| 摘要: |
| 【目的】评判限制性位点扩增多态性（RSAP）标记对杨树无性系良种的鉴定效果,为杨树种质资源保护与品系鉴定提供依据。【方法】以白杨派10个优良无性系（秦白杨1号、秦白杨3号、秦白杨5号、84K、I-101、新疆杨、毛白杨30号、07-17-18、07-23-23、07-30-11）为材料，从9条RSAP引物组成的36对引物组合中筛选出6对扩增效果较好的引物组合，利用毛细管电泳检测法及POPGENE 32软件检测并分析扩增产物多态性，计算遗传多样性相关指标；选取多态性高且能区分所有无性系的引物，构建白杨派10个无性系的指纹图谱；利用NTSYS 2.1软件计算各无性系间的遗传相似系数，基于UPGMA进行聚类分析。【结果】6对引物组合从10个白杨派无性系中共扩增多态位点133个，平均每对引物扩增22.17个，平均Shannon信息指数为0.326，平均Nei’s基因多样性指数为0.488，表明无性系间具有较高的遗传多样性，遗传变异丰富。利用2个引物组合Rs2/Rs4和Rs2/Rs7的6个多态性位点，即可将10个白杨派无性系全部鉴别开来。10个白杨派无性系间的遗传相似系数为0.400~0.817，平均遗传相似系数为0.594。聚类结果显示，在遗传相似系数0.510处，10个白杨派无性系被分成两大类，新疆杨和毛白杨30号聚为一类，其余无性系聚为另一类，与其实际亲缘关系大致相符。【结论】RSAP分子标记可有效用于杨树种质资源鉴定及遗传分析，运用毛细管电泳检测可准确定位多态位点，一定程度上弥补了传统聚丙烯酰胺凝胶电泳的缺点。 |
| 关键词: 白杨派 毛细管电泳 限制性位点扩增多态性 指纹图谱 遗传分析 |
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| 基金项目:“十三五”国家重点研发计划项目（SQ2016YFNC030041） |
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| Fingerprinting and genetic analysis of ten poplar clones based on RSAP markers |
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CHENG Weizhe,FAN Junfeng,GAO Jianshe,et al
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| Abstract: |
| 【Objective】This study evaluated the identification effect of RSAP markers on improved clones of poplar,and provided a basis for the protection of poplar germplasm resources and indentification.【Method Based on 10 excellent clones of Populus (Qinbaiyang 1,Qinbaiyang 3,Qinbaiyang 5,84K,I-101,Xinjiangyang,Maobaiyang 30,07-17-18,07-23-23 and 07-30-11),6 pairs of primers were selected from 36 pairs of primer combinations composed of 9 RSAP primers.Capillary electrophoresis and POPGENE 32 software were used to detect and analyze polymorphisms of amplified products and calculate related indicators of genetic diversity.Primers with high polymorphism and ability to distinguish all clones were selected to construct fingerprints of 10 clones of Sect.Leuce.NTSYS 2.1 software was used to calculate genetic similarity coefficient among clones,and UPGMA was used for cluster analysis.【Result】A total of 133 polymorphic loci were amplified from 10 clones by 6 primer combinations,with an average of 22.17 per primer pair.The average Shannon information index was 0.326 and average Nei’s gene diversity index was 0.488,indicating that there was high genetic diversity and rich genetic variation among clones.All of the 10 clones could be identified by using 6 polymorphic loci of 2 primer combinations Rs2/Rs4 and Rs2/Rs7.The genetic similarity coefficients of the 10 clones ranged from 0.400 to 0.817 with average of 0.594.Cluster analysis showed that 10 clones of Sect.Leuce were divided into 2 groups when the genetic similarity coefficient was 0.510,Xinjiangyang and Maobaiyang 30 in one group and the rest in another group, which was roughly consistent with actual genetic relationship.【Conclusion】RSAP molecular markers can be used effectively for poplar germplasm identification and genetic analysis.Capillary electrophoresis can accurately locate polymorphic loci and offset the shortcomings of traditional polyacrylamide gel electrophoresis. |
| Key words: Sect.Leuce capillary electrophoresis restriction site amplified polymorphism (RSAP) fingerprinting genetic analysis |
