文献类型：期刊文献

中文题名：基于桤木属转录组测序的SSR分子标记的开发

英文题名：Development of SSR Molecular Markers Based on Transcriptome Sequences of Alnus

作者：饶龙兵[1] 杨汉波[1] 郭洪英[2] 段红平[3] 陈益泰[1]

第一作者：饶龙兵

机构：[1]中国林业科学研究院亚热带林业研究所;[2]四川省林业科学研究院;[3]云南农业大学资源与环境学院

年份：2016

卷号：29

期号：6

起止页码：875-882

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金："十二五"国家科技支撑计划项目桤木育种专题"抗逆生态树种桤木新品种选育技术研究"(2012BAD01B0604)

语种：中文

中文关键词：桤木属;转录组;SSR;引物

外文关键词：Alnus; transcriptome; SSR; primer

分类号：S718.46

摘要：[目的]基于转录组数据开发适用于桤木属树种的SSR标记,揭示其在转录组序列中的分布类型及特征,为桤木属树种分子标记辅助育种提供有利工具。[方法]利用Micro SAtellite(MISA)软件对所有转录组序列进行SSRs搜索,并对SSR位点的数量、分布特征进行统计分析。设计100对SSR引物,采用琼脂糖凝胶电泳和毛细管电泳分离检测方法对3种不同倍性桤木属植物(12份材料)进行遗传多样性检测,确定引物多态性及通用性。[结果]85 769条Unigenes序列中发现8 678个SSR位点,分布在8 298条Unigenes中发生频率为9.67%,转录组序列中平均每14.04 kb长度就有一个SSR位点分布。其中,二核苷酸重复类型数量最多,占65.87%。根据转录组Unigenes序列,利用Primer 3软件共设计出4 531对符合要求的引物,挑选出的100对SSR引物中,获得18对多态性高、稳定性好的SSR引物。[结论]本研究可扩增出多态性位点的引物重复单元以二、三核苷酸重复为主。基于桤木属转录组序列的SSR标记开发是可行的,开发的引物为桤木属遗传多样性分析、分子育种、遗传图谱构建和功能基因的挖掘提供了丰富的候选分子标记。
[Objective]To develop SSR markers of gene transcriptional areas from Alnus based on the results of transcriptome sequencing. [Methods]Distribution patterns of the markers in the transcriptome sequences and their characteristics were analyzed,in order to provide more powerful tools for molecular marker-assisted breeding Alnus.The SSR locus from transcriptome sequences were searched by Micro SAtellite（ MISA）,and statistical analyses were conducted for the amount,distribution and characteristics of SSR loci. 100 pairs of SSR primers were designed and synthesized. Agarose electrophoresis was used for initial check and capillary electrophoresis for separation and detection of the polymorphism of the primers. [Results]A total of 8 298 Unigenes containing 8 678 SSR locus were searched from 85 769 Unigenes by Micro SAtellite（ MISA） sequence analysis software,accounting for 9. 67% of the transcriptome sequences,with an average of one SSR per 14. 04 kb. The dinucleotide repeat is the most abundant repeat type,accounting for 65. 87% of the total number of SSRs. Additionally,a small amount of GC repeats was found. A total of 4 531 pairs of primers were designed by Primer 3 software according to the Unigene sequences oftranscriptional. 18 pairs of primers produced bands with expected sizes were selected from 100 pairs of primers.[Conclusion]The amplified primers of the polymorphism loci were mainly dinucleotide and trinucleotide repeats.This study has an important value to develop SSR molecular markers based on the transcriptome sequencing analysis. This study is also important for analyzing genetic diversity,marker assisted selection,genetic linkage mapping and functional gene mining of Alnus by using SSR molecular markers.