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Research articles

ScienceAsia 40 (2014): 113-120 |doi: 10.2306/scienceasia1513-1874.2014.40.113

Identification of native Dendrobium species in Thailand by PCR-RFLP of rDNA-ITS and chloroplast DNA

Surin Peyachoknagula,b,*, Chareerat Mongkolsiriwatanab,c, Sirinrat Wannapinpongd, Pattana Srifah Huehnea,e, Kornsorn Srikulnatha,b

ABSTRACT:     The PCR-restriction fragment length polymorphism (PCR-RFLP) approach was successfully developed to identify 25 native Dendrobium species in Thailand. PCR-RFLP of the rDNA-ITS with six restriction enzymes and three chloroplast (cp) DNA regions with five primer-enzyme combinations produced 24 types of DNA patterns altogether. Twenty-three out of the 25 species determined in this study were found to belong to unique classes and were successfully differentiated. Two species, D. crumenatum and D. formosum, possessing the same DNA pattern, however, were identified after cutting the chloroplast DNA fragment amplified by psbC-trnS primer with MboI enzyme. An effective procedure for identifying each Dendrobium species was developed. PCR-RFLP of the rDNA-ITS with TaqI, which is the most informative enzyme, was used for the early detection of 16 Dendrobium species. To identify the remaining Dendrobium species, PCR-RFLP analysis was performed using one more primer-enzyme combination. Our study provides a rapid, simple, and reliable identification method for these Dendrobium species.

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a Department of Genetics, Faculty of Science, Kasetsart University, Bangkok 10900 Thailand
b Centre for Advanced Studies in Tropical Natural Resources, National Research University-Kasetsart University, (CASTNAR, NRU-KU), Bangkok 10900 Thailand
c Department of Genetics, Faculty of Liberal Arts and Science, Kasetsart University, Nakhon Pathom 73140 Thailand
d Institute of Food Research and Product Development, Kasetsart University, Bangkok 10900 Thailand
e Biotechnology Laboratory, Chulabhorn Research Institute, Bangkok 10210 Thailand

* Corresponding author, E-mail: fscisrp@ku.ac.th

Received 12 Mar 2013, Accepted 10 Mar 2014