Isolation and characterization of microsatellite markers in Dendrobium officinale, an endangered herb endemic to China

Abstract

As an invaluable herb, Dendrobium officinale has been in severe danger since 1950 because of human exploitation and habitat deterioration. In order to investigate the genetic diversity and structure of this species, we isolated and characterized 12 microsatellite loci derived from two microsatellite-enriched libraries. Twenty-two individuals from Leye population were analysed. These loci were polymorphic and displayed three to 12 alleles per locus. The observed and expected heterozygosities ranged from 0.150 to 0.624 and from 0.162 to 0.605, respectively. These are the first microsatellite loci characterized from D. officinale that will contribute to research on the conservation, genetic diversity, population structure and individual authentication.

Identification of the Geographic Origin of <i>Dendrobium thyrsiflorum</i> on Chinese Herbal Medicine Market Using Trinucleotide Microsatellite Markers

“…The mean expected heterozygosity across loci within populations of D. thyrsiflorum ranged from 0.76 to 0.82, suggesting that this species has a high level of genetic diversity comparable to that of other Dendrobium species. 10,33,34) Based on a published study, 12) a high level of genetic diversity of D. thyrsiflorum may increase the power of the statistical analysis for identifying the origin of living or processed D. thyrsiflorum samples.…”

“…Numerous lines of evidence have demonstrated that SSRs exist in any region of eukaryotic genomes, including coding and non-coding regions. Microsatellite markers have been used to identify the geographic origins of commercially available herbs 10) and shown to be an effective tool for individual assignment [11][12][13][14][15] as well as genetic variation analysis for various species 16,17) due to their features: composite genotypes across several loci, relatively high polymorphism, and co-dominant inheritance. 15,18) On the other side, microsatellite markers are quite species-specific, which limits their use as a universal tool for testing multiple species.…”

“…Subsequently, super SNX24 linkers (AP11, 5Ј-GATCGTCGACGGTACCGAATTCT-3Ј and AP12, 5Ј-GT-CAAGAATTCGGTACCGTCGAC-3Ј) serving as priming sites for polymerase chain reaction (PCR) were ligated onto the ends of the fragments. The enrichment of DNA fragments was performed by their hybridization with biotinylated oligonucleotide probes (GAA) 10 , and then streptavidincoated magnetic beads (Promega, U.S.A.) were used to capture biotinylated oligos. After stringent washing, the captured DNA fragments were amplified using SNX24 primers and inserted into pMD18-T using the TA cloning kit (TaKaRa) and subsequently transformed into chemically competent cells (DH5a).…”

Isolation and characterization of novel EST-SSRs in the showy dendrobium, Dendrobium nobile (Orchidaceae)

“…The primers were used to amplify SSR loci in D. nobile genomic DNA, and the amplified bands with bands that were clear and the expected size were sequenced to validate the corresponding repeat motifs. All of the sequences of the developed D. nobile SSRs were compared with the Dendrobium SSRs that were previously reported (Gu et al, 2007;Xie et al, 2010;Lu et al, 2012Lu et al, , 2013 in order to determine that they were novel.…”

“…However, as a non-model plant that lacks a sequenced genome, thus far only over a hundred Dendrobium simple sequence repeat (SSR) markers have been reported (Gu et al, 2007;Xie et al, 2010;Lu et al, 2012a,b). Here, we present the development of novel polymorphic expressed sequence tag (EST)-SSRs in D. nobile, which will be helpful for potential genetic applications.…”