Abstract
Type-III (T3) effectors PthXo1 and AvrXa10 of Xanthomonas oryzae pv. oryzae are translocated into rice cells to induce virulence and avirulence on susceptible- and resistant-rice varieties Nipponbare and IRBB10, respectively. The translocation needs the bacterial T3 translocator Hpa1 and rice Oryza sativa plasma membrane protein OsPIP1;3. Here, we employed the β-lactamase (BlaM) reporter system to observe PthXo1 and AvrXa10 translocation. The system was established to monitor effectors of animal-pathogenic bacteria by quantifying the BlaM hydrolysis product [P] and fluorescence resonance energy transfer (FRET) of the substrate. The feasibility of the BlaM reporter in rice protoplasts was evaluated by three criteria. The first criterion indicated differences between both [P] and FRET levels among wild types and OsPIP1;3-overexpressing and OsPIP1;3-silenced lines of both Nipponbare and IRBB10. The second criterion indicated differences between [P] and FRET levels in the presence and absence of Hpa1. The last criterion elucidated the coincidence of PthXo1 translocation with induced expression of the PthXo1 target gene in protoplasts of Nipponbare and the coincidence of AvrXa10 translocation with induced expression of the AvrXa10 target gene in protoplasts of IRBB10. These results provide an experimental avenue for real-time monitoring of bacterial T3 effector translocation into plant cells with a pathological consequence.
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Abbreviations
- BlaM:
-
β-lactamase
- PIP:
-
plasma membrane intrinsic protein
- PM:
-
plasma membrane
- TAL:
-
translocation activator-like
- T3:
-
type III
- Xoo :
-
Xanthomonas oryzae pv. oryzae
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Acknowledgements
We thank Professor Bing Yang (Department of Genetics, Development, and Cell Biology, Iowa State University) for the gift of the pthXo1 vector and our colleague Professor Ling Jiang (Agronomy College, Nan**g Agricultural University) for assistance in rice transformation. We thank financial support from the China National Key Research and Development Plan (Grant Number 2017YFD0200901), Natural Science Foundation of China (Grant Number 31772247) and Talent Recruitment Funding of Shandong Agricultural University (Grant Number 20171226).
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Communicated by Ashis Kumar Nandi.
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Bian, H., Zhang, L., Chen, L. et al. Real-time monitoring of translocation of selected type-III effectors from Xanthomonas oryzae pv. oryzae into rice cells. J Biosci 44, 82 (2019). https://doi.org/10.1007/s12038-019-9916-0
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DOI: https://doi.org/10.1007/s12038-019-9916-0