Maize plants producing enhanced resistance to northern leaf blight

Abstract

Provided herein are compositions and methods for producing maize plants that exhibit resistance to northern leaf blight. Isolated polynucleotides encoding polypeptides that confer resistance to northern leaf blight are provided, as are polynucleotide constructs comprising such polynucleotides, and maize plants comprising the polynucleotide constructs. The methods comprise expressing the isolated polynucleotides in maize cells via standard transformation methods and obtaining a maize plant from the maize cells.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 242,691, filed October 16, 2015, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This disclosure relates to useful compositions and methods for producing corn plants with enhanced resistance to northern leaf blight.

[0004] References to sequence lists submitted electronically

[0005] An official copy of this sequence list was submitted electronically via EFS-Web as an ASCII format sequence list, named "20160928_BB2396PCT_SequenceListing.txt", created on September 28, 2016, and is 65 kilobytes in size, and was submitted with this specification. The sequence list included in this ASCII format file is part of this specification and is incorporated herein by reference in its entirety. Background Technology

[0006] Northern leaf blight (NLB), induced by the fungal pathogen *Exserohilum turcicum* (formerly known as *Helminthosporlum turcicum*), is a severe leaf wilt disease of maize in many tropical and temperate environments. Symptoms can range from cigar-shaped lesions on the lower leaves to complete leaf destruction, reducing the amount of leaf surface area available for photosynthesis. This reduced photosynthetic capacity leads to a lack of carbohydrates required for grain filling, which affects grain yield. Mid-altitude tropical regions (approximately 900–1600 m above sea level) have a particularly favorable climate for NLB due to long dew periods and moderate temperatures. However, in temperate environments (such as the United States), NLB can also cause 30%–50% losses during the wet season, especially if infection is confirmed on the upper leaves of the plant during the silking stage.

[0007] The most effective and preferred method for controlling northern leaf blight is to cultivate resistant hybrids. Several varieties or races of maize leaf blight pathogen exist in nature, giving growers two hybrid choices: partially resistant hybrids, which provide low-level broad-spectrum protection against multiple races, and race-specific resistant hybrids, which provide protection against a specific race. The genetic origins of resistance to maize leaf blight pathogen have been described, and four maize leaf blight resistance loci have been identified: Ht1, Hr2, Ht3, and Htn1. The gene Ht1 is located on the long arm of chromosome 2, and is closely linked to the following: umc36 (Coe, EH et al. (1988), Corn and Corn Improvement, 3rd ed., pp. 81-258), sgcr506 (Gupta, M. et al. (1989), Maize Genet. Coop. Newsl., 63, 112), umc150B (Bentolila, S. et al. (1991), Theor. Appl. Genet., 82: 393-398), and pic18a (Collins et al. (1998), Molecular Plant-Microbe). Interactions [Molecular Plant-Microbe Interactions], 11: 968-978), and its immediate flanking sides are umc22 and umc122 (Li et al. (1998) Hereditas [Genetics], 129: 101-106). Gene Ht2 is located in the umc48-umc89 region on the long arm of chromosome 8 (Zaitlin et al. (1992) Maize Genet. Coop. Newsl. [Maize Genetics Collaboration Newsletter], 66, 69-70), and gene Ht3 is mapped to the vicinity of bnlg1666 on chromosome 7 (Van Staden, D et al. (2001) Maize Genetics Conference Abstracts [Maize Genetics Conference Abstracts] 43: P134). The Htn1 gene is located on chromosome 8, approximately 10 cM from Ht2 and 0.8 cM from the RFLP marker umc117 (Simcox and Bennetzen (1993) Maize Genet. Coop. Newl. [Corn Genetics Collaboration Newsletter] 67, 118-119; Simcox and Bennetzen (1993) Phytopathology [Plant Pathology], 83: 1326-1330).

[0008] Controlling northern leaf blight by reducing fungal inoculum requires additional time and resources from some farmers and may also have adverse environmental impacts. This makes the cultivation of resistant hybrids more attractive to farmers and the public. Therefore, it is desirable to provide compositions and methods for producing corn plants with enhanced resistance to northern leaf blight. Summary of the Invention

[0009] This article describes compositions and methods for producing corn plants that exhibit resistance to northern leaf blight, whether the resistance is newly conferred or enhanced.

[0010] This article introduces isolated polynucleotides that can be used to produce corn plants resistant to northern leaf blight. The isolated polynucleotide may be selected from the group consisting of: (a) the nucleotide sequence listed in SEQ ID NO: 1 (PH4GP c-DNA), SEQ ID NO: 3 (PH1W2 cDNA), or SEQ ID NO: 9 (PH4GP genomic sequence); (b) the nucleotide sequence encoding a CC-NB-LRR polypeptide having an amino acid sequence that, based on a CLUSTALW alignment using default parameters, has at least 90% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 4; (c) the nucleotide sequence encoding a CC-NB-LRR polypeptide having an amino acid sequence that, based on a CLUSTAL W alignment using default parameters, has at least 90% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 4, wherein the polypeptide comprises the amino acid sequence listed in SEQ ID NO: 10; and (d) the nucleotide sequence encoding a CC-NB-LRR polypeptide having the sequence listed in SEQ ID NO: 2 or SEQ ID NO: 4.

[0011] Also provided are polynucleotide constructs comprising isolated polynucleotides, wherein the isolated polynucleotides are efficiently linked to a promoter. The polynucleotide construct may further comprise one or more heterologous nucleic acid sequences encoding a polypeptide selected from the group consisting of: polypeptides conferring disease resistance, polypeptides conferring herbicide resistance, polypeptides conferring insect resistance, polypeptides involved in carbohydrate metabolism, polypeptides involved in fatty acid metabolism, polypeptides involved in amino acid metabolism, polypeptides involved in plant development, polypeptides involved in plant growth regulation, polypeptides involved in yield enhancement, polypeptides involved in drought resistance, polypeptides involved in cold resistance, polypeptides involved in heat resistance, and / or polypeptides involved in salt resistance, wherein the one or more heterologous nucleic acid sequences are efficiently linked to a promoter. For example, a polypeptide conferring disease resistance may be a polypeptide conferring resistance to Northern Leaf Blight (NLB), said polypeptide may further have an amino acid sequence having at least 90% sequence identity when compared to SEQ ID NO: 11 or 12 based on a CLUSTAL W alignment method using default parameters.

[0012] Also provided are corn plant cells containing these polynucleotide constructs and corn plants containing these corn plant cells.

[0013] This paper provides a method for producing maize plants resistant to northern leaf blight, comprising expressing a polynucleotide construct of an isolated polynucleotide provided herein ( wherein the isolated polynucleotide is efficiently linked to at least one regulatory sequence) in regenerable maize plant cells, and producing maize plants resistant to northern leaf blight from these maize plant cells. The maize plants produced by this method contain the polynucleotide construct in their genome. The regulatory sequence may be a promoter and / or a terminator, and may be natural to maize. In some respects, the regulatory sequence is natural to the Ht1 gene. In other respects, the polynucleotide construct comprises one or more additional heteronucleotide sequences encoding a polypeptide selected from the group consisting of: polypeptides conferring disease resistance, polypeptides conferring herbicide resistance, polypeptides conferring insect resistance, polypeptides involved in carbohydrate metabolism, polypeptides involved in fatty acid metabolism, polypeptides involved in amino acid metabolism, polypeptides involved in plant development, polypeptides involved in plant growth regulation, polypeptides involved in yield enhancement, polypeptides involved in drought resistance, polypeptides involved in cold resistance, polypeptides involved in heat resistance, and / or polypeptides involved in salt resistance, wherein each heteronucleotide sequence is effectively linked to a promoter. The polypeptide may be a polypeptide conferring resistance to northern leaf blight (NLB), such as a polypeptide having an amino acid sequence with at least 90% sequence identity when compared to SEQ ID NO: 11 or 12 using a CLUSTAL W alignment method with default parameters. Progeny plants containing the polynucleotide construct can also be produced by hybridizing a corn plant produced by this method with a second corn plant whose genome does not contain the polynucleotide construct.

[0014] Description of the attached figures and sequence listing

[0015] The invention will be more fully understood from the following detailed description, accompanying drawings, and sequence listing, which form part of this application. As defined by the IUPAC-IUBMB standard described in the following references, the sequence listing comprises single-letter codes for nucleotide sequence characters and three-letter codes for amino acids: Nucleic Acids Research 13:3021-3030 (1985) and Biochemical Journal 219(Vol. 2):345-373 (1984), which are incorporated herein by reference in their entirety. The notation and format used for nucleotide and amino acid sequence data follow the regulations as set forth in 37 C. FR § 1.822.

[0016] Figure 1A-1DThe comparison shows the CC-NB-LRR variants from PH4GP (SEQ ID NO: 2), PH1W2 (SEQ ID NO: 4), and B73 (SEQ ID NO: 6 and 8). Figure 1C The deletion of the LRR region in the B73 allele is shown in the middle box.

[0017] SEQ ID NO: 1 is the nucleotide sequence of Ht1 cDNA found in the inbred line PH4GP.

[0018] SEQ ID NO: 2 is the amino acid sequence of the polypeptide encoded by SEQ ID NO: 1.

[0019] SEQ ID NO: 3 is the nucleotide sequence of Ht1 cDNA found in the inbred line PH1W2.

[0020] SEQ ID NO:4 is the amino acid sequence of the polypeptide encoded by SEQ ID NO:3.

[0021] SEQ ID NO: 5 is the nucleotide sequence of Ht1 cDNA found in the inbred line B73 and is referred to herein as “B73-high allele”.

[0022] SEQ ID NO: 6 is the amino acid sequence of the polypeptide encoded by SEQ ID NO: 5.

[0023] SEQ ID NO: 7 is the nucleotide sequence of Ht1 cDNA found in the inbred line B73 and is referred to herein as “B73-low allele”.

[0024] SEQ ID NO: 8 is the amino acid sequence of the polypeptide encoded by SEQ ID NO: 7.

[0025] SEQ ID NO: 9 is the nucleotide sequence of the Ht1 genomic DNA found in the inbred line PH4GP.

[0026] SEQ ID NO: 10 is an amino acid sequence of a region found in the Ht1 polypeptide of the resistance allele.

[0027] SEQ ID NO: 11 is the amino acid sequence of NLB18 from strain PH99N in patent application WO 2011163590.

[0028] SEQ ID NO: 12 is the amino acid sequence of NLB18 from strain PH26N in patent application WO 2011163590. Detailed Implementation

[0029] Before describing the invention in detail, it should be understood that the invention is not limited to the specific embodiments, which can of course vary. It should also be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be restrictive. When used in this specification and the appended claims, singular and singular terms such as “a / an” and “the” include plural referents unless explicitly indicated by the context. Thus, for example, references to “plant,” “the plant,” or “a type of plant” also include multiple plants; additionally, depending on the context, the use of the term “plant” may also include genetically similar or identical offspring of that plant; in fact, the use of the term “nucleic acid” optionally includes many copies of the nucleic acid molecule; similarly, the term “probe” optionally (and typically) covers many similar or identical probe molecules.

[0030] Unless otherwise specified, nucleic acids are written from left to right in a 5′ to 3′ direction. Numerical ranges described in the specification include numbers within defined ranges and include every integer or any non-integer fraction within those ranges. Unless otherwise specified, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar to or equivalent to those described herein may be used in the practice of testing the invention, preferred materials and methods are described herein. In describing and claiming protection for this invention, the following terms will be used according to the definitions stated below.

[0031] I. Composition

[0032] A.Ht1 polynucleotides and peptides

[0033] US 2010095395 describes the mapping of a QTL associated with resistance to northern leaf blight on chromosome 2 using a population derived from a cross between the northern leaf blight resistant strain PH4GP and the northern leaf blight susceptible strain PH5W4. This paper presents the cloning of the Ht1 gene in maize and the identification of the putative CC-NB-LRR (coil-and-coil, nucleotide-bound, leucine-rich repeat) gene as a pathogenic gene. The Ht1 cDNA sequences from the two resistance sources described in US 2010095395, PH4GP and PH1W2, are represented by SEQ ID NO: 1 and 3, respectively, while the amino acid sequences of the encoded polypeptides are represented by SEQ ID NO: 2 and 4. Furthermore, a construct containing the PH4GP (resistance) allele (SEQ ID NO: 9) was generated and transformed into susceptible transformants using Agrobacterium-mediated transformation, resulting in maize plants resistant to northern leaf blight.

[0034] The Zea mays CC-NB-LRR (coil-coiled, nucleotide-bound, leucine-rich repeat; also known as Ht1) gene is a member of a large, complex family of disease resistance genes. The mechanisms of NB-LRR protein activation and subsequent signal transduction in effector-triggered immunity are not fully understood (Eitas and Dangl. 2010. Curr Opin PlantBiol [New Insights in Plant Biology] 13(4): 472-477).

[0035] Therefore, this paper introduces a polynucleotide that can be used to produce corn plants resistant to northern leaf blight. This polynucleotide can be: (a) a nucleotide sequence listed in SEQ ID NO: 1 (PH4GP c-DNA), SEQ ID NO: 3 (PH1W2 cDNA), or SEQ ID NO: 9 (PH4GP genomic sequence); (b) a nucleotide sequence encoding a CC-NB-LRR polypeptide having an amino acid sequence that, based on a CLUSTAL W alignment using default parameters, has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 4; or (c) a nucleotide sequence encoding a CC-NB-LRR polypeptide having an amino acid sequence that, based on a CLUSTAL W alignment using default parameters, has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 4. Compared to NO:4, the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, wherein the polypeptide comprises the amino acid sequence listed in SEQ ID NO:10, which is the sequence of the Ht1 deletion region in B73; and (d) the nucleotide sequence encoding the CC-NB-LRR polypeptide, wherein the polypeptide has the sequence listed in SEQ ID NO:2 or SEQ ID NO:4.

[0036] The use of the term "polynucleotide" is not intended to limit the polynucleotides disclosed herein to polynucleotides containing DNA. Those skilled in the art will recognize that polynucleotides can comprise ribonucleotides and combinations of ribonucleotides and deoxyribonucleotides. Such deoxyribonucleotides and ribonucleotides include both naturally occurring molecules and synthetic analogs. The polynucleotides disclosed herein also encompass all forms of sequences, including but not limited to single-stranded, double-stranded, hairpin, stem-loop, and so on.

[0037] As used herein, “isolated” or “purified” polynucleotides or polypeptides, or their bioactive portions, are substantially or essentially free of components that normally accompany or interact with polynucleotides or polypeptides found in their natural environment. Therefore, isolated or purified polynucleotides or polypeptides are substantially free of other cellular material or culture media when produced by recombinant technology, or substantially free of chemical precursors or other chemicals when chemically synthesized. Preferably, the “isolated” polynucleotide does not contain sequences naturally flanking the polynucleotide in the genomic DNA of the organism from which it is derived (i.e., sequences located at the 5′ and 3′ ends of the polynucleotide) (preferably protein-coding sequences). For the purposes of this disclosure, “isolated” or “recombinant” when used to refer to nucleic acid molecules does not include isolated, unmodified chromosomes. For example, in various embodiments, the isolated polynucleotide may contain nucleotide sequences less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb, which are naturally located flanking the polynucleotide in the genomic DNA of the cell from which it is derived. Peptides that are essentially free of cellular material include formulations of peptides having less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of contaminating proteins. When recombinantly producing the peptides disclosed herein or their bioactive portions, the culture medium preferably represents less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of chemical precursors or non-target protein chemicals.

[0038] As used herein, a “recombinant” polynucleotide comprises a combination of two or more chemically linked nucleic acid segments (which are not found to be directly linked in nature). “Directly linked” is intended to mean that two nucleic acid segments are closely adjacent and linked to each other by a chemical bond. In a specific embodiment, the recombinant polynucleotide comprises a target polynucleotide such that the additional chemically linked nucleic acid segment is located at the 5′, 3′, or inside of the target polynucleotide. Alternatively, the chemically linked nucleic acid segment of the recombinant polynucleotide can be formed by deleting sequences. The additional chemically linked nucleic acid segment, or the sequence deleted to link the linked nucleic acid segments, can have any length, including, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or more nucleotides. Various methods for producing such recombinant polynucleotides are disclosed herein, including, for example, by chemical synthesis or by manipulating isolated polynucleotide segments using genetic engineering techniques. In a specific embodiment, the recombinant polynucleotide may comprise a recombinant DNA sequence or a recombinant RNA sequence.

[0039] A “recombinant polypeptide” comprises a combination of two or more chemically linked amino acid segments (which are not found to be directly linked in nature). In a specific embodiment, the recombinant polypeptide comprises additional chemically linked amino acid segments located at or within the N-terminus, C-terminus, or interior of the recombinant polypeptide. Alternatively, the chemically linked amino acid segments of the recombinant polypeptide can be formed by deleting at least one amino acid. The additional chemically linked amino acid segments or the deleted chemically linked amino acid segments can have any length, including, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or more amino acids.

[0040] Sequence alignment and identity percentage calculation can be determined using a variety of comparison methods designed for detecting homologous sequences, including but not limited to... Bioinformatics Computing Package ( The company (Madison, Wisconsin) Procedure. Unless otherwise stated, the multiple alignments of sequences provided herein were performed using the CLUSTAL V alignment method (Higgins and Sharp, CABIOS. [Computers in Biology] 5: 151 153 (1989)) and default parameters (vacancy penalty = 10, vacancy length penalty = 10). The default parameters for step-by-step alignment and calculation of the percentage of identity for protein sequences using the CLUSTAL V method are KTUPLE = 1, vacancy penalty = 3, window (WINDOW) = 5, and stored diagonals (DIAGONALS SAVED) = 5. For nucleic acids, these parameters are KTUPLE = 2, vacancy penalty = 5, window = 4, and stored diagonals = 4. After aligning sequences using the CLUSTALW procedure, it is possible to obtain the "Percentage of Identity" and "Divergence" values ​​by consulting the "Sequence Distance" table in the same procedure. Unless otherwise stated, the percentage of identity and divergence provided and declared herein are calculated in this manner.

[0041] Alternatively, the Clustal W alignment method can be used. This Clustal W alignment method (described in Higgins and Sharp, CABIOS. [Computers in Biology] 5: 151-153 (1989); Higgins, DG et al., Comput. Appl. Biosci. [Computers in Biological Sciences] 8: 189-191 (1992)) can be used... Bioinformatics Computing Package ( MegAlign, Madison, Wisconsin TMThe default parameters for multiple alignments are found in program v6.1. These parameters correspond to a gap penalty of 10, a gap length penalty of 0.2, a delayed divergent sequence of 30%, a DNA conversion weight of 0.5, a protein weight matrix of Gonnet series, and a DNA weight matrix of IUB. For pairwise alignments, the default parameters are alignment = Slow-Accurate, a gap penalty of 10.0, a gap length of 0.10, a protein weight matrix of Gonnet 250, and a DNA weight matrix of IUB. After aligning sequences using the Clustal W program, it is possible to obtain the "Percentage of Identity" and "Divergence" values ​​by consulting the "Sequence Distance" table in the same program.

[0042] B. Polynucleotide constructs

[0043] The Ht1 polynucleotide disclosed herein can be provided as an expression cassette (e.g., in the form of a polynucleotide construct) for expression in a target plant or any target organism. The cassette may include 5′ and 3′ regulatory sequences efficiently linked to the Ht1 polynucleotide. “Efficiently linked” is intended to indicate a functional link between two or more elements. For example, an efficient link between the target polynucleotide and a regulatory sequence (i.e., a promoter) is a functional link that allows expression of the target polynucleotide. The efficiently linked elements can be contiguous or non-contiguous. When used to refer to the link between two protein-coding regions, an efficient link is intended to mean that these coding regions are in the same reading frame. The cassette may additionally contain at least one additional gene to be co-transformed into an organism. Alternatively, the one or more additional genes may be provided on multiple expression cassettes. Such an expression cassette is equipped with multiple restriction sites and / or recombination sites for inserting the Ht1 polynucleotide under transcriptional regulation of the regulatory region. The expression cassette may additionally contain a selective marker gene.

[0044] This expression cassette contains, in a 5'-3' transcriptional orientation, transcription and translation initiation regions (i.e., promoters), Ht1 polynucleotides, and transcription and translation termination regions that function in plants (i.e., termination regions). Regulatory regions (i.e., promoters, transcription regulatory regions, and translation termination regions) and / or Ht1 polynucleotides may be native to or similar to each other in maize plant cells. Alternatively, regulatory regions and / or Ht1 polynucleotides may be heterologous to or to each other in maize plant cells.

[0045] As used herein, “heterologous” in the context of a sequence means that the sequence originates from a foreign species, or, if from the same species, is a sequence substantially modified from its natural form in the composition and / or genomic locus through deliberate human intervention. For example, a promoter effectively linked to a heterologous polynucleotide originates from a species different from the species from which the polynucleotide is derived, or, if from the same / similar species, one or both are substantially modified from their original form and / or genomic locus, or the promoter is not a natural promoter of the polynucleotide being effectively linked.

[0046] The termination region may be natural with the transcription start region, natural with the corn plant, or derived from another source (i.e., the promoter, Ht1 polynucleotide, corn plant, or any combination thereof may be exogenous or heterologous).

[0047] The expression cassette may additionally contain a 5′ leader sequence. Such leader sequences can enhance translation. Translational leaders are known in the art and include viral translational leader sequences.

[0048] In preparing expression cassettes, various DNA fragments can be manipulated to provide DNA sequences that are in the appropriate orientation and, at the appropriate time, within the appropriate reading frame. For this purpose, adapters or linkers can be used to ligate the DNA fragments, or other manipulations can be involved to provide convenient restriction sites, remove redundant DNA, and remove restriction sites. For this purpose, in vitro mutagenesis, primer repair, restriction enzyme digestion, annealing, and substitution (e.g., conversion and transversion) can be employed.

[0049] Many promoters can be used to express the various Ht1 sequences disclosed herein, including natural promoters of the target polynucleotide sequence (e.g., the natural promoter of the Ht1 gene). These promoters can be selected based on the desired outcome. Such promoters include, for example, constitutive promoters, inducible promoters, tissue-preferred promoters, or other promoters for expression in plants or any target organism. Synthetic promoters can also be used to express Ht1 sequences. Synthetic promoters include, for example, combinations of one or more heterologous regulatory elements.

[0050] Polynucleotide constructs can be recombinant DNA constructs. A “recombinant DNA construct” comprises two or more efficiently linked DNA segments that are not found to be efficiently linked in nature. Non-limiting examples of recombinant DNA constructs include target polynucleotides efficiently linked to heterologous sequences that facilitate the expression, autonomous replication, and / or genomic insertion of the target sequence. Such heterologous and efficiently linked sequences include, for example, promoters, terminator sequences, enhancers, etc., or any component of an expression cassette; plasmids, kinases, viruses, autonomously replicating sequences, bacteriophages, or linear or circular single-stranded or double-stranded DNA or RNA nucleotide sequences; and / or sequences encoding heterologous polypeptides.

[0051] C. Maize plant cells and maize plants

[0052] "Maize" refers to the maize plant (Zea mays L.ssp.mays), which is also known as "corn".

[0053] Also provided are maize plants, maize plant cells, maize plant parts and seeds, and maize grains having the Ht1 sequence disclosed herein. In specific embodiments, the plant and / or plant parts have been stably incorporated into at least one heterologous Ht1 polypeptide disclosed herein. Additionally, the target plant or organism may contain multiple Ht1 polynucleotides (i.e., at least 1, 2, 3, 4, 5, 6, or more).

[0054] As used herein, the term maize plant includes maize plant cells, maize plant protoplasts, maize plant cell tissue cultures from which maize plants can regenerate, maize plant callus, maize plant masses, and intact maize plant cells in maize plants or maize plant parts (such as embryos, pollen, ovules, seeds, leaves, flowers, kernels, spikes, rachis, husks, stems, roots, root tips, anthers, etc.). Grain refers to mature seeds produced by commercial growers for purposes other than cultivation or propagation of the species. D. Other purposeful traits

[0055] In some embodiments, the Ht1 polynucleotide disclosed herein can be engineered into molecular stacks. Therefore, the various maize plants, maize plant cells, and maize seeds disclosed herein can further comprise one or more desired traits, and in more specific embodiments, the maize plant, maize plant portion, or maize plant cell is stacked with any combination of the desired polynucleotide sequence to produce a plant having a desired combination of traits.

[0056] As used herein, the term "stack" includes the presence of multiple traits in the same target plant or organism. In a non-limiting instance, "stacked trait" includes a molecular stack in which sequences are physically adjacent to each other. As used herein, a trait refers to a phenotype derived from a particular sequence or group of sequences.

[0057] The polynucleotide DNA constructs described herein may also include one or more heteronucleotide sequences encoding polypeptides selected from the group consisting of: polypeptides conferring disease resistance, polypeptides conferring herbicide resistance, polypeptides conferring insect resistance, polypeptides involved in carbohydrate metabolism, polypeptides involved in fatty acid metabolism, polypeptides involved in amino acid metabolism, polypeptides involved in plant development, polypeptides involved in plant growth regulation, polypeptides involved in yield enhancement, polypeptides involved in drought resistance, polypeptides involved in cold resistance, polypeptides involved in heat resistance, and / or polypeptides involved in salt resistance, wherein each heteronucleotide sequence is effectively linked to a promoter.

[0058] The resistant peptide can be another peptide that confers resistance to northern leaf blight (NLB). For example, a polynucleotide DNA construct can contain the resistance allele for Ht1 and the resistance allele for NLB18 (in WO 2011163590). The amino acid sequence of the NLB18 peptide from strain PH99N is presented herein as SEQ ID NO: 11; the amino acid sequence of the NLB18 peptide from strain PH26N is presented herein as SEQ ID NO: 12. As described in application WO 2011163590, both PH99N and PH26N are corn strains that have shown resistance to northern leaf blight, reflecting different sources of resistance regarding the QTL on chromosome 8. The resistance allele of NLB18 can encode a polypeptide having an amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99% sequence identity when compared with SEQ ID NO: 11 or 12 based on the CLUSTAL W alignment method using default parameters.

[0059] II. Methods for producing corn plants resistant to northern leaf blight

[0060] "Exserohilum turcicum," formerly known as Helminthosporium turcicum, is a fungal pathogen that induces leaf blight infection in northern maize. This fungal pathogen is also referred to in this article as Exserohilum or Et.

[0061] "Disease resistance" (e.g., resistance to northern leaf blight) is a characteristic of a plant that avoids disease symptoms as a result of plant-pathogen interactions (e.g., the interaction between corn and maize large leaf spot fungus). That is, preventing pathogens from causing plant diseases and associated disease symptoms, or alternatively, minimizing or mitigating disease symptoms caused by pathogens. Those skilled in the art will understand that the compositions and methods disclosed herein can be used in conjunction with other compositions and methods available in the art for protecting plants from pathogen attack.

[0062] "Resistance" is a relative term that indicates that an infected plant produces a better corn yield than another similarly treated, more susceptible plant. That is, the disease results in a reduction in corn survival and / or yield in tolerant corn plants compared to susceptible corn plants. Those skilled in the art will understand that corn plants resistant to Northern Leaf Blight or the pathogen that causes it can represent a range of more or less resistant phenotypes and can vary depending on the severity of the infection. However, with simple observation, those skilled in the art can determine the relative resistance or susceptibility of different plants, plant lines, or plant families to Northern Leaf Blight and will also identify phenotypic grades of "resistance." For example, a visual scale of 1 to 9 can be used to represent the level of resistance to Northern Leaf Blight. Higher scores indicate higher resistance. Data should only be collected if sufficient selection pressure exists in the experiment being measured. The terms "tolerance" and "resistance" are used interchangeably herein.

[0063] Resistance can be “newly conferred” or “enhanced.” “Newly conferred” or “enhanced” resistance refers to an increased level of resistance against a specific pathogen, a broad-spectrum pathogen, or an infection caused by one or more pathogens. For example, an increased level of resistance against a specific fungal pathogen (such as Et) constitutes “enhanced” or improved fungal resistance. Embodiments of the present invention will enhance or improve resistance to fungal plant pathogens, thereby increasing the plant’s resistance to one or more fungal pathogens, which in turn increases resistance to diseases caused by fungal pathogens. The term “enhanced” means improved, increased, expanded, multiplied, elevated, or improved.

[0064] The corn plants produced by the methods described herein can provide durable and broad-spectrum resistance to corn and can assist in the breeding of northern leaf blight resistant corn plants. For example, if multiple northern leaf blight resistance genes are stacked into a single unit, this reduces the number of specific loci that need to be introduced through backcrossing and minimizes linkage redundancy from non-superior resistance donors.

[0065] Various methods can be used to introduce a target sequence into corn plant cells, corn plants, or corn plant parts. "Introduction" is intended to mean providing a polynucleotide to a corn plant cell, corn plant, or corn plant part in such a manner that the sequence enters the interior of the corn plant cell. The methods disclosed herein do not rely on a specific method for introducing the sequence into an organism or corn plant or corn plant part, as long as the polynucleotide enters the interior of at least one cell of the corn plant. Methods for introducing polynucleotides into various organisms (including corn plants) are known in the art, including but not limited to stable transformation, transient transformation, and virus-mediated transformation.

[0066] "Stable transformation" refers to the integration of a polynucleotide construct introduced into a maize plant into the maize plant's genome, which is then inherited by its offspring. "Transient transformation" refers to the introduction of a polynucleotide into a maize plant that does not integrate into the maize plant's genome.

[0067] Transformation protocols and protocols for introducing polynucleotide sequences into plants (e.g., corn) can be modified. Suitable methods for introducing polynucleotides into corn plant cells include microinjection (Crossway et al. (1986) Biotechniques [Biotechnology] 4: 320 334), electroporation (Riggs et al. (1986) Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences of the United States of America] 83: 5602 5606), Agrobacterium-mediated transformation (US Patent Nos. 5,563,055 and 5,981,840), direct gene transfer (Paszkowski et al. (1984) EMBOJ. [Journal of the European Society for Molecular Biology] 3: 2717 2722), and ballistic particle acceleration (see, for example, US Patent Nos. 4,945,050; US Patent Nos. 5,879,918; US Patent Nos. 5,886,244; and 5,932,782; Tomes et al. (1995), Plant cell, Tissue, and Organ Culture: Fundamental Methods, edited by Gamborg and Phillips (Springer Publishing, Berlin); McCabe et al. (1988) Biotechnology 6: 923-926; and Lec1 transformation method (WO 00 / 28058).See also Klein et al. (1988) Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences of the United States of America] 85: 4305 4309 (Corn); Klein et al. (1988) Biotechnology 6: 559 563 (Corn); US Patent Nos. 5,240,855; 5,322,783; and 5,324,646; Klein et al. (1988) Plant Physiol. 91: 440 444 (Corn); Fromm et al. (1990) Biotechnology 8: 833 839 (Corn); Hooykaas-Van Slogteren et al. (1984) Nature (London) 311: 763-764; US Patent No. 5,736,369 (Cereals); DeWet et al. (1985) In The Experimental Manipulation of Ovule Tissues, edited by Chapman et al. (Longman, New York), pp. 197-209 (Pollen); D'Halluin et al. (1992) Plant Cell 4: 1495-1505 (Electroporation); and Osjoda et al. (1996) Nature Biotechnology 14: 745-750 (Corn transformed by Agrobacterium tumefaciens); all of which are incorporated herein by reference.

[0068] In other embodiments, the Ht1 polynucleotide disclosed herein can be introduced into plants by contacting the plant with a virus or viral nucleic acid. Typically, such methods involve incorporating the disclosed polynucleotide construct into a DNA or RNA molecule. It should be appreciated that the Ht1 sequence can initially be synthesized as part of a viral polyprotein and then processed by in vivo or in vitro proteolysis to produce the desired recombinant protein. Furthermore, it should be appreciated that the promoters disclosed herein also cover promoters for transcription via viral RNA polymerase. Methods involving viral DNA or RNA molecules for introducing polynucleotides into plants and expressing the proteins encoded therein are known in the art. See, for example, U.S. Patent Nos. 5,889,191, 5,889,190, 5,866,785, 5,589,367, 5,316,931 and Porta et al. (1996) Molecular Biotechnology 5:209-221; these documents are incorporated herein by reference.

[0069] Methods for targeted insertion of polynucleotides at specific locations in a plant genome are known in the art. In one embodiment, the insertion of a polynucleotide at a desired genomic location is achieved using a site-specific recombination system. See, for example, WO 99 / 25821, WO 99 / 25854, WO 99 / 25840, WO 99 / 25855, and WO 99 / 25853, all of which are incorporated herein by reference. Briefly, the polynucleotides disclosed herein can be contained in a transfer cassette flanked by two non-recombination-inducing recombination sites. The transfer cassette is introduced into a plant to stably incorporate a target site into its genome, flanked by two non-recombination-inducing recombination sites corresponding to those sites on the transfer cassette. A suitable recombinase is provided, and the transfer cassette is integrated into the target site. Thus, the target polynucleotide is integrated at a specific chromosomal location in the plant genome. Other approaches to targeting polynucleotides are described in WO 2009 / 114321 (incorporated hereby by reference), which describes the generation of “custom-made” large-scale nucleases to modify plant genomes, particularly the corn genome. See also Gao et al. (2010) Plant Journal 1: 176-187.

[0070] Transformed cells can be cultured into plants using conventional methods. See, for example, McCormick et al. (1986) Plant Cell Reports 5: 81-84. These plants can then be cultured and pollinated with the same or different transformed lines, and the resulting progeny with constitutive expression of the desired phenotypic trait can be identified. Two or more generations of plants can be cultured to ensure that the expression of the desired phenotypic trait is stably maintained and inherited, and the seeds are then harvested to ensure that the expression of the desired phenotypic trait has been achieved. In this way, this disclosure provides transformed seeds (also known as “transgenic seeds”) having the polynucleotides disclosed herein, which are stably incorporated into their genome, for example, as part of an expression cassette.

[0071] Transformed maize plant cells derived through plant transformation techniques (including those discussed above) can be cultured to regenerate complete plants possessing the transformed genotype (i.e., the Ht1 polynucleotide encoding a polypeptide conferring resistance to northern leaf blight) and thus the desired phenotype (e.g., resistance to northern leaf blight, whether newly conferred or enhanced). For transformation and regeneration of maize, see Gordon-Kamm et al., The Plant Cell, 2: 603-618 (1990). Plant regeneration from cultured protoplasts is described in Evans et al. (1983), *Protoplasts Isolation and Culture, Handbook of Plant Cell Culture*, pp. 124–176, Macmillan Publishing Company, New York; and Binding (1985), *Regeneration of Plants, Plant Protoplasts*, pp. 21–73, CRC Press, Pocaraton. Regeneration can also be achieved from plant callus, explants, organs, or parts thereof. Such regeneration techniques are generally described in Klee et al. (1987), *Annals of Plant Physiology*, 38:467. See also Payne and Gamborg.

[0072] Technicians will recognize that once the expression cassette containing the Ht1 gene is stably incorporated into transgenic plants and proven effective, it can be introduced into other plants through sexual hybridization. Any of many standard breeding techniques can be used, depending on the species to be hybridized.

[0073] In some embodiments, the method includes introduction via expression of a polynucleotide construct in regenerable maize plant cells, the polynucleotide construct comprising a polynucleotide efficiently linked to at least one regulatory sequence, wherein the polynucleotide encodes a resistance allele of the Ht1 gene described herein, and producing maize plants resistant to northern leaf blight from the maize plant cells. The maize plants produced by this method contain the polynucleotide construct in their genome. The regulatory sequence may be a promoter and / or a terminator and may be natural to the maize. In some embodiments, the regulatory sequence is natural to the Ht1 gene. Progeny plants containing the polynucleotide construct can also be produced by crossing the maize plants produced by this method with a second maize plant whose genome does not contain the polynucleotide construct. In some embodiments, the Ht1 gene is overexpressed (as a genomic fragment or cDNA) to confer higher resistance than the expression level in the natural state.

[0074] Example

[0075] The following examples are provided to illustrate, but do not limit, the appended claims. It should be understood that the examples and embodiments described herein are for illustrative purposes only, and those skilled in the art will recognize that various reagents or parameters can be changed without departing from the spirit of the invention or the scope of the appended claims.

[0076] Example 1

[0077] Fine mapping of the resistance locus for leaf blight in northern China

[0078] To finely map the Northern Leaf Blight resistance QTL located on chromosome 2, a large backcross-derived population was created. This population was created from a hybrid between the resistant line PH4GP (score = 9) and the susceptible line PH5W4 (score = 1), with the susceptible line used as the recurrent parent. Northern Leaf Blight infection scores were assigned to BC5 individuals using the phenotypic method disclosed in U.S. Patent 8,921,646. Marker recombination data from a large number of individuals in population 2 placed genes in an 18kb region of the B73 genome containing the putative protein phosphatase 2C, the putative PHD-type zinc finger protein, and the putative resistance protein. No known EST was found for this putative resistance protein; furthermore, based on the predicted gene sequence of B73, this putative resistance protein exhibited low constitutive expression.

[0079] Example 2

[0080] Identification of candidate genes for Ht1 and comparison of allele variants

[0081] A BAC library from the resistance donor PH4GP was constructed, and BAC clones covering the Ht1 region were identified and sequenced. The Ht1 region in PH4GP is less than 10 kb and contains only one annotation gene encoding the putative CC-NB-LRR (coil-and-coil, nucleotide-bound, leucine-rich repeat) gene. The Ht1 cDNA sequences from PH4GP and PH1W2 (another source of the resistance allele of Ht1; US ​​2010095395) are represented by SEQ ID NO: 1 and 3, respectively, while the amino acid sequences of the encoded polypeptides are represented by SEQ ID NO: 2 and 4. B73 has two splicing variants, and this novel variant (referred to herein as B73-high) is expressed at a much higher level than the known variant (referred to herein as B73-low). SEQ ID NO: 5 is the cDNA sequence of the B73-high allele, while the amino acid sequence of the encoded polypeptide is represented by SEQ ID NO: 6. SEQ ID NO: 7 is the cDNA sequence of the B73-low allele, while the amino acid sequence of the encoded polypeptide is represented by SEQ ID NO: 8.

[0082] Figure 1A-1D The CC and NB domains showed high similarity between the susceptibility allele (from B73) and the resistance allele (from PH4GP and PH1W2). However, both B73 alleles were absent in LRR. Figure 1C (as shown in the box). The amino acid sequence of this region in the Ht1 resistance allele is represented by SEQ ID NO: 10.

[0083] Example 3

[0084] Genetically modified verification

[0085] A construct containing the PH4GP (resistance) allele (SEQ ID NO: 9) was generated and transformed into susceptible transforming lines using Agrobacterium-mediated transformation. This genomic sequence (SEQ ID NO: 9) contains a natural promoter, exons, introns, and terminator regions. Regenerated transgenic plants were grown in a greenhouse, and quantitative PCR analysis was performed to confirm the insertion of the T-DNA cassette containing the PH4GP (resistance) allele. Many events showed a single copy of the T-DNA insert, which was confirmed by qPCR using four flanking markers. Based on the marker data, 41 out of 50 events were positive for the insert, and 9 were negative (invalid).

[0086] The efficacy of all events against the northern leaf blight pathogen (Maize large leaf spot fungus) was tested in a greenhouse. First, all events were challenged against race 0 of the pathogen (for which the Ht1 gene is known to provide resistance); then these events were subjected to race 1 (for which Ht1 does not provide resistance). As determined by qPCR, all positive events were resistant to race 0, and all negative events were susceptible to race 0. As expected, all events were susceptible to race 1.

[0087] Example 4

[0088] Production of Northern Leaf Blight Resistant Maize Plants Expressing Maize Ht1 Peptide

[0089] For example, recombinant DNA-based transformation can be used to produce northern leaf blight resistant maize plants expressing the Ht1 gene. Recombinant DNA-based transformation methods are well known in the art, such as Agrobacterium tumefaciens-mediated transformation and particle bombardment-based transformation. For Agrobacterium tumefaciens-based plant transformation, vectors are constructed according to methods known in the art. These vectors contain a T-DNA insert having a promoter, introns, optional enhancers (such as a 35S enhancer element), an Ht1 variant DNA conferring resistance to northern leaf blight, and a plant terminator. Immature maize embryos are excised and infected with an Agrobacterium tumefaciens vector containing the target Ht1 variant. After infection, the embryos are transferred and cultured in a co-culture medium. After co-culture, the infected immature embryos are transferred to a culture medium to allow transgenic callus growth. PCR and optional Western blotting are used to sample the presumed transgenic callus to confirm the presence of the Ht1 variant gene. The presumed transgenic callus was held on a culture medium for further growth and selection prior to plant regeneration. During regeneration, the confirmed transgenic callus was transferred to a maturation medium and cultured for somatic embryo maturation. The mature embryos were then transferred to a regeneration medium for shoot and root formation. After shoots and roots emerged, individual plantlets were transferred to tubes with rooting medium. Plantlets with established shoots and roots were transplanted into pots in a greenhouse for further growth and to produce T1 seeds.

[0090] Furthermore, DNA constructs containing Ht1 variant DNA conferring resistance to northern leaf blight may also include another gene encoding a polypeptide conferring resistance to northern leaf blight, such as the gene encoding NLB18 (in WO 2011163590). As described in application WO 2011163590, both PH99N and PH26N are maize lines that exhibit resistance to northern leaf blight, reflecting different sources of resistance regarding the QTL on chromosome 8. The amino acid sequence of the NLB18 polypeptide from line PH99N is presented herein as SEQ ID NO: 11; the amino acid sequence of the NLB18 polypeptide from line PH26N is presented herein as SEQ ID NO: 12. The introduction of Ht1 and NLB18 into plants may have the effect of increasing resistance to race 0 of the maize large leaf spot pathogen, and / or may provide resistance to one or more races (e.g., race 1) to which other Ht1 genes do not confer resistance. sequence list <110> EI DuPont de Nemours Pioneer Hi-Bred International Hou, Zhenglin Leonard, April Li, Bailin Tabor, Girma <120> Developing corn plants with enhanced resistance to northern leaf blight. <130> BB2396WOPCT <140> PCT / US16 / 57081 <141> 2016-10-14 <150> US 62 / 242,691 <151> 2015-10-16 <160> 12 <170> PatentIn version 3.5 <210> 1 <211> 2658 <212> DNA <213> corn <400> 1 atggagaacc cagacgcgca ggcgaaggcg tgggcggcgg agatgcgcga gctggcctac 60 gacatggagg acagcatcga tctcttcacc caccacgtcg accacgaacc ggccgacacc 120 gccaccaccg gcgtcaagag gttcttctc cggatcatcc ggaagcttaa gaaactccac 180 taccgccaca ggtttgttca ggagatcaaa caactccacg accttgccaa cgaatcgtac 240 cggcgtagga agaggtacag gattgaggag ggcggttcaa gcctctcgca cgcggagatc 300 gatcctcggt tagaggcgct ctacgtggag gtggagaaac tcgtgggcat ccagggccca 360 agccaggaga tcattggaca gctcgtcggc gagaacgcag cggagcgacg gagggttgtc 420 gccgttgttg gatctggagg ttcaggcaag accacacttg ccaaacaggt gtacgagaaa 480 atcaggtgcc aattctcttg tgcagccttt gtgtctgtgt cgcaaaagcc caacatgaat 540 agcctcctgt gggagttgct atctcaaatc gggaaccatg gtggagattt aggaatgatg 600 gcagtaggat attgcagtga caaacaactg atcgacagac taagatcaca tcttgaaaag 660 cagaggtatc tcgttgtgat agatgatgtt tggacaaact cagcgtggga gaccatacaa 720 tgtgcgctcc ctaaaatgc ccatgcaagt aaataattc tgacaacacg aatcaacagt 780 gtaggccagt tctcctgcac tccagatgag ggttttatct atcagatgaa gcctctttgc 840 agaaacgatt ctgaaaatct gtttctgaaa aggacactat gtgataaaga taagtttcct 900 gctcagctgg aggggattaa aaacgagata atcgagaaat gcgatggttt gccactggct 960 attgttactc tagctagcat gttagctact aaacagagaa caagggaaga atgggagagg 1020 gcacttgatt caatccattc tatgcacaag aaagatagtg gcctggaagt gatggacaag 1080 atactgtctc tgagttacag ggatctacct cacaacatga gaaattgctt gctgtatctc 1140 agtacatttc cagaggacca cacgatttac aaagatgccc tagtatggag atggatggct 1200 gaagggttta tcgctgaaac acaaggcttt actttggagc aggttgccga gggctacttc 1260 tacgagtttg tgaacaggag tttggttcag cccataacct tgcgttcaag atatgaaatg 1320 cgtggagaag gaggttgccg agtccatgac attgtactga acttcctcat ctctcgtgca 1380 gctgaagaga actttttaac tacgctgtat ggcgcccagg gggttccatc ttcagaccga 1440 aggattcgcc ggctctctgt ctgggacagt ccagaacacg cactggcagt ctctagagcg 1500 accatgaatc tgtcccatct ccggtcagtt agaatatgca acgttggaga ctggcccgtg 1560 cctgctgttc tagacttacc tgtccttcga gtgttagatc tagagggatg ccgtgatctg 1620 aggatcgacg aacctgactg cattctaagc ttgtttcatc tgagatacct gggtttccgc 1680 agcgcaagtg gtgtcgtgct accggctcaa atcggaaatt tacaccatct gcagaccatc 1740 gatttaagcg ggactggagt gacacagctg ccagaaagca ttgtccagct caagcgactg 1800 atgcatcttg ttgggcaacg gctcatcatg ccagacgggt ttggtagcat ggaatccctt 1860 gaggagttag gtactatcga ctgctgcaag tgccccgtca gttttgggga agacctagca 1920 cttctgagca ggctgagggt gctccgagtg gctttcatcg gggtcgaaac aagtgacatg 1980 gaaaccagaa ggaaatcttt gatgtcatcc ctctgcaaac tcggaggaga caaccttcgg 2040 cgtgtcacta ttatcgacct cgctggcggt ggagattgct ttgtggagtc gtggcaccct 2100 cctcctcgtc tcctccagaa gttcatccat atcagtcagc aacagcactt ctccaggttt 2160 ccagaatgga tcagttcctg cctatgtgat ctcacccacc tggatataaa ggccgaaaag 2220 atggaaaggg agcatctaag tgttcttgaa cacctgcccg ccatccgttg cctatacctt 2280 ttcgtgaagc gagtctccga agacgggctc gccatcagcc acggcgcgtt ccgatgtcta 2340 cggcgtctcg agttctgcaa cgtagatgga cctggtttga tgtttgcagg aggcgttcca 2400 atgttggaat ggctgaggct cgggttcgac gcggatagag cgcaatcgac atacggcggt 2460 ctggaggttg gcatccagcg cctctcgtct ctcaaacatg tcgtgctcat tgtatggatg 2520 gtttctgaag gcggtgatga tccagcggag caagccgtct ggtctgccat caatggccaa 2580 gtagagatgc tccccaactc tccgacggtt gatatccggt ttcgtagacg gagtcagctg 2640 caggcaagct cagaataa 2658 <210> 2 <211> 885 <212> PRT <213> Zea mays <400> 2 Met Glu Asn Pro Asp Ala Gln Ala Lys Ala Trp Ala Ala Glu Met Arg 1 5 10 15 Glu Leu Ala Tyr Asp Met Glu Asp Ser Ile Asp Leu Phe Thr His His 20 25 30 Val Asp His Glu Pro Ala Asp Thr Ala Thr Thr Gly Val Lys Arg Phe 35 40 45 Phe Leu Arg Ile Ile Arg Lys Leu Lys Lys Leu His Tyr Arg His Arg 50 55 60 Phe Val Gln Glu Ile Lys Gln Leu His Asp Leu Ala Asn Glu Ser Tyr 65 70 75 80 Arg Arg Arg Lys Arg Tyr Arg Ile Glu Glu Gly Gly Ser Ser Leu Ser 85 90 95 His Ala Glu Ile Asp Pro Arg Leu Glu Ala Leu Tyr Val Glu Val Glu 100 105 110 Lys Leu Val Gly Ile Gln Gly Pro Ser Gln Glu Ile Ile Gly Gln Leu 115 120 125 Val Gly Glu Asn Ala Ala Glu Arg Arg Arg Val Val Ala Val Val Gly 130 135 140 Ser Gly Gly Ser Gly Lys Thr Thr Leu Ala Lys Gln Val Tyr Glu Lys 145 150 155 160 Ile Arg Cys Gln Phe Ser Cys Ala Ala Phe Val Ser Val Ser Gln Lys 165 170 175 Pro Asn Met Asn Ser Leu Leu Trp Glu Leu Leu Ser Gln Ile Gly Asn 180 185 190 His Gly Gly Asp Leu Gly Met Met Ala Val Gly Tyr Cys Ser Asp Lys 195 200 205 Gln Leu Ile Asp Arg Leu Arg Ser His Leu Glu Lys Gln Arg Tyr Leu 210 215 220 Val Val Ile Asp Asp Val Trp Thr Asn Ser Ala Trp Glu Thr Ile Gln 225 230 235 240 Cys Ala Leu Pro Lys Asn Ala His Ala Ser Lys Ile Ile Leu Thr Thr 245 250 255 Arg Ile Asn Ser Val Gly Gln Phe Ser Cys Thr Pro Asp Glu Gly Phe 260 265 270 Ile Tyr Gln Met Lys Pro Leu Cys Arg Asn Asp Ser Glu Asn Leu Phe 275 280 285 Leu Lys Arg Thr Leu Cys Asp Lys Asp Lys Phe Pro Ala Gln Leu Glu 290 295 300 Gly Ile Lys Asn Glu Ile Ile Glu Lys Cys Asp Gly Leu Pro Leu Ala 305 310 315 320 Ile Val Thr Leu Ala Ser Met Leu Ala Thr Lys Gln Arg Thr Arg Glu 325 330 335 Glu Trp Glu Arg Ala Leu Asp Ser Ile His Ser Met His Lys Lys Asp 340 345 350 Ser Gly Leu Glu Val Met Asp Lys Ile Leu Ser Leu Ser Tyr Arg Asp 355 360 365 Leu Pro His Asn Met Arg Asn Cys Leu Leu Tyr Leu Ser Thr Phe Pro 370 375 380 Glu Asp His Thr Ile Tyr Lys Asp Ala Leu Val Trp Arg Trp Met Ala 385 390 395 400 Glu Gly Phe Ile Ala Glu Thr Gln Gly Phe Thr Leu Glu Gln Val Ala 405 410 415 Glu Gly Tyr Phe Tyr Glu Phe Val Asn Arg Ser Leu Val Gln Pro Ile 420 425 430 Thr Leu Arg Ser Arg Tyr Glu Met Arg Gly Glu Gly Gly Cys Arg Val 435 440 445 His Asp Ile Val Leu Asn Phe Leu Ile Ser Arg Ala Ala Glu Glu Asn 450 455 460 Phe Leu Thr Thr Leu Tyr Gly Ala Gln Gly Val Pro Ser Ser Asp Arg 465 470 475 480 Arg Ile Arg Arg Leu Ser Val Trp Asp Ser Pro Glu His Ala Leu Ala 485 490 495 Val Ser Arg Ala Thr Met Asn Leu Ser His Leu Arg Ser Val Arg Ile 500 505 510 Cys Asn Val Gly Asp Trp Pro Val Pro Ala Val Leu Asp Leu Pro Val 515 520 525 Leu Arg Val Leu Asp Leu Glu Gly Cys Arg Asp Leu Arg Ile Asp Glu 530 535 540 Pro Asp Cys Ile Leu Ser Leu Phe His Leu Arg Tyr Leu Gly Phe Arg 545 550 555 560 Ser Ala Ser Gly Val Val Leu Pro Ala Gln Ile Gly Asn Leu His His 565 570 575 Leu Gln Thr Ile Asp Leu Ser Gly Thr Gly Val Thr Gln Leu Pro Glu 580 585 590 Ser Ile Val Gln Leu Lys Arg Leu Met His Leu Val Gly Gln Arg Leu 595 600 605 Ile Met Pro Asp Gly Phe Gly Ser Met Glu Ser Leu Glu Glu Leu Gly 610 615 620 Thr Ile Asp Cys Cys Lys Cys Pro Val Ser Phe Gly Glu Asp Leu Ala 625 630 635 640 Leu Leu Ser Arg Leu Arg Val Leu Arg Val Ala Phe Ile Gly Val Glu 645 650 655 Thr Ser Asp Met Glu Thr Arg Arg Lys Ser Leu Met Ser Ser Leu Cys 660 665 670 Lys Leu Gly Gly Asp Asn Leu Arg Arg Val Thr Ile Ile Asp Leu Ala 675 680 685 Gly Gly Gly Asp Cys Phe Val Glu Ser Trp His Pro Pro Pro Arg Leu 690 695 700 Leu Gln Lys Phe Ile His Ile Ser Gln Gln Gln His Phe Ser Arg Phe 705 710 715 720 Pro Glu Trp Ile Ser Ser Cys Leu Cys Asp Leu Thr His Leu Asp Ile 725 730 735 Lys Ala Glu Lys Met Glu Arg Glu His Leu Ser Val Leu Glu His Leu 740 745 750 Pro Ala Ile Arg Cys Leu Tyr Leu Phe Val Lys Arg Val Ser Glu Asp 755 760 765 Gly Leu Ala Ile Ser His Gly Ala Phe Arg Cys Leu Arg Arg Leu Glu 770 775 780 Phe Cys Asn Val Asp Gly Pro Gly Leu Met Phe Ala Gly Gly Val Pro 785 790 795 800 Met Leu Glu Trp Leu Arg Leu Gly Phe Asp Ala Asp Arg Ala Gln Ser 805 810 815 Thr Tyr Gly Gly Leu Glu Val Gly Ile Gln Arg Leu Ser Ser Leu Lys 820 825 830 His Val Val Leu Ile Val Trp Met Val Ser Glu Gly Gly Asp Asp Pro 835 840 845 Ala Glu Gln Ala Val Trp Ser Ala Ile Asn Gly Gln Val Glu Met Leu 850 855 860 Pro Asn Ser Pro Thr Val Asp Ile Arg Phe Arg Arg Arg Ser Gln Leu 865 870 875 880 Gln Ala Ser Ser Glu 885 <210> 3 <211> 2652 <212> DNA <213> Zea mays <400> 3 atggagaacc cagacgcgca ggcgaaggcg tgggcggcgg agatgcgcga gctggcctac 60 gacatggagg acagcatcga tctcttcacc caccacgtcg accacgaacc ggccgacacc 120 gccaccaccg gcgtcaagag gttcttcctc cggatcatcc ggaagcttaa gaaactccac 180 taccgccaca ggtttgttca ggagatcaaa caactccacg accttgccaa cgaatcgtac 240 cggcgtagga agaggtacag gattgaggag ggcggttcaa gcctctcgca cgcggagatc 300 gatcctcggt tagaggcgct ctacgtggag gtggagaaac tcgtgggcat ccagggccca 360 agccaggaga tcattggaca gctcgtcggc gagaacgcag cggagcgacg gagggttgtc 420 gccgttgttg gatctggagg ttcaggcaag accacacttg ccaaacaggt gtacgagaaa 480 atcaggtgcc aattctcttg tgcagccttt gtgtctgtgt cgcaaaagcc caacatgaat 540 agcctcctgt gggagttgct atctcaaatc gggaaccatg gtggagattt aggaatgatg 600 gcagtaggat attgcagtga caaacaactg atcgacagac taagatcaca tcttgaaaag 660 cagaggtatc tcgttgtgat agatgatgtt tggacaaact cagcgtggga gaccatacaa 720 tgtgcgctcc ctaaaatgc ccatgcaagt aaataattc tgacaacacg aatcaacagt 780 gtaggccagt tctcctgcac tccagatgag ggtttatct atcagatgaa gcctctttgc 840 agaaacgatt ctgaaaatct gtttctgaaa aggacactat gtgataaaga taagtttcct 900 gctcagctgg aggggattaa aaacgagata atcgagaaat gcgatggttt gccactggct 960 attgttactc tagctagcat gttagctact aaacagagaa caaggaaga atgggagagg 1020 gcacttgatt caatccattc tatgcacaag aaagatagtg gcctggaagt gatggacaag 1080 atactgtctc tgagttacag ggatctacct cacaacatga gaaattgctt gctgtatctc 1140 agtacatttc cagaggacca cacgatttac aaagatgccc tagtatggag atggatggct 1200 gaagggttta tcgctgaaac acaaggcttt actttggagc aggttgccga gggctacttc 1260 tacgagtttg tgaacaggag tttggttcag cccataacct tgcgttcaag atatgaaatg 1320 cgtggagaag gaggttgccg agtccatgac attgtactga acttcctcat ctctcgtgca 1380 gctgaagaga actttttaac tacgctgtat ggcgcccagg gggttccatc ttcagaccga 1440 aggattcgcc ggctctctgt ctgggacagt ccagaacacg cactggcagt ctctagagcg 1500 accatgaatc tgtcccatct ccggtcagtt agaatatgca acgttggaga ctggcccgtg 1560 cctgctgttc tagacttacc tgtccttcga gtgttagatc tagagggatg ccgtgatctg 1620 aggatcgacg aacctgactg cattctaagc ttgtttcatc tgagatacct gggtttccgc 1680 agcgcaagtg gtgtcgtgct accggctcaa atcggaaatt tacaccatct gcagaccatc 1740 gatttaagcg ggactggagt gacacagctg ccagaaagca ttgtccagct caagcgactg 1800 atgcatcttg ttgggcaacg gctcatcatg ccagacgggt ttggtagcat ggaatccctt 1860 gaggagttag gtactatcga ctgctgcaag tgccccgtca gttttgggga agacctagca 1920 cttctgagca ggctgagggt gctccgagtg gctttcatcg gggtcgaaac aagtgacatg 1980 gaaaccagaa ggaaatcttt gatgtcatcc ctctgcaaac tcggaggaga caaccttcgg 2040 cgtgtcacta ttatcgacct cgctggcggt ggagattgct ttgtggagtc gtggcaccct 2100 cctcctcgtc tcctccagaa gttcatccat atcagtcagc acttctccag gtttccagaa 2160 tggatcagtt cctgcctatg tgatctcacc cacctggata taaaggccga aaagatggaa 2220 agggagcatc taagtgttct tgaacacctg cccgccatcc gttgcctata ccttttcgtg 2280 aagcgagtct ccgaagacgg gctcgtcatc agccacggcg cgttccgatg tctacggcgt 2340 ctcgagttct gtaacgtaga tggacctggt ttgatgtttg caggaggcgt tccaatgttg 2400 gaatggctga ggctcgggtt cgacgcggat agagcgcaat cgacatacgg cggtctggag 2460 gttggcatcc agcgcctctc gtctctcaaa catgtcgtgc tcattgtatg gatggtttct 2520 gaaggcggtg atgatccagc ggagcaagcc gtctggtctg ccatcaatgg ccaagtagag 2580 atgctcccca actctccgac ggttgatatc cggtttcgta gacggagtca gctgcaggca 2640 agctcagaat aa 2652 <210> 4 <211> 883 <212> PRT <213> Zea mays <400> 4 Met Glu Asn Pro Asp Ala Gln Ala Lys Ala Trp Ala Ala Glu Met Arg 1 5 10 15 Glu Leu Ala Tyr Asp Met Glu Asp Ser Ile Asp Leu Phe Thr His His 20 25 30 Val Asp His Glu Pro Ala Asp Thr Ala Thr Thr Gly Val Lys Arg Phe 35 40 45 Phe Leu Arg Ile Ile Arg Lys Leu Lys Lys Leu His Tyr Arg His Arg 50 55 60 Phe Val Gln Glu Ile Lys Gln Leu His Asp Leu Ala Asn Glu Ser Tyr 65 70 75 80 Arg Arg Arg Lys Arg Tyr Arg Ile Glu Glu Gly Gly Ser Ser Leu Ser 85 90 95 His Ala Glu Ile Asp Pro Arg Leu Glu Ala Leu Tyr Val Glu Val Glu 100 105 110 Lys Leu Val Gly Ile Gln Gly Pro Ser Gln Glu Ile Ile Gly Gln Leu 115 120 125 Val Gly Glu Asn Ala Ala Glu Arg Arg Arg Val Val Ala Val Val Gly 130 135 140 Ser Gly Gly Ser Gly Lys Thr Thr Leu Ala Lys Gln Val Tyr Glu Lys 145 150 155 160 Ile Arg Cys Gln Phe Ser Cys Ala Ala Phe Val Ser Val Ser Gln Lys 165 170 175 Pro Asn Met Asn Ser Leu Leu Trp Glu Leu Leu Ser Gln Ile Gly Asn 180 185 190 His Gly Gly Asp Leu Gly Met Met Ala Val Gly Tyr Cys Ser Asp Lys 195 200 205 Gln Leu Ile Asp Arg Leu Arg Ser His Leu Glu Lys Gln Arg Tyr Leu 210 215 220 Val Val Ile Asp Asp Val Trp Thr Asn Ser Ala Trp Glu Thr Ile Gln 225 230 235 240 Cys Ala Leu Pro Lys Asn Ala His Ala Ser Lys Ile Ile Leu Thr Thr 245 250 255 Arg Ile Asn Ser Val Gly Gln Phe Ser Cys Thr Pro Asp Glu Gly Phe 260 265 270 Ile Tyr Gln Met Lys Pro Leu Cys Arg Asn Asp Ser Glu Asn Leu Phe 275 280 285 Leu Lys Arg Thr Leu Cys Asp Lys Asp Lys Phe Pro Ala Gln Leu Glu 290 295 300 Gly Ile Lys Asn Glu Ile Ile Glu Lys Cys Asp Gly Leu Pro Leu Ala 305 310 315 320 Ile Val Thr Leu Ala Ser Met Leu Ala Thr Lys Gln Arg Thr Arg Glu 325 330 335 Glu Trp Glu Arg Ala Leu Asp Ser Ile His Ser Met His Lys Lys Asp 340 345 350 Ser Gly Leu Glu Val Met Asp Lys Ile Leu Ser Leu Ser Tyr Arg Asp 355 360 365 Leu Pro His Asn Met Arg Asn Cys Leu Leu Tyr Leu Ser Thr Phe Pro 370 375 380 Glu Asp His Thr Ile Tyr Lys Asp Ala Leu Val Trp Arg Trp Met Ala 385 390 395 400 Glu Gly Phe Ile Ala Glu Thr Gln Gly Phe Thr Leu Glu Gln Val Ala 405 410 415 Glu Gly Tyr Phe Tyr Glu Phe Val Asn Arg Ser Leu Val Gln Pro Ile 420 425 430 Thr Leu Arg Ser Arg Tyr Glu Met Arg Gly Glu Gly Gly Cys Arg Val 435 440 445 His Asp Ile Val Leu Asn Phe Leu Ile Ser Arg Ala Ala Glu Glu Asn 450 455 460 Phe Leu Thr Thr Leu Tyr Gly Ala Gln Gly Val Pro Ser Ser Asp Arg 465 470 475 480 Arg Ile Arg Arg Leu Ser Val Trp Asp Ser Pro Glu His Ala Leu Ala 485 490 495 Val Ser Arg Ala Thr Met Asn Leu Ser His Leu Arg Ser Val Arg Ile 500 505 510 Cys Asn Val Gly Asp Trp Pro Val Pro Ala Val Leu Asp Leu Pro Val 515 520 525 Leu Arg Val Leu Asp Leu Glu Gly Cys Arg Asp Leu Arg Ile Asp Glu 530 535 540 Pro Asp Cys Ile Leu Ser Leu Phe His Leu Arg Tyr Leu Gly Phe Arg 545 550 555 560 Ser Ala Ser Gly Val Val Leu Pro Ala Gln Ile Gly Asn Leu His His 565 570 575 Leu Gln Thr Ile Asp Leu Ser Gly Thr Gly Val Thr Gln Leu Pro Glu 580 585 590 Ser Ile Val Gln Leu Lys Arg Leu Met His Leu Val Gly Gln Arg Leu 595 600 605 Ile Met Pro Asp Gly Phe Gly Ser Met Glu Ser Leu Glu Glu Leu Gly 610 615 620 Thr Ile Asp Cys Cys Lys Cys Pro Val Ser Phe Gly Glu Asp Leu Ala 625 630 635 640 Leu Leu Ser Arg Leu Arg Val Leu Arg Val Ala Phe Ile Gly Val Glu 645 650 655 Thr Ser Asp Met Glu Thr Arg Arg Lys Ser Leu Met Ser Ser Leu Cys 660 665 670 Lys Leu Gly Gly Asp Asn Leu Arg Arg Val Thr Ile Ile Asp Leu Ala 675 680 685 Gly Gly Gly Asp Cys Phe Val Glu Ser Trp His Pro Pro Pro Arg Leu 690 695 700 Leu Gln Lys Phe Ile His Ile Ser Gln His Phe Ser Arg Phe Pro Glu 705 710 715 720 Trp Ile Ser Ser Cys Leu Cys Asp Leu Thr His Leu Asp Ile Lys Ala 725 730 735 Glu Lys Met Glu Arg Glu His Leu Ser Val Leu Glu His Leu Pro Ala 740 745 750 Ile Arg Cys Leu Tyr Leu Phe Val Lys Arg Val Ser Glu Asp Gly Leu 755 760 765 Val Ile Ser His Gly Ala Phe Arg Cys Leu Arg Arg Leu Glu Phe Cys 770 775 780 Asn Val Asp Gly Pro Gly Leu Met Phe Ala Gly Gly Val Pro Met Leu 785 790 795 800 Glu Trp Leu Arg Leu Gly Phe Asp Ala Asp Arg Ala Gln Ser Thr Tyr 805 810 815 Gly Gly Leu Glu Val Gly Ile Gln Arg Leu Ser Ser Leu Lys His Val 820 825 830 Val Leu Ile Val Trp Met Val Ser Glu Gly Gly Asp Asp Pro Ala Glu 835 840 845 Gln Ala Val Trp Ser Ala Ile Asn Gly Gln Val Glu Met Leu Pro Asn 850 855 860 Ser Pro Thr Val Asp Ile Arg Phe Arg Arg Arg Ser Gln Leu Gln Ala 865 870 875 880 Ser Ser Glu <210> 5 <211> 2631 <212> DNA <213> Maize <400> 5 atggagaacc cagacgcgca ggcgaaggcg tgggcggcgg agatgcgcga gctggcctac 60 gacatggagg acagcatcga tctcttcacc caccacgtcg accacgaacc ggccgacacc 120 gccaccaccg gcgtcaagag gttcttcctc cggatcatcc ggaagcttaa gaaactccac 180 taccgccaca ggtttgctca ggagatcaaa caactccacg accttgccaa cgaatcgtac 240 cggcgtagga agaggtacag gattgaggag ggcggttcaa gcctcccgca cgcggagatc 300 gatcctcggt tagaggcgct ctacgtggag gtggagaaac tcgtgggcat ccagggccca 360 agccaggaga tcattggaca gctcgtcggc gagaacgcag cggagcggcg gagggttgtc 420 gccgttgttg gatctggagg ttcaggcaag accacacttg ccaaacaggt gtacgagaaa 480 atcaggtgcc aattctcttg tgcagccttt gtgtccgtgt cgcaaaagcc caacatgaat 540 agcctcctgt gggagttgtt atctcaaatc gggaaccatg gtggagattt aggaatgatg 600 gcagtaggat attgcagtga caaacaactg atcgacagac taagatcaca tcttgaaaag 660 cagaggtatc tcgttgtgat agatgatgtt tggacaaact cagcgtggga gaccatacaa 720 tgtgcgctcc ctaaaaatgc ccatgcaagt aaaataattc tgacaacacg aatcaacagt 780 gtaggccagt tctcctgcac tccagatgag ggttttatct atcagatgaa gcctctttgc 840 agaaacgatt ctgaaaatct gtttctgaaa aggacactat gtgataaaga taagtttcct 900 gctcagctgg aggggattaa aaacgagata atcgagaaat gcgatggttt gccactggct 960 attgttactc tagctagcat gttagctact aaacagagaa caagggaaga atgggagagg 1020 gcacttgatt caatccattc tacgcacaag aaagatagta gcctggaagt gatggacaag 1080 atactgtctc tgagttacag ggatctacct cacaacatga gaaattgctt gctgtatatc 1140 agtacatttc cagaggacca cacgatttac aaagatgctc tagtatggag atggatggct 1200 gaagggttta tcgctgaaac acaaggcttt actttggagc aggttgccga gggctacttc 1260 tacgagtttg tgaacaggag tttggttcag cccataacct tgcgttcaag atatgaaatg 1320 cgtggagaag gaggttgccg agtccatgac attgtactga acttcctcat ctctcgtgca 1380 gctgaagaga actttttaac tacgctgtat ggcgcccagg gggttccatc ttcagaccga 1440 aggattcgcc ggctctctgt ctgggacagt ccagaacacg cactggcagt ctctagagcg 1500 accatgaatc tgtcccatct ccggtcagtt agaatatgca acgttggaga ctggcccgtg 1560 cctgctgttc tagacttacc tgtccttcga gtgttagatc tagagggatg ccgtgatctg 1620 aggatcgtcg accctgactg cattctaagc ttgtttcatc tgaggtacct gggtttccgc 1680 agcgcaagtg gtgtcgtgct accggctcaa ataggaaatt tacaccatct gcagaccatc 1740 gatttaagcg ggactggagt gacacagctg ccagaaagca ttgtccagct caagcgactg 1800 atgcatcttg ttgggcaacg gctcatcatg ccagacgggt ttggtagcat ggaatccctt 1860 gaggagttag gtactatcga ctgctgcaag tgccccgctg agggtgctcc gagtgaccga 1920 gtggctttcg tcggggtcga aacaagtgac atggaaacca gaaggaaatc tttgatgtca 1980 tccctctgca aactcggagg agacaacctt cggcgtgtca ctattatcga cctcgctggc 2040 ggtggagatt gctttgtgga gtcgtggcac cctcctcctc gtctcctcca gaagttcatc 2100 catatcagtc agcaacagca cttctccagg tttccagaat ggatcagttc ctgcctatgt 2160 gatctcaccc acctggatat aaaggccgaa aagatggaaa gggagcatct aagtgttctt 2220 gaacacctgc ccgccatccg ttatctatac cttttcgtga agcgagtctc cgaagacggg 2280 ctcgtcatca gccacagcgc gttccgatgt ctacggcgtc tcgagttctg taacttagat 2340 ggacctggtt tgatgtttgc aggaggcgtt ccaatgctgg aatggctgag gctcgggttc 2400 gacgcggata gagcgcaatc gacatacggc ggtctggagg ttggcatcca gcgcctctcg 2460 tctctcaaac atgtcgtgct cattgtctgt atggtttctg aaggcggtga tgatccagcg 2520 gagcaagccg tctggtctgc catcaatggc caagtagaga tgctccccaa ttctccgacg 2580 gttgatatcc ggtttcgtag acggagtcag ctgcaggcaa gctcagaata a 2631 <210> 6 <211> 876 <212> PRT <213> Zea mays <400> 6 Met Glu Asn Pro Asp Ala Gln Ala Lys Ala Trp Ala Ala Glu Met Arg 1 5 10 15 Glu Leu Ala Tyr Asp Met Glu Asp Ser Ile Asp Leu Phe Thr His His 20 25 30 Val Asp His Glu Pro Ala Asp Thr Ala Thr Thr Gly Val Lys Arg Phe 35 40 45 Phe Leu Arg Ile Ile Arg Lys Leu Lys Lys Leu His Tyr Arg His Arg 50 55 60 Phe Ala Gln Glu Ile Lys Gln Leu His Asp Leu Ala Asn Glu Ser Tyr 65 70 75 80 Arg Arg Arg Lys Arg Tyr Arg Ile Glu Glu Gly Gly Ser Ser Leu Pro 85 90 95 His Ala Glu Ile Asp Pro Arg Leu Glu Ala Leu Tyr Val Glu Val Glu 100 105 110 Lys Leu Val Gly Ile Gln Gly Pro Ser Gln Glu Ile Ile Gly Gln Leu 115 120 125 Val Gly Glu Asn Ala Ala Glu Arg Arg Arg Val Val Ala Val Val Gly 130 135 140 Ser Gly Gly Ser Gly Lys Thr Thr Leu Ala Lys Gln Val Tyr Glu Lys 145 150 155 160 Ile Arg Cys Gln Phe Ser Cys Ala Ala Phe Val Ser Val Ser Gln Lys 165 170 175 Pro Asn Met Asn Ser Leu Leu Trp Glu Leu Leu Ser Gln Ile Gly Asn 180 185 190 His Gly Gly Asp Leu Gly Met Met Ala Val Gly Tyr Cys Ser Asp Lys 195 200 205 Gln Leu Ile Asp Arg Leu Arg Ser His Leu Glu Lys Gln Arg Tyr Leu 210 215 220 Val Val Ile Asp Asp Val Trp Thr Asn Ser Ala Trp Glu Thr Ile Gln 225 230 235 240 Cys Ala Leu Pro Lys Asn Ala His Ala Ser Lys Ile Ile Leu Thr Thr 245 250 255 Arg Ile Asn Ser Val Gly Gln Phe Ser Cys Thr Pro Asp Glu Gly Phe 260 265 270 Ile Tyr Gln Met Lys Pro Leu Cys Arg Asn Asp Ser Glu Asn Leu Phe 275 280 285 Leu Lys Arg Thr Leu Cys Asp Lys Asp Lys Phe Pro Ala Gln Leu Glu 290 295 300 Gly Ile Lys Asn Glu Ile Ile Glu Lys Cys Asp Gly Leu Pro Leu Ala 305 310 315 320 Ile Val Thr Leu Ala Ser Met Leu Ala Thr Lys Gln Arg Thr Arg Glu 325 330 335 Glu Trp Glu Arg Ala Leu Asp Ser Ile His Ser Thr His Lys Lys Asp 340 345 350 Ser Ser Leu Glu Val Met Asp Lys Ile Leu Ser Leu Ser Tyr Arg Asp 355 360 365 Leu Pro His Asn Met Arg Asn Cys Leu Leu Tyr Ile Ser Thr Phe Pro 370 375 380 Glu Asp His Thr Ile Tyr Lys Asp Ala Leu Val Trp Arg Trp Met Ala 385 390 395 400 Glu Gly Phe Ile Ala Glu Thr Gln Gly Phe Thr Leu Glu Gln Val Ala 405 410 415 Glu Gly Tyr Phe Tyr Glu Phe Val Asn Arg Ser Leu Val Gln Pro Ile 420 425 430 Thr Leu Arg Ser Arg Tyr Glu Met Arg Gly Glu Gly Gly Cys Arg Val 435 440 445 His Asp Ile Val Leu Asn Phe Leu Ile Ser Arg Ala Ala Glu Glu Asn 450 455 460 Phe Leu Thr Thr Leu Tyr Gly Ala Gln Gly Val Pro Ser Ser Asp Arg 465 470 475 480 Arg Ile Arg Arg Leu Ser Val Trp Asp Ser Pro Glu His Ala Leu Ala 485 490 495 Val Ser Arg Ala Thr Met Asn Leu Ser His Leu Arg Ser Val Arg Ile 500 505 510 Cys Asn Val Gly Asp Trp Pro Val Pro Ala Val Leu Asp Leu Pro Val 515 520 525 Leu Arg Val Leu Asp Leu Glu Gly Cys Arg Asp Leu Arg Ile Val Asp 530 535 540 Pro Asp Cys Ile Leu Ser Leu Phe His Leu Arg Tyr Leu Gly Phe Arg 545 550 555 560 Ser Ala Ser Gly Val Val Leu Pro Ala Gln Ile Gly Asn Leu His His 565 570 575 Leu Gln Thr Ile Asp Leu Ser Gly Thr Gly Val Thr Gln Leu Pro Glu 580 585 590 Ser Ile Val Gln Leu Lys Arg Leu Met His Leu Val Gly Gln Arg Leu 595 600 605 Ile Met Pro Asp Gly Phe Gly Ser Met Glu Ser Leu Glu Glu Leu Gly 610 615 620 Thr Ile Asp Cys Cys Lys Cys Pro Ala Glu Gly Ala Pro Ser Asp Arg 625 630 635 640 Val Ala Phe Val Gly Val Glu Thr Ser Asp Met Glu Thr Arg Arg Lys 645 650 655 Ser Leu Met Ser Ser Leu Cys Lys Leu Gly Gly Asp Asn Leu Arg Arg 660 665 670 Val Thr Ile Ile Asp Leu Ala Gly Gly Gly Asp Cys Phe Val Glu Ser 675 680 685 Trp His Pro Pro Pro Arg Leu Leu Gln Lys Phe Ile His Ile Ser Gln 690 695 700 Gln Gln His Phe Ser Arg Phe Pro Glu Trp Ile Ser Ser Cys Leu Cys 705 710 715 720 Asp Leu Thr His Leu Asp Ile Lys Ala Glu Lys Met Glu Arg Glu His 725 730 735 Leu Ser Val Leu Glu His Leu Pro Ala Ile Arg Tyr Leu Tyr Leu Phe 740 745 750 Val Lys Arg Val Ser Glu Asp Gly Leu Val Ile Ser His Ser Ala Phe 755 760 765 Arg Cys Leu Arg Arg Leu Glu Phe Cys Asn Leu Asp Gly Pro Gly Leu 770 775 780 Met Phe Ala Gly Gly Val Pro Met Leu Glu Trp Leu Arg Leu Gly Phe 785 790 795 800 Asp Ala Asp Arg Ala Gln Ser Thr Tyr Gly Gly Leu Glu Val Gly Ile 805 810 815 Gln Arg Leu Ser Ser Leu Lys His Val Val Leu Ile Val Cys Met Val 820 825 830 Ser Glu Gly Gly Asp Asp Pro Ala Glu Gln Ala Val Trp Ser Ala Ile 835 840 845 Asn Gly Gln Val Glu Met Leu Pro Asn Ser Pro Thr Val Asp Ile Arg 850 855 860 Phe Arg Arg Arg Ser Gln Leu Gln Ala Ser Ser Glu 865 870 875 <210> 7 <211> 2667 <212> DNA <213> Zea mays <400> 7 atggagaacc cagacgcgca ggcgaaggcg tgggcggcgg agatgcgcga gctggcctac 60 gacatggagg acagcatcga tctcttcacc caccacgtcg accacgaacc ggccgacacc 120 gccaccaccg gcgtcaagag gttcttcctc cggatcatcc ggaagcttaa gaaactccac 180 taccgccaca ggtttgctca ggagatcaaa caactccacg accttgccaa cgaatcgtac 240 cggcgtagga agaggtacag gattgaggag ggcggttcaa gcctcccgca cgcggagatc 300 It should be noted that there may be some inaccuracies in the original transcription such as "玉蜀黍" which is directly translated as "Zea mays" without considering if it's a more specific or different term in the context. Also, the "21" in ID=21 should probably be "21" for better consistency.gatcctcggt tagaggcgct ctacgtggag gtggagaaac tcgtgggcat ccagggccca 360 agccaggaga tcattggaca gctcgtcggc gagaacgcag cggagcggcg gagggttgtc 420 gccgttgttg gatctggagg ttcaggcaag accacacttg ccaaacaggt gtacgagaaa 480 atcaggtgcc aattctcttg tgcagccttt gtgtccgtgt cgcaaaagcc caacatgaat 540 agcctcctgt gggagttgtt atctcaaatc gggaaccatg gtggagattt aggaatgatg 600 gcagtaggat attgcagtga caaacaactg atcgacagac taagatcaca tcttgaaaag 660 cagagaactg atttttcaac tgcttcacaa tctgctctta ggtatctcgt tgtgatagat 720 gatgtttgga caaactcagc gtgggagacc atacaatgtg cgctccctaa aaatgcccat 780 gcaagtaaaa taattctgac aacacgaatc aacagtgtag gccagttctc ctgcactcca 840 gatgagggtt ttatctatca gatgaagcct ctttgcagaa acgattctga aaatctgttt 900 ctgaaaagga cactatgtga taaagataag tttcctgctc agctggaggg gattaaaaac 960 gagataatcg agaaatgcga tggtttgcca ctggctattg ttactctagc tagcatgtta 1020 gctactaaac agacaaag ggagaatgg gagagggcac ttgattcaat ccattctacg 1080 1140 ctacctcaca acatgagaaa ttgcttgctg tatatcaga catttccaga ggaccacacg 1200 atttacaaag atgctctagt atggagatgg atggctgaag ggtttatcgc tgaaacacaa 1260 ggctttactt tggagcaggt tgccgagggc tacttctacg agtttgtgaa caggagtttg 1320 1380 catgacattg tactgaactt cctcatctct cgtgcagctg aagagaactt tttaactacg 1440 ctgtatggcg cccaggggt tccatcttca gaccgaagga ttcgccggct ctctgtctgg 1500 gacagtccag aacacgcact ggcagtctct agagcgacca tgaatctgtc ccatctccgg 1560 tcagttagaa tatgcaacgt tggagactgg cccgtgcctg ctgttctaga cttacctgtc 1620 cttcgagtgt tagatctaga gggatgccgt gatctgagga tcgtcgaccc tgactgcatt 1680 ctaagcttgt ttcatctgag gtacctgggt ttccgcagcg caagtggtgt cgtgctaccg 1740 gctcaaatag gaaatttaca ccatctgcag accatcgatt taagcgggac tggagtgaca 1800 cagctgccag aaagcattgt ccagctcaag cgactgatgc atcttgttgg gcaacggctc 1860 atcatgccag acgggtttgg tagcatggaa tcccttgagg agttaggtac tatcgactgc 1920 tgcaagtgcc ccgctgaggg tgctccgagt gaccgagtgg ctttcgtcgg ggtcgaaaca 1980 agtgacatgg aaaccagaag gaaatctttg atgtcatccc tctgcaaact cggaggagac 2040 aaccttcggc gtgtcactat tatcgacctc gctggcggtg gagattgctt tgtggagtcg 2100 tggcaccctc ctcctcgtct cctccagaag ttcatccata tcagtcagca acagcacttc 2160 tccaggtttc cagaatggat cagttcctgc ctatgtgatc tcacccacct ggatataaag 2220 gccgaaaaga tggaaaggga gcatctaagt gttcttgaac acctgcccgc catccgttat 2280 ctataccttt tcgtgaagcg agtctccgaa gacgggctcg tcatcagcca cagcgcgttc 2340 cgatgtctac ggcgtctcga gttctgtaac ttagatggac ctggtttgat gtttgcagga 2400 ggcgttccaa tgctggaatg gctgaggctc gggttcgacg cggatagagc gcaatcgaca 2460 tacggcggtc tggaggttgg catccagcgc ctctcgtctc tcaaacatgt cgtgctcatt 2520 gtctgtatgg tttctgaagg cggtgatgat ccagcggagc aagccgtctg gtctgccatc 2580 aatggccaag tagagatgct ccccaattct ccgacggttg atatccggtt tcgtagacgg 2640 agtcagctgc aggcaagctc agaataa 2667 <210> 8 <211> 888 <212> PRT <213> Zea mays <400> 8 Met Glu Asn Pro Asp Ala Gln Ala Lys Ala Trp Ala Ala Glu Met Arg 1 5 10 15 Glu Leu Ala Tyr Asp Met Glu Asp Ser Ile Asp Leu Phe Thr His His 20 25 30 Val Asp His Glu Pro Ala Asp Thr Ala Thr Thr Gly Val Lys Arg Phe 35 40 45 Phe Leu Arg Ile Ile Arg Lys Leu Lys Lys Leu His Tyr Arg His Arg 50 55 60 Phe Ala Gln Glu Ile Lys Gln Leu His Asp Leu Ala Asn Glu Ser Tyr 65 70 75 80 Arg Arg Arg Lys Arg Tyr Arg Ile Glu Glu Gly Gly Ser Ser Leu Pro 85 90 95 His Ala Glu Ile Asp Pro Arg Leu Glu Ala Leu Tyr Val Glu Val Glu 100 105 110 Lys Leu Val Gly Ile Gln Gly Pro Ser Gln Glu Ile Ile Gly Gln Leu 115 120 125 Val Gly Glu Asn Ala Ala Glu Arg Arg Arg Val Val Ala Val Val Gly 130 135 140 Ser Gly Gly Ser Gly Lys Thr Thr Leu Ala Lys Gln Val Tyr Glu Lys 145 150 155 160 Ile Arg Cys Gln Phe Ser Cys Ala Ala Phe Val Ser Val Ser Gln Lys 165 170 175 Pro Asn Met Asn Ser Leu Leu Trp Glu Leu Leu Ser Gln Ile Gly Asn 180 185 190 His Gly Gly Asp Leu Gly Met Met Ala Val Gly Tyr Cys Ser Asp Lys 195 200 205 Gln Leu Ile Asp Arg Leu Arg Ser His Leu Glu Lys Gln Arg Thr Asp 210 215 220 Phe Ser Thr Ala Ser Gln Ser Ala Leu Arg Tyr Leu Val Val Ile Asp 225 230 235 240 Asp Val Trp Thr Asn Ser Ala Trp Glu Thr Ile Gln Cys Ala Leu Pro 245 250 255 Lys Asn Ala His Ala Ser Lys Ile Ile Leu Thr Thr Arg Ile Asn Ser 260 265 270 Val Gly Gln Phe Ser Cys Thr Pro Asp Glu Gly Phe Ile Tyr Gln Met 275 280 285 Lys Pro Leu Cys Arg Asn Asp Ser Glu Asn Leu Phe Leu Lys Arg Thr 290 295 300 Leu Cys Asp Lys Asp Lys Phe Pro Ala Gln Leu Glu Gly Ile Lys Asn 305 310 315 320 Glu Ile Ile Glu Lys Cys Asp Gly Leu Pro Leu Ala Ile Val Thr Leu 325 330 335 Ala Ser Met Leu Ala Thr Lys Gln Arg Thr Arg Glu Glu Trp Glu Arg 340 345 350 Ala Leu Asp Ser Ile His Ser Thr His Lys Lys Asp Ser Ser Leu Glu 355 360 365 Val Met Asp Lys Ile Leu Ser Leu Ser Tyr Arg Asp Leu Pro His Asn 370 375 380 Met Arg Asn Cys Leu Leu Tyr Ile Ser Thr Phe Pro Glu Asp His Thr 385 390 395 400 Ile Tyr Lys Asp Ala Leu Val Trp Arg Trp Met Ala Glu Gly Phe Ile 405 410 415 Ala Glu Thr Gln Gly Phe Thr Leu Glu Gln Val Ala Glu Gly Tyr Phe 420 425 430 Tyr Glu Phe Val Asn Arg Ser Leu Val Gln Pro Ile Thr Leu Arg Ser 435 440 445 Arg Tyr Glu Met Arg Gly Glu Gly Gly Cys Arg Val His Asp Ile Val 450 455 460 Leu Asn Phe Leu Ile Ser Arg Ala Ala Glu Glu Asn Phe Leu Thr Thr 465 470 475 480 Leu Tyr Gly Ala Gln Gly Val Pro Ser Ser Asp Arg Arg Ile Arg Arg 485 490 495 Leu Ser Val Trp Asp Ser Pro Glu His Ala Leu Ala Val Ser Arg Ala 500 505 510 Thr Met Asn Leu Ser His Leu Arg Ser Val Arg Ile Cys Asn Val Gly 515 520 525 Asp Trp Pro Val Pro Ala Val Leu Asp Leu Pro Val Leu Arg Val Leu 530 535 540 Asp Leu Glu Gly Cys Arg Asp Leu Arg Ile Val Asp Pro Asp Cys Ile 545 550 555 560 Leu Ser Leu Phe His Leu Arg Tyr Leu Gly Phe Arg Ser Ala Ser Gly 565 570 575 Val Val Leu Pro Ala Gln Ile Gly Asn Leu His His Leu Gln Thr Ile 580 585 590 Asp Leu Ser Gly Thr Gly Val Thr Gln Leu Pro Glu Ser Ile Val Gln 595 600 605 Leu Lys Arg Leu Met His Leu Val Gly Gln Arg Leu Ile Met Pro Asp 610 615 620 Gly Phe Gly Ser Met Glu Ser Leu Glu Glu Leu Gly Thr Ile Asp Cys 625 630 635 640 Cys Lys Cys Pro Ala Glu Gly Ala Pro Ser Asp Arg Val Ala Phe Val 645 650 655 Gly Val Glu Thr Ser Asp Met Glu Thr Arg Arg Lys Ser Leu Met Ser 660 665 670 Ser Leu Cys Lys Leu Gly Gly Asp Asn Leu Arg Arg Val Thr Ile Ile 675 680 685 Asp Leu Ala Gly Gly Gly Asp Cys Phe Val Glu Ser Trp His Pro Pro 690 695 700 Pro Arg Leu Leu Gln Lys Phe Ile His Ile Ser Gln Gln Gln His Phe 705 710 715 720 Ser Arg Phe Pro Glu Trp Ile Ser Ser Cys Leu Cys Asp Leu Thr His 725 730 735 Leu Asp Ile Lys Ala Glu Lys Met Glu Arg Glu His Leu Ser Val Leu 740 745 750 Glu His Leu Pro Ala Ile Arg Tyr Leu Tyr Leu Phe Val Lys Arg Val 755 760 765 Ser Glu Asp Gly Leu Val Ile Ser His Ser Ala Phe Arg Cys Leu Arg 770 775 780 Arg Leu Glu Phe Cys Asn Leu Asp Gly Pro Gly Leu Met Phe Ala Gly 785 790 795 800 Gly Val Pro Met Leu Glu Trp Leu Arg Leu Gly Phe Asp Ala Asp Arg 805 810 815 Ala Gln Ser Thr Tyr Gly Gly Leu Glu Val Gly Ile Gln Arg Leu Ser 820 825 830 Ser Leu Lys His Val Val Leu Ile Val Cys Met Val Ser Glu Gly Gly 835 840 845 Asp Asp Pro Ala Glu Gln Ala Val Trp Ser Ala Ile Asn Gly Gln Val 850 855 860 Glu Met Leu Pro Asn Ser Pro Thr Val Asp Ile Arg Phe Arg Arg Arg 865 870 875 880 Ser Gln Leu Gln Ala Ser Ser Glu 885 <210> 9 <211> 2930 <212> DNA <213> Zea mays <400> 9 atggagaacc cagacgcgca ggcgaaggcg tgggcggcgg agatgcgcga gctggcctac 60 gacatggagg acagcatcga tctcttcacc caccacgtcg accacgaacc ggccgacacc 120 gccaccaccg gcgtcaagag gttcttcctc cggatcatcc ggaagcttaa gaaactccac 180 taccgccaca ggtttgttca ggagatcaaa caactccacg accttgccaa cgaatcgtac 240 cggcgtagga agaggtacag gattgaggag ggcggttcaa gcctctcgca cgcggagatc 300 gatcctcggt tagaggcgct ctacgtggag gtggagaaac tcgtgggcat ccagggccca 360 agccaggaga tcattggaca gctcgtcggc gagaacgcag cggagcgacg gagggttgtc 420 gccgttgttg gatctggagg ttcaggcaag accacacttg ccaaacaggt gtacgagaaa 480 atcaggtgcc aattctcttg tgcagccttt gtgtctgtgt cgcaaaagcc caacatgaat 540 agcctcctgt gggagttgct atctcaaatc gggaaccatg gtggagattt aggaatgatg 600 gcagtaggat attgcagtga caaacaactg atcgacagac taagatcaca tcttgaaaag 660 cagaggttag tttacctttt cattccggtt agcttaattc ggtacaccaa ctagagattt 720 gtgattgct attaattaca ccaaatttct cctacacaac aataactggt ttagcatgat 780 ggcgatccaa agtcaaaact atcttctact actagtgtat gccatactca tatagatatt 840 ttctttcat aaactctcgt agcattttta catgcattca tattcctatt gcctttatac 900 agaactgatt ttcactgct tcacaatctg ctcttaggta tctcgttgtg atagatgatg 960 tttggacaaa ctcagcgtgg gagaccatac aatgtgcgct ccctaaaaat gcccatgcaa 1020 gtaaaataat tctgacaaca cgaatcaaca gtgtaggcca gttctcctgc actccagatg 1080 agggtttat ctatcagatg aagcctcttt gcagaaacga ttctgaaaat ctgtttctga 1140 aaaggacact atgtgataaa gataagtttc ctgctcagct ggaggggatt aaaaacgaga 1200 taatcgagaa atgcgatggt ttgccactgg ctattgttac tctagctagc atgttagcta 1260 ctaaacagag aacaagggaa gaatgggaga gggcacttga ttcaatccat tctatgcaca 1320 agaaagatag tggcctggaa gtgatggaca agatactgtc tctgagttac agggatctac 1380 ctcacaacat gagaaattgc ttgctgtatc tcagtacatt tccagaggac cacacgattt 1440 acaaagatgc cctagtatgg agatggatgg ctgaagggtt tatcgctgaa acacaaggct 1500 ttactttgga gcaggttgcc gagggctact tctacgagtt tgtgaacagg agtttggttc 1560 agcccataac cttgcgttca agatatgaaa tgcgtggaga aggaggttgc cgagtccatg 1620 acattgtact gaacttcctc atctctcgtg cagctgaaga gaacttttta actacgctgt 1680 atggcgccca gggggttcca tcttcagacc gaaggattcg ccggctctct gtctgggaca 1740 gtccagaaca cgcactggca gtctctagag cgaccatgaa tctgtcccat ctccggtcag 1800 ttagaatatg caacgttgga gactggcccg tgcctgctgt tctagactta cctgtccttc 1860 gagtgttaga tctagaggga tgccgtgatc tgaggatcga cgaacctgac tgcattctaa 1920 gcttgttca tctgagatac ctgggtttcc gcagcgcaag tggtgtcgtg ctaccggctc 1980 aaatcggaaa tttacaccat ctgcagacca tcgatttaag cgggactgga gtgacacagc 2040 tgccagaaag cattgtccag ctcaagcgac tgatgcatct tgttgggcaa cggctcatca 2100 tgccagacgg gtttggtagc atggaatccc ttgaggagtt aggtactatc gactgctgca 2160 agtgccccgt cagttttggg gaagacctag cacttctgag caggctgagg gtgctccgag 2220 tggctttcat cggggtcgaa acaagtgaca tggaaaccag aaggaaatct ttgatgtcat 2280 ccctctgcaa actcggagga gacaaccttc ggcgtgtcac tattatcgac ctcgctggcg 2340 gtggagattg cttgtggag tcgtggcacc ctcctcctcg tctcctccag aagttcatcc 2400 atatcagtca gcaacagcac ttctccaggt ttccagaatg gatcagttcc tgcctatgtg 2460 atctcaccca cctggatata aaggccgaaa agatggaaag ggagcatcta agtgttcttg 2520 aacacctgcc cgccatccgt tgcctatacc tttcgtgaa gcgagtctcc gaagacgggc 2580 tcgccatcag ccacggcgcg ttccgatgtc tacggcgtct cgagttctgc aacgtagatg 2640 gacctggtt gatgtttgca ggaggcgttc caatgttgga atggctgagg ctcgggttcg 2700 acgcggatag agcgcaatcg acatacggcg gtctggaggt tggcatccag cgcctctcgt 2760 ctctcaaaca tgtcgtgctc attgtatgga tggtttctga aggcggtgat gatccagcgg 2820 agcaagccgt ctggtctgcc atcaatggcc aagtagagat gctccccaac tctccgacgg 2880 ttgatatccg gtttcgtaga cggagtcagc tgcaggcaag ctcagaataa 2930 <210> 10 <211> 16 <212> PRT <213> Zea mays <400> 10 Val Ser Phe Gly Glu Asp Leu Ala Leu Leu Ser Arg Leu Arg Val Leu 1 5 10 15 <210> 11 <211> 667 <212> PRT <213> Zea mays <400> 11 Met Ala Ala His Gln Pro His Leu Ser Val Leu Leu Leu Val Leu Leu 1 5 10 15 Ala Ala His Val Val Ser Thr Ser Ala His Gly Glu Pro Pro Leu Pro 20 25 30 Ser Pro Tyr Asn Thr Ser Ala His Gly Glu Pro Pro Leu Pro Ser Thr 35 40 45 Tyr Asn Ala Ser Met Cys Ser Ser Phe Trp Cys Gly Gly Val Glu Ile 50 55 60 Arg Tyr Pro Phe Tyr Leu Ala Asn Ala Ile Ala Asp Tyr Ser Gly Ser 65 70 75 80 Tyr Tyr Ser Cys Gly Tyr Thr Asp Leu Ser Val Ser Cys Glu Leu Glu 85 90 95 Val Glu Gly Ser Pro Thr Thr Trp Thr Pro Thr Ile Arg Leu Gly Gly 100 105 110 Gly Asp Tyr Thr Val Lys Asn Ile Ser Tyr Leu Tyr Asp Gln Gln Thr 115 120 125 Ile Ser Leu Ala Asp Arg Asp Val Leu Gly Gly Gly Gly Cys Pro Val 130 135 140 Val Arg His Asn Val Ser Phe Asp Glu Thr Trp Leu His Leu His Asn 145 150 155 160 Ala Ser Ala Phe Asp Asn Leu Thr Phe Phe Phe Gly Cys His Trp Gly 165 170 175 Pro Arg Asn Thr Pro Pro Glu Phe Ala Asp Tyr Asn Ile Ser Cys Ala 180 185 190 Gly Phe Asn Thr Pro Thr Ile Ser Gly Gly Arg Ser Phe Val Phe Lys 195 200 205 Thr Gly Asp Leu Asp Glu Gln Glu Glu Gln Glu Leu Ala Leu His Cys 210 215 220 Asp Glu Val Phe Ser Val Pro Val Arg Arg Asp Ala Leu Gln Ala Ile 225 230 235 240 Val Ser Asn Phe Ser Leu Thr Arg Asp Gly Tyr Gly Glu Val Leu Arg 245 250 255 Gln Gly Phe Glu Leu Glu Trp Asn Arg Thr Ser Glu Asp Gln Cys Gly 260 265 270 Arg Cys Glu Gly Ser Gly Ser Gly Gly Trp Cys Ala Tyr Ser Gln Lys 275 280 285 Arg Glu Phe Leu Gly Cys Leu Cys Ser Gly Gly Lys Val Gly Ser Pro 290 295 300 Phe Cys Lys Pro Ser Arg Ser Lys Arg Lys Glu Gly Pro Ile Val Gly 305 310 315 320 Ala Val Ala Val Ala Phe Leu Cys Leu Val Ile Leu Thr Cys Phe Leu 325 330 335 Ala Cys Arg His Gly Ser Leu Pro Phe Lys Ser Glu Asn Lys Pro Gly 340 345 350 Thr Arg Ile Glu Ser Phe Leu Gln Lys Asn Glu Ser Ile His Pro Lys 355 360 365 Arg Tyr Thr Tyr Ala Asp Val Lys Arg Met Thr Lys Ser Phe Ala Val 370 375 380 Lys Leu Gly Gln Gly Gly Phe Gly Ala Val Tyr Lys Gly Ser Leu His 385 390 395 400 Asp Gly Arg Gln Val Ala Val Lys Met Leu Lys Asp Thr Gln Gly Asp 405 410 415 Gly Glu Glu Phe Met Asn Glu Val Ala Ser Ile Ser Arg Thr Ser His 420 425 430 Val Asn Val Val Thr Leu Leu Gly Phe Cys Leu Gln Gly Ser Lys Arg 435 440 445 Ala Leu Ile Tyr Glu Tyr Met Pro Asn Gly Ser Leu Glu Arg Tyr Ala 450 455 460 Phe Thr Gly Asp Met Asn Ser Glu Asn Leu Leu Thr Trp Glu Arg Leu 465 470 475 480 Phe Asp Ile Ala Ile Gly Thr Ala Arg Gly Leu Glu Tyr Leu His Arg 485 490 495 Gly Cys Asn Thr Arg Ile Val His Phe Asp Ile Lys Pro His Asn Ile 500 505 510 Leu Leu Asp Gln Asp Phe Cys Pro Lys Ile Ser Asp Phe Gly Leu Ala 515 520 525 Lys Leu Cys Leu Asn Lys Glu Ser Ala Ile Ser Ile Ala Gly Ala Arg 530 535 540 Gly Thr Ile Gly Tyr Ile Ala Pro Glu Val Tyr Ser Lys Gln Phe Gly 545 550 555 560 Ile Ile Ser Ser Lys Ser Asp Val Tyr Ser Tyr Gly Met Met Val Leu 565 570 575 Glu Met Val Gly Ala Arg Asp Arg Asn Thr Ser Ala Asp Ser Asp His 580 585 590 Ser Ser Gln Tyr Phe Pro Gln Trp Leu Tyr Glu His Leu Asp Asp Tyr 595 600 605 Cys Val Gly Ala Ser Glu Ile Asn Gly Glu Thr Thr Glu Leu Val Arg 610 615 620 Lys Met Ile Val Val Gly Leu Trp Cys Ile Gln Val Ile Pro Thr Asp 625 630 635 640 Arg Pro Thr Met Thr Arg Val Val Glu Met Leu Glu Gly Ser Thr Ser 645 650 655 Asn Leu Glu Leu Pro Pro Arg Val Leu Leu Ser 660 665 <210> 12 <211> 666 <212> PRT <213> Maize <400> 12 Met Ala Ala His Leu Pro Arg Leu Pro Val Leu Leu Leu Val Leu Leu 1 5 10 15 Ala Ala His Val Val Ser Thr Ser Ala His Ala Glu Pro Pro Leu Pro 20 25 30 Ser Pro Tyr Ser Thr Ser Ala His Gly Glu Pro Pro Leu Pro Ser Thr 35 40 45 Tyr Asn Val Ser Met Cys Ser Glu Ser Phe Trp Cys Gly Gly Val Glu 50 55 60 Ile Arg Tyr Pro Phe Tyr Leu Ala Asn Ala Thr Ala Asp Tyr Ser Gly 65 70 75 80 Ser Tyr Tyr Ser Cys Gly Tyr Thr Asp Leu Ser Val Ser Cys Lys Leu 85 90 95 Glu Val Glu Gly Pro Thr Thr Thr Trp Thr Pro Thr Ile Arg Leu Gly 100 105 110 Gly Asp Asn Tyr Thr Val Lys Asn Ile Leu Tyr Asp Tyr His Thr Ile 115 120 125 Ser Leu Ala Asp Ser Asp Val Leu Gly Gly Gly Glu Cys Pro Val Val 130 135 140 His His Asn Val Ser Phe Asp Glu Thr Trp Leu His Asn Pro Ser Ala 145 150 155 160 Phe Asp Asn Leu Thr Phe Phe Phe Gly Cys His Trp Gly Pro Arg Asp 165 170 175 Thr Leu Pro Glu Phe Ala Gly Asn Asn Ile Ser Cys Ala Gly Phe Ser 180 185 190 Thr Pro Ala Ile Ser Gly Gly Gly Ser Phe Val Phe Lys Pro Glu Asp 195 200 205 Leu Asp Glu His Ala Glu Gln Glu Leu Ala Ser His Cys Asp Glu Val 210 215 220 Phe Ser Val Pro Val Arg Ser Glu Ala Leu Gln Gln Ala Ile Val Ser 225 230 235 240 Asn Leu Ser Leu Gly Asp Gly Tyr Gly Glu Leu Leu Arg Gln Gly Ile 245 250 255 Glu Leu Glu Trp Lys Arg Thr Ser Glu Asp Gln Cys Gly Gln Cys Glu 260 265 270 Glu Ser Gly Ser Gly Gly Arg Cys Ala Tyr Ser Gln Lys Arg Glu Phe 275 280 285 Leu Gly Cys Leu Cys Ser Gly Gly Lys Ala Gly Asn Pro Phe Cys Lys 290 295 300 Pro Ser Arg Ser Lys Arg Lys Glu Ala Ser Ile Val Gly Ala Val Ala 305 310 315 320 Val Ala Phe Leu Cys Leu Val Ile Leu Thr Cys Phe Leu Ala Cys Arg 325 330 335 His Gly Ser Leu Pro Phe Lys Ser Glu Asn Lys Pro Gly Thr Arg Ile 340 345 350 Glu Ser Phe Leu Gln Lys Asn Glu Ser Ile His Pro Lys Arg Tyr Thr 355 360 365 Tyr Thr Asp Val Lys Arg Met Thr Lys Ser Phe Ala Val Lys Leu Gly 370 375 380 Gln Gly Gly Phe Gly Ala Val Tyr Lys Gly Ser Leu His Asp Gly Arg 385 390 395 400 Gln Val Ala Val Lys Met Leu Lys Asp Thr Gln Gly Asp Gly Glu Glu 405 410 415 Phe Met Asn Glu Val Ala Ser Ile Ser Arg Thr Ser His Val Asn Val 420 425 430 Val Thr Leu Leu Gly Phe Cys Leu Gln Gly Ser Lys Arg Ala Leu Ile 435 440 445 Tyr Glu Tyr Met Pro Asn Gly Ser Leu Glu Arg Tyr Ala Phe Thr Gly 450 455 460 Asp Met Asn Ser Glu Asn Leu Leu Thr Trp Glu Arg Leu Phe Asp Ile 465 470 475 480 Ala Ile Gly Thr Ala Arg Gly Leu Glu Tyr Leu His Arg Gly Cys Asn 485 490 495 Thr Arg Ile Val His Phe Asp Ile Lys Pro His Asn Ile Leu Leu Asp 500 505 510 Gln Asp Phe Cys Pro Lys Ile Ser Asp Phe Gly Leu Ala Lys Leu Cys 515 520 525 Leu Asn Lys Glu Ser Ala Ile Ser Ile Val Gly Ala Arg Gly Thr Ile 530 535 540 Gly Tyr Ile Ala Pro Glu Val Tyr Ser Lys Gln Phe Gly Thr Ile Ser 545 550 555 560 Ser Lys Ser Asp Val Tyr Ser Tyr Gly Met Met Val Leu Glu Met Val 565 570 575 Gly Ala Arg Glu Arg Asn Thr Ser Ala Ser Ala Asp Ser Asp His Ser 580 585 590 Ser Gln Tyr Phe Pro Gln Trp Ile Tyr Glu His Leu Asp Asp Tyr Cys 595 600 605 Val Gly Ala Ser Glu Ile Asn Gly Glu Thr Thr Glu Leu Val Arg Lys 610 615 620 Met Ile Val Val Gly Leu Trp Cys Ile Gln Val Ile Pro Thr Asp Arg 625 630 635 640 Pro Thr Met Thr Arg Val Val Glu Met Leu Glu Gly Ser Thr Ser Asn 645 650 655 Leu Glu Leu Pro Pro Arg Val Leu Leu Ser 660 665

Claims

1. A polynucleotide construct comprising isolated polynucleotides selected from the following: nucleotide sequences listed in SEQ ID NO: 1 (PH4GP c-DNA), SEQ ID NO: 3 (PH1W2 cDNA), and SEQ ID NO: 9 (PH4GP genomic sequence). The isolated polynucleotide is operatively linked to a promoter, or, in the case of SEQ ID NO: 9, contains a promoter.

2. The polynucleotide construct of claim 1, wherein the polynucleotide construct further comprises one or more additional heteronucleotide sequences encoding a polypeptide selected from: a polypeptide conferring disease resistance, a polypeptide conferring herbicide resistance, a polypeptide conferring insect resistance, a polypeptide involved in carbohydrate metabolism, a polypeptide involved in fatty acid metabolism, a polypeptide involved in amino acid metabolism, a polypeptide involved in plant development, a polypeptide involved in plant growth regulation, a polypeptide involved in yield enhancement, a polypeptide involved in drought resistance, a polypeptide involved in cold resistance, a polypeptide involved in heat resistance, and / or a polypeptide involved in salt resistance, wherein each heteronucleotide sequence is operatively linked to a promoter.

3. The polynucleotide construct of claim 2, wherein the disease-conferring polypeptide is a polypeptide that confers resistance to Northern Leaf Blight (NLB).

4. The polynucleotide construct of claim 3, wherein the polypeptide conferring resistance to northern leaf blight is a polypeptide consisting of the amino acid sequence listed in SEQ ID NO: 11 or 12.

5. A method for producing corn plant cells, said corn plant cells comprising the polynucleotide construct of claim 1.

6. A method for producing a corn plant, said corn plant comprising the corn plant cells as described in claim 5.

7. A method for producing corn plants resistant to Northern Leaf Blight (NLB), the method comprising: a) Expressing a heterologous polynucleotide construct in regenerable corn plant cells, the polynucleotide construct comprising a polynucleotide selected from the nucleotide sequences listed in SEQ ID NO: 1 (PH4GP c-DNA), SEQ ID NO: 3 (PH1W2 cDNA), and SEQ ID NO: 9 (PH4GP genomic sequence), wherein the polynucleotide is operatively linked to at least one regulatory sequence, or, in the case of SEQ ID NO: 9, comprises at least one regulatory sequence; and b) Produce corn plants that exhibit resistance to northern leaf blight, wherein the corn plants contain the heteropolynucleotide construct in their genome.

8. The method of claim 7, wherein the at least one regulatory sequence is a promoter.

9. The method of claim 7, wherein the at least one regulation sequence is a terminator.

10. The method of claim 7, wherein the regulatory sequence is natural for corn.

11. The method of claim 7, wherein the regulatory sequence is natural for the Ht1 gene.

12. The method of claim 7, wherein the polynucleotide construct comprises one or more additional heteronucleotide sequences encoding a polypeptide selected from: a polypeptide conferring disease resistance, a polypeptide conferring herbicide resistance, a polypeptide conferring insect resistance, a polypeptide involved in carbohydrate metabolism, a polypeptide involved in fatty acid metabolism, a polypeptide involved in amino acid metabolism, a polypeptide involved in plant development, a polypeptide involved in plant growth regulation, a polypeptide involved in yield enhancement, a polypeptide involved in drought resistance, a polypeptide involved in cold resistance, a polypeptide involved in heat resistance, and / or a polypeptide involved in salt resistance, wherein each heteronucleotide sequence is operatively linked to a promoter.

13. The method of claim 12, wherein the disease-conferring polypeptide is a polypeptide that confers resistance to Northern Leaf Blight (NLB).

14. The method of claim 13, wherein the polypeptide conferring resistance to northern leaf blight is a polypeptide consisting of the amino acid sequence listed in SEQ ID NO: 11 or 12.

15. A method for obtaining corn plants resistant to Northern Leaf Blight (NLB), the method comprising: a) Hybridize the corn plants produced by the method of claim 7 with corn plants whose genomes do not contain the polynucleotide construct; b) Obtain progeny plants that exhibit resistance to northern leaf blight, wherein the progeny plants contain the polynucleotide construct in their genome.

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