Position: Home > Articles > Development of SSR Markers of Fanjingshan Paris polyphylla Smith var. chinensis Based on High Throughput RNA-seq
Molecular Plant Breeding
2019
(18)
6059-6065
高通量RNA-seq技术开发梵净山重楼SSR标记
作 者：
王海明;阮成江;梁玖华;韩平;闫蕊;董旭;刘祾悦;帅小春;刘贺贺
单 位：
大连民族大学资源植物研究所生物技术与资源利用教育部重点实验室
关键词：
重楼(Paris polyphylla Smith var. chinensis);RNA-seq;SSR标记
摘 要：
梵净山重楼(Paris polyphylla Smith var. chinensis)是中国重要的中草药之一。本研究利用MISA软件从重楼块根高通量转录组数据库的38 802条Unigenes中共搜索挖掘出6 333个SSR标记。66.76%的二核苷酸重复是这些SSR标记的主要重复类型,然后是18.77%的三核苷酸重复、11.50%的单核酸苷酸重复及2.97%的四核苷酸、五核苷酸和六核苷酸重复。在所有被功能注释的Unigenes中,481条Unigenes涉及调控重楼皂苷、黄酮、种子萌发和种子生长发育,从这些Unigenes共搜索挖掘到90个SSR标记。通过对这90个SSR标记的有效性验证,获得具有清晰扩增条带的SSR标记20对。以选取的14份重楼种质为材料,对扩增出清晰条带的20对SSR标记进行多态性检测后,获得16对具有多态性的SSR标记引物。引物多态性信息含量指数(polymorphism information content, PIC)介于0.067～0.561,平均值为0.293;14个重楼种质聚类在两个不同类群。本研究结果可为梵净山重楼种质资源鉴定、遗传多样性分析和分子标记辅助选择育种提供SSR标记。
译 名：
Development of SSR Markers of Fanjingshan Paris polyphylla Smith var. chinensis Based on High Throughput RNA-seq
作 者：
Wang Haiming;Ruan Chengjiang;Liang Jiuhua;Han Ping;Yan Rui;Dong Xu;Liu Lingyue;Shuai Xiaochun;Liu Hehe;Key Laboratory of Biotechnology and Bioresources Utilization,Ministry of Education,Institute of Plant Resources,Dalian Minzu University;Institute of Biological Resources of Fanjingshan,Tongren Academy of Sciences;
关键词：
Paris polyphylla Smith var.chinensis;;RNA-seq;;SSR marker
摘 要：
Paris polyphylla Smith var. chinensis in Fanjingshan is one of an important Chinese medicinal herbs. A total of 6 333 SSR markers were discovered in the 38 802 unigenes using MISA software, which was from high throughput RNA-seq data of root tubers of P. polyphylla Smith var. chinensis in this study. 66.76% of dinucleotide repeats were the major repeat type of these SSR markers, followed by 18.77% of trinucleotide repeats, 11.50% of single nucleotide repeats, and 2.97% of tetranucleotides, pentanucleotides and Hexanucleotide repeat. In the function annotation of 38 802 unigenes, 481 unigenes were annotated the functions of regulating biosynthesis of saponins and flavonoids, seed germination and growth of root tuber. 90 SSR markers were mined in these targeted unigenes, 20 pairs of these markers were found with clear targeted PCR production based on the preliminary verification of the effectiveness of these SSR markers. 14 accessions of P. polyphylla Smith var. chinensis were used as materials to screen polymorphic SSR markers, and the results showed that 16 of 20 pairs of primers were polymorphic. The polymorphism information content(PIC) ranged from 0.067 to 0.561 with an average of 0.293,and 14 accessions were clustered into two groups. The results of this study could provide SSR markers for the identification of germplasm resources, genetic diversity analysis and molecular marker-assisted selection breeding of Fanjingshan P. polyphylla Smith var. chinensis.
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