SSR分子标记法快速进行常规稻种子纯度检测的研究及应用  

Research and Application of SSR-based Method for Rapid Seed Purity Assessment of Conventional Rice

作　　者：李儒剑 万吉丽 尹晓佳 王克响 米铁柱 刘佳音[1] 于萌 殷会德 顾晓振 李继明 LI Ru-jian;WAN Ji-li;YIN Xiao-jia;WANG Ke-xiang;MI Tie-zhu;LIU Jia-yin;YU Meng;YIN Hui-de;GU Xiao-zhen;LI Ji-ming(Qingdao Saline-alkali Tolerant Rice Research and Development Center,Qingdao,Shandong 266000,China;Key Laboratory of Marine Environment Science and Ecology of the Ministry of Education,College of Environmental Science and Engineering,Ocean University of China,Qingdao,Shandong 266100,China)

机构地区：[1]青岛海水稻研究发展中心,山东青岛266000 [2]中国海洋大学环境科学与工程学院海洋环境与生态教育部重点实验室,山东青岛266000

出　　处：《杂交水稻》2022年第3期11-19,共9页Hybrid Rice

基　　金：山东省自然科学基金项目(ZR2019QG008);2021年青岛市科技计划科技惠民专项(21-1-4-ny-23-nsh)。

摘　　要：为了提高种子纯度鉴定效率,实现常规稻种子纯度的快速检测,本研究首先对几种简化DNA提取试剂和操作方法进行比较,结果表明以Buffer A、Buffer B作提取试剂,在提取过程中对种子充分研磨并进行95℃水浴,提取的DNA进行PCR,电泳条带均一且明亮,适用于SSR标记扩增。另外,利用多态性标记检测种子纯度时,通过5粒、10粒种子进行DNA混样,筛选能够扩增出差异条带的标记用于后续单株检测,筛选出的标记位点对待检种子的检测结果与前期标记筛选一致。因此,通过5粒、10粒种子的DNA混样初筛,而后对扩增出差异条带的混样内包含的单株进行鉴定能够准确定位到假种子,进而实现种子纯度的检测。这种方法在节约试剂、降低检测成本的同时,缩短了检测时间,大大提高了种子纯度检测效率。In order to increase the efficiency of seed purity assessment of conventional rice,several simplified DNA extraction methods were compared in this study.The results showed that the DNA extraction procedure using Buffer A and Buffer B as extraction reagents and seeds being fully ground and inoculated in a 95℃water bath was optimal for SSR marker detection.The PCR amplified bands of DNA templates extracted by this method were uniform and bright.In addition,the differential PCR band of the fake seed was still distinct when the DNA was mixed in 1/5 up to 1/10ration.During the screening of polymorphic markers for seed purity test,the markers that amplified different bands in these DNA pools were selected for further screening using DNA of a single plant.Polymorphic markers were identified in all individuals and the results were consistent with the marker screening using DNA pools.Therefore,by mixing the DNA of five seeds or ten seeds,a primary screening can be performed to identify the markers for the polymorphism,and narrow down to the groups of samples that contain fake seed and further accurately identify the single plant of fake seed.Not only does this method save reagents and reduce cost,but also shortens detection time and greatly improves the efficiency of seed purity assessment.

关 键 词：常规稻 SSR分子标记 种子纯度 快速检测 

分 类 号：S511.01[农业科学—作物学] S311