DNA Profiling of Biotypes of Trichosanthes Tricuspidata; an Important Ethno medicinal Cucurbit Using RAPD, ISSR and Scot Molecular Marke

Abstract

Trichosanthes tricuspidata Lour., is an important ethnomedicinal plant belonging to the Cucurbitaceae family. It is known by various vernacular names like Red ball snake gourd in English, Lal Indrayan in Hindi, Kalayar in Malayalam, and Avaduta in Telugu. There are several studies about the medicinal applications of Trichosanthes tricuspidata, but there is scanty information about molecular level identification of different biotypes of Trichosanthes tricuspidata. Hence, the present study was undertaken to develop a protocol for genetic analysis of three biotypes of Trichosanthes tricuspidata collected from wild areas of Warangal, Karimnagar and Adilabad districts of Telangana State, India by using Randomly Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Start Codon Targeted (SCoT) primers-based PCR method. Genomic DNA was isolated from 1-gram fresh leaf material of three biotypes of T. tricuspidata. The primers OPA-2 of RAPD, S-5 of ISSR and S-8 of SCoT has shown good amplification with generation of monomorphic DNA bands ranging from 250 bp to 1000 bp by OPA-2 of RAPD primer; 500 bp to 1500 bp by S-5 of ISSR primer and 1450 bp by S-8 of SCoT primer. In conclusion, no genetic variation was detected in three biotypes, as the generated DNA bands from OPA-2 of RAPD, S-5 of ISSR and S-8 of SCoT primer-based amplifications were monomorphic. The developed protocol of genetic analysis of three biotypes of T. tricuspidata by RAPD, ISSR and SCoT primers could be used in near future for molecular level studies.