| ABSTRACT:SO2is the main pollutant of the atmosphere in China which is harmful to crops such as corn, also is an important constraint to maize’s and other crops’yield and quality. At present, the research of the impact of SO2to the plant, mainly from injury symptoms, mechanism, physiology, biochemistry, growth and development, as well as the compounded impact of SO2and other pollutants to the plants. In this article, the research we did was cloning and function Analysis of ZmpsbA gene from Maize in response to SO2Stress from nucleic acid.First, using the technology of DDRT-PCR, study the differentially expressed genes in the inbred lines of ordinary maize Q319under SO2stress, obtained valuable differentially expressed fragments. Second, verification of fragments of differentially expressed genes expression under different conditions, analysis of its relationship with the SO2stress. At last, using the technology of Homology cloning,get the full-length gene.verification and analysis the function of the full-length gene and the corresponding relationship with the SO2stress.The results of our research are as follows:1. Fourteen differentially expressed gene fragments were obtained, of which Zp, Dl, D3, D5, D6and D11have been searched homology sequence, the other eight are not.In the three cDNA fragments of Enhanced expression,the protein similarity of Zp compared with psbA which encoding photosystem Ⅱ (PS Ⅱ) reaction center of the32kDa protein is up to99%; D1compared with ABP which encoding amino acid binding protein is up to82%; D5a compared with DOF which encoding Zinc finger transcription factor protein)zsimilar is up to91%.2. From the expression of Zp under different conditions of SO2and NaHSO3stress, finding that the mRNA expression levels of the gene in the inbred line Q319of maize are significantly increased.3. By the homology cloning, obtained the full-length gene ZmpsbA which contained the fragment of Zp.ZmpsbA has1062nucleotides and encoding352amino acids.4. By the work of ZmpsbA’s transgenic, at last we get six Overexpression transgenic lines and one silence expression transgenic line of tobacco.In the process of ZmpsbA silence expression transgenic line’s cultivation,we found that it is more sensitive to high temperature.In addition,although it can be flowering normaly,but the fruit’s fertility isn’t normal.5. By the analysis of the ZmpsbA overexpression transgenic lines which under theSO2stress; we found that transgenic lines are extremely sensitive to SO2. AT the phenotypic, the transgenic lines’degree of injury is more than wild-type control.AT the molecular, before and after treatment, the mRNA volume of ZmpsbA expression in the transgenic lines or the wild type controls, have been significant changed.especially after the treatment of24h, the mRNA volume of ZmpsbA expression in the wild-type control in a downward trend, while in the transgenic lines into a clear upward trend.6. In addition, we are also analysis many tobacco genes’ expression which sensitive to stress in transgenic lines and wild-type controls under SO2stress.Found that some of them involved in the response process.Through the research of ZmpsbA, we believe that genes involved in the SO2stress response process of mays. The level of plant Injury with the level of gene expression and duration are closely related. The gene is also common with other genes involved in the stress response of SO2stress, but the regulation of the ZmpsbA gene will be play an important role. The gene may be sensitive to the strength and with the relationship to fertility. |
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