Studies On Embryogenesis And Micropropagation In Vitro Of Phyllostachys Violascens

Phyllostachys violascens is a superior edible bamboo species in China with wide market value. Recently the plants were undergoing declining and blooming in large area.In order to solve the problems in vitro tissue culture techniques by excising lateral buds and seed embryos as explants of Phyllostachys violascens thus were adopted for vegetative propagation. Experimental results showed as follows.1. Study micropropagation of Phyllostachys violascens via lateral bud culture in vitroThe new flushes of lateral buds were employed as explants. Experimental results demonstrated that MS salts+MT vitamins+Sucrose 30 g.L^-1+Gelrite 4 g.L^-1 was optimum basal medium for lateral shoot growth and multiplication. The basal medium supplemented with KT 1mg.L^-1 or 3 mg.L^-1 attained growth rats 81% and 72% on spring and autumn buds respectively after 30 days in culture. The basal medium supplemented with TDZ 0.03 mg.L^-1+KT 1 mg.L^-1 or IAA 0.03 mg.L^-1+2IP 1 mg.L^-1 attained proliferation rate 1.94 and 1.28 on spring and autumn buds respectively after 30 days in culture.2. Study micropropagation of Phyllostachys violascens via seed embryo cultures in vitroZygotic embryos were served as the explants. Experimental results indicated that the seeds were placed in running water for 1h,followed by disinfestation in NaClO 2.5% with tween 20 for 10 min in a vacuum. Rinse seeds with autoclaved water for 5-6 times. The 88% germination rate was achieved in MS medium (containing MS salts, MT vitamins, myo-inositol 100 mg L^-1, sucrose 30 g L^-1). Best proliferation was attained when supplemented with TDZ 0.01 mg.L^-1+BA 1 mg.L^-1+Banana Powder 2 g.L^-1.3. Study on in vitro propagation of adventitious bud formation and embryogensis via organogenic callus induction of Phyllostachys violascensZygotic embryos were served as the explants. Experimental results revealed MS salts with Adenine Sulfate 10 mg.L^-1, MALT Extract 0.5 g.L^-1, Glucose 20 g.L^-1 and Type A agar 10 g.L^-1 was selected as basal medium for inducting adventitious bud and embryogensis. Early early cotyledon were used as the best stage as explants for inducing compact callus formation in basal medium supplemented with BA 0.1 mg.L^-1 and Picloram 0.1 mg.L^-1. The rate 20.79% was attained on adventitious bud differentiation when compact callus was transferred to the basal medium supplemented with TDZ 0.01 mg.L^-1 and KT 0.3 mg.L^-1 after 2 weeks in culture. Embryoid formation or embryogenesis was up to 87.43% when embryogenic callus was transferred to the basic medium and supplemented with BA 0.1 mg.L^-1, TDZ 0.001 mg.L^-1, Picloram 0.1 mg.L^-1 and ABA 0.3 mg.L^-1 for 4 weeks, then removed ABA and added NAA 0.1 mg.L^-1 for 1 month. Small plantlets were finally observed when BA raised to 1 mg.L^-1 after 2 months in culture. Histological examinations on adventitious bud formation and embryogenic callus illustrated the big nuclei and dense cytoplasm cells appearance with organized structures.