Thirteen novel polymorphic microsatellite markers for endangered Chinese endemic herb Dendrobium Officinale
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TECHNICAL NOTE
Thirteen novel polymorphic microsatellite markers for endangered Chinese endemic herb Dendrobium Officinale Jiang-Jie Lu • Ling Gao • Jie-Yu Kang • Shang-Guo Feng • Ren-Feng He • Hui-Zhong Wang
Received: 10 October 2012 / Accepted: 29 October 2012 / Published online: 7 November 2012 Ó Springer Science+Business Media Dordrecht 2012
Abstract We isolated and developed 13 novel microsatellite makers from expressed sequence tag sequences of endangered Chinese endemic herb Dendrobium officinale and examined them for 38 individuals from Jinhua, Zhejiang, Chian. These loci displayed the mean number of 4. 31 alleles per locus, and with expected heterozygosity (He) and observed heterozygosity (Ho) per locus ranged from 0.4438 to 0.6842 and from 0 to 0.4211, respectively. These markers have potential for application in germplasm appraisal, genetic diversity study, genetic mapping, and molecular breeding in D. officinale and other congeneric species. Keywords Dendrobium officinale Microsatellite Expressed sequence tag (EST)
Dendrobium officinale Kimura & Migo, along with D. chrysanthum Wall. ex Lindl., D. fimbriatum Hook., D. loddigesii Rolfe, D. nobile Lindl., were listed in the Chinese Pharmacopoeia, has been acknowledged for its unparalleled curative effects (Chinese Pharmacopoeia Editorial Committee 2000). D. officinale is subjected to mass-collection for a long time, the habitat deterioration and over exploitation enticed by high profits has led to the wild germplasm resources depleted seriously in recent years, and now they are under first-grade state protection in China. For effective conservation and utilization of
J.-J. Lu L. Gao J.-Y. Kang S.-G. Feng R.-F. He H.-Z. Wang (&) Zhejiang Provincial Key Laboratory for Genetic Improvement and Quality Control of Medicinal Plants, Hangzhou Normal University, Hangzhou 310018, China e-mail: [email protected]
D. officinale resource, it is necessary to investigate the genetic diversity and the population genetic structure by using various types of DNA markers, especially desirable microsatellite markers (Kalia et al. 2010). Nevertheless, the development of microsatellite markers in non-model plant D. officinale has been very slow placed limits on the lack of genome sequences. Up to date, little microsatellite markers of this endangered Chinese endemic herb have been reported (Gu et al. 2007; Xu et al. 2011). A total of 800 ESTs sequences were derived from GenBank (http://www.ncbi.nlm.nih.gov/) for D. officianle, followed by detecting the tandem repeats of 2-6 nucleotide with the web software Microsatellite Repeats Finder (http://biophp.org/minitools/microsatellite_repeats_finder), 92 candidate microsatellite loci were found. By using Primer 3.0 software (Rozen and Skaletsky 2000), 30 pairs of primers (length of 20 ± 2 nucleotide, GC content of 40-60 %, optimum annealing temperature of at least 50 °C, and PCR product size ranging from 100 to 300 bp) were designed. The primers were used to amplify the microsatellite loci in D. officinale genome DNA, and
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