基于SSR的陕西糜子种质资源的分子鉴定  

Molecular identification of broomcorn millet germplasm resources in Shaanxi based on SSR

作　　者：郭娟 辛旭霞 冯智尊 曹越 王晓丹[1] 曹晓宁 SANTRA Dipak K 陈凌[2] 乔治军 王瑞云[1,2] GUO Juan;XIN Xu-Xia;FENG Zhi-Zun;CAO Yue;WANG Xiao-Dan;CAO Xiao-Ning;SANTRA Dipak K;CHEN Ling;QIAO Zhi-Jun;WANG Rui-Yun(Agronomy College,Shanxi Agricultural University,Taigu 030801,Shanxi,China;Center for Agricultural Genetic Resources Research,Shanxi Agricultural University/Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau,Ministry of Agriculture and Rural Affairs/Shanxi Key Laboratory of Genetic Resources and Genetic Improvement of Minor Crops,Taiyuan 030031,Shanxi,China;Panhandle Research&Extension Center,Department of Agronomy and Horticulture,University of Nebraska-Lincoln,Scottsbluff 69361,Nebraska,USA)

机构地区：[1]山西农业大学农学院,山西太谷030801 [2]山西农业大学农业基因资源研究中心/农业农村部黄土高原作物基因资源与种质创制重点实验室/杂粮种质资源发掘与遗传改良山西省重点实验室,山西太原030031 [3]内布拉斯加大学林肯分校农艺系小宗粮豆研究与推广中心,美国内布拉斯加州斯克茨布拉夫69361

出　　处：《作物学报》2024年第10期2643-2653,共11页Acta Agronomica Sinica

基　　金：国家农作物种质资源平台项目(NCGRC-2024-026);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-06-14.5-A16);山西省现代农业产业技术体系建设(杂粮)项目(2024CYJSTX0312);山西省重点研发项目(2022ZDYF110);山西农业大学农学院研究生教育改革与质量提升工程项目(2023YCX33,2023YCX48,2023YCX51,2023YDT05)资助。

摘　　要：为构建糜子(Panicum miliaceum L.)DNA分子身份证,试验以181份陕西糜子核心种质为材料,对课题组前期开发的糜子特异性SSR标记进行多次PCR筛选和优化后获取核心引物。基于糜子参考基因组信息,经过BLAST序列比对后将核心标记进行染色体定位。在SSR引物的5′端标注荧光(FAM/HEX),根据毛细管电泳检测的片段有无采用“0/1”表示,利用ID Analysis4.0进行区分,十进制(0~9)统计扩增片段大小构建材料字符串,用Pop Gene、Power Marker、MEGA、Structure和NTSYS进行遗传多样性分析。试验结果表明,7个荧光SSR(RYW3、RYW6、RYW37、RYW40、RYW43、RYW125和RYW146)组合可区分181份材料,不均匀的分布在5条染色体上,共检测出77个等位变异,平均为11;检出Shannon多样性指数(I)为0.8145(RYW146)~7.8254(RYW125),平均5.9076;观测杂合度(Ho)为0.2627(RYW146)~0.9506(RYW3);期望观测杂合度(He)为0.3329(RYW146)~0.8747(RYW125);Nei’s基因多样性指数(Nei)为0.3315(RYW146)~0.8722(RYW125);多态性信息含量(PIC)为0.5923(RYW146)~0.9445(RYW125),平均为0.8419。基于UPGMA将181份资源划分为3个群组。基于主成分分析将材料分为10个类群,与地理来源一致。利用在线二维码技术(https://cli.im/)构建181份陕西糜子核心种质的DNA分子身份证。To construct the DNA molecular identity card of broomcorn millet(Panicum miliaceum L.),we conducted an experiment using 181 core germplasm samples of Shaanxi millets.The core primers were obtained through several rounds of PCR screening and optimization of millet-specific SSR markers developed in the preliminary stage of our research group.Chromosomal localization of the core markers was performed by comparing their sequences with the reference genome information of broomcorn millet using BLAST.FAM/HEX labels were added the 5'end of SSR primers,and the presence or absence of fragments detected by capillary electrophoresis was represented as“0/1”.The differentiation of fragments was carried out using ID Analysis 4.0,and the size of the amplified fragments was recorded in format(0-9)to construct unique material strings.These strings were then subjected to genetic diversity analysis using software tools such as Pop Gene,Power Marker,MEGA,Structure,and NTSYS.Our test results showed that seven combinations of fluorescent SSRs(RYW3,RYW6,RYW37,RYW40,RYW43,RYW125,and RYW146)could distinguish the 181 materials.These markers were unevenly distributed across five chromosomes,and a total of 77 allelic variants were detected,with an average of 11 variants per marker.The observed Shannon's diversity index(I)ranged from 0.8145(RYW146)to 7.8254(RYW125),with an average of 5.9076.The observed heterozygosity(Ho)ranged from 0.2627(RYW146)to 0.9506(RYW3),while the expected heterozygosity(He)ranged from 0.3329(RYW146)to 0.8747(RYW125).Nei’s gene diversity index(Nei)varied from 0.3315(RYW146)to 0.8722(RYW125).The Polymorphism Information Content(PIC)ranged from 0.5923(RYW146)to 0.9445(RYW125),with an average of 0.8419.Using the UPGMA clustering method,we categorized the 181 resources into three clusters.Principal component analysis resulted in the classification of the materials into ten taxa,which were consistent with their geographic sources.To provide convenient access to the DNA molecular identity cards,we utilized online QR

关 键 词：糜子 陕西 荧光SSR 毛细管电泳 DNA分子身份证 

分 类 号：S516[农业科学—作物学]