Studies on DNA barcoding of sacred plant - Ficus religiosa L.

Abstract

In the present investigation, DNA barcoding were made for the sacred trees of Main (Big) Temples in Cuddalore district, namely Ficus religiosa, to document their reliable identification, discrimination, similarities and evolutionary trend among them and with their related taxa for future use.  DNA isolation from leaf samples of present study species was carried out by using a modified CTAB method and good isolation was got for the species studied. Gradient PCR amplification was performed for the isolated DNA using matK gene and the primers matK472F & matK1248R.  The amplification success was 90-95%. PCR amplification was tested with 1 % agarose gel electrophoresis using ethidium bromide and the products were confirmed.  The PCR products were sent to Xcelris Labs Ltd. Sydney House, Premchand Nagar Road, Bodakdev, Ahmedabad 380054, India, for DNA sequencing and sequences were got for the species with the success rate of 95 %.  Pairwise sequence alignments were made with BLAST and multiple sequence alignments are made with ClustalW, and based on the sequence alignments, dentrograms were constructed using software Mega 5 and Neighbor joining method to study the phylogenetic aspects of the species studied and with their related taxa.