摘要
目的筛选与评价适用于云南草果居群的DNA条形码。方法以云南省草果种质资源为样本对ITS、psbA-trnH、matK、rbcL和ycf15条DNA条形码常用序列进行筛选与评价,并对草果居群进行扩增,测序,测序序列用Genestar进行拼接,然后用Mega进行数据处理,并对草果多样性及其鉴定进行分析。结果引物ITS5和ITS4对草果的扩增片段长度大约为520 bp;rbcLa-F和rbcLa-R对草果的扩增片段长度大约为498 bp;引物ycf1-bF和ycf1-bR对草果的扩增片段长度大约为800 bp;引物psb A-trn H-1F和psbA-trnH-1R对草果的扩增片段长度大约为400 bp;引物matK-2F和matK-2R对草果的扩增片段长度大约为470 bp。扩增及测序的成功率均较高,结果大多可用。通过对草果ITS、psb A-trnH、matK和ycf1序列的扩增结果进行分析,草果与其他豆蔻属植物都可以被清晰地区分开;ITS序列所有样本分为MG5白花草果居群和其他居群;psb A-trn H序列所有样本分为MG5白花草果居群,MG6黄花草果居群和其他居群;matK序列所有样本分为MG6黄花草果居群和其他居群,MG5白花草果样本扩增失败;ycf1序列所有样本分为MG6黄花草果居群和其他居群,MG5白花草果居群与其他22个草果居群聚为一支;rbcL序列对所有样本的扩增均一致。结论ITS、matK、psb A-trn H及ycf1序列均能将草果与其他同属植物进行准确区分;MG6的matK、psbA-trnH及ycf1序列发现了序列位点的变异,为草果品种的选育做出贡献。ITS和psb A-trn H序列可将黄花和白花草果序列区分开;草果白花黄花所有样本rbcL序列无任何变异,且用rbcL序列无法鉴别草果与其他同属植物,可将其舍去。
Objective To screen and evaluate DNA barcoding of Amomun tsao-ko populations in Yunnan.Methods ITS,psbA-tmH,matK,rbcL,and ycfl sequences were screened and evaluated using A.tsao-ko as samples.The samples of A.tsao-ko population were amplified and sequenced.The sequences were spliced with Genestar,and then processed with Mega for data processing.And A.tsao-ko diversity and identification were analyzed and discussed.Results The length of the amplified fragments of primers ITS5 and ITS4 was approximately 520 bp;The length of the amplified fragments of the primers rbcLa-F and rbcLa-R was approximately 498 bp;The length of the amplified fragments of the primers ycfl-bF and ycfl-bR was approximately 800 bp;The length of the amplified fragments of the primers psbA-tmH-IF and psbA-tmH-1R was approximately 400 bp;The length of the amplified fragments of the primers matK-2F and matK-2R was approximately 470 bp.The success rate of amplification and sequencing was high,and most of the results were available.By analyzing the amplification results of ITS,psbA-tmH,matK and ycfl sequences of A.tsao-ko,A.tsao-ko and other Amomum genus plants can be clearly distinguished;All samples of the ITS sequence were divided into MG5 white flower A.tsao-ko population and other populations;All samples of the psbA-tmH sequence were divided into MG5 white flower A.tsao-ko population,MG6 yellow flower A.tsao-ko population and other populations;All samples of the matK sequence were divided into MG6 A.tsao-ko population and other populations.The MG5 white flower A.tsao-ko sample failed to be amplified;All samples of the ycfl sequence were divided into the MG6 yellow flower A.tsao-ko population and other populations,and the MG5 white flower A.tsao-ko population was clustered with the other 22 A.tsao-ko populations;The amplification of rbcL sequence was consistent for all samples.Conclusion The ITS,matK,psbA-tmH and ycfl sequences can accurately distinguish A.tsao-ko from other plants of Amomum genus;The sequence site variations were found in matK,psbA-tmH and ycfl sequences of MG6.This research has contributed to the selection and breeding of A.tsao-ko varieties.ITS and psbA-tmHsequences can distinguish yellow flower and white flower of A.tsao-ko;There is no variation in the rbcL sequence of all samples of white and yellow flowers of A.tsao-ko,and Amomum tsao-ko and other plants of Amomum genus cannot be identified with the rbcL sequence,which can be discarded.
作者
胡一凡
张雪梅
石乃星
杨志清
HU Y i-fan;ZHANG Xue-mei;SHI Nai-xing;YANG Zhi-qing(College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;Yunnan Key Laboratory of Medicinal Plant Biology,Yunnan Agricultural University,Kunming 650201,China;National&Local Joint Engineering Research Center on Germplasm Innovation&Utilization of Chinese Medicinal Materials in Southwestern China,Yunnan Agricultural University,Kunming 650201,China;Yunnan Research Center for Aromatic Biology Engineering Technology,Kunming 650201,China)
出处
《中草药》
CAS
CSCD
北大核心
2019年第24期6091-6097,共7页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金项目：草果居群的分子鉴定及种质资源评价(81560615)
云南省社会发展科技计划项目：云南草果种植加工关键技术研究与示范(2011CG015)
云南省科技计划项目：云南省芳香生物工程技术研究中心建设(2018DH010)
