A Preliminary Study On The Effect Of Genetic Transformation Of Sugar Beet BvPCNA-like Gene On The Growth Of Arabidopsis Thalian

Proliferating nuclear antigen genes are widely found in plants,but their functions in plants have been poorly studied.So far,it can only be inferred that they are related to growth and development.The research group previously obtained a gene locus of this type in the beet genome,named BvPCNA-like gene,and cloned the gene.In this study,the cloning vector carrying the exact sequence of the BvPCNA-like gene was used as the initial material to construct a set of plant expression vectors of genes for subsequent studies,and these vectors were used to transform Arabidopsis thaliana.By observing the traits of transgenic strains,the function of target genes was preliminarily analyzed,which laid the foundation for the subsequent systematic study of genes.The main research contents and results are as follows:A set of plant expression vectors for gene overexpression and expression inhibition were constructed.In this study,the overexpression vector pF-BvPCNAOE and RNAi expression inhibition vector pF-BvPCNAi of BvPCNA-like genes were designed and constructed.At the same time,due to the immature genetic transformation system of beet,the efficiency of direct use of beet RNAi vector for heterologous interference in model plants such as Arabidopsis thaliana could not be determined.In order to deduce the gene function as soon as possible,the homologous gene At PCNA1 of BvPCNA-like gene in Arabidopsis Thaliana was found and its RNAi vector pF-At PCNAi was constructed.The above three vectors were based on pFGC5941,and the upstream promoter of the inserted fragment was Ca MV35S,which was labeled as herbicide resistance gene Blp R by plant screening.Among them,the inserted fragments of RNAi vector were composed of forward target sequence,intermediate fragment and reverse target sequence,with the length of pF-BvPCNAi target sequence being 320 bp and pF-At PCNAi target sequence being 200 bp.The effects of transgenic Arabidopsis thaliana on plant growth were observed.The constructed pF-BvPCNAOE overexpression vector and pF-At PCNAi vector were infected with Arabidopsis inflorescences by Agrobacteria-mediated method for genetic transformation,and the plants resistant to phosphine oxalammonium and positive PCR results were obtained as transgenic plants.The T₂generation transgenic homozygous Arabidopsis thaliana was used as the trait analysis material.To observe the differences in traits of Arabidopsis thaliana seedlings with different genotypes.It was observed that the germination rate of overexpressed Arabidopsis thaliana was higher than that of wild type and RNAi,and the root length of overexpressed Arabidopsis thaliana seedlings was higher than that of wild type and RNAi,but the fresh weight of overexpressed Arabidopsis seedlings was lower than that of wild type and RNAi.The results showed that the increase of PCNA gene expression promoted seed germination and root growth and inhibited the growth of aboveground part..