Research On Transformation Of RNAi Expression Vector PJIM19PHYB About A.Edgeworthii Benth Into Arabidopsis And Soybean

Phytochrome B(PHYB) is a important member of the phytochrome gene families.It is involved in the regulation of physilolgical processes of plants,include shade avoidance response and photoperiodic response.The purpose of the research is transfer RNAi expression vector PJIM19PHYB about A.edgeworthii Benth into arabidopsis and soybean by agrobacerium mediated method.Observe and analyze the effect of the growth process on transgenic arabidopsis. And lay the foundation for the identification function in soybeans about the RNAi expression vector.At the same time optimize the soybean genetic transformation system,lay the foundation for soybean molecular breeding.In this experiment we analysis the funtion by transferring RNAi expression vector into arabidopsis.Optimize the soybean cotyledons node and embryonic tip regeneration system and genetic transformation system of agrobacterium-mediated soybean cotuledonary node and embryo tip.Transfering RNAi expression vector into arabidopsis and soybean preliminarily.The main research results are as follows:1.The transformation of RNAi expression vector into arabidopsisTransfor the arabidopsis by agrobacterium-mediated,after kanamycin resistance select,we get the posistive plants.The results prove that expression vector has been transferred to arabidopsis by PCR test.We fund the apparent elongation of arabidopsis hypocotyls and the flowering period in advance and reduced sensitivity of the photoperiod and branch increased and enhance shade tolerance under the photoperiod and light intensity condition.2The transformation of RNAi expression vector on the soybean cotyledon nodeSet the plant hormone concentration and ratios,optimize the soybean cotyledon node germination medium,buds induction medium,rooting medium.Screen the higher regeneration frequency of soybeans as the transformation of the material,include Gongxuanlhao,Daisidadou2,Sidou209,optimiz the agrobacterium mediated genetic transformation system of soybean cotyledonary node,determine the concentration of kanamycin is100mg/1,infection time is20min,total culture time is3days. We have got the positive plants through the transformation of soybean cotyledonary node,include Gongxuanlhao and Sidou209each have a plant.The conversion rate is0.56%and0.64%.3The transformation of RNAi expression vector on the soybean embryonic tips.Set the plant hormone concentration and ratios,optimize the soybean cotyledon node germination medium,buds induction medium,rooting medium.And optimize the genetic transformation system for soybean embryonic tips.The results show that the infection time is20min and the concentration of AgNO3is2mg/l.Use embryos for explants,include Gongxuanlhao and Daisidadou2,we have got the positive plants through the transforation of soybean embryonic tips.The conversion rate is1.32%and0.65%.