Assessment of Genetic relationship among Cannabis sativa L. in Thailand based on ISSR and their Phytoconstituents properties

Abstract

Due to the long history of prohibition, limited research on Cannabis sativa L., a Thai medicinal plant, remained unexplored for its scientific data to support their medicinal applications. The aim of this study was to assess their genetic relationship among C. sativa collected from different locations of Thailand as well as their phytoconstituents properties. Plant genomic DNA was isolated and genetic relationship was performed based on inter simple sequence repeat (ISSR). Seven out of twenty-five ISSR primers were amplified and yield 993 reproducible banding patterns ranking from 90 - 2,339 base pair in length, of which 99.3% were found to be polymorphic. The dendrogram by Unweighted Pair Group Method with Arithmetic Average (UPGMA) clustered into 2 main clusters. The phytoconstituents analysis of C. sativa ethanolic extract was performed according to the standard method and also their antioxidant property. Phenolic compounds, alkaloids, flavonoids, diterpenes, triterpenes and steroids were presented but not saponin. Total phenolic content assessed by Folin-Ciocalteu’s method was 14.08±0.34mg GAE/g extract while flavonoid content assessed by aluminium chloride was 23.88±0.40mg QE/g extract. Their antioxidant property was assessed using DPPH and FRAP assays expressed in term IC50 was found to be 112.13±0.09μg/ml and 0.228±0.12mM Fe2+/g extract, respectively. The obtained results reveal that ISSR markers could be useful for assessing of genetic relationship of Thai C. sativa collected from various areas and their phytoconstituents properties provided the useful information for their medicinal applications.