2024年6月19日 星期三
基于SSR分子标记的核桃种质资源分子身份证构建
Establishment of molecular identity cards of walnut (Juglans spp.) germplasm resources based on SSR molecular marker
2023年 第32卷 第2期 页码[1-9]    下载全文[2.4MB]  
摘要

 采用13对SSR引物对54份核桃(Juglans spp.)样本的基因组DNA进行了扩增,对这些引物对应位点的遗传多样性进行了分析;在此基础上,剔除扩增条带缺失超过5%的引物,将剩余引物按照区分率从高到低排序并组合,筛选出能够完全区分供试核桃样本的引物组合;根据筛选出的引物组合中每对引物的扩增结果构建供试核桃样本的分子指纹图谱,并结合各核桃样本的基本信息、选育历史等,利用二维码技术生成每个核桃样本的分子身份证。结果表明:本研究共检测到107个等位基因,供试引物对应位点的观测等位基因数为3~16,均值8.2;有效等位基因数为2.2~7.9,均值4.2;观测杂合度为0.10~0.57,均值0.42;期望杂合度为0.55~0.88,均值0.72;Shannon’s信息指数为0.86~2.36,均值1.60;Nei's基因多样性指数为0.54~0.87,均值0.72;多态信息含量为0.44~0.86,均值0.68。总体来看,引物WGA04和WGA79对应位点的遗传多样性指数较高。剔除引物WGA331、WGA332和WGA376后,其余引物的区分率为5.56%~24.07%。2对引物组合中,WGA04-WGA70的区分率最高(72.22%),以其为基础组成3对引物组合,其中WGA04-WGA70-WGA79、WGA04-WGA70-WGA202、WGA04-WGA70-WGA89和WGA04-WGA70-WGA01的区分率为100.00%。利用WGA04-WGA70-WGA79构建的分子身份证包含各核桃样本的基础信息和简介。研究结果显示:基于SSR分子标记构建的分子身份证可快速、准确鉴定供试核桃种质资源,并可用于核桃种质资源知识产权保护。

Abstract

 The genomic DNA of 54 walnut (Juglans spp.) samples were amplified with 13 pairs of SSR primers, and the genetic diversity of corresponding loci of these primers was analyzed; on the basis, the primers with amplification bands missing more than 5% were discarded, the residual primers were ranked and combined according to the identification rate from high to low, and the primer combinations which can completely identify the test walnut samples were screened; the molecular fingerprints of the test alnut samples were established according to the amplication result of each pair of primers in the screened primer combinations, and the molecular identity card of every walnut sample was established by using two-dimensional code technology combining the basic information, breeding history, etc. of each walnut sample. The results show that 107 alleles are detected in total in this study, and the numbers of observed alleles at the corresponding loci of the test primers are 3-16, with an average of 8.2; the numbers of effective alleles are 2.2-7.9, with an average of 4.2; the observed heterozygosities are 0.10-0.57, with an average of 0.42; the expected heterozygosities are 0.55-0.88, with an average of 0.72; the Shannon's information indexes are 0.86-2.36, with an average of 1.60; the Nei's gene diversity indexes are 0.54-0.87, with an average of 0.72; the polymorphism information contents are 0.44-0.86, with an average of 0.68. Overall, the genetic diversity indexes of the corresponding loci of primers WGA04 and WGA79 are relatively high. After discarding primers WGA331, WGA332, and WGA376, the identification rates of the other primers are 5.56%-24.07%. In combinations of two pairs of primers, the identification rate of WGA04-WGA70 is the highest (72.22%), and combinations of three pairs of primers are formed on this basis, among them the identification rates of WGA04-WGA70-WGA79, WGA04-WGA70-WGA202, WGA04-WGA70-WGA89, and WGA04-WGA70-WGA01 are 100.00%. The molecular identity cards established with WGA04-WGA70-WGA79 contain basic information and introduction of each walnut sample. It is suggested that the established molecular identity cards based on SSR molecular markers can identify the test walnut germplasm resources rapidly and accurately, and they can be used in intellectual property protection of walnut germplasm resources.

关键词核桃; 种质资源; SSR分子标记; 遗传多样性; 分子身份证
Key wordsJuglans spp.; germplasm resource; SSR molecular marker; genetic diversity; molecular identity card
作者马庆国1, 宋晓波1, 贺君星1, 周晔1, 黄勇2, 张俊佩1, 裴东1
所在单位1. 林木遗传育种全国重点实验室 国家林业和草原局林木培育重点实验室 中国林业科学研究院林业研究所, 北京 100091;2. 福建省林业科学研究院, 福建 福州 350012
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基金项目“十四五”国家重点研发计划项目(2019YFD1001603); 福建省属公益类科研院所基本科研专项(2020R1009008); 核桃产业国家创新联盟资助项目(NAWI)