DNA barcoding of Lamiaceae from Poonch valley, Azad Jammu and Kashmir, Pakistan

Lamiaceae is the sixth-largest angiosperm family and the largest one in the order 'Lamiales'.The family members are widely used for medicinal purposes and to produce perfumes, oils, and culinary products.There are 21 Genera and approximately 36 species of Lamiaceae in Poonch valley, Azad Jammu and Kashmir (AJK), Pakistan.It is important to identify the Lamiaceae species to explore its taxonomical status.In current research DNA barcoding of Lamiaceae was performed to ensure correct identification of Lamiaceae species from Poonch valley, Azad Jammu and Kashmir (AJK), Pakistan.For this purpose, Lamiaceae plants were collected from different regions of Poonch valley, high quality DNA was extracted and subjected to amplification by three plastid barcoding loci (rps11, rbcLa, and trnL-trnF).The amplified products were purified and sequenced.Electropherograms were assembled and edited by Geneious 6.1.8.The novel sequences were compared against previously available data using the Basic Local Alignment Search Tool (BLAST).The consensus sequence lengths of the rps11, rbcLa, and trnL-trnF loci in the studied Lamiaceae species were 417, 573, and 375 base pairs, respectively.The overall PCR amplification success rates were 97% for rps11, 85% for rbcLa, and 75% for trnL-trnF.The obtained sequences were validated against the BOLD and GenBank databases for species identification.Nucleotide BLAST analysis revealed 73% sequence similarity with the corresponding species, whereas 73-100% with the corresponding genera.In our study comparatively rbcLa provided high species-level discrimination (73%) and proposed as the most effective DNA barcode among the three evaluated loci for species-level identification of Lamiaceae medicinal plants.To investigate the phylogenetic relationships of the studied Lamiaceae species, phylogenetic tree was constructed using the obtained DNA barcode sequences.The phylogenetic tree improves the placement of species in clades.This study would be useful to understand the phylogenetic relationships and confirming the taxonomic identity of medicinal plant species within the family Lamiaceae.