期刊
PLANT CELL
卷 29, 期 7, 页码 1571-+出版社
AMER SOC PLANT BIOLOGISTS
DOI: 10.1105/tpc.17.00047
关键词
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资金
- National Institutes of Health [GM097587]
- National Science Foundation [IOS-1354434]
- National Research Foundation of Korea (NRF) in the Ministry of Science, ICT, and Future Planning [NRF-2015R1A2A1A01002327]
- Direct For Biological Sciences [1354434] Funding Source: National Science Foundation
- Division Of Integrative Organismal Systems [1354434] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [1549580] Funding Source: National Science Foundation
Pathogenic gram-negative bacteria cause serious diseases in animals and plants. These bacterial pathogens use the type III secretion system (T3SS) to deliver effector proteins into host cells; these effectors then localize to different subcellular compartments to attenuate immune responses by altering biological processes of the host cells. The fluorescent protein (FP)-based approach to monitor effectors secreted from bacteria into the host cells is not possible because the folded FP prevents effector delivery through the T3SS. Therefore, we optimized an improved variant of self-assembling split super-folder green fluorescent protein (sfGFP(OPT)) system to investigate the spatiotemporal dynamics of effectors delivered through bacterial T3SS into plant cells. In this system, effectors are fused to 11th beta-strand of super-folder GFP (sfGFP11), and when delivered into plant cells expressing sfGFP1-10 beta-strand (sfGFP1-10(OPT)), the two proteins reconstitute GFP fluorescence. We generated a number of Arabidopsis thaliana transgenic lines expressing sfGFP1-10(OPT) targeted to various subcellular compartments to facilitate localization of sfGFP11-tagged effectors delivered from bacteria. We demonstrate the efficacy of this system using Pseudomonas syringae effectors AvrB and AvrRps4 in Nicotiana benthamiana and transgenic Arabidopsis plants. The versatile split sfGFP(OPT) system described here will facilitate a better understanding of bacterial invasion strategies used to evade plant immune responses.
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