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CN-119307643-B - Molecular marker and application of novel functional allelic variant Sbdth1 of sorghum heading stage gene SbDTH1

CN119307643BCN 119307643 BCN119307643 BCN 119307643BCN-119307643-B

Abstract

The invention discloses a molecular marker of a novel functional allelic variant Sbdth of a sorghum heading stage gene SbDTH1 and application thereof. The invention provides a functional allelic variant in a gene SbDTH for regulating and controlling the heading stage of sorghum, and a transposon is inserted at the 4 th base after a stop codon TGA. Allelic variants may allow for an extended heading time of sorghum. Molecular markers for high throughput marker molecular marker detection have been developed based on allelic variants. The molecular marker has the advantages of flexibility, high efficiency, high flux, low cost and the like. The molecular marker can be used for detecting and screening allelic variants, is used for molecular marker breeding and breeding new sorghum varieties suitable for heading period, and has important significance for enlarging regional planting of sorghum and improving yield.

Inventors

  • WU YUYE
  • Zhang Xiurun
  • ZHOU MIN
  • Xia Qimiao
  • WANG HAILIAN
  • ZHANG HUAWEN
  • LIU SHUBING

Assignees

  • 山东农业大学

Dates

Publication Date
20260512
Application Date
20241108

Claims (6)

  1. 1. A method of detecting a novel functional allelic variant Sbdth of sorghum heading date gene SbDTH1, comprising the steps of: the amplification reaction was performed using the following primer combinations: Forward primer 1F having the sequence SEQ ID NO.1: 5'-GCTCACACTCAACCACATGC-3' and/or Forward primer 2F with the sequence of SEQ ID NO.2: 5'-GCTACAGGCACAGATGACGA-3'; Reverse primer 1R with the sequence of SEQ ID NO.3: 5'-TTTGTTGACCTCGGTCTGAA-3'; The allelic variant Sbdth is a transposon inserted in the sorghum chromosome 8 gene SbDTH1 at the 4 th base after the termination codon TGA, and the allelic variant Sbdth has a later heading; Amplifying by using the primers 2F and 1R, and judging that the sorghum contains Sbdth genotype 1 if 675bp band can be amplified; And/or, using the primers 1F and 1R for amplification, if a 615bp band can be amplified, determining that the sorghum contains a wild type SbDTH.sup.1 genotype.
  2. 2. The method of claim 1, wherein the transposon has a base number of 6755bp.
  3. 3. The method of claim 1, wherein the transposon is inserted at a site 2314bp from the start codon of the gene SbDTH 1.
  4. 4. Use of a method according to any one of claims 1-3, for detecting whether a sorghum variety, strain or germplasm resource is Sbdth genotype 1.
  5. 5. Use of a method according to any one of claims 1-3 for detecting the distribution of said allelic variants in sorghum germplasm resources.
  6. 6. Use of a method according to any one of claims 1-3, characterized in that molecular markers for the heading stage of sorghum assist in selective breeding.

Description

Molecular marker and application of novel functional allelic variant Sbdth1 of sorghum heading stage gene SbDTH1 Technical Field The invention relates to the technical field of crop molecular marker assisted breeding, in particular to a molecular marker of a novel functional allelic variant Sbdth1 of a sorghum heading stage gene SbDTH and application thereof. Background Sorghum (Sorghumbicolor) is the fifth most popular cereal crop in the world, a staple food for more than 5 million people in africa and asia. In addition to being used as a precious food source, sorghum plays an important role in the brewing industry, the feed industry, the pigment industry, biomass energy sources and the like, so that the demand of China on sorghum is increased year by year. Therefore, it is extremely important to further increase the yield of sorghum and enlarge the planting area of sorghum. The heading (flowering) time is one of the important agronomic traits of crops and successful control of flowering time is critical to crop production because it ensures that the plant is propagated under optimal conditions. Cereal sorghum is generally selected to bloom early to avoid drought or low temperatures, ensuring reproduction, while other sorghum types, such as sweet sorghum, forage sorghum and energy sorghum, are selected for a longer vegetative growth period to achieve higher biomass yields. The diversity of sorghum flowering time under long sunlight conditions shows that multiple genes can be selected to cultivate photoperiod insensitive varieties. Thus, flowering time is a major limiting factor in crop breeding in terms of improving yield and regional adaptability. Sorghum is a typical short-day plant, the difference of photoperiod sensitivity determines the flowering time of different varieties of sorghum, and under the condition of long-day temperature zone, many sorghum germplasm sensitive to photoperiod can be flowering late, even without heading, and is not suitable for planting. Since the photoperiod control of flowering was critical to crop yield and hybrid seed production, photoperiod control of flowering has been an important goal of sorghum improvement programs since the beginning of the 20 th century. The traditional widely adaptive sorghum breeding is selected by single plants according to flowering time of breeding offspring, and the method is time-consuming, labor-consuming, easy to be interfered by environment and low in accuracy. The development of specific molecular markers for auxiliary selection is the best method for improving the selection efficiency of the wide-adaptability sorghum by utilizing the base difference of the target genes. Although many scholars have studied the heading stage of sorghum, there are still few reports of genes controlling the heading time of sorghum, and it has been reported that molecular markers associated with the heading stage are more rare. Disclosure of Invention By counting the heading date of the inbred population of Tx430/P898012 in 7 different environments, quantitative Trait Loci (QTLs) were used to map to 5 QTL sites, with the SbFT on Ma1, ma6 and 10 on chromosome 6 being reported and the candidate gene on chromosome 9 being ELF3 and the QTL control gene on chromosome 8 not being reported. The candidate gene on chromosome 8 is cloned into SbDTH < 1 >, and the function and regulation mechanism of SbDTH < 1 > gene are verified and analyzed. According to bioinformatics analysis, sbDTH <1 > codes bHLH transcription factors, the genetic relationship with PIF4 genes of corn is recent, and the SbDTH <1 > protein is found to be located in the cell nucleus through subcellular localization experiments. To analyze SbDTH gene sequence polymorphisms, sequence alignments were performed on Tx430, P898012, tx623, RIO of the published sequences. The early flowering varieties Tx430 and Tx623 in autumn are found to have a 6755bp insertion, while the late flowering varieties P898012 and RIO in autumn are deleted, and are presumed to be functional variations. The transposon is converted into a molecular marker to carry out the statistics of the heading date and the existence of the transposon on an F2 population (Tx 430/red tassel) and a natural population (161 parts of sorghum germplasm), the variety with the transposon is found to be longer than the average heading time of the variety without the transposon, which indicates SbDTH that the sorghum heading date is regulated by 1, and the insertion of the transposon is a functional variation. The invention provides a functional allelic variant in a gene SbDTH for regulating and controlling the heading stage of sorghum, and a transposon is inserted at the 4 th base after a stop codon TGA. Allelic variants may allow for an extended heading time of sorghum. Molecular markers for high throughput marker molecular marker detection have been developed based on allelic variants. The invention is realized by the following technical scheme: the