Phylogenetic relationship was studied among three species of rubber available in India, Hevea brasiliensis, H. benthamiana and H. spruceana by employing different molecular marker techniques namely, RAPD, chloroplast DNA (cpDNA) PCR-RFLP and heterologous chloroplast microsatellites. RAPD analysis clearly indicated a high degree of polymorphism among the three species. Using twenty-five arbitrary primers, a total of 305 bands were amplified in the clones RRII 105 and GT 1 belonging to H. brasiliensis, H. bentbamiana, H spruceana and FX 516, an interspecific hybrid of H. benthamiana and H. brasiliensis, out of which 256 bands were polymorphic. Analysis of the interrelationships among the species clearly revealed that the clones of H. brasiliensis (>50% genetic dissimilarity) are closer to H. benthamiana than to H. spruceana (>70% genetic dissimilarity). Species-specific RAPD markers were identified for each species and their locus specificity was proved through hybridization. RFLP analysis using three hyper variable intergenic spacers, rbcL-ORF106, trnM-rbcL and trnC-trnD could not detect variability. Among different primer-pairs for heterologous chloroplast microsatellites tested, five could successfully be amplified, of which one (ccmp6) was highly polymorphic and could detect both intra- and inter-species polymorphism to prove the maternal mode of inheritance of the chloroplast genome in Hevea.
Chloroplast microsatellite, cpDNA polymorphism, DNA marker, Hevea brasiliensis, Hevea benthamiana, Hevea spruceana, Phylogenetic relationship, RAPD