Callus Culture From Bagged Seedlings Leaves of Hevea Brasiliensis Müll. Arg. Clone Reyan 7-33-97
In order to expand the rubber tree explant sources and to establish efficient and stable rubber tree leaves in vitro regeneration system, the annual bagged seedling leaves of Reyan 7-33-97 were used as the explants to study on the effects of leaves pretreatment, different disinfection time with 0.1% HgCl2, different parts of detached leaves, different combinations of three kinds of phytohormone(6-BA, NAA, 2,4-D) on callus induction frequency, and screen the best subculture medium. The results showed that leaves pretreated with liquid medium MS+3 mg/L 6-BA+0.5 mg/L 2,4-D+0.2 mg/L NAA could induce callus; leaves disinfected with 0.1% HgCl2for 15 min was the best disinfection method, its survival ratio could be 2.47%; leaves(with principal vein) and no callus part of leaves(with principal vein) already produced callus were the most suitable explants for rubber bagged seedlings leaves, callus induced by which showed tighten and yellow-green; the optimum callus induction medium was MS+3 mg/L 6-BA+0.5 mg/L 2,4-D+0.2 mg/L NAA+4% sucrose+3 g/L phytagel, leaves(with principal vein) and no callus part of leaves(with principal vein) have already produced callus were cultured in this medium, their callus inducing ratio can amount to 49.07% and 42.22%. Conclusion: after pretreatment, choose leaves(with principal vein) of annual bagged seedling leaves of Reyan 7-33-97 as explants that can be used to induce ideal callus.