Intraspecific variation of bitter gourd Momordica charantia Linn. by RAPD analysis

Abstract

Ramdomly amplified polymorphic DNA (RAPD) analysis was used to determine the levels of genetic diversity and to estimate genetic relationship among 22 accessions of Momordica charantis Linn. (Mara-Khee nok) from Pichitr Horticultural Research Center, Thailand. Six of 47 sceened decanucleotide primers (X-12, S-16, X-18, X-24, X-26 and X-27) gave reproducible RAPD patterns. One hundred and nine reproducible RAPD bands were generated across the investigated species of these, 54 bands (49.5%) were monomorphic band and 55 bands (50.5%) were polymorphic band. Genetic variations of 22 accessions of M. charatia were analyzed by neighbor-joining program and the dendrogram divided Mara-Khee nok into 2 groups that were correlated diameter size of the fruits. However, the relationship between the division of Mara-Khee nok and their geological distribution was not indicated. The expression of MAP30 gene was investigated in apical shoot, leaf, flower bud, male flower, female flower, young fruit tissue, seed of young fruit, rie fruit tissue and seed of ripe fruit. The MAP30 gene expression of apical shoot, leaf and seed of young fruit were found. The amplification of MAP30 gene from genemic DNA template and mRNA template gave PCR product of equal size. Cloning of MAP30 gene into plasmid pUC18 for expression in Escherichia coli was not successful. The MAP30 gene fragment was not ligated to plasmid pUC18. It might be caused from wrong designing at the 5'-end restriction site of specific primer for amplification of MAP30 gene before cloning.