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Functional Analysis of A Rice Bacterial Blight Disease Resistance Gene OsNLR1

Author: DongRuiXian
Tutor: ChenJianPing
School: Zhejiang Normal University
Course: Botany
Keywords: Rice Blight Agrobacterium -mediated transformation NBS-LRR OsNLR1
CLC: S511
Type: Master's thesis
Year: 2011
Downloads: 21
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Abstract

By the Gram-negative bacteria Xanthomonas rice varieties (Xanthomonas oryzae pv. Oryzae) caused by bacterial blight (Bacterial Blight, BB) is the most serious diseases of rice one. Currently, the most effective way to control the disease is resistant gene selection and cultivation of resistant varieties. Wild rice (Oryza meyeriana L.) against Xanthomonas oryzae showed immunity, but it is no ordinary sexual hybridization between cultivated rice barriers, it is difficult for the use of their resistance genes. Y73 is meyeriana and cultivated rice susceptible \. In order to study the resistance of Agrobacterium Y73, Y73, and we have a large grain fragrant transfection efficiency were compared and analyzed. In order to facilitate the resistant materials Y73 resistance gene function in the analysis, we also need to understand the resistance of Agrobacterium Y73, Y73 transgenic establish an efficient system. In transgenic process, we co-cultured AAM suspension culture and to try to improve the optimization of temperature Y73 transgenic efficiency. Studies have shown the same transgenic conditions resistant materials Y73 transgenic efficiency (35.7%) was significantly lower than the susceptible big grain incense (71.2%), indicating that Y73 may exist some with large grain incense different resistance genes, can farmers mediated transgenic Bacillus process impact, this resistance may be associated with bacterial blight resistance. By lowering the temperature and the use of co-culture medium AAM3 suspension, we will Y73 conversion efficiency increased from 29.9% to 66.5%, so as to establish an efficient system of Y73 transgene. We found that the culture temperature of the Y73 greater impact on efficiency of gene transfer, temperature (20 ℃) ??can weaken the resistance of Agrobacterium Y73, produce higher conversion efficiency. Agrobacterium infection process in order to study the molecular mechanisms of Y73, we used qRT-PCR methods for different conversion conditions Y73 in OsMPKs, OsVIP1s and OsPR1s gene expression were studied and analyzed. In Agrobacterium-mediated gene transfer process, the activation of protein in Arabidopsis MAPK3 can make VIP1 protein phosphorylation and navigate to the plant cell nucleus, thereby regulating the PR1 gene expression, whereas VIP1 protein is also involved in the T-DNA of Agrobacterium nuclear import process. qRT-PCR studies have shown that two OsVIP1s and almost all OsPR1s 20 ℃ Agrobacterium gene expression after infection was higher than 25 ℃, while OsMPKs no significant change. We speculate that under low temperature conditions and VirE2 OsVIP1 induced protein interactions, thereby facilitating complex Agrobacterium T-DNA transfer to plant cells, leading to a higher conversion efficiency. The study also showed that the presence in rice and Arabidopsis defense signal path similar MAPK/VIP1 participate Agrobacterium mediated plant transgenic process. In Y73 in the signaling pathway may be affected by the regulation of temperature, thereby affecting Y73 Agrobacterium-mediated gene transfer efficiency. Process in plant disease resistance genes play an important role in disease resistance genes encode a large part of the current product has NBS-LRR domain, the structure and the interactions between proteins and disease is closely related to signal transduction. Our previous gene expression microarray analysis showed high resistance to bacterial blight of rice new varieties Y73 blight strains inoculated P10 (PXO124) after an NBS-LRR genes significantly up-regulated expression, we used RT- Y73 was cloned by PCR from CDS of the gene fragment (2631 bp), and named this gene OsNLRl (Rice NBS-LRR Resistance gene 1). Sequence analysis showed that the amplified fragment was consistent with the sequence of cultivated rice, we hypothesized that the gene may be affected by O.meyeriana impact resistance genes in rice disease resistance and participate in the process. In order to analyze its function, we use pCAMBIA13000 binary vectors were constructed OsNLR1 gene overexpression and interference vectors, and using Agrobacterium-mediated gene transfer method to obtain transgenic rice plants. Inoculation experiments showed OsNLR1 inhibiting gene expression, can weaken the resistance of the resistance material Y73; OsNLR1 gene expression and increased levels of susceptible material is able to enhance the resistance to disease resistance. These results indicate that OsNLR1 rice bacterial blight resistance gene and directly related to In addition, OsNLR1 subgenotype cellular localization results also show, OsNLR1 coding products mainly located in the cell membrane and the nucleus of rice, indicating that the gene may be related to signal transduction and transcription of downstream genes are closely related.

Functional Analysis of A Rice Bacterial Blight Disease Resistance Gene OsNLR1

Abstract

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