中国棉花审定品种SSR指纹库的构建与综合评价  

Construction of SSR Fingerprint Library and Comprehensive Evaluation for Approved Cotton Varieties in China

作　　者：吴玉珍 黄龙雨 周大云[1,2] 黄义文 付守阳 彭军 匡猛[1,2] WU YuZhen;HUANG LongYu;ZHOU DaYun;HUANG YiWen;FU ShouYang;PENG Jun;KUANG Meng(Institute of Cotton Research,Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology,Anyang 450001,Henan;National Nanfan Research Institute(Sanya),Chinese Academy of Agricultural Sciences,Sanya 572024,Hainan)

机构地区：[1]中国农业科学院棉花研究所/棉花生物育种与综合利用全国重点实验室,河南安阳455000 [2]三亚中国农业科学院国家南繁研究院,海南三亚572024

出　　处：《中国农业科学》2024年第8期1430-1443,I0001-I0061,共75页Scientia Agricultura Sinica

基　　金：三亚崖州湾科技城科技专项(SCKJ-JYRC-2022-62);棉花生物育种与综合利用全国重点实验室项目(CB2022C08);农业生物育种重大项目(2022ZD04019);海南省自然科学基金联合项目(SQ2021ZRLH0113)。

摘　　要：[目的]棉花是一种异源四倍体作物,基因组结构复杂,常异花授粉的繁殖方式也导致棉花品种难以实现高度纯合;棉种市场缺乏有效的监管技术手段,品种多乱杂现象长期存在,严重影响纤维品质一致性。构建中国近20年棉花审定品种标准样品的DNA指纹库,探索棉花品种高通量SSR身份鉴定模式,为棉花品种真实性鉴定和新品种特异性鉴定提供依据;分析审定品种的遗传多样性和群体分化,为棉花不同生态区的适应性鉴定和培育适应新环境的品种提供理论基础。[方法]基于多重PCR技术和毛细管电泳检测方法,使用筛选得到的60个SSR标记构建1 015份棉花审定品种标准样品的DNA指纹库,通过植物品种DNA指纹库管理系统对审定品种SSR指纹进行两两比对,分析审定品种的遗传差异,筛选用于品种鉴定的核心SSR位点。利用聚类分析法和群体结构分析法分析1 015份棉花审定品种的遗传多样性,并计算群体间遗传分化指数。[结果]60个SSR标记在1 015个审定品种中共扩增出216种等位变异,平均等位变异数为3.6,平均PIC值为0.37。1 015个审定品种的SSR指纹进行两两比较,共产生513 591组结果,样品间最大差异位点数为58个。差异位点百分比主要集中在41%-70%,涉及428 115组,占83.36%;其中,差异位点百分比在51%-60%时,涉及组数最多,为197 829组,占38.52%。品种间差异位点百分比大于20%时,占所有品种两两比对组数的99%以上,差异位点百分比低于20%的比对结果只占0.58%。基于组合鉴定法,筛选一套包含10个SSR位点的核心位点组合,在1 015个品种中鉴别能力达到99%。聚类结果和群体结构分析表明,1 015个品种被清晰地划分为5个亚群,G1(n=240)为早熟棉亚群,主要分布于中国北部和西北内陆地区,该亚群品种遗传多样性最丰富,品种间平均遗传距离为0.419。G2(n=277)亚群为中熟棉亚群,分布于长江流域,该亚群杂交种较多,亚群�[Objective]Cotton,a heterotetraploid crop with a complex genome structure,faces challenges in achieving high homozygosity due to frequent cross-pollination.The absence of effective technical supervision in the cotton seed market and the persistence of disordered varieties have a negative impact on the consistency of fiber quality.The objectives of this study are threefold:to establish a DNA fingerprint database for approved cotton varieties in China over the past 20 years,to explore a high-throughput SSR identification model for cotton varieties,and to provide a basis for the authentication of existing varieties and the specific identification of new cotton varieties.Additionally,we aim to analyze the genetic diversity and population differentiation among approved varieties.Ultimately,our goal is to provide a theoretical framework for identifying cotton varieties that are well-suited to different ecological regions and for developing varieties that can adapt to new environments.[Method]Based on multiplex PCR technology and capillary electrophoresis detection method,using 60 SSR markers screened to construct a DNA fingerprint library of 1015 standard samples of cotton approved varieties.Through the plant variety DNA fingerprint library management system,the SSR fingerprints of approved varieties were compared pairwise to analyze the genetic differences of approved varieties and screen the core SSR loci for variety identification.Cluster analysis and population structure analysis were used to analyze the genetic diversity of 1015 cotton approved varieties and calculate the genetic differentiation index between populations.[Result]60 SSR markers amplified 216 allelic variations in 1015 approved varieties,with an average of 3.6 allelic variations and a mean PIC value of 0.37.When the SSR fingerprints of the 1015 approved varieties were compared,a total of 513591 pairwise results were generated,with a maximum of 58 different loci between samples.The percentage of different loci was mainly concentrated at 41%-70%,invol

关 键 词：棉花 标准样品 SSR标记 DNA指纹库 综合评价 

分 类 号：S562[农业科学—作物学]