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Bulletin of Botanical Research ›› 2017, Vol. 37 ›› Issue (1): 78-87.doi: 10.7525/j.issn.1673-5102.2017.01.011

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Evaluation,Selection and Application of Dendrobium officinale EST-SSR Primers

HU Zhong-Yi, FU Tao, HE Yue-Qiu, LI Wen, LIN Li   

  1. Ningbo City College of Vocational Technology, Ningbo 315502
  • Received:2016-07-04 Online:2017-01-15 Published:2017-03-06
  • Supported by:
    Scientific research project of Education Department of Zhejiang Province(Y201533793);School Scientific Research Youth Special Topics of Ningbo City College of Vocational Technology(ZZX15064)

Abstract: By developing and evaluating EST-SSR primers, we studied their application in genetic diversity, species identification, genetic map construction, genetic relationship identification of Dendrobium officinale. The early developed 20 pairs of primers with good polymorphism were used for the analysis of 16 samples(15 samples of D.officinale and 1 sample of D.falconeri) by polyacrylamide gel electrophoresis, and the amplified bands were calculated. Nei's gene diversity and Simpson index were tested using PopGen32 to analysis the differentiation rate of single primer and combination primers. Finally, the construction of genetic similarity coefficient clustering map was calculated by using NTSYSpc2.1. The 16 pairs of primers, such as DN4, were highly polymorphic, while lower in DN13, DN23, DN60 and DN67. Although single primer could not distinguish all the samples, DN4, DN10, DN105, DN81, DN39 and DN71 displayed relatively higher distinguish rate. All the samples could be distinguished effectively by combined primers, and the DN4+DN10+DN105+DN39 primer set could be used as the core primer for fingerprint construction and genetic diversity analysis of D.officinale. By cluster analysis, the 20 pairs of primers could be used for the identification of both intraspecific and interspecific phylogenetic relationships of D.officinale. The primers DN13, DN39, DN4, DN58, DN65, DN67, DN10 and DN99 could be used for the identification of D.officinale and D.falconeri. The 20 pairs of primers could be wildly used in the application of genetic diversity analysis, germplasm identification, genetic relationship identification and genetic map construction of D.officinale.

Key words: Dendrobium officinale, molecular markers, EST-SSR, primer evaluation, applied research

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