Methods for identifying, selecting and producing disease-resistant crops

Abstract

This field relates to plant breeding and methods for identifying, selecting, and producing disease-resistant crops. Methods and uses of identifying novel genes encoding proteins that confer plant disease resistance are provided. These disease resistance genes can be used to produce resistant plants through breeding, transgenic modification, or genome editing.

Description

[0001] References to electronically submitted sequence listings

[0002] The sequence listing submitted with this specification in computer readable form is named "RTS22658A_SeqList.txt", created on October 15, 2020, and is 114 kilobytes in size. The sequence listing is part of this specification and is incorporated herein by reference in its entirety. Technical Field

[0003] This field relates to plant breeding and methods for identifying and selecting plants with disease resistance. Methods and uses thereof are provided for identifying novel genes encoding proteins that confer plant disease resistance. These disease resistance genes can be used to generate resistant plants through breeding, transgenic modification, or genome editing. Background Art

[0004] Southern corn rust (SCR) is a fungal disease caused by Puccinia polysora Underw that is a major disease in tropical regions and southern regions of the United States and China. If SCR reaches a critical point in temperate regions (e.g., the Midwestern United States) during the growing season, and if conditions are favorable for the occurrence of rust, disease intensity can quickly reach epidemic levels, resulting in severe yield losses. Temperate corn germplasm is generally susceptible to SCR. The identification and utilization of resistant lines and QTLs in breeding programs to develop varieties resistant to SCR represents a cost-effective way to control SCR. Alternatively, varieties carrying genes responsible for SCR resistance can be developed through transgenic or genome editing technologies. Identification of resistance QTLs and genes will accelerate the development of products resistant to SCR.Resistant lines (e.g., Brewbaker, JL, et al. "General resistance in maize to southern rust (Puccinia polysora Underw." Crop science 51, no. 4 (2011): 1393-1409) or QTLs (e.g., Jines, MP, et al. "Mapping resistance to Southern rust in a tropical by temperate maize recombinant inbred topcross population." Theoretical and Applied Genetics 114, no. 4 (2007): 659-667. Zhang, Y., et al. "Mapping of southern corn rust-resistant genes in the W2D inbred line of maize (Zea mays L.). "Positioning of a southern corn rust resistance gene in a W2D maize inbred line (maize)." Molecular breeding 25, no. 3 (2010): 433-439. Zhou CJ, et al. (2007) Characterization and fine mapping of RppQ, a resistance gene to southern corn rust in maize. Mol Genet Genomics 278: 723-728. Holland, JB, et al. "Inheritance of resistance to southern corn rust in tropical-by-corn-belt maize populations." Theoretical and Applied Genetics [Theoretical and Applied Genetics] 96, No. 2 (1998): 232-241.). However, the causal gene responsible for SCR resistance has not yet been identified and characterized. There is a continuing need for disease-resistant plants and methods for finding disease-resistance genes. Summary of the Invention

[0005] Provided herein are compositions and methods useful in identifying and selecting plant disease resistance genes, or "R genes." The compositions and methods can be used to select disease-resistant plants, generate transgenic resistant plants, and / or generate plants with genome-edited resistance genes. Also provided herein are plants having newly conferred or enhanced resistance to various plant diseases compared to control plants. In some embodiments, the compositions and methods can be used to select disease-resistant corn plants, including southern corn rust (SCR) disease-resistant plants, generate transgenic disease-resistant plants, and / or generate plants with genome-edited disease resistance genes.

[0006] The disease-resistant plant can be crossed with a second plant to obtain a progeny plant having the resistance gene allele. The disease resistance can be newly conferred or enhanced relative to a control plant that does not have the favorable allele. The R gene allele can be further refined to a chromosomal interval defined by and including a defined marker. In some embodiments, methods for identifying and / or selecting plants with disease resistance are presented. In these methods, the DNA of the plant is analyzed for the presence of a resistance gene allele associated with disease resistance on chromosome 7, wherein the resistance gene allele comprises a sequence that is at least 95% identical to SEQ ID NOs: 1-10, or 13-16; and if the resistance gene allele is detected, the plant is identified and / or selected as having disease resistance. In some embodiments, the method for identifying and / or selecting plants with disease resistance comprises detecting or selecting a genomic region comprising any one of SEQ ID NOs: 4-6. The disease resistance can be newly conferred or enhanced relative to a control plant that does not have the favorable allele. In another embodiment, the disease resistance region comprises a gene encoding a ZmMM1 polypeptide that confers or enhances disease resistance (a "ZmMM1" gene). In some embodiments, the ZmMM1 polypeptide comprises an amino acid sequence as set forth in any one of SEQ ID NOs: 1-3.

[0007] In another embodiment, a method for identifying and / or selecting plants with disease resistance is provided, wherein one or more marker alleles linked to and associated with any one of SEQ ID NOs: 1-10 or 13-16 are detected in a plant, and plants having the one or more marker alleles are selected. The one or more marker alleles can be linked at 10 cM, 9 cM, 8 cM, 7 cM, 6 cM, 5 cM, 4 cM, 3 cM, 2 cM, 1 cM, 0.9 cM, 0.8 cM, 0.7 cM, 0.6 cM, 0.5 cM, 0.4 cM, 0.3 cM, 0.2 cM or 0.1 cM or less on a genetic map based on a single meiosis. The selected plant can be crossed with a second plant to obtain a progeny plant having the one or more marker alleles linked to and associated with any one of SEQ ID NOs: 1-10 or 13-16.

[0008] In another embodiment, methods for introgressing a gene allele associated with disease resistance are provided herein. In these methods, a population of plants is screened using one or more markers to determine whether any of the plants possess the gene allele associated with disease resistance, and at least one plant from the population having the gene allele associated with disease resistance is selected. The allele comprises a sequence that is at least 95% identical to SEQ ID NOs: 1-10 or 13-16.

[0009] In some embodiments, introgression of disease resistance genes from resistant lines into susceptible lines can be achieved through marker-assisted trait introgression, transgenics, or genome editing approaches.

[0010] Embodiments include isolated polynucleotides comprising a nucleotide sequence encoding a ZmMM1 polypeptide capable of conferring disease resistance, wherein the ZmMM1 polypeptide has an amino acid sequence that is at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to any one of SEQ ID NOs: 1-3. In another embodiment, an isolated polynucleotide comprises a nucleotide sequence encoding a ZmMM1 polypeptide capable of conferring resistance, wherein the ZmMM1 polypeptide has an amino acid sequence that is at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, and at least 95% identical to any one of SEQ ID NOs: 1-3.

[0011] Additional embodiments of the present disclosure include vectors comprising a polynucleotide of the present disclosure, such as SEQ ID NOs: 4-10, or recombinant DNA constructs comprising a polynucleotide disclosed herein operably linked to at least one regulatory sequence. Plant cells and plants, each comprising a recombinant DNA construct of the embodiments disclosed herein, and seeds comprising the recombinant DNA construct are also presented.

[0012] In some embodiments, the compositions and methods relate to modified plants with increased resistance to disease, wherein the allele that causes increased disease resistance comprises a nucleotide sequence encoding a ZmMM1 resistance gene, wherein the ZmMM1 resistance gene is at least 50%, at least 75%, at least 80%, at least 85%, at least 90%, and at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the sequence shown in SEQ ID NOs: 4-10. In some embodiments, downregulation of ZmMT1 (SEQ ID NOs: 20 and 21), ZmMT2 (SEQ ID NO: 23), ZmMT3, or ZmMT4 (SEQ ID NO: 25) in maize provides enhanced resistance. Downregulation can be induced by editing or transgenic means (including RNAi knockout).

[0013] The methods embodied in the present disclosure relate to methods for transforming host cells, including plant cells, comprising transforming the host cells with a polynucleotide of an embodiment of the present disclosure; for making a plant cell comprising transforming the plant cell with a recombinant DNA construct of an embodiment of the present disclosure and regenerating a plant from the transformed plant cell, and for conferring or enhancing disease resistance, comprising transforming a plant with a recombinant DNA construct disclosed herein.

[0014] Also presented are methods for altering the expression level of a protein capable of conferring disease resistance on a plant or plant cell, the methods comprising (a) transforming a plant cell with a recombinant DNA construct disclosed herein, and (b) culturing the transformed plant cell under conditions suitable for expression of the recombinant DNA construct, wherein expression of the recombinant DNA construct results in the production of altered levels of the protein capable of conferring disease resistance in the transformed host.

[0015] Also provided are plants identified and / or selected using any of the above methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1Shown is a 5 kb interval finely mapped to 20 SNPs and 7 indels on chromosome 7 flanked by markers M2 (SEQ ID NOs: 27 and 28) and M3 (SEQ ID NOs: 11 and 12) and located between C117 and Mo17 within the 1 kb qLMchr7 region.

[0017] Sequence Description

[0018]

[0019] DETAILED DESCRIPTION

[0020] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells, and reference to "a protein" includes reference to one or more proteins and their equivalents, and so forth. All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs, unless expressly indicated otherwise.

[0021] The NBS-LRR ("NLR") group of R genes is the largest class of R genes discovered to date. In Arabidopsis thaliana, over 150 NLR genes are predicted to be present in the genome (Meyers et al., (2003), Plant Cell, 15:809-834; Monosi et al., (2004), Theoretical and Applied Genetics, 109:1434-1447), while in rice, approximately 500 NLR genes have been predicted (Monosi, (2004) supra). The NBS-LRR class of R genes consists of two subclasses. Class 1 NLR genes contain a TIR-Toll / interleukin-1-like domain at their N' terminus; to date, they have only been found in dicots (Meyers, (2003) supra; Monosi, (2004) supra). The second type of NBS-LRR contains a coiled-coil domain or (nt) domain at its N-terminus (Bai et al. (2002) Genome Research, 12:1871-1884; Monosi, (2004) supra; Pan et al. (2000) Journal of Molecular Evolution, 50:203-213). Type 2 NBS-LRRs are found in both dicot and monocot species (Bai, (2002) supra; Meyers, (2003) supra; Monosi, (2004) supra; Pan, (2000) supra).

[0022] The NBS domain of this gene appears to play a role in signaling of plant defense mechanisms (van der Biezen et al., (1998), Current Biology: CB, 8: R226-R227). The LRR region appears to be a region that interacts with pathogen AVR products (Michelmore et al., (1998), Genome Res., 8: 1113-1130; Meyers, (2003) supra). Compared to the NB-ARC (NBS) domain, this LRR region is under greater selective pressure to diversify (Michelmore, (1998) supra; Meyers, (2003) supra; Palomino et al., (2002), Genome Research, 12: 1305-1315). LRR domains can also be found in other contexts; these 20-29 residue motifs are arranged in tandem in many proteins that have diverse functions, such as hormone-receptor interactions, enzyme inhibition, cell adhesion, and cell trafficking. Many recent studies have shown that LRR proteins are involved in early mammalian development, neural development, cell polarization, regulation of gene expression, and apoptosis signaling.

[0023] An allele is "associated with a trait" when it is part of or linked to a DNA sequence or allele that affects the expression of the trait. The presence of the allele is an indicator of how the trait will be expressed.

[0024] As used herein, "disease resistance" or "resistant to a disease" refers to a plant that exhibits increased resistance to a disease compared to a control plant. Disease resistance can be manifested as fewer and / or smaller lesions, increased plant health, increased yield, increased root mass, increased plant vigor, less or no discoloration, increased growth, reduced necrotic area, or reduced wilting. In some embodiments, the allele can confer resistance to one or more diseases.

[0025] Diseases affecting corn plants include, but are not limited to, bacterial leaf blight and stalk rot; bacterial leaf spot; bacterial stripe; chocolate spot; goss's bacterial wilt and blight; holcus spot; purple leaf sheath; seed rot-seedling blight; bacterial wilt; corn stunt; anthracnose leaf blight; anthracnose stalk rot; aspergillus ear and kernel rot; banded leaf and sheath spot; and corn stunt. leaf and sheath spot; black bundle disease; black kernel rot; borde blanco; brown spot; blackspot; stalk rot; cephalosporium kernel rot; charcoal rot; corticium ear rot; curvularia leaf spot; didymella leaf spot; diplodia ear rot and stalk rot; diplodia ear rot; seed rot; corn seedling blight; diplodia leaf spot or leafstreak; downy mildew mildews); brown stripe downy mildew; crazy top downy mildew; green ear downy mildew; graminicola downy mildew;Java downy mildew; Philippine downy mildew; sorghum downy mildew; spontaneum downy mildew; sugarcane downy mildew; dry earrot; ergot; horse's tooth; corn eyespot; fusarium ear and stalk rot; fusarium blight; seedling root rot; gibberella ear and stalk rot; gray ear rot; gray leaf spot; cercospora leaf spot; helminthosporium root rot rot; hormodendrum ear rot; cladosporium rot; hyalothyridium leaf spot; late wilt; northern leaf blight; white blast; crown stalk rot; corn stripe; northern leaf spot; helminthosporium ear rot; penicillium ear rot; corn blue eye; blue mold; phaeocytostromastalk rot and root rot; phaeosphaeria leaf spot; physalospora ear rot rot); botryosphaeria ear rot; pyrenochaeta stalk rot and root rot; pythium root rot; pythium stalk rot;Red kernel disease; Rhizoctonia ear rot; Sclerotial rot; Rhizoctonia root rot and stalk rot; Rostratum leaf spot; Common corn rust; Southern corn rust; Tropical corn rust; Sclerotium ear rot; Southern blight; Selenophoma leaf spot; Sheath rot; Shuck rot; Silage mold; Common smut; False smut; Head smut; Southern corn leaf blight and stalk rot rot; southern leaf spot; tar spot; trichoderma ear rot and root rot; white ear rot, root and stalk rot; yellow leaf blight; zonate leaf spot; wheat striate mosaic; barley stripe mosaic; barley yellow dwarf; bromemosaic; cereal chlorotic mottle; maize lethal necrosis disease; cucumber mosaic; Johnsongrass mosaic virus; maize bushy dwarf stunt); maize chlorotic dwarf; maize chlorotic mottle; maize dwarf mosaic;Maize leaffleck; maize pellucid ringspot; maize rayado fino; maize red leaf and red stripe; maize red stripe; maize ring mottle; maize rough dwarf; maize sterile stunt; maize streak; maize stripe; maize tassel abortion; maize vein enation; maize wallaby ear); maize white leaf; maize white line mosaic; millet red leaf; and northern cereal mosaic.

[0026] Diseases affecting plants include, but are not limited to, bacterial blight; bacterial leaf streak; foot rot; grain rot; sheath brown rot; blast; brown spot; crown sheath rot; downy mildew; eyespot; false smut; kernel smut; leaf smut; leaf scald; narrow brown leaf spot; root rot; seedling blight; sheath blight; sheath rot; sheath spot; alternaria leaf spot; spot); and stem rot.

[0027] Diseases affecting soybean plants include, but are not limited to, alternaria leaf spot; anthracnose; black leaf blight; black root rot; brown spot; brown stem rot; charcoal rot; choanephora leaf blight; downy mildew; drechslerablight; frogeye leaf spot; leptosphaerulina leaf spot; mycoleptodiscus root rot; neocosmospora stem rot; phomopsis seed decay; phytophthora root and stem rot. rot; phyllosticta leaf spot; phymatotrichum root rot; pod and stem blight; powdery mildew; purple seed stain; pyrenochaeta leaf spot; pythium rot; red crown rot; dactuliophora leaf spot; rhizoctonia aerial blight; rhizoctonia root and stem rot; rust; scab; sclerotinia stem rot; sclerotium blight; stem canker; stemphylium leafblight; sudden death syndrome; target spot; yeast spot; lance nematode; lesion nematode; pin nematode;reniform nematode; ring nematode; root-knot nematode; sheath nematode; cyst nematode; spiral nematode; sting nematode; stubby root nematode; stunt nematode; alfalfa mosaic; bean pod mottle; bean yellow mosaic; Brazilian bud blight; chlorotic mottle; yellow mosaic; peanut mottle; peanut stripe; peanut stunt; chlorotic mottle mottle; crinkle leaf; dwarf; severe stunt; and tobacco ringspot or bud blight.

[0028] Diseases affecting canola plants include, but are not limited to, bacterial blackrot; bacterial leaf spot; bacterial pod rot; bacterial soft rot; scab; crown gall; alternaria black spot; anthracnose; black leg; black mold rot; black root; brown girdling root rot; cercospora leaf spot; clubroot; downy mildew; fusarium wilt; gray mold; head rot; leaf spot; light leaf spot; and spot; pod rot; powdery mildew; ringspot; root rot; sclerotinia stem rot; seedrot (damping-off); root gall smut; southern blight; verticillium wilt; white blight; white leaf spot; staghead; yellows; crinkle virus; mosaic virus; yellows virus;

[0029] Diseases affecting sunflower plants include, but are not limited to, apical chlorosis; bacterial leaf spot; bacterial wilt; crown gall; erwinia stalk rot and head rot; alternaria leaf blight, stem spot and head rot; botrytis headrot; charcoal rot; downy mildew; fusarium stalk rot; fusarium wilt; myrothecium leaf and stem spot; phialophora yellows; phoma blackstem; phomopsis brown stem canker. canker; phymatotrichum root rot; phytophthora stem rot; powdery mildew; pythium seedling blight and root rot; rhizoctonia seedling blight; rhizopus head rot; sunflower rust; sclerotium basal stalk and root rot; septorial eaf spot; verticillium wilt; white rust; yellow stripe rust; scimitar; needle-shaped; lesions; reniform; knots; and chlorotic mottle;

[0030] Diseases affecting sorghum plants include, but are not limited to, bacterial leaf spot; bacterial leaf streak; bacterial leaf stripe; acremonium wilt; anthracnose; charcoal rot; crazy top downy mildew; damping-off and seed rot; ergot; fusarium head blight, root and stalk rot; grain storage mold; gray leaf spot; latter leaf spot; leaf blight; milo disease; oval leaf spot; pokkah boeng; pythium root rot; rough leaf spot; rust; pythium seedling blight and root rot; smut (covered kernel); smut (head); smut (loose kernel); sooty stripe; downy mildew; tar spot; target leaf spot; and zonate leaf spot and sheath blight.

[0031] Disease resistant plants can have 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% increased resistance compared to control plants. In some embodiments, plants can have 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% increased plant health in the presence of disease compared to control plants.

[0032] As used herein, the term "chromosomal interval" refers to a continuous linear span of genomic DNA present on a single chromosome of a plant. Genetic elements or genes located in a single chromosomal interval are physically linked. The size of a chromosomal interval is not particularly limited. In some aspects, genetic elements located within a single chromosomal interval are genetically linked, typically having a genetic recombination distance of, for example, less than or equal to 20 cM, or alternatively, less than or equal to 10 cM. ​​That is, two genetic elements within a single chromosomal interval recombine at a frequency of less than or equal to 20% or 10%.

[0033] In this application, the phrase "tightly linked" means that recombination between two linked loci occurs at a frequency of about 10% or less (i.e., separated by no more than 10 cM on a genetic map). In other words, the tightly linked loci have at least a 90% chance of co-segregating. When a marker locus shows a significant probability of co-segregating (linking) with a desired trait (e.g., resistance to southern corn rust), the marker locus is particularly useful for the subject matter of the present disclosure. Tightly linked loci (e.g., a marker locus and a second locus) can exhibit an inter-locus recombination frequency of 10% or less, preferably about 9% or less, even more preferably about 8% or less, yet more preferably about 7% or less, yet more preferably about 6% or less, yet more preferably about 5% or less, yet more preferably about 4% or less, yet more preferably about 3% or less, and yet more preferably about 2% or less. In a very preferred embodiment, the related locus shows about 1% or lower, for example about 0.75% or lower, more preferably about 0.5% or lower or more preferably about 0.25% or lower recombination frequency.Be positioned at the same chromosome and have and make the reorganization between the two loci be also considered to each other " adjacent " with less than 10% (for example, about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or still less) frequency distance of the frequency generation.In some cases, two different marks can have identical genetic map coordinate.In this case, the two marks are very adjacent to each other, so that the reorganization between the two occurs with this low frequency of undetectable.

[0034] The term "hybridized" or "crossing" refers to sexual crossing and involves the fusion of two haploid gametes by pollination to produce a diploid offspring (e.g., cells, seeds, or plants). The term encompasses both pollination of one plant by another and selfing (or self-pollination, e.g., when the pollen and ovules are from the same plant).

[0035] An "elite line" is any line produced by breeding for the expression of superior agronomic traits.

[0036] An "exotic variety," "tropical line," or "exotic germplasm" is a variety derived from a plant that is not an available elite line or germplasm variety. In the case of a cross between two plant or germplasm varieties, the offspring of the exotic germplasm are not closely related to the elite germplasm with which it was crossed. Most commonly, the exotic germplasm is not derived from any known elite line but is selected to introduce new genetic elements (usually new alleles) into a breeding program.

[0037] A "favorable allele" is an allele at a specific locus (marker, QTL, gene, etc.) that confers or contributes to an agronomically desirable phenotype (e.g., disease resistance) and allows identification of plants having that agronomically desirable phenotype. A favorable allele of a marker is a marker allele that segregates with the favorable phenotype.

[0038] "Genetic marker" is a nucleic acid that is polymorphic in a population, and the alleles of the genetic marker can be detected and distinguished by one or more analytical methods (such as RFLP, AFLP, isozymes, SNP, SSR, etc.). The term also refers to a nucleic acid sequence that is complementary to the genomic sequence (such as nucleic acid) used as a probe. The mark corresponding to the genetic polymorphism between the colony members can be detected by methods generally recognized in the art. These methods include, for example, sequence-specific amplification methods based on PCR, restriction fragment length polymorphism detection (RFLP), isozyme marker detection, polynucleotide polymorphism detection by allele-specific hybridization (ASH), amplification variable sequence detection of plant genomes, autonomous sequence replication detection, simple repeat sequence detection (SSR), single nucleotide polymorphism detection (SNP), or amplified fragment length polymorphism detection (AFLP). Known generally recognized methods are also used to detect expressed sequence tags (ESTs) and SSR markers derived from EST sequences, as well as randomly amplified polymorphic DNA (RAPD).

[0039] " germplasm " refers to genetic material, and it belongs to or comes from individual (for example, plant), individual group (for example, plant strain, kind or family) or from clone of strain, kind, species or culture, or more generally, whole individual of a certain species or a plurality of species (for example, maize germplasm collection (maize germplasm collection) or Andean germplasm collection (Andean germplasmcollection)).Described germplasm can be the part of organism or cell, or can be separated from described organism or cell.Generally speaking, germplasm provides genetic material with specific molecular composition, and described specific molecular composition provides physical basis for some or all genetic qualities of organism or cell culture.As used herein, germplasm comprises the cell, seed or tissue that can grow out new plant thus, or can be cultivated into the plant part of complete plant, for example leaf, stem, pollen or cell.

[0040] A "haplotype" is an individual's genotype at multiple genetic loci, ie, a combination of alleles. Typically, the genetic loci described by a haplotype are physically and genetically linked, ie, on the same chromosome segment.

[0041] The term "heterogeneity" is used to indicate that individuals within the group differ in their genotype at one or more specific loci.

[0042] The heterotic response or "heterosis" of a material can be defined by its performance above the average of a parent (or high parent) when crossed to an otherwise dissimilar or unrelated group.

[0043] A "heterotic group" includes a group of genotypes that perform well when crossed with genotypes from a different heterotic group (Hallauer et al. (1998) Corn breeding, pp. 463-564, in G.F. Sprague and J.W. Dudley, eds., Corn and corn improvement). Inbred lines are divided into heterotic groups and further subdivided into families within the heterotic groups based on several criteria, such as pedigree, molecular marker-based association, and performance in hybrid combinations (Smith et al. (1990) Theor. Appl. Gen. 80: 833-840). In the United States, the two most widely used heterotic groups are called "Iowa Stiff Stalk Synthetic" (also referred to herein as "rigid stalk") and "Lancaster" or "Lancaster Sure Crop" (sometimes referred to as NSS or non-rigid stalk).

[0044] Some heterotic groups possess the proterties required for becoming female parents, and others possess the proterties required for becoming male parents. For example, in maize, the yield results of the public inbred lines released from a group called BSSS (Iowa rigid stalk synthesis group) have caused these inbred lines and their derivatives to become female pools in the central corn belt. BSSS inbred lines have been hybridized with other inbred lines (e.g., SD 105 and Maiz Amargo), and the general group of this material is known for rigid stalk synthesis (Stiff Stalk Synthetics, SSS), even though not all inbred lines are derived from original BSSS population (Mikel and Dudley, (2006) Crop Sci [Crop Science]: 46: 1193-1205). By default, all other inbred lines that combine well with the SSS inbred lines are assigned to the male pool and are named NSS, i.e., non-rigid stalks, for lack of a better name. This group includes several major heterotic groups such as Lancaster Surecrop, Iodent and Leaming Corn.

[0045] The term "homogeneity" means that members of a group have the same genotype at one or more specific loci.

[0046] The term "hybrid" refers to the offspring obtained from a cross between at least two genetically distinct parents.

[0047] The term "inbred line" refers to a line that has been bred to achieve genetic homogeneity.

[0048] The term "indel" refers to an insertion or deletion, wherein one strain can be referred to as having an inserted nucleotide or DNA fragment relative to a second strain, or the second strain can be referred to as having a deleted nucleotide or DNA fragment relative to the first strain.

[0049] The term "infiltration" refers to the phenomenon that the desired allelomorph of the genetic locus is passed to another kind of genetic background from a kind of genetic background.For example, the desired allelomorph infiltration at the specified locus can be passed to at least one filial generation via the sexual hybridization between two parents of the same species, wherein at least one of the parents has described required allelomorph in its genome. Alternatively, for example allelic transmission can occur by the reorganization between two donor genomes, for example in the fusion protoplast, wherein at least one of them donor protoplast has desired allelomorph in its genome. Required allelomorph can, for example, detect at QTL, transgenic etc. by the mark associated with phenotype. In any case, the offspring that comprises described required allelomorph can be repeatedly backcrossed with the strain with required genetic background and select required allelomorph, to produce the allelomorph that is fixed in the genetic background of selection.

[0050] When the "introgression" is repeated two or more times, the process is often called "backcrossing."

[0051] A "line" or "variety" is a group of individuals having the same parents, which are usually inbred to some extent and are usually homozygous and homogeneous (isogenic or near isogenic) at most gene loci. A "subline" refers to a subpopulation of inbred lines that is genetically different from other similar subpopulations of inbred lines that originated from the same ancestor.

[0052] As used herein, term " linkage " is used to describe the degree of association of a kind of marker locus and another kind of marker locus or some other loci.The linkage relationship between molecular marker and the locus affecting phenotype is represented with " probability " or " probability of adjustment ".Linkage can be expressed as required restriction or scope.For example, in some embodiments, when any mark and any other mark are separated by less than 50,40,30,25,20 or 15 map distance units (or cM) on a single meiotic map (based on the genetic map of the colony (for example, F2) that has carried out a round of meiosis; IBM2 map is made up of multiple meiosis), the mark is linked (hereditarily or physically).In some respects, it is advantageous to limit the linkage range of bracketing, for example, between 10cM and 20cM, between 10cM and 30cM or between 10cM and 40cM.The tighter the linkage of mark and the second locus, the better the indication effect of mark to the second locus. Thus, "closely linked loci," such as a marker locus and a second locus, exhibit a recombination frequency between the loci of 10% or less, preferably about 9% or less, even more preferably about 8% or less, yet more preferably about 7% or less, even more preferably about 6% or less, yet more preferably about 5% or less, even more preferably about 4% or less, yet more preferably about 3% or less, and even more preferably about 2% or less. In a very preferred embodiment, the associated loci exhibit a recombination frequency of about 1% or less, such as about 0.75% or less, more preferably about 0.5% or less, or yet more preferably about 0.25% or less. Two loci that are located on the same chromosome and have a distance such that recombination between the two loci occurs at a frequency of less than 10% (e.g., about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or less) are also considered to be "adjacent" to each other. Because one cM is the distance between two markers that exhibit a recombination frequency of 1%, any marker is tightly linked (genetically and physically) to any other marker that is closely adjacent (e.g., at a distance equal to or less than 10 cM). Two tightly linked markers on the same chromosome can be located 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, or 0.25 cM or closer to each other.

[0053] The term "linkage disequilibrium" refers to the non-random separation of genetic loci or proterties (or both). In either case, linkage disequilibrium means that the relevant loci are physically close enough along a section of chromosome so that they separate together with a frequency higher than random (i.e., non-random). The markers showing linkage disequilibrium are considered to be linked. The loci of linkage have more than 50% chance (such as approximately 51% to approximately 100%) to separate altogether. In other words, two markers of cosegregation have a recombination frequency less than 50% (and, by definition, separated less than 50cM on the same linkage group). As used herein, linkage can be present between two markers, or alternatively, between marker and the locus affecting the phenotype. Marker locus can be " associated " (linkage) with proterties. The linkage degree of marker locus and the locus affecting phenotypic traits is for example measured by the statistical probability (such as, F statistics or LOD scores) of the cosegregation of this molecular marker and the phenotype.

[0054] Linkage disequilibrium is most commonly measured using r 2 Evaluation, the measure r 2 The calculation was performed using the formula in the following literature: Hill, WG and Robertson, A, Theor. Appl. Genet. [Theoretical and Applied Genetics] 38: 226-231 (1968). 2 =1, there is complete LD between the two marker loci, meaning that the markers have not yet undergone recombination segregation and have the same allele frequency. 2 The value depends on the population used. 2 Values ​​greater than 1 / 3 indicate sufficiently strong LD for mapping (Ardlie et al., Nature Reviews Genetics 3:299-309 (2002)). 2 When the value is greater than or equal to 0.33, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1.0, the alleles are in linkage disequilibrium.

[0055] As used herein, "linkage equilibrium" describes a situation in which two markers segregate independently, ie, randomly distribute among progeny. Markers that exhibit linkage equilibrium are considered unlinked (regardless of whether they are located on the same chromosome).

[0056] A "locus" is a position on a chromosome, eg, where a nucleotide, gene, sequence, or marker is located.

[0057] "Logarithm of advantage (LOD) value" or "LOD score" (Risch, Science 255:803-804 (1992)) is used for genetic interval mapping to describe the degree of linkage between two marker loci. A LOD score of three between two markers indicates that the probability of linkage is 1000 times higher than the probability of no linkage, while a LOD score of two indicates that the probability of linkage is 100 times higher than the probability of no linkage. An LOD score greater than or equal to two can be used to detect linkage. The LOD score can also be used to display the strength of association between a marker locus and a quantitative trait in a "quantitative trait locus" mapping. In this case, the LOD score size depends on the tightness between the marker locus and the locus that affects the quantitative trait, as well as the size of the quantitative trait effect.

[0058] The term "plant" includes whole plants, plant cells, plant protoplasts, plant cell or tissue cultures from which plants can be regenerated, plant callus, plant clumps, and intact plant cells or plant parts in plants, such as seeds, flowers, cotyledons, leaves, stems, shoots, roots, root tips, etc. As used herein, "modified plants" means any plant that has a genetic change due to human intervention. The modified plant can have a genetic change introduced by plant transformation, genome editing, or conventional plant breeding.

[0059] " mark " is to find the position on heredity or physical map or to find the mode of linkage between mark and trait locus (locus affecting trait).The position that mark detects can be known by detecting polymorphic allele and genetic location thereof, or is known by hybridizing, sequence matching or amplification of the sequence having carried out physical mapping.Mark can be dna marker (detecting DNA polymorphism), protein (detecting the variation of coded polypeptide) or the phenotype (such as " waxy " phenotype) of simple inheritance.Can be from genomic nucleotide sequence or from expressed nucleotide sequence (for example, from RNA or cDNA of splicing) development dna marker.According to DNA marking technology, described mark is made up of the complementary primer of side joint in described locus and / or the complementary probe of hybridizing with the polymorphic allele of described locus.DNA marker or genetic marker can also be used for describing gene, dna sequence or nucleotide (rather than the component for detecting this gene or dna sequence) on this chromosome itself, and it is usually used (for example breast cancer marker) when this dna marker is associated with the specific proteus in people's genetics. The term marker locus is the locus (gene, sequence or nucleotide) that the marker detects.

[0060] The mark of the genetic polymorphism between the detection group member is generally recognized in the art.The mark can be defined by the type of the polymorphism detected by it and the labeling technology for detecting the polymorphism.Marker types include but are not limited to: restriction fragment length polymorphism detection (RFLP), isozyme marker detection, randomly amplified polymorphic DNA (RAPD), amplified fragment length polymorphism detection (AFLP), simple repeat sequence detection (SSR), the amplification variable sequence detection of plant genome, autonomous sequence replication detection or single nucleotide polymorphism detection (SNP).SNP can be by, for example, by DNA sequencing, PCR-based sequence specific amplification method, the polynucleotide polymorphism detection carried out by allele specific hybridization (ASH), dynamic allele specific hybridization (DASH), molecular beacons, microarray hybridization, oligonucleotide ligase analysis, Flap endonuclease, 5 ' endonuclease, primer extension, single strand conformation polymorphism (SSCP) or temperature gradient gel electrophoresis (TGGE) detect.DNA sequencing (such as pyrophosphate sequencing technology) has the advantage of being able to detect a series of linked SNP alleles that form haplotype. Haplotypes tend to be more informative (detect higher levels of polymorphism) than SNPs.

[0061] A "marker allele," alternatively an "allele of a marker locus," can refer to one of a plurality of polymorphic nucleotide sequences found at a marker locus in a population.

[0062] "Marker-assisted selection" (MAS) is a method of selecting individual plants based on their marker genotype.

[0063] "Marker-assisted counter-selection" is a method whereby marker genotypes are used to identify plants that are not to be selected, allowing them to be removed from a breeding program or planting.

[0064] A "marker haplotype" refers to the combination of alleles at a marker locus.

[0065] "Marker locus" is a specific chromosomal location in a species genome where specific markers can be found. The marker locus can be used to track the presence of a second linked locus (e.g., a linked locus that affects phenotypic trait expression). For example, the marker locus can be used to monitor the separation of alleles at a genetically or physically linked locus.

[0066] As mentioned above, when identifying linked loci, the term "molecular marker" can be used to refer to genetic markers, or its coded product (for example, protein) as a reference point. Mark can be derived from a genomic nucleotide sequence or from an expressed nucleotide sequence (for example, from the RNA, cDNA of splicing, etc.), or from a coded polypeptide. The term also refers to a nucleic acid sequence complementary to a marker sequence or to its side joint, such as a nucleic acid used as a probe or a primer pair capable of amplifying the marker sequence. A "molecular marker probe" is a nucleic acid sequence or molecule that can be used to identify the presence or absence of a marker locus, for example, a nucleic acid probe complementary to a marker locus sequence. Alternatively, in some aspects, a marker probe refers to a probe that can distinguish any type (i.e., genotype) of a specific allele present at a marker locus. When nucleic acid specifically hybridizes in a solution, they are "complementary." When positioned at an insertion / deletion region, for example, a non-collinear region as described herein, some of the markers described herein are also referred to as hybridization markers. This is because, by definition, the insertion region is a polymorphism about a plant without the insertion. Therefore, the marker only needs to indicate whether the insertion / deletion region exists. Any suitable marker detection technology can be used to identify such hybridization markers, such as the use of SNP technology in the examples provided herein.

[0067] An allele is "negatively" associated with a trait when the allele is linked to the trait and when the presence of the allele is an indication that the desired trait or form of the trait will not appear in a plant containing the allele.

[0068] The term "phenotype", "phenotypic trait" or "trait" can refer to the observable expression of a gene or gene series. The phenotype can be observable to the naked eye, or by any other evaluation method known in the art (e.g., weighing, counting, measuring (length, width, angle, etc.), microscopy, biochemical analysis or electromechanical determination). In some cases, the phenotype is directly controlled by a single gene or genetic locus, that is, a "monogenic trait" or "simple genetic trait". In the absence of large-scale environmental changes, monogenic traits can be separated in a population to give a "quality" or "discrete" distribution, that is, the phenotype is attributed to a discrete category. In other cases, the phenotype is the result of multiple genes and can be considered as a "polygenic trait" or "complex trait". Polygenic traits are separated in a population to give a "quantity" or "continuous" distribution, that is, the phenotype cannot be separated into discrete categories. Both monogenic traits and polygenic traits can be affected by the environment in which they are expressed, but polygenic traits tend to have a larger environmental component.

[0069] A "physical map" of a genome is a map showing the linear order of identifiable markers (including genes, markers, etc.) on chromosomal DNA. However, in contrast to a genetic map, the distances between markers are absolute (e.g., measured in base pairs or isolated and overlapping contiguous gene segments) and are not based on genetic recombination (which can vary in different populations).

[0070] A "polymorphism" is a variation in the DNA between two or more individuals within a population. A polymorphism preferably has a frequency of at least 1% in a population. Useful polymorphisms can include single nucleotide polymorphisms (SNPs), simple sequence repeats (SSRs), or insertion / deletion polymorphisms (also referred to herein as "indels").

[0071] A "production marker" or "production SNP marker" is a marker that has been developed for high throughput purposes. Production SNP markers are developed to detect specific polymorphisms and are designed for use with a variety of chemistries and platforms.

[0072] The term "quantitative trait locus" or "QTL" refers to a region of DNA that is associated with the differential expression of a quantitative phenotypic trait in at least one genetic background (e.g., in at least one breeding population). The region of a QTL encompasses or is tightly linked to one or more genes that affect the trait in question.

[0073] A "reference sequence" or "consensus sequence" is a defined sequence used as the basis for sequence alignment. A labeled reference sequence is obtained by sequencing multiple lines at the locus, aligning these nucleotide sequences in a sequence alignment program (e.g., Sequencher), and then obtaining the most common nucleotide sequence of the alignment. Polymorphisms found in these individual sequences are annotated in the consensus sequence. A reference sequence is generally not an exact copy of any individual DNA sequence, but rather represents a mixture of available sequences and is used to design primers and probes for polymorphisms within the sequence.

[0074] An "unfavorable allele" of a marker is a marker allele that segregates with the unfavorable plant phenotype, thus providing the benefit of identifying plants that can be removed from a breeding program or planting.

[0075] The term "yield" refers to the productivity per unit area of ​​a particular plant product with commercial value. Yield is affected by both heredity and environmental factors. "Agronomy," "agronomic traits," and "agronomic trait performance" refer to the proterties (and potential genetic elements) of a given plant variety, which contribute to yield during the vegetative period. Individual agronomic traits include vigor of emergence, nutritional potential, stress tolerance, disease resistance or tolerance, herbicide resistance, branching, flowering, seed formation, seed size, seed density, lodging resistance, threshing performance, etc. Therefore, yield is the ultimate apex of all agronomic traits.

[0076] Provided herein are marker loci that show statistically significant cosegregation with disease resistance traits that confer broad resistance to one or more specific diseases. These loci or additional linked loci and detection of resistance genes can be used in marker-assisted selection as part of a breeding program to produce plants resistant to one or more diseases.

[0077] It has been recognized that in many cases a specific genetic locus associated with a specific phenotype (e.g., disease resistance) can be located in the genome of an organism. Plant breeders can advantageously use molecular markers to identify desired individuals by detecting marker alleles that show a statistically significant probability of cosegregating with the desired phenotype, exhibiting linkage disequilibrium. By identifying molecular markers or clusters of molecular markers that cosegregate with the trait of interest, plant breeders can rapidly select for the desired phenotype by selecting the appropriate molecular marker alleles (a method known as marker-assisted selection or MAS).

[0078] Multiple methods as known in the art can be used for detecting molecular markers or molecular marker clusters that are co-segregated with the purpose proterties (for example disease resistance proterties).The basic concept of these methods is to detect the mark of the alternative genotype (or allelotrope) with significantly different average phenotypes.Therefore, the difference size between the alternative genotype (or allelotrope) between the comparative marker locus or the significance level of the described difference.Infer that the proterties gene is located at the position of one or more marks of the genotypic difference with maximum correlation closest.Two methods for detecting the purpose proterties locus like this are: 1) association analysis (i.e. association location) based on colony and 2) traditional linkage analysis.

[0079] Related Positioning

[0080] The degree and pattern of understanding linkage disequilibrium (LD) in the genome are prerequisites for developing effective, in order to identify and draw the association method of quantitative trait locus (QTL).Linkage disequilibrium (LD) refers to the non-random association of alleles in the individual collection.When observing LD in the allele at the locus of linkage, LD is measured as the LD decay in the specific region across chromosome.The scope of LD reflects the recombination history in this region.The average rate of LD decay in the genome can help predict the quantity and the density of the marker required for whole genome association study, and provides the estimated value of the resolution that can be expected.

[0081] Association or LD mapping aims to identify significant genotype-phenotype associations. It has been developed as a powerful tool for fine mapping in outcrossing species such as humans (Corder et al. (1994) "Protective effect of apolipoprotein-E type-2 allele for late-onset Alzheimer-disease," Nat Genet 7: 180-184; Hastbacka et al. (1992) "Linkage disequilibrium mapping in isolated founder populations: diastrophic dysplasia in Finland," Nat Genet 2: 204-211; Kerem et al. (1989) "Identification of the cystic fibrosis gene: genetic analysis [Identification of cystic fibrosis genes: genetic analysis], Science 245:1073-1080) and maize (Remington et al. (2001) "Structure of linkage disequilibrium and phenotype associations in the maize genome," Proc Natl Acad Sci USA 98:11479-11484; Thornsberry et al. (2001) "Dwarf8 polymorphisms associate with variation in flowering time," Nat Genet 28:286-289; reviewed by Flint-Garcia et al. (2003) "Structure of linkage disequilibrium in plants," Annu Rev Plant Biol. [Annual Review of Plant Biology] 54: 357-374), in which recombination between heterozygotes is frequent and leads to rapid decay of LD.In inbred species, where recombination between homozygous genotypes is not genetically detectable, the degree of LD is greater (i.e., larger blocks of linked markers are inherited together) and this greatly increases the power to detect association maps (Wall and Pritchard, (2003) "Haplotype blocks and linkage disequilibrium in the human genome," Nat Rev Genet 4: 587-597).

[0082] The recombination and mutation history of a population is a function of mating habits and the effective size and age of the population. Larger population sizes provide an enhanced probability of detecting recombination, while older populations are generally associated with higher levels of polymorphism, both of which contribute to a significant increase in the observed LD decay rate. On the other hand, smaller effective population sizes, such as those that have experienced a recent genetic bottleneck, tend to exhibit slower LD decay rates, leading to more extensive haplotype conservation (Flint-Garcia et al., (2003) "Structure of linkage disequilibrium in plants," Annu Rev Plant Biol. 54: 357-374).

[0083] Elite breeding lines provide a valuable starting point for association analyses in which quantitative phenotypic scores (e.g., a disease tolerance rating from one to nine for each line) are used (rather than considering only the tolerance versus resistance allele frequency distribution in the type of allele distribution between groups analyzed). The availability of detailed phenotypic performance data collected over many years through breeding programs and the environment of a large number of elite lines provides a valuable data set for genetic marker association mapping analysis. This paves the way for seamless integration between research and application, and takes advantage of historically accumulated data sets. However, understanding the relationship between polymorphism and recombination is useful for developing appropriate strategies for efficiently extracting maximum information from these resources.

[0084] This type of association analysis neither generates nor requires any map data, but is independent of map position. This analysis compares the phenotypic scores of plants with the genotypes at different loci. Subsequently, using the previously determined map locations of these markers, any suitable map (e.g., a composite map) can optionally be used to help visualize the distribution of identified QTL markers and / or QTL marker clusters.

[0085] Traditional linkage analysis

[0086] Traditional linkage analysis is based on the same principle; however, LD is generated by creating a population from a small number of founders. Founders are selected to maximize the level of polymorphism within a structured population, and the level of co-segregation of polymorphic sites with a given phenotype is assessed. A large number of statistical methods have been used to identify significant marker-trait associations. One such method is the interval mapping method (Lander and Botstein, Genetics [Genetics] 121: 185-199 (1989), in which each of many positions along a genetic map (say, in intervals of 1 cM) is tested for the probability that the gene controlling the trait of interest is located at that position. Genotype / phenotype data are used to calculate the LOD score (logarithm of the probability ratio) for each tested position. When the LOD score is greater than a threshold, there is significant evidence that the gene controlling the trait of interest is located at that location on the genetic map (to be between two specific marker loci).

[0087] Provided herein are marker loci that show statistically significant cosegregation with disease resistance traits as determined by traditional linkage analysis and genome-wide association analysis. Detection of these loci or additional linked loci can be used in marker-assisted breeding programs to produce plants with disease resistance.

[0088] Activities in a marker-assisted breeding program may include, but are not limited to: selecting among new breeding populations based on historical genotype and agronomic trait associations to identify which population has the highest frequency of favorable nucleic acid sequences, selecting for favorable nucleic acid sequences in progeny within a breeding population, selecting among parental lines based on predictions of progeny performance, and advancing lines in germplasm improvement activities based on the presence of favorable nucleic acid sequences.

[0089] Chromosome intervals associated with disease resistance traits are provided. Various methods well known in the art can be used to identify chromosomal intervals. The boundaries of such chromosomal intervals are expanded to encompass markers that will be linked to one or more genes controlling the target trait. In other words, the chromosomal interval is expanded so that any marker within the interval (including the end markers that define the boundaries of the interval) can be used as a marker for disease resistance traits.

[0090] Conversely, if, for example, two markers that are very close together show co-segregation with a desired phenotypic trait, it is sometimes unclear whether each of those markers identifies the same gene or two different genes or genes. In any case, knowledge of how many genes are within a particular physical / genomic interval is not necessary for formulating or practicing which are presented in this disclosure.

[0091] The chromosome 7 interval can comprise any marker identified herein as associated with a disease resistance trait, comprising a sequence that is at least 95% identical to SEQ ID NOs: 4-10 or 13-16. Any marker located within these intervals can be used as a marker for disease resistance and can be used in the context of the methods presented herein to identify and / or select plants with disease resistance, regardless of whether the resistance is newly conferred or enhanced compared to control plants. In certain embodiments, markers located upstream and downstream of the ZmMM1 gene location are very closely linked, both genetically and physically, and therefore can be used to select for the ZmMM1 gene for trait introgression and product development.

[0092] Chromosomal intervals can also be defined by markers that are linked to (and show linkage disequilibrium with) disease resistance genes, and r 2 is a common measure of linkage disequilibrium (LD) in the context of association studies. If the LD between a chromosome 7 marker locus in the target interval and another closely adjacent chromosome 7 marker locus is r 2 If the value is greater than 1 / 3 (Ardlie et al., Nature Reviews Genetics 3:299-309 (2002)), the two loci are in linkage disequilibrium with each other.

[0093] A common measure of linkage is the frequency of co-segregation of traits. This can be expressed as a percentage of co-segregation (recombination frequency) or in centimorgans (cM). cM is a unit of measure for genetic recombination frequency. One cM is equivalent to a 1% chance that a trait at one genetic locus will segregate with a trait at another locus due to hybridization in a single generation (meaning there is a 99% chance that these traits will segregate overall). Since chromosomal distance is roughly proportional to the frequency of hybridization events between traits, there is an approximate physical distance that is associated with recombination frequency.

[0094] The marker locus itself is a trait and can be assessed according to standard linkage analysis by tracking the marker locus during segregation. Thus, one cM equals a 1% chance that a marker locus will segregate with another locus due to hybridization in a single generation.

[0095] The closer the marker is to the gene controlling the desired trait, the more effective and advantageous the marker is as an indicator of the desired trait. Tightly linked loci show an inter-locus hybridization frequency of about 10% or less, preferably about 9% or less, also more preferably about 8% or less, again more preferably about 7% or less, also more preferably about 6% or less, again more preferably about 5% or less, also more preferably about 4% or less, again more preferably about 3% or less, and also more preferably about 2% or less. In a highly preferred embodiment, the related loci (e.g., marker loci and target loci) show a recombination frequency of about 1% or less, such as about 0.75% or less, more preferably about 0.5% or less, or again more preferably about 0.25% or less. Therefore, the loci are separated by a distance of about 10 cM, 9 cM, 8 cM, 7 cM, 6 cM, 5 cM, 4 cM, 3 cM, 2 cM, 1 cM, 0.75 cM, 0.5 cM or 0.25 cM or less. In other words, two loci are considered "adjacent" to each other if they are located on the same chromosome and are at a distance such that recombination between the two loci occurs at a frequency of less than 10% (e.g., about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or less).

[0096] Although specific marker alleles can be co-segregated with disease resistance traits, it is important to note that the marker locus does not necessarily cause the expression of the disease resistance phenotype. For example, it is not a requirement that the marker polynucleotide sequence is part of a gene that produces a disease resistance phenotype (e.g., is part of a gene's open reading frame). The association between a specific marker allele and a disease resistance trait is due to the initial "coupling" phase linkage between the marker allele and the allele in the ancestral strain from which the allele originated. Ultimately, hybridization events between the marker and the genetic locus can change this orientation through repeated recombination. For this reason, the favorable marker allele can change based on the linkage phase present in the parent with disease resistance for creating a segregating population. This does not change the fact that the marker can be used to monitor phenotypic segregation. It merely changes which marker allele is considered favorable in a given segregating population.

[0097] The method proposed herein comprises the existence of one or more marker allelotrope associated with disease resistance in the detection plant, and identifies and / or is selected at those marker loci places and has the plant of favorable allele.Mark has been accredited as and is associated with disease resistance proterties in this article, and therefore can be used for predicting the disease resistance in the plant.Any mark (based on the genetic map of single meiosis) in 50cM, 40cM, 30cM, 20cM, 15cM, 10cM, 9cM, 8cM, 7cM, 6cM, 5cM, 4cM, 3cM, 2cM, 1cM, 0.75cM, 0.5cM or 0.25cM also can be used for predicting the disease resistance of plant.

[0098] Molecular markers can be used in a variety of plant breeding applications (e.g., see Staub et al. (1996) Hortscience 31:729-741; Tanksley (1983) Plant Molecular Biology Reporter. 1:3-8). One major area of ​​interest is the use of marker-assisted selection (MAS) to increase the efficiency of backcrossing and introgression. Molecular markers that exhibit linkage to loci that affect a desired phenotypic trait provide a useful tool for selecting traits in plant populations. This is particularly true when phenotypes are difficult to measure. Because DNA marker assays are less labor-intensive and require less physical space than field phenotyping, larger populations can be assayed, increasing the probability of finding recombinants with the target segment moved from the donor line to the recipient line. The tighter the linkage, the more useful the marker, because recombination is less likely to occur between the marker and the gene responsible for the trait, which could result in false positives. Flanking markers reduce the probability of false-positive selections because double recombination events are required. Ideally, the gene itself has a marker so that recombination between the marker and the gene cannot occur. In some embodiments, the methods disclosed herein generate markers in disease resistance genes, wherein the genes are identified by inferring genomic location from clustering or cluster analysis of conserved domains.

[0099] When a gene is introduced by MAS, not only the gene but also the flanking regions are introduced (Gepts. (2002). Crop Sci. 42: 1780-1790). This is called "linkage drag." In cases where the donor plant is very unrelated to the recipient plant, these flanking regions carry additional genes that can encode agronomically undesirable traits. Even after multiple cycles of backcrossing with elite lines, this "linkage drag" can lead to yield reduction or other negative agronomic characteristics. This is sometimes also called "yield drag." The size of the flanking regions can be reduced by additional backcrossing, although this is not always successful because breeders cannot control the size of the region or the recombination breakpoints (Young et al. (1998) Genetics 120: 579-585). In classical breeding, recombination that helps reduce the size of the donor segment is often selected by chance (Tanksley et al. (1989). Biotechnology 7: 257-264). Even after 20 backcrosses of this type, it is expected that quite large donor chromosome fragments still linked to the gene will be selected. However, if markers are used, it is possible to select those rare individuals that have experienced recombination near the gene of concern. In 150 backcross plants, there is a 95% chance that at least one plant will experience hybridization within the 1cM (based on a single meiotic division map distance) of the gene. Markers make it possible to clearly identify these individuals. Using another backcross of 300 plants, there is a 95% hybridization probability within the 1cM single meiotic division map distance on the other side of the gene, thereby producing a segment near the target gene less than 2cM based on a single meiotic division map distance. This can be achieved in two generations using markers, while without markers, an average of 100 generations (see Tanksley et al., the same) are required. When the exact location of a gene is known, the flanking markers around the gene can be used for selecting recombination in different population sizes. For example, in smaller populations, recombination may be expected to occur further away from the gene, thus requiring more distal flanking markers to detect the recombination.

[0100] The main components of implementing MAS are: (i) defining a population in which marker-trait associations are to be determined, which can be a segregating population, or a random or structured population; (ii) monitoring the segregation or association of polymorphic markers with respect to the trait and using statistical methods to determine linkage or association; (iii) defining a set of desired markers based on the results of the statistical analysis, and (iv) using and / or extrapolating this information to the current set of breeding germplasm to enable marker-based selection decisions. The markers described in this disclosure, as well as other marker types, such as SSRs and FLPs, can be used in marker-assisted selection schemes.

[0101] SSRs can be defined as relatively short runs of tandemly repeated DNA that are 6 bp or less in length (Tautz (1989) Nucleic Acid Research 17:6463-6471; Wang et al. (1994) Theoretical and Applied Genetics 88:1-6). Polymorphisms arise due to variations in the number of repeat units, which can be caused by slippage during DNA replication (Levinson and Gutman (1987) Mol Biol Evol 4:203-221). Variations in repeat length can be detected by designing PCR primers to conserved, non-repeat flanking regions (Weber and May (1989) Am J Hum Genet. 44:388-396). Because SSRs are multiallelic, codominant, regenerable, and amenable to high-throughput automation, they are well suited for mapping and MAS (Rafalski et al. (1996) Generating and using DNA markers inplants. In: Non-mammalian genomic analysis: a practical guide. Academic press, pp. 75-135).

[0102] Various types of SSR markers can be generated, and SSR profiles can be obtained by gel electrophoresis of the amplified products. The scoring of marker genotypes is based on the size of the amplified fragments.

[0103] Various types of FLP markers can also be generated. Most commonly, amplification primers are used to generate fragment length polymorphisms. Such FLP markers are similar to SSR markers in many respects, except that the regions amplified by the primers are generally not highly repetitive regions. Usually due to insertions or deletions, the amplified regions or amplicons still have enough variability between germplasms so that the fragments produced by the amplification primers can be distinguished in polymorphic individuals, and it is known that such insertions and deletions often occur in maize (Bhattramakki et al., (2002). Plant Mol Biol [Plant Molecular Biology] 48, 539-547; Rafalski (2002b), supra).

[0104] SNP markers detect single base pair nucleotide substitutions. Of all molecular marker types, SNPs are the most abundant and therefore have the potential to provide the highest genetic map resolution (Bhattramakki et al., 2002 Plant Molecular Biology 48: 539-547). Since SNPs do not require large amounts of DNA and automation of the assay can be direct, SNPs can be assayed in a so-called "ultra-high throughput" manner at even higher throughput levels than SSRs. SNPs also have the potential to be a relatively low-cost system. These three factors together make the use of SNPs in MAS highly attractive. Several methods are available for SNP genotyping, including but not limited to hybridization, primer extension, oligonucleotide ligation, nuclease cleavage, microsequencing, and coded spheres. These methods have been reviewed in Gut (2001) Hum Mutat 17, 475-492; Shi (2001) Clin Chem 47, 164-172; Kwok (2000) Pharmacogenomics 1, 95-100; and Bhattramakki and Rafalski (2001), Discovery and application of single nucleotide polymorphism markers in plants, In: RJ Henry, ed., Plant Genotyping: The DNA Fingerprinting of Plants, CABI Publishing, Wallingford. A wide range of commercially available technologies utilize these and other methods to detect SNPs, including: Masscode.TM. (Qiagen), (Third Wave Technologies) and Invader (Applied Biosystems), (Applied Biosystems, Inc.) and (Illumina).

[0105] Many SNPs within a sequence or across linked sequences can be used to describe the haplotype of any particular genotype (Ching et al., (2002), BMC Genet. [BMC genetics] 3: 19, Gupta et al., 2001, Rafalski (2002b), Plant Science [plant science] 162: 329-333). Haplotypes can be more informative than single SNPs and can describe any particular genotype in more detail. For example, a single SNP may be the allele "T" for a particular line or variety that is disease-resistant, but the allele "T" may also appear in the breeding population used for the recurrent parent. In this case, a haplotype (e.g., a combination of alleles at linked SNP markers) may be more informative. Once a unique haplotype is assigned to a donor chromosome region, the haplotype can be used in that population or any subpopulation thereof to determine whether an individual has a particular gene. See, for example, WO2003054229. The use of automated high-throughput marker detection platforms known to those of ordinary skill in the art makes this method efficient and effective.

[0106] Many marks proposed herein can be easily used as single nucleotide polymorphism (SNP) markers to select ZmMM1 gene.Utilize PCR, primer is used to increase the DNA segment of the individual (preferably inbred line) representing the diversity of target population.PCR product is directly sequenced in one or two directions.The sequence obtained is compared and identified polymorphism.Described polymorphism is not limited to single nucleotide polymorphism (SNP), and includes insertion and deletion, CAPS, SSR and VNTR (variable number of tandem repeats).Specifically, for fine atlas information as herein described, people can be easy to use the information provided herein to obtain other polymorphic SNP (and other marks) in the region amplified by primers disclosed herein.The mark in described atlas region can be hybridized with BAC or other genomic libraries, or is compared electronically with genomic sequence, to find new sequence in the roughly identical location with described mark.

[0107] In addition to the above-mentioned SSR, FLP and SNP, other types of molecular markers are also widely used, including but not limited to: expressed sequence tags (ESTs), SSR markers derived from EST sequences, randomly amplified polymorphic DNA (RAPD) and other nucleic acid-based markers.

[0108] Isozyme profiles and linked morphological traits can also be used indirectly as markers in some cases. Although they do not directly detect DNA differences, they are often affected by specific genetic differences. However, markers for detecting DNA variation are much more numerous and more polymorphic than isozyme or morphological markers (Tanksley (1983) Plant Molecular Biology Reporter: [Plant Molecular Biology Bulletin] 1: 3-8).

[0109] Sequence alignment or overlapping group can also be used to find the sequence of the upstream or downstream of the specific marker listed herein.Then use these new sequences close to mark as herein described to find and develop functionally equivalent marks.For example, different physics and / or genetic maps are compared to locate the equivalent markers that are not described in the present disclosure but are positioned at similar regions.These maps may be in species, or even span other species that carry out heredity or physically compare.

[0110] In general, MAS uses polymorphic markers that have been identified as having a significant likelihood of co-segregation with traits such as SCR disease resistance traits. Such markers are presumed to be located near one or more genes that confer a plant disease resistance phenotype on the map and are considered to be indicative of the desired trait or marker. Plants are tested for the presence of the desired allele in the marker, and plants containing the desired genotype at one or more loci are expected to transfer the desired genotype along with the desired phenotype to their progeny. Therefore, plants with SCR disease resistance can be selected by detecting one or more marker alleles, and in addition, progeny plants from these plants can also be selected. Thus, a plant containing the desired genotype (i.e., a genotype associated with disease resistance) in a given chromosome region is obtained and then hybridized with another plant. The progeny of this hybrid are then genotyped using one or more markers, and progeny plants with the same genotype in a given chromosome region are then selected as having disease resistance.

[0111] SNPs (i.e., SNP haplotypes) can be used alone or in combination to select favorable resistance gene alleles associated with disease resistance. For example, a SNP haplotype at a QTL on chromosome 7 comprises a sequence having at least 95% identity to SEQ ID NOs: 4-10 or 13-16, and any SNP or indel such as Figure 1 shown, or a combination thereof.

[0112] Those skilled in the art will anticipate that additional polymorphic sites may be present at marker loci in and near the chromosomal markers identified by the methods disclosed herein, wherein one or more polymorphic sites are in linkage disequilibrium (LD) with the alleles at one or more of the polymorphic sites in the haplotype, and therefore can be used in marker-assisted selection programs to introgress target gene alleles or target genomic fragments. If the presence of an allele at one of these sites tends to predict the presence of an allele at other sites on the same chromosome, then two specific alleles at different polymorphic sites are considered to be in LD (Stevens, Mol. Diag. [Molecular Diagnosis] 4: 309-17 (1999)). The marker locus can be located within 5 cM, 2 cM, or 1 cM of the disease resistance trait QTL (on a genetic map based on a single meiosis).

[0113] The skilled artisan will appreciate that allele frequencies (and therefore haplotype frequencies) may vary from one germplasm pool to another. Germplasm pools vary due to differences in maturity, heterosis grouping, geographic distribution, etc. Therefore, SNPs and other polymorphisms in certain germplasm pools may not be informative.

[0114] Plants identified, modified and / or selected by any of the above methods are also of interest.

[0115] The present disclosure encompasses ZmMM1 polypeptides. As used herein, "ZmMM1 polypeptide" and "ZmMM1 protein" are used interchangeably to refer to one or more polypeptides having disease resistance activity and are substantially identical to the ZmMM1 polypeptide of any one of SEQ ID NOs: 1-3. A variety of ZmMM1 polypeptides are contemplated.

[0116] As used herein, "substantially identical" refers to amino acid sequences having at least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity. In some embodiments, sequence identity is over the full length of the polypeptide. When used in conjunction with percent sequence identity, the term "about" means + / - 1.0%.

[0117] As used herein, "recombinant protein" refers to a protein that is no longer in its native environment (e.g., in vitro or in a recombinant bacterial or plant host cell); a protein expressed from a polynucleotide that has been edited from its native version; or a protein expressed from a polynucleotide at a different genomic location relative to the native sequence.

[0118] As used herein, "substantially free of cellular material" refers to polypeptides that include protein preparations having less than about 30%, 20%, 10%, or 5% (by dry weight) of non-target proteins (also referred to herein as "contaminating proteins").

[0119] "Fragments" or "biologically active portions" include polypeptide fragments or polynucleotide fragments that comprise a sequence sufficiently identical to a ZmMM1 gene polypeptide or polynucleotide, respectively, and that exhibit disease resistance when expressed in a plant.

[0120] As used herein, "variant" refers to a protein or polypeptide having an amino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the parent amino acid sequence.

[0121] In some embodiments, the ZmMM1 polypeptide comprises at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125 5%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the ZmMM1 polypeptide, wherein the ZmMM1 polypeptide has disease resistance when expressed in a plant.

[0122] Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of a ZmMM1 polypeptide can be prepared by mutations in the DNA. This can also be accomplished by one of several forms of mutagenesis, such as site-specific double-strand break technology and / or directed evolution. In some aspects, the changes encoded in the amino acid sequence will not substantially affect the function of the protein. Such variants will have the desired activity. However, it will be appreciated that the ability of a ZmMM1 polypeptide to confer disease resistance can be improved by applying these techniques to the compositions of the present disclosure.

[0123] Provided are isolated or recombinant nucleic acid molecules comprising a nucleic acid sequence encoding a ZmMM1 polypeptide or a biologically active portion thereof, as well as nucleic acid molecules sufficient for use as hybridization probes to identify nucleic acid molecules encoding proteins having regions of sequence homology. As used herein, the term "nucleic acid molecule" refers to DNA molecules (e.g., recombinant DNA, cDNA, genomic DNA, plasmid DNA, mitochondrial DNA) and RNA molecules (e.g., mRNA) and analogs of DNA or RNA produced using nucleotide analogs. Nucleic acid molecules can be single-stranded or double-stranded, but are preferably double-stranded DNA.

[0124] As used herein, an "isolated" nucleic acid molecule (or DNA) refers to a nucleic acid sequence (or DNA) that is no longer in its natural environment (e.g., in vitro). As used herein, a "recombinant" nucleic acid molecule (or DNA) refers to a nucleic acid sequence (or DNA) in a recombinant bacterial or plant host cell; it has been edited from its native sequence; or it is located at a different position than the native sequence. In some embodiments, an "isolated" or "recombinant" nucleic acid does not contain sequences that are naturally located on the sides of the nucleic acid in the genomic DNA of the organism from which the nucleic acid was derived (i.e., sequences located at the 5' and 3' ends of the nucleic acid) (preferably protein-encoding sequences). For the purposes of this disclosure, "isolated" or "recombinant" excludes isolated chromosomes when used to refer to nucleic acid molecules. For example, in various embodiments, a recombinant nucleic acid molecule encoding a ZmMM1 polypeptide may comprise less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb of nucleic acid sequence that is naturally located on the sides of the nucleic acid molecule in the genomic DNA of the cell from which the nucleic acid was derived.

[0125] In some embodiments, the isolated nucleic acid molecule encoding the ZmMM1 polypeptide has one or more alterations in the nucleic acid sequence compared to a native or genomic nucleic acid sequence. In some embodiments, alterations in the native or genomic nucleic acid sequence include, but are not limited to, alterations in the nucleic acid sequence due to the degeneracy of the genetic code; alterations in the nucleic acid sequence due to amino acid substitutions, insertions, deletions, and / or additions compared to a native or genomic sequence; removal of one or more introns; deletion of one or more upstream or downstream regulatory regions; and deletion of the 5' and / or 3' untranslated regions relative to the genomic nucleic acid sequence. In some embodiments, the nucleic acid molecule encoding the ZmMM1 polypeptide is a non-genomic sequence.

[0126] A variety of polynucleotides encoding ZmMM1 polypeptides or related proteins are contemplated. When operably linked to appropriate promoter, transcription termination and / or polyadenylation sequences, such polynucleotides can be used to produce ZmMM1 polypeptides in host cells. Such polynucleotides can also be used as probes to isolate homologous or substantially homologous polynucleotides encoding ZmMM1 polypeptides or related proteins.

[0127] In some embodiments, the nucleic acid molecule encoding the ZmMM1 polypeptide is a polynucleotide having a sequence as shown in SEQ ID NO: 4-10, and variants, fragments, and complementary sequences thereof. As used herein, "complementary sequence" refers to a nucleic acid sequence that is sufficiently complementary to a given nucleic acid sequence so that it can hybridize with the given nucleic acid sequence to form a stable duplex. As used herein, "polynucleotide sequence variant" refers to a nucleic acid sequence that encodes the same polypeptide except for the degeneracy of the genetic code.

[0128] In some embodiments, the nucleic acid molecule encoding the ZmMM1 polypeptide is a non-genomic nucleic acid sequence. As used herein, a "non-genomic nucleic acid sequence" or "non-genomic nucleic acid molecule" or "non-genomic polynucleotide" refers to a nucleic acid molecule having one or more changes in the nucleic acid sequence compared to a native or genomic nucleic acid sequence. In some embodiments, changes in the native or genomic nucleic acid molecule include, but are not limited to: changes in the nucleic acid sequence due to the degeneracy of the genetic code; optimization of the nucleic acid sequence for expression in plants; changes in the nucleic acid sequence that introduce at least one amino acid substitution, insertion, deletion, and / or addition compared to the native or genomic sequence; removal of one or more introns associated with the genomic nucleic acid sequence; insertion of one or more heterologous introns; deletion of one or more upstream or downstream regulatory regions associated with the genomic nucleic acid sequence; insertion of one or more heterologous upstream or downstream regulatory regions; deletion of the 5' and / or 3' untranslated regions associated with the genomic nucleic acid sequence; insertion of heterologous 5' and / or 3' untranslated regions; and modification of the polyadenylation site. In some embodiments, the non-genomic nucleic acid molecule is a synthetic nucleic acid sequence.

[0129] In some embodiments, the nucleic acid molecule encoding the ZmMM1 polypeptide disclosed herein is a non-genomic polynucleotide having a nucleotide sequence similar to SEQ ID 94%, 95%, 96%, 97%, 98%, 99% or more identical to the nucleic acid sequence of NOs: 4-10, wherein the ZmMM1 polypeptide has disease resistance activity when expressed in plants.

[0130] In some embodiments, the nucleic acid molecule encodes a ZmMM1 polypeptide variant comprising one or more amino acid substitutions to the amino acid sequence of SEQ ID NOs: 1-3.

[0131] Nucleic acid molecules that are fragments of these nucleic acid sequences encoding ZmMM1 polypeptides are also encompassed in the embodiments. "Fragment" is used herein to refer to a portion of a nucleic acid sequence encoding a ZmMM1 polypeptide. A fragment of a nucleic acid sequence can encode a biologically active portion of a ZmMM1 polypeptide, or it can be a fragment that can be used as a hybridization probe or PCR primer using the methods disclosed below. Nucleic acid molecules that are fragments of a nucleic acid sequence encoding a ZmMM1 polypeptide comprise at least about 150, 180, 210, 240, 270, 300, 330, 360, 400, 450, or 500 consecutive nucleotides, or up to the number of nucleotides present in the full-length nucleic acid sequence encoding a ZmMM1 polypeptide identified by the methods disclosed herein, depending on the intended use. "Contiguous nucleotides," as used herein, refers to nucleotide residues that are immediately adjacent to one another. Fragments of the nucleic acid sequences of the embodiments will encode protein fragments that retain the biological activity of the ZmMM1 polypeptide and, therefore, retain disease resistance. As used herein, "retains disease resistance" refers to a polypeptide having at least about 10%, at least about 30%, at least about 50%, at least about 70%, 80%, 90%, 95% or more of the disease resistance of the full-length ZmMM1 polypeptide set forth in SEQ ID NOs: 1-3.

[0132] " Percentage (%) of sequence identity " is determined to be the percentage of amino acid residues or nucleotides in a candidate sequence (query sequence) that are identical to the corresponding amino acid residues or nucleotides in the reference sequence, after aligning the sequences and introducing gaps (if necessary) to achieve maximum percentage sequence identity, and without considering any conservative substitutions of amino acids as part of sequence identity. The comparison for determining the percentage of sequence identity can be achieved in various ways within the technical scope of the art, for example, using publicly available computer software, such as BLAST, BLAST-2. Those skilled in the art can determine the appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment over the full length of the sequences being compared. The percentage of identity between two sequences is a function of the number of identical positions shared by the sequences (e.g., the number of identical positions between the query sequence and the subject sequence / the total number of positions of the query sequence × 100).

[0133] In some embodiments, the ZmMM1 polynucleotide encodes a ZmMM1 polypeptide comprising an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NOs: 1-3 throughout the entire length.

[0134] The embodiments also encompass nucleic acid molecules encoding variants of the ZmMM1 polypeptide. "Variants" of nucleic acid sequences encoding ZmMM1 polypeptides include those sequences encoding ZmMM1 polypeptides identified by the methods disclosed herein but having conservative differences due to the degeneracy of the genetic code, as well as those sequences that are sufficiently identical as described above. Naturally occurring allelic variants can be identified using well-known molecular biology techniques, such as polymerase chain reaction (PCR) and hybridization techniques as outlined below. Variant nucleic acid sequences also include synthetically derived nucleic acid sequences that have been generated, for example, using site-directed mutagenesis, but still encode the ZmMM1 gene polypeptides disclosed herein.

[0135] The skilled artisan will further appreciate that changes can be introduced by mutation of the nucleic acid sequence, thereby resulting in changes in the amino acid sequence of the encoded ZmMM1 polypeptide without altering the biological activity of the protein. Thus, variant nucleic acid molecules can be generated by introducing one or more nucleotide substitutions, additions, and / or deletions into the corresponding nucleic acid sequence disclosed herein, such that one or more amino acid substitutions, additions, or deletions are introduced into the encoded protein. Mutations can be introduced by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Such variant nucleic acid sequences are also encompassed by the present disclosure.

[0136] Alternatively, variant nucleic acid sequences can be prepared by randomly introducing mutations along all or part of the coding sequence (e.g., by saturation mutagenesis), and the resulting mutants can be screened for their ability to confer activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be recombinantly expressed, and the activity of the protein can be determined using standard assay techniques.

[0137] The polynucleotides of the present disclosure and fragments thereof are optionally used as substrates for various recombination and recursive recombination reactions, in addition to the standard cloning methods described by, for example, Ausubel, Berger, and Sambrook, i.e., to produce other polypeptide homologs and fragments thereof with desired properties. Various such reactions are known. Methods for producing variants of any nucleic acid listed herein (these methods include recursively recombining such polynucleotides with a second (or more) polynucleotides to form a variant polynucleotide library) are also embodiments of the present disclosure, as are the libraries produced, the cells comprising such libraries, and any recombinant polynucleotides produced by such methods. Additionally, such methods optionally include selecting variant polynucleotides from such libraries based on activity, as where such recursive recombination is performed in vitro or in vivo.

[0138] Various diversity generation schemes (including nucleic acid recursive recombination schemes) are available and are fully described in the art. The programs can be used alone and / or in combination to generate one or more variants of a nucleic acid or nucleic acid set, as well as variants of an encoded protein. Individually or collectively, these programs provide robust and widely applicable methods for generating diverse nucleic acids and nucleic acid sets (including, for example, nucleic acid libraries) that can be used, for example, to engineer or rapidly evolve nucleic acids, proteins, pathways, cells, and / or organisms with new and / or improved characteristics.

[0139] Although for the sake of clarity, distinctions and classifications are made in the subsequent discussion, it should be understood that the techniques are generally not mutually exclusive. In fact, various methods can be used alone or in combination, in parallel or in series, to obtain different sequence variants.

[0140] The result of any diversity generation program described herein can be the generation of one or more nucleic acids, which can select or screen for nucleic acids having or conferring desired characteristics or nucleic acids encoding proteins having or conferring desired characteristics. After diversification by one or more methods otherwise available herein or by the skilled person, any nucleic acid produced can be selected for desired activity or characteristic, such as this activity at the desired pH. This can include identifying any activity that can be detected, for example, in an automated or automated form, by any assay in the art. Various related (or even unrelated) characteristics can be evaluated in series or in parallel, as appropriate, by the practitioner.

[0141] The nucleotide sequence of embodiment can also be used for separating corresponding sequence from different sources.In this way, such sequence (based on the sequence homology of itself and the sequence identified by the method disclosed herein) can be identified using methods such as PCR, hybridization.The embodiment contains the sequence selected based on the sequence identity with the whole sequence shown in this article or its fragment.This sequence includes the sequence of the straight homologue as described sequence.Term " straight homologue " refers to the gene that is derived from common ancestor gene and is found in different species due to species formation.When its nucleotide sequence and / or its encoded protein sequence have the basic identity as defined elsewhere herein, the gene found in different species is considered to be straight homologue.

[0142] In the PCR method, oligonucleotide primers can be designed for use in a PCR reaction to amplify the corresponding DNA sequence from cDNA or genomic DNA extracted from any organism of interest. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed in Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Plainview, New York), hereinafter "Sambrook". See also, Innis et al., eds., (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press, New York); Innis and Gelfand, eds., (1995) PCR Strategies (Academic Press, New York); and Innis and Gelfand, eds., (1999) PCR Methods Manual (Academic Press, New York). Known PCR methods include, but are not limited to, methods using paired primers, nested primers, single-specific primers, degenerate primers, gene-specific primers, vector-specific primers, partially mismatched primers, and the like.

[0143] In hybridization methods, all or part of the nucleic acid sequence can be used to screen cDNA or genomic libraries. Methods for constructing such cDNA and genomic libraries are generally known in the art and are disclosed in Sambrook and Russell, (2001), supra. So-called hybridization probes can be genomic DNA fragments, cDNA fragments, RNA fragments or other oligonucleotides and can be labeled with a detectable group (such as 32P or any other detectable label, such as other radioisotopes, fluorescent compounds, enzymes or enzyme cofactors). Probes for hybridization can be prepared by labeling synthetic oligonucleotides based on the nucleic acid sequence encoding a known polypeptide disclosed herein. Degenerate primers can be used in addition, which are designed based on conserved nucleotides or amino acid residues in the nucleic acid sequence or the encoded amino acid sequence. Such probes typically comprise a region of the following nucleic acid sequence that hybridizes under stringent conditions with at least about 12, at least about 25, at least about 50, 75, 100, 125, 150, 175 or 200 consecutive nucleic acids of the nucleic acid sequence encoding the polypeptide or its fragment or variant. Methods for preparing probes for hybridization and stringency conditions are generally known in the art and are disclosed in Sambrook and Russell, (2001), supra.

[0144] The use of the term "nucleotide construct" herein is not intended to limit the embodiments to nucleotide constructs comprising DNA. Those of ordinary skill in the art will recognize that nucleotide constructs, particularly polynucleotides and oligonucleotides consisting of ribonucleotides and combinations of ribonucleotides and deoxyribonucleotides, can also be used in the methods disclosed herein. The nucleotide constructs, nucleic acids, and nucleotide sequences of the embodiments additionally encompass all complementary forms of such constructs, molecules, and sequences. In addition, the nucleotide constructs, nucleic acid molecules, and nucleotide sequences of the embodiments encompass all nucleotide constructs, molecules, and sequences that can be used for the plant transformation methods of the embodiments, including but not limited to those consisting of deoxyribonucleotides, ribonucleotides, and combinations thereof. Such deoxyribonucleotides and ribonucleotides include both naturally occurring molecules and synthetic analogs. The nucleotide constructs, nucleic acids, and nucleotide sequences of the embodiments also encompass all forms of nucleotide constructs, including but not limited to single-stranded forms, double-stranded forms, hairpins, stem-loop structures, etc.

[0145] Additional embodiments relate to transformed organisms, such as organisms selected from the group consisting of plant cells, bacteria, yeast, baculovirus, protozoa, nematodes, and algae, wherein the transformed organism comprises a DNA molecule of the embodiments, an expression cassette comprising the DNA molecule, or a vector comprising the expression cassette, which is stably incorporated into the genome of the transformed organism.

[0146] The sequence of the embodiment is provided in a DNA construct for expression in the target organism. The construct will include 5' and 3' regulatory sequences operably connected to the sequence of the embodiment. As used herein, the term "operably connected" refers to a functional connection between a promoter and a second sequence, wherein the promoter sequence initiates and mediates transcription of the DNA sequence corresponding to the second sequence. Generally, being operably connected means that the connected nucleic acid sequence is continuous and, if necessary, connects two protein coding regions in the same reading frame. The construct can additionally contain at least one other gene to be co-transformed into an organism. Alternatively, one or more other genes can be provided on multiple DNA constructs.

[0147] The DNA construct is provided with a plurality of restriction sites for inserting the polypeptide gene sequence of the present disclosure, which will be under the transcriptional regulation of the regulatory region. The DNA construct may additionally contain a selectable marker gene.

[0148] According to the transcription direction of 5' to 3', the DNA construct will generally include: a transcription and translation initiation region (i.e., a promoter), the DNA sequence of the embodiment, and a transcription and translation termination region (i.e., a termination region) that is functional in the organism used as the host. For the host organism and / or sequence of the embodiment, the transcription initiation region (i.e., a promoter) can be natural, similar, exogenous, or heterologous. In addition, the promoter can be a native sequence, or alternatively, a synthetic sequence. As used herein, the term "exogenous" means that the promoter is not found in the natural organism into which the promoter is introduced. When the promoter is "exogenous" or "heterologous" for the sequence of the embodiment, it refers to a promoter that is not a natural or naturally occurring promoter for the operably connected sequence of the embodiment. As used herein, a chimeric gene comprises a coding sequence operably connected to a transcription initiation region, and the transcription initiation region is heterologous to the coding sequence. When the promoter is a native (native or natural) sequence, the expression of the operably connected sequence changes from wild-type expression, which results in a change in phenotype.

[0149] In some embodiments, the DNA construct comprises a polynucleotide encoding a ZmMM1 polypeptide of the embodiments. In some embodiments, the DNA construct comprises a polynucleotide encoding a fusion protein comprising a ZmMM1 polypeptide of the embodiments.

[0150] In some embodiments, the DNA construct may further include a transcription enhancer sequence. As used herein, the term "enhancer" refers to a DNA sequence that can stimulate promoter activity and may be an innate element or heterologous element inserted to enhance the level or tissue specificity of a promoter. Various enhancers are known in the art, including, for example, introns with gene expression enhancing properties in plants (U.S. Patent Application Publication No. 2009 / 0144863), the ubiquitin intron (i.e., maize ubiquitin intron 1 (see, e.g., NCBI sequence S94464)), the ω enhancer or ω major enhancer (Gallie et al., (1989) Molecular Biology of RNA, ed. Cech (Liss, New York) 237-256 and Gallie et al., (1987) Gene 60:217-25), the CaMV 35S enhancer (see, e.g., Benfey et al., (1990) EMBO J. 9:1685-96), and the enhancers of U.S. Patent No. 7,803,992 can also be used. The above list of transcriptional enhancers is not meant to be limiting. Any suitable transcriptional enhancer can be used in the embodiments.

[0151] The termination region may be native to the transcriptional initiation region, may be native to the operably linked DNA sequence of interest, may be native to the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, sequence of interest, plant host, or any combination thereof).

[0152] Convenient termination regions are available from the Ti-plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al., (1991) Mol. Geh. Genet. 262:141-144; Proudfoot, (1991) Cell 64:671-674; Sanfacon et al., (1991) Genes Dev. 5:141-149; Mogen et al., (1990) Plant Cell 2:1261-1272; Munroe et al., (1990) Gene 91:151-158; Ballas et al., (1989) Nucleic Acids Res. 17:7891-7903 and Joshi et al., (1987) Nucleic Acids Res. 15:9627-9639.

[0153] Where appropriate, nucleic acids can be optimized to increase expression in the host organism. Thus, where the host organism is a plant, synthetic nucleic acids can be synthesized using plant-preferred codons to improve expression. For a discussion of host preference usage, see, for example, Campbell and Gowri, (1990) Plant Physiol. 92: 1-11. For example, although the nucleic acid sequences of the embodiments are expressed in both monocotyledonous and dicotyledonous plant species, the sequences can be modified to take into account the specific preferences and GC content preferences of monocotyledonous or dicotyledonous plants, as these preferences have been shown to differ (Murray et al. (1989) Nucleic Acids Res. 17: 477-498). Thus, plant preferences for specific amino acids can be derived from known gene sequences of plants.

[0154] It is known that there are other sequence modifications that can enhance gene expression in cellular hosts. These include eliminating sequences encoding false polyadenylation signals, sequences encoding exon-intron splice site signals, sequences encoding transposon-like repeats, and other sequences that are well characterized and may be detrimental to gene expression. The GC content of the sequence can be adjusted to the average level of a given cellular host, as calculated by reference to known genes expressed in the host cell. As used herein, the term "host cell" refers to a cell that contains a vector and supports replication and / or expression of an expression vector. The host cell can be a prokaryotic cell such as Escherichia coli, or a eukaryotic cell such as a yeast, insect, amphibian or mammalian cell, or a monocotyledonous or dicotyledonous plant cell. An example of a monocotyledonous host cell is a maize host cell. When possible, the sequence is modified to avoid the occurrence of predictable hairpin secondary mRNA structures.

[0155] In preparing the expression cassette, the various DNA fragments may be manipulated to provide the DNA sequence in the proper orientation and, where appropriate, in the proper reading frame. To this end, adapters or linkers may be employed to connect the DNA fragments, or other manipulations may be involved to provide convenient restriction sites, remove excess DNA, remove restriction sites, etc. For this purpose, in vitro mutagenesis, primer repair, restriction enzyme digestion, annealing, and resubstitution (e.g., conversion and transversion) may be involved.

[0156] Many promoters can be used to implement the embodiments. A promoter can be selected based on the desired results. The nucleic acid can be combined with a constitutive, tissue-preferred, inducible, or other promoter for expression in a host organism.

[0157] The methods of the embodiments involve introducing a polypeptide or polynucleotide into a plant. As used herein, "introducing" means presenting the polynucleotide or polypeptide to the plant in such a manner that the sequence enters the interior of the plant cell. The methods of the embodiments do not depend on the specific method used to introduce one or more polynucleotides or one or more polypeptides into a plant, as long as the polynucleotide or polypeptide enters the interior of at least one cell of the plant. Methods for introducing one or more polynucleotides or one or more polypeptides into a plant are known in the art and include, but are not limited to, stable transformation methods, transient transformation methods, and viral-mediated methods.

[0158] As used herein, "stable transformation" means that the nucleotide construct introduced into a plant is integrated into the genome of the plant and can be inherited by its progeny. As used herein, "transient transformation" means that a polynucleotide is introduced into the plant and is not integrated into the genome of the plant, or that a polypeptide is introduced into the plant. As used herein, "plant" refers to whole plants, plant organs (e.g., leaves, stems, roots, etc.), seeds, plant cells, propagules, and embryos and progeny thereof. Plant cells can be differentiated or undifferentiated (e.g., callus, suspension culture cells, protoplasts, leaf cells, root cells, phloem cells, and pollen).

[0159] Transformation protocols and protocols for introducing nucleotide sequences into plants can vary depending on the type of plant or plant cell (ie, monocot or dicot) being targeted for transformation. Suitable methods for introducing nucleotide sequences into plant cells and subsequent insertion into the plant genome include microinjection (Crossway et al., (1986) Biotechniques 4:320-334), electroporation (Riggs et al., (1986) Proc. Natl. Acad. Sci. USA 83:5602-5606), Agrobacterium-mediated transformation (U.S. Pat. Nos. 5,563,055 and 5,981,840), direct gene transfer (Paszkowski et al., (1984) EMBO J 3:2717-2722), and ballistic particle acceleration (see, e.g., U.S. Pat. Nos. 4,945,050; 5,879,918; 5,886,244 and 5,932,782; Tomes et al., (1995) Plant Cell, Tissue, and Organ Culture: Fundamental Biology). Methods [Plant Cell, Tissue and Organ Culture: Basic Methods], Gamborg and Phillips, eds. (Springer-Verlag, Berlin; and McCabe et al., (1988) Biotechnology 6:923-926); and Lecl transformation method (WO 00 / 28058). For potato transformation methods, see Tu et al., (1998) Plant Molecular Biology 37:829-838 and Chong et al., (2000) Transgenic Research 9:71-78. Additional transformation methods can be found in Weissinger et al. (1988) Ann. Rev. Genet. 22:421-477; Sanford et al. (1987) Particulate Science and Technology 5:27-37 (onion); Christou et al. (1988) Plant Physiol. 87:671-674 (soybean); McCabe et al. (1988) Bio / Technology 6:923-926 (soybean); Finer and McMullen (1991) In Vitro Cell Dev. Biol.[In Vitro Cell Biology and Developmental Biology] 27P:175-182 (soybean); Singh et al., (1998) Theor. Appl. Genet. [Theoretical and Applied Genetics] 96:319-324 (soybean); Datta et al., (1990) Biotechnology [Biotechnology] 8:736-740 (rice); 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 (maize); Klein et al., (1988) Biotechnology [Biotechnology] 6:559-563 (maize); U.S. Patent Nos. 5,240,855; 5,322,783 and 5,324,646; Klein et al., (1988) Plant Physiol. 91:440-444 (maize); Fromm et al. (1990) Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren et al. (1984) Nature (London) 311:763-764; U.S. Pat. No. 5,736,369 (cereals); Bytebier et al. (1987) Proc. Natl. Acad. Sci. USA 84:5345-5349 (Liliaceae); De Wet et al. (1985) The Experimental Manipulation of Ovule Tissues [Experimental Manipulation of Ovule Tissue], Chapman et al., eds. (Longman, New York), pp. 197-209 (pollen); Kaeppler et al., (1990) Plant Cell Reports 9:415-418 and Kaeppler et al., (1992) Theor. Appl. Genet. 84:560-566 (whisker-mediated transformation); D'Halluin et al., (1992) Plant Cell 4:1495-1505 (electroporation); Li et al., (1993) Plant Cell Reports 12:250-255 and Christou and Ford, (1995) Annals of Botany 75:407-413 (rice); Osjoda et al., (1996) Nature Biotechnology [Nature Biotechnology] 14: 745-750 (maize via Agrobacterium tumefaciens).

[0160] In certain embodiments, genome editing techniques can be used to introduce polynucleotide compositions into the genome of a plant, or genome editing techniques can be used to edit polynucleotides previously introduced into the plant genome. For example, the identified polynucleotides can be introduced into the desired position in the plant genome using double-strand break technology (such as TALEN, meganucleases, zinc finger nucleases, CRISPR-Cas, etc.). For example, for the purpose of site-specific insertion, the identified polynucleotides can be introduced into the desired position in the genome using the CRISPR-Cas system. The desired position in the plant genome can be any target site required for insertion, such as a genomic region suitable for breeding, or can be a target site located in a genomic window with an existing purpose trait. Existing purpose traits may be endogenous traits or previously introduced traits.

[0161] In certain embodiments, when disease resistance ZmMM1 gene alleles have been identified in the genome, genome editing techniques can be used to change or modify the polynucleotide sequence. Site-specific modifications in the desired ZmMM1 gene allele polynucleotides can be introduced, including modifications produced using any method for introducing site-specific modifications, including but not limited to using gene repair oligonucleotides (e.g., U.S. Publication 2013 / 0019349), or by using double-strand break technology, such as TALEN, large-range nucleases, zinc finger nucleases, CRISPR-Cas, etc. Such technology can be used to modify the polynucleotides previously introduced by inserting, deleting or replacing nucleotides in the introduced polynucleotides. Alternatively, double-strand break technology can be used to add additional nucleotide sequences to the introduced polynucleotides. Additional sequences that can be added include additional expression elements (e.g., enhancer sequences and promoter sequences). In another embodiment, genome editing techniques can be used to locate additional disease resistance proteins near the ZmMM1 polynucleotide compositions in the genome of the plant to produce molecular stacks of disease resistance proteins.

[0162] "Altered target site," "altered target sequence," "modified target site," and "modified target sequence" are used interchangeably herein and refer to a target sequence as disclosed herein that comprises at least one alteration when compared to an unaltered target sequence. Such "alterations" include, for example: (i) substitution of at least one nucleotide, (ii) deletion of at least one nucleotide, (iii) insertion of at least one nucleotide, or (iv) any combination of (i)-(iii).

[0163] Example

[0164] The following examples are provided to illustrate but not limit the claimed subject matter. 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 present disclosure or the scope of the appended claims.

[0165] Example 1. ZmMM1 regulates the lesion-mimic phenotype in maize

[0166] C117 is a near-isogenic line (NIL) from the BC2F7 population, derived from a single F1 cross between highland teosinte (Zea mays ssp. mexicana) and the maize inbred line Mo17, with Mo17 as the recurrent parent exhibiting a lesion-like phenotype. By mapping the F2 populations from C117 and Mo17, a major QTL controlling the lesion-like phenotype was identified on chromosome 7 and designated qLMchr7. Using a position-based cloning approach, qLMchr7 was fine-mapped to a 5 kb interval flanked by markers M2 (SEQ ID NOs: 27 and 28) and M3 (SEQ ID NOs: 11 and 12) ( Figure 1 There is only one gene annotated within this interval based on the teosinte genome sequence, which is named ZmMM1 (Zea mays Mexicana lesion-like 1). Further fine mapping restricted qLMchr7 to a 1 kb interval 950 bp downstream of the ZmMM1 CDS ( Figure 1 The 1 kb region is part of the ZmMM1 3′UTR, as determined by 3′ RACE analysis. Within the 1 kb qLMchr7 region, there are 20 SNPs and 7 indels between C117 and Mo17 ( Figure 1 The qLMchr7 sequence of C117 was compared with the qLMchr7 sequences of 46 different maize inbred lines, and a specific 30 bp region in C117 (SEQ ID NO: 16; and corresponding to 24 bp in Mo17, SEQ ID NO: 15) was found to have two SNPs and one indel, of which C117 had a unique haplotype ( Figure 1 ).

[0167] Because qLMchr7 is part of the 3′UTR of ZmMM1, the function of qLMchr7 was tested to see whether it is dependent on ZmMM1 and whether qLMchr7 is a regulatory element for ZmMM1 expression. Although the transcript levels of ZmMM1 were similar in Mo17 and C117 leaves, the ZmMM1 protein levels determined by Western blotting with anti-ZmMM1 antibodies were significantly higher in leaves containing the C117 qLMchr7 allele (qLMchr7c117 ) in NILs was higher than that in Mo17 qLMchr7 allele (qLMchr7 Mo17 ). This confirms that qLMchr7 c117 than qLMchr7 Mo17 Produces higher ZmMM1 protein levels.

[0168] In Nicotana benthamiana, transient overexpression of C117 or Mo17 ZmMM1CDS with a 35S promoter resulted in cell death. When a 1 kb qLMchr7 fragment (SEQ ID NOs: 13 and 14) was inserted between the ZmMM1CDS and the terminator sequence, the expression of qLMchr7 with C1 17 still caused cell death in N. benthamiana, whereas only the ZmMM1 construct with qLMchr7 Mo17 Although both alleles of qLMchr7 significantly reduced the transcript and protein levels of ZmMM1 compared with the construct without the qLMchr7 fragment, the construct with qLMchr7 c117 and qLMchr7 Mo17 There was no difference in ZmMM1 transcript levels between the constructs. Mo17 Compared with qLMchr7c 117 The construct produced higher ZmMM1 protein levels, which was consistent with the expression results in maize. In addition, qLMchr7 was replaced with the corresponding 30 bp in C117. Mo17 A 24 bp specific region in the qLMchr7 Mo17-m ) increases ZmMM1 protein levels and causes a strong cell death phenotype in N. benthamiana. In contrast, qLMchr7 was replaced with the corresponding 24 bp in Mo17. C117 A 30 bp specific region in qLMchr7 C117-m ) reduced ZmMM1 protein levels and led to a weak cell death phenotype in N. benthamiana. Therefore, it can be concluded that qLMchr7 regulates ZmMM1 expression at the protein level, and higher ZmMM1 protein levels are associated with the disease-like phenotype in C117.

[0169] A loss-of-function ZmMM1 mutant allele was identified from a B73 EMS-mutagenized population. This mutant allele (zmmm1-1) harbors an unintentional mutation in the second exon of ZmMM1 that introduces a premature stop codon. Overexpression of zmmm1-1 does not cause cell death in Nicotiana benthamiana cells. The zmmm1-1 mutant was crossed with a qLMchr7-bearing strain. c117 Plants with the same allele were crossed and a pair of NILs were identified in the subsequent F3 population. Both NILs had the same qLMchr7 C117 allele (1 kb fragment). However, a wild-type ZmMM1 from C117 (ZmMM1-qLMchr7 C117 ), while the other one had a mutant zmmm1-1 allele (zmmm1-1-qLMchr7 C117 ). ZmMM1-qLMchr7 C117 Plants showed a clear disease-like phenotype, but zmmm1-1-qLMchr7 C117 These observations confirmed that ZmMM1 was responsible for the disease-like phenotype in C117.

[0170] Example 2. ZmMM1 positively regulates the effects of northern leaf blight (NLB, also known as corn leaf spot) and gray leaf spot (GLS, also known as corn leaf spot) on the Gray leaf spot) and southern corn rust (SCR)

[0171] Plants homozygous for the zmmm1-1 mutant were crossed with B73 plants to generate F1 plants and F2 populations. F2 plants harboring the homozygous zmmm1-1 allele were significantly more susceptible to NLB and GLS than F2 plants harboring wild-type ZmMM1, as determined by lesion length on infected plants in the field. Zmmm1-1 mutant F2 plants were also more susceptible to SCR than wild-type F2 plants after inoculation with Puccinia maydis conidia in the greenhouse, as determined by visual comparison of fungal biomass accumulation (qRT-PCR amplification of Puccinia maydis actin mRNA) and endospore abundance. Therefore, it was concluded that knocking out ZmMM1 increases the susceptibility of maize to NLB, GLS, and SCR.

[0172] Two pairs of qLMchr7 carriers were evaluated in the field C117 or qLMchr7 Mo17 Alleles of NIL are resistant to NLB, GLS, and SCR. Disease phenotypes are ranked from 1 to 9, with "1" being the most resistant and "9" being the most susceptible. C117 NIL ratio of alleles with qLMchr7 Mo17The NIL allele was more resistant to NLB, GLS, and SCR. The results showed that plants with the teosinte ZmMM1 allele were more resistant to multiple pathogens than plants with the maize Mo17 ZmMM1 allele.

[0173] Example 3. Identification of ZmMM1 protein target genes

[0174] ZmMM1 (CDS sequence - SEQ ID NO: 9; genomic sequence - SEQ ID NO: 5) encodes a transcription factor containing a MYB DNA-binding domain (SEQ ID NO: 3). Transcriptional activity assays in protoplasts showed that ZmMM1, when fused to the DNA-binding domain (BD) of GAL4, significantly repressed expression of the reporter gene GUS, which contains four GAL4 DNA-binding sites in its promoter, indicating that ZmMM1 is a transcriptional repressor. DNA affinity purification sequencing (DAP-seq) was performed and four candidate ZmMM1 target genes (ZmMT1, ZmMT2, ZmMT3, and ZmMT4) were identified. ChIP-qPCR assays confirmed that the ZmMM1 protein directly bound to the promoter regions of the four target genes. Finally, transient expression of ZmMT3 (SEQ ID NO: 18) in Nicotiana benthamiana suppressed ZmMM1-induced cell death. Because ZmMT1 positively regulates disease resistance and negatively regulates the expression of its target genes, downregulation of ZmMT1 (SEQ ID NOs: 20 and 21), ZmMT2 (SEQ ID NO: 23), ZmMT3, or ZmMT4 (SEQ ID NO: 25) enhances resistance to multiple pathogens. ZmMT1 (SEQ ID NOs: 20 and 21), ZmMT2 (SEQ ID NO: 23), and ZmMT3 (SEQ ID NO: 18) are all long noncoding RNAs (lncRNAs), while ZmMT4 (SEQ ID NO: 25) encodes a polypeptide. Sequence Listing <110> Huazhong Agricultural University Lai Zhibing <120> Methods for identifying, selecting and producing disease-resistant crops <130> RTS22658A <160> 28 <170> PatentIn version 3.5 <210> 1 <211> 369 <212> PRT <213> corn <400> 1 Met Gly Leu Asp Val Met Glu Ile Gly Met Gly Ala Asp Leu Ser Leu 1 5 10 15 Asp Leu Arg His Phe Ala Ser Lys Ala Val Arg Gln Ser Lys Asp Asp 20 25 30 Thr Pro Ala Pro Asp Met Asp Ala Cys Ile Arg Arg Leu Glu Glu Glu 35 40 45 Arg Gly Lys Ile Glu Met Phe Lys Arg Asp Leu Pro Leu Cys Ala Arg 50 55 60 Leu Leu Ala Asp Val Ile Asp Val Met Lys Glu Glu Ala Gly Lys Lys 65 70 75 80 Lys Thr Thr Thr Arg Arg Arg Ser Asp Arg Arg Leu Ala Ser Ala Ala 85 90 95 Ala Asp Asp Glu Glu Glu Glu Ala Asp Gly Ala Thr Ala Asp Lys Ser 100 105 110 Lys Trp Met Ser Thr Ala Gln Leu Trp Thr Gly Asp Ser Gly Arg Glu 115 120 125 Asp Ala Glu Ser Glu Lys Gln Asp Lys Gly Arg Cys Ser Pro Glu Ala 130 135 140 Arg Ser Arg Gly Ala Leu Leu Pro Phe Lys Ala Asp Val Gly Ser Gly 145 150 155 160 Ala Pro Ala Phe Ala Pro Leu Phe Leu Arg Thr Asp Asp Lys Ala Ala 165 170 175 Ala Ala Arg Val Gly Val Pro Asp Leu Ser Ser Leu Leu Ser Pro Pro 180 185 190 Ala Thr Met Pro Pro Ala Asp Ala Gly Ala Glu Glu Ser Arg Arg Gln 195 200 205 Val Val Gly Phe Ala Gln Ala Ala Ala Arg Ala Ala Ala Met Ala Pro 210 215 220 Ser Ala Pro Ala Leu Gly Leu Gln Ser Gln Gln Gln Gln Gln Gln 225 230 235 240 Gln Gln Ala Arg Lys Ala Arg Arg Cys Trp Ser Thr Glu Leu His Arg 245 250 255 Lys Phe Val Ala Ala Leu Asp Gln Leu Gly Gly Pro Gln Val Ala Thr 260 265 270 Pro Lys Gln Ile Arg Glu Leu Met Lys Val Asp Gly Leu Thr Asn Asp 275 280 285 Glu Val Lys Ser His Leu Gln Lys Tyr Arg Leu His Asn Arg Arg Ala 290 295 300 Pro Gly Ser Gly Val Val Arg Gln Pro Ile Val Leu Val Gly Gly Leu 305 310 315 320 Trp Ile Pro Gln Glu Gln Gly Ser Pro Gln Ser Gly Ser Pro His Gly 325 330 335 Pro Leu His His Leu Ser Thr Ser Val Ala Ala Val Ser Ser Ala Ala 340 345 350 Thr Ala Ser Cys Glu Glu Glu Asp Gly Arg Ser Glu Ser Tyr Gly Trp 355 360 365 Lys <210> 2 <211> 365 <212> PRT <213> Zea mays <400> 2 Met Gly Leu Asp Val Met Glu Ile Gly Met Gly Ala Asp Leu Ser Leu 1 5 10 15 Asp Leu Arg His Phe Ala Ser Lys Ala Val Arg Gln Ser Lys Asp Asp 20 25 30 Thr Pro Ala Pro Asp Met Asp Ala Cys Ile Arg Arg Leu Glu Glu Glu 35 40 45 Arg Gly Lys Ile Glu Met Phe Lys Arg Asp Leu Pro Leu Cys Ala Arg 50 55 60 Leu Leu Ala Asp Val Ile Asp Val Met Lys Glu Glu Ala Gly Lys Lys 65 70 75 80 Lys Thr Thr Thr Arg Arg Ser Asp Arg Arg Leu Ala Ser Ala Ala Ala 85 90 95 Asp Glu Glu Glu Glu Glu Glu Asp Gly Ala Thr Ala Asp Lys Ser Lys 100 105 110 Trp Met Ser Thr Ala Gln Leu Trp Thr Gly Asp Ser Gly Arg Glu Asp 115 120 125 Ala Glu Ser Glu Lys Gln Asp Lys Gly Trp Cys Ser Pro Glu Ala Arg 130 135 140 Ser Arg Gly Ala Leu Leu Pro Phe Lys Ala Glu Val Gly Ser Gly Ala 145 150 155 160 Pro Ala Phe Ala Pro Leu Cys Leu Arg Thr Asp Asp Lys Ala Ala Ala 165 170 175 Ala Arg Val Gly Val Pro Asp Leu Ser Ser Leu Leu Ser Ser Pro Ala 180 185 190 Thr Met Pro Pro Ala Asp Ala Gly Ala Glu Glu Ser Arg Arg Gln Val 195 200 205 Val Gly Phe Ala Gln Ala Ala Ala Arg Ala Ala Ala Met Ala Pro Ser 210 215 220 Ala Pro Ala Leu Gly Leu Gln Ser Gln Gln Gln Gln Gln Gln Ala Arg 225 230 235 240 Lys Ala Arg Arg Cys Trp Ser Thr Glu Leu His Arg Lys Phe Val Ala 245 250 255 Ala Leu Asp Gln Leu Gly Gly Pro Gln Val Ala Thr Pro Lys Gln Ile 260 265 270 Arg Glu Leu Met Lys Val Asp Gly Leu Thr Asn Asp Glu Val Lys Ser 275 280 285 His Leu Gln Lys Tyr Arg Leu His Asn Arg Arg Ala Pro Gly Ser Gly 290 295 300 Val Val Arg Gln Pro Ile Val Leu Val Gly Gly Leu Trp Ile Pro Gln 305 310 315 320<00005'06>Glu Gln Gly Ser Pro Gln Ser Gly Ser Pro His Gly Pro Leu His His s325 330 335 Leu Ser Thr Ser Val Ala Ala Val Ser Ser Ala Ala Thr Ala Ser Cys 340 345 350 Glu Glu Glu Asp Gly Arg Ser Glu Ser Tyr Gly Trp Gln 355 360 365 <210> 3 <211> 367 <212> PRT <213> Maize <400> 3 Met Gly Leu Asp Val Met Glu Ile Gly Met Gly Ala Asp Leu Ser Leu 1 5 10 15 Asp Leu Arg His Phe Ala Ser Lys Ala Val Arg Gln Ser Lys Asp Asp 20 25 30 Thr Pro Ala Pro Asp Met Asp Ala Cys Ile Arg Arg Leu Glu Glu Glu 35 40 45 Arg Gly Lys Ile Glu Met Phe Lys Arg Asp Leu Pro Leu Cys Ala Arg 50 55 60 Leu Leu Ala Asp Val Ile Asp Val Met Lys Glu Glu Ala Gly Lys Lys 65 70 75 80 Thr Thr Thr Thr Arg Arg Ser Asp Arg Arg Leu Ala Ser Ala Ala Ala 85 90 95 Asp Glu Glu Glu Glu Glu Glu Asp Gly Ala Thr Ala Asp Lys Ser Lys 100 105 110 Trp Met Ser Thr Ala Gln Leu Trp Thr Gly Asp Ser Gly Arg Glu Asp 115 120 125 Ala Glu Ser Glu Lys Gln Asp Lys Gly Arg Cys Ser Pro Glu Ala Arg 130 135 140 Ser Arg Gly Ala Leu Leu Arg Phe Lys Ala Asp Val Gly Ser Gly Ala 145 150 155 160 Pro Ala Phe Ala Pro Leu Cys Leu Arg Thr Asp Asp Lys Ala Ala Ala 165 170 175 Ala Arg Val Gly Val Pro Asp Leu Ser Ser Leu Leu Ser Pro Pro Ala 180 185 190 Thr Met Pro Pro Ala Asp Ala Gly Ala Glu Glu Ser Arg Arg Gln Val 195 200 205 Val Gly Phe Ala Gln Ala Ala Ala Arg Ala Ala Ala Met Ala Pro Ser 210 215 220 Ala His Ala Leu Gly His Gln Ser Gln Ser Gln Gln Gln Gln Gln Gln 225 230 235 240 Ala Arg Lys Ala Arg Arg Cys Trp Ser Thr Glu Leu His Arg Lys Phe 245 250 255 Val Ala Ala Leu Asp Gln Leu Gly Gly Pro Gln Val Ala Thr Pro Lys 260 265 270 Gln Ile Arg Glu Leu Met Lys Val Asp Gly Leu Thr Asn Asp Glu Val 275 280 285 Lys Ser His Leu Gln Lys Tyr Arg Leu His Asn Arg Arg Ala Pro Gly 290 295 300 Ser Gly Val Val Arg Gln Pro Ile Val Leu Val Gly Gly Leu Trp Ile 305 310 315 320 Pro Gln Glu Gln Gly Ser Pro Gln Ser Gly Ser Pro His Gly Pro Leu 325 330 335 His His Leu Ser Thr Ser Val Ala Ala Val Ser Ser Ala Ala Thr Ala 340 345 350 Ser Cys Glu Glu Glu Asp Gly Arg Ser Glu Ser Tyr Gly Trp Lys 355 360 365 <210> 4 <211> 5006 <212> DNA <213> Zea mays <400> 4 cgttcacgcg atttttcaag tgaaagcgag accgaaaacc agcaatgggc tgtaggaatg 60 cattaggata tttttaccaa atattagcca aacgtttttg ttattatgca taatagtagg 120 catagcacta aatttacaat agactacaaa tataggggct ttattacaaa atattctttc 180 ggacgtgaaa actcgtcatt gttgattcgg agattctacc actccatctc cagttttttc 240 cctcgcctgc tcagctcccc tataaatgga gctcgccttc cgcggcctcc ctccgttccc 300 atccgccgcc cgcgcacttc ttccttcggg cacacaggac accaccgtcg acggattcat 360 cgcgacgatg gggctcgacg tcatggagat cgggatgggc gccgatttga gcctggatct 420 gaggcacttc gcctccaagg ccgtgaggca gagcaaggac gacacgccgg cgccggacat 480 ggacgcatgc atccgccgcc tcgaggagga gcggggtaag atcgagatgt tcaagcggga 540 cctcccgctc tgcgcgcgcc tcctcgccga cggtgagcgc acctacctct tctcctctct 600 ctctgtctct ctctcttttt atttttccca cctgtgattc atttgggata ccttttgctt 660 ctttccattt tggggagcgg ttttttttac gcggcgatgc ggtggcgtgt gcgcagtaat 720 tgatgtcatg aaggaggagg cggggaagaa gaagacgacg acaaggagga gtgaccgcag 780 gctggcgtct gcggcagctg atgaggagga ggaggaggag gacggcgcca ccgcggacaa 840 gagcaagtgg atgagcacgg cgcagctctg gacgggcgat tccgggcggg aggacgcgga 900 atcagaggta cggcacgatt cgatcgctgg tgcagctgct tgaatgctca gtcagcacag 960 gatctggagg gtgctgtcgg gtgctcgatt cgtcggcagg cctaaaagtt tggagctttg 1020 cgatcgcaga agcaagacaa ggggtggtgc tcgccggagg ccaggtcccg cggcgctctc 1080 ttaccgttca aggctgaagt gggctctggc gcgccggcgt tcgcgccgct ctgcttgaga 1140 acggacgaca aggctgcggc tgcgcgcgtc ggggtgccgg atctgtcgtc cttgctgtcg 1200 tcgccggcga ccatgcctcc tgcggacgcc ggcgccgagg agagccgtcg ccaggttgtg 1260 ggatttgcgc aagctgcggc cagggcggct gccatggcgc cgtctgcccc tgcgcttggg 1320 ctccagtcgc agcagcagca gcagcaggca aggaaggctc ggcgttgctg gtcgacggag 1380 ctgcatcgca agttcgtcgc cgccttggat cagctcggtg gcccccaagg tgagccttgc 1440 cttgttcttc ggatgccagt tcaccagaat ctcttgccag ttttgagcca ccaacacgtt 1500 caatcttacc tagttgctag ctgccttcca tattagaatc ataaaattgg gatcaatgag 1560 tctatgccat gactgcagtt gccacgccga agcaaatcag ggagctgatg aaggtggatg 1620 ggctgacaaa cgacgaagtg aaaagccatc ttcaggttag cgatccagca gcagctcact 1680 ccccttgcca ttccattcat ccatctcatc tcaggaagtc acgagtatct gttgttgtga 1740 tggttgctga aatggattct ccgatttcga tgtctcttca gaaataccgg ctgcacaacc 1800 ggagggcgcc tggatccggc gtggtgcgcc agccgatcgt gctcgtggga gggctgtgga 1860 ttccccagga gcaaggcagc cctcagtctg gatctcccca cggccccctc caccacctgt 1920 ccacctcggt ggccgccgtc tcgtccgccg ccaccgccag ctgcgaggag gaagacggcc 1980 ggtccgagag ctatggctgg caatgatgtc tggctgctgc tgctgctgca ccaccaatgt 2040 gtgttcactg ttcagagagg ggaggttct tggcatggtg gggatcgcca tgggccatgg 2100 cggaggccac cagttgcagc ttcaggaatc gggaggggaa ttgagtgtag tgtagctgtc 2160 tgtacacata catacataca tacagtgaga tgggatgaga tgagagcggg ccttgagcgc 2220 tcgagatcag aactgatggt gcttcgtcgt cgggtttgta catcccaaag agaaagagat 2280 actagctaca gttttgcggc ttgttaatcc atgctctggg ggcagagcta cagttttcgc 2340 cccgagagag ttcacccata cccgttgttg tcgattagac gattaccatc ttcgccttct 2400 tgttgccgtt gaacaaaatg ttgcttccgc tgttcgtctc ggaacgaaca gtccggttga 2460 aaagttgaat cgttgcagga gtacatgcta ctcaggctgt aatgtggttg gtaagggtgt 2520 ttgaatgaac tagacctaat agttagtgac taaatagt tggatacatc taaacatcct 2580 ataatcctat agtttaacta ttagatattt gttatctcgc taattttata agtaattttt 2640 agccaactaa ctattagttc taatgcattc gaacgctcac tctgcagctt tccagcgcg 2700 tatcgttcag gtctatctaa ctgaaagagc agcaagaag aaagagatct cataagaaa 2760 agaacccgat tccaccattg aaaccaac caaggtt gctcctgctc ctctgctgtt 2820 caccatcatc aacagcacgt aaaaaaaatc tacttaccg tctaccacgta 2880 gtgttttggat aaagcacag aagccggctt ctctcttttt ttgcagaga taatatagat 2940 attcagaga agaaaaaga attatctggg cctactgaa actgagctga cggagcacga 3000 gcacggaagc catgcttgtt gtatacataa cataagccgg ggggaggata tgctcgaggc 3060 attctctct tcctcctccg tccactg gctcggtcca tcgttagcg tcgtcg 3120 ccttgatcag cattgttaat actactagct cgctgctgag tgctgacaat gcgaaacagt 3180 ttcttggcag gattccaact cgagctcgcc gtcgctgtcg ctgtcgctgg accgtaggaa 3240 cgtgccggtc ccctccctgc atggcgggcc aagagccac ccagatcaga ggacggatcc 3300 ccgtgaaatc cccctcttg ttctttaatt accgcaggc ggaggaagg cggcagtgca 3360 cagcgacaga gagacgaga ctttggaatc gtccttggtt ggatggatgg acggacgaac 3420 gaggcggggc cgcgagctct gaatattcgc cgccgtcgat gcatcggcgg cctgcctgtc 3480 gctgtcgacg gagagggtgg tactggtgtg cgcaaccgga caacgcaatg ttcaggcctg 3540 aagaatcgga atcggaatat tatattccgt ctggtgtggt tgtcactttc ttttctttgt 3600 gtgtgttttt ttgttgtttg ttgttgttgt gtagataaat actatgggga agaatggagg 3660 gggatatgag gatatcctcg ttgattctgc ttgagaaact agggcgtatt atatgataca 3720 tacatttgga attctcactc tcggccgacc ccgccggcga cctcagcccg acgtggatat 3780 ataaaaaaa agatgattaa agctttgtaa gataagacta gtctgcactt tctagtagat 3840 ttagaccata ttttgaaacg tctgaaacta ataattaaca gataaaacta gctaagagag 3900 aacgaatcag ctaatagatt agctaattgc tggttacatc tctcaaatag ctattagtta 3960 ttagttaatt taatctagct aaaatcaaat acaacaactt actctttatg tacaaaatta 4020 aaatttgttt tagtttttta ttggattcat ataataattt gtgtttatgt ttttttatat 4080 gtttctaaat ttattatata aaaactaaga gataaaatga ataataattt ttggacggaa 4140 agaatattag ctccctacag ttttgggaca ggtcgcaact tgtctgacta gttaagttttc 4200 tatagcgcgg tagtagtcta gtagaagata gtactctctt tgtttctttt tagttattat 4260 tggataattt aatttgtaa tattcagcga caactaaaac gaaacgtaag agagggtaga 4320 taactttgga gacttgagtc gtcgtgaatg ggatgggact tgtcggagcc tcggcgcagc 4380 gtattattg ttgacagggc cgtgagagcc tggtccacat tttgttggcc catttaggtg 4440 agctgtctac agattgggcc gagcaagtaa gggtatagga agccgaaatg tgcccattta 4500 accaatcatc acggtttgag tcgacattc acgattctgc aaccacagta cttatttat 4560 ttggttgaaa acacaaggtt aattaatact aacagtagcg acaatgatga tgctccttca 4620 cgcttccttg ccatatcata aaaaaacagt aaaaaggtaa aagaaaaagg ttaatgccta 4680 cctatagctt ttagcttgca gcgcgccctc tctctcttct ctctgtatat atgccgtgat 4740 cgccggcaca catcgcggcg tgttgcatt ccgatcggcg gccgcgaaaa aggaaaaata 4800 aagaagtgaa aatagagga aagcagaaag aataaaagag ggctaaaaga aaaaggcatg 4860 tcgccgtatg ctggcgcctt gatagtcagg ggctcagggc gtcaggcaga catgcttgta 4920 gtagttagta tatagagtcc ggccgacacg gttcagcggc cacatgcatg cagcgacagg 4980 ctaatcaaag cccacaacag agaccg 5006 <210> 5 <211> 5256 <212> DNA <213> Zea mays <400> 5 cgttcacgcg atttttcaag tgaaagccaa atattagcca aacgtttttg ttattatgca 60 taatagtagg catagcacta aatttacaat agactacaaa tataggggct ttcgtacaaa 120 ataatctttc ggacgtgaaa actcgtcatt gttgattcgg agattctacc actccatctc 180 cagttttttt ttccctcgcc tgctcggctc ccctataaat ggagctcacc ttccgcggcc 240<了 tccctccgtt cccatccgcc gcccgcgcac ttcttccttc gggcacacag gacaccaccg 300 tcgacggatt catcgcgacg atggggctcg acgtcatgga gatcgggatg ggcgccgatt 360 tgagcctgga tctgaggcac ttcgcctcca aggccgtgag gcagagcaag gacgacacgc 420 cggcgccgga catggacgca tgcatccgcc gcctcgagga ggagcggggt aagatcgaga 480 It should be noted that there seems to be an error in the tag "<了 " in the original text. It might be a misspelling. If this is an important part, it should be corrected for a more accurate translation.tgttcaagcg ggacctcccg ctctgcgcgc gcctcctcgc cgacggtgag cgcacctacc 540 tcttctctct ctgtctctct ctctttttta tttttcccac ctgtgattca tttgggatac 600. cttctgcttc tttccatttt ggggagcggg tttttttatg cggcgatgtg gtggcgtgtg 660 cgcagtaatt gatgtcatga aggaggaggc ggggaagaag acgacgacca cgaggaggag 720 tgatcgcagg ctggcgtctg cggcagctga tgaggaggag gaggagg acggcgccac 780 cgcggacaag agcaagtgga tgagcacggc gcagctctgg acggcgatt ccggggcggga 840 ggacgcgga tcagaggtac ggcacgattc gatcgctggt gcagctgctt gaatgcccag tcagcacagg atctgggggg tgctgtcggg tgctcgattc gtcggcaggc ctaaaagttt 960 tggagctttg cgatcgcaga agcaagacaa ggggcggtgc tcgccggagg ccaggtcccg cggcgctctc ttacggttca aggctgatgt gggctctggc gcgccggcgt tcgcgccgct 1080 ctgcttgaga acggacgaca aggctgcggc tgcgcgcgtc ggggtgccgg atctgtcgtc 1140 cttgctgtcg ccgccggcga ccatgcctcc tgcggacgcc ggcgccgagg agagccgtcg ccaggttgtg ggatttgcgc aagctgcggc cagggcggct gccatggcgc cgtctgccca 1260 tgcgcttggg caccagtcgc agtcgcagca gcagcagcag caggcaagga aggctcggcg 1320 ttgctggtcg acggagctgc atcgcaagtt cgtcgccgcc ttggatcagc tcggtggccc 1380 ccaaggtgag ccttgccttg ttcttcggat gccagttcac cagaatttct tgccagtttt 1440 gggccaccaa cacacacgtt caatcttacc tagttgctag ctgccttcca tattatatta 1500 gaaacactga gttcattcat gctacgccat gcctgcagtt gccacgccga agcaaatcag 1560 ggagctgatg aaggtggatg ggctgacaaa cgacgaagtg aaaagccatc ttcaggttag 1620 cgatccagca gcagctcact ccccttgaca ttccattcat ccatctcatc tcaggaagtc 1680 acgaatatct gttgttgtga tggttgctga aatggattct ctgatttcga tgtttgttca 1740 gaaataccgg ctgcacaacc gcagggcgcc tggatccggc gtggtgcgcc agccgatcgt 1800 gctcgtggga gggctgtgga ttccccagga gcaaggcagc cctcagtctg gatctcccca 1860 cggccctctc caccacttgt ccacctcggt ggccgccgtc tcgtccgccg ccaccgccag 1920 ctgcgaggag gaagacggcc ggtccgagag ctatggctgg aaatgatgaa gaggctgctg 1980 ctgctgctgc accaccaatg tgtgttcact gttagagag gggaggtttc ttggcatggt 2040 ggggatcgcc atgggccatg gcggaggcca ccagttgcag cttcaggaat cgggagggga 2100 attgagtgta gtgtagctgt ctgtacacat acatacatac atacattgag atgggatgag 2160 atgagagcgg gccttgagcg ctcgagatca gaactgatgg tgcttcgtcg tcgggtttgt 2220 acatcccaaa gagaaagaga aagagatact agctacagtt ttgcggcttg ctaatccatg 2280 cctgggggca gagctacagt ttcgccccg agagagttca cccatcaccc atacccgttg 2340 ttgtcgatta ccatcttcgc cttcttgttg ccgttgaaca aaatgttgct ttcgctgttc 2400 gtctcggaac gaacagtccg gttgaaaagt tgaatcgttg caggagtaca tgctactgag 2460 tctgtaatgt ggttggtaag ggtgtttgaa tgaactagac ctaatagtta gtgactaaaa 2520 ttagaatcat atagtttaac tattagatat ttgttatctc gctaatttta tagtaattt 2580 ttagccaact aactattagt tcgaatgcat tcgaacactc actctgcagc tttccagtcg 2640 cgtatcgtta aggtctatct aactgaaaga gcagcaaga tgaagagat ctcataaga 2700 aaagaacccg attccaccat tgacaacca accagaggg tgctcctgc tcctctgctg 2760 ttcaccatca tcacagcac gtaaaaaaaa aatcttagc tctctactgt accgtctacc 2820 agtagtgttt ggataaagca acagacccg gttctctctc ttttacag agataatata 2880 gatattcaga agagaaga aaaggataa ttatctggggc ctactgaaa ctgagctgac 2940 ggagcacgag cacggaagcc atgcttgttg tatacataac ataagccggg gggaggatat 3000 gctcgaggca ttctctctt cctcatccgt cagtcactgg ctcggtccat tcgttagcgt 3060 ctcaccagtc cttgatcagc attgttaata ctactagctc gctgctgagt gctgactatg 3120 cgaaacagtt tcttggcagg attccacaa ctcgagctcg ccgtcgccgt cgctgtcgct 3180 ggaccgtacg aacgtgccgg tccctccct gcatggcgga ccaagagcc acccagatca 3240 ggacggatcc ccgtgaaatc ccccttg ttctattt actcgcaggc ggaggaaagg 3300 cggcagtgca cagcgacaga gagacgaga ctttggaatc gtccttgggt gcatggatgg 3360 acggacggac gaacgagagg gggggccgcg agctctgaat attcgccgcc gtcgatgcat 3420 cggcggcctg cctgtcgctc tcgacggaga gggtggtact ggtgtgcgca accggacaac 3480 gcaatgttca ggccagaaga atcggaatcg gaatatcata ttccgtctgg tgtggtggtc 3540 actttcttt ctttgtgtgt gtgtttttt gttgttgttg tgtagataaa tactatgggg 3600 aagaatggag gggatatgag gatatcctcg ttgattctgc ttgagaaact agggcgtatt 3660 atatgataca tttggaattc tcactctcgg ctgggccgac cccgccggcg acctcagccc 3720 gacgtggata tatataaaaa gatgattaaa gctttgtaag ataagactag tctgcacttt 3780 ctagtagatt tagaccatat tttcaaacgt ctaaaactaa taatgaacag ataaactag 3840 ctaagagaga acgaatcagc taatagatta gctaattgtt agttacattt ctcaaatagc 3900 tattagttgt tagttaattt aatctagcta aaatcaacta caacaactta ctctttatgt 3960 acaaaattaa aatttgtttt agttttttat tggattcata taataatttg tgtttatgtt 4020 tttttatatg tttctaaatt tattatataa aaactaagag ataaaatgaa taataatttt 4080 tggacggaaa gaatattagc tccctacagt tttgggacag gtcgcaactt gtctgactag ttagtttct atagcgcggt agtagtctag tag tactctcttt gtttcttttt 4200. agttattatt ggataattta attttgtaat attcagcgac aactaaacg aaacgtaaga gagggtag aactttggag acttgagtcg tcgtgaatgg gatgggactt gtcggagcct cggcgcagcg tattatttgt tgacagggcc gtgagagcct ggtccacatt ttgttggccc 4380. atttaggtga gctgtctaca gattgggccg agcaagtaag ggtataggaa gccgaaatgt gcccatttaa ccaatcatca cggtttgagt cgacattcca cgattctgca accacagtac tttatttatt tggttgaaaa cacaaggtta attack acagtagcga caatgatgat gctccttcac gcttccttgc catatcata aaaacagtaa aaaggtaaaa gaaaaggtt aatgcctacc tatagcttgc agcgcgccct ctctctcttc tctctgtata tatgccgtga 4680 tcgccggcac acatcgcggc gtgtttccat tccgatcggc ggcggcgaaa aagggaaaaa 4800. 4800. 4800. 4800. 4800. 4800. 4800. 4800. 4800 tgtcgccgta tgctggcgcc ttgatagtca ggcagacatg ctttgcagta gtagtagtat 4860 atagggtgtg tttggtttga cttttgactc tggcttttac cccctaaaag ctaaaagcca 4920 aaccaaaggg ctggatttag gaagcagctt tttctaaaag ccgactttct tgcagtgcaa 4980 aactgaaagc acctctagac ctgcttttag ctgcttttag atggaactgt gaaaatatat 5040 atggaaaaac atttagcgac ttttagtggt ttccaccaaa cactttttag ctttttaaca 5100 gctcgcagcc cacagcagct tttctcacag ctcacagccc acagcagctt ttttcacagc 5160 cacagtccaa ccaaacagac catagagtcc ggccgacacg gttcagcggc cacatgcatg 5220 cagcgacagt ctaatcaaag cccacaacag agaccg 5256 <210> 6 <211> 2084 <212> DNA <213> Zea mays <400> 6 gttcccatcc gccgcccgcg cacttcttcc ttcgggcaca caggacacca ccgtcgacgg 60 attcatcgcg acgatggggc tcgacgtcat ggagatcggg atgggcgccg atttgagcct 120 ggatctgagg cacttcgcct ccaaggccgt gaggcagagc aaggacgaca cgccggcgcc 180 ggacatggac gcatgcatcc gccgcctcga ggaggagcgg ggtaagatcg agatgttcaa 240 gcgggacctc ccgctctgcg cgcgcctcct cgccgacggt gagcgcacct acctcttctc 300 ctctcttctg tctctctctt ttttattttt cccacctgtg attcatttgg gataccttct 360 gcttctttcc attttgggga gcggtttttt ttatgcggcg atgcggtggc gtgtgcgcag 420 taattgatgt catgaaggag gaggcgggga agaagaagac gacgacgagg aggaggagtg 480 atcgcaggct ggcgtctgcg gcagctgatg atgaggagga ggaggcggac ggcgccaccg 540 cggacaagag caagtggatg agcacggcgc agctctggac gggcgattcc gggcgggagg 600 acgcggaatc agaggtacgg cacgattcga tcgctggtgc agctgcttga atgctcagtc 660 agcacaggat ctgtgggggt gctgtcgggt gctcgattcg tcggtgggcc taaaagtttt 720 ggagctttgc gatcgcagaa gcaagacaag gggcggtgct cgccggaggc caggtcccgc 780 ggcgctctct taccgttcaa ggctgatgtg ggctctggcg cgccggcgtt cgcgccgctc 840 ttcttgagaa cggacgacaa ggctgcggct gcgcgcgtcg gggtgccgga tctgtcgtcc 900 ttgctgtcgc cgccggcgac catgcctcct gcggacgccg gcgccgagga gagccgtcgc 960 caggttgtgg gatttgcgca agctgcggcc agggcggctg ccatggcgcc gtctgcccct 1020 gcgcttgggc tccagtcgca gcagcagcag cagcagcagc agcaggcaag gaaggctcgg 1080 cgttgctggt cgacggagct gcatcgcaag ttcgtcgccg ccttggatca gctcggtggc 1140 ccccaaggtg agccttgcct tgttcttcgg atgccagttc accagaattt cttgccagtt 1200 ttgggccacc aacacacacg tccaatctta cctagttgct agctgccttc catattatat 1260 tagaaagaaa cattgagttc attcatgcta cgccatgcct gcagttgcca cgccgaagca 1320 aatcagggag ctgatgaagg tggatgggct gacaaacgac gaagtgaaaa gccatcttca 1380 ggttagcgat ccagcagcag ctcagtccac ttggcattcc attcatccat ctcaggaagt 1440 cacgaatatc tgttgttttg atggttgctg aaatggattc tctaattccg atgtttattc 1500 agaaataccg gctgcacaac cgcagggcgc ctggatccgg cgtggtgcgc cagccgatcg 1560 tgctcgtggg agggctgtgg attccccagg agcaaggcag ccctcagtct ggatctcccc 1620 acggccccct ccaccacctg tccacctcgg tggccgccgt ctcgtccgcc gccaccgcca 1680 gctgcgagga ggaagacggc cggtccgaga gctatggctg gaaatgatga agaggctgct 1740 gctgctgctg ctgcgccacc aatgtgtgtt cactgtttag agaggggagg gaggtttctt 1800 ggcatggtgg ggatcgccat gggccatggc ggaggccacc agttgcagct tcaggaatcg 1860 ggaggggaat tgagtgtagt gtagctgtct gtacacatac atacatacat acagtgagat 1920 gggatgagat gagagcgggc cttgagcgct cgagatcaga actgatggtg cttcgtcgtc 1980 gggtttgtac atcccaaaga gaaagagata ctagctacag ttttgcggct tgttaatcca 2040 tgctctgggg gcagagctac agttttcgcc ccgagagagt tcac 2084 <210> 7 <211> 1540 <212> DNA <213> Zea mays <400> 7 gttcccatcc gccgcccgcg cacttcttcc ttcgggcaca caggacacca ccgtcgacgg 60 attcatcgcg acgatggggc tcgacgtcat ggagatcggg atgggcgccg atttgagcct 120 ggatctgagg cacttcgcct ccaaggccgt gaggcagagc aaggacgaca cgccggcgcc 180 ggacatggac gcatgcatcc gccgcctcga ggaggagcgg ggtaagatcg agatgttcaa 240 gcgggacctc ccgctctgcg cgcgcctcct cgccgacgta attgatgtca tgaaggagga 300 ggcggggaag aagaagacga cgacgaggag gaggagtgat cgcaggctgg cgtctgcggc 360 agctgatgat gaggaggagg aggcggacgg cgccaccgcg gacaagagca agtggatgag 420 cacggcgcag ctctggacgg gcgattccgg gcgggaggac gcggaatcag agaagcaaga 480 caaggggcgg tgctcgccgg aggccaggtc ccgcggcgct ctcttaccgt tcaaggctga 540 tgtgggctct ggcgcgccgg cgtcgcgcc gctcttttg agaacggacg acaaggctgc 600 ggctgcgcgc gtcggggtgc cggatctgtc gtccttgctg tcgccgcgg cgaccatgcc 660 tcctgcggac gccggcgccg aggagagccg tcgccaggtt gtgggatttg cgcaagctgc 720 ggccagggcg gctgccatgg cgccgtctgc cctgcgctt gggctccagt cgcagcagca 780 gcagcagcag cagcagcagg caaggaaggc tcggcgttgc tggtcgacgg agctgcatcg 840 caagttcgtc gccgccttgg atcagctcgg tggcccccaa gttgccacgc cgaagcaaat 900 cagggagctg atgaaggtgg atgggctgac aaacgacgaa gtgaaaagcc atcttcagaa 960 ataccggctg cacaaccgca gggcgcctgg atccggcgtg gtgcgccagc cgatcgtgct 1020 cgtgggaggg ctgtggattc cccaggagca aggcagccct cagtctggat ctccccacgg 1080 ccccctccac cacctgtcca cctcggtggc cgccgtctcg tccgccgcca ccgccagctg 1140 cgaggaggaa gacggccggt ccgagagcta tggctggaaa tgatgaagag gctgctgctg 1200 ctgctgctgc gccaccaatg tgtgttcact gtttagagag gggagggagg tttcttggca 1260 tggtggggat cgccatgggc catggcggag gccaccagtt gcagcttcag gaatcgggag 1320 gggaattgag tgtagtgtag ctgtctgtac acatacatac atacatacag tgagatggga 1380 tgagatgaga gcgggccttg agcgctcgag atcagaactg atggtgcttc gtcgtcgggt 1440 ttgtacatcc caaagagaaa gagatactag ctacagtttt gcggcttgtt aatccatgct 1500 ctgggggcag agctacagtt ttcgccccga gagagttcac 1540 <210> 8 <211> 1,098 <212> DNA <213> Zea mays <400> 8 atggggctcg acgtcatgga gatcgggatg ggcgccgatt tgagcctgga tctgaggcac 60 ttcgcctcca aggccgtgag gcagagcaag gacgacacgc cggcgccgga catggacgca 120 tgcatccgcc gcctcgagga ggagcggggt aagatcgaga tgttcaagcg ggacctcccg 180 ctctgcgcgc gcctcctcgc cgacgtaatt gatgtcatga aggaggaggc ggggaagaag 240 aagacgacga caaggaggag tgaccgcagg ctggcgtctg cggcagctga tgaggaggag 300 gaggaggagg acggcgccac cgcggacaag agcaagtgga tgagcacggc gcagctctgg 360 acgggcgatt ccgggcggga ggacgcggaa tcagagaagc aagacaaggg gtggtgctcg 420 ccggaggcca ggtcccgcgg cgctctctta ccgttcaagg ctgaagtggg ctctggcgcg 480 ccggcgttcg cgccgctctg cttgagaacg gacgacaagg ctgcggctgc gcgcgtcggg 540 gtgccggatc tgtcgtcctt gctgtcgtcg ccggcgacca tgcctcctgc ggacgccggc 600 gccgaggaga gccgtcgcca ggttgtggga tttgcgcaag ctgcggccag ggcggctgcc 660 atggcgccgt ctgcccctgc gcttgggctc cagtcgcagc agcagcagca gcaggcaagg 720 aaggctcggc gttgctggtc gacggagctg catcgcaagt tcgtcgccgc cttggatcag 780 ctcggtggcc cccaagttgc cacgccgaag caaatcaggg agctgatgaa ggtggatggg 840 ctgacaaacg acgaagtgaa aagccatctt cagaaatacc ggctgcacaa ccggagggcg 900 cctggatccg gcgtggtgcg ccagccgatc gtgctcgtgg gagggctgtg gattccccag 960 gagcaaggca gccctcagtc tggatctccc cacggccccc tccaccacct gtccacctcg 1020 gtggccgccg tctcgtccgc cgccaccgcc agctgcgagg aggaagacgg ccggtccgag 1080 agctatggct ggcaatga 1098 <210> 9 <211> 1104 [[ID=)18]]<212> DNA <213> Zea mays <400> 9 atggggctcg acgtcatgga gatcgggatg ggcgccgatt tgagcctgga tctgaggcac 60 ttcgcctcca aggccgtgag gcagagcaag gacgacacgc cggcgccgga catggacgca 120 tgcatccgcc gcctcgagga ggagcggggt aagatcgaga tgttcaagcg ggacctcccg 180 ctctgcgcgc gcctcctcgc cgacgtaatt gatgtcatga aggaggaggc ggggaagaag 240 Note: There seems to be a formatting issue in the original text where the closing parenthesis in line 18's "DNA" is missing in the original Chinese you provided. I've added it in the translation for correct representation. Also, "玉蜀黍" is translated as "Zea mays" which is the scientific name for corn.acgacgacca cgaggaggag tgatcgcagg ctggcgtctg cggcagctga tgaggaggag 300 gaggaggagg acggcgccac cgcggacaag agcaagtgga tgagcacggc gcagctctgg 360 acgggcgatt ccgggcggga ggacgcggaa tcagagaagc aagacaaggg gcggtgctcg 420 ccggaggcca ggtcccgcgg cgctctctta cggttcaagg ctgatgtggg ctctggcgcg 480 ccggcgttcg cgccgctctg cttgagaacg gacgacaagg ctgcggctgc gcgcgtcggg 540 gtgccggatc tgtcgtcctt gctgtcgccg ccggcgacca tgcctcctgc ggacgccggc 600 gccgaggaga gccgtcgcca ggttgtggga tttgcgcaag ctgcggccag ggcggctgcc 660 atggcgccgt ctgcccatgc gcttgggcac cagtcgcagt cgcagcagca gcagcagcag 720 gcaaggaagg ctcggcgttg ctggtcgacg gagctgcatc gcaagttcgt cgccgccttg 780 gatcagctcg gtggccccca agttgccacg ccgaagcaaa tcagggagct gatgaaggtg 840 gatgggctga caaacgacga agtgaaaagc catcttcaga aataccggct gcacaaccgc 900 agggcgcctg gatccggcgt ggtgcgccag ccgatcgtgc tcgtgggagg gctgtggatt 960 ccccaggagc aaggcagccc tcagtctgga tctccccacg gccctctcca ccacttgtcc 1020 acctcggtgg ccgccgtctc gtccgccgcc accgccagct gcgaggagga agacggccgg 1080 tccgagagct atggctggaa atga 1104 <210> 10 <211> 349 <212> DNA <213> Zea mays <400> 10 cagacatgct ttgcagtagt agtagtatat agggtgtgtt tggtttgact tttgactctg 60 gcttttaccc cctaaaagct aaaagccaaa ccaaagggct ggatttagga agcagctttt 120 tctaaaagcc gactttcttg cagtgcaaaa ctgaaagcac ctctagacct gcttttagct 180 gcttttagat ggaactgtga aaatatatat ggaaaaacat ttagcgactt ttagtggttt 240 ccaccaaaca ctttttagct ttttaacagc tcgcagccca cagcagcttt tctcacagct 300 cacagcccac agcagctttt ttcacagcca cagtccaacc aaacagacc 349 <210> 11 <211> 20 <212> DNA <213> Zea mays <400> 11 cggtctctgt tgtgggcttt 20 <210> 12 <211> 20 <212> DNA Note: There is a possible error in the original text where "0000875" should likely be "0000875". This has been noted in the translation as "0000875" is not a standard format. Also, "Zea mays" is the scientific name for corn (maize), which is the translation for "玉蜀黍". <213> Maize <400> 12 atgctggcgc cttgatagtc 20 <210> 13 <211> 1048 <212> DNA <213> Maize <400> 13 aattatctgg gcctaactga aactgagctg acggagcacg agcacggaag ccatgcttgt 60 tgtatacata acataagccg gggggaggat atgctcgagg cattctcttc ttcctcctcc 120 gtcagtcact ggctcggtcc attcgttagc gtctcaccag tccttgatca gcattgttaa 180 tactactagc tcgctgctga gtgctgacaa tgcgaaacag tttcttggca ggattccaac 240 tcgagctcgc cgtcgctgtc gctgtcgctg gaccgtagga acgtgccggt cccctccctg 300 catggcgggc caaagagcca cccagatcag aggacggatc cccgtgaaat cccccctctt 360 gttctttaat tactcgcagg cggaggaaag gcggcagtgc acagcgacag agagacgaag 420 actttggaat cgtccttggg tggatggatg gacggacgaa cgaggcgggg ccgcgagctc 480 tgaatattcg ccgccgtcga tgcatcggcg gcctgcctgt cgctgtcgac ggagagggtg 540 gtactggtgt gcgcaaccgg acaacgcaat gttcaggcct gaagaatcgg aatcggaata 600 ttatattccg tctggtgtgg ttgtcacttt cttttctttg tgtgtgtttt tttgttgttt 660 gttgttgttg tgtagataaa tactatgggg aagaatggag ggggatatga ggatatcctc 720 gttgattctg cttgagaaac tagggcgtat tatatgatac atacatttgg aattctcact 780 ctcggccgac cccgccggcg acctcagccc gacgtggata tataaaaaaa aagatgatta 840 aagctttgta agataagact agtctgcact ttctagtaga tttagaccat attttgaaac 900 gtctgaaact aataattaac agataaaact agctaagaga gaacgaatca gctaatagat 960 tagctaattg ctggttacat ctctcaaata gctattagtt attagttaat ttaatctagc 1020 taaaatcaaa tacaacaact tactcttt 1048 <210> 14 <211> 1048 <212> DNA <213> Zea mays <400> 14 aattatctgg gcctaactga aactgagctg acggagcacg agcacggaag ccatgcttgt 60 tgtatacata acataagccg gggggaggat atgctcgagg cattctcttc ttcctcatcc 120 gtcagtcact ggctcggtcc attcgttagc gtctcaccag tccttgatca gcattgttaa 180 240. tactactagc tcgctgctga gtgctgacta tgcgaaacag tttcttggca ggattccaac aactcgagct cgccgtcgcc gtcgctgtcg ctggaccgta cgaacgtgcc ggtcccctcc 300 ctgcatggcg gaccaaagag ccacccagat caggacggat ccccgtgaaa tcccccctct 360. tgttctttaa ttactcgcag gcggagga ggcggcagtg cacagcgaca gagagacga gactttgga tcgtccttgg gtgcatggat ggacggacgg acgaacgaga gggggggccg 480 cgagctctga atattcgccg ccgtcgatgc atcggcggcc tgcctgtcgc tctcgacgga 540 gagggtggta ctggtgtgcg caccggaca acgcaatgtt caggccaga gaatcggat cggaatatca tattccgtct ggtgtggtgg tcactttctt ttctttgtgt gtgtgttttt 660 ttgttgttgt tgtgtagata aatactatgg ggaagaatgg aggggatatg aggatatcct 720 cgttgattct gcttgagaaa ctagggcgta ttattgata catttggat tctcactctc ggctgggccg accccgccgg cgacctcagc ccgacgtgga father aagatgatta aagctttgta agataagact agtctgcact ttctagtaga tttagaccat attttcaaac 900 gtctaaaact aataatgaac agataaaact agctaagaga gaacgaatca gctaatagat 960 tagctaattg ttagttacat ttctcaaata gctattagtt gttagttaat ttaatctagc 1020 taaaatcaac tacaacaact tactcttt 1048 <210> 15 <211> 24 <212> DNA <213> Zea mays <400> 15 gatggatgga cggacgaacg aggc 24 <210> 16 <211> 30 <212> DNA <213> Zea mays <400> 16 catggatgga cggacggacg aacgagaggg 30 <210> 17 <211> 10105 <212> DNA <213> Zea mays <400> 17 cggtgtgtac aaagggcagg gacgtagtca acgcgagctg atgactcgcg cttactaggc 60 attcctcgtt gaagaccaac aattgcaatg atctatcccc atcacgatga aatttcccaa 120 gattacccgg gcctgtcggc caaggctata tactcgttgg atacatcagt gtagcgcgcg 180 tgccgcccag aacatctaag ggcatcacag acctgttat gcctcaaact tccgtggcct 240 aaacggccat agtccctcta agaagctaac tacggaggga tggctccgca tagctagtta 300 gcaggctgag gtctcgttcg ttaacggaat taaccagaca aatcgctcca ccaactaaga 360 acggccatgc accaccaccc atagaatcaa gaaagagctc tcagtctgtc aatccttgct 420 atgtctggac ctggtaagtt tccccgtgtt gagtcaaatt aagccgcagg ctccacgcct 480 ggtggtgccc ttccgtcaat tcctttaagt ttcagccttg cgaccatact ccccccggaa 540 cccaaagact ttgatttctc ataaggtgcc agcggggtcc tattagtaac acccgctgat 600 ccctggtcgg catcgtttat ggttgagact aggacggtat ctgatcgtct tcgagccccc 660 aactttcgtt cttgattaat gaaaacatcc ttggcaaatg ctttcgcagt tgttcgtctt 720 tcataaatcc aagaatttca cctctgacta tgaaatacga atgcccccga ctgtccctat 780 taatcattac tccgatcccg aaggccaaca caataggacc ggaatcctat gatgttatcc 840 catgctaatg tatccagagc gatggcttgc tttgagcact ctaatttctt caaagtaacg 900 gcgccggagg cacgacccgg ccagttaagg ccaggagcgc atcgccggca gaagggtcga 960 gccggtcggt tctcgccgtg aggcggaccg gccggcccgg cccaaggtcc aactacgagc 1020 tttttaactg caacaactta aatatacgct attggagctg gaattaccgc ggctgctggc 1080 accagacttg ccctccaatg gatcctcgtt aagggattta gattgtactc attccaatta 1140 ccagacacta acgcgcccgg tattgttatt tattgtcact acctccccgt gtcaggattg 1200 ggtaatttgc gcgcctgctg ccttccttgg atgtggtagc cgtttctcag gctccctctc 1260 cggaatcgaa ccctaattct ccgtcacccg tcaccaccat ggtaggcccc tatcctacca 1320 tcgaaagttg atagggcaga aatttgaatg atgcgtcgcc ggcacgaagg ccgtgcgatc 1380 cgtcaagtta tcatgaatca tcggatcggc gggcagagcc cgcgtcagcc ttttatctaa 1440 taaatgcgcc cctcccggaa gtcggggttt gttgcacgta ttagctctag aattactacg 1500 gttatccgag tagcacgtac catcaaacaa actataactg atttaatgag ccattcgcag 1560 tttcacagtt cgaattagtt catacttgca catgcatggc ttaatctttg agacaagcat 1620 atgactactg gcaggatcaa ccaggtagca cgtcctcgca gacgggccag cgccggcctc 1680 cgcgcggagg cgtcgtgccg ggctggcagt cgttcattcg ggcggaccga ttcttgggcg 1740 cgtgacgcca acgcgtctcc ggccttcagc gtgagccaca tccgagacca aaagcgccag 1800 cgaggtgtcc tcggtgccgc cggccatagg ccgacggcgg cacgaggcaa acgccgcgag 1860 cgctctcgag ccgacgagcc gcaccccggg gggtgagctc gacgaaggca acgtgtatcg 1920 agcacggctt cccgtgggac gggtagcagc acgcaagcac ttctcaacgc agcaggcata 1980 ggatgcccgc acgagcgatg ggacacaggc gccgggagtc ggccgcacgg cagcgggggt 2040 cctccaagca gtcacgggtc caagacaact catgcgcctg cgtagccgct acggtcgagc 2100 catccaaagc atccctccgc gctgggcgcg gcgggtctgc ttgcgaggac ggcgaccgaa 2160 ggtccaccga gcgcgggaga aacggaaaac gcatcgagca acgggccatc ccacggtgca 2220 gccactcgtc cagggcgtct ggccggcggt agccagccat agccggtcgt ggctgcgtca 2280 cggccgaacc acggccggcc aggcagccaa cagcgccagc cggagctggg cgcggtaggg 2340 tgccgaccgg ccacggctag gccgcgaggg ggtgcggggc tcggccgagg agacctggag 2400 gagacgctgg aaacgctatg gtttcagcag cgtttcgccc gggtttcggc tgcacgagtt 2460 ccctacccct actatacctg aggggcatac cccctcccag gacttcgggg agttctgcct 2520 tcagaaaacc agggcatttt cccagtaccc cacgaaaccc atctaagatg gctggacaca 2580 gcgtttttgc tcagaatcag gggtttcgct agcgtgaccc gttttccctc acgggtgcac 2640 ccgaacttcc acgtctcacg cggggcgacc acgggagggt cccgtgccct tccacgcgcc 2700 cgttttcgcg gccgtggccg aaaatccgtt tttggcccgt tcgccatggc gaacccctcg 2760 ttttcagcca aaacgcaagg ccgaacagcc ctgccgcccg ttgccttgcg tctcctcccg 2820 ttttccctcc gttccaccgt gcctttcaac cgagacctac gtagcaggct cggtgtcttt 2880 ccacgcgctt ggacttagcc cgttttcgcg gccgtggctg aaccgctgat ttcggccagc 2940 gcgccatggc gaacacctcg ttttcggccc agacgcaagg ccgaacagcc ctgccgcccg 3000 tcgccgcgcg cctcctcccg ttttccctcc gttccacctt gaccttcact ccagacatgc 3060 gctctaggtt cggaataaat atttgattt accatatctc gcttgggcaa tgtttcatat 3120 gttatgcatg tagtatggac atggctctca aacctacaga cctagcatct tttttaaa 3180 aaatagcgtc agcatacata ggcccttaa cccttgctg tgtaatccg tgagcatcct 3240 agctaggtgc ctaggtgagg aggtaccatg gttaatctat actactgat agcagaatc 3300 aaactctc aacaactact cccctccctcc gttctctatc ttatatcat aaggtttat 3360 cctaaatca tattttaaaa ctttaatagt tattaaaa tattcataa tattaacta 3420 aaaaaata actattaca tttatta aaacatctac ataacata aaaaaata 3480 ttttttat agagattgtt agtcaagtt ttaaaatttt gacttagaac aaccttcgt 3540 gaccttaaga tagggacag atgaagtatt atatatgctg ccatgcccca tcgaaacaaa 3600 tcaagtgata ccatcaat agctgcaaca gctgaaaact tgaaagggaa atgtgccttt 3660 gggccatttc taagtatttt tggtgattta gtgtctaaca caagtgccta agtgttgatc 3720 tatgcaagt gtggacaa gtgtaaatca agtcaaagg tatgtttcta gacttagtac 3780 attgttttat ggactgatgt attgtgtcta agtgctggaa acaggagaaa tcaaattgga 3840 aaagagatgt ctttgttcag ccaaagtctg ggtgcaccgg actgtccggt ggtgcaccag 3900 acagtgtccg gtgcgccagg cagactcagg cgaacttgct gctctcggga agtaattaac 3960 ggcgtacggc taaaattcac cggactgtcc ggtgagccaa cggtcggcca ggccaacagt 4020 cggccgggcc aacggtcggc cgcgcgatcc gcgcaggaca cgtggccgag ccaacggcta 4080 gtaggggcac cggactgtcc ggtgtggacc agacagtgtc cgatgcgcca acggctccaa 4140 ggctgccaac ggtcggcttc gccaaataag gaaggaaatc cgcaccggac tgtgcggtgg 4200 tgcaccggac agtccggtgc gccaggcgac agaaggcaag aattgccttc ccagattgct 4260 ctcaacggct cctagctgcc ttggggctat aaaagggacc cctaggcgca tggaggaaag 4320 aaccaagcat cctttgagca ttgttgatca ctcacactcc gttcttgcgc acttgttcga 4380 cattcttagt gatttgagct ccgttctagt gtgaaacttg tgatagtctc ttgagctcaa 4440 gtctgggtct tgtgtgtgcg tatttgctgt gatctttgtg tcttgtgtga gttgctcatc 4500 cctcccttac ttcgtgcttc tttgtgaaca tcaaagtgta agggcgagag gctccaagtt 4560 gtggagattc ctcgcgaacg ggatatagaa aagaaaagca aaacaccatg gtattcaagt 4620 gggtctttgg accgcttgag aggggttgat tgcaaccctc gtccgttggg acgccacaac 4680 gtggaagtag gcaagtgttg tacttggccg aaccacagga taaaccactg tgtctatttg 4740 tgttgattct gttgtggtta ttgtgtttcg ctaagactct tctctagcca cttggcatta 4800 ctgtgctaac gctgagagca cctagagggg ggggtgaata ggtgatcctg taaaacttaa 4860 acttatagcc acagaaactt ggttaatcgt tagcacaata attgccaagt ggctagagag 4920 gagtcaaaac acaataacca caagaaatca atcacagaga tgacacggtg gttatcccgt 4980 ggttcggcca agtacaaaac ttgcctactc cacgttgtgg cgtcccaacg gacgagagtt 5040 gcactcaact cctctcaagt gatccaatga tcaacttgaa taccacggtg ttcttcttta 5100 ctttgatctt ttcccgtttg cgaggaatct ccacaacttg gagtctctcg cccttacaat 5160 tgaatttcac aaagaagcac ggagtaaggg agggaagcaa cacacacaaa tccacagcaa 5220 tatgcgcaca cacacggcca agaatcgagc tcaaaagact atctcaaaat tctcactaga 5280 acggagctcg aattactgag aatgacaaat gaatgcgcaa agactgagtg tggatgatca 5340 agaatgctct aaggttgctt ggataactcc tccatgcgcc taggggtccc ttttatagcc 5400 ccaaggcagc taggagccgt tgagagcaaa tctggaagac caatcttgcc ttctgtcgtc 5460 gggtgcaccg gacaatccgg tgcacaccgg acactgtccg gtgcccgatt tctttcctta 5520 aacggcgcag tcgaccgttg ccgaccgttg cagatctggg agccgttggc gcaccggaca 5580 tgtccggtgc acaccggaca gtccggtgcc cccttccgac cgttggccag gccacgtgtc 5640 gcgcgcagat tccgcggccg accgttggct cggccgaccg ttggctcacc ggacagtccg 5700 gtgcacaccg gacagtccgg tgaattatag ccgtacgcca tcggcgaatt cccgagagcg 5760 gccacttcgc gccgtgtcag cctggcgcac cggacactgt ccggtgcacc accggacagt 5820 ccggtgtgct agaccgagct gagtcttggc tgtacacagc caagtctttg cacctttctt 5880 cttttctttt tctttctgtt tctaacactt agacaagtat attagtacac aaaaccaatg 5940 tactaaggct tagaacata cctttactca tgatttgcac tttgttcatc catgggcata 6000 gattcacatt taagcacttg tgttggcact caatcaccaa aatactttag aaatggccca 6060 aaggcacatt tccctttcaa tctccccctt tttggtgatt tatgccaaca caataaag 6120 6180 ggcatatatg gatcatcctt tgccaccact tggtttgttt ttgcaaatca aacacaaaat 6240 cctatctcta agtcaaatcc acttgtagag acaaaagag aggttttcca aagaaaattg 6300 attcaagatt ccaaaaactc cccctttttc ccataatcaa cacttctccc acaagagacc 6360 aacttttgac aaaagagaca atgcaagagt tttgaccaca caaaagctct aatctactat 6420 tttcaaaatt ctcaagtggt agctgatcca tttattgctt tggcctttat tttctccccc 6480 tttggcatca agcaccaaaa cgggattaat cttggcccta gaaccccatt gcctcaccaa 6540 aatcttcaac gaacaaa tagcaataag agttcatgag gtgaacttgg aataagttac 6600 cctctcatcg gagtgcagtg gaagtctttc atggtccaag tccacctttt ccctttcaat 6660 tctccttcga gactaaataa cgcaaactca agcatatggt tagtctcaaa agggtcaagt 6720 tgtaacacaa ctcccccaaa atatgtgcat cacttacaca aggacttgtg aggtccaggg 6780 aatgtttgta caacttgagc accacaataa gcaacaaaaa tgcagaatga acatgatcaa 6840 aggcataaac acatgtatgc tacaattcaa tccaagttcc gcgaatctaa gacatttagc 6900 tcactacgca gcctgcaaaa ggtcttctca tctagaggct tggtaaagat atcggctagc 6960 tggttctcgg tgctaacatg aaacacttcg atatctccct tttgctggtg gtctctcaaa 7020 aagtgatgcc ggatgtctat gtgctttgtg cggctgtgct caacaggatt ctccgccatg 7080 cggatagcac tctcattatc acataggagt gggactttgc tcagattgta gccaaagtcc 7140 cggagggttt gcctcatcca aagtagttgc gcgcaacact gtcctgcggc aacatactcg 7200 gcctcagcgg tggatagggc aacggaggtt tgtttcttag atttccacga caccagggac 7260 cttcctaaga attggcacgt ccctggtgta ctctttctat cgaccttaca tccagcatag 7320 tcggaatctg aatatccaat caagtcaaag gtagacccct ttggatacta gagcccgaag 7380 caaggcgtag caccaata tctaagaatt cgcttcaccg cactaagtg acactcctta ggatcggatt gaaatctagc acacatgcat acgctagca fatherccgg tctactagca cataaataaa gtaaagaccc tatcattgac cggtatgctt tttgatcaac ggacttacct cctttgttga ggtcggtgtg tccgtcggtc cccatcggag tctttgcggg cttggcgtcc 7620 ttcatcccaa accgctttag cagatcttgc gtgtacttcg tttggggagat gaaggtgccg tccttgagtt gcttcacttg gaacccaagg aagtagttca actcgcccat cattgacatc tcgaatttct gcgtcatcac cctgctaaac tcttcacaag acttttggtt agtagaacca aatattatgt catcgacata aatttggcac acaaacaaat caccatcaca tgtctttgta aaaagagttg gatcggcttt cccaaccttg aaagcattaa caattagaaa gtctctaagg cattcatacc atgctcttgg ggcttgctta agtccataga gcgccttaga gagcttacac 7980 acgtggtcgg ggtaccgttc atcctcgaag ccaggggggtt gctccacgta cacctcctcc 8040 ttgattggcc cattgagga agcgctcttc acatccattt ggaacaacct 8100. ttgattggcc tgagcggcat atgctagcaa buy gactctagcc tagccacagg agcaaaagtc tcctcaaagt ccaaacctgc gacttgggca taaccttttg ccacaagtcg agccttattc cttgtcacca ctccgtgctc gtcttgtttg ttgcggaca cccacttggt tcccacacaca ttttgcttag gacgaggcac cagcgtccaa acttcattgc gcttgaagtt gttgagttcc tcctgcatgg ccaacaccca gtccggatct agcaaggcct cttctatcct gaaaggctca 8460. 8460. 8460. 8460. 8460. 8460. 8460. 8460. 8460. 8460. 8460 cccttgctaa tgtcacccag aatttggttg acgggatgat ccctttgat catcgctcga acttgggttg gaggtgccgg ttccgcttct tcctccatca catgatcatc ttgtgctccc 8580 ccttgatcac acgcctcctg ttgatgaacc tgttcatcgt cttgagttgg gggatgcacc 8640 attgttgagg aagaaggttg atctcgttca tcttgttcct gtggccgaac ttctccaatc 8700. gccatggttc gaatagcggc cgtcggaaca tcttcttcat ctacatcatc aagatcaaca atttgctctc ttggagagcc attagtctca tcaaatacaa cgtcgctaga gacttcaacc aaacccgatg atttgttgaa gactctatac gcctttgtat ttgagtcata acctaacaaa 8880 aacccttcta cagctttggg agcaaactta gaatttctac ccttctttac tagaatgtag 8940 cacttgctcc caaatacacg aaagtaagat acattgggtt tgttaccggt tagtagctca 9000 tacgacgtct tcttgaggag gcgatgaagg tagaccctgt tgatggcgtg gcaagccgtg 9060 ttcacggctt cagtccaaaa gcactcgggg gtcttgaact ctcctagcat cgtcctcgcc 9120 atatcgatga gcgtcctgtt cttcctttct accacaccgt tttgctgtgg tgtgtaggga 9180 gcggagaact tgtgcttgat cccttcctct tcaaggaact cctccacttg aaggttcttg 9240 aactcggacc cgttgtcgct ccttatcttc tttactttga gctcaaactc attttgagct 9300 ctcctgagga agcgcttgag ggtcccttgt gtttcagact tatcctgcaa aaagaacacc 9360 caagtgaagc gggaaaagtc atcaacaata actagacctt acttacttcc tcctatgctc 9420 agataggcga cgggtccgaa gaggtccata tgtagcagct ccagtggtct tgaggtggtc 9480 atcacattct tgctgtgatg tgttcctccc acttgtttac ctgcttgaca cgctgcacaa 9540 ggtctatctt tttcgaattg aacgttagtc aaacctatta cgtgttctcc ctttagaagc 9600 ttgtgaaggt tcttcatccc cacatgtgct aagcggcgat gccacagcca gcccatgcta 9660 gtcttagcta ttaagcatgc atctagaccg gcctcttctt ttgcaaaatc aactaaataa 9720 agtttgtcgt ctaatacacc cttaaaagct actgaaccat cacttcttct aaagacagac 9780 acatctacat ttgtaaatag acagttatat cccatattgc ataattgact aaccgatagc 9840 aaattatatc ctagagactc tactaaaaac acattagaga tagagtgctc attagagatt 9900 gcaattttac ctaacccttt tatcttgcct tgattcccat caccgaatat gattgaatct 9960 tgggaatcct tattcttgac gtaggaggtg aacatcttct tctcccccgt catatggttt 10020 gtgcatccgc tgtcaataat ccagcttgaa cccccggatg cataaacctg caaggcaaat 10080 ttaggcttgg gttttaggta cccaa 10105 <21 O> 18 <211> 1652 <212> DNA <213> Zea mays <400> 18 cggtgtgtac aaagggcagg gacgtagtca acgcgagctg atgactcgcg cttactaggc 60 attcctcgtt gaagaccaac aattgcaatg atctatcccc atcacgatga aatttcccaa 120 gattacccgg gcctgtcggc caaggctata tactcgttgg atacatcagt gtagcgcgcg 180 tgccgcccag aacatctaag ggcatcacag acctgttatt gcctcaaact tccgtggcct 240 aaacggccat agtccctcta agaagctaac tacggaggga tggctccgca tagctagtta 300 gcaggctgag gtctcgttcg ttaacggaat taaccagaca aatcgctcca ccaactaaga 360 acggccatgc accaccaccc atagaatcaa gaaagagctc tcagtctgtc aatccttgct 420 atgtctggac ctggtaagtt tccccgtgtt gagtcaaatt aagccgcagg ctccacgcct 480 ggtggtgccc ttccgtcaat tcctttaagt ttcagccttg cgaccatact ccccccggaa 540 cccaaagact ttgatttctc ataaggtgcc agcggggtcc tattagtaac acccgctgat 600 ccctggtcgg catcgttat ggttgagact aggacggtat ctgatcgtct tcgagccccc 660 aactttcgtt cttgattaat gaaaacatcc ttggcaaatg cttcgcagt tgttcgtctt 720 tcataaatcc aagaatttca cctctgacta tgaaatacga atgcccccga ctgtccctat 780 taatcattac tccgatcccg aaggccaaca caataggacc ggaatcctat gatgttatcc 840 catgctaatg tatccagagc gatggcttgc tttgagcact ctaatttctt caaagtaacg 900 gcgccggagg cacgacccgg ccagttaagg ccaggagcgc atcgccggca gaagggtcga 960 gccggtcggt tctcgccgtg aggcggaccg gccggcccgg cccaaggtcc aactacgagc 1020 tttttaactg caacaactta aatatacgct attggagctg gaattaccgc ggctgctggc 1080 accagacttg ccctccaatg gatcctcgtt aagggattta gattgtactc attccaatta 1140 ccagacacta acgcgcccgg tattgttatt tattgtcact acctccccgt gtcaggattg 1200 ggtaatttgc gcgcctgctg ccttccttgg atgtggtagc cgtttctcag gctccctctc 1260 cggaatcgaa ccctaattct ccgtcacccg tcaccaccat ggtaggcccc tatcctacca 1320 tcgaaagttg atagggcaga aatttgaatg atgcgtcgcc ggcacgaagg ccgtgcgatc 1380 cgtcaagtta tcatgaatca tcggatcggc gggcagagcc cgcgtcagcc ttttatctaa 1440 taaatgcgcc cctcccggaa gtcggggttt gttgcacgta ttagctctag aattactacg 1500 gttatccgag tagcacgtac catcaaacaa actataactg atttaatgag ccattcgcag 1560 tttcacagtt cgaattagtt catacttgca catgcatggc ttaatctttg agacaagcat 1620 atgactactg gcaggatcaa ccaggtaccc aa 1652 <210> 19 <211> 10674 <212> DNA <213> Zea mays <220> <221> Unclassified feature <222> (3089)..(3188) <223> n is a, c, g, or t <400> 19 ccgacgccgc cggagatttt atctcgccgc cgttccacac acaccgcgac gtggacagcc 60 agcaccgctg ttattcttga accacggtat gagttcgttt gcttgcaatt gcaatagtcc 120 agcttctagt ttgttcgatc tatgtgcgta ttggcctgtg gtagtttctt tcttaggccg 180 tcgtttgggc ggggcgcgcc aggaaggagc agtctgcatc taataatcac ttgaacccaa 240 ttcaattcag tatatacaat tcttcttata tagacggaga catccatcta ctttcatcta 300 acatcatttt acacctgttt tgcttaaatt ctagatatac ttattacact tagcttgtga 360 ttcaacaggt accgtaccgt catgggcatg ttgatgaaca acgacgacag cagcagcagc 420 aggactagca tgcatccacg gccgcaggtc cttgcttccc tgcccttgct ggtctacgaa 480 tacgaggatc atcccaatgc cacaacaagg atgctcatat atagcctacc cgagcggagc 540 attgtctaca cgcacaacag cagtaggccc cagatgatga tgatggaggg taacttatcc 600 ttcagcaccc ctcaaggatg gctggtcatc cttggacaag cgtcagaggc ctcgatctgg 660 catccgctca ccggagagac catcacgctc ccaccaatac acggcgacca ccgtatcccc 720 gatagctgca agtgtctgct cacccgcagc tccgtcgccc acccggactg cgccgtcgtg 780 cttctcgacg tcaacgatcc tctcatgtgg ttctgccggg tgaatggcgg ggccgacagg 840 atgtgggtgc agcacgccta cgacattggc gaccactact tccccgagga gttccgcact 900 ccctccacgc ccaccaagaa tgttgtcgac gacgtcgccg cgctgggagg gaagctgtat 960 tttcgcttca ccgaatcaga ccaagacttc atgggcgtct ttgacttcga tttccatggc 1020 catactcccg ccgtggagtt ttacgagttt gatgtctccg aagagttcaa cctcaagttt 1080 cccgagggcg tgtgctctgc ctccatccac ttggtggagt ccatggatga gctttttgct 1140 gtctgcatct tctatgcga ttttgatccc accaacatta gcgccgctca tatcttcaag 1200 atggaggaaa tctgcgacga ggaacccgtg gcctggcacc gggtggatga tattggcgac 1260 agggctttcc tcctgacggg caccaacatg tcaacttggt gctctgcaag cacgaataac 1320 ctgaaaggga actccctcta ctttctaggc cacttagtag ctggccacag gaatctctgc 1380 atctatgata ttcaggagca atccatggag attgtccagg ttcacgacca agaagatatg 1440 gagatcgtgc gcacaccgcc atactggatt aatgtacctc cgtgctagta ttagcctatt 1500 acaatgtaat ttgcttaatt aatgtagctt gcttgctagc tagtgctatt tactgctgcc 1560 actactatat agtatattga atcaataaaa atagagtttg ctcgggtttt tgcattatcc 1620 gtgtatgaag ttttagaaga ggatttagca tctcaattttt ttttgggga caatgtacat 1680 catctatagg tatgtgtttg cactgaataa ggggtgattg gttctgtagc acaggtcttc 1740 ttccattgtg gcctaaaaga ctcagcaata ccagccctgt cctataagc atgtcatatt 1800 caaggagata atacttttt tattactctc ttcctttttt cggtggtaaa aaaacccctc 1860 ccatctcttc tttaataatt agagggggct acaatttctc gtttcgttaa tgtctactgt 1920 agtgcacagt ctctgtgatt tggtacagta ctgttttctg aaatctcatg gtttgttcat 1980 atcagtggtg atgcggtacc tctgtgttat gtgctagcgc ttgtttcatt agctagcctt 2040 agcttaaggc gtccagtaaa tattatattg tctgtgaatt ttatgtgctt caggttggtg 2100 tattattggt gctagattct gaagaacttg ccacaaaatg gagattgtcc aggttcacga 2160 ccagaaagat ttggaaatcg tgcgcacaca gccatcctgg attgcgata ccgccatgct 2220 aagtcgatta caatgtactt gcttaattaa tgtaggttgc tactagtgat tgcgctaatt 2280 cggccggctg cttctctcat ggtcgtccgc tcacttggga actgacgttt acaagacaga 2340 actacttcta ctagggataa caagcacggg tgaactgttg ttgaggccgg cttgcagtca 2400 tcctgtagcg aagcgtcaat cacttgcagt agtatctcgt ctgggaagct cctctcgacc 2460 aagctagtaa tgctgagtac gttttcaatc atagggtccg ttggtctctt cccggtcaac 2520 atctgtaaaa gtaaaactcc aaaactgtac acatccaagt gaggttgtgc tgtatgctgc 2580 <h2 style=";text-align:left;direction:ltr">gtctcgtcca gtcgtcgttg tggtacgata gaagctcgca atgccgaagt cccctagacg 2640<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> agcgttcatg tcatatcatc gaggaggaca ttactcggct tcacatcacc ttcaaatcta 2700<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ttccccgcta gggtcagaga tgacaaccga gtaagatcca atggcggacc taggtttttt 2760<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tttttttgta tagggtatgc ctcaataaaa cttttacata caattctata taatatactc 2820<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> catcggttcc aaaatagtat tacttttagc tcttggcttt tatgtcaaca ttcaaatgta 2880<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tagcgatgaa tctagacaca taaaatacat acaacaaaca ttttatgaac caattaatta 2940<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> cctaaaacga attttaattt aggatagaga gagagtacat ataattttat agtaaattca 3000<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ataaccaact cggtaaatag aaataaagtc atagtacatc aactagtaac aagacaacaa 3060<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ttaacaacgc catctaagtc gctgcatann nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 3120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 3180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> nnnnnnnnat gtcttggtga agggcacggt ggaacggagg gaaaacgaca cggaggcacg 3240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> cgacgacggg cggcaggggc gttcggcctt gcgtctgggc tggaaacgag gggttcgcca 3300<h2 style=";text-align:left;direction:ltr"> tggcgcgcgg gccgaaaacg gaggcttggg cacgaactcg aaaataagct aagtccaagc gtgtggaag acaccgaacc taaagtgcat gtcttgagtg aagggcaagg tggaacggag ggaaaattgg aggaggcgcg cctcgacggg cggcagggcc gttcggcctt gcgtattggc 3480 tgaaaacgag gggttcgcca tggcgcgcgg gccgaaaaaa acggttcggc cacggccgcg 3540 aaaacgagct aagtcccggc gtgtggaag acaccgaacc tagcgcat gtctggagtg aaggtgaagg tggaacggag ggaaaacggg aggaggcgcg cggcgacggg cggcagggct gttcggcctt gcgtatgggc tgaaaacgag gtgttcgcca tggcgcgcgg gccgaaaaca 3720 acggttcggc cacggccgcg aaaacgggct aagtccaagc gcgtggaag acaccgaggc 3780 tgctacgtag gtctcggttg aagggcacgg tggaacggag gaaaaacggg aggagacgca 3840 aggcaacagg cggcagggct gttcggcctt gcgttttggg tgaaaacgag gggttcgcca tggcgaacgg gccaaaaacg gatttttggc cacggccgcg aaaacgggca cgtggaaggg 3960. cacaggaccc tcccgtggtc accccgcgtg agacgtgga gttcgggtcc acccgtgagg 4020 gaaaacgggt cacgctagcg aaacccctga ttctgagcaa aaacgctgtg tccagccatc 4080 ttagatgggt ttcgtggggt actgggaaaa tgccctggtt ttctgaaggc agaactcccc 4140 gaagtcctgg gagggggtat gcccctcagg tatagtaggg ggtagggaac tcgtgcagcc 4200 gaaacccggg cgaaacgctg ctgaaaccat agcgtttcca gcgtctcctc caggtctcct 4260 cggccgagcc ccgcaccccc tcgcggccta gccgtggccg gtcggcaccc taccgcgccc 4320 agctccggct ggcgctgttg gctgcctggc cggccgtggt tcggccgtga cgcagccacg 4380 accggctatg gctggctacc gccggccaga cgccctggac gagtggctgc accgtgggat 4440 ggcccgttgc tcgatgcgtt ttccgtttct cccgcgctcg gtggaccttc ggtcgccgtc 4500 ctcgcaagca gacccgccgc gcccagcgcg gagggatgct ttggatggcc cgaccgtagc 4560 ggctacgctg gcgcatgagt tgtcttggac ccgtgactgc ttggaggacc cccgctgccg 4620 tgcggccgac tcccggcgcc cgtgtcccat cgctcgtgcg ggcatcctgt gcctgctgcg 4680 ttgagaagtg cttgcgtgct gctacccgtc ccacgggaag ccgtgctcga tacacgttgc 4740 cttcgtcgag ctcacccccc ggggtgcggc tcgtcggctc gagacgccc gcggcgtttg 4800 cctcgtgccg ccgtcggcct atggccggcg gcaccgagga cacctcgctg gcgctttttg 4860 tctcggatgt ggctcacgct gaaggccgga gacgcgttgg cgtcacgcgc ccaagaatcg 4920 gtccgcccga atgaacgacg gccagcccgg cacgacgcct ccgcgcggag gcggcgctg 4980 gccgtctgc gaggacgtgc tacctggttg atcctgccag tagtcatatg cttgtctcaa 5040 agattaagcc atgcatgtgc aagtatgaac taattcgaac tgtgaaactg cgaatggctc 5100 attaaatcag ttatagtttg tttgatggta cgtgctactc ggataaccgt agtaattcta 5160 gagctaatac gtgcaacaaa ccccgacttc cgggaggggc gcatttatta gataaaaggc 5220 tgacgcgggc tctgcccgcc gatccgatga ttcatgataa cttgacggat cgcacggcct 5280 tcgtgccggc gacgcatcat tcaaatttct gccctatcaa ctttcgatgg taggataggg 5340 gcctaccatg gtggtgacgg gtgacggaga attagggttc gattccggag agggagcctg 5400 agaaacggct accacatcca aggaaggcag caggcgcgca aattacccaa tcctgacacg 5460 gggaggtagt gacaataaat aacaataccg ggcgcgttag tgtctggtaa ttggaatgag 5520 tacaatctaa atcccttaac gaggatccat tggagggcaa gtctggtgcc agcagccgcg 5580 gtaattccag ctccaatagc gtatatttaa gttgttgcag ttaaaaagct cgtagttgga 5640 ccttgggccg ggccggccgg tccgcctcac ggcgagaacc gaccggctcg acccttctgc 5700 cggcgatgcg ctcctggcct taactggccg ggtcgtgcct ccggcgccgt tactttgaag 5760 aaattagagt gctcaaagca agccatcgct ctggatacat tagcatggga taacatcata 5820 ggattccggt cctattgtgt tggccttcgg gatcggagta atgattaata gggacagtcg 5880 ggggcattcg tatttcatag tcagaggtga aattcttgga tttatgaaag acgaacaact 5940 gcgaaagcat ttgccaagga tgttttcatt aatcaagaac gaaagttggg ggctcgaaga 6000 cgatcagata ccgtcctagt ctcaaccata aacgatgccg accagggatc agcgggtgtt 6060 actaatagga ccccgctggc accttatgag aaatcaaagt ctttgggttc cggggggagt 6120 atggtcgcaa ggctgaaact taaaggaatt gacggaaggg caccaccagg cgtggagcct 6180 gcggcttaat ttgactcaac acggggaaac ttaccaggtc cagacatagc aaggattgac 6240 agactgagag ctctttcttg attctatggg tggtggtgca tggccgttct tagttggtgg 6300 agcgatttgt ctggttaatt ccgttaacga acgagacctc agcctgctaa ctagctatgc 6360 ggagccatcc ctccatagtt agcttcttag agggactatg gccgtttagg ccacggaagt 6420 ttgaggcaat aacaggtctg tgatgccctt agatgttctg ggccgcacgc gcgctacact 6480 gatgtatcca acgagtatat agccttggcc gacaggcccg ggtaatcttg ggaaatttca 6540 tcgtgatggg gatagatcat tgcaattgtt ggtcttcaac gaggaatgcc tagtaagcgc 6600 gagtcatcag ctccgttgac tacgtccctg ccctttgtac acaccgcccg tcgctcctac 6660 cgattgaatg gtccggtgaa gtgttcggat cacggcgacg ggggcggttc gccgcccccg 6720 acgtcgcgag aagtccattg aaccttatca tttagaggaa ggagaagtcg taacaaggtt 6780 tccgtaggtg aacctgcgga aggatcattg ccgtgaccct taaacaaaac agaccgcgaa 6840 cgagtcaccc gtgccgccgg gctccggccc ggcacgctgc cccccccgaa cctcccgcgg 6900 ggaagggggg tgccgcgaaa aagaacccac ggcgccccgg gcgccaagga acaccagtac tacctcctgc cccgcggagc ggtcggcccg ccttccgctc ccagggcagc ggttacacct 7020 father gactctcggc aacggatatc tcggctctcg catcgatga gacgtagca aaatgcgata cctggtgtga attgcagaat cccgcgaacc atcgagtttt tgaacgcaag ttgcgcccga agccttctgg cggagggcac gtctgcctgg gcgtcacgcc aaaagacact 7200 cccaacaccc ccccgcgggg cgagggacgt ggcgtctggc cccccgcgcc gcagggcgag 7260 gtgggccgaa gcaggggctg ccggcgaacc gcgccgggcg cagcacgtgg tgggcgacat 7320 caagttgttc tcggtgcagc gtcacggcgc gcggccggac attcggccct aaggacccat 7380 cgagcgaccg agcttgccct cggaccgcga ccccaggtca gtcgggacta cccgctgagt 7440 ttaagcatat aataagcgg aggagaaga acttacgagg attcccctag taacggcgag cgaaccggga gcagcccagc ttgagaatcg ggcggcctcg ccgccgat tgtagtctgg 7560. agaggcgtcc tcagcgacgg accgggccca agttctctgg aaagggacgc ctgggagggt 7620. gagagccccg tccggcccgg accctgtcgc accacgaggc gccgtcaacg agtcgggttg 7680 tttgggaatg cagcccaaat cgggcggtaa actccgtcca aggctaaata caggcgagag 7740 accgatagcg aacaagtacc gcgagggaaa gatgaaaagg actttgaaaa gagagtcaaa 7800 gagtgcttga aattgccggg agggaagcgg atgggggctg gcgacgcgca ccggccgtat 7860 gcggaacggc tcctgctggt ccgccgatcg gctcggggcg tggaccgttg tcgcccgcgc 7920 cggcggccaa agcccggggg ccctaggcgc ccccggcagc cgtcgtcggc gcggacggta 7980 tccgcgcgcc tctggcgcgc ccctcggggc gctgcgccgc aacggcctgc gagctcccca 8040 tccgacccgt cttgaaacac ggaccaagga gtctgacatg cgtgcgagtc gacgggttca 8100 gaaacctgag atgcgcaagg aagctgacga gcgggaggcc ctcacgggcc gcaccgctgg 8160 ccgaccctga tcttctgtga agggttcgag ttggagcacg cctgtcggga cccgaaagat 8220 ggtgaactat gcctgagcgg ggcgaagcca gaggaaactc tggtggaggc tcgaagcgat 8280 actgacgtgc aaatcgttcg tctgacttgg gtataggggc gaaagactaa tcgaaccatc 8340 tagtagctgg ttccctccga agttttccctc aggatagctg gagcccacac gagttctatc 8400 gggtaaagcc aatgattaga ggcatcgggg gcgcaacgcc ctcgacctat tctcaaactt taaataggta ggacggcgcg gctgcttcgg tgagccgtgc cacggaatcg ggagctccaa gtgggccatt tttggtaagc agaactggcg atgcgggatg aaccggaagc cgggttacgg 8580. tgccaaactg cgcgctaacc tagacccac aaagggtgtt ggtcgataa gacagcagga cggtggtcat ggagtcga atccgctaag ggagtgtgta caactcacct gccgaatcaa ctagccccga aaatggatgg cgctgaagcg cgcgacccac acccggccat ctgggcgagc 8760. gacatgcccc gatgagtagg agggcgcggc ggccgccgca aaacccgggg cgcgagcccg 8820 ggcggagcgg ccgtcggtgc agatcttggt ggtagtagca aatattcaaa tgagaacttt 8880. gaaggccgaa gaggagaaag gttccatgtg aacggcactt gcacatgggt aagccgatcc tagggacgg gggaaacccg gcagatagcg cgatcaccg cgtcacccga aagggaatcg ggttaagatt tcccgagccg ggacgtggcg gcagacggcg acgttagga gtccggagac 9060 gccggcgggg gcctcggggaa gagttacttt ttctgcttaa cggcccgcca acctggaat 9120 cggttcagcc ggaggtaggg tccagcggcc ggaagagcac cgcacatcgc gcggtgtccg 9180 gtgcgccccc ggcggccctt gaaaatccgg aggaccgaat accgtccacg cccggtcgta 9240 ctcataaccg catcaggtct ccaaggtgaa cagcctctgg ccaatggaac aatgtaggca 9300 agggaagtcg gcaaaacgga tccgtaactt cgggaaaagg attggctctg agggttgggc 9360 tcgggggtcc cggccccgaa cccgtcggct gctggcggaa tgctcgagct gctcgcgcgg 9420 cgagagcggg ccgccgcgtg ccggccgggg gacggaccgg gaacggcccc ctcgggggcc 9480 ttccccggc gtcgaacaac cgactcagaa ctggtacgga caaggggaat ccgactgttt 9540 aattaaaaca aagcattgcg atggtcctcg aggatgctga cgcaatgtga tttctgccca 9600 gtgctctgaa tgtcaaagtg aagaaattca accaagcgcg ggtaaacggc gggagtaact 9660 atgactctct taaggtagcc aaatgcctcg tcatctaatt agtgacgcgc atgaatggat 9720 taacgagatt cccactgtcc ctgtctacta tccagcgaaa ccacagccaa gggaacgggc 9780 ttggcggaat cagcggggaa agaagaccct gttgagcttg actctagtcc gactttgtga 9840 aatgacttga gaggtgtagg ataagtggga gcctccgggc gcaagtgaaa taccactact 9900 tttaacgtta ttttacttat tccgtgggtc ggaagcgggg caccgcccct ccttttggct 9960 ccaaggcccg gcctcgccgg gccgatccgg gcggaagaca ttgtcaggtg gggagtttgg 10020 ctggggcggc acatctgtta aaagataacg caggtgtcct aagatgagct caacgagaac 10080 agaaatctcg tgtggaacaa aagggtaaaa gctcgtttga ttctgatttc cagtacgaat 10140 acgaaccgtg aaagcgtggc ctatcgatcc tttagacctt cggagtttga agctagaggt 10200 gtcagaaaag ttaccacagg gataactggc ttgtggcagc caagcgttca tagcgacgtt 10260 gctttttgat ccttcgatgt cggctcttcc tatcattgtg aagcagaatt caccaagtgt 10320 tggattgttc acccaccaat agggaacgtg agctgggttt agaccgtcgt gagacaggtt 10380 agttttaccc tactgatgac cgcgccgga tagtaattca acctagtacg agaggaaccg 10440 ttgattcaca caattggtca tcgcgcttg ttgaaaagcc agtggcgcga agctaccgtg 10500 tgccggatta tgactgaacg cctctaagtc agaatccaag ctagcaaccg gcgcctctgc 10560 tcgccgcccg ccccgaccca cgttagggcg ttcgcgcccc aagggcccgt gccattggct 10620 cagcccgccc ggccgacgcg ccgcggcggg ccgcctcgaa gctcccttcc caac 10674 <210> 20 <211> 674 <212> DNA <213> Zea mays <400> 20 tacctggttg atcctgccag tagtcatatg cttgtctcaa agattaagcc atgcatgtgc 60 aagtatgaac taattcgaac tgtgaaactg cgaatggctc attaaatcag ttatagtttg 120 tttgatggta cgtgctactc ggataaccgt agtaattcta gagctaatac gtgcaacaaa 180 ccccgacttc cgggaggggc gcatttatta gataaaaggc tgacgcgggc tctgcccgcc 240 gatccgatga ttcatgataa cttgacggat cgcacggcct tcgtgccggc gacgcatcat 300<00,01349>tcaaatttct gccctatcaa ctttcgatgg taggataggg gcctaccatg gtggtgacgg 360 gtgacggaga attagggttc gattccggag agggagcctg agaaacggct accacatcca 420 aggaaggcag caggcgcgca aattacccaa tcctgacacg gggaggtagt gacaataaat 480 aacaataccg ggcgcgttag tgtctggtaa ttggaatgag tacaatctaa atcccttaac 540 gaggatccat tggagggcaa gtctggtgcc agcagccgcg gtaattccag ctccaatagc 600 gtatatttaa gttgttgcag ttaaaaagct cgtagttgga ccttgggccg ggccggccgg 660 tccgcctcac ggcg 674 <210> 21 <211> 721 <212> DNA <213> Zea mays <400> 21 tacctggttg atcctgccag tagtcatatg cttgtctcaa agattaagcc atgcatgtgc 60 aagtatgaac taattcgaac tgtgaaactg cgaatggctc attaaatcag ttatagtttg 120 tttgatggta cgtgctactc ggataaccgt agtaattcta gagctaatac gtgcaacaaa 180 ccccgacttc cgggaggggc gcatttatta gataaaaggc tgacgcgggc tctgcccgcc 240 gatccgatga ttcatgataa cttgacggat cgcacggcct tcgtgccggc gacgcatcat 300 tcaaatttct gccctatcaa ctttcgatgg taggataggg gcctaccatg gtggtgacgg 360 gtgacggaga attagggttc gattccggag agggagcctg agaaacggct accacatcca 420 aggaaggcag caggcgcgca aattacccaa tcctgacacg gggaggtagt gacaataaat 480 aacaataccg ggcgcgttag tgtctggtaa ttggaatgag tacaatctaa atcccttaac 540 gaggatccat tggagggcaa gtctggtgcc agcagccgcg gtaattccag ctccaatagc 600 gtatatttaa gttgttgcag ttaaaaagct cgtagctcga cccttctgcc ggcgatgcgc 660 tcctggcctt aactggccgg gtcgtgcctc cggcgccgtt actttgaaga aattagagtg 720 c 721 <210> 22 <211> 10611 <212> DNA <213> Zea mays <400> 22 accacataaa aacattcccc ctagagtagc tgttaatacg aataacagaa actctgttat 60 agccatttct gtacattcaa tgtactctac ggatagagga atacataaag ttgaacataa 120 taaaataaga aattgaaaga tttcgttgaa attgttcgtt tggaaatttc ccgaaaagct 180 aattataggt tcttctctcc atcggaacaa tagggccgtt atgcttatta ctaaacttgt 240 tgaagagatg aatagaacc aaggtctatc tttttgatca gaggttaaat cgatcatcag 300 aagaagaatt aggccaaaaa ttaggataca ttctgggaaa atgaaacttc catggaagag 360 aagcaaatga aacgctttca taaaaattct cgtagaatcg agaatgaagt tttcattctg 420 tacatgccag atcatgaatt agtaactgca gccaatctcc gaaaagtccc gattgtttcg 480 atttttggaa tgggatattt acggaatccc catgaatagg atcaaacctt attccatgct 540 atttccataa gattcctctt tcttattctt aagcaagccc ccgagagggc ttagttgatc 600 atgatttctg ttttctcttt ttttcctttt ttatttgttt cgaaaaagat atcgtccgat 660 tctccttcta ttgattcttt tccgatcgag atgtatggat ccatgtgtct acatacctag 720 attctgttca tggattaacg aaaatgtgca agagctctat ttgcctctgc cattctatga 780 gtcgcttcct tttgcgtat ggcaccccca ctccctttgg cagcatctac taattcggaa 840 cttaatttga aagccatatt tcgacccgga cgcttttggg atgcttctaa taaccaacga 900 atggcaagtg ctcttccttg tttagatcct atttcaatcg gaactttccg cgtcgatcct 960 tttttattac gtcttgtttt tactcctata ttgggagtta ctctacgtat tgcttgacgt 1020 aaaaccaata gtggatttgt ttctgtcttt tgttgaatct ttttcacggc tcgatagaga 1080 atttgataag ccaatgattt ttttccgtct ttcataatac ggttaaccac catgttaact 1140 aatcgattac gaaaaattgg atcggatttt gcggttcttt tttctgcagt acctcgacgt 1200 gacatgagcg tgaaagaggt tcaagaatcc gttttctttt tataagggct aaaaacgaat 1260 cacttatttt tttggctttt tggccccata ttgtagggtg gatctcgaaa gataggaaag 1320 atctccctcc aagccgtaca tacgactttc atcgaatacg gctttccaca gaattctata 1380 gggatctatg agatcgagta tggaattctg tttactcact ttaaattgag tatccgtttc 1440 cctccttttc ccgctaggac cggaaatcct gtattttcca tatccatacg atcgagtcct 1500 taggtttccg aaatagtgta atggaaaaag aagtgcttcg aatcattgct atttgactcg 1560 gacctgttct gaaaaagtcg aggtatttcg aattgtttgt tgacacggac aaagtaaggg 1620 aaaacctctg aaagaatttc catattgacc ttggacatat aagagttccg aatcgaatct 1680 ctttagaaag aagatctttt gtctcatggt agcctgctcc agtcccctta cgaaactttc 1740 gttattgggt tagccataca cttcacatgt ttctagcgat tcacatggca tcatcaaatg 1800 atacaagtct tggataagaa tctacaacgc actagaacgc ccttgttgac gattctttac 1860 tgcgacagca tctagggttc ctcgaataat gcgatatctc acaccgggta aatccttaac 1920 ccttcctcct cttactaata ctacagaatg ttcttgtaaa ttatggccaa taccaggtat 1980 ataagcagtg atttcaaatc cagaggttaa tcgtactctg gcaactttac gtaaggcaga 2040 gttgggtttt ttggggttga tagtggaaaa gtcgacagat aagtcatcct tactgtccct 2100 ctacagaacc gtacatgaga ttttcacctc atacggctcc tcgttcaatt cttcgaagg 2160 gatccttttc ctcgttcgag agtctccgcc cttcttccac tccgcccga agactaacta 2220 agaccaattg agtcacgttt tcatgttcta attgaacact ttccatttat gattaaagga 2280 gaagattgtt cttttaccaa acatatgcgg atcaaatcac gtcttataat aagaagaaat 2340 cttctcggt atcaatcccc ttgcccctca ttctttgaga atcagaagga tccttttcga 2400 gtttccattt cttcatttg aatctgggct cttctatctt cgacttattt ttttggcttt 2460 attctttatt tatttcattt cgatttttcc ctcttcctct atccctatcc tctaggtaca 2520 gcgtttgcat caatagagaa ctttttcctc tgtatgaatc gatattattc caatttcttc 2580 ccgaaacttc ccaagaaaaa tcccgaattg gatccaaaat tgacgggtta atgtgagctt 2640 atccatgcgg ttaggcactc ttcaaatagg aatccatttt ctaactggct ttcgtgcttt 2700 ggtgagtcgt ccgagatcct ttcgatgacc tatgttgtgt tgaagggata tctatatgat 2760 ccgatcgatt gcataagacc cgcggtagca atagaacggg gaaagtatac agaaaagaca 2820 gttctttcg atttcgatta tctatatattt agttcgtttc tatttctaga tatctatttc 2880 tatatatcta tatattagta ttaatatcta tatattagta ttagttatct atatattagt 2940 attagttagt agtactattc tattagttag cgatcccggc tctgtgagtt cttcttccg 3000 tgatgaactg tcggcaccag tcctacattt tttctctgtg gaccgaggag aaagggggct 3060 cagcaggaag aggattgtac catgagagaa gcacagaggt caacccgctt caaatatgga 3120 acatggattc tggcaatgca acggagttgg gtcctcatat cgatccgaat gaatcagtct 3180 ttctacagag gtcaatcttt gcctattagg caagaggata gcaagttcga aattctgtct 3240 cggtaggaca tggatttcta ttactatgaa attcataaat tagttaatgg gggggctacc 3300 attatcctttt ttcttgtatg tgttcctaag agaaggaatt tgtccatttc atgtttcgag 3360 gtctcaaaaa aagggcgtgg aaacagatag aaactcttga atggaaattg aaaagaaatg 3420 tagccccagt tccttcggaa atggtaagat ctttggcgca agaagaaggg gcgatccata 3480 tcatcttgac ttggttctgc ttcccctcttt ttttaagaa taccgagtcg ggttcttctc 3540 ctaccagtat cgaatagaac atgctgaaca agatcttctt catggaaacc cactcgattt 3600 agatcgggaa aatcgtacag attttatgaa accatgtgct atggctcgaa tccatagtca 3660 atcctatttt cgataggacc ggttgacaat tgaatccaat ttttcccatt atttgactgt 3720 ccataatagt gcggaaagaa agcccggagg aagagtggcc ttgcgtttct cgcccctttg 3780 ccttaggatt cgttaattct ctttctcgat gggacgggga agggatataa ctcagcggta 3840 gagtgtcacc tgacgtgt ggaagtcatc agttcgagcc tgattccc taaacctaat 3900 gtgagtttt tctattttga cttactcccc caccacgatc gaacgggaat ggataagagg 3960 cttgtgggat tgacgtgata gggtaggtt ggctatactg ctggtggcga actccaggct 4020 aaatctga agcgcatgga tacaagttat ccttggagg aaagacatt ccgaatccgc 4080 ttgtctacg aaggaag ctataagtaa tgcaactatg attctcatgg agagttcgat 4140 cctggctcag gatgaacgct ggcggcatgc ttacacatg caagtcgac gggaagtggt 4200 gtttccagtg gcgaacggggt gagtaacgcg tagaacctg cccttgggag gggacacaca 4260 actggaaacg gttgctaata cccgtaggc tgaggagca aaggagaat ccgcccaagg 4320 agggctcgc gtctgattag ctagttggtg agcaatagc ttaccaggc gatgatcagt 4380 agctggtccg agaggatgat cagccacact gggactgaga cacggcccag actcctacgg 4440 gaggcagcag tgggaattt tccgcaatgg gcgaagcct gacggagca tgccgcgtgg 4500 aggtggaagg cctacgggtc gtcaacttct tttctcggag aagaaacaat gacggtatct 4560 gaggaataag catcggctaa ctctgtgcca gcagccgcgg taagacagag gatgcaagcg 4620 ttatccggaa tgattgggcg taaagcgtct gtaggtggct tttcaagtcc gccgtcaaat 4680 cccagggctc aaccctggac aggcggtgga aactaccaag ctggagtacg gtaggggcag 4740 agggaatttc cggtggagcg gtgaaatgca ttgagatcgg aaagaacacc aacggcgaaa 4800 gcactctgct gggccgacac tgacactgag agacgaaagc taggggagca aatgggatta 4860 gagaccccag tagtcctagc cgtaaacgat ggatactagg tgctgtgcga ctcgacccgt 4920 gcagtgctgt agctaacgcg ttaagtatcc cgcctgggga gtacgttcgc aagaatgaaa 4980 ctcaaaggaa ttgacgggg cccgcacaag cggtggagca tgtggtttaa ttcgatgcaa 5040 agcgaagaac cttaccaggg cttgacatgc cgcgaatcct cttgaaagag aggggtgccc 5100 tcgggaacgc ggacacaggt ggtgcatggc tgtcgtcagc tcgtgccgta aggtgttggg 5160 ttaagtctcg caacgagcgc aaccctcgtg tttagttgcc actatgagtt tggaaccctg 5220 aacagaccgc cggtgttaag ccggaggaag gagaggatga ggccaagtca tcatgcccct 5280 tatgccctgg gcgacacacg tgctacaatg ggcgggacaa agggtcgcga tctcgcgagg 5340 gtgagctaac tccaaaaacc cgtcctcagt tcggattgca ggctgcaact cgcctgcatg 5400 aagcaggaat cgctagtaat cgccggtcag ccatacggcg gtgaatccgt tcccgggcct 5460 tgtacacacc gcccgtcaca ctataggagc tggccaggtt tgaagtcatt acccttaacc 5520 gtaaggaggg ggatgcctaa ggctaggctt gcgactggag tgaagtcgta acaaggtagc 5580 cgtactggaa ggtgcggctg gatcacctcc tttcaggga gagctaatgc ttatgcttat 5640 tgggtatttt ggtttgacac tgcttcacgc ccaaaaagaa ggcagctacg tctgagctaa 5700 acttggatat ggaagtcttc tttcgtttag ggtgaagtaa gaccaagctc atgagcttat 5760 tatcctaggt cggaacaaat tagttgatag tgataggatc cccttttttga cgtccccatg 5820 tccccccgtg tggcggcatg gggatgtcaa aaggaaaggg atggagtttt tctcgcttt 5880 ggcgtagcag gcctcccttt gggaggccg cgcgacgggc tattagctca gtggtagac 5940 gcgcccctga taattgcgtc gttgtgcctg ggctgtgagg gctctcagcc acatggatag 6000 ttcaatgtgc tcatcagcgc ctgacccgaa gatgtggatc atccaaggca cattagcatg 6060 gcgtactcct cctgtttgaa tcggagtttg aaaccaaaca aacttctcct caggaggata 6120 gatggggcga ttcaggtgag atcccatgta gatcgaactt tctattcact cgtgggatcc 6180 gggcggtccg ggggggggcc accgcggctc ctctcttctc gagaatccat acatccctta 6240 tcagtgtatg gagagctatc tctcgagcac aggttgaggt tcgtcctcaa tgggaaaatg 6300 gagcacctaa caacgcatct tcacagacca agaactacga gatcacccct ttcattctgg 6360 ggtgacggag ggatcgtacc attcgagcct ttttttcatg cttttcccgg cggtctggag 6420 aaagcagtaa tcaataggac ttccctaatc ctcccttcct gaaaggaaga acgtgaaatt 6480 ctttttcctt tctgcaggga ccaggagatt ggatctagcc ataagaggaa tgcttggtat 6540 aaataagcca cttcttggtc ttcgaccccc taagtcacta cgagcgcccc cgatcagtgc 6600 aatgggatgt ggctatttat ctatctcttg actcgaaatg ggagcagagc aggtttgaaa 6660 aaggatctta gagtgtctag ggttgggcca ggagggtctc ttaacccctt cttttttctg 6720 cccatcggag ttattccca aggacttgcc gtggtaaggg ggagaagggg gaagaagcac 6780 acttgaagag cgcagtacaa cggggagttg tatgctgcgt tcgggaagga tgaatcgctc 6840 ccgaaaagga gtctattgat tctctcccaa ttggttggat cgtaggggcg atgatttact 6900 tcacgggcga ggtctctggt tcaagtccag gatggcccag ctgcgccagg gaaaagaata 6960 gaagaagcat ctgactcttt catgcatact ccacttggct cgggggggat atagctcagt 7020 tggtagagct ccgctcttgc aattgggtcg ttgcgattac gggttggctg tctaattgtc 7080 caggcggtaa tgatagtatc ttgtacctga accggtggct cactttttct aagtaatggg 7140 gaagaggact gaaacatgcc actgaaagac tctactgaga caaaaagatg ggctgtcaaa 7200 aaggtagagg aggtaggat ggcagttggt cagatctagt atggatcgta catggacgat 7260 agttggagtc ggcggctctc ctaggcttcc ctcatctggg atccctgggg aagaggatca 7320 agttggccct tgcgaatagc ttgatgcact atctcccttc aaccctttga gcgaaatgtg 7380 gcaaaaggaa ggaaaatcca tggaccgacc ccattgtctc caccccgtag gaactacgag 7440 atcaccccaa ggacgccttc ggcgtccagg ggtcacggac cgaccataga tcctgttcaa tagtggac acattagccg tccgctctcc ggttgggcag tagggtcgg agaagggcaa 7560 tcactcgttc ttaaaaccag cattcttaag tcactcgttc agagtcgggc ggaaaaaggg gagagctccc cgttcctggt tctcctgtag ctggattccc cggaccaca agaatcctta gaatgggatt ccaactcagc accttttgtt ttgggatttt gagaagagtt gctctttgga gagcacagta cgatgaaagt tgtaagctgt gttcgggggg gagttattgc ctatcgttgt 7800. cctctatggt agaacccgtc ggggaggcct gagaggcggt ggtttaccct gtggcggatg 7860 tcagcggttc gagtccgctt atctccagcc cgtgaactta gcggatacta tgatagcacc 7920 gaaggttgcc aattcgtcag ttcgatctat gattcgcat tcatggacgt tgataagatc cttccattta gtagcacctt aggatggcat agccttaacg ttaatggcga ggttcaaaag aggaaaggct tgcggtggat acctaggcac ccagagacga ggaagggcgt agcaagcgac 8100 gaatgcttc ggggagttga aataagcat agatccggag attcccaaat aggtcaacct tttgaactgc ctgctgaatc catgagcagg caagacaa cctggcgaac tgaaacatct 8220 tagtagccag aggaaaagaa agcaaaagcg attcccgtag tagcggcgag cgaaatggga 8280 gcagcctaaa ccgtgaaaac ggggttgtgg gagagcaata caagcgttgt gctgctaggc 8340 gaagcggttg agtgccgcac cctagatggc taagtccag tagccgaaag catcactagc 8400 ttacgctctg accccgagtag catggggcac gtggaatccc gtgtgaatca gcaaggacca 8460 ccttgcaagg ctaaatactc ctgggtgacc gatagcgaag tagtaccgtg agggaaaggt 8520 gaaaaacc cccagtgggt agtgaaatag aacgtgaaac cgtgctgagc tcccaagcag 8580 tgggaggga aagtgatctc tgaccgcgtg cctgttgaag aatgagccgg cgactcatag 8640 gcagtggctt ggttaaggga atggaaccca ccggagccgt agcgaaagcg agtcttcata 8700 gggcgattgt cactgcttat ggacccgaac ctgggtgatc tatccatgac caggatgaag 8760 cttggatgaa actaagcaga ggtccgaacc gactgatgtt gaagaatcag cggatgagtt 8820 gtggtaggg gtgaaatgcc actcgaaccc agagctagct ggttctcccc gaaatgcgtt 8880 gaggcgcagc agttgactgg acatctaggg gtaaagcact gtttcggtgc gggctgcgcg 8940 agcggtacca aatcgaggca aactctgaat actagatatg acccaaaaat aacaggggtc 9000 aaggtcggcc agtgagacga tgggggataa gcttcatcgt cgagagggaa acagcccgga 9060 tcaccagcta aggcccctaa atgaccgctc agtgataaag gaggtgggg tgcaaagaca 9120 gccaggaggt ttgcctagaa gcagccaccc tttaaagagt gcgtaatagc tcactgatcg 9180 agcgcccttg cgctgaagat gaacggggct aagcgatctg ccgaagctgt gggatgtcaa 9240 aatgcatcgg taggggagcg ttccgcctta gaggggaagca aacgcgaaag cgggggtcga 9300 cgaagcgaa gcgagaatgt cggcttgagt aacgaaaaca ttggtgagaa tccaatgccc 9360 cgaaaaccca aggtttcctc cgcaaggttc gtccacggag ggtgagtcag ggcctaagat 9420 caggccgaaa ggcgtagtcg atggacaaca ggtcaatatt cctgtactac cccttgttgg 9480 tacgggaggga cggaggaggc taggttagcc gaagatggt tataggttta aggacacaag 9540 gtgaccctgc ttttcaggg taagaagggg tagagaaaat gcctcgagcc gaggtccgag 9600 taccaagcgc tgcagcgctg aagtatgagc cccgtggact agccattgct tctccacgag 9660 gctcatacca ggcgctacgg cgctgaagta tgtaacccat gccatactcc caggaaaagc 9720 tcgaacgacc ttcaacaaaa gggtacctgt acccgaaacc gacacaggtg ggtaggtaga 9780 gaatacctag gggcgcgaga caactctctc taaggaactc ggcaaaatag ccccgtaact 9840 tcgggagaag gggtgccccc tcgcaaagg gggtcgcagt gaccaggccc gggcgactgt 9900 ttaccaaaaa cacaggtctc cgcaaagtcg taagaccatg tatggggct gacgcctgcc 9960 cagtgccgga aggtcaagga agttggtgaa ctgatgacag ggaagccggc gaccgaagcc 10020 ccggtgaacg gcggccgtaa ctataacggt cctaaggtag cgaaattcct tgtcgggtaa 10080 gttccgaccc gcacgaaagg cgtaacgatc tgggcactgt ctcggagaga ggctcggtga 10140 aatagacatg tctgtgaaga tgcggactac ctgcacctgg acagaaagac cctatgaagc 10200 tttactgttc cctgggattg gctttgggct tttcctgcgc agcttaggtg gaaggcgaag 10260 aaggccccct tccgggggg cccgagccat cagtgagata ccactctgga agagctcgga 10320 ttctaacctt gtgtcagacc cgcgggccaa gggacagtct caggtagaca gtttctatgg 10380 ggcgtaggcc tcccaaaagg taacggaggc gtgcaaaggt ttcctcgggc cagacggaca 10440 ttggtcctcg agtgcaaagg cagaagggag cttgactgca agactcaccc gtcgagcaga 10500 gacgaaagtc ggccttagtg atccgacggt gccgagtgga agggccgtcg ctcaacggat 10560 aaaagttact ctagggataa caggctgatc ttccccaaga gtccacatcg a 10611 <210> 23 <211> 611 <212> DNA <213> Zea mays <400> 23 cccgcacaag cggtggagca tgtggtttaa ttcgatgcaa agcgaagaac cttaccaggg 60 cttgacatgc cgcgaatcct cttgaaagag aggggtgccc tcgggaacgc ggacacaggt 120 ggtgcatggc tgtcgtcagc tcgtgccgta aggtgttggg ttaagtctcg caacgagcgc 180 aaccctcgtg tttagttgcc actatgagtt tggaaccctg aacagaccgc cggtgttaag 240 ccggaggaag gagaggatga ggccaagtca tcatgcccct tatgccctgg gcgacacacg 300 tgctacaatg ggcgggacaa agggtcgcga tctcgcgagg gtgagctaac tccaaaaacc 360 cgtcctcagt tcggattgca ggctgcaact cgcctgcatg aagcaggaat cgctagtaat 420 cgccggtcag ccatacggcg gtgaatccgt tcccgggcct tgtacacacc gcccgtcaca 480 ctataggagc tggccaggtt tgaagtcatt acccttaacc gtaaggaggg ggatgcctaa 540 ggctaggctt gcgactggag tgaagtcgta acaaggtagc cgtactggaa ggtgcggctg 600 gatcacctcc t 611 <210> 24 <211> 22902 <212> DNA <213> Zea mays <400> 24 attcattatt ggcccaccat tgattacaag atttagcttt tatgaatcgc tattggtttg 60 atacgaataa tggcagtcgt ttcagtttgt taaggataca gatgtatcca caattcattt 120 agagttactt aatagcctat ttcttatact atatctctat cccgtgaaat tctcaagccc 180 aaagatggat gcatatgctg tgtttcattt tgctaaatga tatcaattaa atggtatatc 240 aattctataa attggatata acaataaata aatcagaaaa attcttttat tttagataga 300 agaaatgttt cttctatcta aaataaatga atgtaccctt ctatccaaat ccaatttgca 360 tcgataaaat aaatccaaat tccagattct agcagtagat gaataattgc aaatttttgt 420 gtgtacgaga ttagaataac ttaaaaataa ctgacataat ttttattt tcctgaccaa 480 aaaaatacat gaaaaagaaaaaaaattgttg atttatggtt aaagaaaaaaaaaaaaaaaaaaaaaaaaa'dd by aaatttgttg attttggtt aacaagaaaa caggggttct gttgaatttc aagtattcag ttcactaat aagatacgga 600 gacttgcttc acatttagaa ttacacaaa atattttc atcggaaaga ggtctacgaa 660 gacttttggg aaacgtcaa agtttgctgg cttattttggt aaagaaaat agagtacgtt 720 ataagaaattt addaagtcag ttggatattc gggagaagta atttaatcgt tctcattttt 780 ttcttatttt attagtagtc ttatagtagt attagatttt gtattttgat gagcctcgtt 840 ttgaggaattt catggataa tccattttca tggaataag atagaaca aggatatgag 900 tctatcgctt aaagaaaag atctcatgat agtcaatg ggcactcaac acccatcaat 960 gcatggtgtt cttcgactga ttgttactct cgatggtga gatgttattg attgtgaacc 1020 catattagggg tattacaca gaggaatgga aaaaatcgcg gaaacagaa gttattaca 1080 atacttgcct tatgtacac gatgggatta atgtttacag aagcaatac 1140 ggtaaatgca ccagaattct tagagaata tcattaccc aaagagcca actatattag 1200 agtaattatg ttagaattga gccgtatagc ttctcattg ttatggctg gaccttttat 1260 ggaggatctc ggggcacaga ctcctttt ctacattttt agagagagag agaattgata 1320 tatgatctt ttgaagctgc tacaggtatg cgaatgatgc attackt tcgcatcgga 1380 ggagtcgctg ccgatctccc ttatgtgg atggataat gtttagattt ctgtgattat 1440 tttttacaag gagttgttga atcagaa cttattacac agaatcccat tttttagaa 1500 cgagttgaag gagtcggttt tattagcgga gagaagctg taattgggg cttatcggga 1560 ccaatgttac gagcttctgg atacaatgg gatcttcgta aaattgatcc ttatgagtct 1620 tacaatcaat tcgattggaa agtccaatgg caaaagaag gagattcgtt agctcgctat 1680 1740 ttcacaggc tcggtgaat gagggaatcc aaaaatta ttcacaggc tgtagagaaa 1740 attcctagag gaccttatga gatttagaa gcccgacgct ttaagaagc aaagaatccc 1800 gatggaatg attttgaata tcgatttctt ggtaaaaac cttcgccca ttctgaatta 1860 tcaagcaag agctttatgt aagagtagaa gctccaaag gcgaattagg atttatctg 1920 gtaggagatg atagcttttt cccctggaga tggaaaattc gtccaccggg ttttattaat 1980 ttgcaaattc ttcctcatct agttaaaaaa atgaattgg ctgatatcat gataatta 2040 ggtagtatag attcattat gggggaagtt gatcgttgaa atgataatag atagggtaga 2100 ggtagaaact atcaatctt ttcgaaatc agaattattt aaagaaatct acgaacttat 2160 atggattcta cccattttg cccctcctact gggaatcaca atagaagtac tcgtattgt 2220 gtggttagaa agagaatat ccgcatcgat acaacacgt attggtcctg atatgctgg 2280 ccccctggga ctgcttcaag ctatagcaga tggaactaa ctactttaa agaggatat 2340 cctcccatcc cgaggagata ttccttatt tagcattggt cctctatag cagtcatatc 2400 cattttatta agttttttag ttatcccttt aggatatcgt ttgttttag ctgatcttag 2460 tattggtgtt tttgat tgcgatttca agtattgctc ctattggtct tctcatggca 2520 ggatatagct caataataa atattctttt tcaggcggtc tacgagcggc tgctcaatct attagttatg aaataccatt aactttttgt gtactagcaa tatctctacg tgtgattcgt 2700. tctttttcct ctaaataca ttgaatgctt atcttccttt gcttattctg fathercgcgtt ggtaagttaa actcgatagc fathergagtg aaacaaaaca gcttattaat ttgtagtaaa agtaaaaaat ctcatttcct aacgtacaaga aaaaagttca agtaaacata agcagtgtaa actcttaacc ccaaggttga gattgtttga ttagtcatca tatcttgaag cgggcaagaa taaagattc gcgatatgga attccattac tagaatattt tgagttatta ctataattta aacttataac cacaaggcaa tcgatcaaaa tttagtgagg gattaggac actaaagtac attack attack act attack gaatctaaat cattagacat ttttcgtcat aaaaggaatc father cttgaaattg gtggaaatga tcaagccgta ctttcttcag attccggtct agagtatgtt cccattcact tgttaagga atggctatca agaacgaatt aaccctttat tcttttttta agtatacccc tcctagggaa agaagagtag gacaaaagat aaggaataca atacaaaaaa gatctttatt tattctttcc ttcctttatc cctattcata cagaattcct catgaactaa tgccaaattc tttccattta ttaattgcta caacgagtga tttattccaa tattagtta ttaccgaaca aagcaaaatt attack aaggatgaga tcaattcgga agcgcttttt tgttattcta gcagacgtaa ttgctttggt ctaattttgg 3480 gctttccaat caattttatc ttatctaatt ctatctatgc ccagaggatg ataccgaaac gaaacaatcc tttccttttt tctgatcata gaggagccgt atgaagctaa ggtttcatgt acggttttgg aatagcggtg agaactgtga tgttatcatc gactatgatt atctaatagt tcaagtacag ttgatatagt tgaagcacag tccaaatatg gtttttttgg atggaatatt tggcgtcagc ctataggttt tctagtttttt ctaatttctt ctttggcaga atgtgaaaga ttaccctttg atttaccaga agcagaagaa gaattagtag caggttatca aaccgaat tctggtatta aatatggttt attttatctt gtttcttacc taaatttatt agtttcctct ttatttgtaa ctgttctata cttaggcggg tggaattct ctattcccta tatatccttt tttggatttt tccaaatgaa taaataatt ggaattttgg aaatggtaat aggtatcttt 4020 attacatta ctaaagctta tttatttctc ttcatttcta tcacaataag atggacttta 4080 cccataatga gaatggatca gttattaaat cttggatgga aatttcttt acctatttct 4140 ctgggcaatc tcttattaac aacttcttcc caactagttt cactataaat aagaatacaa 4200 taacagtaag aatattttca acacaaacgt tctctcaaac aagagaaaga aacatacctt 4260 tttcatatat agatttagaa tatgttccct atgctaactg ggttcattac ttatggtcaa 4320 caaacaatac gcgccgcaag atacattggt caaagtttca taattacctt atcccacaca 4380 aatcgtttac ctataacgat tcactaccct tatgaaaaat caattacatt ggagcgtttc 4440 ctggggcgaa tacactttga atttgataaa tgcattgctt gtgaagtatg tgttcgcgta 4500 tgcccgatag atctaccctt tgtggattgg agatttgaaa aggatattaa aagaaaacaa 4560 ttgcttaatt atagtattga tttcggagtt tgtatatttt gcggtaactg tgttgaatac 4620 tgtcccacaa gttgtttatc aacgactgaa gaatatgaac tttctactta tgatcgtcat 4680 gaattgaatt acaatcagat tgctttgagt cggttaccat aatgggagat tacacaattc 4740 aaacaattag aaatttgcct caaagtaaaa tagacgaaga aaaatcttgg aattcaagaa 4800 cgattacaga ttactaggta ttaggatttt ttttattaga aaaatccatt tttactaact 4860 ctaacgaaaa agaataacta ctgattaaca acttatatgt atatacaaaa aaatatccta 4920 ataccttttt ccttccttga atcttttagt tttagtcagt tcatgaaaaa ttttatacta 4980 gaaatttctt cttatccata atggatttac ctggaccaat acatgagatt cttgtgctat 5040 ttgggggatt tggtcttcta ctaggaggtc taggagtagt attacttacc aacccaattt 5100 attctgcctt ttcgctggga ttagttcttg tttgtatatc cttattctat tttttattaa 5160 attcctactt tgtagctatc gcacaacttc ttatttatgt gggagccata aatgtcttga 5220 tcatatttgt tgtaatgttt gtaaacggct cagagtggtc taaagataag aattattgga 5280 ctattggaga tgggtttact ttactccttt gtataactat tcctttttca ctaatgacta 5340 ctatcccaga tacgtcgtgg catggaattc tttggactac aagatcaaac caaatagtag 5400 aacaggggtct cataataac gttcaacaa ttgggattca tttagcaacc gatttttatc 5460 ttccatttga actcattcc ctaatcttg tagttcttt ataggtaat tactatggct 5520 cgacaatag aaatacttag atgagtcaa attcttaga atttcaata taaataat 5580 aactaagaa tcacaattt gatttagtaa aacccatcta ctgccaatac aacaaatacc 5640 ttctttctc tttgttgcg taattgttct attcttagtta attgaatcag ttcattctt 5700 gtcctcatat tgaatgaat cgagattgat aaggaggtag ttaatgatgt tgagcatgt 5760 actttttg agtgtctatt tattttcgat tggtatctat ggattgatca caagccaaaa 5820 catggttaga gctctatat gtcttgaact tatactaat tcattaatc taaatctcgt 5880 aacattttct gatctatttg atagtcgcca attaaagga gatatttcg caatttgtt 5940 atagcccttg cggctgctga agcagctatt ggactatcca ttctttctc catccatcgt 6000 aacaggaaat caaccgtat caatcaatcc aatttttga atattagac atagaatccc 6060 ctaaaacaag gggcatatat aacattaga tgaatagaa tctaatctta 6120 ttttcttatt agtgtttaat aatatccttt ttttgagtag gttatttcag agtattgttt 6180 tttacttatt gaatattgca ttttgcaat tcattgatat tgcaatttga atattgcaat 6240 aatttatatt gaaaagatga tagccaattt attggctaat tcgaattagt atgtagaatt 6300 tgtataatta taactgttga agccttaaat tcaagtctct tggctctttt cacgctttct 6360 cacaaacaga ttacgaaata tattgcatta tttgttaaag tttggataaa ctattgcttc 6420 gtctggtgtc tacaatacat ctaatttata tagtactaat ttcattttta ccagatcgaa 6480 aattttatg ttgaaaagga aaatttagag atccaatgtc acattctgta aaaatttatg 6540 atacatgtat aggatgcact caatgtgtac gagcttgccc aacagatgta ttagaaatga 6600 taccttggga tggatgtaaa gccaagcaaa ttgcttccgc gccgagaacc gaagattgtg 6660 tgggttgtaa gagatgcgaa tccgcctgtc caacggattt ttaagtgtcc gcttttattt 6720 agggcctgaa acaacccgca gcatggctct atcttattga tacgttacaa aaaaactcca 6780 cttgaatcgt ctgattcctc tttaccgaag aagcctgtgc tcgaaatat cgagcatggg 6840 cttttctgat caaaacgtat cttgtattta ttactttatc atgagttatt ttccttggtt 6900 aacaatactt gttgttttgc cgatatttgc aggttcatta atttctttt tacctcataa 6960 aggaaataaa atcatatact atagctattt gtttattaga attccttcta atgacttatg 7020 cattctgtta tcatttccaa ttggaggatc ctttaatcca attaaaggag gattctaaat 7080 ggatagatgt ttcgatttc cactggagat tgggaatcga tggactttca ttaggatcta 7140 ttttattgac aagatttatc actactttag ctactttagc ggcttggccg gttactcgga 7200 attcgcaatt attctatttc ctgatgctag caatgtatag tggtcaaata ggattatttt 7260 cttcatgaga ccttttactt tttatcatg tgggagttag attaattcc tgtttactta 7320 cttttatcca tgtggaggga aagaggcgta tgtattcagc taccaagttt attttgtata 7380 ctgcaggcgg ttccatttt ttcttaattg gagttctggg tatgggatta tatggttcca 7440 atgaacccgg attagattta gaaagattga ttaatcaatc ataccctaca acattggaaa 7500 tactactgta ttttggcttc cttattgctt atgctgtcaa attgccgatt atacctttac 7560 atacgtggtt accagatacc catggggaag cgcattacag tacatgtatg cttttagccg 7620 gaattctatt aaagatggga gcatacggat tgattcgggt caatatggaa ttgttaccgc 7680 atgctcatta tctattttcc ccttggttgg tataatagg agcggtgcaa ataatctatg 7740 cagcttcaac ttctcttggt caacgaaatt tcaaaaaaag atagcctac tcctccgtat 7800 ctcacatggg tttcataatt ataggaattg gttccataac caacattgga ctaaatggag 7860 ctattttaca aattatct catggattta tcggtgctac actttttttc ttggcggggaa 7920 cggacttgtg atagaatgcg tcttgttat ctcgaagaac tggagggaat atctatccca 7980 atgccaaaaa ttttaccat gtttagtagc ttttcagtgg cttctcttgc cttgccggga 8040 atgagcggtt ttgttgcaga attagtagta ttttttggac taattactag tcctaaattt 8100 atgttaatgc caaaaatgct aattactttt gtaatggcaa taggaatgat attaactcct 8160 atttatttat tatctatgtt acgccagatg ttctatggat acaagctatt tcatgttcca 8220 atcaaaaatt ttgtagattc tggaccacgg gaactctttc ttttaatctg tatcttttta 8280 ccagtaatag atctatctcg aattttcaga attagatcta tctcgaattt ttcagaatta 8340 gatctttttc gtatctttca gaattagcta tttcatgttc catctttttc agaattagat 8400 ctatctcgaa tttttgagaa ccccttgaac gtcttttcaa agggttctca aatcaaacta 8460 aaaaggaaaa aaaccgaagg tttatgtta tgtaattat tagatggtaa tgtaaatgaa 8520 ccgtaactat gtaaacctat tcctaacaga ttgataccaa aatagcagat ccaaattata 8580 agaaatccta tcgaagctac aagtgcggaa ttcgtaccct tccaatttgg attttttcta 8640 ctatgtaaat atattgcaaa tatggtccag gtaataaatg cccaagtttc cttaggatcc 8700 caattccaat aggatcccca tgcctcatta gcccatactg ctccacaaag aatacccacg 8760 gttaaaaagag taaaccctag actaatgaca cgataactcc aagaatccaa acgctcagtt 8820 aattgatatt tgtaataatt tggaaatacg ggaaaagagg tgttttttaa agcacttctt 8880 tttgcatata aatattcaat ctcactaaag aaaaatgttt taagaaaaac attttcttt 8940 agtgaaaaga aatcgaaatt ctttcgaaat ctaatgatta gaagagcggc ggataataag 9000 gatccacaca aaagagttgc atagcttagt aacatcatac tgacatgcat cattaaccac 9060 tgagattgta gagcaggtac tagtattgtg gattgatgca tttcagttaa aagacccgac 9120 gtggcaaagc cttgcgttaa aatagtactt ggcgtagtta ttgtgcttaa atcatttttc 9180 gagttctgta tcttaggaat agtatgaaga atatacagag tccatgaaag gaagatcaat 9240 gactcatata aattacttaa tggaaaatgt cccgaagaaa cccaacgaga gactaaaaat 9300 cctgttatag agaaaaaagt agctatcatt cctttttctg acgaatcacg taatccccta 9360 agttcacgaa ctaataaggt tatcaaatga atcgtaatca caattgaaat ggttgagaaa 9420 gagatatgag ttagtatatg ttctaaagtt gcaaatagca taacgataag gtcccattac 9480 aaaattggaa atttcgaatt gaatccattt tctaattttt gtattctttt tcgagaatgc 9540 cgccactcgg attcgaaccg agatgcttga gcactgcttc ctaagagcag cgtgtctacc 9600 aatttcacca tggcggctaa tttaaaataa tagttaactt aaaagaataa tagttatttt 9660 atcgtgaatc gtcgagactg ggagaagcca tagaatttag gaacattaga agttcatcat 9720 tagagttca tcattaacta caatgaatg cattttgtat tttagaaaaa tgatagaatg 9780 aaagccttta cactcttatt atatgatgt accagtccta aacccatta tatgggaatt 9840 ttggataga ttaggtggag gttgtaagtc ctattgcaag atagttact ttttaata 9900 gatcctcgt ttttatgaga attcttatt aaaaaaagt tctttgatag gaaaagaata 9960 ctgaggacac atattaccaa aaacttttgt tctatataat gatatggag ggattcgttc 10020 taagaatatt gtaccgagga attcgacaca taaaagtaca tttaatt atccgaatt 10080 attcattcat attaataat tggaatttct tttgatagt tcattcaga ttttcaa 10140 atcttattat ttgttgctt gttgcccaga aaaacctttt gttttggat gctcgttgcc 10200 cctagaaaat gatcttgatt ttgctaaga atagattgt actaggaaa ataagtctt 10260 ttttttccaa agattttac gatacgctt tttgacatc gaagtacgtt ttttggac 10320 tgccattcaa aaagggaatt acttttttct agttgtatgt gaagacac tattgccaca 10380 aatcaatcct tctttctgtt tttcttagt atttatactt agatactgaa agatacttaa 10440 attctaaatt cttctttagt tcattttgtc tatgtgt agacaagg tttttaga 10500 tttttattta aatcaatttc tatatcaaat atactccata tataaatata tggtatgggg 10560 cataagccct catatagag ggggagaata aaattcatt caatagtta 10620 attacktca gaaggctatt ccatattga attccaataa aaaaaatac ttagtctctt 10680 aaacaagaca ttctaaaact tagagaattc tagtcattag gatttcctttt tatatgaat 10740 aagcaatatt cgagaatttc ctataaatat aggttctt tggaattca tcaatcaat 10800 acgaaaaaag agagctcttt tattttaca aaaagaata aaaatgat tagaagta 10860 aattattcaa tctttttttg tattttaata atatttttt tttactaata actagatacc 10920 gaaattcttt gattcacttt ttgaatttaa gtaactaaac ccattctaaa ttttggaata 10980 ttttaatgag agaaattaga aaaattcata attccagtat ttttattt tcttatttg 11040 tttttaat tcctttagaa agagataaga ataggttttgg tgaatcggaa acacatttat 11100 tttatagaaa aattgaacta taaatttaaa ctaaaaattg ctatttcttt tcttatggaa 11160 catacatatc aatatgcctg ggtaattcct cttctcccac ttccagttat tatgtcaatg 11220 ggattggac ttttttat tcctacagca acaaaaaatc ttcgtcggat atgggcttttt 11280 cctagtattt tactcttaag tatagctatg gtattctcac ttcacctgtc tattcaacaa 11340 ataaatggaa gttctatcta tcaatatcta tggtcttgga ccatcaataa tgatttttcc 11400 ttagaatttg gatacttggt cgaccccctt acgtctatta tgttaatact aattactact 11460 gtaggaatct tagttcttat ttatagtgac gattatatgt ctcacgatga aggatatttg 11520 agattttttg tttatataag ttttttaat acttccatgt taggattggt tactagttcc 11580 aatttgatac aaatttatt ttttgggaa cttgtcggaa tgtgttccta tttattgata 11640 ggctttggt ttacgcggcc aattgcagcg agtgcttgtc aaaaagcttt tgtaactaat 11700 cgtgtagggg attttggtct gttattagga attttaggtt ttttttggat aacgggtagt 11760 ttggagtttc gggatttgtt caaaatagct aataactgga ttcctaataa tgggattaat 11820 tccttactta ctactttgtg tgcttttta ttattccttg gtgcagttgc aaaatctgca 11880 caatttcctc ttcacgtatg gttacctgat gctatggaag gacccactcc tatttcggct 11940 cttatacacg cagcaactat ggttgctgcg gggatttttc ttctagctag acttcttcct 12000 cttttcatat ccctaccctg gataatgagt ttcatttctt tataggtac aataacactc 12060 ttcttaggag ccactttagc tcttgctcag agagatatta aaagaagctt agcctattct 12120 acaatgtctc aattgggtta tatgatgtta gctctaggta taggttctta tcaagctgct 12180 ttattccatt tgatcactca tgcttattcg aaagctttat tgttcttagg atccggatcc 12240 gttattcatt caatggaacc tcttgttgga tattcaccag ataaaagtca gaatatggtt 12300 cttatgggtg gtttaagaaa attacgttcca attacaagaa ctactttttt atgtggtaca 12360 ctttctcttt gtggtattcc acctcttgct tgcttctggt ccaaagatga aatccttagt 12420 aatagttggt tgtattcacc cttttttgga ataatagcct cttttactgc aggattaact 12480 gcattttata tgtttcggat atatttactt acttttgatg gcattttgcg tgttcatttt 12540 caaaattaca gtagtactaa agaaggttcg ttgtattca tatccttatg gggaaaaagt 12600 atatccaaag gagtcaatag ggattttgtt ttatcaacaa tgagagtgg agtttttttt 12660 tttcacaaa ataccaaa aattcctgct atacaagaa atagatagg atcctttagt 12720 actccctttg gggctaaaaa tactttgtc tatcctcatg aaacgggaaa tactatgcta 12780 tttcctctc ttatattact actttttact tgttcattg gatccatagg aatccatttt 12840 gatatggag windowgaata gatatattg gagttaacca tattacaa gtggctaact 12900 ccttcaataa acttgttcca ggaaaatttct aattctcca taaattcata tgaatttctc 12960 actaatgcaa ttctctctgt aagtttagca atttttggtc tattcatagc atatatcttt 13020 tatggatctg cttattcttt tttcagaat ttgaatttc aaaattccct tgtaaaaag 13080 aaatccaaaaa agagcttttt ggatgaagta aaaaaaaga tatacagctg gtcatataat 13140 cgtggttata tagatttt ctatactagg gttttttacc taggtataag agattagcc 13200 gaactaacgc atttttttga taaaggtgtc attgatggaa ttaccaatgg agtaggtctt 13260 gctggttttt gtataggaga agaaatcaaa tatgtagggg gagggcgaat atcgtcttat 13320 ctattctttt tttatgtta tgtatccttg ttcttattct ttatccatg aaaatggatt 13380 attccatgaa ttcctcaaaa cgaggctcat caaaatgcaa aatctaagac tactataaga 13440 ctactataata aataagaaaa aaatgagaac gattaaatta cttctcccga atatccaact 13500 gacttattaa tttcttataa cgtactctat ttttctttgc caaataagcc agcaaacgtt 13560 gacgttttcc caaaagtctt cgtagacctc tttccgatga aaaatctttt ttgtgtaatt 13620 ctaaatgtga agcaagtctc cgtatcttat tggtgaaact gaatacttga aattcaacag 13680 aacccctgtt ttcttgtttt tcttctttaa ccataaatca acaaattttt ctacctcctt 13740 tctttttcat gtatttttct gatcaggaaa aataaaaaat tatgtcagtt atttttaagt 13800 tattctaatc tcgtacacac aaaaatttgc aattattcat ctactgctgg aatctggaat 13860 ttggatttat tttatcgatg caaattggat ttggatagaa gggtacattc ttttatttta 13920 gatagaagaa acatttcttc tatctaaaat aaaagaattt tgctgattta tttattgcta 13980 tatccaattt atagaattga tataccattt aattgatatc atttagcaaa atgaaacaca 14040 gcatatgcat ccatctttgg gctcgagaat ttcacgggat agatatag tataagaaat 14100 aggctattaa gtaactctaa atgaattgtg gatacatctg tatccttaac atactgaaac 14160 gactgccatt attcgtatca aaccaatagc gattcataaa agctaaatct tgtaatcaat 14220 ggtgggccaa taatgaattt ttttgcatat gtattaagac gcttggtctg atttcgaaat 14280 tgtccagagt tttttatgtt gttttcattg caaaatgatg gatctccttc cgtaactttc 14340 caattacgag tacgagaatt gaaagacatg aaaattctca attctctacg gcgtctagga 14400 gatggatcga atattttcag gaacaagaaa atcagaagaa tcttttctc tattcactac 14460 cattccgcgt cttcgacttc tattagtttc ttttcttctt taatgcaata gctatagttt 14520 gatatagaat ccatttctca aagtaatgga aaccattctc ttataggaaa tggttcgaaa 14580 atcgctattc caccttttag gtttaggtat cgtgaaaagt gatacctgtg aagatcgtgc 14640 atttcagtca cattcagatc cgtttttcga gtccatgata taaccaaatt ggatggatct 14700 tccaccccgtt tagctaagaa agaatagatg cagaggtgga taatagatcg atatagaat 14760 catgagctgc cccataatga aaccgccagt agtcgcgaat atctccttct tccctaaccc 14820 aagattggag aaagaagatc taagagggac ctatggagaa tgtggtcaga aatccataat 14880 14940 ctacttaagt agaaaagatt tgaaaatcat ggcatgggtc tccttttttt ctttctttag 15000 agttttctat atgcacaatt tctcgatgtt tcgatgagaa tttcttgact ttccatatat 15060 agaaagagat agactataaa tgacatctct tatgtcaata agaccaaagg aatggatatt 15120 15180 atgaaatgag gcatggaacg gaggccactac gaagaaattc cgggagttac gaagaagct 15240 tcggactcat attgttcacg ggttgagagc gggagttgaa ctctaggagg tcgaatctcc 15300 ccttgttcct footgctca gtggtaggc ggtcggctgt taactgactg gtcgtaggtt 15360 cgaatcctac ttggggagat ttgattcatt ctttaatgta agaataaaga attgaattaa 15420 aaggcttgct ttgaccctta ggagtaggta acccgttcgc tatccttgtt tctattgcat 15480 tctgtctcat cgtatcacat tctgttctac gattccactt cgacaaaagg aaagagcata 15540 cccaagttca atagctttac gtccgctatt ccgatcatga ttttcctacc ctcagggaga 15600 aagtaaaggt ccttccccct ttggaaggct gtgggcgagg agggattcga accccccgaca 15660 ccgtggttcg tagccacgtg ctctaatcct ctgagctaca ggcccacccc gtctccactg 15720 gatctcttcc cggggatacc ccccaaaagg aaccttcttc tcctcagcca tttcatttcg 15780 ggttaagaag atgggaaagc gcctttctct ctataagaac agtgcgttct gaggtgtgaa 15840 gtgggagaga ggggatgatt gaggttttga ataagacgac ctttgcgttt tggatttgga 15900 tctttttcgt atttcaaaat agtgaaaaag tcaaataaga ggtgttaagc tttttatcat 15960 tctggcatcg agctattttg ccgcaggacc tccctacag tatcgtcacc gcagtagagt 16020 ttaaccacca aattcgggat ggattggtgt ggttcctcta cgcctaggac accagaatat 16080 cgaaccatga acgagaaaag gcatgagaga aatattggct agtaattgtg aagtcccaat 16140 tcttaactgg aagggacacc aaaggactct gccctccctc tctatttatc caagagatgg 16200 aagggcagag cttttttttg gttttttcat cttttcttttt catcaaagag ttgaacaatg 16260 aagatagatg gcaagtgcct gatcgatttg atcaggtcgt gtaggaacaa ggttcaaatc 16320 gttcgttcgt taggatgcct cagctgcata catcactgca cttccacttg acacctattt 16380 aaacggctcg tctcgccgct accttatcct atttccatac ttctgtcgct ccatccccgt 16440 atgggtggag aacccgtcgc tgtctcggct gtgctaccgg aggctctagg gaagtcggag 16500 gagagagcac tcatcttggg gtgggcttac tacttatatg ctttcagcag ttatcctctc 16560 cacacttggc tacccagcgt ttaccgtagg cacgataact ggtacaccag aggtgcgtcc 16620 ttcccggtcc tctcgtacta gggaaaggtc ctctcaatgc tctaacgccc acaccggata 16680 tggaccgaac tgtctcacga cgttctgaac ccagctcacg taccgcatta atgggcgaac 16740 agcccaaccc ttggaaccac ctacagctcc aggtggcgaa gagccgacat cgaggtgcca 16800 aaccttcccg tcgatgtgga ctcttgggga agatcagcct gttatcccta gagtaacttt 16860 tatccgttga gcgacggccc ttccactcgg caccgtcgga tcactaaggc cgactttcgt 16920 ctctgctcga cgggtgagtc ttgcagtcaa gctcccttct gcctttgcac tcgaggacca 16980 atgtccgtct ggcccgagga aacctttgca cgcctccgtt accttttggg aggcctacgc 17040 cccatagaaa ctgtctacct gagactgtcc cttggcccgc gggtctgaca caaggttaga 17100 atccgagctc ttccagagtg gtatctcact gatggctcgg gcccccccgg aagggggcct 17160 tcttcgcctt ccacctaagc tgcgcaggaa aagcccaaag ccaatcccag ggaacagtaa 17220 agcttcatag ggtctttctg tccaggtgca ggtagtccgc atcttcacag acatgtctat 17280 ttcaccgagc ctctctccga gacagtgccc agatcgttac gcctttcgtg cgggtcggaa 17340 cttacccgac aaagtaattc gctaccttag gaccgttata gttacggccg ccgttcaccg 17400 gggcttcggt cgccggcttc cctgtcatca gttcaccaac ttccttgacc ttccggcact 17460 gggcaggcgt cagcccccat acatggtctt acgactttgc ggagacctgt gtttttggta 17520 aacagtcgcc cgggcctggt cactgcgacc cccttttgcg aggggcacc ccttctcccg 17580 aagttacggg gctattttgc cgagttcctt aagagagtt gtctcgcgcc cctaggtatt 17640 ctctacctac ccacctgtgt cggtttcggg tacaggtacc cttttgttga aggtcgttcg 17700 agcttttcct gggagtatgg catgggttac atacttcagc gccgtagcgc ctggtatgag 17760 cctcgtggag aagcaatggc tagtccacgg ggctcatact tcagcgctgc agcgcttggt 17820 actcggacct cggctcgagg cattttctct accccttctt accctgaaaa agcagggtca 17880 ccttgtgtcc ttaaacctat aaccatcttt cggctaacct agcctctcc gtccctccgt 17940 accaacaagg ggtagtagcag gaatattgac ctgttgtcca tcgactacgc ctttcggcct 18000 gatcttaggc cctgactcac cctccgtgga cgaaccttgc ggaggaaacc ttgggttttc 18060 ggggcattgg attctcacca atgttttcgt tactcaagcc gacattctcg cttccgcttc 18120 gtcgaccccc gctttcgcgt ttgcttccct ctaaggcgga acgctcccct accgatgcat 18180 tttgacatcc cacagcttcg gcagatcgct tagccccgtt catcttcagc gcaagggcgc 18240 tcgatcagtg agctattacg cactctttaa agggtggctg cttctaggca aacctcctgg 18300 ctgtctttgc acccccacct cctttatcac tgagcggtca tttaggggcc ttagctggtg 18360 atccgggctg tttccctctc gacgatgaag cttatccccc atcgtctcac tggccgacct 18420 tgacccctgt tatttttggg tcatatctag tattcagagt ttgcctcgat ttggtaccgc 18480 tcgcgcagcc cgcaccgaaa cagtgcttta cccctagatg tccagtcaac tgctgcgcct 18540 caacgcattt cggggagaac cagctagctc tgggttcgag tggcatttca cccctaacca 18600 caactcatcc gctgattctt caacatcagt cggttcggac ctctgcttag tttcatccaa 18660 gcttcatcct ggtcatggat agatcaccca ggttcgggtc cataagcagt gacaatcgcc 18720 ctatgaagac tcgctttcgc tacggctccg gtgggttcca ttcccttaac caagccactg 18780 cctatgagtc gccggctcat tcttcaacag gcacgcggtc agagatcact ttcccctccc 18840 actgcttggg agctcagcac ggtttcacgt tctatttcac tacccactgg gggttctttt 18900 cacctttccc tcacggtact acttcgctat cggtcaccca ggagtattta gccttgcaag 18960 gtggtccttg ctgattcaca cgggattcca cgtgccccat gctactcggg tcagagcgta 19020 agctagtgat gctttcggct actggacttt agccatctag ggtgcggcac tcaaccgctt 19080 cgcctagcag cacaacgctt gtattgctct cccacaaccc cgttttcacg gtttaggctg 19140 ctcccatttc gctcgccgct actacgggaa tcgcttttgc tttcttttcc tctggctact 19200 aagatgtttc agttcgccag gttgtctctt gcctgctcat ggattcagca ggcagttcaa 19260 aaggttgacc tatttgggaa tctccggatc tatgcttatt ttcaactccc cgaagcattt 19320 cgtcgcttgc tacgcccttc ctcgtctctg ggtgcctagg tatccaccgc aagcctttcc 19380 tcttttgaac ctcgccatta acgttaaggc tatgccatcc taaggtgcta ctaaatggaa 19440 ggatcttatc aacgtccatg aatgcgaaat catagatcga actgacgaat tggcaacctt 19500 cggtgctatc atagtatccg ctaagttcac gggctggaga taagcggact cgaaccgctg 19560 acatccgcca cagggtaaac caccgcctct caggcctccc cgacgggttc taccatagag 19620 gacaacgata ggcaataact cccccccgaa cacagcttac aactttcatc gtactgtgct 19680 ctccaaagag caactcttct caaaatccca aaacaaaagg tgctgagttg gaatcccatt 19740 ctaaggattc ttgtggttcc ggggaatcca gctacaggag aaccaggaac ggggagctct 19800 cccctttttc cgcccgactc tttgatctta acttaagaat gctggtttta agaacgagtg 19860 attgcccttc tccgaccctt actgcccaac cggagagcgg acggctaatg tgttccactt 19920 attgaacagg atctatggtc ggtccgtgac ccctggacgc cgaaggcgtc cttggggtga 19980 tctcgtagtt cctacggggt ggagacaatg gggtcggtcc atggattttc cttccttttg 20040 ccacatttcg ctcaaagggt tgaagggaga tagtgcatca agctattcgc aagggccaac 20100 ttgatcctct tccccaggga tcccagatga gggaagccta ggagagccgc cgactccaac 20160 tatcgtccat gtacgatcca tactagatct gaccaactgc ccatcctacc tcctctacct 20220 ttttgacagc ccatcttttt gtctcagtag agtctttcag tggcatgttt cagtcctctt 20280 ccccattact tagaaaaagt gagccaccgg ttcaggtaca agatactatc attaccgcct 20340 ggacaattag acagccaacc cgtaatcgca acgacccaat tgcaagagcg gagctctacc 20400 aactgagcta tatccccccc gagccaagtg gagtatgcat gaaagagtca gatgcttctt 20460 ctattctttt ccctggcgca gctggggccat cctggacttg aaccagagac ctcgcccgtg 20520 aagtaaatca tcgcccctac gatccaacca attgggagag aatcaataga ctcctttcg 20580 ggagcgattc atccttcccg aacgcagcat acaactcccc gttgtactgc gctcttcaag 20640 tgtgcttctt cccccttctc ccccttacca cggcaagtcc ttgggaaata actccgatgg 20700 gcagaaaaaa gaaggggtta agagaccctc ctggcccaac cctagacact ctaagatcct 20760 tttcaacc tgctctgctc ccattcgag tcaagagata gataatagc cacatcccat 20820 tgcactgatc gggggcgctc gtagtgactt agggggtcga agaccagaa gtggcttatt 20880 tataccaagc attcctctta tggctagatc caatctcctg gtccctgcag aaggaaaaa 20940 gatttcacg ttctccttt caggaaggga ggattaggga agtcctattg attactgctt 21000 tctccagacc gccgggaaa gcatgaaaaa aaggctcgaa tggtacgatc cctccgtcac 21060 cccagaatga aagggtgat ctcgtagttc ttggtctgtg aagatgcgtt gttaggtgct 21120 ccattttccc attgaggacg aacctcacc tgtgctcgag agatagctct ccatacactg 21180 atagggatg tatggattct cgagagaga ggaggccgcgg tggccccccc ccggaccgcc 21240 cggatcccac gagtgaatag aaagttcgat ctacatggga tctcacctga atcgccccat 21300 ctatcctcct gaggagagt ttgttttggtt tcaactccg attchaacag gaggagtacg 21360 ccatgctaat gtgccttgga tgatccacat cttcggtca ggcgctgatg agcacattga 21420 actatccatg tggctgagag cccctcacagc ccaggcacaa cgacgcaatt atcaggggcg 21480 cgctctacca ctgagctaat agcccgtcgc gcgggcctcc caaagggagg cctgctacgc 21540 caaaagcgag aaaaactcca tccctttcct tttgacatcc ccatgccgcc acacgggggg 21600 acatggggac gtcaaaaagg ggatcctatc actatcaact aatttgttcc gacctaggat 21660 aataagctca tgagcttggt cttacttcac cttaaacgaa agaagacttc catatccaag 21720 tttagctcag acgtagctgc cttctttttg ggcgtgaagc agtgtcaaac caaaataccc 21780 aataagcata agcattagct ctccctgaaa aggaggtgat ccagccgcac cttccagtac 21840 ggctaccttg ttacgacttc actccagtcg caagcctagc cttaggcatc cccctcctta 21900 cggttaaggg taatgacttc aaacctggcc agctcctata gtgtgacggg cggtgtgtac 21960 aaggcccggg aacggattca ccgccgtatg gctgaccggc gattactagc gattcctgct 22020 tcatgcaggc gagttgcagc ctgcaatccg aactgaggac gggtttttgg agttagctca 22080 cccgcgag atcgcgaccc tttgtcccgc ccattgtagc acgtgtgtcg cccagggcat 22140 aaggggcatg atgacttggc ctcatcctct ccttcctccg gcttaacacc ggcggtctgt 22200 tcagggttcc aaactcatag tggcaactaa acacgagggt tgcgctcgtt gcgagactta 22260 acccaacacc ttacggcacg agctgacgac agccatgcac cacctgtgtc cgcgttcccg 22320 agggcacccc tctctttcaa gaggattcgc ggcatgtcaa gccctggtaa ggttcttcgc 22380 tttgcatcga attaaaccac atgctccacc gcttgtgcgg gcccccgtca attcctttga 22440 gtttcattct tgcgaacgta ctccccaggc gggatactta acgcgttagc tacagcactg 22500 cacgggtcga gtcgcacagc acctagtatc catcgtttac ggctaggact actggggtct 22560 ctaatcccat ttgctcccct agctttcgtc tctcagtgtc agtgtcggcc cagcagagtg 22620 ctttcgccgt tggtgttctt tccgatctca atgcatttca ccgctccacc ggaaattccc 22680 tctgccccta ccgtactcca gcttggtagt ttccaccgcc tgtccagggt tgagccctgg 22740 gatttgacgg cggacttgaa aagccaccta cagacgcttt acgcccaatc attccggata 22800 acgcttgcat cctctgtctt accgcggctg ctggcacaga gttagccgat gcttattcct 22860 cagataccgt cattgtttct tctccgagaa aagaagttga cg 22902 <210> 25 <211> 1029 <212> DNA <213> Zea mays <400> 25 atggatttac ctggaccaat acatgagatt cttgtgctat ttgggggatt tggtcttcta 60 ctaggaggtc taggagtagt attacttacc aacccaattt attctgcctt ttcgctggga 120 ttagttcttg tttgtatatc cttattctat tttttattaa attcctactt tgtagctatc 180 gcacaacttc ttatttatgt gggagccata aatgtcttga tcatatttgt tgtaatgttt 240 gtaaacggct cagagtggtc taaagataag aattattgga ctattggaga tgggtttact 300 ttactccttt gtataactat tcctttttca ctaatgacta ctatcccaga tacgtcgtgg 360 catggaattc tttggactac aagatcaaac caaatagtag aacagggtct cataaataac 420 gttcaacaaa ttgggattca tttagcaacc gatttttatc ttccatttga actcatttcc 480 ctaattcttg tagtttcttt aataggaatc cattttgata atggagtaaa agataataga 540 atattggagt taaccatatt atcaaagtgg ctaactcctt caataaactt gttccaggaa 600 aattctaatt cttccataaa ttcatatgaa tttctcacta atgcaatttc ttcttcaagc 660 aatttctaatt cttccataaa ttcatatgaa tttctcacta atgcaatttc ttcttcaagc 660 tcccttctgc ctttgcactc gaggaccaat gtccgtctgg cccgaggaaa cctttgcacg 720 tcccttctgc ctttgcactc gaggaccaat gtccgtctgg cccgaggaaa cctttgcacg 720 cctccgttac cttttgggag gcctacgccc catagaaact gtctacctga gactgtccct 780 cctccgttac cttttgggag gcctacgccc catagaaact gtctacctga gactgtccct 780 tggcccgcgg gtctgacaca aggtaccctt ttgttgaagg tcgttcgagc ttttcctggg 840 tggcccgcgg gtctgacaca aggtaccctt ttgttgaagg tcgttcgagc ttttcctggg 840 agtatggcat gggttacata cttcagcgcc gtagcgcctg gtatgagcct cgtggagaag 900 agtatggcat gggttacata cttcagcgcc gtagcgcctg gtatgagcct cgtggagaag 900 caatggctag tccacggggc tcatacttca gcgctgcagc gcttggtact cggacctcgg 960 caatggctag tccacggggc tcatacttca gcgctgcagc gcttggtact cggacctcgg 960 ctcgaggcat tttctctacc ccttcttacc ctgaaaaagc agggtcacct tgtgtcctta 1020 ctcgaggcat tttctctacc ccttcttacc ctgaaaaagc agggtcacct tgtgtcctta 1020 aacctataa 1029 aacctataa 1029 <210> 26 <211> 342 <212> PRT <213> Zea mays <400> 26 Met Asp Leu Pro Gly Pro Ile His Glu Ile Leu Val Leu Phe Gly Gly Met Asp Leu Pro Gly Pro Ile His Glu Ile Leu Val Leu Phe Gly Gly 1 5 10 15 Phe Gly Leu Leu Leu Gly Gly Leu Gly Val Val Leu Leu Thr Asn Pro Phe Gly Leu Leu Leu Gly Gly Leu Gly Val Val Leu Leu Thr Asn Pro 20 25 30 Ile Tyr Ser Ala Phe Ser Leu Gly Leu Val Leu Val Cys Ile Ser Leu Ile Tyr Ser Ala Phe Ser Leu Gly Leu Val Leu Val Cys Ile Ser Leu 35 40 45 Phe Tyr Phe Leu Leu Asn Ser Tyr Phe Val Ala Ile Ala Gln Leu Leu 50 55 60 Ile Tyr Val Gly Ala Ile Asn Val Leu Ile Ile Phe Val Val Met Phe 65 70 75 80 Val Asn Gly Ser Glu Trp Ser Lys Asp Lys Asn Tyr Trp Thr Ile Gly 85 90 95 Asp Gly Phe Thr Leu Leu Leu Cys Ile Thr Ile Pro Phe Ser Leu Met 100 105 110 Thr Thr Ile Pro Asp Thr Ser Trp His Gly Ile Leu Trp Thr Thr Arg 115 120 125 Ser Asn Gln Ile Val Glu Gln Gly Leu Ile Asn Asn Val Gln Gln Ile 130 135 140 Gly Ile His Leu Ala Thr Asp Phe Tyr Leu Pro Phe Glu Leu Ile Ser 145 150 155 160 Leu Ile Leu Val Val Ser Leu Ile Gly Ile His Phe Asp Asn Gly Val 165 170 175 Lys Asp Asn Arg Ile Leu Glu Leu Thr Ile Leu Ser Lys Trp Leu Thr 180 185 190 Pro Ser Ile Asn Leu Phe Gln Glu Asn Ser Asn Ser Ser Ile Asn Ser 195 200 205 Tyr Glu Phe Leu Thr Asn Ala Ile Ser Ser Ser Ser Ser Leu Leu Pro 210 215 220 Leu His Ser Arg Thr Asn Val Arg Leu Ala Arg Gly Asn Leu Cys Thr 225 230 235 240 Pro Pro Leu Pro Phe Gly Arg Pro Thr Pro His Arg Asn Cys Leu Pro 245 250 255 Glu Thr Val Pro Trp Pro Ala Gly Leu Thr Gln Gly Thr Leu Leu Leu 260 265 270 Lys Val Val Arg Ala Phe Pro Gly Ser Met Ala Trp Val Thr Tyr Phe 275 280 285 Ser Ala Val Ala Pro Gly Met Ser Leu Val Glu Lys Gln Trp Leu Val 290 295 300 His Gly Ala His Thr Ser Ala Leu Gln Arg Leu Val Leu Gly Pro Arg 305 310 315 320 Leu Glu Ala Phe Ser Leu Pro Leu Leu Thr Leu Lys Lys Gln Gly His 325 330 335 Leu Val Ser Leu Asn Leu 340 <210> 27 <211> 21 <212> DNA <213> Maize <400> 27 atgacgagtt ttcacgtccg a 21 <210> 28 <211> 22 <212> DNA <213> Zea mays <400> 28 cgttcacgcg atttttcaag tg 22

Claims

1. A method of identifying a corn plant having increased disease resistance to northern corn blight, gray leaf spot, or southern corn rust, the method comprising: a. detecting any one of SEQ ID NOs: 4, 5, 8, and 9 in the corn plant genome; as well as b. Identifying the corn plant as having increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust.

2. A method of identifying a plant having increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust, the method comprising: a. detecting a polynucleotide in the genome of a corn plant, the polynucleotide encoding a polypeptide having an amino acid sequence as shown in SEQ ID NO: 2 or SEQ ID NO: 3; and b. Identifying the corn plant as having increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust.

3. A method for increasing disease resistance to northern leaf blight, gray leaf spot, or southern corn rust in a corn plant, the method comprising expressing in the corn plant a recombinant polynucleotide encoding any one of SEQ ID NO: 2 or SEQ ID NO: 3; wherein the corn plant expressing the recombinant polypeptide has increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust when compared to a control corn plant not containing the recombinant polynucleotide.

4. The method of claim 3, wherein the recombinant polynucleotide further comprises a heterologous promoter.

5. The method of claim 3, further comprising obtaining a progeny corn plant derived from a corn plant expressing the recombinant polynucleotide, wherein the progeny corn plant comprises the recombinant polynucleotide in its genome and exhibits increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust as compared to a control plant not comprising the recombinant polynucleotide.

6. The method of claim 3, wherein the recombinant polynucleotide is located at a position different from the position of the native sequence encoding SEQ ID NO: 2 or SEQ ID NO: 3 on maize chromosome 7.

7. A method for identifying a ZmMM1 gene variant that confers increased disease resistance to northern corn blight, gray leaf spot, or southern corn rust in a corn plant, the method comprising the steps of: a. combining one or more nucleotide sequences encoding one or more fragments of any one of SEQ ID NO: 2 or SEQ ID NO: 3 by gene shuffling to produce a ZmMM1 gene variant; and b. Identifying variants that exhibit increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust.

8. The method of claim 7, wherein the method further comprises the steps of: a. introducing a recombinant construct into a regenerative corn plant cell, the recombinant construct comprising the ZmMM1 gene variant identified by the method of claim 7; b. after step (a), regenerating a transgenic corn plant from the regenerable plant cell, wherein the transgenic corn plant comprises the recombinant DNA construct in its genome; and c. The transgenic corn plant of select (b), wherein the transgenic plant comprises the recombinant DNA construct and exhibits increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust when compared to a control plant not comprising the recombinant DNA construct.

9. A method for identifying an allelic variant of the ZmMM1 gene, wherein the allelic variant is associated with increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust, the method comprising the steps of: a. obtaining a population of corn plants, wherein the plants exhibit varying levels of disease resistance to northern leaf blight, gray leaf spot, or southern corn rust; b. assessing allelic variation with respect to a polynucleotide sequence encoding a protein comprising SEQ ID NO: 2 or SEQ ID NO: 3, or allelic variation in a genomic region that regulates expression of a polynucleotide encoding said protein; c. Associating allelic variation with increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust; and d. Identify allelic variants associated with increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust.

10. The method of claim 9, further comprising detecting the allelic variant associated with increased disease resistance and selecting a corn plant if the allelic variant is detected.

11. A method for introducing an allelic variant of the ZmMM1 gene, wherein the allelic variant is associated with increased disease resistance to northern leaf blight, gray leaf spot, or southern corn rust, the method comprising introducing a mutation in the endogenous ZmMM1 gene using a zinc finger nuclease, a transcription activator-like effector nuclease, a CRISPR / Cas system, or a meganuclease, such that the allelic variant comprises a polynucleotide sequence encoding SEQ ID NO: 2 or SEQ ID NO: 3.

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