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

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60～180d的铁皮石斛种子在离体条件下均能萌发,其中授粉150～180d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 1.0mg/L+NAA 0.1mg/L+马铃薯汁200g/L+AC 1.0g/L;原球茎增殖的最佳培养基为MS+6-BA 1.5mg/L+NAA 0.1mg/L+香蕉汁100g/L+AC 1.0g/L,繁殖系数约为20倍/50d;原球茎在MS+6-BA 1.0mg/L+NAA 0.1mg/L+马铃薯汁200g/L+AC 1.0g/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 0.5mg/L+NAA 0.2mg/L+香蕉汁100g/L+AC 1.0g/L上培养1代后,转接到生根培养基1/2MS+NAA 0.8mg/L+无机盐A 0.2～0.5mg/L+香蕉汁100g/L+AC 1.0g/L上,培养50～70d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%。

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.