Tissue Culture And Cultivation In Haplocladium Microphyllum

By field inspection and the comprehensive analysis, the growth conditions for Haplocladium microphyllum tissue culture and soil cultivation were suggested, that is to say, the illumination intensity was 1500-3000lx, the moisture was 50-90%, and pH was 5.96-8.00. By means of spore culture, protonema culture and gametophyte culture, the development process of H. microphyllum were discussed, and fast reproductive system of H. microphyllum were established.The result showed that the spore was easy to sterilize, after washed the spore in 70% alcohol for 6 min, sterilized the spore in 20% NaClO for 5 min the contamination rate was 0. By orthogonal test design for the screening of the disinfection methods for the gametophyte, the results showed that the best way of the gametophyte sterilization was as followed: Explants gotten from intersection of branch were washed in 70% alcohol for 5s, and then washed in 2.5%NaClO for 60s, the survival rate was 70%.By means of sporeâ†'protonemaâ†'moss plant, sporeâ†'protonemaâ†'callusâ†'moss plant, gametophyteâ†'callus/budâ†'moss plant, the results showed that the optimum medium was MS+30g?L-1 sugar + 6.7 g?L-1 agar+pH 7.0. Added 1mg?L-1IBA in the medium for the protonema could get the most number of moss plant, and the moss clump was the highest, the color of the protonema was deep green, the comsistency of protonema was compact. Added 0.5 mg?L-1KT in the medium for the gametophyte was best to the induction and growth of bud. Added 0.3 mg?L-16-BA is best for the callus induction. Added 1mg?L-1IBA in the MS medium is best for the callus differentiation.Effects of different soils on growth and development of H. microphyllum was studied, and the soils which used to cultivated H. microphyllum were yellow upland soil, yellow sand soil and white clay. The experiment was been tested in illumination incubator. Cover degree, branch length, branch number, biomass, relative moisture content and photosynthetic pigment content had been evaluated on growth and development of H. microphyllum. The study showed that yellow upland soil was the best for growth and development of H.microphyllum, 50ds later after cultivated in yellow upland soil, H. microphyllum cover degree reached 84.33%, branch length was 4.80cm, level 1 branch number was 11.87 and level 2 was 5.13, fresh weight and dry weight increased 38.15 times and 17.69 times respectively.