Xuexia Li

Genetic variation and conservation of the endangered Chinese endemic herb Dendrobium officinale based on SRAP analysis

Dendrobium officinale (Orchidaceae) is used for traditional medicine and is critically endangered in China. To investigate the genetic structure of this species and to offer some advice on conservation strategies, 84 individuals from nine wild populations of D. officinale were analyzed using the method of sequence-related amplified polymorphism. A high level of genetic diversity was detected (PPB = 88.07%, HE = 0.2880) at the species level. However, the genetic diversity at the population level was lower (PPB = 51.68%, HE = 0.1878) in comparison with other species with similar life history characteristics. Based on analysis of molecular variation, there was moderate variation between pairs of populations with ΦST values ranging from 0.1327 to 0.4151 and on average 27.05% of the genetic variation occurred among populations. Two main clusters were shown in UPGMA using TFPGA, which is consistent with the result of principal coordinate analysis using NTSYS. In situ conservation is the first advocated and and ex situ should be proposed at the same time to protect the endangered plant and to preserve germplasm resources.

Genetic diversity across natural populations of Dendrobium officinale, the endangered medicinal herb endemic to China, revealed by ISSR and RAPD markers

Dendrobium officinale is a rare and endangered herb with special habitats and endemic to China. Genetic diversity was examined within and among nine natural populations using inter-simple sequence repeat (ISSR) and random amplified polymorphic (RAPD) for conservation. Both molecular markers revealed a high percentage (>89%) of polymorphic bands and ISSR markers detected more diversity than RAPD markers. Analysis of molecular variance (AMOVA) revealed that 78.84% (ISSR) and 78.88% (RAPD) of variability was partitioned among individuals within populations. This genetic structure was probably due to severe genetic drift resulting from habitat fragmentation and human overexploitation since 1950s. Moreover, there is a lack of significant association between genetic and geographic distances (r = 0.276; p > 0.05) in the populations of D. officinale. From the conservation point of view, populations GL, GS and GSD with higher genetic diversity should be protected firstly to maintain the species potential for evolutionary change and population YG with lower diversity but representing a novel evolutionary unit should also be paid more attention to during D. officinale conservation practice.

Molecular Authentication of Dendrobium loddigesii Rolfe by Amplification Refractory Mutation System (ARMS)

As a widely used medicinal plant, Dendrobium loddigesii Rolfe is always a possible target for fraudulent labeling. The identification of D. loddigesii is generally difficult from its morphological and chemical appearance only. In order to develop a convenient and efficient identification method for D. loddigesii, six pairs of diagnostic ARMS primers were designed based on nrDNA ITS sequences of D. loddigesii and eleven adulterants. The results showed that one diagnostic primer pair (FJB-04-forward, FJB-04-reverse) could be used to authenticate D. loddigesii by generating a fragment of 353 bp at annealing temperatures from 48 degrees C to 62 degrees C while the other diagnostic primer pair (FJB-03-forward, FJB-03-reverse) took on the same effect at annealing temperatures from 49 degrees C to 55 degrees C. This points out the potential of ARMS analysis for authentication of D. loddigesii.

Development of microsatellite markers in Dendrobium fimbriatum Hook, an endangered Chinese endemic herb

As an endangered endemic herb, Dendrobium fimbriatum, is under threat from numerous impacts. In order to analyse the genetic diversity and structure of this endangered species, we provide details of 10 microsatellite loci (out of 15 primer pairs designed) which showed polymorphic for D. fimbriatum. These loci were used to screen 25 individuals from across the species’ geographical range. Ten loci were polymorphic with 2 to 19 alleles; three loci were monomorphic, while the rest produced no amplification fragments. These loci will be used to investigate population genetic structure, genetic diversity, conservation, and individual authentication in the endangered D. fimbriatum.