摘要： 小麦优质亚基7^OE在优质小麦培育方面具有重要作用,虽然针对该优质亚基已开发了高通量的KASP标记,但与由SNP开发的KASP标记不同,仍存在无法有效区分纯合与杂合的问题。为明确7^OE亚基是否纯合的问题,以含有7^OE亚基的津强6号(含7^OE+8^*亚基)和不含7^OE亚基的科农199(含7+9亚基)及其杂交后代为材料,把小麦的Waxy-D1基因作为内参基因,利用KASP标记中使用的通用双色荧光,通过定量PCR检测7^OE基因相对内参基因的相对拷贝数来确定7^OE基因是否存在以及是否纯合,并用相关分子标记对检测结果进行验证。结果表明,含7^OE基因的亲本津强6号相对拷贝数最高,不含7^OE基因的亲本科农199相对拷贝数为0,其杂交F₁相对拷贝数居中,3种类型很容易分开。在其F₂分离群体,7^OE基因的相对拷贝数也很容易分为高、中和0 3种类型,对其中检测为7^OE基因纯合与杂合的基因型进一步用9亚基的PCR标记(能同时检测分别与7亚基和7^OE亚基紧密连锁的9亚基和8^*亚基)进行检测,检测结果完全一致。建立的高通量7^OE通用双色荧光定量PCR标记能准确区分7^OE亚基是否存在以及是否纯合,对促进优质亚基7^OE的分子标记辅助选择具有积极作用。

关键词: 小麦, 优质亚基, 7^OE, 双色荧光, 定量PCR

Abstract:

Although high-throughput KASP markers have been developed for the wheat quality subunit 7^OE,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 7^OE subunit is homozygous,this study used Jinqiang 6(containing 7^OE+8^* subunits)and Kenong 199(containing 7+9 subunits)and hybrid offspring as materials,and used the Waxy-D1 gene of wheat as an internal reference gene.The relative copy number of the 7^OE gene to the reference gene was detected by quantitative PCR using the universal dual-color fluorescence used in KASP markers to determine whether the 7^OE 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 Jinqiang 6,with the 7^OE gene,was the highest,the relative copy number of the parent Kenong 199,without the 7^OE gene,was 0,and the relative copy number of their hybrid F₁ generation was intermediate,and the three types were easily separated.In its F₂ segregating population,the relative copy numbers of the 7^OE gene were also easily divided into high,medium and 0 three types.The genotypes that were detected as homozygous and heterozygous for the 7^OE 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 7^OE subunit,respectively),and the results were completely consistent.The high-throughput 7^OE universal dual-color fluorescence quantitative PCR marker established in this study can accurately distinguish whether the 7^OE subunit is present,and whether it is homozygous,which has a positive effect on promoting the molecular marker-assisted selection of high-quality subunit 7^OE.

Key words: Wheat, High quality subunit, 7^OE, Dual color fluorescence, Quantitative PCR

中图分类号: 

• S512.1小麦