白及种子无菌萌发及组培快繁体系建立OA

With Bletilla striata seeds as explants,tissue culture seedlings were obtained through aseptic germination,pseudobulb induction,seedling boosting,and rooting culture.Experiments were conducted to investigate the effects of different phytohormone combinations on seed germination and pseudobulb induction,as well as the influences of different seedling types and substrate formulations on transplantation.A tissue culture and rapid propagation system for B.striata was established.The results showed that the optimal medium for B.striata seed germination was 1/2MS+GA3 0.1 mg/L+NAA 0.1 mg/L+sucrose 30 g/L.After 30 d of culture,the protocorm diameter reached 0.95 mm and the plant height reached 1.69 mm.The optimal medium for pseudobulb induction,seedling boosting,and rooting was 1/2MS+6-BA 0.8 mg/L+NAA 0.6 mg/L+PP333 0.3 mg/L+sucrose 30 g/L+activated carbon 0.1 g/L+banana puree 30 g/L.After 90 d of seedling boosting and rooting culture,the pseudobulb diameter reached 5.96 mm and the plant height reached 2.42 cm.The optimal transplantating method involved decapitating the tissue culture seedlings and using a mixed substrate composed of medicinal residue and garden soil at a ratio of 1∶1.After 90 d,the survival rate reached 95.3%,with new shoots exhibiting broad dark green leaves with vigorous growth.The established system directly induced pseudobulbs after the aseptic germination of B.striata seeds and enabled seedling boosting and rooting without the need for protocorm proliferation step,simplifying the operation,shortening the cycle,and reducing the risks of pollution and germplasm degradation.It provided technical support for the standardized and large-scale production of high-quality B.striata seedlings.