云南草果种质资源DNA条形码序列分析  

DNA barcoding sequence analysis of Amomum tsao-ko germplasm resources in Yunnan Province

作　　者：胡一凡 张雪梅[1] 石乃星 杨志清[1,2,3,4] 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)

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]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 [3]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 [4]云南省芳香生物工程技术研究中心,云南昆明650201

出　　处：《中草药》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)

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,

关 键 词：草果 遗传多样性 亲缘关系 DNA条形码 ITS序列 psbA-trnH序列 MATK序列 ycf1序列 

分 类 号：R282.12[医药卫生—中药学]