Subtropical Plant Science >

2020 , Vol. 49 >Issue 06: 411 - 419

DOI: https://doi.org/10.3969/j.issn.1009-7791.2020.06.001

Plant biochemistry and molecular biology

Screening of DNA Barcoding for Medicinal Dendrobium in Yunnan

  • XU Su-su ,
  • GAO Jing ,
  • CHEN Jun-wen ,
  • LIU Xiang-yu ,
  • ZHANG Jing-li
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  • (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 date: 2020-10-30

  Revised date: 2020-12-12

  Online published: 2020-12-30

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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

Cite this article

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 . DOI: 10.3969/j.issn.1009-7791.2020.06.001

References

[1] 宋广青,刘新民,王琼,刘靖. 石斛药理作用研究进展[J]. 中草药, 2014,45(17): 2576–2580. [2] He L, Su Q, Bai L, Li M F, Guo L. Recent research progress on natural small molecule bibenzyls and its derivatives in Dendrobium species[J]. European Journal of Medicinal Chemistry, 2020,204: 112530. [3] Nie X Q, Chen Y, Li W, Lu Y L. Anti-aging properties of Dendrobium nobile Lindl.: From molecular mechanisms to potential treatments[J]. Journal of Ethnopharmacology, 2020,257: 112839. [4] Yoo S R, Jeong S J, Lee N R, Shin H K, Seo C S. Simultaneous determination and anti-inflammatory effects of four phenolic compounds in Dendrobii Herba[J]. Natural Product Research, 2017,31(24): 2923–2926. [5] 贾元,胡利娟,冯群,田忠静,余雨,翁庆北,朱斌. 基于ISSR标记的西南地区石斛资源亲缘关系分析[J]. 贵州农业科学, 2019,47(12): 8–12. [6] 中国科学院中国植物志编辑委员会. 中国植物志(第19卷)[M]. 北京: 科学出版社, 1999: 67–68. [7] 中国科学院昆明植物研究所. 云南植物志(第14卷)[M]. 北京: 科学出版社, 2003. [8] 白音,包英华,金家兴,阎玉凝,王文全. 我国药用石斛资源调查研究[J] .中草药, 2006,37 (9): 4–6. [9] 姚能昌. 浅述云南石斛资源现状及开发利用技术[J]. 林业调查规划, 2004,29 (4): 80–82. [10] 云南省药材公司. 云南中药资源名录[M]. 北京: 科学出版社, 1993: 669–672. [11] 贾敏如,李星炜. 中国民族药志要[M]. 北京: 中国医药科技出版社, 2005: 217–218. [12] 黄海,李劲松,符岸军,严海. 石斛属植物DNA条形码序列的筛选[J]. 热带作物学报, 2010,31(10): 1769–1777. [13] 陈士林,郭宝林,张贵君,严铸云,罗光明,孙素琴,吴和珍,黄林芳,庞晓慧,陈建波. 中药鉴定学新技术新方法研究进展[J]. 中国中药杂志, 2012,37(8): 1043–1055. [14] Li D Z, Liu J Q, Chen Z D, Wang H, Ge X J, Zhou S L, Gao L M, Fu C X, Chen S L. Plant DNA barcoding in China[J]. Journal of Systematics and Evolution.2011,49(3): 165–168. [15] 程芳婷,李忠虎,刘春艳,原超,李雪童,刘占林. 地黄属植物的DNA条形码研究[J]. 植物科学学报, 2015,33 (1): 25–32. [16] Kress W J, Wurdack K J, Zimmer E A, Weigt L A, Janzen D H. Use of DNA barcodes to identify flowering plants[J]. Proceedings of the National Academy of Sciencesof the United States of America, 2005,102: 869–8374. [17] Chen S L, Yao H, Han J P, Liu C, Song J Y, Shi L C, Zhu Y J, Ma X Y, Gao T, Pang X H, Luo K, Li Y, Li X W, Jia X C, Lin Y L, Leon C. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS ONE, 2010,5: e8613. [18] Hollingsworth P M. Refining the DNA barcode for land plants[J]. Proceedings of the National Academy of Sciences of the United States of America, 2011,108(49): 19451–2. [19] 陈士林,姚辉,韩建萍,辛天怡,庞晓慧,石林春,罗焜,宋经元,侯典云,石上梅,钱忠直. 中药材DNA条形码分子鉴定指导原则[J]. 中国中药杂志, 2013,38(2): 141–148. [20] 国家药典委员会. 中国药典(一部)[M]. 北京: 中国医药科技出版社, 2015. [21] Shi L C, Zhang J, Han J P, Song J Y, Yao H, Zhu YJ, Li J C, Wang Z Z, Xiao W, Lin Y L, Xie C X, Qian Z Z, Chen S L. Testing the potential of proposed DNA barcodes for species identification of Zingiberaceae[J]. Journal of Systematics and Evolution, 2011,49(3): 261–266. [22] Ran J H, Wang P P, Zhao H J, Xiao Q W. A test of seven candidate barcode regions from the plastome in Picea (Pinaceae)[J]. Journal of Integrative Plant Biology, 2010,52(12): 1109–1126. [23] Zhang J, Chen M, Dong X Y, Lin R Z, Fan J H, Chen Z D. Evaluation of four commonly used DNA barcoding loci for Chinese medicinal plants of the family Schisandraceae[J]. PLoS One, 2015,10(5): e125574. [24] Zhang C Y, Wang F Y, Yan H F, Hao G, Hu C M, Ge X J. Testing DNA barcoding in closely related groups of Lysimachia L. (Myrsinaceae) [J]. Molecular EcologyResources., 2012,12: 98–108. [25] 刘枫,赵群,戴军,陈存武,陈乃富,韩邦兴. DNA分子标记技术在石斛属鉴别中的应用进展[J]. 皖西学院学报, 2017(2): 14–18,36. [26] 叶子,卢叶,王峥涛,徐红,胡之璧. 基于ITS序列鉴别石斛类药材[J]. 中国中药杂志, 2014,39(20): 3928–3935. [27] Lau D T, Shaw P C, Wang J, Bu P. Authentication of medicinal Dendrobium Species by the internal transcribed spacer of ribosomal DNA [J]. Planta Med, 2001,67(5): 456–460. [28] Yao H, Song J Y, Ma X Y, Liu C, Li Y, Xu H X, Han J P, Duan L S, Chen S L. Identification of Dendrobium Species by a candidate DNA barcode sequence: The chloroplast psbA-trnH intergenic region [J]. Planta Med, 2009,75(6): 667–669. [29] 李金璐,王硕,于婧,王玲,周世良. 一种改良的植物DNA提取方法[J]. 植物学报, 2013,48(1): 72–78. [30] Keller A, Schleicher T, Schultz J, Muller T, Dandekar T, Wolf M. 5.8S-28S rRNA interaction and HMM-based ITS2 annotation[J].Gene, 2009,430(1–2): 0–57. [31] Meyer P, Paulay G. DNA barcoding: Error rates based on comprehensive sampling[J]. PLoS Biology, 2005,3(12): e422 [32] 刘依丽,冯尚国,何仁锋,赵妙玉,吴慧,崔艺庆,陈倩,王慧中. 基于ITS2条形码对铁皮石斛及其混伪品分子鉴定的初步研究[J]. 杭州师范大学学报(自然科学版), 2014(1): 35–41. [33] 邵世光,韩丽,马艳红,沈洁,张伟超,丁小余. 枫斗类石斛cpDNA psbA–trnH的序列分析与鉴别[J]. 药学学报, 2009(10): 1173–1178. [34] 滕艳芬,吴晓俊,徐红,王峥涛,余国奠,徐珞珊. 石斛及其常见混淆品的matK基因序列比较[J]. 中国药科大学学报, 2002,33(4): 280–283. [35] 张爱丽,高亚芳,黎太米,谭文红,钱子刚. DNA条形码在主要滇产石斛鉴别中的应用[J]. 中国民族民间医药, 2017,26(2): 11–16. [36] Li D Z, Gao L M, Li H T, Wang H, Ge X J, Liu J Q, Chen D Z, Zhou S L, Chen S L, Yang J B. Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants[J]. Proceedings of the National Academy of Sciences, 2011,108(49): 19641–19646. [37] Kress W J, Wurdack K J, Zimmer E A, Weigt L A, Janzen D H. Use of DNA barcodes to identify flowering plants[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005,102(23): 8369.
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