Studies On Micropropagation In Vitro Of Phyllostachys Pubescens Embryo

Excised embryos of a series Phyllostachys pubescens were served as explants, and preliminary tissue culture system for micropropagation was established through primary culture, subculture and transplant following the hardening. In primary culture, MS basic medium was applied on embryo culture dissect, surface disinfestations, dark period and various kind and concentrations of cytokines, carbon sources tested embryo-derived micropropagation. Factorial tests were designed to perform experiments in order to detect the suitable conditions on cytokines, vitamins, and organic addenda. Selected conditions were established after concentrations of sugars, macro salts of MS salts concentrations and auxins for rooting were experimented, followed by hardening and transplanting. Results of each step were carried out with statistical analyses. Experimental results showed that:1. Seeds required pre-soaking or in running water in top water for 12¹6 h, 4 weeks later, the seed germination with soaking 12 h was in excess of 90%. Disinfestations of seed coat was done in 0.25% sodium hypochlorite solution with a few drops of Tween20 for 5 min in a vacuum would lower contamination rate less than 10%, and germination rate was 90%. BA and TDZ were prerequisite on the induction of embryo derived shoot multiplication, the concentration of BA between 0.3³.0 mg·L^-1, or TDZ between 0.001⁰.1 mg·L^-1 was better for growth. Sucrose played an important role on enhancing shoot growth and the browning degree, 4 weeks later, shoot with 30 g·L-1 sucrose grew well.2. Factorial tests indicated the combinations of BA and TDZ displayed significant affection than alone, BA0.3 mg·L^-1+TDZ0.1 mg·L^-1 hastened multiplication. Adding 0.01 mg·L^-1Picloram hastened growth of plants, the subculture medium was BA0.3 mg·L^-1+TDZ0.1 mg·L^-1+Picloram 0.01 mg·L^-1+sucrose 30 g·L-1+Gelrite 4 g·L-1, growth coefficient was 2.2, shoot grew vigorously. Adding MT or White's was not influence on multiplication of plants. Treatment with no Organic addenda was the best. Cultured in turn with MS medium and subculture would be favorable for multiplication, growth coefficient was 2.7, and average shoot length was 15.0 mm.3. Medium with sucrose 20 g·L-1 or 3/4MS was suitable for rooting. After the shoots regenerated, the MS medium with lower NAA or IBA resulted rooting of shoot. Transplanted in the artificial mixture with the ratio of peat: vermiculite: perlite = 1:1:1 was suitable; the rate of survival was 80%.