Li Qing, Li Biao, Guo Shun-Xing
Institute of medicinal Plant Development, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100193, China.
Zhongguo Zhong Yao Za Zhi. 2017 Jan;42(1):63-69. doi: 10.19540/j.cnki.cjcmm.20161222.044.
SSR is one of the most important molecular markers used in molecular identification and genetic diversity research of Dendrobium nobile. In order to enrich the library of SSR and establish a method for rapid identification of D. nobile, the SSR information was analyzed in the transcriptome of D. nobile. A total of 32 709 SSRs were obtained from the transcriptome of D. nobile, distributed in 26 742 unigenes with the distribution frequency of 12.90%. SSR loci occurred every 3 748 bp. Mono-nucleotide repeat was the main type, account for as much as 72.18% of all SSRs, followed by di-nucleotide (15.97%) and tri-nucleotide (11.19%). Among all repeat types, A/T was the predominant one followed by AG/CT. Finally a total of 62 157 primer pairs were designed for marker development. Randomly 20 pairs of primers were selected for PCR amplification, 17 amplified on clear and reproducible bands, the amplification rate was 85.0%.Thirteen pairs were polymorphic among the 3 Dendrobium plants. The results indicated that the unigenes generated from transcriptome sequencing in D. nobile can be used as effective source to develop SSR markers. The SSR loci in the transcriptome of D. nobile have the characteristics of type riches, high density and high potential of polymorphism, and these characteristics might applied in the study of molecular identification, genetic diversity and marker-assisted breeding of D. nobile and its closely related species.
简单重复序列(SSR)是用于铁皮石斛分子鉴定和遗传多样性研究的最重要分子标记之一。为了丰富SSR文库并建立铁皮石斛快速鉴定方法,对铁皮石斛转录组中的SSR信息进行了分析。从铁皮石斛转录组中总共获得了32709个SSR,分布在26742个单基因中,分布频率为12.90%。SSR位点每3748 bp出现一次。单核苷酸重复是主要类型,占所有SSR的72.18%,其次是二核苷酸(15.97%)和三核苷酸(11.19%)。在所有重复类型中,A/T是主要的,其次是AG/CT。最后共设计了62157对引物用于标记开发。随机选择20对引物进行PCR扩增,17对扩增出清晰且可重复的条带,扩增率为85.0%。在3种石斛属植物中,有13对具有多态性。结果表明,铁皮石斛转录组测序产生的单基因可作为开发SSR标记的有效来源。铁皮石斛转录组中的SSR位点具有类型丰富、密度高和多态性潜力大的特点,这些特点可应用于铁皮石斛及其近缘种的分子鉴定、遗传多样性和标记辅助育种研究。