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논문검색

Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

초록

영어

In Malaysia, Labisia pumila Benth & Hook f, popularly known as ‘Kacip Fatimah’ has been used traditionally to treat various elements of the woman’s health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-40 °C were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHOD
 RESULTS AND DISCUSSION
 CONCLUSIONS
 ACKNOWLEDGEMENTS
 LIETERATURE CITED

저자정보

  • Subhash J Bhore Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Bedong-Semeling Road, Bedong, 08100,Kedah Darul Aman, Malaysia
  • A H Nurul Molecular Biology Division, Melaka Institute of Biotechnology, Lot 7, Melaka International Trade Center City, Ayer Keroh, 75450, Melaka, Malaysia
  • Farida H Shah Novel Plants Sdn. Bhd., Taman Rambai Harmoni, Off Jalan Tanjuag Minyak, 75260, Melaka, Malaysia

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