Establishment Of Tissue Culture System For Puccinellia Tenuiflora

In this paper, Puccinellia tenuiflora was selected as material. Tissue culture system was developed for plant regeneration from mature seed of Puccinellia tenuiflora. We have selected the most suitable conditions for the callus growth according to the effect of many factors, such as the different processing and disinfection of seed, basic medium, the compounding proportions of different hormones, sugar concentration and pH. And then the good embryo callus will be take differentiation and root.The main results are as follows:(1) The tissue culture system of Puccinellia tenuiflora was developed from mature seed, seed germination rate can improve greatly after peeling, so as to increase the callus induction rate obviously.And the callus induction rate from seed is higher than the one from young spikelet, stem and leaf.(2) The best seed disinfection time is3min of75%alcohol and10min of50%NaClO.(3) Compared with the B5, MS is more suitable for the basic medium of tissue culture of Puccinellia tenuiflora and the callus hydration rate reduced significantly.(4) The induction rates of primary callus from mature seed of Puccinellia tenuiflora were up to55.2%on callus induction medium contained MS basal salt and vitamins,5.0mgL-12,4-D and3%sucrose and solidified with0.7%agar, respectively. The little white callus can be seeing after2weeks,2weeks earlier than inducing by stem and leaf.(5) The optimal pH of the callus induction is7-8. Hydration rate decrease by half when Ph is from5.8to7-8.(6) The differentiation rates of embryo callus were31.7%on differentiation medium contained MS basal salts and vitamins,1.Omg·L-16-BA and0.5mg·L-1NAA and30%sucrose and solidified with0.7%agar. At the same time, most callus have a lot of root hair and the rooting rate were81.7%.(7) The rooting rate were100%on1/2MS basal salts and vitamins and1%sucrose and solidified with0.7%agar. The regenerants with roots can all survive after they were transplanted to soil.(8) The regeneration period of Puccinellia tenuiflora was about16-20weeks.