Allele-specific diagnostic PCR authentication of Dendrobium huoshanense and its allied species of Dendrobium Sw / 中草药

ABSTRACT

Objective To design a pair of allele-specific diagnostic primer for authenticating Dendrobium huoshanense from other species of Dendrobium Sw.only by PCR.Methods Based on rDNA ITS sequences database of D.huoshanense and other species of Dendrobium Sw.,an allele-specific diagnostic primer pairs was designed.Diagnostic PCRs were performed using the primer with the total DNAs of 19(original) plants from Dendrobium Sw.as templates.The positive was the genuine.Results When the(annealing) temperature was raised to 60 ℃, the template DNA of D.huoshanense could be amplified whereas the diagnostic PCR of the rest species were all negative.Conclusion The diagnostic PCRs can authenticate D.huoshanense from other species of Dendrobium Sw.efficiently.Compared to the authentication method by sequencing DNA fragments and morphological traits etc.,the allele-specific diagnostic PCR is not only simple with time-saving but also practical and effective.