GENETIC ANALYSIS ON HIBISCUS SPECIES BY USING RAPD MARKERS

Abstract

This study was undertaken for genetic analysis within the 7 varieties of Hibiscus rosa-sinensis by using RAPD markers the method involves a extraction of DNA from leaves of Hibiscus rosa-sinensis and yield of DNA ranged from 158-200µg and purity (ratio) was between 1.3-1.6 indicating minimum level of contaminating metabolites. Genetic analysis and relationship among seven Hibiscus rosa-sinensis germplasm were analyzed using Random Amplified Polymorphic DNA (RAPD). Total nine primers were used as, RPI-07, RPI-08, RPI-10, RPI-12, RPI-18, RPI-19, RPI-21, RPI-22 and RPI-23. No PCR product was generated through the primer RPI-23 indicating that no binding sites for this primer. In all the primers RPI-07, RPI-08, RPI-10, RPI-19 and RPI-21 are more suitable for the genetic variation analysis in Hibiscus species as it has shown maximum polymorphic bands. Primers RPI-12, RPI-18 and RPI-22 are weak in showing the polymorphic banding pattern for the all species of Hibiscus rosa-sinensis. By using PAST and MEGA programme Consensus Phylogenetic tree generated based on genetic distances segregated the seven Hibiscus germplasm into two different clusters. 75% times of species Hibiscus rosa-sinensis Red S.P. and D.P. (Single petal, Scarlet and Double petal, Rubroplenus) clustered in a single cluster. 87% times the cluster of Hibiscus rosa-sinensis Pink varieties clustered with Hibiscus rosa-sinensis White varieties. The results of the present study indicated that the RAPD analysis could be utilized by breeders for further improvement of Hibiscus rosa-sinensis species.