Chen Zhi, Jin Chen, Wang Xiaoyun, Deng Yaqiong, Tian Xiaodan, Li Xunxun, Zhang Qinxi, Zeng Yanli, Liao Jiawei, Zhang Ling
School of Pharmacy, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi 330004, China.
Key Laboratory of Modern Preparation of TCM, Ministry of Education, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi 330004, China.
J AOAC Int. 2022 Dec 22;106(1):146-155. doi: 10.1093/jaoacint/qsac097.
Callerya reticulata (Bentham) Schot, Callerya dielsiana (Harms) P.K. Loc ex Z. Wei & Pedley, Callerya nitida var. hirsutissima (Z. Wei) X.Y. Zhu, and Callerya nitida (Bentham) R. Geesink, which belongs to the Leguminosae family, are important medicinal plants in China. The genus Callerya includes 26 species, 18 species are distributed in China, and the vine stems of some species are used as traditional medicinal herbs because they have important pharmacological activity. Due to the high similarity of appearance, it is difficult to identify them in the market by appearance alone. Therefore, circulating of Callerya-related materia medica on the market is confusing, sometimes even leading to drug safety problems. It is urgent to develop molecular methods for their identification.
To sequence and analyze the complete chloroplast (cp) genomes of C. reticulata, C. dielsiana, C. nitida var. hirsutissima, and C. nitida and to analyze their cp genome differences as a basis for seeking easier DNA barcoding for their identification.
After using Illumina high-throughput sequencing and nanopore sequencing to obtain the genome data, some bioinformatics software was used to assembly and analyze the molecular structure of cp genomes.
The complete cp genomes of the four species were circular molecules, which ranged from 130 435 to 132 546 bp, and GC contents ranged from 33.89% to 34.89%. Each of them includes a large single-copy region, a small single-copy region, and without large inverted repeat regions.
These results suggested that highly variable regions of the four cp genomes would provide useful plastid markers, which could be used as a potential genomic resource to resolve phylogenetic questions and provide a reference for mining specific DNA barcodes of these species.
Our study provided highly effective molecular markers for subsequent phylogenetic analysis, species identification, and biogeographic analysis of Callerya.
豆科的网脉鸡血藤（Callerya reticulata (Bentham) Schot）、中南鸡血藤（Callerya dielsiana (Harms) P.K. Loc ex Z. Wei & Pedley）、毛瓣亮叶鸡血藤（Callerya nitida var. hirsutissima (Z. Wei) X.Y. Zhu）和亮叶鸡血藤（Callerya nitida (Bentham) R. Geesink）是中国重要的药用植物。鸡血藤属包含26个物种，其中18个物种分布于中国，一些物种的藤茎因其具有重要的药理活性而被用作传统草药。由于外观高度相似，仅通过外观在市场上很难对它们进行鉴别。因此，市场上鸡血藤相关药材的流通较为混乱，有时甚至会导致用药安全问题。迫切需要开发其分子鉴别方法。
对网脉鸡血藤、中南鸡血藤、毛瓣亮叶鸡血藤和亮叶鸡血藤的叶绿体（cp）全基因组进行测序和分析，分析其cp基因组差异，为寻找更简便的DNA条形码进行鉴别提供依据。
利用Illumina高通量测序和纳米孔测序获得基因组数据后，使用一些生物信息学软件对cp基因组的分子结构进行组装和分析。
这4个物种的cp全基因组均为环状分子，长度在130435至132546 bp之间，GC含量在33.89%至34.89%之间。每个cp基因组均包括一个大单拷贝区、一个小单拷贝区，无大的反向重复区。
这些结果表明，这4个cp基因组的高变区将提供有用的质体标记，可以作为一种潜在的基因组资源来解决系统发育问题，并为挖掘这些物种的特异性DNA条形码提供参考。
本研究为后续鸡血藤属的系统发育分析、物种鉴定和生物地理学分析提供了高效的分子标记。
