以matK序列作为DNA条形码,对从国外截获的7种苍耳属植物进行物种区分鉴定研究,采用DNeasyPlant Mini Kit试剂盒进行总DNA的提取,应用通用引物对其matK基因进行扩增,测序得到7种苍耳属植物的matK序列,利用MEGA 5.1软件对这7种苍耳属植物的matK序列进行比对和分析并构建系统树,结果显示:matK序列能从基因位点层面对苍耳属植物进行区分鉴定。
[Objective] This study aimed to provide reference and reduce the workload for screening standard DNA barcoding genes of plants. [Method] Three DNA barcoding genes ITS, ITS2 and rbcL were amplified from seven Xanthium species under the same PCR condition： PCR amplification was started with initial denaturation at 95 ℃ for 4 min, followed by 35 cycles of denaturation at 94 ℃ for 30 s, annealing at 52 ℃ for 45 s, and extension at 72 ℃ for 45 s; the amplification was completed by holding the reaction mixture at 72 ℃ for 10 min to allow complete extension of PCR, and the PCR products were stored at 4 ℃. [Result] Three DNA barcoding genes ITS, ITS2 and rbcL were all amplified successfully. [Conclusion] This study indicates that PCR amplification conditions for DNA barcoding genes ITS,ITS2 and rbcL in plants may be consistent.
