铁皮石斛无菌播种产业化繁育技术研究

Study on technology of aseptic sowing and rapid propagation of Demdrobium officinale

关键词:  铁皮石斛  无菌播种  原球茎  产业化

Key words:  Demdrobium officinale  aseptic sowing  protocorm  industrialization

摘要:

以铁皮石斛的蒴果为外植体，采用种子→原球茎→完整植株→移栽的途径快速成苗进行工厂化生产，对各阶段培养基进行筛选，以及其他一些影响因子进行比较研究。结果表明：人工授粉后生长60～180 d的铁皮石斛种子在离体条件下均能萌发，其中授粉150～180 d种子的萌发效果最好，萌发率为87.2%～944%，适宜的萌发培养基为MS+6 BA 1.0 mg/L+NAA 0.1 mg/L+马铃薯汁200 g/L+AC 1.0 g/L；原球茎增殖的最佳培养基为MS+6 BA 1.5 mg/L+NAA 0.1 mg/L+香蕉汁100 g/L+AC 1.0 g/L，繁殖系数约为20倍/50 d；原球茎在MS+6 BA 1.0 mg/L+NAA 0.1 mg/L+马铃薯汁200 g/L+AC 1.0 g/L培养基上进行分化培养，分化的同时还能进行一定的增殖；将已分化的芽苗转接到壮苗培养MS+6 BA 0.5 mg/L+NAA 0.2 mg/L+香蕉汁100 g/L+AC 1.0 g/L上培养1代后，转接到生根培养基1/2MS+NAA 0.8 mg/L+无机盐A 0.2～0.5 mg/L +香蕉汁100 g/L+AC 1.0 g/L上，培养50～70 d后，生根率100%，无机盐A可以有效地控制愈伤或原球茎的形成，明显提高生根苗的数量和质量。在桂林地区，生根苗以3～5月和9～10为最佳移栽期，以通过高温处理并堆沤腐熟的松树皮为基质，移栽成活率可达90%。

Abstract:

Seeds of Demdrobium officinale could be used as explants to rapid propagate by the way of seed→protocorm→the whole plant→transplant，and medium in each culture stage and some other factors were studied comparatively. The results showed that all seeds of D.officinale which were 60-180 d，especially 150-180 d after pollination could germinate，with a germination rate of 87.2%-94.4%. The suitable medium for seed germination was MS+6 BA 1.0 mg/L+NAA 0.1 mg/L+potato 200 g/L+active carbon 1.0 g/L. The best medium for the formation and multiplication of protocorm was MS+6 BA 1.5 mg/L+NAA 0.1 mg/L+banana mud 100 g/L+active carbon 1.0 g/L，the propagation coefficient was about 20 times/50 d. And the protocorm differentiated on MS+6 BA 1.0 mg/L+NAA 01 mg/L+potato 200 g/L+active carbon 1.0 g/L medium，which could have some multiplication in the meantime. After being cultured on MS+6 BA 0.5 mg/L+NAA 0.2 mg/L+banana mud 100 g/L+active carbon 1.0 g/L for a generation，the vigorous plantlets were transferred to rooting medium which was 1/2MS +NAA 0.8 mg/L+mineral salt A 02-0.5 mg/L +banana mud 100 g/L+active carbon 1.0 g/L and were cultured for 50-70 d，with 100% rooting rate. Mineral salt A could effectively control the formation of callus or protocorm，and obviously improve the quality and quantity of root regeneration plants. The best time for the transplantation of root regeneration plants was March May and September October in Guilin area. The plantlets were transplanted into rotten pine bark handled with high temperature in seedbed of greenhouse resulted in more than 90%survival rates.