DNA Barcoding of endangered Ceropegia odorata Nimmo ex J. Graham and Ceropegia hirsuta Wight & Arn. species of Gujarat

Abstract

In the present study, DNA Barcoding was made for the endangered plants Ceropegia odorata Nimmo ex J. Graham and Ceropegia hirsuta Wight & Arn. DNA barcoding is used to authenticate their reliable identification, conservation, discrimination, similarities and evolutionary trend among them and with their related taxa for future use. Two species, viz., Ceropegia odorata Nimmo ex J. Graham and Ceropegia hirsuta Wight & Arn are reported from Gujarat Dediyapada. The identification of both the species depends on the knowledge which held by taxonomists. Non-specialist cannot cover all the knowledge of the species. To handle with these difficulties’ DNA Barcoding aims to develop a standardized, rapid, and inexpensive species identification method approachable to non- specialists and will become easy for taxonomist. DNA isolation from leaf samples of present study species was carried out by applying a modified CTAB method and good isolation was just for the species considered. Gradient PCR amplification was performed for the isolated DNA using rbcL gene and the primers rbcLF and rbcLR were used. The amplification success rate was 90-95%. PCR amplification was tested with 1 % agarose gel electrophoresis using ethidium bromide and the products were confirmed. The PCR products were further processed for DNA sequencing and sequences were good 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 Neighbor joining method to study the phylogenetic aspects of the species studied and with their related taxa this study was to apply to support conservation efforts of Ceropegia Species in Gujarat and all over India. We used to recommend rbcL marker for both the sample. We specifically tested whether the markers could be used to solve taxonomic confusion concerning the Ceropegia species. This Barcoding system will be utilized specifically to identify and create phylogeny among the selected endangered species.