目的 用ITS 全序列鑒別各地產(chǎn)陽春砂及常見偽品。方法 從各地產(chǎn)陽春砂及常見偽品綠殼砂和海南砂中提取總DNA,以核基因組通用引物ITS 為引物進行擴增,擴增產(chǎn)物經(jīng)純化后,用PCR 產(chǎn)物直接法進行測序。結果 各樣品的ITS 序列總長度均為626bp,其中ITS-1序列長度為248bp,5.8S 序列長度為155bp,ITS-2序列長度為223bp;6個陽春砂和綠殼砂的ITS-2序列完全一致,各樣品的5.8S 序列完全一致,在ITS-1中各樣品卻有不同的堿基位點。結論 ITS-1序列可對陽春砂的道地性作出鑒別,明顯區(qū)分其偽品。

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.

國家中醫(yī)藥管理局科研基金(950379);廣東省自然科學基金(950615)