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Characterization of Global Transcriptome Using Illumina Paired-End Sequencing and Development of EST-SSR Markers in Two Species of Gynostemma (Cucurbitaceae)

机译:在两个物种的绞股蓝(葫芦科)中使用照度配对末端测序对全球转录组进行鉴定和EST-SSR标记的开发。

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摘要

Gynostemma pentaphyllum is an important medicinal herb of the Cucurbitaceae family, but limited genomic data have hindered genetic studies. In this study, transcriptomes of two closely-related Gynostemma species, Gynostemma cardiospermum and G. pentaphyllum, were sequenced using Illumina paired-end sequencing technology. A total of 71,607 nonredundant unigenes were assembled. Of these unigenes, 60.45% (43,288) were annotated based on sequence similarity search with known proteins. A total of 11,059 unigenes were identified in the Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG) database. A total of 3891 simple sequence repeats (SSRs) were detected in 3526 nonredundant unigenes, 2596 primer pairs were designed and 360 of them were randomly selected for validation. Of these, 268 primer pairs yielded clear products among six G. pentaphyllum samples. Thirty polymorphic SSR markers were used to test polymorphism and transferability in Gynostemma. Finally, 15 SSR makers that amplified in all 12 Gynostemma species were used to assess genetic diversity. Our results generated a comprehensive sequence resource for Gynostemma research.
机译:绞股蓝(Gynostemma pentaphyllum)是葫芦科的重要草药,但是有限的基因组数据阻碍了遗传研究。在这项研究中,使用Illumina的双末端测序技术对两个紧密相关的绞股蓝物种Gynostemma cardiospermum和G. pentaphyllum的转录组进行了测序。总计71,607个非冗余单基因。在这些单基因中,基于与已知蛋白质的序列相似性搜索进行注释的占60.45%(43,288)。在《京都市基因与基因组途径百科全书》(KEGG)数据库中总共鉴定出11,059个单基因。在3526个非冗余单基因中共检测到3891个简单序列重复(SSR),设计了2596个引物对,并随机选择其中360个进行验证。其中,有268对引物在六个G.pentaphyllum样品中产生了透明产物。三十个多态性SSR标记用于测试绞股蓝的多态性和可转移性。最后,使用在所有12个绞股蓝物种中扩增的15个SSR标记物来评估遗传多样性。我们的结果为绞股蓝的研究提供了一个全面的序列资源。

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