Maize deep sowing-resistant molecular marker and application

Abstract

The invention discloses a corn deep sowing-resistant molecular marker and application thereof. The nucleotide sequence of the corn deep sowing-resistant molecular marker ZmCRK10Type A is shown as SEQ ID NO: 4. The gene ZmCRK10 encodes cysteine-rich receptor kinase and participates in regulation and control of plant growth and development, and overexpression of a T04 transcript can increase the mesocotyl length under corn deep sowing treatment and improve the deep sowing resistance of corn. Resequencing of associated populations subsequently finds that when large-fragment structural variation exists in a corn ZmCRK10 promoter region and a transposon with the length of about 121.7 kb exists, expression of a T04 transcript of the ZmCRK10 can be remarkably improved, corn mesocotyl elongation is promoted, and the ZmCRK10Type A can be used as a corn deep sowing-resistant molecular marker.

Description

[0001] The application is a divisional application of Chinese Invention Patent Application No. 2024113783516, “Corn Deep Sowing Tolerance Gene ZmCRK1, Molecular Marker and Application”. ZmCRK10 and Application”. TECHNICAL FIELD

[0002] The present application relates to the field of plant genetic engineering, and in particular to a corn deep sowing tolerance molecular marker and application. BACKGROUND

[0003] About two-thirds of the corn planting area in China is distributed in arid and semi-arid regions. The natural precipitation in these regions is low, the soil surface water evaporates quickly, and the soil water content is low, which seriously affects early spring corn planting, and causes corn to be extremely susceptible to drought stress at the germination and emergence stages, ultimately affecting yield. Proper deep sowing is beneficial for the seed to fully utilize deep soil water, and ensures the germination and emergence of corn in drought planting environment, which is one of the important ways for corn seedling drought resistance. However, so far, the cloning of corn deep sowing tolerance QTL has not been reported. Therefore, it is of great significance to fully utilize corn germplasm resources, analyze the mechanism of corn deep sowing tolerance, screen and breed new corn varieties with deep sowing tolerance, for corn drought resistance and seedling protection, stable yield and yield increase in arid and semi-arid regions.

[0004] Under the condition of proper deep sowing, the emergence rate of corn is significantly positively correlated with the elongation of mesocotyl and hypocotyl. Corn varieties with longer mesocotyl or hypocotyl can ensure faster seed emergence under deep sowing, and the elongation of mesocotyl plays a major role, which is greater than that of hypocotyl. The mesocotyl of corn refers to the part between the root and hypocotyl of corn seedling. The elongation of mesocotyl is affected by multiple internal and external factors such as light, sowing depth, hormones, lignin metabolism and variety difference. Cysteine-rich receptor-like kinase (CRK) is a large class of plant receptor-like kinases. CRK is reported to regulate ROS generation, calcium ion signal, MAPK cascade, ABA response and callose deposition, and is widely involved in plant growth and development, immune response and abiotic stress. Some studies have found that Arabidopsis CRK mutants affect seed germination, emergence, rosette leaf size and root development, indicating that CRK-mediated callose deposition may be involved in the regulation of cell turgor-driven elongation. In summary, the corn CRK gene may play a role in regulating the elongation and development of mesocotyl, and has potential application value in the breeding of new corn varieties with deep sowing tolerance. CRK CRK In addition to molecular biological breeding, it is also of great significance to use genetic methods to breed and improve corn varieties with deep sowing tolerance. Screening of corn deep sowing tolerance related gene molecular markers can effectively improve the efficiency of breeding corn varieties with deep sowing tolerance.

[0005] In addition to molecular biological breeding, it is also of great significance to use genetic methods to breed and improve corn varieties with deep sowing tolerance. Screening of corn deep sowing tolerance related gene molecular markers can effectively improve the efficiency of breeding corn varieties with deep sowing tolerance. SUMMARY​

[0006] The present application aims at overcoming the deficiencies of the prior art, and provides a corn and deep sowing tolerance molecular marker and application, and a corn deep sowing tolerance gene ZmCRK10 The encoded cysteine-rich receptor kinase is involved in regulating plant growth and development. ZmCRK10 There are two main transcripts, overexpression ZmCRK10 of T04 the transcript can promote the elongation of the mesocotyl under the deep sowing condition of corn. The corn deep sowing tolerance molecular marker ZmCRK10 Type A includes a transposon (containing the structure of multiple LTR long terminal repeat transposons) with a length of about 121.7 kb identified by the marker. The presence of the marker significantly promotes the development of the mesocotyl of corn, and the mechanism is to promote the expression of the downstream ZmCRK10-T04 transcript and promote the elongation of the mesocotyl.

[0007] To achieve the above-mentioned purpose, the technical scheme designed by the present application is as follows: The present application provides a corn deep sowing tolerance gene ZmCRK10 The nucleotide sequence of the deep sowing tolerance gene ZmCRK10 is shown as SEQ ID NO: 1.

[0008] The present application also provides a deep sowing tolerance transcript ZmCRK10 of the corn deep sowing tolerance gene ZmCRK10-T04 The CDS nucleotide sequence of the transcript ZmCRK10-T04 is shown as SEQ ID NO: 3.

[0009] The present application also provides a deep sowing tolerance protein ZmCRK10-T04 encoded by the deep sowing tolerance transcript ZmCRK10-T04 The amino acid sequence of the protein is shown as SEQ ID NO: 2.

[0010] The present application also provides a primer pair for obtaining the deep sowing tolerance transcript ZmCRK10-T04 The primer pair is as follows: ZmCRK10-T04-F0: 5'-GTCGACAAACGCACTAGTATCCCGGGA TGGATTTCGACCCTAAGCTC-3', ZmCRK10-T04-R0: 5'-GGGTACATTTGGCGCGCCTTCCCGGG TCTAGGATACAGTTCACTGAGTGTA-3'.

[0011] The present application also provides a deep sowing tolerance protein ZmCRK10-T04 encoded by the deep sowing tolerance transcript ZmCRK10-T04Or the application of the deep-sowing tolerance protein ZmCRK10-T04 in improving the deep-sowing tolerance of corn.

[0012] The application also provides a deep-sowing tolerance transcript ZmCRK10-T04 Or the application of the deep-sowing tolerance protein ZmCRK10-T04 in breeding new deep-sowing tolerance corn varieties.

[0013] The application also provides a deep-sowing tolerance molecular marker ZmCRK10 Type A The nucleotide sequence of the deep-sowing tolerance molecular marker ZmCRK10 Type A is shown in SEQ ID NO: 4.

[0014] The application also provides a deep-sowing tolerance molecular marker ZmCRK10 Type A in improving the deep-sowing tolerance of corn and assisting in breeding.

[0015] The application also provides a primer pair for obtaining the deep-sowing tolerance molecular marker ZmCRK10 Type A , and the primer pair is respectively: ZmCRK10 Type A -F1: 5'-ACCTCATCTCCTTCATGTGTGGTC-3', ZmCRK10 Type A -R1: 5'-GCATAGGTGGAATCGTCGCAG-3'.

[0016] The application also provides the application of a kit in identifying corn varieties with excellent deep-sowing tolerance, and the kit comprises the primer pair described above.

[0017] The application also provides a method for identifying corn varieties with excellent deep-sowing tolerance by using the kit described above, comprising the following steps: (1) Extracting the DNA of the corn variety to be detected; (2) Designing the primer pair as follows: ZmCRK10 Type A -F1: 5'-ACCTCATCTCCTTCATGTGTGGTC-3', as shown in SEQ ID NO: 5 ZmCRK10 Type A -R1: 5'-GCATAGGTGGAATCGTCGCAG-3', as shown in SEQ ID NO: 6; (3) Performing PCR amplification by using the DNA as the template; (4) Electrophoresis: the target band of 403 bp appears in the amplification, which indicates that the variety has strong deep-sowing tolerance.

[0018] The principle of this invention: This invention utilizes corn conversion technology to obtain corn. ZmCRK10-T04 Overexpression material. The overexpression material, isolated negative control material, and transformed background material KN5585 were seeded at a depth of 15 cm. The mesocotyl length of the overexpression material was found to be significantly higher than that of the wild type. Further gel section analysis of the mesocotyls revealed that the mesocotyl cell length of the overexpression material was significantly higher than that of the isolated negative control and background materials, indicating... ZmCRK10-T04 It may positively regulate mesocotyl elongation in maize under deep-sowing conditions by promoting cell elongation. Associated population resequencing revealed this in maize. ZmCRK10 Large structural variations exist in the promoter region, including a 121.7 kb (ZmCRK10) segment. Type A ) and 63.3 kb (ZmCRK10 Type B The transposable of ZmCRK10 Type A The presence of significantly increased ZmCRK10 of T04 The expression of transcripts promotes the development of the maize mesocotyl and can serve as a molecular marker for maize tolerance to deep planting.

[0019] The beneficial effects of this invention are: This invention utilizes the mesocotyl length and 550,000 SNP markers from maize populations treated with a 15 cm deep planting depth to perform GWAS analysis, identifying a gene associated with deep planting tolerance. ZmCRK10 This gene encodes a cysteine-rich receptor kinase involved in regulating plant growth and development. Overexpression of the transcript... ZmCRK10-T04 It can promote mesocotyl elongation under deep planting conditions and improve maize's tolerance to deep planting. The presence of structural variation in the 121.7 kb transposon in its promoter region significantly promotes maize mesocotyl development, and as a molecular marker for maize deep planting tolerance, it can effectively improve the efficiency of breeding deep-planting tolerant maize varieties. Attached Figure Description

[0020] Figure 1 A schematic diagram illustrating the construction and identification of maize overexpression materials; In the figure, 'a' represents the overexpression vector pZZ0153-UBIp- ZmCRK10-T04 -3HA construction diagram Figure b shows the identification results of DNA insertion into the overexpression vector of the transgenic material. c represents genetically modified materials. ZmCRK10 Expression level identification plot (the expression level in KN5585 is set to 1).

[0021] Figure 2 for ZmCRK10-T04 Phenotypic analysis of overexpression material and control at a seeding depth of 15 cm; In the figure, a represents the mesocotyl phenotype images of the overexpressing material and the control under a 15 cm deep seeding treatment, where Bar = 1 cm. b is a statistical analysis of the mesocotyl length of the overexpressing material and the control under 15 cm deep seeding treatment. c shows mesocotyl sections of the overexpression material and control after seeding at a depth of 15 cm. The sections were stained with Calcofluor; blue indicates the cell wall location. Bar = 50 μm. d is a schematic diagram of the mesodermal cell length of the overexpression material and the control under 15 cm deep seeding treatment. Each point represents the average length of 10 mesodermal cells.

[0022] Figure 3 for ZmCRK10 Allelic variation analysis diagram; In the figure, 'a' represents the associated group. ZmCRK10 Promoter structural variations and ZmCRK10 Type A Genotyping gel electrophoresis b is ZmCRK10 Type A Association analysis diagram between genotype and hypocotyl length in associated populations. c is ZmCRK10 Type A Genotype and ZmCRK10-T04 Association analysis diagram of transcript abundance.

[0023] Figure 4 For F 2:3 ZmCRK10 in the population, BC3F3 population and RIL F8 population Type A A diagram illustrating how natural variations affect maize's tolerance to deep sowing; a) with a sowing depth of 15 cm, using LIAO5263×CML423 as the parent F 2:3 A schematic diagram of the mesocotyl length in the population. b is a seeding depth of 15 cm, with LIAO5114×CIMBL83 as the parent F. 2:3 A schematic diagram of the mesocotyl length in the population. c is a schematic diagram of the mesocotyl length of the BC3F3 population with LY042×CML423 as the parent at a sowing depth of 15 cm. A schematic diagram of the mesocotyl length of the RIL F8 population with BY815×KUI3 as the parent at a sowing depth of 15 cm.

[0024] Figure 5 This diagram illustrates the use of primers and kits to detect the genotype of different maize varieties and the length of the mesocotyl in deeply sown maize. a is ZmCRK10 Type A Gel images for identifying maize varieties by genotype. b is ZmCRK10Type B Gel images for identifying maize varieties by genotype. c is ZmCRK10 Type A Mesocot length of maize varieties with different genotypes at a sowing depth of 15 cm. d is ZmCRK10 Type B Mesocotyl length of maize varieties with different genotypes at a sowing depth of 15 cm. Detailed Implementation

[0025] The present invention will now be described in further detail with reference to specific embodiments, so that those skilled in the art can understand it.

[0026] Example 1: Maize deep-planting tolerance gene ZmCRK10 Positioning Genome-wide association analysis (GWAS) was performed using genotypic data from 380 natural maize populations (data source: http: / / maizego.org / Resources.html) combined with mesocotyl length data from maize under a 15 cm seeding depth. Target genes were located using the maize genome annotation file. ZmCRK10 The expression of this gene in the mesocotyl increases with increasing sowing depth, and its nucleotide sequence is shown in SEQ ID No: 1; the amino acid sequence of the cysteine-rich receptor kinase encoded by this gene is shown in SEQ ID No: 2.

[0027] Example 2: Corn ZmCRK10-T04 Construction of overexpression vectors Based on the expression vector pZZ0153-UBIp-3HA, the deep-seated transcript was expressed through homologous recombination. ZmCRK10- T04 The cDNA fragment is inserted downstream of the UBI promoter and between the 3HA tag, and expressed by the UBI promoter. Figure 1 Electrophoresis detection and sequencing analysis showed that maize was successfully obtained. ZmCRK10-T04 The overexpression vector, named pZZ0153-UBIp- ZmCRK10- T04 -3HA, the specific experimental steps are as follows: 1. Deep-seated transcripts ZmCRK10-T04 Amplification: according to ZmCRK10-T04 A pair of homologous recombination primers was designed based on the cDNA sequence for PCR amplification. The primer sequences are as follows: ZmCRK10-T04-F0: 5'-GTCGACAAACGCACTAGTATCCCGGGA TGGATTTCGACCCTAAGCTC-3', ZmCRK10-T04-R0: 5'-GGGTACATTTGGCGCGCCTTCCCGGGTC TAGGATACAGTTCACTGAGTGTA-3'; Gene amplification was performed using Vazyme Phanta Max Super-Fidelity DNA polymerase to obtain PCR products, and sequencing was performed to obtain deep-sowing transcript ZmCRK10-T04 The CDS nucleotide sequence of which is shown in SEQ ID NO: 3; the reaction system is as follows (total volume 20 μL): The reaction procedure is as follows: 95°C for 3 min; 95°C for 15 s; 58°C for 30 s; 72°C for 60 s; 72°C for 5 min; 35 cycles. 2. Linearized vector preparation: restriction endonuclease XmaI Single enzyme digestion of pZZ0153-UBIp-3HA vector to linearize the circular vector; 3. Detection and recovery of PCR product and vector digestion product: agarose gel electrophoresis was used to detect the PCR product and vector digestion product. The Omega Bio-tek Gel Extraction Kit was used to recover the target fragment; 4. Gel recovery product recombination: homologous recombination was performed using the Vazyme ClonExpress II One Step Cloning Kit; the reaction system is as follows: The reaction solution was collected at the bottom of the centrifuge tube after brief centrifugation by gently pipetting. It was placed in a 37°C water bath for 30 min and then left to stand on ice until room temperature. 5. Recombination product transformation: the recombination product was transformed into E. coli DH5α competent cells, and the molecular cloning experiment guide was referred to.

[0028] 6. Sequencing identification: after colony PCR and plasmid PCR, the positive plasmid was selected for sequencing to be correct, indicating that the maize ZmCRK10 overexpression vector pZZ0153-UBIp- ZmCRK10-T04 -3HA was successfully obtained. Primer F: 5'-CACATACTCAGACAGGCAGAGTTG-3', Primer R: 5'-TCTGGAACGTCGTATGGG-3'.

[0029] Example 3 Transgenic maize line obtained The corn plants obtained in Example 2 were used as the transformation background, and the corn plants were transformed by the overexpression vector pZZ0153-UBIp-3HA obtained from Jiangsu KeyGene Biotech Co., Ltd. ZmCRK10 The overexpression vector pZZ0153-UBIp- ZmCRK10-T04 -3HA obtained from Jiangsu KeyGene Biotech Co., Ltd. The T0 generation of transgenic seeds obtained from the company were cultivated to obtain the T1 generation, and the T1 positive plants were identified by DNA detection of the overexpression vector insertion and the seeds were harvested, and the T2 generation was cultivated. PCR detection was continued on the T2 generation plants to determine that the target gene did not occur genetic segregation loss, and finally two stably inherited positive homozygous transgenic corn families and corresponding negative segregation materials were obtained; the PCR detection primers were the same as in Example 2.6. ZmCRK10-T04 Stable genetic positive homozygous transgenic corn families and corresponding negative segregation materials were obtained; the PCR detection primers were the same as in Example 2.6.

[0030] Example 4 ZmCRK10-T04 Expression amount detection of transgenic corn plants The transgenic corn plants and wild type were planted respectively ZmCRK10-T04 The overexpression corn materials and wild type were planted respectively, and the three-week-old corn leaves were taken, and the RNA was extracted by the Trizol method. After the extracted RNA was digested by DNase I, reverse transcription was performed using Promega MLV reverse transcriptase, and RT-PCR detection was performed using the extracted cDNA, and the corn Actin gene was used as a control.

[0031] The detection results showed that ZmCRK10-T04 the gene was successfully introduced into corn Figure 1 c).

[0032] The primers used in the detection process are as follows: qZmCRK10-F: 5'-GGGACCTGAAGGCAAATAAC-3', qZmCRK10-R: 5'-GTCCCAACAACTCTGCCTGT-3'; qZmActin-F: 5'-GCTGGATCTTGCTGGCCGTG-3', qZmActin-R: 5'-AGGCGCCACGACCTTGATCT-3'; Example 5 ZmCRK10-T04 Detection of deep sowing tolerance phenotype of transgenic corn At present, the 15 cm sowing depth treatment can better reflect the deep sowing tolerance of different varieties. And 27 core population materials were randomly selected for different sowing depth treatment (10, 15 and 18 cm), and the results showed that under the 15 cm sowing depth treatment, the hypocotyl length had the highest correlation with the emergence rate, and the linear relationship fitting effect was the best (R 2 =0.62), which could better reflect the seedling emergence ability under deep sowing.

[0033] Therefore, ZmCRK10-T04 Positive transgenic material, negative isolate, and background material KN5585 were simultaneously planted in pots (length × width × height = 50 cm × 35 cm × 25 cm, with drainage holes at the bottom) with a 5 cm layer of soil at the bottom, and covered with a 15 cm layer of soil. Water was drawn from the bottom of the pot. After culturing the materials in a dark room at 28°C for 10 days, the seedlings were carefully removed, the soil near the mesocotyl was removed to preserve the integrity of the mesocotyl as much as possible, and the mesocotyl length of the seedlings was measured by photographing. The results showed that overexpression... ZmCRK10-T04 Significantly promotes the elongation of the maize mesocotyl under deep sowing treatment ( Figure 2 a, b). Mesocotyl samples were taken 1 cm below the coleoptile and gel-sectioned to a thickness of 60 µm. The mesocotyl sections were stained with Calcofluor, and cell wall signals were observed under 405 nm excitation light. Figure 2 c) It was found that the length of mesocotyl cells in the overexpression material was significantly higher than that in the isolated negative control and background material. Figure 2 d).

[0034] In summary, ZmCRK10-T04 It may positively regulate the elongation of the maize mesocotyl under deep sowing conditions by promoting cell elongation. ZmCRK10-T04 Genetically modified corn has a stronger ability to tolerate deep planting.

[0035] Example 6: Molecular marker ZmCRK10 for maize tolerating deep planting Type A Filtering and association analysis 1. ZmCRK10 in the associated population Type A Genotyping and association analysis with mesocotyl length For maize materials B73 and SK whose sequencing sequences have been published ZmCRK10 Sequence analysis revealed a transposon approximately 121.7 kb long in the B73 promoter region, and another transposon approximately 63.3 kb long with significant sequence differences in the SK promoter region. Primers were designed based on the sequences to identify transposon insertions in the ZmCRK10 promoter region of the associated population. Figure 3 a). Used to identify 121.7 kb (ZmCRK10). Type A The primer pair for the transposon is ZmCRK10. Type A -F1 / R1 is as follows (amplified partial sequence): ZmCRK10 Type A -F1:5'-ACCTCATCTCCTTCATGTGTGGTC-3', ZmCRK10 Type A -R1: 5'-GCATAGGTGGAATCGTCGCAG-3'.

[0036] PCR amplification system as follows (total volume 20 μL) : Reaction procedure: 95°C 3 min; 95°C 30 s; 58°C 30 s; 72°C 45 s; 72°C 5 min; 35 cycles.

[0037] 2. ZmCRK10 Type A Genotype and association population in the hypocotyl length and ZmCRK10-T04 Transcript abundance association analysis ZmCRK10 Type A -F1 / R1 by PCR to identify ZmCRK10 Type A genotype, distinguished from another genotype.

[0038] The results found that ZmCRK10 Type A genotype material under the deep planting treatment of the hypocotyl length is significantly higher than another genotype ( Figure 3 b). Promoter region structural variation often has a significant impact on the expression of the gene. By analyzing the existing association population leaf transcriptome data, it is found that the Type A genotype material, ZmCRK10 T04 transcript abundance is significantly higher than another genotype ( Figure 3 c). It is shown that ZmCRK10 Type A genotype may promote the elongation of the maize hypocotyl by increasing the expression of ZmCRK10 T04 transcript; therefore, the nucleotide sequence of the corn deep planting tolerance molecular marker ZmCRK10 Type A is shown as SEQ ID NO: 4.

[0039] Example 7 F 2:3 population, BC3F3 population and NIL population ZmCRK10 Type A Natural variation affects corn deep planting tolerance Using ZmCRK10 Type A genotype molecular marker ( Figure 3 a) and hypocotyl length phenotype, select the parent to construct F 2:3 population, BC3F3 population and NIL population F8 separation population, 15 cm deep planting phenotype analysis (same as example 5), verify the variation site.

[0040] The results show that under the condition of 15 cm deep planting, ZmCRK10 Type A genotype homozygous single plant has a longer hypocotyl ( Figure 4 ​​), indicating ZmCRK10 Type A Allelic variation is a superior allelic variation that promotes mesocotyl elongation and is suitable for deep-planting tolerance. It can be used as a molecular marker for deep-planting tolerance in maize to improve the efficiency of breeding deep-planting tolerant maize varieties.

[0041] Example 8: Identification of maize varieties with different mesocotyl lengths using kit primers 1. Reagent kit and instructions for identifying maize varieties with excellent deep-planting tolerance The kit includes the molecular marker ZmCRK10 for obtaining maize deep-planting tolerance. Type A Primer pair ZmCRK10 Type A -F1 / R1, where primer pair ZmCRK10 Type A -F1 / R1 is: ZmCRK10 Type A -F1:5'-ACCTCATCTCCTTCATGTGTGGTC-3', ZmCRK10 Type A -R1:5'-GCATAGGTGGAATCGTCGCAG-3' The method for identifying maize varieties with excellent deep-planting tolerance using the above-mentioned kit includes the following steps: (1) Extract DNA from the maize variety to be tested; (2) The primer pairs are designed as follows: ZmCRK10 Type A -F1:5'-ACCTCATCTCCTTCATGTGTGGTC-3', ZmCRK10 Type A -R1:5'-GCATAGGTGGAATCGTCGCAG-3' (3) Perform PCR amplification using the above DNA as a template; (4) Electrophoresis: The amplification showed a target band of 403 bp, indicating that the variety has a strong ability to withstand deep sowing.

[0042] 2. Identification of maize varieties with different mesocotyl lengths using a kit. Verify ZmCRK10 Type A Allelic variations can be used as molecular markers for maize deep-planting tolerance. Then, ZmCRK10 was selected using a kit. Type A and ZmCRK10 Type B Different genotypes of maize varieties were treated with a 15 cm deep sowing depth. Figure 5 (a, b) includes commonly used maize inbred lines Chang7-2, B73, BY804, and 18-599. Results show that ZmCRK10 Type AThe genotype corn variety under the deep sowing treatment is significantly higher than ZmCRK10 Type B Genotype corn variety ( Figure 5 c, d). Provide an effective kit product for the preliminary identification of deep sowing corn varieties.

[0043] Other parts not described in detail are prior art. Although the above examples make a detailed description of the present application, it is only a part of the embodiments of the present application, not all embodiments, and other embodiments can be obtained without creativity on the basis of the present embodiments, which are within the scope of the present application.

Claims

1. A molecular marker for deep sowing tolerance in maize, ZmCRK10 Type A characterized in that The corn deep sowing tolerance molecular marker ZmCRK10 Type A The nucleotide sequence of ZmCRK10 is shown as SEQ ID NO:

4.

2. A corn deep planting tolerance molecular marker ZmCRK10 of claim 1 Type A Use in improving corn deep planting tolerance and in assisted breeding.

3. A primer pair for obtaining the deep sowing tolerance molecular marker ZmCRK10 of claim 1, characterized in that: the forward primer is SEQ ID NO: 1; and the reverse primer is SEQ ID NO:

2. Type A The primer pair is: ​ ZmCRK10 Type A - F1 : 5'-ACCTCATCTCCTTCATGTGTGGTC-3', ZmCRK10 Type A - R1 : 5'-GCATAGGTGGAATCGTCGCAG-3'.

4. Use of a kit in identifying corn varieties with superior ability to withstand deep planting, characterized in that: The kit comprises the primer pair of claim 3.

5. A method for identifying corn varieties with superior ability to withstand deep planting using the kit of claim 4, characterized by: The method comprises the following steps: (1) extracting DNA of the corn variety to be detected; (2) designing the primer pair as follows: ZmCRK10 TypeA - F1 : 5'-ACCTCATCTCCTTCATGTGTGGTC-3', ZmCRK10 Type A - R1 : 5'-GCATAGGTGGAATCGTCGCAG-3'; (3) performing PCR amplification with the above DNA as a template; (4) electrophoresis: the target band of 403 bp appears, indicating that the variety has strong deep sowing resistance.

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