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《China Journal of Chinese Materia Medica》 2007-08
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Molecular identity of Crocus sativus and its misused substitutes by ITS sequence

CHE Jian,TANG Lin,LIU Yan-jun,HE Wei,CHEN Fang(College of Life Science,Sichuan University,Chengdu 610064,China)  
Objective:To find the patterns of the rDNA ITS sequence variation of Crocus sativus,Chrysanthemum chanetii,Nelumbo nucifera,Zea mays and Garthamus tinctorius and to establish the molecular biological method for the identification of C.sativus and the others. Method:After the total DNA of Crocus sativus,C.vernus-w and C.vernus-p were extracted,the ITS sequence was amplified by PCR with universal primer of ITS and PCR product was sequenced after purification and cloning.The ITS sequences of Chrysanthemum chanetii,Nelumbo nucifera,Zea mays and Garthamus tinctorius were obtained from GenBank.Result:The complete ITS sequence of Crocus sativus,C.vernus-w and C.vernus-p,including ITS1 rDNA,5.8S rDNA,ITS2 rDNA were measured.The GenBank accession No.was DQ094185,DQ224363 and DQ224364 respectively. The similarity of ITS sequence between C.sativus and the two garden species of C.vernus was above 91%;the identity was 99.84% between C.vernus-w and C.vernus-p.The range of diversity between C.sativus and other herbs was above 46% based on ITS1 and above41% based on ITS2.Conclusion:C.sativus can be distinguished from misused substitutes by the ITS sequence.The ITS sequence is an available molecular marker for identification of the C.sativus.
【Fund】: 四川省应用基础研究计划资助项目(03-TY029090-2)
【CateGory Index】: S567.219
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