ABSTRACT
OBJECTIVE: To identify Belamcandae Rhizoma, Iridis Tectori Rhizoma and their adulterants by ISSR and SCoT markers. METHODS: The genome of Belamcandae Rhizoma, Iridis Tectori Rhizoma and its adulterants were amplified by the optimized ISSR and SCoT PCR conditions and screened primers. Genome polymorphism analysis and the dendrogram construction were calculated by the POPGENE 1.32 and the NTSYS-pc version 2.10 respectively. RESULTS: The 130 and 143 bands were amplified by the screened ISSR and SCoT primers respectively. The percentage of polymorphic bands were 96.2% and 97.9% respectively. Shannon diversity index(I) were 0.540 2 and 0.500 3, Nei's gene diversity index (H) were 0.363 0 and 0.327 3 respectively. The genetic distance calculated based on SCoT marker was higher than that of ISSR marker, which suggested that Elamcandae Rhizoma, Iridis Tectori Rhizoma and their adulterants have great genetic diversity. Cluster analysis based on SCoT genetic similarity indicated that Belamcandae Rhizoma, Iridis Tectori Rhizoma formed independent group with far relationship among their adulterants. CONCLUSION: Belamcandae Rhizoma, Iridis Tectori Rhizoma and their adulterants could be distinguished stablely, fastly and clearly by SCoT marker. The ISSR marker could only be used for identification of Belamcandae Rhizoma and its adulterants. The SCoT marker is better suitable for genetic diversity research on the low taxonomic category in genus Iris.
ABSTRACT
This study aimed to identify Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants by ITS2 sequence. All the DNA samples of Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants were extracted. The ITS2 sequence were succesfully amplified, and purified PCR products were sequenced. All the sequences were assembled using the CondonCode Aligner V3.7.1. The Kimura 2-Parameter (K2P) genetic distances and the Neighbor-Joining (NJ) phylogenetic tree were calculated by using MEGA5.1. Results indicated that the maximum intraspecific genetic distances of Belamcandae Rhizoma was 0, and the average GC content was 52.22%;the maximum intraspecific genetic distances of Iridis tectori Rhizoma was 0.004, and the average GC content was 67.87%. The maximum K2P intraspecific genetic distance of Belamcanda chinensis, Iris tectorum were both lower than the minimum interspecific genetic distance of adulterants. Additionally, the ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. The NJ tree based on ITS2 sequence indicated that Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants could be distinguished clearly. It could be concluded that ITS2 barcode can be used to correctly identify Belamcandae Rhizoma, Iridis tectori Rhizoma from their adulterants, and ensure their safety in use.