WRKY transcription factors are crucial regulatory components of plant responses to pathogen infection. In the present study, we report isolation and functional characterization of the pathogen-responsive rice WRKY30 gene, whose transcripts accumulate rapidly in response to salicylic acid (SA) and jasmonic acid (JA) treatment. Overexpression of WRKY30 in rice enhanced resistance to rice sheath blight fungus Rhizoctonia solani and blast fungus Magnaporthe grisea. The enhanced resistance in the transgenic lines overexpressing WRKY30 was associated with activated expression of JA synthesis-related genes LOX, AOS2 and pathogenesis-related (PR)3 and PR10, and increased endogenous JA accumulation under the challenge of fungal pathogens. WRKY30 was nuclear-localized and had transcriptional activation ability in yeast cells, supporting that it functions as a transcription factor. Together, our findings indicate that JA plays a crucial role in the WRKY30-mediated defense responses to fungal pathogens, and that the rice WRKY30 seems promising as an important candidate gene to improve disease resistance in rice.
WRKY transcription factors have been implicated in the regulation of transcriptional reprogramming associated with various plant processes but most notably with plant defense responses to pathogens. Here we demonstrate that expression of rice WRKY4 gene (OsWRKY4) was rapidly and strongly induced upon infection of Rhizoctonia solani, the causing agent of rice sheath blight, and exogenous jasmonic acid (JA) and ethylene (ET). OsWRKY4 is localized to the nucleus of plant cells and possesses transcriptional activation ability. Modulation of OsWRKY4 transcript levels by constitutive overexpression increases resistance to the necrotrophic sheath blight fungus, concomitant with elevated expression of JA- and ET-responsive pathogenesis-related (PR) genes such as PR1a, PR1b, PR5 and PR10/PBZ1. Suppression by RNA interference (RNAi), on the other hand, compromises resistance to the fungal pathogen. Yeast one-hybrid assay and transient expression in tobacco cells reveal that OsWRKY4 specifically binds to the promoter regions of PR1b and PR5 which contain W-box (TTGAC[C/T]), or W-box like (TGAC[C/T]) cis-elements. In conclusion, we propose that OsWRKY4 functions as an important positive regulator that is implicated in the defense responses to rice sheath blight via JA/ET-dependent signal pathway.
BackgroundPlant WRKY transcription factors play pivotal roles in diverse biological processes but most notably in plant defense response to pathogens. Sheath blight represents one of the predominant diseases in rice. However, our knowledge about the functions of WRKY proteins in rice defense against sheath blight is rather limited.ResultsHere we demonstrate that the expression of Oryza sativa WRKY80 gene (OsWRKY80) is rapidly and strongly induced upon infection of Rhizoctonia solani, the causal agent of rice sheath blight disease. OsWRKY80 expression is also induced by exogenous jasmonic acid (JA) and ethylene (ET), but not by salicylic acid (SA). OsWRKY80-GFP is localized in the nuclei of onion epidermal cells in a transient expression assay. Consistently, OsWRKY80 exhibits transcriptional activation activity in a GAL4 assay in yeast cells. Overexpression of OsWRKY80 in rice plants significantly enhanced disease resistance to R. solani, concomitant with elevated expression of OsWRKY4, another positive regulator in rice defense against R. solani. Suppression of OsWRKY80 by RNA interference (RNAi), on the other hand, compromised disease resistance to R. solani. Results of yeast one-hybrid assay and transient expression assay in tobacco cells have revealed that OsWRKY80 specifically binds to the promoter regions of OsWRKY4, which contain W-box (TTGAC[C/T]) or W-box like (TGAC[C/T]) cis-elements.ConclusionsWe propose that OsWRKY80 functions upstream of OsWRKY4 as an important positive regulatory circuit that is implicated in rice defense response to sheath blight pathogen R. solani.Electronic supplementary materialThe online version of this article (doi:10.1186/s12284-016-0137-y) contains supplementary material, which is available to authorized users.
Aims: Isolation and characterization of nicotine‐degrading bacteria with advantages suitable for the treatment of nicotine‐contaminated water and soil and detection of their metabolites. Methods and Results: A novel nicotine‐degrading bacterial strain was isolated from tobacco field soil. Based on morphological and physiochemical properties and sequence of 16S rDNA, the isolate was identified as Pseudomonas sp., designated as CS3. The optimal culture conditions of strain CS3 for nicotine degradation were 30°C and pH 7·0. However, the strain showed broad pH adaptability with high nicotine‐degrading activity between pH 6·0 and 10·0. Strain CS3 could decompose nicotine nearly completely within 24 h in liquid culture (1000 mg L−1 nicotine) or within 72 h in soil (1000–2500 mg kg−1 nicotine) and could endure up to 4000 mg L−1 nicotine in liquid media and 5000 mg kg−1 nicotine in soil. Degradation tests in flask revealed that the strain had excellent stability and high degradation activity during the repetitive degradation processes. Additionally, three intermediates, 3‐(3,4‐dihydro‐2H‐pyrrol‐5‐yl) pyridine, 1‐methyl‐5‐(3‐pyridyl) pyrrolidine‐2‐ol and cotinine, were identified by GC/MS and NMR analyses. Conclusions: The isolate CS3 showed outstanding nicotine‐degrading characteristics such as high degradation efficiency, strong substrate endurance, broad pH adaptability, and stability and persistence in repetitive degradation processes and may serve as an excellent candidate for applications in the bioaugmentation process to treat nicotine‐contaminated water and soil. Also, detection of nicotine metabolites suggests that strain CS3 might decompose nicotine via a unique nicotine‐degradation pathway. Significance and Impact of the Study: The advantage of applying the isolated strain lies in broad pH adaptability and stability and persistence in repetitive use, the properties previously less focused in other nicotine‐degrading micro‐organisms. The strain might decompose nicotine via a nicotine‐degradation pathway different from those of other nicotine‐utilizing Pseudomonas bacteria reported earlier, another highlight in this study.
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