Study On Tissue Culture And Cell Suspension Culture Of Stylosanthes

This paper studies comprehensively on the induction of aseptic seedings, callus induction, multiplication and differentiation from cotyledon and hypocotyls, rooting and transplanting of tissue culture shoots, stem culture, anther culture, and the cell suspension culture etc. of Stylosanthes.The preliminary experiments' results as follows:1. Explants take place when they are cultured with NAA content 2.0mg/L, 0.1-0.3mg/L 6-BA can stimulate dedifferentiation, callus formation and bud regeneration.2. With proper 6-BA, NAA can improve callus quality, accelerate its growth rate, and stimulate bud formation. Higher NAA combined with proper 6-BA can increase production of bunch buds. Interaction of NAA and 6-BA is affected by different illumination and temperature conditions.3. Callus initiation of explants pretreated by low-temperature-short photoperiod is earlier than that of non-temperature-short-photoperiod or diurnal-temperature-difference.4. The optimum recipe of callus induction of hypocotyls and cotyledon is MS+0.4 mg/L 6-BA+2.0 mg/L NAA. The optimum recipe of differentiation from hypocotyls and cotyledon's callus is MS+2.0 mg/L 6-BA+0.25 mg/L NAA. The best rooting medium is 1/2MS+1.0 mg/L IBA+0.5 mg/L IAA.5. The stem of seedlings can immediately induct bud, the best material is the stem with axillary bud, of which the optimum recipe is MS+6-BA 5.0 mg/L+GA 0.5 mg/L,the inductivity was 90%; when the subculture medium of test-tube plantlets was MS+6-BA 1.0 mg/L+NAA 0.5 mg/L, the percentage could reach 100%, the rooting medium of test-tube plantlets was MS+IBA 1.0c+IAA 1.0 mg/L.6. The anther were treated by low temperature pretreatment for 24 hours was better than other treaments. The best medium for induction of cullus is N6+medium supplemented with 2,4-D(0.5 mg/L), KT(1.0 mg/L) was the best for induction of cullus.7. The suitable culture condition for Stylosanthes'cell suspension culture are following:the basic medium is MS, the hormone combination is NAA 2.0 mg/L +6-BA 0.3 mg/L, the initial inoculation concentration is 1.0 g/50mL, the pH value is 5.8, and the sucrose concentration is 40g/L.