Cloning And Function Analysis Of Maize Stress Tolerance Relate Gene Zm-Remorin

Continuously increasing the production of Maize (Zea maize L.) which is one of the most important food crops has become the essential step to ensure world food security. During the whole growth period of maize, it has to face to a variety of biological and abiotic stress, which will affect the plant growth and development severely, or even lead the field to fail completely. Viruses, bacteria, fungi and insects belong to biological stress, while abiotic stress includes high temperature, drought, high salt, and freezing cold injury, etc. For this affection, plant cannot escape like animals, and the only measure is to adjust its physiological and biochemical processes to adapt and resist the effects and improve their resilience defense mechanism. Maize has evolved a unique defense response mechanism which plays an important role to resist environmental stress. Therefore, it will be the important theoretical and practical significance for the entire maize production to discover and isolate the key protein genes which act on the defense response, the main contents are as follows:1、On the basis of proteomics analysis, this study focused on one of the major proteins differentially expressed genes Zm-Remorin. The results showed that full-length cDNA sequence is 1075bp, including ORF sequences of full-length 657bp, which can encode 199 amino acids,and the DNA sequence contains five exons and four introns. Zm-Remorin has the Coiled-coil module which is typical protein of remorin family, and N terminal proline content is 8.9%, so belongs to remorin la protein family group. Phylogenetic trees analysis revealed Zm-Remorin protein sequence and sorghum have a near relationship, while are far with tomato and Arabidopsis. RT-PCR analysis showed that Zm-Remorin is expressed among the various organizations, indicating that this gene may be widely involved in numerous biological processes in maize. By studying the Zm-Remorin’s expression patterns under the treatment with hormones (SA, JA and ABA), drought, high salt and SCMV virus infection by Real-time PCR technique, we found that hormones, drought and high salt can induce the expression level of Zm-Remorin, but SCMV virus infection made the expression an opposite trend in the resistance varieties, which is upregulated, and in the susceptible types showed the overall downward trend, which promise that Zm-Remorin plays an important role in biotic and abiotic stress in maize.2、We transformed a fusion expression vector pCAMBIA 1304-Zm-Remorin-GFP which is constructed by recombinant DNA technology into the model plant Arabidopsis thaliana. By observing root tip cells in transgenic Arabidopsis under the Blu-ray excitation by confocal microscopy, we found that Zm-Remorin is a membrane localization protein, indicating that Zm-Remorin acts an important role in signal transduction between the extracellular and intracellular of maize.3-. We got the transgenic plants by transform the overexpression vector pCAMBIA 1304-Zm-Remorin into Arabidopsis thaliana transgenic, and treated the offspring with different biotic and abiotic stress.The result showed that transgenic Arabidopsis will significantly improve the ability to resistant the drought, salt and anti-biological infection. Real-time PCR analysis in transgenic offspring showed that under drought and high salt conditions, and the expression level of ABA 1 and ABA2 in transgenic plants are significantly higher than the wild-type and empty vector transfected Arabidopsis, indicating Zm-Remorin may be regulate the ability of plants to drought and salt resistance through the ABA signaling pathway; Under the biological stress of Fusarium graminearum infection, the expression levels of PR1 and PR4 genes in the transgenic Arabidopsis thaliana are significantly higher than the other two genotypes, indicating Zm-Remorin may be adjust the capacity of the plant resistance to biotic stress by regulating the expression of plant defense genes PR1 and PR4.