690 695 700 Gly Ser Lys Val Glu Tyr Arg Ser Thr Leu Cys Asp Thr Ala Glu Leu 705 710 715 720 Gln Phe Ser Ile Glu Val Trp Ile Val Gly Glu Lys Val Gly Glu Gly 725 730 735 Ile Gly Arg Thr Arg Arg Glu Ala Gln His Lys Ala Ala Glu Met Ser 740 745 750 Leu Arg Asn Leu Ala Asn Lys Tyr Leu Ser Ser Asp Pro Asn Lys Leu 755 760 765 Thr Asp Met Lys Glu Asn Gly Phe Ser Gly Asn Arg Asn Val Phe Gly 770 775 780 Tyr Ser Gly Asn Thr Arg Asp Asp Met Leu Pro Leu Ser Ser Thr Ser 785 790 795 800 Glu Glu Ser Arg Phe Met Lys Met Glu Asn Asn Ser Arg Lys Thr Gly 805 810 815 Gly Ser Val Ala Ala Leu Lys Glu Leu Cys Thr Val Glu Gly Tyr Asn 820 825 830 Leu Val Phe Gln Glu Arg Pro Ser Pro Ala Asp Gly Leu Val Gly Lys 835 840 845 Glu Ser Tyr Ala Gln Val Glu Val Gly Gly Gln Ile Leu Gly Lys Gly 850 855 860 Val Gly Leu Thr Trp Glu Glu Ala Lys Leu Gln Ala Ala Asp Glu Ala 865 870 875 880 Leu Gly Thr Leu Arg Ser Met Leu Gly Gln Leu Ala His Lys Arg Ser 885 890 895 Gly Ser Pro Arg Ser Leu Ala Pro Asn Phe Asn Lys Arg Phe Lys Pro 900 905 910 Asp Phe Pro Arg Thr Val Gln Arg Val Pro Tyr Gly Thr Tyr Ser Arg 915 920 925 Ile Glu Gly His Val Pro 930 <210> 5 <211> 942 <212> PRT <213> Sorghum bicolor <400> 5 Met Ile Lys Ser Leu Val Tyr Tyr Gly Asn Thr Pro Val Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Gln Thr Asp Leu Ala Trp Ala Arg Glu Ile 20 25 30 Arg Val Asp Arg Leu Ser Pro Ala Ser Glu Arg Cys Pro Pro Leu Ala 35 40 45 Val Leu His Val Val Ala Ala Gly Ala Arg Cys Leu Val Met Glu Ser 50 55 60 Lys Ser Thr Ala Thr Ala His Glu Pro Pro Pro Pro Leu Val Thr Met 65 70 75 80 His Thr Thr Cys Leu Lys Asp Asn Lys Thr Ala Val Phe Pro Leu Gly 85 90 95 Ala Glu Glu Ile His Leu Val Ala Met Thr Ser Lys Arg Asn Met Pro 100 105 110 Asn Gly Ala Cys Phe Trp Gly Tyr Lys Val Pro Leu Gly Leu Tyr Asn 115 120 125 Ser Cys Leu Ser Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe Asp 130 135 140 Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr Arg Ser Phe Glu Asp 145 150 155 160 Arg Ile Asp Ala Ile Gln Arg Lys Leu Asn Asn Glu Ala Asp Pro Gln 165 170 175 Arg Ile Ser Gly Met Leu Ala Glu Ile Lys Arg Tyr Gln Glu Asp Lys 180 185 190 Ser Ile Leu Lys Gln Tyr Ile Glu Ser Asp Gln Val Thr Asp Gly Gly 195 200 205 Glu Leu Tyr Lys Val Gln Ser Glu Val Ile Pro Pro Leu Asp Asp Asn 210 215 220 His Gln Gln Pro Met Thr Arg Pro Ile Ile Arg Leu Gln Glu Lys Asn 225 230 235 240 Ile Ile Leu Thr Arg Ile Asn Pro Ser Ile Arg Asp Thr Ser Val Leu 245 250 255 Val Arg Leu Arg Pro Ala Trp Asp Asp Leu Arg Ser Tyr Leu Ile Ala 260 265 270 Arg Gly Arg Lys Arg Phe Glu Ile Tyr Val Cys Thr Met Ala Glu Arg 275 280 285 Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser Lys Leu 290 295 300 Ile Asn Ser Val Gln Leu Leu Asp Arg Leu Val Cys Val Lys Ser Gly 305 310 315 320 Ser Arg Lys Ser Leu Leu Asn Val Phe His Asp Gly Ser Cys His Pro 325 330 335 Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp Glu Trp 340 345 350 Asp Gln Arg Arg Val His Val Val Pro Ala Phe Ala Pro Tyr Tyr Ala 355 360 365 Pro Gln Ala Glu Ala Asn Phe Pro Ile Pro Val Leu Cys Val Ala Arg 370 375 380 Asn Val Ala Cys Asn Val Arg Ala Gly Phe Phe Lys Glu Phe Asp Glu 385 390 395 400 Gly Ile Ile Pro Arg Ile Thr Glu Val Cys Tyr Glu Asp Glu Leu Asp 405 410 415 Asp Ile Ala Ser Ala Pro Asp Val Cys Asn Tyr Phe Val Ser Glu Asp 420 425 430 Glu Asn Ala Ala Val Ser Asn Val Asn Lys Asn Pro Leu Ala Phe Asp 435 440 445 Gly Met Ala Asp Ala Glu Val Glu Lys Arg Met Lys Glu Ala Ser Ser 450 455 460 Ser Phe Gln Ser Ala Asn Pro Ile Thr Thr Asn Val Asp Val Met Ser 465 470 475 480 Val Ala Ala Asn Gln His Phe Gly Thr Pro Ile Ser Ser Ser Thr Pro 485 490 495 Val Ala Pro Pro Leu Gly Met Leu Leu Asn Asn Asp Gln Asp Pro Gln 500 505 510 Pro Pro Ser Leu Arg Trp Pro Val Ala Gln Ser Gly His Val Asp Ser 515 520 525 Ser Gln Gly Ser Pro Ala Arg Glu Glu Gly Glu Val Pro Glu Ser Glu 530 535 540 Leu Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln 545 550 555 560 Asp Thr Arg Asp Pro Ala Ala Pro Phe Pro Ala Gly Ser Pro Ala Gln 565 570 575 Val Ser Val Pro Pro Val Gln Ser His Glu Asn Trp Phe Cys Leu Glu 580 585 590 Asp Glu Met Asn Pro Arg Asn Leu Asn Lys Val Ser Thr Glu Phe His 595 600 605 Leu Glu Ser Asp Ser Val His Tyr Asp Lys Lys Gln Leu Gln His Thr 610 615 620 Ser Tyr Ile Pro Ile Gly Asp Asn Pro Met Ser Tyr Asp Arg Tyr Asp 625 630 635 640 Tyr Gln Asn Gln Arg Tyr Pro Ser Gln Pro Pro His Ser Glu Gly His 645 650 655 His Arg Phe His Asn His Ala Pro Thr Ala Tyr Arg Ser Phe Ser Gly 660 665 670 Glu Asp Met Ala Thr Trp Tyr Ala Pro Ser Gly Gln Arg Ser Ser His 675 680 685 Met Glu Ser Gly Arg His Phe Ala Arg Tyr Gly Gly Ile Pro Gly Val 690 695 700 Leu Glu Glu Ile Ala Leu Lys Cys Gly Phe Lys Val Glu Tyr Arg Ser 705 710 715 720 Thr Leu Cys Asp Thr Thr Glu Leu Gln Phe Ser Thr Glu Val Leu Ile 725 730 735 Phe Gly Glu Lys Val Gly Glu Gly Val Gly Lys Thr Arg Lys Glu Ala 740 745 750 Gln Trp Gln Ala Ala Asp Thr Ser Leu Arg Asn Leu Ala Asp Lys Phe 755 760 765 Leu Ser Trp Asp Pro Asp Lys Val Thr Val Leu Lys Glu Asn Asp Phe 770 775 780 Asn Arg His Pro Lys Ser His Arg Tyr Pro Gly Ser Asn Ile Tyr Asp 785 790 795 800 Thr Leu Pro Val Ala Ser Thr Ser Asp Glu Ser Arg Tyr Met Asn Asp 805 810 815 Arg Ile Asp Thr Leu Arg Lys Pro Gly Ala Ser Val Ala Ala Leu Lys 820 825 830 Glu Leu Cys Ala Val Glu Gly Tyr Asn Leu Asp Phe His Ala Gln Pro 835 840 845 Ser Ala Asp Gly Ser Val Gly Lys Glu Ile Arg Ala Gln Val Glu Ile 850 855 860 Gly Gly Lys Val Leu Gly Lys Gly Val Gly Val Thr Trp Glu Glu Ala 865 870 875 880 Lys Leu Gln Ala Ala Tyr Glu Ala Tyr Gly Thr Leu Lys Ser Met Leu 885 890 895 Gly Gln Phe Val Pro Arg Gln Ser Ala Ser Pro Arg Ser Met Val Pro 900 905 910 Asn Phe Asn Lys Arg Phe Asn Pro Asp Phe Ser Glu Ala Leu Gln Arg 915 920 925 Ile Pro Ser Gly Arg Tyr Ser Arg Asn Asp Ser Arg Phe Pro 930 935 940 <210> 6 <211> 948 <212> PRT <213> Triticum aestivum <400> 6 Met Ile Lys Ser Met Val Tyr Phe Gly His Ile Ser Ile Gly Glu Val 1 5 10 15 Glu Leu Ser Pro Lys Gly Glu Thr Asn Val Ala Ala Ala Pro Trp Val 20 25 30 Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg Cys Leu 35 40 45 Pro Leu Ala Val Leu His Thr Val Ser Ser Gly Ala Leu Cys Phe Val 50 55 60 Met Glu Ser Arg Pro Ser Pro Ala Thr Ala Asp Asn Glu Pro Pro Ser 65 70 75 80 Ser Leu Val Ala Met His Thr Ala Cys Leu Arg Asp Asn Lys Thr Ala 85 90 95 Val Phe Pro Leu Gly Ala Glu Glu Ile His Leu Val Ala Met Lys Pro 100 105 110 Lys Ser Asn Leu Pro Asn His Ala Cys Phe Trp Gly Tyr Lys Val Pro 115 120 125 Leu Gly Leu Tyr Ser Ser Cys Leu Ser Met Leu Asn Leu Arg Cys Leu 130 135 140 Gly Ile Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr 145 150 155 160 Arg Ser Phe Glu Asp Arg Ile Asp Ala Leu Gln Arg Lys Leu Ser Lys 165 170 175 Glu Thr Asp Pro Gln Arg Ile Ser Gly Met Leu Ala Glu Ile Lys Arg 180 185 190 Tyr Gln Glu Asp Arg Thr Met Leu Lys Gln Tyr Ile Asp Gly Asp Gln 195 200 205 Val Ile Asp Gly Gly Lys Met Tyr Lys Val Gln Ser Glu Val Val Pro 210 215 220 Pro Leu Ala Asp Asn His Gln Pro Met Ile Arg Pro Val Ile Arg Leu 225 230 235 240 Gln Asp Lys Ser Ile Ile Leu Thr Arg Ile Asn Pro Ser Ile Arg Asp 245 250 255 Thr Ser Val Leu Val Arg Leu Arg Pro Ala Trp Asp Asp Leu Arg Ser 260 265 270 Tyr Leu Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr 275 280 285 Met Ala Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro 290 295 300 Asp Ser Arg Leu Ile Asn Ser Val Gln Leu Pro His Arg Leu Val Cys 305 310 315 320 Val Lys Ser Gly Ser Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly 325 330 335 Ser Cys His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val 340 345 350 Trp Glu Glu Lys Asp Gln Cys Arg Val His Val Val Pro Ala Phe Ser 355 360 365 Pro Tyr Tyr Ala Pro Gln Ala Glu Ala Asn Phe Pro Ile Pro Val Leu 370 375 380 Cys Val Ala Arg Asn Val Ala Cys Asn Val Arg Gly Ser Phe Phe Lys 385 390 395 400 Glu Phe Asp Glu Gly Leu Leu Pro Ser Ile Ser Glu Val His Phe Asp 405 410 415 Asp Glu Leu Asp His Val Pro Ser Ser Pro Asp Val Gly Asn Tyr Leu 420 425 430 Ile Ser Glu Asp Glu Asn Ala Ala Ser Leu Asn Val Asn Lys Asp Pro 435 440 445 Met Ala Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys 450 455 460 Glu Ala Val Cys Ser Val Gln Ala Ala Asp Pro Val Thr Thr Asn Val 465 470 475 480 Asp Val Ile Ser Val Ala Ala Asn Gln Gln Phe Ala Thr Ser Ser Ser 485 490 495 Ile Pro Leu Ala Pro Pro Leu Gly Val Val Pro Leu Asn Asn Asp Gln 500 505 510 Gly Pro Gln Pro Pro Ser Val Ser Trp Pro Asp Ala Gln Ser Gly Met 515 520 525 Val Asp Pro Leu Gln Gly Ser Pro Ala Arg Glu Glu Gly Glu Val Pro 530 535 540 Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln 545 550 555 560 His Gly Gln Asp Thr Arg Asp Pro Thr Pro Pro Phe Ala Ala Glu Pro 565 570 575 Ser Val Gln Ala Ser Val Pro Pro Val Gln Ser Gln Gly Asn Trp Phe 580 585 590 Pro Val Glu Asp Glu Met Asp Pro Arg Asn Leu Asn Arg Thr Ser Thr 595 600 605 Asp Phe His Leu Glu Ser Asp Ala Val His Ser Asp Lys Ser Gln Pro 610 615 620 Pro His Gln Pro Tyr Phe Pro Ala Arg Asp Asn Pro Ile Phe Ser Asp 625 630 635 640 Arg Leu Asn His Gln Asn Gln Arg Tyr Ser Ser Gln Leu Pro His Ser 645 650 655 Glu Asp Arg Gln Met Leu Gln Asn Gln Ala Pro Thr Thr Tyr Arg Ser 660 665 670 Phe Ser Gly Glu Asp Met Ala Thr Gln Arg Phe His Pro Gly Asn Arg 675 680 685 Ser Ser Gln Met Glu Ser Gly Arg Gln Phe Val Gln Tyr Thr Glu Thr 690 695 700 Ser Gly Ala Val Leu Glu Glu Ile Ala Ala Lys Cys Gly Phe Lys Val 705 710 715 720 Glu Tyr Arg Ser Thr Leu Cys Asp Thr Thr Glu Leu Arg Phe Ser Ile 725 730 735 Gln Ile Trp Ile Val Gly Glu Lys Val Gly Glu Gly Met Gly Arg Thr 740 745 750 Arg Lys Glu Ala Gln Arg Gln Ala Ala Asn Ile Ser Leu Arg Asn Leu 755 760 765 Ala Asp Arg Phe Leu Ser Phe Asp Pro Asp Lys Met Thr Val Pro Val 770 775 780 Asp Asp Gly Phe Ser Ser Asn Pro Asn Ser Phe Lys Tyr Arg Gly Ile 785 790 795 800 Asp Gly Asp Asn Ile Val Pro Val Ala Ser Thr Ser Asp Gly Ser Arg 805 810 815 Tyr Met His Glu Arg Val Asp Asn Ser Thr Lys Ser Ala Gly Ser Val 820 825 830 Ala Ala Leu Lys Glu Leu Cys Thr Ala Glu Gly Tyr Asn Leu Val Phe 835 840 845 Gln Ala Gln Pro Ser Pro Ser Asp Ser Leu Arg Arg Glu Glu Val His 850 855 860 Ala Gln Ile Glu Ile Gly Gly Gln Ile Leu Gly Lys Gly Val Gly Val 865 870 875 880 Thr Trp Glu Glu Ala Lys Val Gln Ala Ala Asp Gly Ala Leu Gly Thr 885 890 895 Leu Arg Tyr Met Leu Gly Gln Arg Pro Gln Lys Arg Ser Gly Ser Pro 900 905 910 Arg Ser Phe Ala Ser Asn Tyr Asn Asn Lys Arg Tyr Lys Pro Asp Phe 915 920 925 Gln Pro Met Val Gln Arg Ile Pro Ser Gly Arg Tyr Ser Arg Asn Asp 930 935 940 Ser Arg Val Pro 945 <210> 7 <211> 948 <212> PRT <213> Triticum aestivum <400> 7 Met Ile Lys Ser Met Val Tyr Phe Gly His Ile Ser Ile Gly Glu Val 1 5 10 15 Glu Leu Trp Pro Lys Gly Glu Thr Asn Val Ala Ala Ala Pro Trp Val 20 25 30 Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg Cys Leu 35 40 45 Pro Leu Ala Val Leu His Thr Val Ser Ser Gly Ala Leu Cys Phe Val 50 55 60 Met Glu Ser Arg Pro Ser Pro Ala Thr Ala Asp Asp Glu Pro Pro Ser 65 70 75 80 Ser Leu Val Ala Met His Thr Ala Cys Leu Arg Asp Asn Lys Thr Ala 85 90 95 Val Phe Pro Leu Gly Ala Glu Glu Ile His Leu Val Ala Met Lys Pro 100 105 110 Lys Ser Ser Leu Pro Asn His Ala Cys Phe Trp Gly Tyr Lys Val Pro 115 120 125 Leu Gly Leu Tyr Asn Ser Cys Leu Ser Met Leu Asn Leu Arg Cys Leu 130 135 140 Gly Ile Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr 145 150 155 160 Arg Ser Phe Glu Asp Arg Ile Asp Ala Leu Gln Arg Lys Leu Ser Lys 165 170 175 Glu Ile Asp Pro Gln Arg Ile Ser Gly Met Leu Ala Glu Ile Lys Arg 180 185 190 Tyr Gln Glu Asp Arg Ser Met Leu Lys Gln Tyr Ile Asp Gly Asp Gln 195 200 205 Val Thr Asp Gly Gly Lys Val Tyr Lys Val Gln Ser Glu Val Val Pro 210 215 220 Pro Leu Ala Asp Asn His Gln Pro Met Ile Arg Pro Val Ile Arg Leu 225 230 235 240 Gln Glu Lys Ser Ile Ile Leu Thr Arg Ile Asn Pro Ser Ile Arg Asp 245 250 255 Thr Ser Val Leu Val Arg Leu Arg Pro Ala Trp Asp Asp Leu Arg Ser 260 265 270 Tyr Leu Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr 275 280 285 Methionine, Alanine, Glutamic acid, Arginine, Aspartic acid, Tyrosine, Alanine, Leucine, Glutamic acid, Methionine, Tryptophan, Arginine, Leucine, Leucine, Aspartic acid, Proline 290 295 300 Aspartic acid, Serine, Arginine, Leucine, Isoleucine, Asparagine, Serine, Valine, Glutamine, Leucine, Proline, Histidine, Arginine, Leucine, Valine, Cysteine 305 310 315 320 Valine, Lysine, Serine, Glycine, Serine, Lysine, Lysine, Serine, Leucine, Leucine, Asparagine, Valine, Phenylalanine, Histidine, Aspartic acid, Glycine 325 330 335 Serine, Cysteine, Histidine, Proline, Glycine, Methionine, Alanine, Leucine, Valine, Isoleucine, Aspartic acid, Aspartic acid, Arginine, Leucine, Lysine, Valine 340 345 350 Tryptophan, Glutamic acid, Glutamic acid, Lysine, Aspartic acid, Glutamine, Cysteine, Arginine, Valine, Histidine, Valine, Valine, Proline, Alanine, Phenylalanine, Serine 355 360 365 Proline, Tyrosine, Tyrosine, Alanine, Proline, Glutamine, Alanine, Glutamic acid, Alanine, Asparagine, Phenylalanine, Proline, Isoleucine, Proline, Valine, Leucine 370 375 380 Cysteine, Valine, Alanine, Arginine, Asparagine, Valine, Alanine, Cysteine, Asparagine, Valine, Arginine, Glycine, Serine, Phenylalanine, Phenylalanine, Lysine 385 390 395 400 Glutamic acid, Phenylalanine, Aspartic acid, Glutamic acid, Glycine, Leucine, Leucine, Proline, Serine, Isoleucine, Serine, Glutamic acid, Valine, Histidine, Phenylalanine, Aspartic acid 405 410 415 Aspartic acid, Glutamic acid, Leucine, Asparagine, Histidine, Valine, Proline, Serine, Serine, Proline, Aspartic acid, Valine, Glycine, Asparagine, Tyrosine, Leucine 420 425 430 Isoleucine, Proline, Glutamic acid, Aspartic acid, Glutamic acid, Asparagine, Alanine, Alanine, Serine, Leucine, Asparagine, Valine, Asparagine, Lysine, Aspartic acid, Glutamine 435 440 445 Met Ala Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys 450 455 460 Glu Ala Val Cys Ser Val Gln Ala Ala Asp Pro Val Thr Thr Asn Val 465 470 475 480 Asp Val Ile Pro Val Ala Ala Asn Gln Gln Phe Ala Thr Ser Ser Ser 485 490 495 Ile Pro Leu Ala Pro Pro Pro Gly Met Val Pro Leu Asn Asn Asp Gln 500 505 510 Gly Pro Gln Pro Pro Ser Val Ser Trp Pro Asp Ala Gln Ser Gly Met 515 520 525 Val Asp Pro Leu Gln Gly Ser Pro Ala Arg Glu Glu Gly Glu Val Pro 530 535 540 Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln 545 550 555 560 His Gly Gln Asp Thr Arg Asp Pro Thr Pro Pro Phe Ala Ala Glu Pro 565 570 575 Ser Val Gln Ala Ser Val Pro Pro Val Gln Ser Gln Gly Asn Trp Phe 580 585 590 Pro Val Glu Asp Glu Met Asp Pro Arg Asn Leu Asn Arg Thr Ser Thr 595 600 605 Asp Phe His Leu Glu Ser Asp Ala Val His Ser Asp Lys Ser Gln Pro 610 615 620 Pro His Gln Pro Tyr Phe Pro Ala His Asp Asn Pro Ile Phe Ser Asp 625 630 635 640 Arg Leu Asn His Gln Asn Gln Arg Tyr Ser Ser Gln Leu Pro His Ser 645 650 655 Glu Asp Arg Gln Met Leu Gln Asn Gln Ala Pro Thr Thr Tyr Arg Ser 660 665 670 Phe Ser Gly Glu Asp Met Ala Thr Gln Arg Phe His Pro Gly Asn Arg 675 680 685 Ser Ser Gln Met Glu Ser Gly Arg Gln Phe Val Gln Tyr Thr Glu Thr 690 695 700 Ser Gly Ala Val Leu Glu Glu Ile Ala Ala Lys Cys Gly Phe Lys Val 705 710 715 720 Glu Tyr Arg Ser Thr Leu Cys Asp Thr Thr Glu Leu Arg Phe Ser Ile 725 730 735 Gln Ile Trp Ile Val Gly Glu Lys Val Gly Glu Gly Met Gly Arg Thr 740 745 750 Arg Lys Glu Ala Gln Arg Gln Ala Ala Asn Ile Ser Leu Arg Asn Leu 755 760 765 Ala Asp Arg Phe Leu Ser Phe Asp Pro Asp Lys Met Thr Val Pro Met 770 775 780 Asp Asp Gly Phe Ser Ser Asn Pro Asn Ser Phe Lys Tyr Arg Gly Ile 785 790 795 800 Asp Gly Asp Asn Ile Val Pro Val Ala Ser Thr Ser Asp Gly Ser Arg 805 810 815 Tyr Met His Glu Arg Val Asp Asn Ser Thr Lys Ser Ala Gly Ser Val 820 825 830 Ala Ala Leu Lys Glu Leu Cys Thr Ala Glu Gly Tyr Asn Leu Val Phe 835 840 845 Gln Ala Gln Pro Ser Pro Leu Asp Ser Leu Arg Arg Glu Glu Val His 850 855 860 Ala Gln Ile Glu Ile Gly Gly Gln Ile Leu Gly Lys Gly Val Gly Val 865 870 875 880 Thr Trp Glu Glu Ala Lys Glu Gln Ala Ala Asp Gly Ala Leu Arg Thr 885 890 895 Leu Arg Tyr Met Leu Gly Gln Arg Pro Gln Lys Arg Pro Gly Ser Pro 900 905 910 Arg Ser Phe Ala Ser Asn Tyr Asn Asn Lys Arg Tyr Lys Pro Asp Phe 915 920 925 Gln Pro Met Val Gln Arg Ile Pro Ser Gly Arg Tyr Ser Arg Asn Asp 930 935 940 Ser Arg Val Pro 945 <210> 8 <211> 947 <212> PRT <213> Triticum aestivum <400> 8 Met Ile Lys Ser Met Val Tyr Phe Gly His Ile Ser Ile Gly Glu Val 1 5 10 15 Glu Leu Trp Pro Lys Gly Glu Thr Asn Val Ala Ala Ala Pro Trp Val 20 25 30 Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg Cys Leu 35 40 45 Pro Leu Ala Val Leu His Thr Val Ser Ser Gly Ala Leu Cys Phe Val 50 55 60 Met Glu Ser Arg Pro Ser Pro Ala Thr Ala Asp Asp Glu Pro Pro Ser 65 70 75 80 Ser Leu Val Ala Met His Thr Ala Cys Leu Arg Asp Asn Lys Thr Ala 85 90 95 Val Phe Pro Leu Gly Ala Glu Glu Ile His Leu Val Ala Met Lys Pro 100 105 110 Lys Ser Asn Leu Pro Asn His Ala Cys Phe Trp Gly Tyr Lys Val Pro 115 120 125 Leu Gly Leu Tyr Asn Ser Cys Leu Ser Met Leu Asn Leu Arg Cys Leu 130 135 140 Gly Ile Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr 145 150 155 160 Arg Ser Phe Glu Asp Arg Ile Asp Ala Leu Gln Arg Lys Leu Ser Lys 165 170 175 Glu Thr Asp Pro Gln Arg Ile Ser Gly Met Leu Ala Glu Ile Lys Arg 180 185 190 Tyr Gln Glu Asp Arg Thr Met Leu Lys Gln Tyr Ile Asp Gly Asp Gln 195 200 205 Val Ile Asp Gly Gly Lys Met Tyr Lys Val Gln Ser Glu Val Val Pro 210 215 220 Pro Leu Ala Asp Asn His Gln Pro Met Ile Arg Pro Val Ile Arg Leu 225 230 235 240 Gln Glu Lys Ser Ile Ile Leu Thr Arg Ile Asn Pro Ser Ile Arg Asp 245 250 255 Thr Ser Val Leu Val Arg Leu Arg Pro Ala Trp Asp Asp Leu Arg Ser 260 265 270 Tyr Leu Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr 275 280 285 Met Ala Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro 290 295 300 Asp Ser Arg Leu Ile Asn Ser Val Gln Leu Pro His Arg Leu Val Cys 305 310 315 320 Val Lys Ser Gly Phe Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly 325 330 335 Ser Cys His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val 340 345 350 Trp Glu Glu Lys Asp Gln Cys Arg Val His Val Val Pro Ala Phe Ser 355 360 365 Pro Tyr Tyr Ala Pro Gln Ala Glu Ala Asn Phe Pro Ile Pro Val Leu 370 375 380 Cys Val Ala Arg Asn Val Ala Cys Asn Val Arg Gly Ser Phe Phe Lys 385 390 395 400 Glu Phe Asp Glu Gly Leu Leu Pro Ser Ile Ser Glu Val His Phe Asp 405 410 415 Asp Glu Leu Asp His Val Pro Ser Ser Pro Asp Val Gly Asn Tyr Leu 420 425 430 Ile Ser Glu Asp Glu Asn Ala Ala Ser Leu Asn Val Asn Lys Asp Pro 435 440 445 Met Ala Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys 450 455 460 Glu Ala Val Cys Ser Val Gln Ala Ala Asp Pro Val Thr Thr Asn Val 465 470 475 480 Asp Val Ile Ser Ala Ala Ala Asn Gln Gln Phe Ala Thr Ser Ser Ser 485 490 495 Ile Pro Leu Ala Pro Pro Leu Gly Met Val Pro Leu Asn Asn Asp Gln 500 505 510 Gly Pro Gln Pro Pro Ser Val Ser Trp Ser Asp Ala Gln Ser Gly Met 515 520 525 Val Asp Pro Leu Gln Gly Ser Pro Ala Arg Glu Glu Gly Glu Val Pro 530 535 540 Glu Ser Glu Leu Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln 545 550 555 560 His Gly Gln Asp Thr Arg Asp Pro Thr Pro Pro Phe Ala Ala Glu Pro 565 570 575 Ser Val Gln Ala Ser Val Pro Pro Val Gln Ser Gln Gly Asn Trp Phe 580 585 590 Pro Val Glu Asp Glu Met Asp Pro Arg Asn Leu Asn Arg Thr Ser Thr 595 600 605 Asp Phe His Val Glu Ser Asp Ala Val His Ser Asp Lys Ser Gln Pro 610 615 620 Pro His Gln Pro Tyr Phe Pro Ala Arg Asp Asn Pro Val Phe Ser Asp 625 630 635 640 Arg Phe Asn His Gln Asn Gln Arg Tyr Ser Ser Gln Leu Pro His Ser 645 650 655 Glu Asp Arg Gln Met Leu Gln Asn Gln Ala Pro Thr Thr Tyr Arg Ser 660 665 670 Phe Ser Gly Glu Asp Met Ala Thr Gln Arg Phe His Pro Gly Asn Arg 675 680 685 Ser Ser Gln Met Glu Ser Gly Arg Gln Phe Val Gln Tyr Thr Glu Thr 690 695 700 Ser Gly Ala Val Leu Glu Glu Ile Ala Ala Lys Cys Gly Phe Lys Val 705 710 715 720 Glu Tyr Arg Ser Thr Leu Cys Asp Thr Thr Glu Leu Arg Phe Ser Ile 725 730 735 Gln Ile Trp Ile Val Gly Glu Lys Val Gly Glu Gly Met Gly Arg Thr 740 745 750 Arg Lys Glu Ala Gln Arg Gln Ala Ala Asn Ile Ser Leu Arg Asn Leu 755 760 765 Ala Asp Lys Phe Leu Ser Phe Asp Pro Asp Lys Met Thr Val Pro Met 770 775 780 Asp Asp Gly Phe Ser Ser Asn Pro Asn Ser Phe Lys Tyr Thr Gly Ile 785 790 795 800 Asp Gly Asp Asn Ile Val Pro Val Ala Ser Thr Ser Asp Gly Ser Arg 805 810 815 Tyr Met His Glu Arg Val Asp Asn Ser Thr Lys Ser Ala Gly Ser Val 820 825 830 Ala Ala Leu Lys Glu Leu Cys Thr Ala Glu Gly Tyr Asn Leu Val Phe 835 840 845 Gln Ala Gln Pro Ser Pro Leu Asp Ser Leu Thr Arg Lys Glu Val His 850 855 860 Ala Gln Ile Glu Ile Gly Gly Gln Ile Leu Gly Lys Gly Val Gly Val 865 870 875 880 Thr Trp Glu Glu Ala Lys Val Gln Ala Ala Asp Gly Ala Leu Gly Thr 885 890 895 Leu Arg Tyr Met Leu Gly Gln Arg Pro Gln Lys Arg Ser Gly Ser Pro 900 905 910 Arg Ser Phe Ala Ser Asn Tyr Asn Lys Arg Tyr Lys Pro Asp Phe Gln 915 920 925 Pro Met Ala Gln Arg Ile Pro Ser Gly Arg Tyr Ser Arg Asn Asp Ser 930 935 940 Arg Val Pro 945 <210> 9 <211> 934 <212> PRT <213> Triticum aestivum <400> 9 Met Ile Lys Ser Met Val Tyr Tyr Gly Asn Thr Ser Ile Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Asp Thr Asn Leu Gly Ala Ala Ala Trp Ala 20 25 30 Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg Cys Leu 35 40 45 Pro Leu Ala Val Met His Thr Val Ala Val Gly Ala Arg Cys Leu Val 50 55 60 Met Glu Ser Arg Pro Pro Lys Ala Ala Asp Glu Pro Pro Pro Pro Leu 65 70 75 80 Val Ala Met His Ala Ala Cys Leu Arg Asp Asn Lys Thr Ala Val Val 85 90 95 Pro Leu Gly Glu Glu Glu Leu His Leu Val Ala Met Thr Ser Gly Arg 100 105 110 Asn Leu Thr Asn His Ala Cys Phe Trp Gly Tyr Lys Val Pro Phe Gly 115 120 125 Leu Tyr Asn Ser Cys Leu Thr Met Leu Asn Leu Arg Cys Leu Gly Ile 130 135 140 Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr Arg Ser 145 150 155 160 Phe Glu Asp Arg Ile Asp Ser Leu Gln Arg Lys Leu Ser Asn Glu Thr 165 170 175 Asp Pro Gln Arg Met Asn Gly Met Leu Ala Glu Ile Lys Arg Tyr Gln 180 185 190 Asp Asp Arg Ser Ile Leu Lys Gln Tyr Ile Glu Gly Asp Gln Val Tyr 195 200 205 Asp Asp Gly Lys Met Tyr Lys Val Gln Pro Glu Ile Val Pro Pro Leu 210 215 220 Ser Asp Asn His Gln Ser Leu Thr Arg Pro Val Ile Arg Leu Gln Glu 225 230 235 240 Lys Asn Ile Ile Leu Thr Arg Ile Asn Pro Leu Ile Arg Asp Thr Ser 245 250 255 Val Leu Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu 260 265 270 Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala 275 280 285 Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser 290 295 300 Arg Leu Ile Asn Ser Val Gln Leu Ser Asp Arg Met Val Cys Val Lys 305 310 315 320 Ser Gly Leu Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly Ser Cys 325 330 335 His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp 340 345 350 Glu Lys Asp Gln Ser Arg Val His Val Val Pro Ala Phe Thr Pro Tyr 355 360 365 Tyr Ala Pro Gln Ala Glu Ala Asn Cys Ser Ile Pro Val Leu Cys Val 370 375 380 Ala Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Asp Phe 385 390 395 400 Asp Glu Gly Leu Leu Pro Arg Ile Thr Ser Val Leu Tyr Glu Asp Glu 405 410 415 Ile Gln Asp Ile Ser Ser Ala Pro Asp Val Gly Asn Tyr Leu Ile Ser 420 425 430 Glu Asp Glu Asn Val Ala Val Val Asn Gly Asn Arg Asp Ser Leu Ala 435 440 445 Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys Glu Ala 450 455 460 Ser Gly Ser Gly Ser Val Leu Asn Pro Thr Met Ala Asn Met Val Met 465 470 475 480 Pro Val Ala Pro Ser Gln Ser Phe Ala Pro Ser Ser Val Ala Pro Phe 485 490 495 Ala Pro Pro Leu Gly Met Met Pro Leu Ser Asn Asn Gln Val Pro Pro 500 505 510 Pro Ala Phe Ser Gln Pro Val Val Gln Pro Val Val Leu Asp Pro Leu 515 520 525 Gln Ala Ser Pro Gly Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu 530 535 540 Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp 545 550 555 560 Thr Arg Asp Pro Thr Pro Pro Leu Pro Ala Val Pro Pro Ala Gln Val 565 570 575 Ser Val Pro Pro Val Gln Ser His Gly Asn Trp Phe Pro Ile Glu Asp 580 585 590 Gly Ile Gly Met Asn Ser Asn Asn Leu Asn Met Gly Ser Ala Gly Phe 595 600 605 Pro Ser Glu Ser Asp Thr Met His Tyr Asp Lys Lys Gln Pro Pro Gln 610 615 620 Pro Ser Tyr Phe His Gly Gly Asp Asn Asn Pro Val Pro Ser Asp Arg 625 630 635 640 Phe Ser Tyr Gln Ser Gln Arg Phe Pro Ser Gln Val Thr His Thr Glu 645 650 655 Asp His Arg Met Leu Gln Asn His Ala Pro Pro Arg Tyr Arg Ser Phe 660 665 670 Pro Gly Gln Arg Asn Asn Leu Ile Glu Ser Gly Gln Ser Tyr Ala Arg 675 680 685 Asn Val Gly Ser Val Gly Ile Leu Glu Glu Ile Ala Leu Lys Ser Gly 690 695 700 Ser Lys Val Glu Tyr Arg Ser Thr Leu Cys Asp Thr Ala Glu Leu Gln 705 710 715 720 Phe Ser Ile Glu Ile Trp Ile Val Gly Glu Lys Val Gly Glu Gly Ile 725 730 735 Gly Ser Ser Arg Lys Glu Ala Gln Arg Gln Ala Ala Glu Ile Ser Leu 740 745 750 Arg Asn Leu Ala Asn Lys Tyr Leu Leu Ser Asp Pro Asn Lys Met Thr 755 760 765 Asp Val Asn Glu Asp Gly Phe Gly Ser Asn Pro Asn Phe Phe Gly Tyr 770 775 780 Ser Glu Asn Thr Arg Asn Asp Ile Leu Ser Val Ala Ser Thr Ser Glu 785 790 795 800 Glu Ser Arg Phe Thr Lys Thr Gly Glu Asn Asn Ser Arg Ile Thr Gly 805 810 815 Gly Ser Ile Ala Ala Leu Lys Gln Leu Cys Thr Val Glu Gly Tyr Asn 820 825 830 Leu Val Phe Gln Ala Arg Pro Ser Pro Leu Asp Gly Ser Gly Gly Lys 835 840 845 Glu Thr Tyr Ala Gln Val Glu Val Gly Gly Gln Thr Leu Gly Lys Gly 850 855 860 Val Gly Ile Thr Trp Glu Glu Ala Lys Leu Gln Ala Ala Asp Glu Ala 865 870 875 880 Leu Gly Thr Leu Arg Ser Met Leu Gly Gln Leu Ala Gln Lys Arg Ser 885 890 895 Ser Ser Pro Arg Ser Leu Ala Pro Asn Tyr Asn Lys Arg Phe Lys Pro 900 905 910 Asp Phe Pro Arg Ala Val Gln Arg Pro Pro Tyr Gly Arg Tyr Ser Arg 915 920 925 Ile Glu Gly His Val Pro 930 <210> 10 <211> 935 <212> PRT <213> Triticum aestivum <400> 10 Met Ile Lys Ser Met Val Tyr Tyr Gly Asn Thr Ser Ile Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Asp Thr Asn Leu Gly Ala Ala Ala Trp Ala 20 25 30 Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg Cys Leu 35 40 45 Pro Leu Ala Val Met His Thr Val Ala Val Gly Ala Arg Cys Leu Val 50 55 60 Met Glu Ser Arg Pro Pro Lys Ala Ala Asp Glu Pro Pro Pro Pro Leu 65 70 75 80 Val Ala Met His Ala Ala Cys Leu Arg Asp Asn Lys Thr Ala Val Val 85 90 95 Pro Leu Gly Glu Glu Glu Leu His Leu Val Ala Met Thr Ser Gly Arg 100 105 110 Asn Leu Thr Asn His Ala Cys Phe Trp Gly Tyr Lys Val Pro Phe Gly 115 120 125 Leu Tyr Asn Ser Cys Leu Thr Met Leu Asn Leu Arg Cys Leu Gly Ile 130 135 140 Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr Arg Thr 145 150 155 160 Phe Glu Asp Arg Ile Asp Ser Leu Gln Arg Lys Leu Ser Asn Glu Thr 165 170 175 Asp Pro Gln Arg Met Asn Gly Met Leu Ala Glu Ile Lys Arg Tyr Gln 180 185 190 Asp Asp Arg Ser Ile Leu Lys Gln Tyr Ile Glu Gly Asp Gln Val Tyr 195 200 205 Asp Asp Gly Lys Met Tyr Lys Val Gln Pro Glu Ile Val Pro Pro Leu 210 215 220 Ser Asp Asn His Gln Ser Leu Thr Arg Pro Val Ile Arg Leu Gln Glu 225 230 235 240 Lys Asn Ile Ile Leu Thr Arg Ile Asn Pro Leu Ile Arg Asp Thr Ser 245 250 255 Val Leu Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu 260 265 270 Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala 275 280 285 Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser 290 295 300 Arg Leu Ile Asn Ser Val Gln Leu Ser Asp Arg Met Val Cys Val Lys 305 310 315 320 Ser Gly Leu Lys Lys Ser Leu Leu Asn Val Phe His Asp Arg Ser Cys 325 330 335 His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp 340 345 350 Glu Lys Asp Gln Ser Arg Val His Val Val Pro Ala Phe Thr Pro Tyr 355 360 365 Tyr Ala Pro Gln Ala Glu Ala Asn Cys Ser Ile Pro Val Leu Cys Val 370 375 380 Ala Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Asp Phe 385 390 395 400 Asp Glu Gly Leu Leu Pro Arg Ile Thr Ser Val Leu Tyr Glu Asp Glu 405 410 415 Ile Gln Asp Ile Ser Ser Ala Pro Asp Val Gly Asn Tyr Leu Ile Ser 420 425 430 Glu Asp Glu Asn Val Ala Val Val Asn Gly Asn Arg Asp Ser Leu Ala 435 440 445 Phe Asp Gly Met Ala Asp Ala Glu Gly Glu Arg Arg Met Lys Glu Ala 450 455 460 Ser Gly Ser Cys Ser Val Leu Asn Pro Thr Met Ala Asn Met Val Met 465 470 475 480 Pro Val Ala Pro Ser Gln Ser Phe Val Pro Ser Ser Val Ala Pro Phe 485 490 495 Ala Pro Pro Leu Gly Met Met Pro Leu Ser Asn Asn Gln Val Pro Pro 500 505 510 Pro Val Phe Ser Gln Pro Val Val Gln Pro Val Val Leu Asp Pro Leu 515 520 525 Gln Ala Ser Pro Gly Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu 530 535 540 Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp 545 550 555 560 Thr Arg Asp Pro Thr Pro Pro Leu Pro Ala Val Pro Pro Ala Gln Val 565 570 575 Ser Val Pro Pro Val Gln Ser His Gly Asn Trp Phe Pro Val Glu Asp 580 585 590 Gly Ile Gly Met Asn Ser Asn Asn Leu Asn Arg Gly Ser Ala Gly Phe 595 600 605 Pro Ser Glu Ser Asp Thr Met His Tyr Asp Lys Lys Gln Pro Pro Gln 610 615 620 Pro Ser Tyr Phe His Gly Gly Asp Asn Asn Pro Val Ser Ser Asp Arg 625 630 635 640 Phe Ser Tyr Gln Ser Gln Arg Phe Pro Ser Gln Val Thr His Thr Glu 645 650 655 Asp His Arg Met Leu Gln Asn His Ala Pro Pro Arg Tyr Arg Ser Phe 660 665 670 Pro Gly Gln Arg Asp Asn Leu Ile Glu Ser Gly Gln Ser Tyr Ala Arg 675 680 685 Asn Val Gly Thr Ser Val Gly Ile Leu Glu Glu Ile Ala Leu Lys Ser 690 695 700 Gly Ser Lys Val Glu Tyr Arg Ser Lys Leu Cys Asp Thr Ala Glu Leu 705 710 715 720 Gln Phe Ser Ile Glu Val Trp Ile Val Gly Glu Lys Val Gly Glu Gly 725 730 735 Ile Gly Ser Ser Arg Lys Glu Ala Gln Arg Gln Ala Ala Glu Ile Ser 740 745 750 Leu Arg Asn Leu Ala Asn Lys Tyr Leu Leu Ser Asp Pro Asn Lys Met 755 760 765 Thr Asp Val Asn Glu Asp Gly Phe Gly Ser Asn Pro Asn Phe Phe Gly 770 775 780 Tyr Ser Glu Asn Thr Arg Asn Asp Ile Leu Pro Val Ala Ser Thr Ser 785 790 795 800 Glu Glu Ser Arg Phe Thr Lys Thr Gly Glu Asn Asn Ser Arg Ile Thr 805 810 815 Gly Gly Ser Ile Ala Ala Leu Lys Gln Leu Cys Thr Val Glu Gly Tyr 820 825 830 Asn Leu Val Phe Gln Ala Arg Pro Ser Pro Leu Asp Gly Ser Gly Gly 835 840 845 Lys Glu Thr Tyr Ala Gln Val Glu Val Gly Gly Gln Thr Leu Gly Lys 850 855 860 Gly Val Gly Ile Thr Trp Glu Glu Ala Lys Leu Gln Ala Ala Asp Glu 865 870 875 880 Ala Leu Gly Thr Leu Arg Ser Met Leu Gly Gln Leu Ala Gln Lys Arg 885 890 895 Ser Ser Ser Pro Arg Ser Leu Ala Pro Asn Tyr Asn Lys Arg Phe Lys 900 905 910 Pro Asp Phe Pro Arg Ala Val Gln Arg Pro Pro Tyr Gly Arg Tyr Ser 915 920 925 Arg Ile Glu Gly His Val Pro 930 935 <210> 11 <211> 935 <212> PRT <213> Triticum aestivum <400> 11 Met Ile Lys Ser Met Val Tyr Tyr Gly Asn Thr Ser Ile Gly Glu Val 1 5 10 15 Glu Val Trp Pro Lys Gly Asp Thr Asn Leu Gly Ala Ala Ala Trp Ala 20 25 30 Arg Glu Ile Arg Val Asp Arg Leu Ser Pro Pro Ser Glu Arg Cys Leu 35 40 45 Pro Leu Ala Val Met His Thr Val Ala Val Gly Ala Arg Cys Leu Asp 50 55 60 Met Glu Ser Arg Pro Pro Lys Ala Ala Asp Glu Pro Pro Pro Pro Leu 65 70 75 80 Val Ala Met His Ala Ala Cys Leu Arg Asp Asn Lys Thr Ala Val Val 85 90 95 Pro Leu Gly Glu Glu Glu Leu His Leu Val Ala Met Thr Ser Gly Arg 100 105 110 Asn Leu Thr Asn His Ala Cys Phe Trp Gly Tyr Lys Val Pro Phe Gly 115 120 125 Leu Tyr Asn Ser Cys Leu Thr Met Leu Asn Leu Arg Cys Leu Gly Ile 130 135 140 Val Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Thr Arg Ser 145 150 155 160 Phe Glu Asp Arg Ile Asp Ser Leu Gln Arg Lys Leu Ser Asn Glu Thr 165 170 175 Asp Pro Gln Arg Met Asn Gly Met Leu Ala Glu Ile Lys Arg Tyr Gln 180 185 190 Asp Asp Arg Ser Ile Leu Lys Gln Tyr Ile Glu Gly Asp Gln Val Tyr 195 200 205 Asp Asp Gly Lys Met Tyr Lys Val Gln Pro Glu Ile Val Pro Pro Leu 210 215 220 Ser Asp Asn His Gln Ser Leu Thr Arg Pro Val Ile Arg Leu Gln Glu 225 230 235 240 Lys Asn Ile Ile Leu Thr Arg Ile Asn Pro Leu Ile Arg Asp Thr Ser 245 250 255 Val Leu Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu 260 265 270 Ile Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala 275 280 285 Glu Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser 290 295 300 Arg Leu Ile Asn Ser Val Gln Leu Ser Asp Arg Met Val Cys Val Lys 305 310 315 320 Ser Gly Leu Lys Lys Ser Leu Leu Asn Val Phe His Asp Gly Ser Cys 325 330 335 His Pro Gly Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp 340 345 350 Glu Lys Asp Gln Ser Arg Val His Val Val Pro Ala Phe Thr Pro Tyr 355 360 365 Tyr Ala Pro Gln Ala Glu Ala Asn Cys Ser Ile Pro Val Leu Cys Val 370 375 380 Ala Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Asp Phe 385 390 395 400 Asp Glu Gly Leu Leu Pro Arg Ile Thr Ser Val Leu Tyr Glu Asp Glu 405 410 415 Ile Gln Asp Ile Ser Ser Ala Pro Asp Val Gly Asn Tyr Leu Ile Ser 420 425 430 Glu Asp Glu Asn Val Ala Val Val Asn Gly Asn Arg Asp Ser Leu Ala 435 440 445 Phe Asp Gly Met Ala Asp Ala Glu Val Glu Arg Arg Met Lys Glu Ala 450 455 460 Ser Gly Ser Gly Ser Val Leu Asn Pro Thr Met Ala Asn Met Val Met 465 470 475 480 Pro Val Ala Pro Ser Gln Ser Phe Val Pro Ser Ser Val Ala Pro Phe 485 490 495 Ala Pro Pro Leu Gly Met Met Pro Leu Ser Asn Asn Gln Val Pro Pro 500 505 510 Pro Ala Leu Ser Gln Pro Val Val Gln Pro Val Val Leu Asp Pro Leu 515 520 525 Gln Ala Ser Pro Gly Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu 530 535 540 Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp 545 550 555 560 Thr Arg Asp Pro Thr Pro Pro Leu Pro Ala Val Pro Pro Ala Gln Val 565 570 575 Ser Val Pro Pro Val Gln Ser His Gly Asn Trp Phe Pro Ile Glu Asp 580 585 590 Gly Ile Gly Met Asn Ser Asn Asn Leu Asn Met Gly Ser Ala Gly Phe 595 600 605 Pro Ser Glu Ser Asp Thr Met His Tyr Asp Lys Lys Gln Pro Pro Gln 610 615 620 Pro Ser Tyr Phe His Gly Gly Asp Asn Asn Pro Val Ser Ser Gly Arg 625 630 635 640 Phe Ser Tyr Gln Ser Gln Arg Phe Pro Ser Gln Val Thr His Thr Glu 645 650 655 Asp His Arg Met Leu Gln Asn His Ala Pro Pro Arg Tyr Arg Ser Phe 660 665 670 Pro Gly Gln Arg Asn Asn Leu Ile Glu Ser Gly Gln Ser Tyr Ala Arg 675 680 685 Asn Val Gly Thr Ser Val Gly Ile Leu Glu Glu Ile Ala Leu Lys Ser 690 695 700 Gly Cys Lys Val Glu Tyr Arg Ser Thr Leu Cys Asp Thr Ala Glu Leu 705 710 715 720 Gln Phe Ser Ile Glu Val Trp Ile Val Gly Glu Lys Val Gly Glu Gly 725 730 735 Ile Gly Ser Ser Arg Lys Glu Ala Gln Arg Gln Ala Ala Glu Ile Ser 740 745 750 Leu Arg Asn Leu Ala Asn Lys Tyr Leu Leu Ser Asp Pro Asn Lys Met 755 760 765 Thr Asp Gly Asn Glu Asp Gly Phe Gly Ser Asn Pro Asn Phe Phe Gly 770 775 780 Tyr Ser Glu Asn Thr Arg Asn Asp Ile Leu Pro Val Ala Ser Thr Ser 785 790 795 800 Glu Glu Ser Arg Phe Thr Lys Thr Gly Glu Asn Asn Ser Arg Ile Thr 805 810 815 Gly Gly Ser Ile Ala Ala Leu Lys Gln Leu Cys Thr Val Glu Gly Tyr 820 825 830 Asn Leu Val Phe Gln Ala Arg Pro Ser Pro Leu Asp Gly Ser Gly Gly 835 840 845 Lys Glu Thr Tyr Ala Gln Val Glu Val Gly Gly Gln Thr Leu Gly Lys 850 855 860 Gly Val Gly Ile Thr Trp Glu Glu Ala Lys Leu Gln Ala Ala Asp Glu 865 870 875 880 Ala Leu Gly Thr Leu Arg Ser Met Leu Gly Gln Leu Ala Gln Lys Arg 885 890 895 Ser Ser Ser Pro Arg Ser Leu Val Pro Asn Tyr Asn Lys Arg Phe Lys 900 905 910 Pro Asp Phe Pro Arg Ala Val Gln Arg Pro Pro Tyr Gly Arg Tyr Ser 915 920 925 Arg Ile Glu Gly His Val Pro 930 935 <210> 12 <211> 956 <212> PRT <213> Glycine max <400> 12 Met Arg Met Tyr Lys Ser Val Val Tyr Gln Gly Glu Val Val Val Gly 1 5 10 15 Glu Val Asp Val Tyr Pro Glu Glu Asn Asn Asn Tyr Lys Asn Phe His 20 25 30 Val Lys Glu Ile Arg Ile Ser His Phe Ser Gln Pro Ser Glu Arg Cys 35 40 45 Pro Pro Leu Ala Val Leu His Thr Val Thr Ser Cys Gly Val Cys Phe 50 55 60 Lys Met Glu Ser Lys Thr Gln Gln Gln Asp Gly Leu Phe Gln Leu His 65 70 75 80 Ser Leu Cys Ile Arg Glu Asn Lys Thr Ala Val Met Pro Leu Gly Gly 85 90 95 Glu Glu Ile His Leu Val Ala Met His Ser Arg Asn Val Asp Arg Pro 100 105 110 Cys Phe Trp Gly Phe Ile Val Ala Leu Gly Leu Tyr Asp Ser Cys Leu 115 120 125 Val Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe Asp Leu Asp Glu 130 135 140 Thr Leu Ile Val Ala Asn Thr Met Arg Ser Phe Glu Asp Arg Ile Asp 145 150 155 160 Ala Leu Gln Arg Lys Ile Asn Ser Glu Val Asp Pro Gln Arg Ile Ser 165 170 175 Gly Met Gln Ala Glu Val Lys Arg Tyr Gln Asp Asp Lys Asn Ile Leu 180 185 190 Lys Gln Tyr Ala Glu Asn Asp Gln Val Val Asp Asn Gly Arg Val Ile 195 200 205 Lys Val Gln Ser Glu Ile Val Pro Ala Leu Ser Asp Ser His Gln Pro 210 215 220 Ile Val Arg Pro Leu Ile Arg Leu Gln Asp Lys Asn Ile Ile Leu Thr 225 230 235 240 Arg Ile Asn Pro Gln Ile Arg Asp Thr Ser Val Leu Val Arg Leu Arg 245 250 255 Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu Thr Ala Arg Gly Arg Lys 260 265 270 Arg Phe Glu Val Tyr Val Cys Thr Met Ala Glu Arg Asp Tyr Ala Leu 275 280 285 Glu Met Trp Arg Leu Leu Asp Pro Asp Ser Asn Leu Ile Asn Ser Lys 290 295 300 Glu Leu Leu Gly Arg Ile Val Cys Val Lys Ser Gly Leu Lys Lys Ser 305 310 315 320 Leu Phe Asn Val Phe Gln Asp Gly Leu Cys His Pro Lys Met Ala Leu 325 330 335 Val Ile Asp Asp Arg Leu Lys Val Trp Asp Glu Lys Asp Gln Pro Arg 340 345 350 Val His Val Val Pro Ala Phe Ala Pro Tyr Tyr Ala Pro Gln Ala Glu 355 360 365 Ala Ser Asn Thr Ile Pro Val Leu Cys Val Ala Arg Asn Val Ala Cys 370 375 380 Asn Val Arg Gly Gly Phe Phe Lys Asp Phe Asp Asp Gly Leu Leu Gln 385 390 395 400 Lys Ile Pro Gln Ile Ala Tyr Glu Asp Asp Ile Lys Asp Ile Pro Ser 405 410 415 Pro Pro Asp Val Ser Asn Tyr Leu Val Ser Glu Asp Asp Gly Ser Ile 420 425 430 Ser Asn Gly His Arg Asp Pro Phe Leu Phe Asp Gly Met Ala Asp Ala 435 440 445 Glu Val Glu Arg Lys Leu Lys Asp Ala Leu Ser Ala Ala Ser Thr Ile 450 455 460 Pro Val Thr Thr Ala Asn Leu Asp Pro Arg Leu Thr Ser Leu Gln Tyr 465 470 475 480 Thr Met Val Pro Ser Gly Ser Val Pro Pro Pro Thr Ala Gln Ala Ser 485 490 495 Met Met Pro Phe Pro His Val Gln Phe Pro Gln Pro Ala Thr Leu Val 500 505 510 Lys Pro Met Gly Gln Ala Ala Pro Ser Glu Pro Ser Leu His Ser Ser 515 520 525 Pro Ala Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu Asp Pro Asp 530 535 540 Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp Thr Arg Asp 545 550 555 560 His Ala Ser Ala Glu Pro Pro Phe Pro Val Arg His Pro Val Gln Thr 565 570 575 Ser Ala Pro His Val Pro Ser Ser Arg Gly Val Trp Phe Pro Ala Glu 580 585 590 Glu Glu Ile Gly Ser Gln Pro Leu Asn Arg Val Val Pro Lys Glu Phe 595 600 605 Pro Val Asp Ser Gly Pro Leu Gly Ile Ala Lys Pro Arg Pro His His 610 615 620 Pro Ser Phe Phe Ser Lys Val Glu Ser Ser Ile Ser Ser Asp Arg Ile 625 630 635 640 Leu His Asp Ser His Gln Arg Leu Pro Lys Glu Met Tyr His Arg Asp 645 650 655 Asp Arg Pro Arg Leu Asn His Met Leu Ser Ser Tyr Arg Ser Phe Ser 660 665 670 Gly Asp Asp Ile Pro Phe Ser Arg Ser Phe Ser Ser His Arg Asp Leu 675 680 685 Asp Ser Glu Ser Gly His Ser Val Leu His Ala Asp Thr Pro Val Ala 690 695 700 Val Leu Gln Glu Ile Ala Leu Lys Cys Gly Thr Lys Val Asp Phe Ile 705 710 715 720 Ser Ser Leu Val Ala Ser Thr Glu Leu Gln Phe Ser Met Glu Ala Trp 725 730 735 Phe Ser Gly Lys Lys Ile Gly His Arg Val Gly Arg Thr Arg Lys Glu 740 745 750 Ala Gln Asn Lys Ala Ala Glu Asp Ser Ile Lys His Leu Ala Asp Ile 755 760 765 Tyr Leu Ser Ser Ala Lys Asp Glu Pro Gly Ser Thr Tyr Gly Asp Val 770 775 780 Ser Gly Phe Pro Asn Val Asn Asp Ser Gly Tyr Met Gly Ile Ala Ser 785 790 795 800 Ser Leu Gly Asn Gln Pro Leu Ser Lys Glu Asp Ser Ala Ser Phe Ser 805 810 815 Thr Ala Ser Pro Ser Arg Val Leu Asp Pro Arg Leu Asp Val Ser Lys 820 825 830 Arg Ser Met Gly Ser Ile Ser Ser Leu Lys Glu Leu Cys Met Met Glu 835 840 845 Gly Leu Asp Val Asn Phe Leu Ser Ala Pro Ala Pro Val Ser Thr Asn 850 855 860 Ser Val Gln Lys Asp Glu Val His Ala Gln Val Glu Ile Asp Gly Lys 865 870 875 880 Val Phe Gly Lys Gly Ile Gly Leu Thr Trp Asp Glu Ala Lys Met Gln 885 890 895 Ala Ala Glu Lys Ala Leu Gly Ser Leu Arg Ser Lys Leu Gly Gln Ser 900 905 910 Ile Gln Lys Arg Gln Ser Ser Pro Arg Pro His Gln Gly Phe Ser Asn 915 920 925 Lys Arg Leu Lys Gln Glu Tyr Pro Arg Pro Met Gln Arg Met Pro Ser 930 935 940 Ser Ala Arg Tyr Pro Arg Asn Ala Pro Pro Ile Pro 945 950 955 <210> 13 <211> 960 <212> PRT <213> Glycine max <400> 13 Met Tyr Lys Ser Val Val Tyr Gln Gly Glu Val Val Val Gly Glu Val 1 5 10 15 Asp Val Tyr Pro Glu Glu Asn Asn Asn Asn Asn Asn Lys Asn Tyr Asn 20 25 30 Lys Asn Phe His Val Lys Glu Ile Arg Ile Ser His Phe Ser Gln Pro 35 40 45 Ser Glu Arg Cys Pro Pro Leu Ala Val Leu His Thr Val Thr Ser Cys 50 55 60 Gly Val Cys Phe Lys Met Glu Ser Lys Thr Gln Gln Gln Asp Gly Leu 65 70 75 80 Phe Gln Leu His Ser Leu Cys Ile Arg Glu Asn Lys Thr Ala Val Met 85 90 95 Pro Leu Gly Gly Glu Glu Ile His Leu Val Ala Met His Ser Arg Asn 100 105 110 Asp Asp Arg Pro Cys Phe Trp Gly Phe Ile Val Thr Leu Gly Leu Tyr 115 120 125 Asp Ser Cys Leu Val Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe 130 135 140 Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Met Arg Ser Phe Glu 145 150 155 160 Asp Arg Ile Asp Ala Leu Gln Arg Lys Ile Asn Ser Glu Val Asp Pro 165 170 175 Gln Arg Ile Ser Gly Met Gln Ala Glu Val Lys Arg Tyr Leu Asp Asp 180 185 190 Lys Asn Ile Leu Lys Gln Tyr Ala Glu Asn Asp Gln Val Val Asp Asn 195 200 205 Gly Arg Val Ile Lys Val Gln Ser Glu Ile Val Pro Ala Leu Ser Asp 210 215 220 Ser His Gln Pro Ile Val Arg Pro Leu Ile Arg Leu Gln Asp Lys Asn 225 230 235 240 Ile Ile Leu Thr Arg Ile Asn Pro Gln Ile Arg Asp Thr Ser Val Leu 245 250 255 Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu Thr Ala 260 265 270 Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala Glu Arg 275 280 285 Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser Asn Leu 290 295 300 Ile Asn Ser Lys Glu Leu Leu Gly Arg Ile Val Cys Val Lys Ser Gly 305 310 315 320 Leu Lys Lys Ser Leu Phe Asn Val Phe Gln Asp Gly Ser Cys Asp Pro 325 330 335 Lys Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp Glu Arg 340 345 350 Asp Gln Pro Arg Val His Val Val Pro Ala Phe Ala Pro Tyr Tyr Ala 355 360 365 Pro Gln Ala Glu Ala Ser Asn Thr Ile Pro Val Leu Cys Val Ala Arg 370 375 380 Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Asp Phe Asp Asp 385 390 395 400 Gly Leu Leu Gln Lys Ile Pro Gln Ile Ala Tyr Glu Asp Asp Ile Lys 405 410 415 Asp Val Pro Ser Pro Pro Asp Val Ser Asn Tyr Leu Val Ser Glu Asp 420 425 430 Asp Gly Ser Ile Ser Asn Gly Asn Arg Asp Pro Phe Leu Phe Asp Gly 435 440 445 Met Ala Asp Ala Glu Val Glu Arg Lys Leu Lys Asp Ala Leu Ala Ala 450 455 460 Ala Ser Thr Phe Pro Val Thr Thr Ala Asn Leu Asp Pro Arg Leu Thr 465 470 475 480 Ser Leu Gln Tyr Thr Met Val Pro Ser Gly Ser Val Pro Pro Pro Thr 485 490 495 Ala Gln Ala Ser Met Met Pro Phe Pro His Val Gln Phe Pro Gln Pro 500 505 510 Ala Thr Leu Val Lys Pro Met Gly Gln Ala Ala Pro Ser Asp Pro Ser 515 520 525 Leu His Ser Ser Pro Ala Arg Glu Glu Gly Glu Val Pro Glu Ser Glu 530 535 540 Leu Asp Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln 545 550 555 560 Asp Thr Arg Asp His Ala Ser Ala Glu Pro Pro Phe Pro Val Arg His 565 570 575 Pro Val Gln Ala Ser Ala Pro Arg Val Pro Ser Ser Arg Gly Val Trp 580 585 590 Phe Pro Val Glu Glu Glu Ile Gly Ser Gln Pro Leu Asn Arg Val Val 595 600 605 Pro Lys Glu Phe Pro Val Asp Ser Gly Pro Leu Gly Ile Glu Lys Pro 610 615 620 Arg Leu His His Pro Ser Phe Phe Asn Lys Val Glu Ser Ser Ile Ser 625 630 635 640 Ser Asp Arg Ile Leu His Asp Ser His Gln Arg Leu Pro Lys Glu Met 645 650 655 Tyr His Arg Asp Asp Arg Pro Arg Leu Asn His Met Leu Ser Ser Tyr 660 665 670 Arg Ser Phe Ser Gly Asp Asp Ile Pro Phe Ser Arg Ser Ser Ser Ser 675 680 685 His Arg Asp Leu Asp Ser Glu Ser Gly His Ser Val Leu His Ala Asp 690 695 700 Thr Pro Val Ala Val Leu His Glu Ile Ala Leu Lys Cys Gly Thr Lys 705 710 715 720 Val Asp Phe Met Ser Ser Leu Val Ala Ser Thr Glu Leu Lys Phe Ser 725 730 735 Leu Glu Ala Trp Phe Ser Gly Lys Lys Ile Gly His Gly Phe Gly Arg 740 745 750 Thr Arg Lys Glu Ala Gln Asn Lys Ala Ala Lys Asp Ser Ile Glu His 755 760 765 Leu Ala Asp Ile Tyr Leu Ser Ser Ala Lys Asp Glu Pro Gly Ser Thr 770 775 780 Tyr Gly Asp Val Ser Gly Phe Pro Asn Val Asn Asp Asn Gly Tyr Met 785 790 795 800 Gly Ile Ala Ser Ser Leu Gly Asn Gln Pro Leu Ser Lys Glu Asp Ser 805 810 815 Ala Ser Phe Ser Ser Ala Ser Pro Ser Arg Ala Leu Asp Pro Arg Leu 820 825 830 Asp Val Ser Lys Arg Ser Met Gly Ser Ile Ser Ala Leu Lys Glu Leu 835 840 845 Cys Met Met Glu Gly Leu Gly Val Asn Phe Leu Ser Thr Pro Ala Pro 850 855 860 Val Ser Thr Asn Ser Val Gln Lys Asp Glu Val His Ala Gln Val Glu 865 870 875 880 Ile Asp Gly Lys Ile Phe Gly Lys Gly Ile Gly Leu Thr Trp Asp Glu 885 890 895 Ala Lys Met Gln Ala Ala Glu Lys Ala Leu Gly Asn Leu Arg Ser Lys 900 905 910 Leu Gly Gln Ser Ile Gln Lys Met Gln Ser Ser Pro Arg Pro His Gln 915 920 925 Gly Phe Ser Asn Lys Arg Leu Lys Gln Glu Tyr Pro Arg Thr Met Gln 930 935 940 Arg Met Pro Ser Ser Ala Arg Tyr Pro Arg Asn Ala Pro Pro Ile Pro 945 950 955 960 <210> 14 <211> 960 <212> PRT <213> Glycine max <400> 14 Met Pro Thr Ser Met Val Tyr His Gly Glu Met Ala Val Gly Glu Val 1 5 10 15 Lys Ile Tyr Pro Glu Glu Asn Lys Asn Met Asp Leu Lys Glu Ile Arg 20 25 30 Ile Ser His Phe Ser Gln Pro Ser Glu Arg Cys Pro Pro Leu Ala Val 35 40 45 Leu His Thr Ile Thr Ser Phe Gly Ile Cys Phe Lys Met Glu Ser Ser 50 55 60 Thr Ser Gln Lys Arg Gln Gln Gln Asp Ala Leu Phe His Leu His Ser 65 70 75 80 Ser Cys Ile Arg Glu Asn Lys Thr Ala Val Met Pro Val Arg Gly Glu 85 90 95 Glu Ile His Leu Val Ala Met Tyr Ser Arg Asn Asn Asp Arg Pro Cys 100 105 110 Phe Trp Gly Phe Ile Val Ala Ser Gly Leu Tyr Asn Ser Cys Leu Thr 115 120 125 Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe Asp Leu Asp Glu Thr 130 135 140 Leu Val Val Ala Asn Thr Met Arg Ser Phe Glu Asp Lys Ile Glu Val 145 150 155 160 Leu His Arg Lys Met Asn Ser Glu Val Asn Pro Gln Gln Ile Ser Ala 165 170 175 Met Gln Ala Glu Ile Lys Arg Tyr Leu Asp Asp Lys Asn Ile Leu Lys 180 185 190 Glu Tyr Ala Glu Asn Asp Gln Val Val Asp Asn Gly Lys Val Ile Lys 195 200 205 Ile Gln Ser Glu Ser Val Pro Ala Leu Ser Asp Ser His Gln Pro Ile 210 215 220 Val Arg Pro Leu Ile Arg Leu Gln Glu Lys Asn Ile Ile Leu Thr Arg 225 230 235 240 Ile Asn Pro Gln Ile Arg Asp Thr Ser Val Leu Val Arg Leu Arg Pro 245 250 255 Ala Trp Glu Asp Leu Arg Ser Tyr Leu Thr Ala Arg Gly Arg Lys Arg 260 265 270 Phe Glu Val Phe Val Cys Thr Met Ala Glu Arg Asp Tyr Ala Leu Glu 275 280 285 Met Trp Arg Leu Leu Asp Pro Glu Leu Asn Leu Ile Asn Ser Lys Glu 290 295 300 Leu Leu Asp Arg Ile Val Cys Val Lys Ser Gly Leu Lys Lys Ser Leu 305 310 315 320 Phe Asn Val Phe Gln Asn Gly Leu Cys His Leu Lys Met Ala Leu Val 325 330 335 Ile Asp Asp Arg Leu Lys Val Trp Asp Glu Lys Asp Gln Pro Arg Val 340 345 350 His Val Val Pro Ala Phe Ala Pro Tyr Tyr Thr Pro Gln Ala Glu Ala 355 360 365 Ser Asn Ala Val Pro Phe Leu Cys Leu Ala Arg Asn Val Ala Cys Asn 370 375 380 Val Arg Gly Gly Phe Phe Lys Asp Phe Asp Asp Gly Leu Leu Gln Lys 385 390 395 400 Ile Pro Leu Ile Ala Tyr Glu Asp Asp Ile Lys Asp Ile Pro Ser Pro 405 410 415 Asp Val Ser Asn Tyr Leu Val Ser Glu Asp Asp Ala Ser Ala Ser Asn 420 425 430 Gly Asn Lys Asn Leu Leu Leu Phe Asp Gly Met Ala Asp Ala Glu Val 435 440 445 Glu Arg Arg Leu Lys Asp Ala Ile Ser Ala Ser Ser Thr Ile Leu Ala 450 455 460 Leu Thr Ala Asn Ile Asp Pro Arg Leu Ala Phe Thr Ser Ser Leu Gln 465 470 475 480 Tyr Thr Met Val Ser Ser Ser Gly Thr Val Pro Pro Pro Thr Ala Gln 485 490 495 Ala Ser Val Val Gln Phe Gly Asn Val Gln Phe Pro Gln Pro Asn Thr 500 505 510 Leu Val Lys Pro Met Ser Gln Val Thr His Pro Gly Leu Ser Leu His 515 520 525 Ser Ser Pro Ala Arg Glu Glu Gly Glu Leu Pro Glu Ser Glu Leu Asp 530 535 540 Leu Asp Thr Arg Arg Arg Phe Leu Ile Leu Gln His Gly Gln Asp Thr 545 550 555 560 Arg Glu Arg Met Ala Ser Glu Pro Pro Phe Pro Val Arg His Pro Ala 565 570 575 Gln Val Ser Ala Pro Ala Ser Ser Val Pro Ser Arg Arg Gly Trp Phe 580 585 590 Ser Val Glu Glu Glu Met Gly Pro Gln Gln Leu Asn Leu Pro Val Pro 595 600 605 Lys Glu Phe Pro Val Asp Ser Glu Pro Phe His Ile Glu Lys Arg Trp 610 615 620 Pro Arg His Pro Ser Phe Phe Ser Lys Val Gly Asp Ser Ile Ser Ser 625 630 635 640 Asp Arg Val Phe His Glu Ser His Gln Arg Leu Pro Lys Glu Val His 645 650 655 His Arg Asp Asp Arg Ser Arg Leu Ser Gln Ser Leu Ser Ser Tyr His 660 665 670 Ser Leu Pro Gly Asp Asp Ile Pro Leu Ser Gly Ser Ser Tyr Ser Asn 675 680 685 Arg Asp Phe Asp Ser Glu Ser Gly Arg Ser Leu Phe His Ala Asp Thr 690 695 700 Thr Ala Gly Val Leu Gln Glu Ile Ala Leu Asn Cys Gly Thr Lys Val 705 710 715 720 Glu Phe Leu Ser Ser Leu Val Ala Ser Thr Glu Leu Gln Phe Ser Ile 725 730 735 Glu Ala Trp Phe Ala Gly Lys Lys Ile Gly Glu Gly Phe Gly Arg Thr 740 745 750 Arg Arg Glu Ala Gln Ser Lys Ala Ala Gly Cys Ser Ile Lys Gln Leu 755 760 765 Ala Asp Ile Tyr Met Ser His Ala Lys Asp Asp Ser Gly Ser Thr Tyr 770 775 780 Gly Asp Val Ser Gly Phe His Gly Ser Asn Asn Asp Gly Phe Val Ser 785 790 795 800 Ser Gly Asn Ser Leu Gly Asn Gln Leu Leu Pro Lys Glu Glu Ser Gly 805 810 815 Ser Phe Ser Thr Ala Ser Glu Ser Ser Arg Val Ser Asp Ser Arg Leu 820 825 830 Glu Val Ser Lys Arg Ser Thr Asp Ser Ile Ser Ala Leu Lys Glu Leu 835 840 845 Cys Met Met Glu Gly Leu Ala Ala Ser Phe Gln Ser Pro Pro Ala Ser 850 855 860 Ala Ser Thr His Leu Thr Gln Lys Asp Glu Val His Ala Gln Val Glu 865 870 875 880 Ile Asp Gly Gln Ile Phe Gly Lys Gly Phe Gly Val Thr Trp Glu Glu 885 890 895 Ala Lys Met Gln Ala Ala Lys Lys Ala Leu Gly Ser Leu Arg Thr Met 900 905 910 Phe Asn Gln Gly Ser Leu Lys Arg His Gly Ser Pro Arg Ser Met Gln 915 920 925 Gly Leu Ala Asn Lys Arg Leu Lys Pro Glu Tyr Pro Pro Thr Leu Gln 930 935 940 Arg Val Pro Tyr Ser Ala Arg Tyr Pro Arg Asn Ala Pro Leu Val Pro 945 950 955 960 <210> 15 <211> 958 <212> PRT <213> Glycine max <400> 15 Met Lys Arg Ser Met Val Tyr His Gly Glu Met Glu Val Gly Glu Val 1 5 10 15 Glu Ile Tyr Pro Glu Glu Lys Lys Asn Ile Asp Leu Lys Glu Ile Arg 20 25 30 Ile Ser His Phe Ser Gln Pro Ser Glu Arg Cys Pro Pro Leu Ala Val 35 40 45 Leu His Thr Ile Thr Ser Phe Gly Ile Cys Phe Lys Met Glu Ser Ser 50 55 60 Thr Ser Gln Thr Arg Gln Gln Gln Asp Val Leu Phe His Leu His Ser 65 70 75 80 Ser Cys Ile Arg Glu Asn Lys Thr Ala Val Met Pro Leu Arg Gly Glu 85 90 95 Glu Ile His Leu Val Ala Met Tyr Ser Arg Asn Asn Asp Arg Pro Cys 100 105 110 Phe Trp Gly Phe Ile Val Ala Ser Gly Leu Tyr Asn Ser Cys Leu Thr 115 120 125 Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe Asp Leu Asp Glu Thr 130 135 140 Leu Val Val Ala Asn Thr Met Arg Ser Phe Glu Asp Lys Ile Glu Val 145 150 155 160 Leu His Arg Lys Met Asn Ser Glu Val Asn Pro Gln Arg Ile Ser Thr 165 170 175 Met Gln Ala Glu Ile Lys Arg Tyr Leu Asp Asp Lys Asn Ile Leu Lys 180 185 190 Glu Tyr Ala Glu Asn Asp Gln Val Val Asp Asn Gly Lys Val Ile Lys 195 200 205 Ile Gln Ser Glu Ile Val Pro Ala Leu Ser Asp Ser His Gln Pro Ile 210 215 220 Val Arg Pro Leu Ile Arg Leu Gln Glu Lys Asn Ile Ile Leu Thr Arg 225 230 235 240 Ile Asn Pro Gln Ile Arg Asp Thr Ser Val Leu Val Arg Leu Arg Pro 245 250 255 Ala Trp Glu Asp Leu Arg Ser Tyr Leu Thr Ala Arg Gly Arg Lys Arg 260 265 270 Phe Glu Val Phe Val Cys Thr Met Ala Glu Arg Asp Tyr Ala Leu Glu 275 280 285 Met Trp Arg Leu Leu Asp Pro Glu Leu Asn Leu Ile Asn Ser Lys Glu 290 295 300 Leu Leu Asp Arg Ile Val Cys Val Lys Ser Gly Leu Lys Lys Ser Leu 305 310 315 320 Phe Asn Val Phe Gln Asn Gly Leu Cys His Leu Lys Met Ala Leu Val 325 330 335 Ile Asp Asp Arg Leu Lys Val Trp Asp Glu Lys Asp Gln Pro Gln Val 340 345 350 His Val Val Pro Ala Phe Ala Pro Tyr Tyr Ala Pro Gln Ala Glu Ala 355 360 365 Ser Asn Ala Val Pro Thr Leu Cys Leu Ala Arg Ser Val Ala Cys Asn 370 375 380 Val Arg Gly Gly Phe Phe Lys Asp Phe Asp Asp Gly Leu Leu Gln Lys 385 390 395 400 Ile Pro Leu Ile Ala Tyr Glu Asp Asp Ile Lys Asp Ile Pro Ser Pro 405 410 415 Pro Asp Val Ser Asn Tyr Leu Val Ser Glu Asp Asp Ala Ser Ala Ser 420 425 430 Asn Gly Asn Lys Asn Leu Leu Leu Phe Asp Gly Met Ala Asp Ala Glu 435 440 445 Val Glu Arg Arg Leu Lys Asp Ala Ile Ser Ala Ser Ser Thr Val Pro 450 455 460 Ala Met Thr Thr Asn Leu Asp Pro Arg Leu Ala Phe Asn Ser Ser Leu 465 470 475 480 Gln Tyr Thr Met Val Ser Ser Ser Gly Thr Val Pro Pro Pro Thr Ala 485 490 495 Gln Ala Ser Ile Val Gln Phe Gly Asn Val Gln Phe Pro Gln Pro Asn 500 505 510 Thr Leu Val Lys Pro Ile Cys Gln Val Thr Pro Pro Gly Pro Ser Leu 515 520 525 His Ser Ser Pro Ala Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu 530 535 540 Asp Leu Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp 545 550 555 560 Thr Arg Glu His Thr Ser Ser Glu Pro Pro Leu Pro Val Arg His Pro 565 570 575 Thr Gln Val Ser Ala Pro Ser Val Pro Ser Arg Arg Gly Trp Phe Ser 580 585 590 Val Glu Glu Glu Met Gly Pro Gln Gln Leu Asn Gln Leu Val Pro Lys 595 600 605 Glu Phe Pro Val Gly Ser Glu Pro Leu His Ile Glu Lys Arg Trp Pro 610 615 620 Arg His Pro Ser Leu Phe Ser Lys Val Asp Asp Ser Val Ser Ser Asp 625 630 635 640 Arg Val Phe His Glu Ser His Gln Arg Leu Pro Lys Glu Val His His 645 650 655 Arg Asp Asp His Ser Arg Leu Ser Gln Ser Leu Ser Ser Tyr His Ser 660 665 670 Phe Pro Gly Asp Asp Ile Pro Leu Ser Gly Ser Ser Tyr Ser Asn Arg 675 680 685 Asp Phe Asp Ser Glu Ser Gly Arg Ser Leu Phe His Ala Asp Ile Thr 690 695 700 Ala Gly Val Leu Gln Glu Ile Ala Leu Lys Cys Gly Thr Lys Val Glu 705 710 715 720 Phe Leu Ser Ser Leu Val Ala Ser Thr Ala Leu Gln Phe Ser Ile Glu 725 730 735 Ala Trp Phe Ala Gly Lys Lys Val Gly Glu Gly Phe Gly Arg Thr Arg 740 745 750 Arg Glu Ala Gln Asn Lys Ala Ala Glu Cys Ser Ile Lys Gln Leu Ala 755 760 765 Asp Ile Tyr Met Ser His Ala Lys Asp Asp Ser Gly Ser Thr Tyr Gly 770 775 780 Asp Val Ser Gly Phe His Gly Ser Asn Asn Asn Gly Phe Val Ser Ser 785 790 795 800 Gly Asn Ser Leu Gly Asn Gln Leu Leu Pro Lys Glu Ser Val Ser Phe 805 810 815 Ser Thr Ser Ser Asp Ser Ser Arg Val Ser Asp Pro Arg Leu Glu Val 820 825 830 Ser Lys Arg Ser Thr Asp Ser Ile Ser Ala Leu Lys Glu Phe Cys Met 835 840 845 Met Glu Gly Leu Ala Ala Asn Phe Gln Ser Ser Pro Ala Pro Ala Ser 850 855 860 Thr His Phe Ala Gln Lys Asp Glu Val His Ala Gln Val Glu Ile Asp 865 870 875 880 Gly Gln Ile Phe Gly Lys Gly Phe Gly Leu Thr Trp Glu Glu Ala Lys 885 890 895 Met Gln Ala Ala Lys Lys Ala Leu Glu Ser Leu Arg Thr Met Phe Asn 900 905 910 Gln Gly Thr Arg Lys Arg His Gly Ser Pro Arg Ser Met Gln Gly Leu 915 920 925 Ala Asn Lys Arg Leu Lys Gln Glu Tyr Pro Arg Thr Leu Gln Arg Ile 930 935 940 Pro Tyr Ser Ala Arg Tyr Pro Arg Asn Ala Pro Leu Val Pro 945 950 955 <210> 16 <211> 1023 <212> PRT <213> Beta vulgaris <400> 16 Met Ile Lys Ser Val Val Tyr Glu Gly Glu Asn Leu Leu Gly Glu Val 1 5 10 15 Glu Ile Tyr Phe Gln Asn Asn Asn Asn Asn Lys Asn Leu Glu Leu Met 20 25 30 Lys Gly Met Arg Ile Ser His Tyr Ser Glu Met Ser Glu Arg Cys Pro 35 40 45 Pro Leu Ala Val Leu His Thr Ile Thr Lys Ser Ser Gly Gly Ile Cys 50 55 60 Phe Lys Met Met Glu Ser Ser Ser His Thr Ser Ser Asn Asn Asn Asn 65 70 75 80 Asn Asn Lys Phe Tyr Phe Gln Gln Gln Gln Gln Glu Ser Gln Leu Leu 85 90 95 Ala Met His Ser Asn Cys Ile Arg Asp Asn Lys Thr Ala Val Val Pro 100 105 110 Leu Gly Glu Gln Glu Ile His Leu Val Ala Leu Arg Ser Arg Arg Met 115 120 125 Ala Gly Val Thr Pro Cys Phe Trp Gly Phe Ser Val Ala Pro Gly Leu 130 135 140 Tyr Glu Ser Cys Leu Gly Leu Leu Asn Leu Arg Cys Leu Gly Ile Val 145 150 155 160 Phe Asp Leu Asp Glu Thr Leu Ile Val Ala Asn Thr Leu Arg Ser Phe 165 170 175 Glu Asp Arg Ile Glu Ala Leu Gln Arg Lys Ile Ser Val Glu Ala Asp 180 185 190 Pro Gln Arg Ile Ala Gly Met Val Ala Glu Val Lys Arg Tyr Gln Glu 195 200 205 Asp Lys Ser Ile Leu Lys Gln Tyr Ala Glu Thr Asp Gln Val Val Asp 210 215 220 Asn Gly Lys Val His Lys Ile Gln Ala Glu Val Ile Pro Ala Leu Ser 225 230 235 240 Asp Asn His Gln Thr Val Ile Arg Pro Leu Ile Arg Leu Gln Asp Lys 245 250 255 Asn Ile Val Leu Thr Arg Ile Asn Pro Gln Ile Arg Asp Thr Ser Val 260 265 270 Leu Val Arg Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu Thr 275 280 285 Ala Arg Gly Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala Glu 290 295 300 Arg Asp Tyr Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser Asn 305 310 315 320 Leu Ile Cys Ala Arg Glu Leu Leu Asp Arg Ile Val Cys Val Lys Ser 325 330 335 Gly Ser Lys Lys Ser Leu Phe Asn Val Phe His Gly Gly Ile Cys His 340 345 350 Pro Lys Met Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp Glu 355 360 365 Lys Asp Gln Pro Arg Val His Val Val Pro Ala Phe Ala Pro Tyr Tyr 370 375 380 Ala Pro Gln Ala Glu Ala Asn Asn Ala Ile Pro Val Leu Cys Val Ala 385 390 395 400 Arg Asn Val Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Glu Phe Asp 405 410 415 Glu Gly Leu Leu Gln Arg Val Ser Glu Val Ser Phe Glu Asp Asp Pro 420 425 430 Arg Asp Ile Pro Ser Pro Pro Asp Val Ser Asn Tyr Leu Val Ser Glu 435 440 445 Asp Asp Gly Ser Gly Ser Asn Gly Ile Lys Glu Ser Met Thr Phe Asp 450 455 460 Gly Met Ala Asp Ala Glu Val Glu Arg Arg Leu Lys Glu Ala Val Leu 465 470 475 480 Ser Ser Ser Ser Ala Ser Pro Leu Pro Ser Ala Asn Thr Pro Ala Thr 485 490 495 Val Asn Phe Asp His Arg Leu Ala Ser Ser Leu Pro Phe Ala Val Ala 500 505 510 Thr Ser Ala Leu Ala Ile Pro Gln Pro Ala Pro Gln Ala Thr Ile Thr 515 520 525 Pro Tyr His Asn Asn Leu Phe Ser Gln Ala Gly Pro Leu Ala Arg Pro 530 535 540 Leu Gly Asn Ile Gly Pro Gln Asp Ile Gly Leu His Asn Ser Pro Ala 545 550 555 560 Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu Asp Pro Asp Thr Arg 565 570 575 Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp Met Arg Glu Gly Pro 580 585 590 Pro Asn Glu Pro Pro Phe Pro Ala Arg Thr Pro Val Gln Ala Pro Val 595 600 605 Thr Gly Pro Val Pro Val Ser Val Pro Gly Pro Val Pro Val Pro Gly 610 615 620 Pro Gly Pro Val Ser Val Pro Val Pro Gly Pro Ile Pro Ser Pro Val 625 630 635 640 Pro Val Pro Val Ser Gly Thr Val Pro Gly Pro Gly Pro Arg Val Gln 645 650 655 Ser Arg Gly Ser Trp Phe Pro Val Glu Asp His Ile Ser Gln Gly Pro 660 665 670 Leu Ser Arg Val Ala Ala Lys Glu Phe Pro Val Ala Pro Asp Ala Ser 675 680 685 Pro Val Glu Lys Gln Arg Pro Pro Pro Pro Ser Phe Pro Arg Lys Val 690 695 700 Glu Ser Leu Gly Trp Ser Asp Arg Asn Tyr Ala Glu Lys Gln Arg Leu 705 710 715 720 Pro Arg Glu Ala Leu Arg Arg Asp Asp Arg Leu Arg Ser Asn Tyr Ser 725 730 735 Leu Pro Ser His Gln Ser Phe Arg Gly Asp Glu Ile Ser Leu Ser Arg 740 745 750 Ser Ala Ser Ser Asn Lys Asp Phe Glu Val Glu Pro Glu Arg Gly Ser 755 760 765 Ser Phe Ala Glu Ser Pro Ser Ile Ala Leu His Asp Ile Ala Met Lys 770 775 780 Cys Gly Thr Lys Val Glu Phe Lys Thr Gly Leu Val Ala Thr Pro Glu 785 790 795 800 Leu Lys Phe Leu Leu Glu Ala Tyr Phe Ala Gly Asp Lys Ile Gly Glu 805 810 815 Gly Thr Gly Thr Thr Arg Arg Glu Ala Gln His Arg Ala Ala Glu Ala 820 825 830 Ala Leu Met Asn Leu Ala Asp Lys Tyr Leu Thr His Ile Lys Ser Asp 835 840 845 Ser Ser Thr Pro Gln Ser Asp Thr Ser Arg Gly His Ser Pro Ile Asp 850 855 860 Thr Gly Phe Val Ser Asp Ala Asn Ser His Gly Asp Gly Ile Ser Arg 865 870 875 880 Lys Glu Asp Ile Ile Pro Ser Ser Glu Met Thr Gly Leu Asp Asp Ser 885 890 895 Asn Val Asp Gly Ser Lys Asn Ser Met Gly Ser Val Ser Val Leu Lys 900 905 910 Glu Leu Cys Leu Arg Glu Gly Leu Gly Val Asp Phe Lys Gly Gln Ser 915 920 925 Pro Thr Ser Thr Asn Ser Val Asp Arg Asp Glu Ile His Ala Glu Val 930 935 940 Glu Ile Asn Gly Gln Val Leu Gly Lys Gly Thr Gly Leu Thr Trp Asp 945 950 955 960 Glu Ala Lys Met Gln Ala Ala Glu Met Ala Leu Thr Ser Leu Asn Ser 965 970 975 Methionine Isoleucine Glycine Glutamine Phenylalanine Asparagine Lysine Arginine Proline Serine Serine Proline Arginine Leucine Leucine Glutamine 980 985 990 Glycine Methionine Proline Asparagine Lysine Arginine Leucine Lysine Proline Glutamic acid Tyrosine Proline Arginine Valine Valine Aspartic acid 995 1000 1005 Histidine Leucine Proline Serine Serine Arginine Tyrosine Proline Arginine Asparagine Alanine Serine Proline Valine Proline 1010 1015 1020 <210> 17 <211> 881 <212> PRT <213> Solanum tuberosum <400> 17 Methionine Phenylalanine Lysine Serine Threonine Valine Valine Leucine Tyrosine Glutamic acid Glycine Glutamic acid Arginine Leucine Valine Glycine 1 5 10 15 Glutamic acid Valine Glutamic acid Isoleucine Tyrosine Cysteine Glutamic acid Lysine Glycine Valine Leucine Tryptophan Glycine Glutamic acid Lysine Valine 20 25 30 Isoleucine Arginine Isoleucine Serine Histidine Tyrosine Serine Proline Serine Serine Glutamic acid Arginine Cysteine Proline Proline Leucine 35 40 45 Alanine Valine Leucine Histidine Threonine Valine Threonine Threonine Glycine Leucine Serine Phenylalanine Lysine Leucine Glutamic acid Proline 50 55 60 Threonine Lysine Serine Lysine Proline Leucine Threonine Glutamine Aspartic acid Serine Proline Leucine Threonine Leucine Leucine Histidine 65 70 75 80 Serine Threonine Cysteine Leucine Arginine Aspartic acid Asparagine Lysine Threonine Alanine Valine Methionine Serine Leucine Glycine Arginine 85 90 95 Glu Glu Leu His Leu Val Ala Met Gln Ser Lys Asn Ile Gly Gly Gln 100 105 110 Cys Pro Cys Phe Trp Gly Phe Lys Val Ala Ser Gly Leu Tyr Asp Ser 115 120 125 Cys Leu Thr Met Leu Asn Leu Arg Cys Leu Gly Ile Val Phe Asp Leu 130 135 140 Asp Glu Thr Leu Ile Val Ala Asn Thr Met Arg Ser Phe Glu Asp Arg 145 150 155 160 Ile Glu Ala Leu Gln Arg Lys Ile Asn Ser Glu Ser Asp Pro Gln Arg 165 170 175 Ala Ser Val Met Leu Ala Glu Val Lys Arg Tyr Gln Glu Asp Lys Ile 180 185 190 Ile Leu Lys Gln Tyr Ala Glu Asn Asp Gln Val Val Asp Asn Gly Lys 195 200 205 Val Ile Lys Ser Gln Ser Glu Val Phe Pro Ala Leu Ser Asp Asn His 210 215 220 Gln Pro Ile Val Arg Pro Leu Ile Arg Leu Gln Asp Arg Asn Ile Ile 225 230 235 240 Leu Thr Arg Ile Asn Pro Met Ile Arg Asp Thr Ser Val Leu Val Arg 245 250 255 Leu Arg Pro Ala Trp Glu Asp Leu Arg Ser Tyr Leu Thr Ala Arg Gly 260 265 270 Arg Lys Arg Phe Glu Val Tyr Val Cys Thr Met Ala Glu Arg Asp Tyr 275 280 285 Ala Leu Glu Met Trp Arg Leu Leu Asp Pro Asp Ser Asn Leu Ile Asn 290 295 300 Ser Gln Glu Leu Leu Asp Arg Ile Val Cys Val Lys Ser Gly Leu Arg 305 310 315 320 Lys Ser Leu Phe Asn Val Phe Gln Asp Gly Asn Cys His Pro Lys Met 325 330 335 Ala Leu Val Ile Asp Asp Arg Leu Lys Val Trp Asp Asp Lys Asp Gln 340 345 350 Pro Arg Val His Val Val Pro Ala Phe Ala Pro Tyr Phe Ala Pro Gln 355 360 365 Ala Glu Gly Asn Asn Ser Val Pro Val Leu Cys Val Ala Arg Asn Val 370 375 380 Ala Cys Asn Val Arg Gly Gly Phe Phe Lys Asp Phe Asp Glu Gly Leu 385 390 395 400 Leu Gln Arg Ile Ser Glu Val Ala Tyr Glu Asp Asp Ile Lys Gln Val 405 410 415 Pro Ser Ala Pro Asp Val Ser Asn Tyr Leu Ile Ser Glu Asp Asp Pro 420 425 430 Ser Ala Val Asn Gly Asn Lys Asp Ser Leu Gly Phe Asp Gly Met Ala 435 440 445 Asp Ser Glu Val Glu Arg Arg Leu Lys Glu Ala Met Leu Ala Ser Thr 450 455 460 Ser Val Pro Ser Gln Met Thr Asn Leu Asp Pro Arg Leu Val Pro Ala 465 470 475 480 Leu Gln Tyr Pro Val Pro Pro Val Ile Ser Gln Pro Ser Ile Gln Ser 485 490 495 Pro Val Val Pro Phe Pro Thr Gln His Leu Pro Gln Val Thr Ser Val 500 505 510 Leu Lys Ser Ser Val Thr Gln Ile Ser Pro Gln Asp Thr Ser Leu Gln 515 520 525 Ser Ser Pro Ala Arg Glu Glu Gly Glu Val Pro Glu Ser Glu Leu Asp 530 535 540 Pro Asp Thr Arg Arg Arg Leu Leu Ile Leu Gln His Gly Gln Asp Thr 545 550 555 560 Arg Asp Gln Val Ser Ser Glu Pro Lys Phe Pro Met Gly Thr Pro Leu 565 570 575 Gln Val Ser Val Pro Pro Arg Val Gln Pro His Gly Trp Phe Pro Ala 580 585 590 Glu Glu Glu Met Ser Pro Arg Gln Leu Asn Arg Pro Leu Pro Pro Lys 595 600 605 Glu Phe Pro Leu Asn Pro Glu Ser Met His Ile Asn Lys His Arg Pro 610 615 620 Pro His Pro Pro Phe Leu Pro Lys Met Glu Thr Ser Met Pro Ser Asp 625 630 635 640 Arg Val Leu Phe Glu Asn Gln Arg Leu Pro Lys Glu Val Ile Pro Arg 645 650 655 Asp Asp Arg Met Arg Phe Ser Gln Ser Gln Pro Ser Phe Arg Pro Pro 660 665 670 Gly Glu Glu Val Pro Leu Gly Arg Ser Ser Ser Ser Asn Arg Val Leu 675 680 685 Asp Leu Glu Pro Gly His Tyr Asp Pro Tyr Leu Glu Thr Pro Ala Gly 690 695 700 Ala Leu Gln Asp Ile Ala Phe Lys Cys Gly Ala Lys Val Glu Phe Arg 705 710 715 720 Ser Ser Phe Leu Ser Ser Pro Glu Leu Gln Phe Ser Leu Glu Val Leu 725 730 735 Phe Ala Gly Glu Lys Val Gly Glu Gly Thr Gly Arg Thr Arg Arg Glu 740 745 750 Ala Gln Arg Arg Ala Ala Glu Glu Ser Leu Met Tyr Leu Ala Asp Lys 755 760 765 Tyr Leu Ser Cys Ile Lys Pro Asp Ser Ser Ser Thr Gln Gly Asp Gly 770 775 780 Phe Arg Phe Pro Asn Ala Ser Asp Asn Gly Phe Val Asp Asn Met Ser 785 790 795 800 Pro Phe Gly Tyr Gln Asp Arg Val Ser His Ser Phe Ala Ser Glu Pro 805 810 815 Pro Arg Val Leu Asp Pro Arg Leu Glu Val Phe Lys Lys Ser Val Gly 820 825 830 Ser Val Gly Ala Leu Arg Glu Leu Cys Ala Ile Glu Gly Leu Gly Leu 835 840 845 Ala Phe Gln Thr Gln Pro Gln Leu Ser Ala Asn Pro Gly Gln Lys Ser 850 855 860 Glu Ile Tyr Ala Gln Val Ala His Leu Leu Pro Phe Leu Leu Cys Tyr 865 870 875 880 Cys <210> 18 <211> 2904 <212> DNA <213> Artificial Sequence <220> <223> CDS of Arabidopsis_thaliana_AtCPL1 <400> 18 atgtatagta ataatagagt agaagtgttt catggtgatg gaagacttgg agaattggag 60 atataccctt caagggaatt gaatcagcaa caagatgatg tgatgaagca gaggaagaag 120 aaacagaggg aagtaatgga gctagccaag atgggaatca gaataagcca cttttctcaa 180 tctggcgaga ggtgtcctcc tcttgcaata cttactacaa tttcatcttg tggcctttgt 240 ttcaaattag aagcttcacc ttctccagct caggagtcac tcagtttatt ctactcgtcc 300 tgtctcaggg acaacaagac agcagtaatg ctcttgggtg gagaagagct ccatttggtt 360 gctatgtact cggaaaatat caagaatgac cgtccttgtt tctgggcatt tagtgttgct 420 cctggaattt acgattcctg tcttgttatg ttgaatctta gatgtctggg tattgtcttt 480 gatcttgatg aaacccttgt agtggcgaat accatgcgct catttgagga taagattgac 540 gggtttcagc ggagaataaa caacgagatg gaccctcaac gccttgccgt tatagtggct 600 gagatgaagc gttatcaaga tgacaaaaat ctattgaagc aatatattga aagtgaccag 660 gttgttgaaa acggggaggt gataaaggtg caatctgaaa ttgttcctgc cttgtctgac 720 aaccatcagc ctcttgttcg ccctctgata aggttgcaag agaagaatat tattctgact 780 cgcattaacc caatgattcg tgatacaagt gttcttgtga gaatgaggcc ctcatgggag 840 gaacttcgaa gctatttgac agcaaaaggg cgtaagcgtt ttgaagtata tgtttgcacg 900 atggctgaaa gagattacgc cttagagatg tggaggctcc ttgatccaga agggaatttg 960 ataaacacaa atgacttgct tgctcgcatc gtttgtgtga aatctggttt taaaaagtca 1020 ctgttcaatg tgtttctcga tggaacctgc catccaaaga tggcattggt aattgatgat 1080 cgattgaaag tttgggatga gaaggatcag ccgagggtac atgtggttcc tgcattcgct 1140 ccctattatt ctcctcaagc tgaagctgct gcaacaccag tactatgtgt tgccaggaac 1200 gttgcctgtg gtgtcagagg tggatttttc agggattttg atgatagtct gctaccaagg 1260 attgctgaaa tttcttatga gaatgatgct gaggatattc cttctccgcc tgatgtcagc 1320 cattatttgg tgtcggagga tgatacatcg ggtttaaatg ggaacaaaga tccactttcc 1380 tttgacggga tggctgatac tgaagtggag agaagactga aggaggcaat ttctgcatct 1440 tcagctgtcc ttccggcggc aaatatagat ccgaggatag ctgctcccgt tcagttcccc 1500 atggcttctg cttcttctgt ttcagttcca gtaccagtac aagtcgtgca acaagcaata 1560 caaccttcag ctatggcctt tccaagtatt ccatttcaac aacctcaaca accgacatca 1620 atagctaaac acttggttcc ttcagaacca agcttgcaga gttctcctgc tagagaggaa 1680 ggtgaggtac ctgaatcaga attagatcca gatactagga ggagactcct catattgcag 1740 catggacaag atactaggga tcctgctcca agtgaacctt catttcctca gagacctcca 1800 gttcaagctc caccctcaca tgtgcaatca agaaatggct ggtttcctgt tgaggaggag 1860 atggatcctg ctcaaattcg tcgagcagtc tcaaaagaat atccgttgga ttctgaaatg 1920 attcatatgg aaaagcacag gcctcgtcat ccatcatttt tttctaagat tgataactca 1980 actcagtctg acaggatgct tcatgagaat cgcaggccgc caaaggagtc tctccggaga 2040 gatgaacagt tacgttcaaa taacaatcta cctgactctc atcctttcta tggggaggat 2100 gcgtcttgga atcaatcttc ctctaggaac agtgatcttg acttcctacc tgaacgaagt 2160 gtctcagcaa cggagacttc agctgatgtt ctacacggaa ttgctatcaa atgtggagct 2220 aaggtggagt acaaaccaag tttagtttct agtacagatt tgcggttctc tgttgaggct 2280 tggctttcta atcaaaaaat tggagaaggg attggcaaat cgagaagaga agccctgcat 2340 aaggctgctg aagcttctat acagaattta gctgatggat atatgcgtgc aaatggtgac 2400 ccagggccca gccacagaga tgctaccccc ttcaccaatg aaaatataag tatgggaaac 2460 gcaaatgcgc ttaataatca gccatttgct agagatgaaa cagcgttgcc agtttcttct 2520 agacctacag atccgagatt agaaggttct atgaggcaca ctggctccat tactgcactc 2580 agggaattgt gtgcatcgga gggtcttgag atggcttttc aatctcagcg tcagcttcca 2640 tctgacatgg tccacagaga tgaattacat gctcaggttg aaatagatgg gcgtgttgta 2700 ggggaaggag ttggatcgac atgggacgaa gctagaatgc aggctgctga gagagcactg 2760 tccagtgtga gatcaatgct tggtcaacct ctgcataaac gacaaggatc tccacgatca 2820 tttggtggga tgtcaaacaa gcgattaaag ccggactttc aacggtctct gcaacggatg 2880 ccatcttcgg gaagatactc ttaa 2904 <210> 19 <211> 2799 <212> DNA <213> Artificial Sequence <220> <223> CDS of Zea mays ZmCPL1.1 <400> 19 atgttcaagt cgatggttta ttacgtgaac acctcaatcg gagaggtgga ggtgtggccc 60 aagggcgagg cgagcgcggg cctgaccatg gcggcgtggg cgcgagaaat ccgcgttgaa 120 cgcatttccc cgcccagcga gcggtgccct ccgctggccg tcatgcacac cgtggccgtc 180 ggcgcccgat gcctcgtcat ggagtccagg ccgcccgttg tcgccgacgt tgtgccgcct 240 ctcgtcgtca tgcacaccgc ttgcctcagg gagaacaaga ctgcggttgt tccacttgga 300 gatgaagagt tgcatttagt tgcgatgaca tccagaagaa acttgacaaa tcatgcatgt 360 ttctggggct ataaattgcc atttggtttg tataattctt gcttgaccat gttaaatctt 420 cggtgcctgg gtattgtatt tgaccttgat gagacattga ttgtcgccaa tacatcacgg 480 tcttttgagg acagaattga cgcacttcaa agaaagctga gtaatgagac tgatccacaa 540 cgtaggaatg gtatgctatc agagatcaag aggtaccagg atgataagtc catcctaaag 600 caatatatag aaggtgatca ggtctatgat gatggaaaag tgtataaagc acaacctgag 660 attgttccac cattgtctga taaccagcaa ccaatgacac gtccagttat aagattacaa 720 gataaaaaca ttatcctgac aagaataaat cctctgatta gggataccag tgtgcttgta 780 tgtttaaggc cagcctggga ggatcttcgc agctacttaa ttgccagagg tcgcaagcgt 840 tttgaggtct atgtgtgtac gatggctgaa agagactatg ctttagaaat gtggagattg 900 cttgatccag attcaagatt gattaattct gttcaactcc atgatcggat ggtgtgtgta 960 aaatctggtt taaaaaagtc cttgctaaat gtcttccatg atggttcttg ccatcctggt 1020 atggcattag taattgatga tcgcctgaaa gtttgggatg agaaggatca attacgagtt 1080 catgtggttc ctgcatttac tccatattat gctcctcagg cggaggcaaa ttgttctatc 1140 ccagttctgt gtgtagccag aaacgttgca tgcaatgtta ggggtggttt cttcaaagac 1200 tttgatgaag gcctcttacc aaggattagc aatgttcact atgaggatga agtaaatgag 1260 atatctgcgc cagatgttgg caattatttg ataacagatg atgaaaatgt cgcattagtg 1320 aatgggaata gagattcatt gccttttgat ggtatggcag atgcagaggt tgagcggaga 1380 atgaaggaag ccaatgctca atcattccat caaacagcgg gagactttgt catgccagta 1440 gcccctgccc agaatttcgt ttcgacttca gttgcatcat tagccccacc tcttggcatg 1500 atgccatctc catttagcca gccagttgct ccaccaggtt tttcagattc actgcaaggt 1560 tctccagcta gagaagaggg tgaagttcca gagtctgagt tggatccgga cacaaggaga 1620 aggcttctta tattacagca tggccaagac acaagagatc ctacatctcc actaccagca 1680 ataccacccg tccaagttcc agttcctcca gtgcaacccc atgggaattg gtttcccaca 1740 gaggatggga taaaccaaag taacctgaat agaggctcag caggattcac tgtagaatct 1800 gattctattg tctatgagaa aaagcaacca cctcatcctt cattctttca tggtggggat 1860 agtcctatgc cgtctgatag atttggctat cagaaccaga ggtttccctc tcagctacca 1920 cacgaggatc accccatgat gcagaaccat gcacctccaa aatacagatc cttttctggt 1980 gaggaactag catcttggca tgttccctca agccagagaa acaaccagat agaatcagga 2040 agacactttg cacaatatgc tgggacctct gctggcatat tagaggggat tgctctgaaa 2100 tgtggttcta aggtggagta caagtcagca ttatgtgata ctgcagaact acaattctct 2160 attgaggttt ggattgttgg ggaaaaggtc ggtgaaggaa ttggtaggac aaggagagaa 2220 gcacaacgcc aagctgcaga aatgtcttta agaaacttgg ccaataaata cttgtcatct 2280 gatccaaata agttgtctga tatgaaagaa aatgatttca gtagcaacag aaatgtcttt 2340 ggttactctg gaaatacgag ggatgatatg ttgccacttt caagtacttc cgaggaatct 2400 cgctttatga aaatggagaa caataattcc cggaaaacag gaagttctgt tgctgctctc 2460 aaagaacttt gcactgttga gggatataac ttagtttttc aagcctgtcc atcttcagca 2520 gatggtttag ttgggaaaga atcttatgct caggtacaag ttggtggaca aattctaggc 2580 aaaggagttg ggttaacatg ggaagaggcc aagctccagg ctgctgctga ggctcttgga 2640 actttaagat ccatgctagg tcaacttggt cataaacgat ctggctctcc aaggtcattg 2700 gcaccaaatt ttaacaagcg gttcaagcca gattttccaa ggaccgtaca aagagttcct 2760 tacggaacat attctaggat cgaaggtcat gttccttaa 2799 <210> 20 <211> 2805 <212> DNA <213> Artificial Sequence <220> <223> CDS of Zea mays ZmCPL1.2 <400> 20 atgttcaagt cgatggttta tttcgtgaac atctcaatcg gggaggttga ggtgtggccc 60 aagggccagg cgagcgcagg cctggccgtg gcggcgtggg cgcgagatat ccgcgttgac 120 cgcctttccc cgcccagcga gcggtgccct ccgctggccg tcatgcacac cgtggccgtc 180 gccgcccgat gcctcgtcat ggagtccagg ccgcccgtcg ccgccgacgt ggcgtcgctg 240 cctctcgtcg acatgcacgc cgcttgcctc agggacaaca agaccgcggt tgttgcactt 300 ggagatgaag aattgcattt agttgcgatg acatccagaa gaaacttgac aaatcatgca 360 tgtttctggg gctacaagtt gccatttggt ttgtataatt cttgcttgac catgttaaat 420 ctacggtgcc tgggaatcgt gtttgacctt gatgagacat tggttgtcgc caatacatca 480 cggtctttcg aggacagaat tgatgcactt caaagaaagc tgagcaatga gactgatcca 540 cagcgtagaa atggtatgct atcagagatc aagaggtacc aagatgataa atccatctta 600 aagcaatata tagaaggtga tcaggtctat gatgacggta aagtgtataa agcacaacct 660 gagattgttc caccattgtc tgataaccag caaacaatga cacgcccagt tataagatta 720 caagaaaaag acattatcct gacaagaata aatcctctga ttagggatac cagtgtgctt 780 gtacgtttaa ggccagcctg ggaggatctt cgcagctact taattgccag aggtcgcaaa 840 cgttttgagg tctatgtgtg tacgatggct gaaagagact atgctttaga aatgtggaga 900 ttgcttgatc cagattcaag attaattaat tctgttcgac ttcatgatag gatggtgtgc 960 gtaaaatctg gtttaaagaa gtccttgcta aatgtcttcc atgatggttc ttgtcatcct 1020 ggtatggcat tagtaattga tgatcgtctg aaagtttggg atgagaagga tcaattacga 1080 gtccatgtgg ttcctgcatt tactccatat tatgctccac aggcagaggc aaattgttct 1140 atcccggttc tttgtgtagc cagaaacgtt gcatgcaatg ttaggggtgg tttcttcaaa 1200 gacttcgatg aaggactctt accaaggatt agtaatgttc attatgaaga tgaagtaaat 1260 gatatctcat ctgcgccaga tgttggcaat tatttgataa cagaggatga gaatgtcgca 1320 ttagtgaatg gaaatagaga ttcattgcct tttgatggta tgtcagatgc agagattgaa 1380 cggagaatga aggaagccaa tgctcaagca ttccatcaaa cagcgactaa ctttgtcatg 1440 ccagtagccc ctgctcagaa ttttgtttcg tcttcagttg caccattagc gccacctctt 1500 agcatgatgc cgcctccatt cagccagcca gttgttcagc caggtttttc agatccactg 1560 caaggttctc cagctagaga agagggtgaa gttcctgagt ctgagttgga tccagacaca 1620 aggagaaggc ttctcatatt gcagcatggt caagacacaa gagatcctac acctccacac 1680 ccagcgatac caccagtgca agttccagtt cctccagtgc aacctcatgg gaattggttt 1740 cccacagagg atgggataaa cccaagtaac ctgagtagag gttcagcagg tttcaccgtt 1800 gaatccgatt ctatgcccta tgagaaaaaa caaccacctc atccttcatt ctttcatggt 1860 ggggatagtc ctatgtcgtc tgatagattt ggctatcaga accagaggtt tccctctcag 1920 ctaccgcaca ...

Claims

1. A method for increasing resistance and / or tolerance to fungal pathogens in maize (Zea mays) or wheat (Triticum aestivum) plants, plant parts or plant populations, comprising reducing or eliminating the expression and / or activity of ERF922 protein and / or gene in the plants, plant parts or plant populations, wherein the wild-type ERF922 protein consists of a sequence identical to any one of the sequences of SEQ ID NO: 37 or 43-48, or is encoded by a sequence identical to any one of the sequences of SEQ ID NO: 52 or 58-63, wherein the fungal pathogen in maize plants is Setosphaeria turcica, and the fungal pathogens in wheat plants are Fusarium graminearum or Zymoseptoria tritici.

2. The method according to claim 1, wherein the expression and / or activity of the ERF922 protein and / or gene is reduced or eliminated by knocking out the ERF922 gene or knocking down the ERF922 protein, and / or the expression and / or activity of the ERF922 protein and / or gene is reduced or eliminated by mutagenesis, RNAi or gene editing.

3. The method according to claim 2, wherein the method comprises introducing a mutation in the ERF922 gene or a nucleotide sequence of a gene encoding an ERF922 with a mutation in the genome of the plant or plant part, the mutation resulting in a reduction or elimination of the expression of the mRNA of the gene and / or the ERF922 protein, or a reduction in the post-translational activity of the ERF922 protein.

4. The method according to claim 3, wherein the method comprises: (a) introducing a mutation in the nucleotide sequence of the endogenous wild-type gene encoding ERF922 in the plant or plant part, resulting in a reduction or elimination of the expression of the endogenous full-length mRNA of the gene and / or the endogenous full-length ERF922 protein, or a reduction in the post-translational activity of the ERF922 protein; (b) introducing into the plant or plant part a nucleotide sequence encoding an ERF922 protein, an RNAi molecule targeting the nucleotide sequence encoding the ERF922 protein or hybridizing with the nucleotide sequence encoding the ERF922 protein, or a polynucleotide sequence encoding an RNAi molecule targeting the nucleotide sequence encoding the ERF922 protein or hybridizing with the nucleotide sequence encoding the ERF922 protein, or (c) introducing into the plant or plant part an RNA-specific or DNA-specific CRISPR / Cas system targeting or directed against the nucleotide sequence encoding the ERF922 protein, or one or more polynucleotide sequences encoding the RNA-specific CRISPR / Cas system.

5. A method for controlling fungal pathogen infection in maize and / or wheat populations, comprising: a) providing a plant produced by the method according to any one of claims 1-3, b) Cultivating the plant of a) under conditions of pathogen infection.

6. A method for producing feed or food having a reduced amount of fungal or bacterial toxins, comprising: A) Controlling pathogen infection in a plant population by the method of claim 5, B) Harvesting plant material from said population, and C) Producing feed or food from the harvested plant material.

7. Feed or food having a reduced amount of fungal or bacterial toxins obtained by the method according to claim 6.

8. A method for identifying maize and / or wheat plants, plant parts or plant populations having increased resistance and / or tolerance to fungal pathogens, comprising screening and / or identifying a mutation defined in any one of claims 1-3 in maize and / or wheat plants, plant parts or plant populations, or a reduced or eliminated expression and / or activity of the ERF922 protein and / or gene.

9. The method according to claim 8, further comprising the step of selecting a plant, plant part or plant population having a mutation defined in any one of claims 1-3, or selecting a plant, plant part or plant population having a reduced or eliminated expression and / or activity of ERF922.

Images