云南常见药用石斛DNA条形码筛选

doi:10.3969/j.issn.1009-7791.2020.06.001

亚热带植物科学 ›› 2020, Vol. 49 ›› Issue (06): 411-419.DOI: 10.3969/j.issn.1009-7791.2020.06.001

• 植物生化与分子生物学 • 下一篇

云南常见药用石斛DNA条形码筛选

徐素素^1,2,3，高静^1,2,3，陈军文^2,3,4，刘祥宇^1,2,3，张敬丽^1,2,3*

(1.云南农业大学园林园艺学院，云南昆明 650201；2.云南农业大学西南中药材种质创新与利用国家地方联合工程研究中心，云南昆明 650201；3.云南农业大学云南省药用植物生物学重点实验室，云南昆明650201；4.云南农业大学农学与生物技术学院，云南昆明 650201)

收稿日期:2020-10-30 修回日期:2020-12-12 出版日期:2020-12-30 发布日期:2020-12-30
通讯作者: 张敬丽
基金资助:
云南省重大科技专项(2017ZF003)；国家重点研发计划(2017YFC1702205)；云南省教育厅科学研究基金项目(2021Y184)

Screening of DNA Barcoding for Medicinal Dendrobium in Yunnan

XU Su-su^1,2,3, GAO Jing^1,2,3, CHEN Jun-wen^2,3,4, LIU Xiang-yu^1,2,3, ZHANG Jing-li^1,2,3*

(1.College of Landscape and Horticulture, Yunnan Agricultural University, Kunming 650201, Yunnan China; 2.National & Local Joint Engineering Research Center on Germplasm Innovation & Utilization of Chinese Medicinal Materials in Southwestern China, Yunnan Agricultural University, Kunming 650201, Yunnan China; 3.The Key Laboratory of Medicinal Plant Biology of Yunnan Province, Yunnan Agricultural University, Kunming 650201, Yunnan China; 4.College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, Yunnan China)

Received:2020-10-30 Revised:2020-12-12 Online:2020-12-30 Published:2020-12-30
Contact: ZHANG Jing-li

摘要/Abstract

摘要： 选择云南省内常见的12种药用石斛，采用分子生物学鉴定技术筛选其适用的DNA条形码序列。以12种药用石斛共36个样品为材料，提取样品总DNA，对核基因片段ITS和ITS2、叶绿体基因片段psbA-trnH和matK序列进行扩增、测序，结合GenBank下载部分石斛序列；利用生物信息学软件进行序列分析和系统发育分析。结果表明，4条序列扩增和测序成功率均为100%；4条序列没有明显的Barcoding Gap，但ITS序列与ITS2序列的种内和种间重叠部分较少，有偏向两端的趋势；系统发育树显示，ITS和psbA-trnH序列能成功区分12种云南常见的药用石斛，ITS2序列未能区分长距石斛(Dendrobium longicornu)和矮石斛(D. bellatulum)，matK序列仅区分6种石斛。建议以ITS和psbA-trnH序列作为云南药用石斛鉴定序列，为药用石斛的种源鉴定提供理论依据。

关键词: 药用石斛, 云南, DNA条形码, 系统发育分析, 物种鉴定, 分子鉴定

Abstract: Twelve common medicinal Dendrobium species in Yunnan province were selected, and molecular biology identification techniques were used to screen DNA barcoding sequences suitable for Yunnan medicinal Dendrobium. A total of 36 samples of 12 kinds of medicinal Dendrobium (Dendrobium devonianum, D. chrysotoxum, D. falconeri, etc.) were used as materials to extract the total DNA of the samples, and the nuclear gene fragments ITS and ITS2, the chloroplast gene fragment psbA-trnH and matK sequences were amplified and sequenced, part of the Dendrobium sequence was downloaded in GenBank; Sequence analysis and phylogenetic analysis were used by bioinformatics software. The results showed that the success rates of amplification and sequencing of the 4 sequences were 100% and all of them had no obvious Barcoding Gap, but there was a small amount of overlap between the ITS sequence and the ITS2 sequence within and between species, which tended to be biased towards both ends. The phylogenetic tree showed that the ITS and psbA-trnH sequences could successfully distinguish the 12 medicinal Dendrobiums in this study, the ITS2 sequence failed to distinguish D. longicornu and D. bellatulum. The matK sequence only distinguished 6 species of Dendrobium. It is suggested that ITS and psbA-trnH sequences should be regarded as the identification sequence of Yunnan medicinal Dendrobium to provide theoretical basis for the provenance identification of medicinal Dendrobium.

Key words: medicinal Dendrobium, Yunnan, DNA barcoding, phylogenetic analysis, species identification, molecular identification

中图分类号:

S567

徐素素，高静，陈军文，刘祥宇，张敬丽. 云南常见药用石斛DNA条形码筛选[J]. 亚热带植物科学, 2020, 49(06): 411-419.

XU Su-su, GAO Jing, CHEN Jun-wen, LIU Xiang-yu, ZHANG Jing-li . Screening of DNA Barcoding for Medicinal Dendrobium in Yunnan[J]. Subtropical Plant Science, 2020, 49(06): 411-419.

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云南常见药用石斛DNA条形码筛选

Screening of DNA Barcoding for Medicinal Dendrobium in Yunnan

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