Molecular marker related to corn plant dwarfing and application thereof

Abstract

The invention discloses a molecular marker related to corn plant dwarfing and application of the molecular marker. The molecular marker is characterized in that C at the 1099th site of a sequence as shown in SEQ ID NO.1 is mutated into T, or G at the 8405th site of the sequence as shown in SEQ ID NO.1 is mutated into A. For the variation, two pairs of molecular marker primers dgk2.1-F / R and dgk2.2-F / R are developed, and the wild type mutant and the dwarf type mutant can be accurately distinguished through sequencing after PCR (Polymerase Chain Reaction). The molecular marker is beneficial to auxiliary breeding of new corn dwarf varieties.

Description

Technical Field

[0001] The invention belongs to the technical field of molecular genetic markers, and in particular relates to a molecular marker related to dwarfing of corn plants and an application thereof. Background Art

[0002] Corn (Zea mays L.), as an important food, feed and industrial raw material, is the largest crop in my country and plays an important role in the national economy. Reasonable plant height can effectively reduce lodging, increase planting density, and contribute to high corn yield.

[0003] Plant diacylglycerol kinase (DGK) phosphorylates diacylglycerol (DAG) to produce phosphatidic acid (PA); DAG and PA, as endogenous lipid signaling molecules in plants, are widely involved in plant growth and development and stress response processes (Hong-Yan Yao, Hong-Wei Xue. Phosphatidic acid plays key roles regulating plant development and stress responses. J Integr Plant Biol. 2018 Sep; 60(9): 851-863.).

[0004] A large number of previous research results have shown that maize plant height is a complex quantitative trait controlled by a series of micro-effect polygenes. Although there are many QTL loci regulating maize plant height, there are still few genes that have been successfully cloned. Therefore, it is urgent to explore the key genes that regulate maize plant height traits that can be used in breeding. Summary of the invention

[0005] The purpose of the present invention is to provide a molecular marker related to corn plant dwarfing and its application, so as to provide a new breeding approach with greater application potential for corn plant height genetic improvement and dense-tolerant breeding.

[0006] The present invention is achieved through the following technical solutions:

[0007] The present invention provides a molecular marker associated with dwarfing of corn plants. The molecular marker is a mutation of C at position 1099 to T, or a mutation of G at position 8405 to A in the sequence of corn diacylglycerol kinase gene ZmDGK2 (GRMZM2G094452). The sequence of the corn diacylglycerol kinase gene ZmDGK2 is shown in SEQ ID NO.1.

[0008] The present invention also provides a pair of specific primers for detecting the above-mentioned molecular markers, and the specific primers include the primer pair dgk2.1-F / R for detecting the mutation site at position 1099:

[0009] dgk2.1-F: CGCTTTTAGGATGGTGTGGAT;

[0010] dgk2.1-R: AAGGTTGGCATATAAACGATGC;

[0011] and the primer pair dgk2.2-F / R for detecting the mutation site at position 8405;

[0012] dgk2.2-F: CCATGACATTTTCTCTCCTATGC;

[0013] dgk2.2-R: CTGACCCCGTTTTAGTGTACCA.

[0014] The present invention also provides the application of the above-mentioned molecular markers or specific primers in identifying or assisting in identifying the plant height trait of maize.

[0015] The present invention also provides the application of the above-mentioned molecular markers or specific primers in the genetic improvement of maize dwarf materials or the assisted breeding of new varieties.

[0016] The present invention also provides a method for identifying or assisting in identifying the plant height trait of maize, which includes the following steps: using the genomic DNA of the maize to be tested as a template, performing PCR amplification with the above-mentioned primer pair dgk2.1-F / R or dgk2.2-F / R to obtain an amplification product, sequencing the amplification product, and identifying the plant height of maize according to the base type of the mutation site.

[0017] Among them, for the type of base at the 369th bp of the amplification product obtained using the primer pair dgk2.1-F / R, if it is C, the maize material to be tested is of wild genotype; if it is T, the plant height of the maize material to be tested becomes shorter; for the type of base at the 248th bp of the amplification product obtained using the primer pair dgk2.2-F / R, if it is G, the maize material to be tested is of wild genotype; if it is A, the plant height of the maize material to be tested becomes shorter.

[0018] The sequence of the amplification product (wild genotype) obtained by dgk2.1-F / R is as follows (SEQ ID NO.2): CGCTTTTAGGATGGTGTGGATTCTGTTGTAAAGATAACATAATTCAGGACCACCCAATCAAAATATGCCTTGTATTCATGAATGGAGGGAGCAAACATTTGTAGCACCAGTGAGTTATCCCATTAAAACTATGAGAACGATTTTTCAGGATGGAAGGTGAGGCAGAGACCGTGGTTGGTTCTTGTTCTAAACCATGTGGGCCTCTGGAGGACTACTACATTCCAGATTACATTCTGAAGCCAGGTGCCCAACAAGTACTTGTTGATCATGCGGCACCCTGCCCCGTTGTAGTGTTCATCAACTCAAGATCTGGAGGCCAACTTGGAAGTAGTTTAATCAAAACATATCGTGAGCTTCTCAATGAAGCACAGGTACCTTTTGTCTTTCTGGAAATGCTAGACCATGGAAATATGCTTCTGGTCTTAATTGCATTTTGAATCTGTAATTTTTGTGAGAGCTGCTCAGTTACTTTCAGGTTTTTGATCTCTCAAAAGAGGCTCCAGATAAGGTATTGCATCGTTTATATGCCAACCTT

[0019] The sequence of the amplification product obtained by dgk2.1-F / R (homozygous mutant genotype) is as follows (SEQ ID NO.3): CGCTTTTAGGATGGTGTGGATTCTGTTGTAAAGATAACATAATTCAGGACCACCCAATCAAAATATGCCTTGTATTCATGAATGGAGGGAGCAAACATTTGTAGCACCAGTGAGTTATCCCATTAAAACTATGAGAACGATTTTTCAGGATGGAAGGTGAGGCAGAGACCGTGGTTGGTTCTTGTTCTAAACCATGTGGGCCTCTGGAGGACTACTACATTCCAGATTACATTCTGAAGCCAGGTGCCCAACAAGTACTTGTTGATCATGCGGCACCCTGCCCCGTTGTAGTGTTCATCAACTCAAGATCTGGAGGCCAACTTGGAAGTAGTTTAATCAAAACATATCGTGAGCTTCTCAATGAAGCATAGGTACCTTTTGTCTTTCTGGAAATGCTAGACCATGGAAATATGCTTCTGGTCTTAATTGCATTTTGAATCTGTAATTTTTGTGAGAGCTGCTCAGTTACTTTCAGGTTTTTGATCTCTCAAAAGAGGCTCCAGATAAGGTATTGCATCGTTTATATGCCAACCTT

[0020] The sequence of the amplification product (wild genotype) obtained by dgk2.2-F / R is as follows (SEQ ID NO.4): CCATGACATTTTCTCTCCTATGCATTATGTTGTAATCCTACAATATGGTATGAAGACCAAAGCTTCTTGCTTGAACTTGGGAAATCATTGTTCTGTTATATTTGTTCATCATTCCATCATACTGCTCCTTCATCTGTATTTCTTTTGGGCAGTAGATGGGTTTTTAACCTCTGCATTTGTACCTCTTGCAGCATTCGGTCCATCGTGTGTCTGAATTTGCCTAGTTTTTCTGGAGGTTTGAATCCTTGGGGCACACCTGGCACGAGGAGAGCAGAAGATGTGAGTTTCGATGTCTTTGTTTCTGTGAACTGAACATGCATAATTTGCAAGCCATGGAGATTAGTTGACTAATCTATTATGGCAATGTGCAGAGGGAGTTCACTGCACCTTTTGTCGACGATGGACTTCTTGAGGTTGTTGGCTTCCGTGACGCCTGGCACGGGCTGGTCCTGCTGGCCCCTAATGGACACGGCACTCGCATTGCCCAGGTATGAATTGCTTCTGCTACCTATTCCTCCTCCTCATGGAAATCCTTACATCTCAGAAAGGTGAACTCAGGCCCATCATGGTTTTGTGGTACACTAAAACGGGGTCAG

[0021] The sequence of the amplification product obtained by dgk2.2-F / R (homozygous mutant genotype) is as follows (SEQ ID NO.5): CCATGACATTTTCTCTCCTATGCATTATGTTGTAATCCTACAATATGGTATGAAGACCAAAGCTTCTTGCTTGAACTTGGGAAATCATTGTTCTGTTATATTTGTTCATCATTCCATCATACTGCTCCTTCATCTGTATTTCTTTTGGGCAGTAGATGGGTTTTTAACCTCTGCATTTGTACCTCTTGCAGCATTCGGTCCATCGTGTGTCTGAATTTGCCTAGTTTTTCTGGAGGTTTGAATCCTTAGGGCACACCTGGCACGAGGAGAGCAGAAGATGTGAGTTTCGATGTCTTTGTTTCTGTGAACTGAACATGCATAATTTGCAAGCCATGGAGATTAGTTGACTAATCTATTATGGCAATGTGCAGAGGGAGTTCACTGCACCTTTTGTCGACGATGGACTTCTTGAGGTTGTTGGCTTCCGTGACGCCTGGCACGGGCTGGTCCTGCTGGCCCCTAATGGACACGGCACTCGCATTGCCCAGGTATGAATTGCTTCTGCTACCTATTCCTCCTCCTCATGGAAATCCTTACATCTCAGAAAGGTGAACTCAGGCCCATCATGGTTTTGTGGTACACTAAAACGGGGTCAG

[0022] The present invention also provides a method for screening maize materials with dwarf traits, comprising the following steps: detecting molecular markers in the material to be tested, and screening out the materials containing the above molecular markers.

[0023] The present invention also provides a method for cultivating maize with dwarf traits, comprising the following steps: introducing the above molecular markers into the plants of the variety to be improved through field breeding or molecular breeding techniques to obtain homozygous dwarf plants.

[0024] The present invention isolated the diacylglycerol kinase ZmDGK2 gene from maize, and obtained two single-base mutations occurring in the 1st exon and the 10th exon of the gene ZmDGK2 by using a maize EMS mutant library, resulting in premature termination of ZmDGK2 translation and loss of gene function. Multi-point field trial investigations showed that the premature termination mutant materials zmdgk2.1 and zmdgk2.2 of ZmDGK2 had significantly reduced plant heights compared to the wild-type B73 inbred line, and thus two molecular markers were developed. The molecular markers developed based on the functional sites of the DGK gene in the present invention can accurately distinguish wild-type from dwarf mutants, and the detection results are stable and reliable, overcoming the problem that traditional phenotypic identification is easily affected by the environment. Through PCR amplification and sequencing analysis, dwarf materials can be quickly screened at the seedling stage, shortening the breeding cycle and significantly improving the breeding efficiency. The present invention provides an efficient and low-cost molecular tool for the genetic improvement of maize plant height, and has important practical value for modern maize breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is the Sanger sequencing information of the premature termination mutation site of the diacylglycerol kinase ZmDGK2 gene; (A) Mutation site C / T of the premature termination mutant zmdgk2.1. (B) Mutation site G / A of the premature termination mutant zmdgk2.2.

[0026] Figure 2 is the effect of the mutation of the diacylglycerol kinase ZmDGK2 gene on maize plant height; (A) Gene structure of ZmDGK2 and site information of the premature termination mutants zmdgk2.1 and zmdgk2.2. (B-C) Compared with the wild-type B73 inbred line, the premature termination mutant materials zmdgk2.1 and zmdgk2.2 showed significantly reduced plant heights in the fields of Sanya and Inner Mongolia. (D) Field plant height phenotypes of the wild-type B73, the premature termination mutant materials zmdgk2.1 and zmdgk2.2. DETAILED DESCRIPTION OF THE INVENTION

[0027] The following examples further illustrate the present invention, rather than limiting the present invention.

[0028] Example 1

[0029] In the present invention, an EMS mutant library with the maize inbred line B73 as the genetic background ( http: / / maizeems.qlnu.edu.cn / ) Screening and obtaining mutant materials of the diacylglycerol kinase gene ZmDGK2 (GRMZM2G094452); according to the gene mapping information of the EMS mutant materials, the mutants zmdgk2.1 (mutant number: EMS3-019ddc) and zmdgk2.2 (mutant number: EMS3-0aa6ff) have SNP variations in the 1st exon and the 10th exon of the gene ZmDGK2 (GRMZM2G094452), respectively.

[0030] Using the genomic DNA of the maize mutant materials zmdgk2.1 and zmdgk2.2 to be tested as templates respectively, PCR amplification of the 1st exon and the 10th exon of ZmDGK2 was carried out using the designed primer pairs. The PCR products were subjected to Sanger sequencing, and then the Sequencher 4.8 software was used to interpret the peak map for SNPs ( Figure 1 ).

[0031] Primer pair 1:

[0032] dgk2.1-F: CGCTTTTAGGATGGTGTGGAT (SEQ ID NO.6);

[0033] dgk2.1-R: AAGGTTGGCATATAAACGATGC (SEQ ID NO.7).

[0034] Primer pair 2:

[0035] dgk2.2-F: CCATGACATTTTCTCTCCTATGC (SEQ ID NO.8);

[0036] dgk2.2-R: CTGACCCCGTTTTAGTGTACCA (SEQ ID NO.9).

[0037] The PCR amplification system was as follows: 1 μL of each forward and reverse primer (10 μmol / L), 1 μL of genomic DNA, 0.5 μL of Phanta DNA polymerase, 12.5 μL of 2×Phanta Max buffer, 0.5 μL of dNTP mix, and made up to 25 μL with ddH2O. The PCR reaction program was: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 40 s, for a total of 35 cycles; over-extension at 72°C for 10 min; storage at 4°C. The PCR amplification reagents were purchased from Nanjing Novoprotein Biological.

[0038] Using the ZmDGK2 gene sequence on the maize inbred line B73 genome as a reference, primer pairs 1 (dgk2.1F / R) and 2 (dgk2.2 F / R) were designed with Primer5.0. Through Sanger sequencing, it was identified that the 220th variation site in the first exon of the mutant material zmdgk2.1 was C / T, resulting in the codon CAG mutating to the stop codon TAG; the 57th variation site in the tenth exon of the mutant material zmdgk2.2 was G / A, resulting in the codon TGG mutating to the stop codon TAG( Figure 2 in A). This indicates that this marker can well distinguish wild-type and mutant genes.

[0039] Example 2

[0040] By conducting randomized block trials in the fields of Sanya and Inner Mongolia, the plant heights of the wild-type material B73 and the homozygous mutants zmdgk2.1 and zmdgk2.2 were measured. Statistical analysis found that compared with the wild-type material B73, the plant heights of the homozygous mutants zmdgk2.1 (mutant number: EMS3-019ddc) and zmdgk2.2 (mutant number: EMS3-0aa6ff) were significantly reduced( Figure 2 in B-C). The field plant height phenotypes of wild-type B73, the premature termination mutant materials zmdgk2.1 and zmdgk2.2 are shown in Figure 2 D.

[0041] After verification, primer pairs dgk2.1-F / R and dgk2.2-F / R can clearly identify the homozygous SNP variations of the gene ZmDGK2 in the first exon and the tenth exon, and can be used as molecular markers for the assisted breeding of new maize dwarf varieties.

[0042] Therefore, the present invention discovers that the diacylglycerol kinase gene ZmDGK2 regulates maize plant height, and two pairs of molecular markers are developed for the genetic improvement of maize dwarf materials and the assisted breeding of new varieties.

[0043] SEQ ID NO.1(>ZmDGK2, GRMZM2G094452, B73_refV3)

[0044] AATGTAAGAAATACACACAAAAAAAAAGTCAAATCTTAAAATGGCGAACATGCGAGCCCCTAGAA

[0045] ACGATGACAACCAAGAGAGTGCCTCCTGTCCTCCTAACATCGATTTTCCTCCGGAATCGTAACCTCA

[0046] GAAAGCCAAATCTCATTCTCCGGCGCTCAGGCCCACCCAAGCGTCGACTCTCCCGATCCAATCGGT

[0047] ACCTCCTAGTTCCTCGCCGGCGGTAGCGGCGTCGGGCGAACAGCGGTGACTTGGCGAGGGCGCTT

[0048] GGCCGGCGACAATCATCCACTAGGTACGCTGTTCTTTCTCTTGCCTTCCTGCTGCGCGCAAACGAGC

[0049] GCCCCCAAGTTATATATGCATTCGGATCTGACCCAATTACGGTATATACATGTGTGGTATCCACTAAAT

[0050] TCGATTTTTGTTACCGGGTGTACATGTCTTTTACTCTGCCCGCTTCTGACTCTTTTCCTTCTCCATTTC

[0051] CTGTAATCTGATGTGCTGAAAAAATTATTCACGGGTATTTTATGTTGATGATGCTAAAATAGCAGATC

[0052] CATCATCTATTTGTGGGAGTATTTGATTTGCAAAACTCAAACTTTTCAAGTTTTGACCAACAATTGGT

[0053] CACATTATTATTCATGCTTCGTGCACAAGTGTTGCATCAAAAGATTCATATCCAACTTGACCAACAAT

[0054] TAGTGGAATTATTCATGCTTCATGCACAAATGTTGCACCAATAGGTTCATATCCAAAGCGCTTTTAGG

[0055] ATGGTGTGGATTCTGTTGTAAAGATAACATAATTCAGGACCACCCAATCAAAATATGCCTTGTATTCA

[0056] TGAATGGAGGGAGCAAACATTTGTAGCACCAGTGAGTTATCCCATTAAAACTATGAGAACGATTTTT

[0057] CAGGATGGAAGGTGAGGCAGAGACCGTGGTTGGTTCTTGTTCTAAACCATGTGGGCCTCTGGAGGA

[0058] CTACTACATTCCAGATTACATTCTGAAGCCAGGTGCCCAACAAGTACTTGTTGATCATGCGGCACCC

[0059] TGCCCCGTTGTAGTGTTCATCAACTCAAGATCTGGAGGCCAACTTGGAAGTAGTTTAATCAAAACAT

[0060] ATCGTGAGCTTCTCAATGAAGCA CAG GTACCTTTTGTCTTTCTGGAAATGCTAGACCATGGAAATAT

[0061] GCTTCTGGTCTTAATTGCATTTTGAATCTGTAATTTTTGTGAGAGCTGCTCAGTTACTTTCAGGTTTTT

[0062] GATCTCTCAAAAGAGGCTCCAGATAAGGTATTGCATCGTTTATATGCCAACCTTGAAAGGCTGAAGA

[0063] TGGAAGGAGACATTCTTGCAGTTCAAATTTGGAGGACACTGAGGCTAATTGTGAGTCATAACACTTT

[0064] CAGAATTTAGATTATTGTTATACTGTGGTATCTATCTCTTTTGCTTTCCCATTGTACTTATTTTGACTTA

[0065] ACAGATTTCAATTAGGTTGCAGGCGGTGATGGTACAGCTAGCTGGCTGCTTGGGGTAGTCAGTGAC

[0066] CTTAAGCTTTCCCACCCACCTCCAGTGGCAACTGTTCCTCTGGGAACCGGAAATAACCTCCCCTTTT

[0067] CATTTGGATGGGTAAGTGGCCAACATCATTCTTTTAAACAGATTTCCTTTATATTATTGTGTCGCTGAT

[0068] GTTTCACACAAAAGGAAAGTTGAAGTCATTTTTATGTTTTCTCATCCATGATTAAAGATAGAATAATC

[0069] AAATTCTCCTCTATCTCATCCATGGTGAAAAATTGAACATACTGATTAGTATACTGACGCTCCAATCA

[0070] CTATCATCCATCTACTTGTTACCTGTATACATATTGTCTAATACTTTGTACTCCTTTTGTTCCAAAAAAA

[0071] AACTTGGTCCTAAGTCAAACTATCTAATGTTTGACCAAATTTATATACTGGAATACTAGTGTTTATATT

[0072] ACCAACTAGTCGGTTGCCCGTGCATTGCGACGGCTTACAACAATATCCACGTAAACTATCCATCAAA

[0073] AAAATTCAAGATTTTTTATTGATTGTCTCCGCTCTCTGTATAATATATTTTTTGATTTGACTAACTGATG

[0074] TTATTGTTTACTCCATGCAAATATGTCTTGGTACAACACGACCAATGAAGTGAGCGATTAGAAGAGA

[0075] GTTCACAACGATTGACTGAATGAACAGAGATTATAAAATAACATAATTCCATCATACAAAGACCAAA

[0076] TAAGAGAAAGTTTGTGAGATCAAGTTTCTAAAATAAGTCCAATGAAGTCAAACTTATAAAAAAAGA

[0077] TGATCAAAATATGAAGTGATTGCTAAAGTCAGACATAAAAAAAAACTGAATGCGCTCCATATAAATT

[0078] ATGCTACTTCGTAGCAATTACTAACGTTTAAAACCAACAATAACCTTTCATTTTGCTGTTAGTGTGAC

[0079] AAATCATTGTTGCTCCATCCAATTCAGCAACTCAAACAACATGTAGTACATTGCGCCAATATCGTCTC

[0080] AAAAACAACCTAATGCTTGCAAGAGACAAGAACGTCGTGCCGTGCTTGGGCTGTAGCCTCGGCCC

[0081] GTAGTGCTGGCCCAGCCTGACACGATTATTTTTTTATTTTACAAAAAAACGTATATACATATATACAAT

[0082] TTATATTCAATATTAAAAACATCATGGCGTGATGTTCTACTCGTTAGACAGTTTCACCCAGTGTCTCC

[0083] CACACTTCTTCTATCAGGGAATGGGTTCGAACCCCACCTCCTGCACCGTTTTTTAACATTTTACGCTG

[0084] ATTTAATTAAATGGATCGATGGGCTAACGGGCTGGCCCGACACAGTTAGCAAGCCGGCATGACGTGT

[0085] TTGTACCATAGTTGTGGCCCGCGTGCATCTAGCCCATGTCGGGCGTCGTTACGCTGATTTAATTAAAT

[0086] GGATCGACGGGCTAATGGGCTGGCCCGATACAGTTAGCAGGCCGGCATGACGTGCCTGTGCCATAG

[0087] TTGTGGCCCGCGTGCATCTAGCCCATGTCGGGCGTCGTTTGGCATCTATAGACGTGCAGCGGATTAA

[0088] TTTGAAATAGCTGTGAGGGGTTATTTGTAAAAAAAATGACGCATGACGACCGTTGAAACTGGTGCT

[0089] TTAAGTATAGTATAGAATAAGTATCATTGGACACATCATGAAACATATTTTCATAATATAGTTATGTGTT

[0090] GTCATAAACATTAGCTATTTTTTGATTTTTCTATAGATTTGGTCAAACTTAAGATAACTTGATTTAGGA

[0091] CAAAACTAAAACATTATGTTTGTTTTGGAACAGAGGGACTACTTTGCATATGTTCAAGCTGTTGTTTT

[0092] AATATTTATTTTTATTGATAAGCTGCGGTGAGTGTAGAAATTTTGAAAAGTTGATGCACACTTTTGAG

[0093] TTCTATGATTCTATCAGGTCATATGCAAATCCAATTTAGTTGATAAAAAAACTATTTTCTTACGTGACT

[0094] TTAGCTTGCAATCTTGCATGCAATTGCTTCTACATGTAAGAAAAGATTTGTTTATGTTGAGTTCAACA

[0095] TTGCAGGGAAAGAAGAATCCTTCTACTGACCAAGAGGCTGTAAAATCATTCCTCGGGCTAGTAAAG

[0096] CATGCAAAAGAAATTAAGATTGATAGGTACTGCCCAAGAAAAAGAGCTCGTTGTTAGTTGTTTTAGG

[0097] TTATGTAATGATTTCCCTAAAGATGAAAATATTGCTAAATTTGTGTAATATTCTTCAGTTTGTAGCTTT

[0098] GTGCTTGTCTAGCCCATCCTTGTGGTAGCCTGTACGGAATTATTTTCTTCATAAAGAAAATGTCTCCT

[0099] GTGTGTTTGCGGCAAAAAAATTGTCTAATATTGCCTCCAATAATGGAGCATGTTCCTATGCTCATTTT

[0100] TCTGCTGCTTGTAGGATGCAAATTTATTTATTACAACAACAAAGCCTTTTAGTCTCAAGCAAGTTGTG

[0101] ATAGGTTAGAGTTGAAACCTAGTAGAAGTCACTAGTGAAGGTTGAGGCATGTGTATAGCTCTTTTCC

[0102] ATGCACTCATATTCACGGCTAAAACTTTGGGTATATTCCATCCTTTTAAGTCTCCTTTTACTATCTTTTT

[0103] CCATGTCAACTTCAGCCTTCTCTTGCTTCTCTTTCCCATGCAAAGTTATTTATTTACATTAAAAATAAT

[0104] TGAATAGTTAACCCTTCATCATCATGTCTTTCCTCAGTTTAAAGCATGCTTATTCTTTTCTCTCATGCT

[0105] AAGGCAGACACAATTTACTGTAAATTTTGATGTTTCAGTTGGCACATCATTTTGAGAATGCGAGTTC

[0106] CAGAGGAAGGTCCATGTGATCCTATTGCTCCATTAGATTTGCCTCATTCATTGCATGCGTTCCATCGT

[0107] GTCTCAAGTAGTGATTCTCTCAATATGGTAGTACACTCTTTCTATTGCAACTTTTTTTTGCTATGAATT

[0108] ATGTTTGCTTGAGAAAATGACATGCACCATTACCATCAAAGGCATTTGGTTAATGGATTTAATTCAAA

[0109] GCTTCCATGATCATTCTTGATTACTTCAGGAAGGTTACCACACATTCCGTGGAGGATTTTGGAATTAC

[0110] TTTAGTATGGGTAAGGCTTCAATGTTGTTCTCATCTTTATTGTTACACAAATGCGGCAAGACCATTAT

[0111] GTACCTATACTATCCAGTTGTTTATTTATATTTTTTATCTTGATATGTGATCCTGGGTAGTAGTTCATGCT

[0112] ACAAAGGCTAGTGTTTCGCCAACCCACTAAGGAAAGTAGTGCAAGTCTAGTGAGATTATTGTTATGG

[0113] CATCATTCTCTTTTTAATTAAATCTATATCTGGTGAGTTCTTAAGCAGATAACAAATTAAAGAATTCAG

[0114] GCACTAGAGTCAAACATGATGATCATGACTTGACATGTTGCAGGGATGGATGCAGAAGTGTCTTATG

[0115] CATTTCATTCTGAAAGGAAGAAGAATCCAGAGAAGTTCAAGAATCAGCTGACAAATCAGGTAAATG

[0116] TGTCATATTTTAGTTTTGTTACTATATTTTCAATCTGCTTGTTAAATAAATGACCACCTAGTTATGCTAA

[0117] AACTTTTGATTTGTTCATCTGCATCGATTATATAGGGTACATACGCTAAGCTTGGACTTAAACAAGGA

[0118] TGGTTTTGTGCTTCTCTTAGTCAGCCATCATCAAGGTAAAATTCGCAAAATTAGTCATAATCCTAAAT

[0119] CACAGACACTGCTCCCATGGTGCTTGTCTTTTGTGATATTTACTGTCTATTGTCTAGGAGACTAGGGT

[0120] CTAGGACTAATGGTGTATTGTATCTTGATCACATTTAATATACATTTGACTTCAGGCTGCAGGAGTGT

[0121] GGTAAGAGGAGTGCCATATTTGTCATTTTGTCACTCTCTAACTGTAGTGACTAATTCATTTTGTGTTTT

[0122] TTATTGCTCCAAATCACCAGTTCACCACCATCCTTTGCATTCTGAATTTACAATTTGGGAGACCAGCT

[0123] TTTTTTGGAGTATTATTTACAATACTAGATATTCTAATTCTGTCACTCTCGTGTATCTTCCAGGTTTCCC

[0124] ATTTCTCCTTTTGTGTTTAAAGGATAAACTAGATTGCTTTGTTTCAGGAACCTTGCTCAAATTGCAAA

[0125] AGTAAAGATCATGAAAAGAGCTGGTAGCCTCTGGGAGGAACTTCACATTCATCACAGGTAAACGTA

[0126] GGCGATTTTTTTCTGTTATGGGTCCTAGTTCAGAGGGGAATAACCAACGCCGGAAAGATAGCCGGA

[0127] CAGAGTCGGCTAGGGGGAGACTAGAGTAGAGAGAGGTGTTAGATTAAACTTCTTTGATTGATTCCC

[0128] CCTTAAGGAGGTACAACTCATCCTTATATAGATAGGAAGACTTGGCCCCCAAGTAACTAACTCAATC

[0129] TTATCTTTTAGAACGAAACCAGCGCACATTTGCGCCCAGGGAAGCCCTCCACGTCATCATGATGCAT

[0130] ATCATCCGTAGGATCTTGATCGAACAGTAAGACCTTTCTCGAAGATTGTCCTCAGTCCTCTGCTATAT

[0131] AAAAAGAAGAAAATGAGTGGAGTCTCACATCGTACCAGTCTGCCAAGCAAACAGTCTGCCAGGCA

[0132] AAATACCAAAGACCAAAGGAGACTCGCGGAAAGAAAAGAGAGAGTAAAAAAGATAAAAAGAAGA

[0133] AAGAGGGAGTCACAGTTGTTCAACATACATCTCTGAACATCCTCTGCTATATAAAAAGAAGAAAATG

[0134] AGTGGGGTCTCACATCGTACCAGTCTGCCAAGAAAAATATTAAAGACCAAAAAGACCCCACGGAA

[0135] AGAAGAGAGATAGCAAAAAAGAGAGAAAAAAGGAACACAAAGAGTCACGGTTGTTCAACATACA

[0136] AAGCAAAGATATCCTTGGTGATCCAGGAGTGCAGCAGGGGGAAAGAGGTACGAACCGAACCTTAA

[0137] CAAAACACCATCTTGGTTCCATAGCCTTGCCATTTGCTTCTCGAGAGGCATCTCCATATTCCCTTCGC

[0138] CAGCTTAAGAAAAATATAAAGTTCCAACAATTATGCAATCTTTTCCTTCTTCAAGTCCTAATATTGTA

[0139] CTGACTGCAAGGTGTAGGAAAAAGGTTACCTGAACTCTCTCCATGCATAAATAAAAACATGCCGTAT

[0140] GAATAACTTACATGGAAAAATATAATTGTACCATCAAAACATCTTTCGAAGGTACAAAATAGCAGTC

[0141] CTAATGACCTACTCTCTACAGTCAGTCAGTCACAGATAAGTGACCTTTTGCAATTGTTTTAGGCCAA

[0142] CCATTTCAAACATAGGCTGCATATTACCTTTTATTCAGAGTAGCAAATACACTTTCATGAAGTAAAAT

[0143] ATGATGGATGGAAGAATCCATTTTTCCTTTAATATTAGGTATATATATATCCGGGAAAATGGTATGAGTA

[0144] TTTTTGTTTCTCAATGTAGTTCCATAAAAATATATTTTTGCTCACTGTCTCCAATCACCAAAAAAATTT

[0145] AGGGCAAGATTTAGTACCATCCACTTGATCACTATAAATATGTACCTCATCCAACCCACTGACAATAT

[0146] AATACATACATACATAACAGAAAATTAACTACCCAAGCCCATAGCTAACCAATGGCCCTTTTGAGAG

[0147] CGCGCCAACGGCGCACCATGCTTTCTAGTCTAATCTAATCCCAAGTAACTAACTCAATCTTATCTAAT

[0148] TTAATCCCAAGTAACTAACTCAATCTTATCTAATCTAATCTTTATCCACTAATCCTAACCCTGAGCCTA

[0149] TCCCTATGGCTTCCATGCTCTGGGCTACCGCCCCATGACATTCCTCCACCCTTTGAAGACAGCTTGTC

[0150] CTTGAGCTGTAATGGGGTGCATGGCAACAGCCTTGCTGTCTATCTTGATCTGCACGTGAAAGTGCAA

[0151] CCCTAACTAGAAGCCTTTTATATCTCGACTTTAATTTATTTGCAAAATAAAAATAAAAGAACCTAACA

[0152] TATGTCAAAGGTTGGGTCCCCCTGGATCCCATGGGAAACAACCTCCTCCCAAGCTGCAAAGGCCCA

[0153] AGACGTGGCAGCTCACCAGTCCTATGGCTGGAGCTGTCGCGAGTTTCCCAGTTGAAGGACAATCCC

[0154] ACCCAACCCAGACCATGTTCTTGGTTGGTCCACATGGGTATAAGATCAGGACGGTGAATTGGGGGT

[0155] GGAGGAATCGGCCAACCCTGCTGAAGGCCTTGCCTCCCCAAGTGTCGCGTGTTTTCAACGCGCAAT

[0156] ATCGATGCAAGTCGTTGTGCCAGGGTCGCCGCTTGACGTGCTTGATGCTGCGCCAGGACTGATCGG

[0157] CCACCAGACTCCACCTTTGCCGCGAGTAGATCACGAAGTTGGAGGATAGTAGATCGGGCTTGCTCC

[0158] AAATCTATCCTGGCACGAGTGAGCTCCTCACGCCAGGTCTGTCATCATCCATGGCAGAAGTCATGGG

[0159] AAGACTCACTATACCAGATGTTATGGGTCCTAGTTCATAGGGCGATAACCAGCACTGGGAAGATAGT

[0160] CGGGCGGAGTCGGCTTTGGGGGAAACTAGAGCAGAGAGGTATTAGATTAAACTTGATTGATTCCCC

[0161] TTTAAGGAGGTACATCTCATCCTTGTAACTAACTCAAGGTATAGATCGGAAGACTTGCCCCCCAAGT

[0162] AATTAACTCAATCTTATCTAATCTAATCCTAAGTAATTAACTCAATCTTATCTAATCCAATCCCAAGTA

[0163] ACTAACTCAATCTTATCTAATCTAATCACAATTAACTAACTCAATTTTATCTAATCTAATATTTATCCAC

[0164] TAATCCTAACCCTGAGCCTATCTCTATGGCCTCCATACTCTGGGCTGCCACCCCATGACATTTTCTCTC

[0165] CTATGCATTATGTTGTAATCCTACAATATGGTATGAAGACCAAAGCTTCTTGCTTGAACTTGGGAAAT

[0166] CATTGTTCTGTTATATTTGTTCATCATTCCATCATACTGCTCCTTCATCTGTATTTCTTTTGGGCAGTAG

[0167] ATGGGTTTTTAACCTCTGCATTTGTACCTCTTGCAGCATTCGGTCCATCGTGTGTCTGAATTTGCCTA

[0168] GTTTTTCTGGAGGTTTGAATCCT TGG GGCACACCTGGCACGAGGAGAGCAGAAGATGTGAGTTTC

[0169] GATGTCTTTGTTTCTGTGAACTGAACATGCATAATTTGCAAGCCATGGAGATTAGTTGACTAATCTAT

[0170] TATGGCAATGTGCAGAGGGAGTTCACTGCACCTTTTGTCGACGATGGACTTCTTGAGGTTGTTGGCT

[0171] TCCGTGACGCCTGGCACGGGCTGGTCCTGCTGGCCCCTAATGGACACGGCACTCGCATTGCCCAGG

[0172] TATGAATTGCTTCTGCTACCTATTCCTCCTCCTCATGGAAATCCTTACATCTCAGAAAGGTGAACTCA

[0173] GGCCCATCATGGTTTTGTGGTACACTAAAACGGGGTCAGTCTTCCCATCTAAATCTGATTTCACCATC

[0174] CTGTTTCACAGGCCCATAGGATCCGCTTTGAATTCCACAAAGGCGCAGCAGAGCACACGTTCATGA

[0175] GGGTTGACGGAGAGCCCTGGAAGCAGCCCCTTCCCAAGGACGATGACACGGTTGTGGTGGAAATC

[0176] TCTCACCTTGGGCAGGTCACCATGCTAGCAAATGAGCCGTGCAGGTCCAAGAGCGTCAACGACCA

[0177] GTCGTCACTGTCGCCGGCGCACGACAGTCATGGTGATTACAATGATATCGCTGAGGAGGACGAAGA

[0178] CGAGTGGGAGGACGGCAGGAGGAAGTTTGGGGCGGCAGATACATTTAAGATTCCTGACGAGATTG

[0179] ACATTGCTCATTTCAGTTGAGCTTTGACTAGTGCATCGTCGTGTGCGTTCTGATCATACGAGTATGGT

[0180] TTGGTTTATAACCATCTCCTTTCTTTTGATTTCATATATTTGCGATATGATTGTTGCAGCCATGTGTAAG

[0181] TGAATCCTAGGTAACCAAGTGCTGCGTTGCTATATATGACCAATGTATGTATAGATTCAGTGGAACGG

[0182] TAAAAACACGCGAACTCTACGGTTTTCTCCTTTTGTTGTTACCAGTTCAATTGAACAATATAGCGGG

[0183] TGTTTGGTTTGGGTAATCACGCTATTCAAAATATTCAAAATAAGGTGATACATTATGGGTCTATTATGT

[0184] AATCACTCCATTCTAAATAAGTTGGTGCATCATAGGTCTATTCTTTAAATTTGGTGGGATGACTCTATT

[0185] CTTTATATCTAGTTAATCATTGATCAACTCATTATATTTCACAAACTAAACAAAAAGTCAGCAGTAAG

[0186] AAGACGATGGACTAATTTATTTCTCAAACTAAACATCCAACAAATTTAGTGGGATGACCTCATTTCTA

[0187] ATATTATTACTAATTAACTATGAGGAATAAGGTGTTGATGGGTCAACTTATTTTATTCTATAAACCAAA

[0188] CAAAAAATGAGGAGTGATGTAAACCCTAAATATGACATTAATGAAGTTAGCAGTGCTTTTATTGTGT

[0189] AATACCCAATTTGTAAGAAAATATTAAAGGAGAAATTATTTCCTTTATATATATATATGTTATCTCTAATT

[0190] ACTATCACATGTGAACATCTCATTTAAAAACAAATAGTTAATAAAGGGACATGCCACTAAATTGTGCA

[0191] TCATGATGGAGTTTTTGCTTGTTTGTGCACTTGATAATAAAATAATAACAATAGAAATATAATAATGAG

[0192] GTGGAAAAATTTGAATTTAAACCAAAGGTGCACTTTAGGGAATTGAGAACAGTATAAAGAAAATAG

[0193] GGAATAAATATTATGCAACACAAGATAAACATTTTTGGCATGTCCAAATTGTACTCTTAATCAAAGAA

[0194] TAATTTACCACAGTGTTGAATTTGAAATTTGAATTCAAAATGTGAAGGGAAAACAAATCAGAAAAAT

[0195] AAAAAAGAAAATAAAATAAAAA

[0196] The above are only the preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be regarded as limitations on the present invention. The protection scope of the present invention should be defined by the scope of the claims. For those of ordinary skill in the art, without departing from the spirit and scope of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as within the protection scope of the present invention.

Claims

1. A molecular marker associated with dwarfing of maize plants, characterized in that: The molecular marker is a mutation of C at position 1099 to T in the sequence shown in SEQ ID NO.1, or a mutation of G at position 8405 to A in the sequence shown in SEQ ID NO.

1.

2. A specific primer for detecting the molecular marker according to claim 1, characterized in that: The specific primers include a primer pair dgk2.1-F / R for detecting the 1099th mutation site: dgk2.1-F:CGCTTTTAGGATGGTGTGGAT; dgk2.1-R:AAGGTTGGCATATAAACGATGC; and primer pair dgk2.2-F / R for detecting mutation site 8405; dgk2.2-F:CCATGACATTTTCTCTCCTATGC; dgk2.2-R:CTGAACCCCGTTTTAGTGTACCA.

3. Use of the molecular marker described in claim 1 or the specific primer described in claim 2 in identifying or assisting in identifying the plant height trait of corn.

4. Use of the molecular marker described in claim 1 or the specific primer described in claim 2 in the genetic improvement of dwarf maize materials or the auxiliary breeding of new varieties.

5. A method for identifying or assisting in identifying a corn plant height trait, characterized in that: The method comprises the following steps: using the genomic DNA of the corn to be tested as a template, performing PCR amplification on dgk2.1-F / R or dgk2.2-F / R using the primers described in claim 2 to obtain an amplified product, sequencing the amplified product, and identifying the corn plant height according to the base type of the mutation site.

6. The method according to claim 5, characterized in that If the type of the base at the 369bp position of the amplified product obtained using the primer pair dgk2.1-F / R is C, the corn material to be tested is a wild genotype; if it is T, the plant height of the corn material to be tested is shortened; if the type of the base at the 248bp position of the amplified product obtained using the primer pair dgk2.2-F / R is G, the corn material to be tested is a wild genotype; if it is A, the plant height of the corn material to be tested is shortened.

7. A method for screening corn materials with dwarf stalk traits, characterized in that: The method comprises the following steps: detecting the molecular markers in the material to be tested, and screening out the material containing the molecular markers as claimed in claim 1.

8. A method for cultivating corn with a dwarf stalk trait, characterized in that: The method comprises the following steps: introducing the molecular markers described in claim 1 into plants of the variety to be improved through field breeding or molecular breeding technology to obtain homozygous dwarf plants.

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