Identification Of Alpinia Officinarum Hance And Alpinia Galanga (L.)Swartz By Sequence Analysis Of The RDNA Internal Transcribed Spacer Regions

Objective:To analyze the genetic differentiation of rDNA ITS sequences among the geographical population of Alpinia officinarum Hance and its counterfeit species after sequencing the PCR products of rDNA ITS regions, and to provide melocular markers for identifying Alpinia officinarum Hance against its closely related species. Methods:(1)The genomic DNA of Alpinia officinarum Hance populations and their adulterants was extracted by modified CTAB method.(2)Products were directly sequenced after amplifying the rDNA ITS sequence using consensus primer pairs. (3)Analyzed by means of Clustal X and MEGA2.0 sequence analysis software, Phylogenetic tree based on ITS sequences data was conducted by Neighbor joining method, Then the rDNA ITS sequences of Alpinia officinarum Hance populations and their adulterants were submitted to Genbank.Results:(1)It was indicated that the resultant DNA obtained by improved CTAB method was very high purity, after amplifited and searched with Blast methods.(2)Sequence analysis showed that Alpinia officinarum Hance length of ITS1 and ITS2 were 117 bp,227 bp; Alpinia galanga (L.)Swartz length of ITS1 and ITS2 were 178 bp,but 5.8s area were all 164 bp. the eight kinds of plants with a total length ITS sequence about 568-576bp, with 32 variable sites,mainly concentrated in the ITS1 and ITS2 area, mostly in between the transversion AG. there were more single nucleotide variants in ITS1, while the ITS2 area had two large fragments of the missing, and more consistent performance differences between species,5.8s area had only one polymorphic. (3)By comparison of the rDNA ITS sequence analysis, rDNA ITS sequences were landed in the GenBank,. Log numbered GU097441 (Xuwen,Guangdong), GU097439 (Nanning, Guangxi Alpinia), GU09744 2 (Gaozhou), GU097440 (Nanjing, Fujian), GU097443 (Wanning, Hainan).The results of phylogenetic tree indicated the tested materials clould be grouped into two classes.the results show that eight kinds of plants with a total length ITS sequence for 568-576bp,have 32 variable sites,mainly concentrated in the ITS1 and ITS2 area. The results of phylogenetic tree indicated the tested materials clould be grouped into two classes. Conclusion:(1)The rDNA ITS sequences of Alpinia officinarum Hance and its counterfeit species were obtained for the first time and submitted to GenBank; (2)The rDNA ITS sequences analyzed that Alpinia officinarum Hance populations and their adulterants has a stable and large genetic diffierences. (3)The rDNA ITS sequence is a better molecular marker for idertification of Alpinia officinarum Hance populations and their adulterants.