DNA Barcoding of Highly Threatened Sacred Plant - Aegle marmelos L.

Abstract

DNA barcoding is a useful tool to define operational taxonomic units based on standardized DNA regions. In this study, the matK chloroplast gene was tested for species identification of highly threatened sacred plant Aegle marmelos L., to document their reliable identification, discrimination, similarities and evolutionary trend with their related taxa for future use. DNA isolation from leaf sample of Aegle marmelos was carried out by using a modified Cetyl Trimethyl Ammonium Bromide method (CTAB). Gradient PCR amplification was performed for the isolated DNA using matK gene and the primers, matK472F and matK1248R. Pairwise sequence alignments were made with BLAST and multiple sequence alignments were also made with ClustalW. Based on the sequence alignments, dendrograms were constructed using the software, Mega 5 and Neighbor Joining tree method was used to study the phylogenetic aspects of the species studied and with their related taxa. The success rate of PCR amplification was 90-95% and the success rate of DNA sequence was 95%. The maximum identity of the present study sample was 100% with Aegle marmelos and the similarity of other related species, Severinia buxifolia, Citrus trifoliata and Citrus reticulata was 97% respectively.