摘要
目的:基于性状鉴别和DNA条形码联用技术建立太平洋海马药材(Hippocampus ingens)的生药学鉴定特征,为其后续的应用开发奠定基础。方法:利用传统性状鉴别方法研究太平洋海马的典型性状特征;通过有效扩增测定10个太平洋海马样品的COI和ATP6基因片段序列,分别分析两种序列中碱基组成、变异位点等信息;基于K2P模型计算太平洋海马的种内种间遗传距离;通过NJ法构建太平洋海马与常见市售海马的系统聚类树,比较各海马间的亲缘关系。结果:太平洋海马最典型的性状特征包括体表光滑和体型巨大,全身密布白色竖线纹和突出的眼棘;10条太平洋海马样品COI、ATP6序列平均长度分别为649和603 bp,变异率分别为1.079%和1.990%。COI和ATP6序列的最大种内遗传距离均显著小于最小种间遗传距离,因此能与其他海马明显区分,同时与大海马的亲缘关系最近。结论:COI和ATP6序列均可作为将太平洋海马与其它常见海马明显区分的有效条形码,但ATP6序列更适合作为其标准条形码。本文提供了太平洋海马的性状和分子鉴别特征,为其品种的快速鉴定提供了技术支撑,对有效维护海马药材的市场秩序,控制和提高海马药材质量具有重要意义。
Objective:To establish the pharmacognosy identification methods of Hippocampus ingens using the combination of morphological characteristics and DNA barcodes,which could contribute to the development and application for further seahorse research.Methods:The typical characteristics of H.ingenswere obtained based on morphological observation according to classification and identification methods in traditional pharmacognosy research.After the amplification and sequencing of COI and ATP6 sequences from seahorse samples,the base composition,GC content,variable sitesand other information were calculated and compared,respectively.The intraspecific and interspecific variations of H.ingenswere determined based on Kimura 2-parameter model.The phylogenic tree of H.ingens and other common Hippocampus species was constructed by Neighbor-Joining method.Results:The most typical features of H.ingens includes smooth surface,large body,prominent eye spine and white vertical lines on whole body.The length of the measured COI and ATP6 sequences were 649 and 603 bp,respectively,with the mutation rate of 1.079%and 1.990%.The maximum intraspecific distances of both COI and ATP6 in H.ingens were much lower than the respective corresponding minimum interspecific distance,which indicated that H.ingens could be differedfrom other seahorses,and was most closely related to H.kuda.Conclusion:The DNA barcodes COI and ATP6 sequences could distinguish H.ingens from other seahorses accurately,and ATP6 sequences were more suitable to be used as standard DNA barcode for H.ingens identification.This paper illustrated the key features of morphological characteristics and molecular identification features of H.ingens,which provided technical support for rapid identification of the specie.It also has significant impacts for the improving the thequality of hippocampus species.
作者
来梦茹
孙思雅
方昀
陈建真
葛宇清
张光霁
程汝滨
Lai Mengru;Sun Siya;Fang Yun;Chen Jianzhen;Ge Yuqing;Zhang Guangji;Cheng Rubin(College of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou 310053,China;The First Affiliated Hospital,Zhejiang Chinese Medical University,Hangzhou 310053,China)
出处
《成都中医药大学学报》
2021年第4期37-42,共6页
Journal of Chengdu University of Traditional Chinese Medicine
基金
国家中医药管理局公益性行业科研专项经费项目(201507002-4-4)
浙江省新苗人才计划项目(752214F00803)。
关键词
太平洋海马
性状鉴别
DNA条形码
COI
ATP6
Hippocampus ingens
Morphological characteristics identification
DNA barcode
COI
ATP6
作者简介
第一作者:来梦茹,女,1996年生,硕士研究生在读,E-mail:617593969@qq.com;通信作者:程汝滨,男,1982年生,博士,副研究员,E-mail:biothcheng@hotmail.com。
