芒果EST-SSR位点分析及引物开发

Analysis of EST-SSR Loci and Primers Development in Mango(Mangifera indica)

本研究通过芒果转录组数据开发EST-SSR标记,以期为芒果的分子辅助育种和不同品种的资源鉴定提供有效的检测手段。利用MISA软件分析芒果叶片和果实两个组织转录组结果中得到的SSR信息,经PCR验证扩增产物,分析不同品种间的亲缘关系。对芒果组装的99328个Unigene中,其中含有SSR的序列Unigene数目为4877个,发生频率,即含SSR的Unigene数目/总Unigene数目(4877/99328)是4.91%。芒果转录组中查到SSR位点6405个。从中随机筛选31对引物,经PCR扩增后进行6%非变性聚丙烯酰胺凝胶电泳,并选出8对引物扩增具有多态性。利用DPS数据处理软件对得到的条带数据化并进行非加权组平均法UPGMA聚类分析,构建系统发生树得到8个芒果品种间的遗传距离和亲缘关系。芒果转录组测序EST-SSR分析能够为开发EST-SSR标记提供理论依据,此标记可为芒果遗传多样性分析、遗传亲缘关系及图谱构建提供研究基础。

In this study,EST-SSR markers were developed from Mango transcriptome data,in order to provide effective means for molecular assisted breeding and identification of different varieties of mango.The SSR information obtained from transcriptome results of mango was analysed by MISA.The amplified products were verified by PCR to analyse the genetic relationship among eight different varieties.Among 99328 Unigenes assembled in mango transcriptome,the number of sequences containing SSR was 4877.The occurrence frequency,the number of Unigenes containing SSR/the total number of Unigenes(4877/99328),was 4.91%.The number of SSR loci was 6405 based on the results of mango transcriptomes.A total of 31 pairs of primers were randomly selected.After amplified,PCR products were conducted electrophoresis in 6%non-denatured polyacrylamide gel.A total of eight pairs of primers showed polymorphism.These data were analysed by DPS data processing software.The genetic distance and genetic relationship of eight mango cultivars were obtained by constructed a phylogenetic tree using UPGMA clustering analysis.These results can provide theoretical basis for the development of EST-SSR markers,which can provide research basis for genetic diversity analysis,genetic relationship and map construction.