Establishment Of An In Vitro Regeneration System Of Tortula Ruralis

Tortula ruralis provided important ecological application value due to the endurance ability of drought, and its ecological function should not be ignored. Using tissue culture method, the regeneration system of T. ruralis was successfully built with new-born stem tips as explants. In the process, we investigated the effects of medium types, temperature, light, pH, agar concentration, sucrose concentration, plant hormone types and concentration on the growth of T. ruralis and expand reproduce. The results are as follows:1. The in vitro culture of the moss T. ruralis was established from young stem tips, and both protonema and gametophyte stages development were obtained after 90 days. Because of the serious pollution of wild materials, we recultivated the material to obtain newborn part with less pollution and strong vitality so that the material can be disinfected effectively. The optimal disinfection method is under 3% NaClO disinfection 5 min, and sterile water flushing, then 0.05% HgCl2 disinfection 20 s again, steriled water flushing 6~7 times.2. Beneck medium was suitable for disinfection?protonema extension and gametophyte development. MS medium was not suitable for disinfection. The material was transferred to MS?1/2 MS and 1/10 MS after disinfection, protonema was not found and the gametophyte differentiation appeared directly. The material died after transferring onto Knop medium.3. The T. ruralis was growing well on neutral partial acid medium; the T. ruralis growth speed and agar content showed a negative correlation; the T. ruralis was growingbest when the temperature was 20?, the best light intensity was 4 000 lx and the best illumination time was 12 h/d; the sucrose concentrations tended to have a positive effect on protonema formation and elongation, with the concentration increasing, the effect was more obvious, but high concentration sucrose inhibited buds differentiation.4. IAA and IBA significantly promoted stem elongation growth; 2, 4-D also obviously promoted the T. ruralis to grow, but at the same time accelerated aging; 6-BA prompted stem tip swelled into a ball; KT could induce the loose and black callus, and the gametophyte successfully developed after callus was transferred onto culture medium.5. The rhizoids?leaves and stem segments of T. ruralis were transferred to Beneck medium respectively, and the rhizoid proliferation was the greatest, the leaf was the second and stems proliferation was the least. Compared with the control the biomass of the segmentation of plants increased far beyond, the biomass could be up to 20 times about 90 days. Steriled gametophyte after grinding was transferred to the steriled culture medium. And the protonema was growing well and some protonema differentiated into gametophyte.