基于matK+rbcL+trnH-psbA联合分析法对罂粟属的植物鉴定

  目的  利用DNA条形码技术探索罂粟的鉴定方法，寻找可靠的DNA条形码序列对罂粟属植物进行种内和种间的鉴别。   方法   采用5种不同种类的虞美人和花菱草作为罂粟鉴别的种内植物、种间植物。利用叶绿体基因组matK、rbcL、trnH-psbA和核糖体基因组ITS2、ITS4、ITS5引物对提取的DNA进行扩增，并双向测序拼接，利用MEGA.X.对序列进行分析比对，计算种内、种间遗传距离，同时使用该软件构建NJ系统发育树。   结果   将测序结果录入Genbank中进行比对分析，结果显示单一DNA条形码序列无法有效区分虞美人与罂粟。通过Genbank下载数据使用matK+rbcL+trnH-psbA组合条形码构建系统发育树成功，对罂粟和虞美人样本测序结果进行验证。经验证，matK+rbcL+trnH-psbA组合条形码构建NJ发育树成功，能有效区分虞美人和罂粟。   结论   matK+rbcL+trnH-psbA作为组合DNA条形码使用，可以有效鉴别罂粟属内植物。

Identification of Papaveraceae Based on Chloroplast DNA matK+rbcL+ trnH-psbA

  Objective   To explore the identification method of Papaver with the use of DNA barcoding technology and find reliable DNA barcoding sequence for intraspecific and interspecific identification of Papaver.   Methods   Five different species of Papaver rhoeas and Eschscholtzia californica Cham were used as intraspecific and interspecific plants for the identification of Papaver. The extracted DNA was amplified using primers matK, rbcL, trnH-psbA from chloroplast genomes and ITS2, ITS4, ITS5 from ribosomal genomes, and then bidirectional sequencing and splicing were performed using MEGA X. Sequences were analyzed and aligned, intraspecific and interspecific genetic distances were calculated, and N-J phylogenetic trees were constructed using the software.   Results  The sequencing results, which were entered into Genbank for comparative analysis, showed that a single DNA barcode sequence was not effective in distinguishing between Papaver somniferum and Papaver rhoeas. At the same time, a phylogenetic tree was successfully constructed using matK+rbcL+trnH-psbA combined barcode through Genbank download data, and the sequencing results of Papaver samples in this study were verified by this method. The combination of matK+rbcL+trnH-psbA barcode successfully constructed the N-J tree, which could effectively distinguish between Papaver somniferum and Papaver rhoeas in this study.   Conclusion  matK+rbcL+ trnH-psbA can be used as a combined DNA barcode for the identification of Papaver.