摘要
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： To evaluate the identification ability of some current DNA universal sequences in curcuma kwangsiensis with DNA barcoding technology to solve the problem which is difficult to identify curcuma kwangsiensis because of its wide varieties and similar characters. Methods： The PCR of curcuma kwangsiensis was amplified and sequenced by the universal primers of ITS, ITS2, rpoB, rpoC1, matK and psbA-trnB sequence. The successful rate of amplification and sequence and the variation between intraspecific, interspecific crossing and barcoding gap were compared. The identification ability of different sequences were evaluated by the BLAST 1 method. Results： The PCR amplification and sequence rate of curcuma kwangsiensis achieved 100% by the primer of ITS2. There were obvious advantages on the variation between intraspecific and interspecific crossing and barcoding gap compared to other universal sequences of DNA barcodes. The successful rate of identification achieved 100%. The successful rate of sequence and identification achieved over 90% by the primer of matK, rpoCl and rpoB, but the variations between intraspecific and interspecific crossing were not obvious. The barcoding gap of ITS was obvious, but the overlapping ratio of the variations between intraspecific and interspecific crossing was large and the successful rates of identification and sequence were low. The successful rates of amplification and sequence by the primer of psbA-trnH were lower than those by the primer of ITS2, but the successful rate of identification achieved 100% and the variations between intraspecific and interspecific crossing was obvious. Conclusion： ITS2 and pshA-trnH sequences can precisely identify the curcuma kwangsiensis. ITS2 may be a potential sequence of DNA barcode for identifying curcuma kwangsiensis, while psbA- trnH as a complement senquence.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2015年第1期100-103,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
广西科学研究与技术开发计划项目(No.桂科攻11107010-3-2)
桂林市科学研究与技术开发计划项目(No.20120105-7)
广西医药产业人才小高地项目(No.1305)~~
