OsPRK Enhances Drought Tolerance and Blast Resistance in Rice (Oryza sativa) via Metabolic Reprogramming
HE Wei1,2,3, ZHOU Ping4, WEI Yi-Dong1,2,3, CUI Li-Li1,2,3, CAI Qiu-Hua1,2,3, XIE Hong-Guang1,2,3, XIE Hua-An1,2,3, ZHANG Jian-Fu1,2,3,*
1 Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350019, China; 2 State Key Laboratory for Ecological Control of Crop Pests between Fujian and Taiwan, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China; 3 National Engineering Laboratory of Rice/South China Research Base of State Key Laboratory of Hybrid Rice/Incubating Base of State Key Laboratory of Crop Germplasm Innovation and Molecular Breeding between Fujian Province and Ministry of Science and Technology/Fuzhou Branch of National Rice Improvement Center/South China Key Laboratory of Hybrid Rice Germplasm innovation and Molecular Breeding of Ministry of Agriculture and Rural Areas/Fujian Engineering Laboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding, Fuzhou 350003, China; 4 Fruit Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China
Abstract：Phosphoribulokinase (PRK) is a key enzyme in the Calvin cycle of photosynthetic reactions and plays a central role in carbon assimilation. Recent studies suggest that PRK may also participate in plant responses to abiotic and biotic stresses, but its molecular mechanisms under stress conditions remain unclear. This study investigated the regulatory mechanism of OsPRK in response to drought and rice blast fungus (Magnaporthe oryzae) stress by generating OsPRK-overexpressing transgenic rice lines and combining metabolomic analysis with phenotypic characterization. Using Agrobacterium-mediated genetic transformation and qPCR, we successfully obtained T2 generation homozygous transgenic rice plants (Oryza sativa ssp. japonica cv. Nipponbare) carrying a single-copy transgene and exhibiting significantly higher OsPRK expression than wild-type (WT) plants. High-performance liquid chromatography-tandem mass spectrometry approach were employed to analyze metabolites in three-leaf-stage seedlings from transgenic and WT plants, identifying a total of 2 036 metabolites. Orthogonal partial least squares-discriminant analysis screened 136 significantly differentially accumulated metabolites (DAMs), including 93 that were upregulated and 43 that were downregulated. These DAMs were primarily enriched in metabolic pathways potentially related to stress response, such as isoflavonoid biosynthesis, flavonoid biosynthesis, and phenylpropanoid biosynthesis. Phenotypic analysis under stress conditions showed that OsPRK-overexpressing plants exhibited delayed leaf wilting under drought stress and had approximately 17% higher survival rate than WT plants after rehydration. After inoculation with Magnaporthe oryzae, transgenic plants developed a significantly fewer lesions, indicating enhanced resistance. Furthermore, key photosynthetic parameters at the mature stage, including net photosynthetic rate and stomatal conductance, were not significantly affected, demonstrating that OsPRK overexpression enhances stress tolerance without compromising photosynthetic efficiency. Interestingly, it was observed that T0 generation osprk mutant plants exhibited significantly lower chlorophyll content during the seedling hardening stage compared with WT and overexpressing lines, and eventually died. This study reveals a novel mechanism by which OsPRK regulates stress resistance in rice through metabolic networks remodeling, providing a theoretical foundation for molecular breeding of stress-tolerant crops.
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