Construction Of Plant Expression Vector And Functional Verification Of Zj2-CPgene Of Ziziphus Jujube

Jujube (Zizyphus jujuba) belongs to the plants of the genus rhamnaceae jujube, also known as the Chinese jujube,red jujube,common jujube. As early as three thousand years ago the jujube is a major economic tree species in China, the jujube has germplasm resources, almost distributes every provinces across the country, and is mainly distributed in the north of the huaihe river and qinling region. The physiological characteristics of the Jujube seedlings after the stress for drought and salt, found that the enzyme activity of jujube seedling in different tissue was higher than the enzyme activity in the control group. Real-time fluorescent quantitative results showed the relative expression of gene were higher than the control group with the increase of stress time.It shows that the2-cysteine oxidation reduction enzyme gene can improves the plant’s drought resistance and salt tolerance.The gene sequence analysis of the jujube’s2-cysteine oxidation reduction enzyme gene showed that the sequence length was834bp and encoded277amino acids, the molecular weight was30.65KD, the theoretical isoelectric point was8.95, the content of CG was45.68%, the unstability coefficient was41.56,the liposoluble index was87.29, the average of hydrophobicity was-0.12, the gene encoding protein was unsecretory and outside membrane, the homology that gene encoding protein and2-cysteine oxidation reduction enzyme gene encoding protein of the known species was82%-99%, so the gene named jujube2-cysteine peroxiredoxin (Zizyphus jujuba2-cys peroxiredoxin, Zj2-CP),the registration number in DDBJ/EMBL/GenBank was:AB812086.We put the successful plant expression vector was buided turn into the agrobact-erium cells by the freezing and thawing method, the positive bacterial strains was obtained with the kanamycin screening, we transfered the positive bacterial strains in to arabidopsis thaliana with the agrobacterium mediated, the resulted is that received15pure and transgenic arabidopsis plants after the kanamycin screening.The Laser Scanning Confocal Microscopy showed that Zj2-CP gene had different expression in the different tissue parts, roots, stems, leaf for the transgenic arabidopsis.Transgenic arabidopsis and wild type arabidopsis to drought stress and salt stress, the results of the physiological characteristics of POD, CAT, APX,MDA,showed that the enzyme activity in the wild-type arabidopsiswere was higher than the enzyme activity in the transgenic arabidopsis. By real-time fluorescent quantitative techniques showed that the wild type arabidopsis and transgenic arabidopsis after drought and salt stress,the Zj-CP in the stress group expression quantity of were higher than the control group, Zj-CP gene could improve the function of drought resistance and salt resistance for plant.Transgenic arabidopsis and wild-type arabidopsis seeds and plant were stress treatment, It found that transgenic arabidopsis’s growth higher than that of wild-type arabidopsis growth, in different salt50mmol.L-1-200mmol.L-1. Transgenic arabidopsis and wild-type arabidopsis growth were similar under the controlled conditions, transgenic arabidopsis and wild-type arabidopsis and don’t sprout, in the300mmol.L-1,400mmol.L-1, It could improve the salt resistance of transgenic arabidopsis under the condition of proper salt concentration.