The Influence Of CTK To Maize Phyllotaxis Of Tissue Culture And The Construction Of Zmrr3 Gene RNAi Vector

The Mazie is the grasses,its phyllotaxis is living the way is the alternate growth. This laboratory has obtained the phyllotaxis opposite mutant through the mutagenesis, From the Mazie of decussate phyllotaxis mutant's research we discovered that its endogene CTK the level increasing is the key trait of decussate phyllotaxis.We add different concentrations of 6 - BA divide the Luyuan 92 embryo to research on relationship between the phyllotaxis of Maize and extrior cytokinin 6 - BA,.In order to verifying the relation between cytokinins and phyllotaxis of Maize,we construct the cytokinin Zmrr3 gene expression vector of RNAi.The results are as follows:1,Take the Luyuan92 of maize as the material,add different concentrations of extrior cytokinin 6 - BA in the medium callus,found that callus induction rate was in declining trend when increased the comcentration of exterior cytokinin 6-BA.2,Different concentration of 6 - BA the callus induced by the same culture medium, which evidently impact the sprouting rate.with 6 - BA to increase the concentration of the sprouting rate of an upward trend,and when 6 - BA concentration Less than 1.5 mg / L at the obvious differences,but when 6 - BA of more than 1.5 mg / L,which was not obvious differences,it moves toward stability.3,Influenced by different,the phyllotary of Maize which induced by different regeneration culture medium the seedlings has changed.with 6 - BA concentration of the increasing,the frequence of alternative phyllotary induced by different callus medium increases gradually,and the frequence of decussate phyllotary induced by different callus medium increases at frist,then lowers.It is full of alternative phyllotary and the sprouting rate is higher when adds 6 - BA concentration of 0.2 mg/ L in callus medium and 6 - BA concentration of 0.5 mg / L in regeneration culture medium.Which has set up a transformation for the regeneration system.4,Get the gene Zmrr3 the fifth exon-specific segments to the sequencing from Luyuan 92 of maize genome,construct the Expression Vector with RNAi expression pCAMBIA303 + RNAi +2 F,which contains Hygromycin,kanamycin kanamycin and GUS slective gene.Which can be used to study the relation between cytokinins and phyllotaxis of Maizet in molecular mechanism.In summary,this findings indicated that through inducing the tissue culture,the extraneous 6-BA may induce the transformation of Maize phyllotaxis and construct the RNAi expression Vector,The results is of great significance for revealing the phullotary of Maize in molecular mechanism...