The Applications Of DNA Sequences And Inter-simple Sequence Repeats For Authenticating Herbs

A few of wild populations of Chinese traditional medicine were authenticated with the sequences of rDNA ITS,and their relationship was studied in this study.The sequences of rDNA ITS region of Zanthexylum bungeanum ranged from 619 to 620 bp, and the length difference amoung Zanthexylum bungeanum and their adulterants is 4 bp. There are 15 variable sites, 12 informative sites and 3 authenticable sites among Zanthexylum bungeanum populations. The difference of rDNA ITS regions amoung Zanthexylum bungeanum and their adulterants are obvious, the number of variable sites is 71. The differences of rDNA ITS sequences can be used to authenticate accurately the populations of Zanthexylum bungeanum and their adulterants. Those populations of Z. bungeanum which have close relationship always distribute in near geographic areas. The sequences of rDNA ITS region of Alisma orientale ranged 640 bp.The sequence of Alisma orientale from Jiangxi and Fujian provinces was the same ,and there were 2 stable variable sites among Alisma orientale populations which were collected from Sichuan and Fujian(Jiangxi) provinces. One was in ITS-1, and the other was in ITS-2. The difference of rDNA ITS sequences can be used to authenticate accurately the populations of Alisma orientale from Sichuan and Fujian, and to provide reliable molecular markers for identifying Alisma orientale populations.Zanthexylum bungeanum and its fake oens were authenticated with the sequences of cpDNA trnL-F. Length of cpDNA trnL-F of Zanthexylum bungeanum Maxim is 370 bp with 62.2 percent of AT sites.The difference is obvious between Zanthexylum bungeanum Maxim and fake ones in their cpDNA trnL-F regions though there's little diversity from their appearance of fruits together with physical characters. While this internal space has high alikeness among taxa classified by genus below, reaching full percent parallel. The cpDNA trnL-F sequences are only suitable for the authentication of Zanthexylum bungeanum Maxim from fake ones on the basis of Genus above.ISSR-PCR system of Dendrobium officinale Kimura et Migo was established and optimized according to the ISSR-PCR characters of Dendrobium officinale. ISSR-PCR reaction was put up using 10 out of 76 ISSR primers which could generate reproducible polymorphic fragments. A total of 127 DNA fragments were amplified by 10 primers,of which 115 was polymorphic and the percentage of polymorphic bands is 90.3%. 9 polymorphic fragments which amplified by 5primers could be used as authentic markers and 8 populations of Dendrobium officinale Kimura et Migo could be identified and relevant identification code of ISSR molecular marker was established.