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CN-122012781-A - Primer combination of molecular marker for regulating ZmDFOT gene of corn day flowering time and application thereof

CN122012781ACN 122012781 ACN122012781 ACN 122012781ACN-122012781-A

Abstract

The invention belongs to the technical field of genetic engineering, and particularly relates to a primer combination of a molecular marker of ZmDFOT gene for regulating and controlling corn day flowering time and application thereof. The primer combination consists of a PCR primer pair 1 and a PCR primer pair 2, wherein the nucleotide sequence of the PCR primer pair 1 is shown as SEQ ID NO. 3-SEQ ID NO.4, and the nucleotide sequence of the PCR primer pair 2 is shown as SEQ ID NO. 5-6. The nucleotide sequence of ZmDFOT gene is shown as SEQ ID NO. 1. The excellent haplotype of the corn daily flowering time regulating gene ZmDFOT can be used for identifying and screening materials with early corn daily flowering time, and is suitable for cultivating new corn varieties with early daily flowering time and high temperature avoidance.

Inventors

  • TANG JIHUA
  • CHEN XIAOYANG
  • ZHANG SHOUFENG
  • DING DONG
  • CAO XUDONG
  • YANG HAO

Assignees

  • 河南农业大学

Dates

Publication Date
20260512
Application Date
20260211

Claims (8)

  1. 1. The primer combination of the molecular marker of ZmDFOT gene for regulating the corn flowering time per day is characterized by comprising a PCR primer pair 1 and a PCR primer pair 2; The nucleotide sequence of the PCR primer pair 1 is shown as SEQ ID NO. 3-SEQ ID NO. 4; The nucleotide sequence of the PCR primer pair 2 is shown as SEQ ID NO. 5-SEQ ID NO. 6; the nucleotide sequence of ZmDFOT gene is shown as SEQ ID NO. 1.
  2. 2. The primer combination according to claim 1, wherein the PCR primer pair 1 is a primer pair for amplifying SNP1 and SNP2, and the PCR primer pair 2 is a primer pair for amplifying SNP 3; The SNP1 is positioned at the 5066 th position of the ZmDFOT gene, and the base of the position is G or A; The SNP2 is positioned at the 4531 st position of the ZmDFOT1 gene, and the base of the position is C or T; The SNP3 is positioned at 2478 of the ZmDFOT gene, and the base at the 2478 is G or A.
  3. 3. A kit for identifying the time to day flowering characteristics of corn, comprising the primer combination of any one of claims 1-2.
  4. 4. Use of the primer combination of any one of claims 1-2 or the kit of claim 3 for identifying the time-to-day flowering of maize.
  5. 5. The use according to claim 4, wherein the step of identifying the time-to-day flowering profile of maize is as follows: Extracting genome DNA of corn materials to be detected; performing PCR amplification on the genomic DNA by using the PCR primer pair 1 and the PCR primer pair 2; analyzing the PCR amplification product sequence, judging genotypes of positions 5066, 4531 and 2478 of ZmDFOT gene in the corn material to be detected, and further identifying the corn day flowering time character of the corn to be detected.
  6. 6. The use according to claim 5, wherein if genotypes at 5066, 4531 and 2478 of the ZmDFOT gene in the test maize material are GG, CC, GG, the maize daily flowering-time trait of the test maize material is identified as early as the daily flowering-time, and if genotypes at 5066, 4531 and 2478 of the ZmDFOT gene in the test maize material are AA, TT, AA, the maize daily flowering-time trait of the test maize material is identified as late as the daily flowering-time; The early flowering time of the day means that the time of the corn reaching the powder scattering peak period is 7:00-8:00 per day; The late flowering time refers to the time when the corn reaches the peak of powder scattering, which is 8:30-9:30 per day.
  7. 7. The use according to claim 6, wherein corn material with genotypes GG, CC, GG having early day flowering time is selected by simultaneously detecting genotypes at 5066, 4531 and 2478 of ZmDFOT gene in corn material to be tested.
  8. 8. The use according to claim 7, wherein said corn material with early daily flowering time is used for breeding high temperature-resistant, high yield corn hybrids.

Description

Primer combination of molecular marker for regulating ZmDFOT gene of corn day flowering time and application thereof Technical Field The invention belongs to the technical fields of molecular biology and genetic engineering, and particularly relates to a primer combination of a molecular marker of ZmDFOT gene for regulating and controlling corn day flowering time and application thereof. Background With global climate change, high temperature has become one of the important abiotic stress factors affecting corn yield. It is predicted that each 1 ℃ increase in average air temperature will result in a 7.4% reduction in corn yield. The impact of high temperature on corn is mainly focused on the ear differentiation period and the loose powder spinning period. In the yellow-Huaihai region, the summer corn grows in the same season in rainy and hot period, the scattered pollen silk period is often overlapped with high-temperature weather, so that the corn tassel glume flowers are prevented from being opened, the anther cracking rate and the pollen leakage rate are reduced, the pollen vitality is reduced, fertilization cannot be carried out, and the fruiting rate and the yield are reduced. Solves the influence of high temperature in the flowering phase on corn production, and is an important requirement for guaranteeing corn yield and national grain safety. At present, the high-temperature heat resistance of corn is mainly improved by utilizing high-temperature resistant gene resources. However, extremely high temperature weather is increasingly frequent, and the ventilation and air permeability of the field corn population are poor, and the strategy of improving the high temperature heat resistance of corn through a high temperature resistant way is difficult to completely solve the adverse effect of high temperature on corn production. Day flowering time (Diurnal Flower Opening Time, DFOT) refers to the time of caryopsis opening in a day. By advancing the glume flower opening time in one day of corn, the high temperature can be avoided, the glume flowers are promoted to open at a lower temperature, anther cracking and pollen scattering are completed, and the pollen vigor and female ear setting rate are further improved. The utilization of early flowering germplasm resources to avoid high temperature is an effective measure for solving the problem of high temperature injury of corns. To realize efficient and accurate screening of early flowering traits, development of specific molecular marker primer combination products aiming at regulating key genes of the traits is needed to realize efficient and accurate molecular marker assisted selection of the traits. Disclosure of Invention In order to develop a specific molecular marker primer combination product aiming at a key gene for regulating and controlling the corn daily flowering time property, the invention develops a specific molecular marker primer combination for the key gene for regulating and controlling the corn daily flowering time property, and realizes efficient and accurate screening of the property. The invention develops a molecular marker for identifying the haplotype of the day-flowering time gene and a primer thereof, thereby identifying germplasm resources with early day-flowering time. The invention provides a primer combination of a molecular marker of ZmDFOT gene for regulating and controlling corn day flowering time and application thereof. To achieve the above object, the above technical problems are solved. The invention adopts the following technical scheme. In order to identify key genes of day flowering time and further develop germplasm resources with early day flowering time so as to solve the problem of maize flowering-phase high temperature injury, the invention provides a primer combination of a molecular marker for regulating ZmDFOT gene of the day flowering time of maize, which consists of a PCR primer pair 1 and a PCR primer pair 2. The nucleotide sequence of the PCR primer pair 1 is shown as SEQ ID NO.3 (SNP 1-2F:5 '-CCAAGCACCTTCTCACAAGT-3') and SEQ ID NO.4 (SNP 1-2R:5 '-GACGCATCATTTGTGTAGCG-3'). The nucleotide sequence of the PCR primer pair 2 is shown as SEQ ID NO.5 (SNP 3F: 5'-TGATCAACCGATGGGCATAC-3') and SEQ ID NO.6 (SNP 3R: 5'-AGATATCACCTGGCAGAGCA-3'). The nucleotide sequence of ZmDFOT gene is shown as SEQ ID NO. 1. In order to develop a specific molecular marker primer combination product aiming at a key gene for regulating and controlling the corn daily flowering time property, the invention develops a specific molecular marker primer combination for the key gene for regulating and controlling the corn daily flowering time property, and realizes efficient and accurate screening of the property. The ZmDFOT gene for regulating and controlling the daily flowering time of the corn is cloned and functionally identified, so that a core molecular target is provided for solving the problem of high temperature injury in the flower