华北农学报2025，Vol.40Issue(5)：20-25,6.DOI:10.7668/hbnxb.20195855
小麦优质亚基7OE通用双色荧光定量PCR标记的开发
Development of Universal Two-color Fluorescent Quantitative PCR Marker for Wheat Subunit 7OE
摘要
Abstract
Although high-throughput KASP markers have been developed for the wheat quality subunit 7OE,they are different from the KASP markers developed by SNP,the problem of not being able to effectively distinguish between homozygous and heterozygous remains.To clarify the issue of whether the 7OE subunit is homozygous,this study used Jinqiang 6(containing 7OE+8* subunits)and Kenong 199(containing 7+9 subunits)and hybrid off-spring as materials,and used the Waxy-D1 gene of wheat as an internal reference gene.The relative copy number of the 7OE gene to the reference gene was detected by quantitative PCR using the universal dual-color fluorescence used in KASP markers to determine whether the 7OE gene exists and whether it is homozygous,and the detection results were verified by a relevant molecular marker.The results showed that the relative copy number of the parent Jin-qiang 6,with the 7OE gene,was the highest,the relative copy number of the parent Kenong 199,without the 7OE gene,was 0,and the relative copy number of their hybrid F,generation was intermediate,and the three types were easily separated.In its F2 segregating population,the relative copy numbers of the 7OE gene were also easily divided into high,medium and 0 three types.The genotypes that were detected as homozygous and heterozygous for the 7OE gene were further detected by the PCR marker of the 9 subunit(which can detect 9 subunit and the 8* subunit that are closely linked to the 7 subunit and the 7OE subunit,respectively),and the results were completely consistent.The high-throughput 7OE universal dual-color fluorescence quantitative PCR marker established in this study can ac-curately distinguish whether the 7OE subunit is present,and whether it is homozygous,which has a positive effect on promoting the molecular marker-assisted selection of high-quality subunit 7OE.关键词
小麦/优质亚基/7OE/双色荧光/定量PCRKey words
Wheat/High quality subunit/7OE/Dual color fluorescence/Quantitative PCR分类
农业科学引用本文复制引用
刘海晨,周硕,张俊敏,焦博,王娇,董福双,杨帆,赵璞,马春红,柴建芳..小麦优质亚基7OE通用双色荧光定量PCR标记的开发[J].华北农学报,2025,40(5):20-25,6.基金项目
河北省农林科学院现代农业科技创新专项(2022KJCXZX-SSS-4) （2022KJCXZX-SSS-4）
河北省中央引导地方科技发展资金项目(246Z6307G) （246Z6307G）
农业生物育种国家科技重大专项(2023ZD040230303) （2023ZD040230303）
