The 18S rDNA gene discriminates between red-listed and unexplored ethnomedicinal species of Myristicaceae restricted to humid tropics of India

A potentially diagnostic 18S rDNA (ribosomal DNA) gene was amplified reliably from red-listed ethnomedicinal species of Myristica and its wild and related genera. Individuals from nine species of Myristicaceae were utilized for the study. The sequences ranged from 1,767 to 1,794 nucleotide (nt) in length. The GC content (%) varied from 52.77 to 51.04. The frequencies (%) of nt were A (23.31), T (23.82), C (24.48) and G (28.39). The alignment of all sequences produced 195/1,516 variable sites and 1,257/1,516 conserved sites. Total numbers of single nucleotide polymorphism (SNP) sites found in the alignment were 146/1,516. Knema andamanica (Warb.) W.J. de Wilde was the most distinct that included 18 variable regions and 15 InDel with 27 SNP sites, specific to this species. The identified regions from nine species of Myristica and its wild and closely related genera were deposited in the GenBank Database (Accession numbers JN228257-JN228265). Comparison of morphological identifications and phylogenetic analysis indicated that the specimens were correctly assigned on the basis of a short stretch of 18S rDNA (~1,600 bp) making this a potentially useful marker for the rapid molecular assignment of an unknown related species also. Significant sequence homology ranging from 72 to 99 % was observed on comparison with 18S rDNA genes of other plants in the public domain. A comparison of intraspecific data information of nine 18S with that of 73 matK and 86 rbcL sequences from GenBank revealed that polymorphism, divergence and conservation is higher in 18S locus for Myristicaceae. Hence these markers may be utilized for phylogenetic analysis, evaluation of species richness during ecological surveys or for environmental assessments. These molecular markers are especially important due to the fact that the species studied are mostly vulnerable and red-listed with limited availability in endangered ecological niches.