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目的:莪术类品种繁多,形态相似,不易鉴别,利用DNA barcoding技术评价目前几个热点的DNA通用序列对广西莪术鉴别的能力。方法:本研究使用ITS、ITS2、rpo B、rpo C1、mat K、psb A-trn H序列的通用引物对广西莪术进行PCR扩增和测序,通过比较各序列的扩增和测序成功率、种内和种间的变异、barcoding gap,并采取相似性探索BLAST 1方法评价不同序列的鉴别能力。结果:ITS2序列在所研究的广西莪术中的扩增效率为100%,其种内种间变异、barcoding gap与其他DNA条形码候选序列相比具有明显的优势,ITS2序列在广西莪术中的鉴定成功率达到100%,mat K、rpo C1和rpo B虽然测序和鉴定成功率都在90%以上,但是其种内及种间差异不明显,ITS虽然barcoding gap明显,但种内和种间遗传变异重叠比例较大,且测序成功率及鉴定效率不理想,psb A-trn H虽然在扩增及测序方面不如ITS2,但是其鉴定成功率为100%,有明显的种内及种间差异。结论:ITS2与psb A-trn H序列能够准确鉴定广西莪术植物,ITS2可以作为广西莪术药用植物的候选DNA条形码序列,而psb A-trn H可作为ITS2的补充序列。
Objective: Curcuma variety, morphological similarities, not easy to identify, using DNA barcoding technology to assess the current hot spots of several common DNA sequences of Guangxi Curcuma identification ability. Methods: The universal primers of ITS, ITS2, rpo B, rpo C1, mat K and psb A-trn H were used to amplify and sequence Guangxi Curcuma. By comparing the success rate of each sequence amplification and sequencing, Internal and interspecific variation, barcoding gap, and exploratory similarity search BLAST 1 method to evaluate the ability of different sequences to discriminate. Results: The amplification efficiency of ITS2 sequence in the studied Curcuma zingiberensis was 100%. The intraspecific variation, barcoding gap and other DNA barcode candidate sequences had obvious advantages. ITS2 sequence was identified successfully in Curcuma Although the success rate of matK, rpo C1 and rpo B was above 90%, the intraspecific and interspecific differences were not obvious. Although ITS barcoding gap was obvious, intraspecific and interspecific genetic variation The overlap ratio was large, and the success rate of sequencing and identification efficiency were not satisfactory. Although psb A-trn H was not as good as ITS2 in amplification and sequencing, its identification success rate was 100%, with significant intraspecific and interspecific differences. CONCLUSIONS: ITS2 and psb A-trn H sequences can accurately identify the genus Curcuma, ITS2 can be used as a candidate DNA barcode sequence of the genus Curcuma, and psb A-trn H can be used as a complementary sequence to ITS2.