目的 用ITS 全序列鉴别各地产阳春砂及常见伪品。方法 从各地产阳春砂及常见伪品绿壳砂和海南砂中提取总DNA,以核基因组通用引物ITS 为引物进行扩增,扩增产物经纯化后,用PCR 产物直接法进行测序。结果 各样品的ITS 序列总长度均为626bp,其中ITS-1序列长度为248bp,5.8S 序列长度为155bp,ITS-2序列长度为223bp;6个阳春砂和绿壳砂的ITS-2序列完全一致,各样品的5.8S 序列完全一致,在ITS-1中各样品却有不同的碱基位点。结论 ITS-1序列可对阳春砂的道地性作出鉴别,明显区分其伪品。

Object To identity Amomum villosum Lour. from different producing area by ITS sequence analysis.Methods Firstly, total DNA of A. villosum from different producing area was extracted and it's succedaneum or fake was extracted. Secondly, the ITS sequence was amplified by PCR with universal primer of ITS and sequencing ITS on PCR product was directly sequenced after purification.Results The total length of ITS sequence is 626 bp in the different samples. The ITS sequence can be divided into three fragments: the length of ITS-1 is 248 bp, 5.8 S is 155 bp and ITS-2 is 223 bp respectively. In all the samples, 5.8 S has the same sequence. Six A. villosum from different producing area and Luqiao amomum have the same ITS-2 sequence. But ITS-1 fragment has bigger diversity in base sequence among the various specimens.Conclusion ITS-1 sequence can be used to confirm producing area of A. villosum, and find out its fake.

国家中医药管理局科研基金(950379);广东省自然科学基金(950615)