A molecular marker associated with maize plant dwarfing and use thereof

Abstract

The application discloses a molecular marker related to corn plant dwarfing and application thereof. The molecular marker is that a C at the 1099th position in the sequence shown in SEQ ID NO. 1 is mutated into T, or a G at the 8405th position in the sequence shown in SEQ ID NO. 1 is mutated into A. For the mutation, two pairs of molecular marker primers dgk2.1-F / R and dgk2.2-F / R are developed, and wild type and dwarf mutant can be accurately distinguished through PCR and sequencing. The molecular marker is helpful for assisted selection of new corn dwarf varieties.

Description

Technical Field

[0001] This invention belongs to the field of molecular genetic marker technology, specifically relating to a molecular marker related to dwarfing of maize plants and its application. Background Technology

[0002] Corn (Zea mays L.), as an important food, feed, and industrial raw material, is my country's largest crop and occupies a vital position in the national economy. Appropriate plant height can effectively reduce lodging and increase planting density, thus contributing to high corn yields.

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

[0004] Previous studies have shown that maize plant height is a complex quantitative trait controlled by a series of small-effect, multi-gene sets. Although there are many QTL loci regulating maize plant height, only a small number of genes have been successfully cloned. Therefore, there is an urgent need to identify key genes regulating maize plant height that can be applied to breeding. Summary of the Invention

[0005] The purpose of this invention is to provide a molecular marker related to maize plant dwarfing and its application, providing a new breeding approach with greater application potential for genetic improvement of maize plant height and dense-density breeding.

[0006] This invention is achieved through the following technical solution:

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

[0008] The present invention also provides a specific primer for detecting the above-mentioned molecular marker, said specific primer comprising 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:CTGAACCCCGTTTTAGTGTACCA.

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

[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 in the assisted breeding of new varieties.

[0016] The present invention also provides a method for identifying or assisting in the identification of maize plant height, comprising the following steps: using the genomic DNA of the maize to be tested as a template, performing PCR amplification using the above-mentioned primer pairs dgk2.1-F / R or dgk2.2-F / R to obtain amplification products, sequencing the amplification products, and identifying maize plant height based on the base type of the mutation site.

[0017] Specifically, the type of base at the 369bp position of the amplification product obtained using primer pair dgk2.1-F / R is C; if it is T, the maize material being tested is stunted. Similarly, the type of base at the 248bp position of the amplification product obtained using primer pair dgk2.2-F / R is G; if it is A, the maize material being tested is stunted.

[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 amplified product (homozygous mutant genotype) obtained by dgk2.2-F / R is as follows (SEQ ID NO.5): CCATGACATTTTCTCTCCTATGCATTATGTTGTAATCCTACAATATGGTATGAAGACCAAAGCTTCTTGCTTGAACTTGGGAAATCATTGTTCTGTTATATTTGTTCATCATTCCATCATACTGCTCCTTCATCTGTATTTCTTTTGGGCAGTAGATGGGTTTTTAACCTCTGCATTTGTACCTCTTGCAGCATTCGGTCCATCGTGTGTCTGAATTTGCCTAGTTTTTCTGGAGGTTTGAATCCTTAGGGCACACCTGGCACGAGGAGAGCAGAAGATGTGAGTTTCGATGTCT TTGTTTCTGTGAACTGAACATGCATAATTTGCAAGCCATGGAGATTAGTTGACTAATCTATTATGGCAATGTGCAGAGGGAGTTCACTGCACCTTTTGTCGACGATGGACTTCTTGAGGTTGTTGGCTTCCGTGACGCCTGGCACGGGCT GGTCCTGCTGGCCCCTAATGGACACGGCACTCGCATTGCCCAGGTATGAATTGCTTCTGCTACCTATTCCTCCTCCTCATGGAAATCCTTACATCTCAGAAAGGTGAACTCAGGCCCATCATGGTTTTGTGGTACACTAAAACGGGGTCAG

[0022] The present invention also provides a method for screening maize materials with dwarf stalk characteristics, comprising the following steps: detecting molecular markers in the test material and screening out materials containing the above-mentioned molecular markers.

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

[0024] This invention isolated the diglyceride kinase ZmDGK2 gene from maize. Using a maize EMS mutant library, two single-base mutations were obtained at exons 1 and 10 of the ZmDGK2 gene, causing premature termination of ZmDGK2 translation and loss of gene function. Multi-site field trials showed that the ZmDGK2 premature termination mutant materials zmdgk2.1 and zmdgk2.2 had significantly lower plant heights compared to the wild-type B73 inbred line, leading to the development of two molecular markers. These molecular markers, developed based on the functional sites of the DGK gene, can accurately distinguish between wild-type and dwarfing mutants, providing stable and reliable results and overcoming the susceptibility to environmental influences in traditional phenotypic identification. Through PCR amplification and sequencing analysis, dwarfing materials can be rapidly screened at the seedling stage, shortening the breeding cycle and significantly improving breeding efficiency. This invention provides an efficient and low-cost molecular tool for the genetic improvement of maize plant height, possessing significant practical value for modern maize breeding. Attached Figure Description

[0025] Figure 1 These are Sanger sequencing information for the premature termination mutation sites of the diglyceride kinase ZmDGK2 gene; (A) C / T mutation site in the premature termination mutant zmdgk2.1. (B) G / A mutation site in the premature termination mutant zmdgk2.2.

[0026] Figure 2 The effects of mutations in the diglyceride kinase ZmDGK2 gene on maize plant height are as follows: (A) ZmDGK2 gene structure and site information of the premature termination mutants zmdgk2.1 and zmdgk2.2. (BC) Compared with the wild-type B73 inbred line, the premature termination mutant materials zmdgk2.1 and zmdgk2.2 showed significantly reduced plant height in fields in Sanya and Inner Mongolia Autonomous Region. (D) Field plant height phenotypes of wild-type B73, premature termination mutant materials zmdgk2.1 and zmdgk2.2. Detailed Implementation

[0027] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0028] Example 1

[0029] This invention utilizes an EMS mutant library with maize inbred line B73 as the genetic background. http: / / maizeems.qlnu.edu.cn / Mutant materials of the diglyceride kinase gene ZmDGK2 (GRMZM2G094452) were obtained through screening. According to the gene localization information of the mutant materials by EMS, mutants zmdgk2.1 (mutant number: EMS3-019ddc) and zmdgk2.2 (mutant number: EMS3-0aa6ff) have SNP variations in the first exon and the tenth exon of the gene ZmDGK2 (GRMZM2G094452), respectively.

[0030] Using genomic DNA from the maize mutants zmdgk2.1 and zmdgk2.2 as templates, PCR amplification of exon 1 and exon 10 of ZmDGK2 was performed using designed primers. The PCR products were then subjected to Sanger sequencing, and SNPs were interpreted using Sequencher 4.8 software. 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:CTGAACCCCGTTTTAGTGTACCA (SEQ ID NO. 9).

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

[0038] Based on the ZmDGK2 gene sequence in the maize inbred line B73, primer pairs 1 (dgk2.1F / R) and 2 (dgk2.2F / R) were designed using Primer 5.0. Sanger sequencing identified a C / T mutation at position 220 of exon 1 in the mutant material zmdgk2.1, resulting in a CAG codon mutation to the stop codon TAG; similarly, a G / A mutation at position 57 of exon 10 in the mutant material zmdgk2.2 resulted in a TGG codon mutation to the stop codon TAG. Figure 2 The A in the figure indicates that the marker can effectively distinguish between wild-type and mutant genes.

[0039] Example 2

[0040] Randomized block experiments were conducted in Sanya and Inner Mongolia Autonomous Region. Plant height was measured for wild-type material B73 and homozygous mutants zmdgk2.1 and zmdgk2.2. Statistical analysis revealed that compared to wild-type material B73, the plant height of homozygous mutants zmdgk2.1 (mutant ID: EMS3-019ddc) and zmdgk2.2 (mutant ID: EMS3-0aa6ff) was significantly reduced. Figure 2 (BC in the text). Field plant height phenotypes of wild-type B73, premature termination mutant materials zmdgk2.1 and zmdgk2.2 are shown in the figure. Figure 2 D in the middle.

[0041] The primer pairs dgk2.1-F / R and dgk2.2-F / R have been verified to clearly identify homozygous SNP variations of the ZmDGK2 gene in exon 1 and exon 10, and can be used as molecular markers for the auxiliary breeding of new dwarf maize varieties.

[0042] Therefore, this invention discovered that the diglyceride kinase gene ZmDGK2 regulates maize plant height, and developed two pairs of molecular markers for genetic improvement of dwarf maize materials and 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] TTCTATGATTCTATCAGGTCATATGCAAATCCAATTTAGTTGATAAAAAAAACTATTTTCTTACGTGACT

[0094] TTAGCTTGCAATCTTGCATGCAATTGCTTCTACATGTAAGAAAAGATTTGTTTATGTTGAGTTCAACA

[0095] TTGCAGGGAAAGAAGAATCCTTCTACTGACCAAGAGGCTGTAAAATCATTCCTCGGGCTAGTAAAG

[0096] CATGCAAGAGAATTAAGATTGATAGGTACTGCCCAAGAAAGAGGCTCGTTGTTAGTTGTTTTAGG

[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] CATTTCATTCTGAAAGGAAGAAGAATCCAGAGAAGTTCAAGAATCAGCTGACAATTCAGGTAAATG

[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] CCTTAAGGAGGTACAACTCATCCTTATATAGAGAGACTTGGCCCCCAAGTAACTAACTCAATC

[0129] TTATCTTTTAGAACGAAACCAGCGCACATTTGCGCCCAGGGAAGCCCTCCACGTCATCATGATGCAT

[0130] ATCATCCGTAGGATCTTGATCGAACAGTAAGACCTTTCTCGAAGATTGTCCTCAGTCCTCTGCTATAT

[0131] AAAAAAGAAAATGAGTGGAGTCTCCACATCGTACCAGTCTGCCAAGCAAACAGTCTGCCAGGCA

[0132] AAATACCAAAGACCAAAGGAGACTCGCGGAAAGAAAGAGAGAGAAAAAAGATAAAAAAAGA

[0133] AAGAGGGAGTCACAGTTGTTCAACATACATCTCTGAACATCCTCTGCTATAAAAAGAAGAAAATG

[0134] AGTGGGGTCTCACATCGTACCAGTCTGCCAAGAAAAATATTAAAGACCAAAAGACCCCACGGAA

[0135] AGAGAGAGATAGCAAAAAGAGAGAAAAAGGAACACACAAAGAGTCACGGTTGTTCAACATACA

[0136] AAGCAAAGATATCCTTGGTGATCCAGGAGTGCAGCAGGGGAAAGAGGTACGAACCGAACCTTAA

[0137] CAAACACCATCTTGGTTCCATAGCCTTGCCATTTGCTTCTCGAGAGGCATCTCCATATTCCCTTCGC

[0138] CAGCTTAAGAAAAATATAAAGTTCCAACAATTATCAATCTTTTCCTTCTTCAAGTCCTAATATTGTA

[0139] CTGACTGCAAGGTGTAGGAAAAAGGTTACCTGAACTCTCTCCATGCATAAATAAAAACATGCCGTAT

[0140] GAATAACTTACATGGAAAAATATAATTGTACCATCAAAACATCTTTCGAAGGTACAAAATAGCAGTC

[0141] CTAATGACCTACTCTCTACAGTCAGTCAGTCACAGATAAGTGACCTTTTGCAATTGTTTTAGGCCAA

[0142] CCATTTCAAACATAGGCTGCATATTACCTTTATTCAGAGTAGCAAATACACTTTCATGAAGTAAAAT

[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] CGGGCGGAGTCGGCTTTGGGGGAAACTAGAGCAGAGAGGTTAGATTAAACTTGATTGATTCCCC

[0161] TTTAAGGAGGTACATCTCATCCTTGTAACTAACTCAAGGTATAGATCGGAAGACTTGCCCCCCAAGT

[0162] AATTAACTCAATCTTCTTAATCTATCCTAAGTAATTAACTCAATCTTCTTAATCAATCCCAAGTA

[0163] ACTAACTCAATCTTATCTAATCTACACAATTAACTAACTCAATTTTCTTAATTCTAATTTATCCACAC

[0164] TAATCCTAACCCTGAGCCTATCTCTATGGCCTCCATACTCTGGGCTGCCACCCCATGACATTTTCTCTC

[0165] CTATGCATTATGTTGTAATCCTACAATATGGTATGAAGACCAAAGCTTCTTGCTTGAACTTGGGAAAT

[0166] CATTGTTCTGTTATATTGTTCATCATTCCATCATACTGCTCCTTCATCTGTATTTCTTTTGGGCAGTAG

[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] TCATGATGGAGTTTTTGCTTGTTTGTGCACTTGATAATAAAAATAATAACAATAGAAATATAATAATGAG

[0192] GTGGAAAAATTTGAATTTAAACCAAAGGTGCACTTTAGGGAATTGAGAACAGTATAAAGAAAATAG

[0193] GGAATAAATATTATGCAACACAAGATAAACATTTTTGGCATGTCCAAATTGTACTCTTAATCAAAGAA

[0194] TAATTTACCACAGTGTTGAATTTGAAATTTGAATTCAAAATGTGAAGGGAAAACAAATCAGAAAAAT

[0195] AAAAAAGAAAATAAAATAAAAA

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

Claims

1. Use of a molecular marker associated with the dwarfing of corn plants in the identification or assisted identification of the height trait in corn plants, characterized in that, The molecular marker is that the base at position 369 of the sequence shown in SEQ ID NO. 2 is mutated from C to T, or the base at position 248 of the sequence shown in SEQ ID NO. 4 is mutated from G to A; if the base at position 369 of the sequence shown in SEQ ID NO. 2 is C, the corn material to be tested is wild genotype, if it is T, the plant height of the corn material to be tested is dwarf; if the base at position 248 of the sequence shown in SEQ ID NO. 4 is G, the corn material to be tested is wild genotype, if it is A, the plant height of the corn material to be tested is dwarf.

2. Use according to claim 1, characterized in that, The detection primer of the molecular marker related to the dwarf of the corn plant comprises a primer pair dgk2.1-F / R for detecting the mutation site at position 369 of the sequence shown in SEQ ID NO. 2: dgk2.1-F: CGCTTTTAGGATGGTGTGGAT; dgk2.1-R: AAGGTTGGCATATAAACGATGC; and a primer pair dgk2.2-F / R for detecting the mutation site at position 248 of the sequence shown in SEQ ID NO. 4: dgk2.2-F: CCATGACATTTTCTCTCCTATGC; dgk2.2-R: CTGACCCCGTTTTAGTGTACCA.

3. The use of a molecular marker related to the dwarfing of corn plants in the genetic improvement of corn dwarf materials or in the assisted selection of new varieties, characterized in that, The molecular marker is that the base at position 369 of the sequence shown in SEQ ID NO. 2 is mutated from C to T, or the base at position 248 of the sequence shown in SEQ ID NO. 4 is mutated from G to A; if the base at position 369 of the sequence shown in SEQ ID NO. 2 is C, the corn material to be tested is wild genotype, if it is T, the plant height of the corn material to be tested is dwarf; if the base at position 248 of the sequence shown in SEQ ID NO. 4 is G, the corn material to be tested is wild genotype, if it is A, the plant height of the corn material to be tested is dwarf.

4. A method of identifying or aiding in the identification of a maize 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, using the primer pair dgk2.1-F / R or dgk2.2-F / R in claim 2 to perform PCR amplification, obtaining an amplification product, sequencing the amplification product, and identifying the plant height of the corn according to the base type of the mutation site; using the base type at position 369 of the amplification product obtained by using the primer pair dgk2.1-F / R, if it is C, the corn material to be tested is wild genotype, if it is T, the plant height of the corn material to be tested is dwarf; using the base type at position 248 of the amplification product obtained by using the primer pair dgk2.2-F / R, if it is G, the corn material to be tested is wild genotype, if it is A, the plant height of the corn material to be tested is dwarf.

5. A method of screening maize material for a dwarf trait, comprising, The method comprises the following steps: detecting a molecular marker related to corn plant dwarfing in a material to be tested, wherein the molecular marker is that a base at position 369 of a sequence shown in SEQ ID NO. 2 is mutated from C to T or a base at position 248 of a sequence shown in SEQ ID NO. 4 is mutated from G to A; if the base at position 369 of the sequence shown in SEQ ID NO. 2 is C, the corn material to be tested is a wild genotype, if it is T, the corn material to be tested is dwarfed in plant height; if the base at position 248 of the sequence shown in SEQ ID NO. 4 is G, the corn material to be tested is a wild genotype, if it is A, the corn material to be tested is dwarfed in plant height.

6. A method of growing corn having a dwarf trait, comprising, The method comprises the following steps: introducing a molecular marker related to corn plant dwarfing into a plant of a variety to be improved by field breeding or molecular breeding technology to obtain a homozygous dwarf plant, wherein the molecular marker is that a base at position 369 of a sequence shown in SEQ ID NO. 2 is mutated from C to T or a base at position 248 of a sequence shown in SEQ ID NO. 4 is mutated from G to A; if the base at position 369 of the sequence shown in SEQ ID NO. 2 is C, the corn material to be tested is a wild genotype, if it is T, the corn material to be tested is dwarfed in plant height; if the base at position 248 of the sequence shown in SEQ ID NO. 4 is G, the corn material to be tested is a wild genotype, if it is A, the corn material to be tested is dwarfed in plant height.

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