目的 基于藁本屬Ligusticum葉綠體基因組高頻插入缺失區(qū)域開發(fā)InDel標記，同時結(jié)合通用條形碼對道地川芎及其常用混偽品進行種質(zhì)鑒定和系統(tǒng)發(fā)育研究。方法 通過8個DNA通用條形碼ycf1、matK、ITS2、rpoC1、rbcL、rpoB、trnK、psbA-trnH序列片段對采集的26個川芎樣品及其常用混偽品進行了擴增和測序，采用了遺傳距離統(tǒng)計法、barcoding gap分析法和構(gòu)建系統(tǒng)發(fā)育樹法進行親緣關(guān)系和系統(tǒng)發(fā)育研究。同時利用InDel分子標記，采用構(gòu)建進化樹法對道地川芎及其混偽品物種進行分子鑒定。結(jié)果 rbcL片段保守位點最多（97.32%），且GC含量最高（44.9%）。rbcL+rpoB片段具有最小的平均種內(nèi)遺傳距離（0.002 5），psbA-trnH片段具有最大的平均種間遺傳距離（0.429 2）。trnK片段和rbcL+rpoB片段具有最高的種間變異，psbA-trnH片段的“barcodinggap”區(qū)重疊度最小。采用的8對DNA通用條形碼無法準確地鑒定道地川芎藥材與其他混偽品物種。InDel分子標記的聚類分析中，24對InDel引物中的4對引物組合能準確地鑒定出道地川芎，并將道地川芎及其混偽品物種聚類為4個分支，其中一個分支為采集的道地川產(chǎn)川芎藥材。結(jié)論 InDel標記對道地川芎及其常用混偽品的鑒定能力高于通用條形碼。對于傳統(tǒng)的通用DNA條形碼，由于遺傳成分差異大，無法區(qū)分川芎及其常用混偽品。新開發(fā)出的InDel分子標記可以有效地鑒定道地川芎及其常用混偽品，從分子水平上為川芎道地性研究提供有效手段。

Objective In this study, InDel markers were developed based on the high frequency Insertion/Deletion region of chloroplast genome of Ligusticum. Germplasm identification and phylogenetic development of Ligusticum chuanxiong and its common adulterants were studied with universal barcode. Methods The 26 samples of L. chuanxiong and its common adulterants were amplified and sequenced by eight DNA universal barcodes:ycf1, matK, ITS2, rpoC1, rbcL, rpoB, trnK, and psbA-trnH. Genetic distance statistics, barcoding gap and phylogenetic tree analysis methods were used to study the phylogenetic relationship and phylogeny of L. chuanxiong. At the same time, the evolutionary tree was constructed to study molecular identification of L. chuanxiong and its common adulterants. Results The results showed that rbcL conserved site was the highest (97.32%) with the highest GC content (44.9%). The rbcL+rpoB fragment had the smallest average intraspecific genetic distance (0.002 5). The psbA-trnH sequence fragment had the largest average interspecific genetic distance (0.429 2). The trnK and rbcL+rpoB sequence had the highest interspecific genetic distance. The overlap of the "barcoding gap" region of psbA-trnH was the least. The species of L. chuanxiong and other adulterated species were not accurately identified by the eight pairs of DNA barcodes. The cluster analysis of 24 InDel markers could accurately identify genuine L. chuanxiong and classify the species of L. chuanxiong and its adulterants into four categories, one of which was genuine L. chuanxiong collected from Sichuan. Conclusion The ability of InDel markers to identify authentic L. chuanxiong and its common adulterants was higher than that of common barcode. According to the above studies, it is found that it is impossible to distinguish L. chuanxiong and its common adulterants by the traditional DNA barcodes because of the large difference in genetic components. The newly developed InDel molecular markers can effectively identify L. chuanxiong and its commonly used adulterants, and provide an effective method for the genuineness of L. chuanxiong at molecular level.

四川省育種攻關(guān)（2016WY0036-4-1）；四川省財政創(chuàng)新提升工程（2016TSCY-001）；四川農(nóng)業(yè)科學院優(yōu)秀論文基金（2018LWJJ-019）