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基于DNA条形码技术鉴定僵蚕

[Identification of Bombyx Batryticatus based on DNA barcoding technology].

作者信息

Jia Jing, Shi Lin-chun, Yao Hui, Song Jing-yuan, Chen Shi-lin

出版信息

Yao Xue Xue Bao. 2016 Nov;51(11):1784-90.

PMID:29908528
Abstract

To identify the commercial medicinal materials of Bombyx Batryticatus, two-dimensional DNA barcode was used to construct the "Internet Plus" identification system for Chinese medicine, which should benefit the cross-platform communication of DNA barcode information. Bombyx Batryticatus contained Bombyx mori Linnaeus and Beauveria bassiana (Bals.) Vuillant. Both COI and ITS sequences were obtained via PCR amplification for total genomic DNA extracted from raw materials using the animal genomic DNA kit, while only ITS but no COI sequences was obtained when using the plant genomic DNA kit. The ITS sequences obtained using the animal genomic DNA kit were consistent with those using plant genomic DNA kit. The medicinal materials yielded COI sequences and identified as B. mori. According to analysis of ITS sequences, the main species of the medicinal materials were identified as B. bassiana and few were identified as other fungi. NJ trees analysis based on ITS sequences suggests that it can be easily distinguished from other fungi. Our results showed that total genomic DNA of B. mori and B. bassiana was extracted simultaneously using the animal genomic DNA kit, which could effectively solve the problem in species identification of animal and fungi mixture materials. COI and ITS regions as DNA barcodes can stably and accurately identify Bombyx Batryticatus. The "Internet Plus" two-dimensional DNA barcode system will promote the standardization and normalization of Chinese medicinal materials market.

摘要

为鉴定僵蚕的商品药材,采用二维DNA条形码构建中药“互联网 +”鉴定系统,这将有利于DNA条形码信息的跨平台交流。僵蚕包含家蚕(Bombyx mori Linnaeus)和白僵菌(Beauveria bassiana (Bals.) Vuillant)。使用动物基因组DNA提取试剂盒从原材料中提取总基因组DNA,通过PCR扩增获得COI和ITS序列,而使用植物基因组DNA提取试剂盒时仅获得ITS序列,未获得COI序列。使用动物基因组DNA提取试剂盒获得的ITS序列与使用植物基因组DNA提取试剂盒获得的一致。药材产生COI序列并鉴定为家蚕。根据ITS序列分析,药材的主要种类鉴定为白僵菌,少数鉴定为其他真菌。基于ITS序列的NJ树分析表明,它可以很容易地与其他真菌区分开来。我们的结果表明,使用动物基因组DNA提取试剂盒可同时提取家蚕和白僵菌的总基因组DNA,这能有效解决动物和真菌混合材料物种鉴定的问题。COI和ITS区域作为DNA条形码能够稳定、准确地鉴定僵蚕。“互联网 +”二维DNA条形码系统将促进中药材市场的标准化和规范化。

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