Comparative Chloroplast Genome Analysis of Rhubarb Botanical Origins and the Development of Specific Identification Markers.
Comparative Chloroplast Genome Analysis of Rhubarb Botanical Origins and the Development of Specific Identification Markers.
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大黄植物起源的比较叶绿体基因组分析和特异性识别标记的开发。
DOI：
10.3390/molecules23112811
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发表时间：
2018-10-30
期刊：
影响因子：
--
通讯作者：
Wang B
中科院分区：
文献类型：
--
作者：
Zhou Y;Nie J;Xiao L;Hu Z;Wang B
大黄是传统中药大黄的重要成分。然而，这个通用名称是指三种不同的植物物种，具有不同的药理作用。为了便于这三个物种的遗传鉴定，更准确地应用于中药，我们在这里希望提供叶绿体序列的特异性识别位点，易于扩增。因此，我们对所有三个物种的叶绿体基因组进行了完整的测序，然后筛选了描述这三个物种的合适序列。3个叶绿体基因组的长度为161，053 ~ 161，541 bp，共编码131个基因，包括31个tRNA、8个rRNA和92个蛋白质编码序列。简单重复序列分析表明，大黄属3个种间存在差异，系统发育分析表明，叶绿体基因组可以作为区分大黄属3个种的超级条形码，非编码区的变异高于蛋白质编码区，单拷贝区的变异高于反向重复区。设计了21对特异性引物，并通过实验确定了8个特异性识别位点，可作为特异性DNA条形码用于基于高度可变区的3个物种的鉴定。本研究为大黄属药用植物的精确筛选提供了分子基础，也为下一步大黄属近缘种的比较研究和这些重要药用种的正确鉴定奠定了基础。
Rhubarb is an important ingredient in traditional Chinese medicine known as Rhei radix et rhizome. However, this common name refers to three different botanical species with different pharmacological effects. To facilitate the genetic identification of these three species for their more precise application in Chinese medicine we here want to provide chloroplast sequences with specific identification sites that are easy to amplify. We therefore sequenced the complete chloroplast genomes of all three species and then screened those for suitable sequences describing the three species. The length of the three chloroplast genomes ranged from 161,053 bp to 161,541 bp, with a total of 131 encoded genes including 31 tRNA, eight rRNA and 92 protein-coding sequences. The simple repeat sequence analysis indicated the differences existed in these species, phylogenetic analyses showed the chloroplast genome can be used as an ultra-barcode to distinguish the three botanical species of rhubarb, the variation of the non-coding regions is higher than that of the protein coding regions, and the variations in single-copy region are higher than that in inverted repeat. Twenty-one specific primer pairs were designed and eight specific identification sites were experimentally confirmed that can be used as special DNA barcodes for the identification of the three species based on the highly variable regions. This study provides a molecular basis for precise medicinal plant selection, and supplies the groundwork for the next investigation of the closely related Rheum species comparing and correctly identification on these important medicinal species.
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