Studies On Transformation Systems Of Adiantum Reniforme Var.Sinense And Platycerium Wallichii

In this experiment, Adiantum reniforme var. sinense was used as material to study the effect of different hormones on A. reniforme var. sinense prothallium proliferation, and the normal differentiated sporophyte rhizome was selected as material to investigate the effect of different growth regulators on the GGB pathway under laboratory conditions, including the GGB induction, proliferation and differentiation; Platycerium wallichii spore leaf was chosed as material to investigate the effect of different hormones on the GGB induction, GGB proliferation and differentiation, the rooting induction of adventitious bud. Finally, we tried to use A. reniforme var. sinense prothallium, Platycerium wallichii GGB as receptor materials to grope the genetic transformation system of two kinds of ferns by Agrobacterium tumefaciens mediated method. The main results were as follows:GGB was induced by A. reniforme var. sinense sporophyte rhizom, successfully establishing the A. reniforme var. sinense regeneration pathway through GGB. The results showed that the optimum medium inducing A. reniforme var. sinense GGB was MS+BA0.5mg/L+2,4-D1.0mg/L, and the induction rate reached55.56%, while reproduced medium for A. reniforme var. sinense GGB proliferation was MS+6-BA1.0mg/L+NAA0.1mg/L, medium of differentiation for Adiantum GGB was MS+ZTO. lmg/L, and the differentiation rate reached38.29%; meanwhile, proliferation medium of A. reniforme var. sinense reniforme prothallium was optimized on the previous research basis, the optimum medium obtained from the experiment was MS+6-BA1.0mg/L+IBAO.1mg/L.Compared with the A. reniforme var. sinense, Platycerium wallichii GGB was induced more easily, and the proliferation and differentiation rate of GGB were high. The results showed that the induction rate could reach to100%in MS+BA1.0mg/L+NAA1.0mg/L and MS+BA2.0mg/L+NAA0.5mg/L medium, neither the placed way nor the blade surface scratched or no of experimental material have effect on the induction of GGB, medium for staghorn fern GGB proliferation was MS+KT1.0mg/L+NAA0.1mg/L, the staghorn fern GGB differentiation rate could reach to100%on the MS medium, in addition, medium for rooting of Platycerium sporophytes was MS+NAA0.5mg/L.Finally, genetic transformation system of Adiantum fern was established preliminarily:Using A. reniforme var. sinense prothallus as material, preincubation and co-culture medium were MS+BA1.0mg/L+IBAO.1mg/L; the selecting medium was MS+BA1.0mg/L+IBA0.1mg/L+Kan100mg/L+Cef400mg/L; preincubation time for2days; OD600=0.4, infection time for10mins; after rinsing1-2times by liquid MS medium containing250mg/L Cef, the explants were inoculated on the solid MS regeneration medium containing250mg/L Cef, postponed screening15d; Then the explants were inoculated and cultured on the selection medium.