A recombinant flagellin fragment, which includes the epitopes flg22 and flgII-28, provides a useful tool to study flagellin-triggered immunity
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Plants and animals independently evolved the ability to recognize flagellin (also called FliC), the building block of the bacterial flagellum, as part of their innate immune response. While animals recognize a relatively large region of FliC, most plants recognize one or two short epitopes of FliC: flg22 and flgII-28. However, since most research in plants has focused on flg22 and flgII-28 and not the actual FliC protein, the importance of any FliC region beyond the two epitopes in plant immunity is poorly understood. Here we report cloning, overexpression, and purification of a Pseudomonas syringae FliC fragment from amino acid 1 to 143, which includes both FliC epitopes and the adjacent alpha helices. Exposing Arabidopsis thaliana leaves to FliC1–143 did not reveal any additional FliC recognition capabilities beyond flg22. However, while the kiwifruit species Actinidia arguta did not respond to either flg22 or flgII-28, treatment of A. arguta leaves with FliC1–143 triggered a significant reactive oxygen response, indicating recognition. This result suggests that in some plant species, recognition of FliC requires regions of FliC beyond the two well-known epitopes and that FliC1–143 represents a useful tool in the study of plant immunity.
KeywordsFlagellin FliC Kiwifruit Reactive oxygen species PAMP
This work was supported by NSF IOS-0746501 and IOS-1354215 to BAV. BAV was also funded in part by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, US Department of Agriculture. CRC was supported by a National Institute of Food and Agriculture Postdoctoral Research Fellowship (2015-67012-22821). NE-L was supported by BARD, the United States–Israel Binational Agricultural Research and Development Fund, Vadia-BARD Postdoctoral Fellowship Award No. FI-538-15. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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