云南草果种质资源DNA条形码序列分析

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ycf1 5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psbA-trnH-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、psbA-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；psbA-trnH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、psbA-trnH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psbA-trnH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。

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

Objective To screen and evaluate DNA barcoding of Amomun tsao-ko populations in Yunnan. Methods ITS, psbA-trnH, matK, rbcL, and ycf1 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 ycf1-bF and ycf1-bR was approximately 800 bp; The length of the amplified fragments of the primers psbA-trnH-1F and psbA-trnH-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-trnH, matK and ycf1 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-trnH 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 ycf1 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-trnH and ycf1 sequences can accurately distinguish A. tsao-ko from other plants of Amomum genus; The sequence site variations were found in matK, psbA-trnH and ycf1 sequences of MG6. This research has contributed to the selection and breeding of A. tsao-ko varieties. ITS and psbA-trnHsequences 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.