The Analysis Of DRE, W And S Elements In The Transgenic Rice

Abstract: Plants in the long process of evolution has gradually formed the inducible defense reaction mechanism,which makes the defense response for the cost of materials and energy consumption limited to a specific time or space in order to achieve effective utilization of material and energy.Defense response genes response to adversity stress is mainly controlled by the identification and integration of the cis-acting elements and their corresponding trans-acting factors (transcription factors) in the defense response gene.So, the separation and analysis of the cis-acting element of the response to specific adversity is not only conducive to further separation of the corresponding transcription factor inducible defense response to clarify the signaling transduction pathway, is also beneficial to use these components to build inducible synthetic promoter for the genetic improvement of plant resistance Reverse genetic engineering. This paper is a more systematic study,for the response to different stress role of the isolated and identified DRE , W and s-box components on predecessors from dicotyledonous plants in the rice. The main findings are as follows:1. In an ordinary endonuclease connection technology and gateway cloning techniques ,it builds a DRE, W and s-boxes, and other participants cis-acting elements inserted into the CaMV35S core promoter (+ 8bp—-46bp) upstream and controls the plant expression vector of GUS reporter gene expression, and tests the carrier by sequencing.2. Using Agrobacterium-mediated transformation methods to turn the carrier into japonica Nipponbare,it wins the vector corresponding transgenic T0 generation plants and makes T0 generation plants in the culture under-glass and makes them selfing,homozygous and hygromycin to be immersed for selecting,respectively requiring its inadequate T1 and T2 generation strains.3. Using the seeds and plants or its derivative resistance callus of these transgenic strains, at low temperature, high temperature and salinity, rice leaf roller feeding and rice blast vaccination and so on and together with the analysis methods of GUS histochemical stain,the author analyzes the response from the DRE, W and s-box components in transgenic rice on participants of different stress and analyzes the possible relationship between the response from different components to the stress and several internal endogeny hormone signals. The experiment found that W-box can be mediated the response from rice to fungus Magnaporthe grisea and, participated in a rice feeding response, and possibly with JA and SA signal pathway relationships. Gus expression driven by DRE box to the answer of the various abiotic stresses is different,,and it may be related to the upper reaches of the relationship between hormonal signals. S-box could significantly be mediated the response from rice blast and rice leaf roller ,and the level on a variety of abiotic stress response is different, while it is the external expression of a certain hormone.4. Selecting on pathogens bacteria or evoked inducer processing efficient answering components, and selecting the appropriate number of copies and appropriate combination to build the expression vector of inducible promoter containing more than one combination of CIS-acting elements (2×DRE + 2×GCC + 2×JERE + 2×W),and by Agrobacterium-mediated transformation method to obtain a transgenic plant T0 generation, identify confrontational plant by PCR.Preliminary findings indicate that between unifacial leaf rice and dicotyledonous plants, CIS-acting elements and trans-act factor between recognition and interaction activating plant defense responses exist a certain degree of conservative, W and s-box in the rice blast and rice leaf roller, and other biological adversity stress have strong response, at the same time they have different levels of response at low temperature, high salt and so abiotic stress,and this implied the existence of complex signaling pathways of crosstalk in its upper reaches ,and might have something to do with salicylic acid, Abscisic acid and other hormones involved. W and s-box, and other cis-acting elements can not rely on its flanking in different genetic backgrounds alone,and this indicates that this class of cis-acting original synthesis in inducible promoter of the construction is of great value.