African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12269

Full Length Research Paper

A comparison of nrDNA internal transcribed spacer (ITS) loci for phylogenetic inference and authentication among Cinnamomum osmophloeum and related species in Taiwan

Sahar H. Ahmed
  • Sahar H. Ahmed
  • Laboratory Technology Department, Faculty of Applied Medical Science, Misr University for Science and Technology, Egypt.
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Dina Sabry
  • Dina Sabry
  • Medical Biochemistry and Molecular biology, Faculty of medicine Cairo University, Cairo, Egypt.
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Olfat Noh
  • Olfat Noh
  • Obstetric and Gynacology, Faculty of Medicine Cairo University, Cairo, Egypt.
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Mai Samir
  • Mai Samir
  • Medical Biochemistry and Molecular biology, Faculty of medicine Cairo University, Cairo, Egypt.
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Kuen-Yih HO*
  • Kuen-Yih HO*
  • Department of Forestry and Nature Resources, National Chiayi University, Chiayi 60004, Taiwan.
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Shih-Chieh LEE
  • Shih-Chieh LEE
  • Department of BioIndustry Technology, Da-Yeh University, Changhua 51591, Taiwan.
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Shr-Wei SHIUE
  • Shr-Wei SHIUE
  • Department of Forestry and Nature Resources, National Chiayi University, Chiayi 60004, Taiwan.
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Chung-Chi HWANG
  • Chung-Chi HWANG
  • Department of Bioresources, Da-Yeh University, Changhua 51591, Taiwan.
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  •  Received: 26 December 2014
  •  Accepted: 02 March 2015
  •  Published: 01 April 2015

Abstract

Cinnamomum osmophloeum, an indigenous species of Taiwan, can be utilized for valuable products such as a food, a spice and a traditional Chinese medicine. This study compares the ribosomal DNA (nr DNA) internal transcribed spacer (ITS) sequence of C. osmophloeum to that of several other species with similar external morphology, such as Cinnamomum burmannii, Cinnamomum insularimontanum, Cinnamomum macrostemon and Cinnamomum subavenium. Phylogeny of ITS sequences shows that C. osmophloeum is more closely related to C. burmannii than the other species, while C. insularimontanum, C. macrostemon, and C. subavenium are phylogenetically relevant to each other. By comparing ITS sequence between C. osmophloeum and C. burmannii, specific primers were designed for the multiplex-PCR to differentiate them. Based on ITS sequence differences, all tested Cinnamomum spp. can be properly authenticated. A 125 bp band specific for C. osmophloeum and a 204-bp C. burmannii-specific band were successfully amplified by polymerase chain reaction (PCR) using the respective primers described above. The two species then can be identified at the molecular level according to the sizes of their respective PCR products as determined by gel electrophoresis.
 
Key words: Cinnamomum osmophloeum, internal transcribed spacer (ITS), phylogenetic, multiplex- polymerase chain reaction (PCR).
 

 

Abbreviation

Abbreviations: nr DNA, Ribosomal DNA; ITS, internal transcribed spacer; CTAB, cetyl trimethyl ammonium bromide; PCR, polymerase chain reaction; NCBI, National Center for Biotechnology Information; NJ, neighbor-joining method; MP, maximum parsimony; BLAST, basic alignment search tool.