95份甘蔗栽培材料的SSR引物筛选及遗传多样性分析

Analysis of SSR Primers and Genetic Diversity of 95 Sugarcane Culture Materials

【目的】筛选适用于甘蔗主产区种质资源的SSR核心引物,探究中国主要甘蔗材料的遗传多样性和群体结构,为甘蔗亲本材料的保存、利用、甘蔗分子标记辅助选择提供参考。【方法】利用从68对SSR引物中筛选获得的21对核心引物,对采集到的19个群体共95份甘蔗育种亲本材料进行遗传多样性分析。利用Powermarker V3. 25数据处理软件计算引物的多态性信息量(PIC)。根据SSR分子标记数据,计算19个群体的Nei72遗传距离矩阵。利用Powermarker V3. 25软件,对供试材料基于SSR标记的遗传距离矩阵进行相关性分析。根据遗传距离矩阵,利用Mega6. 06软件分别对19个群体进行基于分子标记的类平均法(unweighted pair group method with arithmetic mean,UPGMA)聚类分析。根据SSR分子标记数据,利用STRUCTURE 2. 4对19个群体的95份供试材料进行群体结构分析。【结果】从已公布的68对SSR引物中,筛选获得21对多态性高、稳定性好、综合表现良好的甘蔗SSR分析引物,21对引物中PIC值最大的是0. 9279,最小值为0. 7855,平均值为0. 8664。21对SSR引物共检测出440个甘蔗DNA条带,其中多态性条带数为422个,扩增的DNA条带范围集中在100～500 bp之间。19个群体的Nei72遗传距离最小的为0. 2024,最大的为0. 7262,平均遗传距离为0. 5779,根据UPGMA聚类图可将19个群体划分为4个亚群。群体结构分析表明,当K值等于6时,ΔK取得最大值,说明19个群体的95份材料可以划分为6个亚群,亚群Ⅰ含有4个群体分别为TZ01、GZ01、TZ02、TZ10,亚群Ⅱ含有2个群体为TZ07、TZ08,亚群Ⅲ为TZ11、GZ06、GZ08,亚群Ⅳ分别为GZ07、GZ03、GZ04、GZ05,亚群Ⅴ分别为TZ04、TZ06、TZ05、TZ03、TZ09,亚群Ⅵ只含有一个群体为GZ02。【结论】21对SSR引物能够很好的区分本研究的各个材料,同时供试材料基因组水平的遗传多样性较丰富,可为上述亲本资源的利用奠定良好的基础,并为今后研究中的亲本选择和组合等方面提供重要参考。

【Objective】In this paper,SSR core primers for sugarcane germplasm resources in main producing areas in southern China were selected to explore the genetic diversity and population structure of the main sugarcane materials in southern China,which would provide a reference for the preservation,utilization and selection of sugarcane molecular markers.【Method】The 21 pairs of core primers were selected from 68 pairs of SSR primers,and the genetic diversity analysis of collected 95 sugarcane breeding materials from 19 groups was carried out.Using Powermarker V3.25 data processing software,the polymorphism information of primers(PIC)was calculated.According to SSR molecular marker data,the genetic distance matrix of Nei72 of 19 groups was calculated.Using Powermarker V3.25 software,the correlation analysis of the genetic distance matrix based on SSR marker was carried out.According to the genetic distance matrix,the unweighted pair group method with arithmetic mean based on molecular markers was performed for 19 groups using Mega6.06 software.According to SSR molecular marker data,a group structure analysis was conducted on 95 samples of 19 groups using STRUCTURE 2.4.【Result】From the published 68 pairs of SSR primers,the sugarcane SSR analysis primer was selected to obtain 21 pairs of polymorphic high,stable and well integrated performance,the maximum PIC value of the primers was 0.9279,the minimum value was 0.7855,and the mean value was 0.8664.A total of 440 sugarcane DNA bands from SSR primers were detected,with 422 polymorphisms and the amplification of DNA banding ranges between 100 and 500 bp.The smallest genetic distance of the 19 groups was 0.2024,the largest was 0.7262,the average genetic distance was 0.5779,and 19 groups from UPGMA cluster map could be divided into four subgroups.The analysis of group structure showed,when the K value was equal to the maximumΔK,that the 19 groups of 95 materials could be divided into 6 subgroups:SubgroupⅠcontained four groups for TZ01,GZ01,TZ02,TZ10;SubgroupⅡcontained two groups for TZ07,TZ08;SubgroupⅢcontains for TZ11,GZ06,GZ08;SubgroupⅣcontained for GZ07,GZ03,GZ04,GZ05;SubgroupⅤwas TZ04,TZ06,TZ05,TZ03,TZ09;SubgroupⅥonly contained one group for GZ02.【Conclusion】21 pairs of SSR primers could be used to distinguish the materials in this study,at the same time,the genetic diversity of the genome level was abundant,which could lay a good foundation for the utilization of the above parental resources and the future study of parent selection and combination to provide important reference.