油莎豆SRAP-PCR体系优化及遗传多样性分析  

Optimization of SRAP-PCR System and Genetic Diversity Analysis of Cyperus esculentus

作　　者：赵琦琦 郭玉静 于梦斐 王颖[4] 高文伟[1] 张斌[2] Zhao Qiqi;Guo Yujing;Yu Mengfei;Wang Ying;Gao Wenwei;Zhang Bin(College of Agronomy,Xinjiang Agricultural University,Urumqi 830000,China;Institute of Crop Germplasm Resources,Shandong Academy of Agricultural Sciences,Jinan 250100,China;Longping Branch,Graduate School of Hunan University,Changsha 410125,China;College of Life Sciences,Qufu Normal University,Qufu 273165,China)

机构地区：[1]新疆农业大学农学院,新疆乌鲁木齐830000 [2]山东省农业科学院农作物种质资源研究所,山东济南250100 [3]湖南大学研究生院隆平分院,湖南长沙410125 [4]曲阜师范大学生命科学学院,山东曲阜273165

出　　处：《山东农业科学》2022年第8期31-38,共8页Shandong Agricultural Sciences

基　　金：国家重点研发计划项目“黄河三角洲耐盐碱作物良种选育关键技术与规模化制种”(2019YFD1002701);“重要耐盐碱作物规模化种植丰产增效技术集成与示范”(2019YFD1002703);山东省“渤海粮仓”科技示范工程项目(2019BHLC002)。

摘　　要：以16份油莎豆种质资源为材料,用改良的CTAB法提取基因组DNA,利用单因素试验和正交试验研究引物浓度、混合酶体积和DNA模板含量3个因素对油莎豆SRAP-PCR扩增结果的影响,以优化其SRAP-PCR体系,并对供试油莎豆种质材料的遗传多样性进行分析。结果表明,混合酶体积对SRAP-PCR反应体系影响最大,引物浓度影响最小;油莎豆SRAP-PCR总体系为15μL时,引物浓度为0.5μmol/L、混合酶体积为10.5μmol/L、DNA模板含量为80 ng的扩增效果最佳。利用最佳扩增体系,从102对引物组合中筛选出30对多态性引物,共扩增出289个多态性位点,平均多态性比率为92.82%。进一步对16份油莎豆种质的遗传多样性进行分析,结果显示其遗传一致度范围为0.60~0.93,遗传距离范围为0.07~0.40,表明供试油莎豆种质资源的遗传背景差异较小;UPGMA聚类结果表明,油莎豆种质资源亲缘关系受油莎豆品种特性和地域的影响。本研究结果可为油莎豆种质资源的多样性评价及鉴定提供方法和技术,并为其遗传育种提供依据。Using 16 germplasm resources of Cyperus esculentus as materials,the genomic DNA was extracted by improved CTAB method.The single-factor and orthogonal experiments were used to study the effects of primer concentration,mixed enzyme volume and DNA template content on the amplification results of SRAP-PCR in order to optimize the reaction system,and then the genetic diversity of the tested C.esculentus germplasms were analyzed.The results showed that the mixed enzyme volume had the greatest influence on SRAP-PCR reaction system,and the primer concentration had the least influence.When the total volume of SRAP-PCR system was 15 μL,the optimal amplification results could obtained at 0.5 μmol/L primer,10.5 μmol/L mixed enzyme and 80 ng DNA template.Using the optimal amplification system,30 pairs of polymorphic primers were screened out from 102 pairs,and 289 polymorphic sites were amplified with an average polymorphism ratio of 92.82%.The genetic concordance ranged from 0.60 to 0.93,and the genetic distance ranged from 0.07 to 0.40,indicating that there were fewer differences in genetic background of the tested C.esculentus germplasms.The UPGMA cluster results showed that the kinship of C.esculentus germplasm resources was affected by species characteristics and territorial influence.These results could provide method and technique for genetic diversity evaluation and identification of C.esculentus germplasm resources,and bases for further genetic breeding.

关 键 词：油莎豆 SRAP-PCR 扩增体系优化 遗传多样性 

分 类 号：S565.903[农业科学—作物学]