Study On The Shallow Liquid Tissue Culture And Polyploid Induction In Amorphophallus

This experiment studied on the shallow liquid tissue culture in Konjac and Amorphophallus Bulbifer, and Konjac polyploid induction by combining with shallow liquid tissue culture and colchicine solution soaking method.The results showed:A. Konjac tissue shallow liquid(5mm, lOmL) tissue culture system was established by this experiment. The result showed that shallow liquid medium was prior to solid medium in the browning control, callus growth speed, number of differentiation buds, robustness of roots and cost. MS+6-BA0.5+NAA0.5+Sucrose3.0%+0.1%PVP (pH5.8) could be used for primary culture. Alternative culture in MS+6-BA2.0+NAA0.5+Sucrose3.0%+0.1%PVP (pH5.8) and MS+ 6-BA1.0+NAA0.5+Sucrose3.0%+0.1%PVP (pH5.8) could produce tissue culture plantlets largely and continuously. Rooting medium could be 1/2MS+IAA0.5+Sucrosel.5%+0.1%PVP (pH5.8). Besides, three types of callus were found after subculture for three times. Remarkable difference existed in the A, B and C types in appearance, organization structure and capacities of redifferentiation. A and B-type callus were chosen for bud differentiation, in both test and application, which could produce a stable and efficient differentiation system.Amorphophallus bulbifer shallow liquid(5mm,10mL) tissue culture was established. The result showed that MS+6-BA 0.5+NAA 0.5+Sucrose 3.0%+0.1% PVP (pH5.8) could be used for primary culture. Alternative culture in MS+6-BA 2.0+NAA 0.5+Sucrose 3.0%+0.1% PVP (pH5.8) and MS+6-BA 1.0+NAA 0.5+Sucrose 3.0%+0.1% PVP (pH5.8) could produce tissue culture plantlets largely and continuously. Rooting medium could be 1/2MS+IAA 0.5+Sucrose 1.5%+0.1% PVP(pH5.8).The results showed that shallow liquid tissue culture are better than solid culture medium on primary culture browning rate, callus induction, bud polarization, root polarization and cost.Basing on the shallow liquid tissue culture system, this experiment studied Konjac polyploid induction. The experiment conditions as follows:The concentration of colchicine solution (with DMSO 2%) is 0.005%,0.01%,0.05% and 0.1%; the processing temperature is 15℃. The results showed that the survival and the polarization rate of explants are declined, with concentration of colchicine solution and treatment time is improved, but the mutation rate rose at beginning then declined simultaneously. When we soak the experiment callus with colchicine solution 0.1% and 2 hours in 15℃, the survival rate is 83.3%, the polarization rate is 64.0% and the mutation rate is 41.53%.The chromosome number of root tip showed that the mutation plant is tetraploid. This experiment also studied on the morphology features. The results showed that the tetraploid plant is brachysm, leaf became wider and leaf index became smaller. Those results fully embody the organs is huge. And Konjac tetraploid plants'leaf is more deep green. The stoma of lower epidermis and the guard cells is magnified, but the stomatal frequency is decreased.