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Expression of One Phytochrome B-Regulated NBS-LRR Gene and Preliminary Analyses of Its Functions in Rice

Author: YangHui
Tutor: MaHuiQuan；XieXianZhi
School: Shandong University of Technology
Course: Biochemistry and Molecular Biology
Keywords: Rice phytochrome NBS-LRR gene expression functional analysis
CLC: S511
Type: Master's thesis
Year: 2011
Downloads: 32
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Abstract


We compared gene profiles among wild-type, phyA and phyB mutants grown under continuous red light or in the dark. One gene, which encodes NBS-LRR family protein, is specifically up-regulated by red light perceived by phyB. We named this gene PB-LRR (phyB-regulated NBS-LRR).To access the biological functions of PB-LRR gene in rice growth, we analyzed the organ-specificity of PB-LRR expression as well as effects of exogenous hormones on PB-LRR expression. The level of PB-LRR transcripts was highest in leaf and lowest in shoot. Our results showed that light-induced expression of PB-LRR gene was attributed to phyB, not phyA. Moreover, expression of PB-LRR gene was inhibited by exogenous ABA treatment, and also regulated by environmental factors. The level of PB-LRR transcripts was not affected by PEG but probably influenced by photoperiod. The expression cassette of putative promoter region of PB-LRR and GUS (ProPB-LRR:GUS) was constructed and introduced into wild-type (WT) and phyB mutant, respectively. GUS activities were obviously detected in upper part of leaves in the WT background, but only weakly detected in the phyB mutant background. These results demonstrate that PB-LRR gene is not only regulated by phyB-mediated light signals but also by leaf development in rice.In order to analyze the PB-LRR role in rice in detail, we produced two kinds of transgenic lines:PB-LRR OE/WT (overexpression) and PB-LRR RNAi/WT (RNA interference) by Agroinfection method. Comparison of phenotypes of 14-day-old wild-type and PB-LRR transgenic seedlings revealed that the seminal roots of PB-LRR OE/WT were significantly longer than wild-type and PB-LRR RNAi/WT. No obvious difference was observed in the above-ground parts of wild type and PB-LRR transgenic lines. In addition, PB-LRR RNAi/WT showed obvious increased sensitivity to drought stress. The drought tolerance of PB-LRR OE/WT lines was similar to wild-type plants. These results suggest that PB-LRR gene is involved in regulating the rice drought tolerance.Bioinformatics analysis revealed that PB-LRR gene belonged to CC-NBS-LRR gene family. In this family, most members have been reported to be resistance gene. Thus, it was deduced that PB-LRR gene probably play a role in rice disease resitance. However, the PR-1 protein levels have no difference in wild-type and PB-LRR OE/WT leaves treated with exogenous JA and SA using western blotting analysis. These results suggest that the deduced disease resistance of PB-LRR has no relationship with expression of PR-1 proteins induced by either JA- or SA- defense pathway.Our results underlie the knowledge for further understanding the roles of PB-LRR in phyB-mediated signaling, disease resistance and drought tolerance in rice.

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