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Specific PCR Identification between Dendrobium huoshanense and Its Adulterants

??OBJECTIVE To design specific identification primers based on ribosomal DNA ITS sequences and accurate identification of Dendrobium huoshanensis and its adultments.METHODS Phylogenetic tree of D. huoshanense and D. officinale and D. henanense by maximum parsimony (MP) and Bayesian inference (BI) were constructed. Specific primers SH-CP9s/SH-CP9a, SH-CP25s/SH-CP25a, SH-CP29s/SH-CP29a for PCR identification of D. huoshanensis based on mutation sites on ITS sequences were designed.RESULTS In the phylogenetic tree, D. huoshanense and D. officinale and D. henanense were clustered into monophyletic group. The specific PCR reaction procedure of D. huoshanense was obtained by investigating the annealing temperature and the amount of DNA template, and different types of PCR and Taq enzymes. In the PCR product, D. huoshanense appearance of the band, and adultments with no band.CONCLUSION The ITS sequences can be used as DNA Barcoding identification of D. huoshanensis, and identify D. huoshanense and its adultments by the designed primers.