In vitro regeneration of Guizotia abyssinica Cass. and evaluation of genetic fidelity through RAPD markers

Abstract

The objective of this study was to induce a rapid as well as prolific shoot regeneration protocol for micropropagation and RAPD analysis of Guizotia abyssinica Cass. which is an important herbaceous plant of immense industrial value via direct and indirect organogenesis from apical bud, axillary bud, leaf and internode explants. Best seed germination was obtained on cotton irrigated with liquid MS medium. Out of the four explants used, apical bud proved to be the best in terms of shoot regeneration and multiplication. Best shoot multiplication was obtained from apical bud, axillary bud and leaf explants on MS medium supplemented with 2.22μM BAP+2.85μM IAA. Whereas supplementation of MS medium with 2.22μM BAP+28.55μM IAA produced maximum number of shoots from internode explants. BAP (0.44μM) in combination with Kn (0.46μM) proved suitable for maximum mean shoot length. Moreover, culturing the regenerated shoots on half-strength liquid MS medium supplemented with NAA (2.68μM) induced maximum rooting from elongated shoots (direct and indirect regeneration). The plantlets were established in plastic cups containing vermiculite, soil, sand and farm yard manure and then successfully transferred to field with 97.33% survival. Analysis of RAPD recognized 197 different amplification products and showed the presence of somaclonal variation in the plantlets arising from direct regeneration as well as from indirect regeneration. The protocol developed in this study is suitable for propagation of quality planting material for commercialization, germplasm conservation and for future genetic improvement studies.