Detection of somaclonal variation in micropropagated Hibiscus sabdariffa L. using RAPD markers

Abstract

The main objective of micropropagation is to produce clones i.e. plants which are
phenotypically and genetically identical to the mother plants. The culture of organized meristems usually guarantees the production of true-to-type plants but variations in the progenies have been widely reported. Hibiscus sabdariffa L. plants were regenerated on MS (Murashige and Skoog) medium containing BAP (Benzyl amino purine) and IBA (Indole 3 butyric acid) and were propagated in vitro on hormone-free MS medium. The aim of this study was to detect variation in micropropagated plantlets of Hibiscus sabdariffa using RAPD amplification. DNA extraction from Hibiscus sabdariffa L. plants was optimized using CTAB buffer supplemented with 5M NaCl to eliminate polysaccharides and the isolated DNA proved amenable to PCR amplification. RAPD analysis was carried out on DNA samples to compare the mother plant with 10 randomly selected regenerated plants. Out of 30 primers screened, primers OPB-01, OPX-06 and DK-02 produced polymorphic bands. These results show that RAPD is a suitable technique which can be used to detect genetic change caused by somaclonal variation and could be promising for the selection of desirable traits or transformation systems.

Keywords: Hibiscus sabdariffa L. In vitro culture. RAPD, Somaclonal
Variation