RAPD markers for genetic analysis and classification of Musa B genome

Abstract

Random amplified polymorphic deoxyribonucleic acid (RAPD) markers were determined to classify the B genome of Musa germplasm collections. Optimization of several parameters for DNA amplification and RAPD analysis was also done to determine RAPD reaction conditions applicable for Musa cultivars in the B genome. A reaction mixture of 35 ng DNA, 2.5 mM MgCl2,1 unit/μL Taq DNA pol, 1μM dNTPs, 0.50 ng RAPD primer and 1X PCR buffer was found optimum for DNA amplification. The RAPD polymerase chain reaction (PCR) temperature cycle profile developed for rice was similarly found applicable for the Musa cultivars evaluated. Moreover, a random ten-mer primer with 60%-70% G+C obtained good RAPD band pattern. Specifically, primers OPH-03 and OPI-16 detected three DNA markers which were present in all BB/BBB cuttivars. These DNA markers (0.560 kb marker detected by primer OPH-03 and 0.836 and 0.690 kb markers detected by primer OPI-16) were therefore used to determine the genome constitution of three Pinatubo collections and Palawan IX. Cluster analysis using the RAPD markers data showed that the balbisiana cultivars are different from acuminata cultivars as represented by 'Bungulan' and 'Lakatan', respectively. Furthermore, the Pinatubo collections were shown to be of the balbisiana group. However, the genome classification of the Palawan collections was not clear because of inconsistencies in clustering.

Source or Periodical Title

Philippine Agricultural Scientist

ISSN

317454

Page

165-173

Document Type

Article

Subject

B genome, Markers, Musa, Primers, RAPD

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