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Identification and Characterization of Novel Candidate Effector Proteins from Magnaporthe oryzae
Di Liu1, Zhiqin Lun1, Ning Liu1
1MOA Key Laboratory of Pest Monitoring and Green Management, China Agricultural University, Beijing 100193, China.
Abstract:
The fungal pathogen Magnaporthe oryzae secretes a large number of effector proteins to facilitate infection, most of which are not functionally characterized. We selected potential candidate effector genes from the genome of M. oryzae, field isolate P131, and cloned 69 putative effector genes for functional screening. Utilizing a rice protoplast transient expression system, we identified that four candidate effector genes, GAS1, BAS2, MoCEP1 and MoCEP2 induced cell death in rice. In particular, MoCEP2 also induced cell death in Nicotiana benthamiana leaves through Agrobacteria-mediated transient gene expression. We further identified that six candidate effector genes, MoCEP3 to MoCEP8, suppress flg22-induced ROS burst in N. benthamiana leaves upon transient expression. These effector genes were highly expressed at a different stage after M. oryzae infection. We successfully knocked out five genes in M. oryzae, MoCEP1, MoCEP2, MoCEP3, MoCEP5 and MoCEP7. The virulence tests suggested that the deletion mutants of MoCEP2, MoCEP3 and MoCEP5 showed reduced virulence on rice and barley plants. Therefore, those genes play an important role in pathogenicity.
Insights
Magnaporthe oryzae fungal effectors MoCEP2 and MoCEP5 are crucial for pathogenicity. Researchers identified and characterized these effectors, finding they induce cell death and suppress plant immune responses, leading to reduced virulence in knockout mutants.
Area of Science:
- Plant Pathology
- Molecular Biology
- Fungal Genetics
Background:
- The fungal pathogen Magnaporthe oryzae employs numerous effector proteins to infect host plants, but their functions remain largely unknown.
- Understanding these effectors is critical for developing strategies to control rice blast disease.
Purpose of the Study:
- To identify and functionally characterize novel effector genes from Magnaporthe oryzae.
- To investigate the role of selected effectors in plant cell death induction, immune suppression, and fungal virulence.
Main Methods:
- Genome-wide screening and cloning of 69 putative effector genes from M. oryzae field isolate P131.
- Transient expression assays in rice protoplasts and Nicotiana benthamiana for cell death induction and ROS burst suppression.
- Gene knockout in M. oryzae and virulence assays on rice and barley.
Main Results:
- Four effectors (GAS1, BAS2, MoCEP1, MoCEP2) induced rice cell death; MoCEP2 also induced cell death in N. benthamiana.
- Six effectors (MoCEP3-MoCEP8) suppressed flg22-induced ROS burst in N. benthamiana.
- Deletion mutants of MoCEP2, MoCEP3, and MoCEP5 exhibited reduced virulence on rice and barley.
Conclusions:
- Several novel Magnaporthe oryzae effector proteins play significant roles in pathogenicity.
- MoCEP2, MoCEP3, and MoCEP5 are essential for full virulence on host plants, highlighting their potential as targets for disease control.
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