基于两种电泳检测方法的华东野生菰ISSR遗传多样性分析  

Genetic diversity analysis of Zizania latifolia by ISSR in East China based on two electrophoresis detection methods

作　　者：谢小燕 苏晓娜[2] 王惠梅[3] 江绍琳[4] 吴建利[3] 江绍玫[5] XIE Xiao-yan;SU Xiao-na;WANG Hui-mei;JIANG Shao-lin;WU Jian-li;JIANG Shao-mei(College of Art,Jiangxi University of Finance and Economics,Nanchang 330013,China;Nanchang Business College of Jiangxi Agricultural University,Nanchang 330013,China;China National Rice Research Institute,Hangzhou 310006,China;Agronomy College,Jiangxi Agricultural University,Nanchang 330045,China;College of Statistics,Jiangxi University of Finance and Economics,Nanchang 330013,China)

机构地区：[1]江西财经大学艺术学院,南昌330013 [2]江西农业大学南昌商学院,南昌330013 [3]中国水稻研究所,杭州310006 [4]江西农业大学农学院,南昌330045 [5]江西财经大学统计学院,南昌330013

出　　处：《南方农业学报》2019年第12期2638-2646,共9页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31460378);国家转基因生物新品种培育重大专项(2016ZX08001-002)

摘　　要：【目的】筛选更优的ISSR-PCR扩增产物电泳检测方法,分析我国华东地区野生菰种质资源遗传多样性,为建立和完善菰植物种质库及合理开发与利用菰资源提供理论依据。【方法】挑选7条ISSR引物对华东地区9个野生菰居群104份材料基因组DNA进行PCR扩增,其ISSR-PCR产物分别采用2%琼脂糖凝胶电泳和5%非变性聚丙烯酰胺凝胶电泳进行检测。【结果】2%琼脂糖凝胶电泳检测结果显示,华东地区野生菰的多态性位点百分率(PPB)为63.49%、观测等位基因数(Na)为1.6349、有效等位基因数(Ne)为1.3012、Nei’s基因多样性指数(He)为0.1871、Shannon’s信息指数(I)为0.2908、居群间遗传分化系数(Gst)为0.3922、遗传相似系数(Gs)变化范围在0.8293~0.9775;其聚类分析结果表明,当遗传相似性系数阈值为0.885时,9个野生菰居群被分为两大类,SH(上海)和PYH(鄱阳湖)居群归为一类,其他7个居群归为另一类。5%非变性聚丙烯酰胺凝胶电泳检测结果显示,华东地区野生菰的PPB为78.03%、Na为1.7803、Ne为1.2762、He为0.1804、I为0.2914、Gst为0.5086、Gs变化范围在0.7679~0.9837;其聚类分析结果表明,当遗传相似性系数阈值为0.850时,9个居群也被分为两大类,其中PYH居群单独为一类,其他8个居群归为另一类。【结论】我国华东地区野生菰种质资源遗传多样性丰富,居群间和居群内变异显著,基于ISSR分子标记的野生菰遗传多样性采用5%非变性聚丙烯酰胺凝胶电泳进行检测可获得更可靠的遗传图谱,而2%琼脂糖凝胶电泳检测更适合早期的ISSR分子标记引物筛选。【Objective】To select better electrophoretic detection methods for ISSR-PCR amplification products,analyze the genetic diversity of Zizania latifolia germplasm resources in East China,and provide theoretical basis for the establishment and improvement of Z. latifolia germplasm banks and the rational development and utilization of the resources.【Method】Seven ISSR primers were screened out to amplify 104 genomic DNA from 9 Z. latifolia populations in East China. The ISSR-PCR products were analyzed by 2% agarose gel and 5% non-denaturing polyacrylamide gel detection.【Result】The results of agarose gel electrophoresis showed that the percentage of polymorphic loci(PPB)of Z. latifolia in East China was 63.49%,the number of observed alleles(Na)was 1.6349,and the number of effective alleles(Ne)was1.3012,Nei’s genetic diversity index(He)was 0.1871,Shannon information index(I)was 0.2908,genetic differentiation coefficient(Gst)between populations was 0.3922,and genetic similarity coefficient(Gs)varied from 0.8293 to0.9775. The results of cluster analysis showed that when the threshold of genetic similarity coefficient was 0.885,the nine Z. latifolia populations were classified into two categories,the SH(Shanghai)and PYH(Poyang Lake)populations were classified into one group,and the other seven populations were classified into another group. The results of 5% non-denaturing polyacrylamide gel electrophoresis showed that the PPB of Z. latifolia in East China was 78.03%,Na was 1.7803,Ne was 1.2762,He was 0.1804,I was 0.2914,Gst was 0.5086,and Gs ranged from 0.7679 to 0.9837. The results of cluster analysis showed that when the threshold of genetic similarity coefficient was 0.850,the nine populations were also divided into two categories,among which the PYH population was classified into one category and the other eight populations were classified into another category.【Conclusion】The Z. latifolia germplasm resources in East China are rich in genetic diversity,with significant variation between and within popu

关 键 词：野生菰 ISSR分子标记 遗传多样性 琼脂糖凝胶电泳 聚丙烯酰胺凝胶电泳 华东地区 

分 类 号：S519[农业科学—作物学] Q943[生物学—植物学]