Search

CN-120989282-B - Molecular marker of locus qSGC.A8-1 obviously related to brassica napus seed thio glycoside content and application thereof

CN120989282BCN 120989282 BCN120989282 BCN 120989282BCN-120989282-B

Abstract

The invention belongs to the technical fields of molecular biology and genetic breeding, and discloses a molecular marker of a locus qSGC.A8-1 obviously associated with brassica napus seed thio glycoside and application thereof. The locus qSGC.A8-1 and the peak SNP marker Bn-A08-p2660414 thereof which are obviously related to the content of the glucosinolate of the rape seed are positioned at the 2,088,727 th base of the A08 chromosome of the rape DarmorV4.1 reference genome, the average can explain 8.8 percent of phenotype variance, and the average additive effect is-12.0 mu mol/g. The PARMS mark designed by the SNP is used for detecting rape germplasm resources, and the detection shows that the operation is simple and the typing is clear, and the average sulfan content of the A genotype is lower than that of the G genotype type material seeds, so that the method has a good selection effect on rape seed sulfan.

Inventors

  • SHI JIAQIN
  • WANG HANZHONG
  • WANG XINFA
  • DUN XIAOLING
  • WU XUEKE
  • Du Tianhang

Assignees

  • 中国农业科学院油料作物研究所

Dates

Publication Date
20260508
Application Date
20250820

Claims (7)

  1. 1. The application of the reagent for detecting 2,088,727 th base on the brassica napus A08 chromosome in brassica napus seed thio-glycoside content screening breeding is characterized in that the determination mode in the application process is that if the 2,088,727 th base on the brassica napus A08 chromosome is detected to be G, the content of thio-glycoside in the brassica napus seed is high, and if the 2,088,727 th base on the brassica napus A08 chromosome is detected to be A, the content of thio-glycoside in the brassica napus seed is low, and the version number of the brassica napus genome is B.napus Darmor-bzh reference genome sequence assembly version 4.1.1.
  2. 2. The application of the reagent for detecting 2,088,727 th base on the brassica napus A08 chromosome in preparing brassica napus seed thio-glycoside content screening reagent kit is characterized in that the method is that if 2,088,727 th base on the brassica napus A08 chromosome is detected to be G, the content of thio-glycoside in the brassica napus seed is high, and if 2,088,727 th base on the brassica napus A08 chromosome is detected to be A, the content of thio-glycoside in the brassica napus seed is low, and the version number of the brassica napus genome is B.napus Darmor-bzh reference genome sequence assembly version 4.1.1.
  3. 3. The use according to claim 1 or 2, wherein the agent is a primer.
  4. 4. The use according to claim 3, wherein the primer is PARMS detection primer.
  5. 5. The method of claim 4, wherein the PARMS detection primers are qSGC.A8-1F, CACCGTACTTCTCCATCC, qSGC.A8-1Rt, gaaggtgaccaagttcatctTCCAATATGGTTGGTAGTAAGAT, and qSGC.A8-1Rc, gaaggtcgagtcaacggattTCAATAGTGGTGGTAGTAAGAC.
  6. 6. A method for screening and breeding the content of the thio glycoside in the seeds of brassica napus includes such steps AS detecting the 2,088,727 th basic group on the A08 chromosome of brassica napus, sequencing, taqMan probe method, AS-PCR method, molecular beacon method, high-resolution dissolving curve method, CAPS method, snapShot method, KASP method, PARMS method, gene chip method or mass spectrum method, and features that if the 2,088,727 th basic group on the A08 chromosome of brassica napus is G, it is indicated that the content of thio glycoside in the seeds of brassica napus is high, and if the 2,088,727 th basic group on the A08 chromosome of brassica napus is A, it is indicated that the content of thio glycoside in the seeds of brassica napus is low, and the version number of genome of brassica napus is B.napus Darmor-bzh reference genome sequence assembly version 4.1.1.
  7. 7. The method of claim 6, wherein the detection method is PCR using primers qSGC.A8-1F, CACCGTACTCTCTCCATCC, qSGC.A8-1RT, gaaggtgaccaagttcatctTCCAATAGTGGTGGTAGATTAAGAT and qSGC.A8-1RC gaaggtcggtagtcaacggattTCAATAGGTGGTTGGTAGATTAAAAC.

Description

Molecular marker of locus qSGC.A8-1 obviously related to brassica napus seed thio glycoside content and application thereof Technical Field The invention belongs to the technical field of biological breeding, and particularly relates to a molecular marker of a locus qSGC.A8-1 obviously related to the content of brassica napus seed thioglycoside and application thereof. Background Currently, the main goals of rape breeding include resistance, yield and quality. These traits are complex quantitative traits, are commonly regulated and controlled by a plurality of genes and are easily influenced by environmental conditions, and the precise improvement of these traits is difficult to realize by traditional breeding methods and technical means. With the rapid progress of molecular marking technology and the wide application of emerging biotechnology such as gene editing, the molecular improvement of these traits is possible. The quality traits of rape mainly include oil content of rape seed, erucic acid and thioglycoside (thioglucoside) content, etc. Thioglycoside is a sulfur-containing secondary metabolite, and the core structure consists of beta-thioglucosyl, a sulfonyl oxime group and a side chain R group from different amino acids. The thioglycoside is mainly present in cruciferous plants such as rape, broccoli, cauliflower, etc. Thioglycoside has various positive effects in cancer prevention in humans, plant pest defense, imparting a specific flavor to vegetables, and the like (Xieqi, 2023). However, the thioglycoside in the rapeseed cake has a toxic effect on the farm animals, so the value of the rapeseed cake as a feed is determined by the content of the thioglycoside in the rapeseed grains. By means of linkage and/or associative localization, a number of QTLs have been identified in canola (Chao et al, 2022; zhang et al, 2024) controlling seed sulfan content, mostly distributed on the A9, C2, C7 and C9 chromosomes, mainly corresponding to genes such as GTR2 and MYB28 (Zhang et al, 2024). However, these QTLs are insufficient to account for variation in the level of glucosinolates in the germplasm resources of canola, indicating that there is still a need for new control sites for the level of glucosinolates. The invention utilizes the high-density SNP genotype data of rape core association groups and the seed sulfan content phenotype data of 8 environments to carry out whole genome association analysis, aims at finding new stable association sites, and develops practical high-throughput low-cost molecular markers for molecular improvement of the rape seed sulfan content. Disclosure of Invention The invention aims at providing an application of a reagent for detecting 2,088,727 th basic group on a cabbage type rape A08 chromosome in screening and breeding of cabbage type rape seed thio-glycoside content. The invention also aims at providing an application of the primer for detecting 2,088,727 th basic group on the A08 chromosome of the brassica napus in the screening breeding of the brassica napus seed thio-glycoside content. The final object of the invention is to provide a method for screening and breeding brassica napus seed thio-glycoside content. In order to achieve the above object, the present invention adopts the following technical measures: obtaining a PARMS marker with obvious association of the brassica napus seed thio glycoside content: (1) The total DNA of 331 parts of materials (Li et al, 2020) of the cabbage type rape core association group constructed by the team is extracted, and each sample is subjected to genotype analysis by using a rape 60K SNP chip (Clarke et al, 2016). (2) Marker heterozygosity (heterozygous rate), deletion rate (MISSING RATE), minimum allele frequency (minor allele frequency) of the population material at each locus was calculated using Illumina BeadStudio genotyping software (http:// www.illumina.com /). And removing the markers with no polymorphism, high deletion proportion, no homozygous genotype, low allele frequency, high heterozygosity genotype frequency, uncertain positions and multiple copies, and finally obtaining 24508 high-quality SNP markers for subsequent analysis. (3) 331 Parts of materials of the cabbage type rape core association group constructed by the team are planted in eight environments (code numbers are N14, W12, W13, W14, W15, W16, Z13 and Z14) of Nanchang 2014, wuhan 2012-2016 and Zhengzhou 2013-2014 respectively, 10 plants are harvested in each cell in the mature period, aired and threshed, and the content of the thioglycoside is measured by a near infrared method (Qia et al, 2006). (4) Association analysis was performed using TASSEL 5.0.0 software (braddury et al, 2007) in combination with SNP genotype, seed-sulfan content phenotype, population structure and relationship data for the core association population. Finally, a locus qSGC.A8-1 which is obviously related to various sulfas is obtained on a rape DarmorV reference genome A08 chromo