96份中、日春兰资源遗传多样性的ISSR分析  

Genetic Diversity ISSR Analysis of 96 Chinese and Japanese Resources of Cymbidium goeringii Based on ISSR Markers

作　　者：孙叶[1,2] 赵国琦[1] 袁媛[2] 张甜[2] 刘红[2] 曹宏[2] 包建忠[2] 陈秀兰[2] Sun Ye;Zhao Guoqi;Yuan Yuan;Zhang Tian;Liu Hong;Cao Hong;Bao Jianzhong;Chen Xiulan(College of Animal Science and Technology,Yangzhou,225009;Institute of Agricultural Sciences of Lixiahe Districts,Yangzhou,225007)

机构地区：[1]扬州大学动物科学与技术学院,扬州225009 [2]江苏里下河地区农业科学研究所,扬州225007

出　　处：《分子植物育种》2020年第5期1535-1547,共13页Molecular Plant Breeding

基　　金：江苏省“六大人才高峰”高层次人才项目(NY-139);江苏省农业科技自主创新资金(CX(17)2012);江苏省农业种质资源保护与利用平台项目(JSGB2018-1)共同资助。

摘　　要：为了探讨中、日春兰优良种质资源的生物遗传多样性,本研究通过L9(34)正交试验,确立ISSR-PCR最佳扩增反应体系,筛选ISSR引物对4个类群96份中、日春兰种质材料进行标记扩增。结果表明:本试验筛选出的ISSR-PCR最佳扩增反应体系中主要影响因子的浓度为:Mg2+浓度2 mmol/L,引物浓度0.4μmol/L,Taq聚合酶用量1.5 U/20μL,d NTPs浓度0.2μmol/L;筛选出的13条引物在93份材料中扩增出126个条带,每条引物平均扩增条带数为9.69,多态性比例(PPB)为100%,种群总等位基因数(Na)为2.000 0,有效等位基因数(Ne)为1.304 5,Nei’s基因多样性(He)为0.203 3,Shannon’s信息多样性指数(Ⅰ)为0.334 0,种群水平上的遗传多样性较高;在类群水平上PPB平均值为82.94%,Na平均值为1.829 4,Ne平均值为1.301 1,He平均值为0.193 8,Ⅰ平均值为0.311 9;类群间的遗传分化系数(Gst)为0.052 1,基因流水平(Nm)为9.089 6,类群间的基因交流程度很高,类群间的变异低于类群内的变异;各类群的遗传多样性水平由高到低依次为Ⅰ类群(中国春兰正格花品种)>Ⅲ类群(中国春兰杂交种)>Ⅱ类群(中国春兰蝶花品种)>Ⅳ类群(日本春兰品种),4个春兰类群两两之间的遗传相似性系数(GS)范围为0.966 1~0.995 1,总体上遗传距离较小。UPGMA聚类分析结果显示,ISSR分子标记能很好的鉴定表型性状相似的春兰原生种和杂交种,在遗传相似系数为0.82时,Ⅱ类(中国春兰蝶花品种)品种和Ⅳ类(日本春兰品种)品种能分组聚类,Ⅰ类(中国春兰正格花品种)品种和Ⅲ类(中国春兰杂交种)种质混合在一起。春兰是中国的传统特色花卉,春兰优良品种的产业化开发潜力大,中、日春兰种质资源的遗传多样性研究对春兰种质的保护、利用和创新有重要的意义。In order to analyze the genetic diversity among 96 germplasms of C. goeringii from China and Japan,through the L9(34) orthogonal experiment, the ISSR-PCR marker system was established, ISSR primers were screened and used to amplify the markers on 96 resources of C. goeringii divided into four groups. The results showed that the concentration on which were the main influence factors of ISSR-PCR system of this test contained 2 mmol/L Mg2+, 0.4 μmol/L primer, 1.5 U/20 μL Taq polymerase, 0.2 μmol/L d NTPs;13 ISSR primers were screened and126 bands were amplified from 93 samples, the percentage of polymorphic bands was 100%, the genetic diversity of germplasms at species level was higher(PPB=100%, Na=2.000 0, Ne=1.304 5, He=0.203 3, I=0.334 0). The genetic diversity of four groups showed the results(PPB=82.94%, Na=1.8294, Ne=1.301 1, He=0.193 8, I=0.311 9, the Nei’s coefficient differetiations(Gst) was 0.052 1, the estimate of gene flow(Nm)was 9.089 6, these indicated that genetic differentiation in the groups was higher among the groups. The genetic differentiation in the groups was as follows from large to small: typeⅠgroups(wild-type cultivars of C. goeringii from China)>typeⅢ(hybrid germplasms of C. goeringii from China)>typeⅡ(peloric-mutant cultivars of C. goeringii from China)>typeⅣ(C. goeringii cultivars from Japan). The genetic similarties coefficient(GS) between groups ranged from 0.966 1 to 0.995 1, suggesting the genetic distance between groups was closed. UPGMA clustering analysis showed that ISSR molecular markers could be used to distinguish primary cultivars and hybrid germplasms of C. goeringii which Phenotypic traits were similar, at a similarity coefficient criterion of 0.82, peloric-mutant cultivars of C. goeringii from China and C. goeringii cultivars from Japan were clustered to partitions respectively, and primary wild-type cultivars and hybrid germplasms of C. goeringii from China were mixed up together. C. goeringii is Chinese traditional characteristic flowers, the industrial deve

关 键 词：春兰(Cymbidium goeringii) 遗传多样性 ISSR 聚类分析 

分 类 号：S682.31[农业科学—观赏园艺]