Abstract
This chapter describes methods for targeted knockouts using marker exchange mutagenesis and complementation of the Gram-negative bacteria Xanthomonas oryzae pv. oryzae. We have used these methods to demonstrate that type I secretion and modification systems are involved in avrXa21 activity of X. oryzae pv. oryzae.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Girardin, S., Sansonetti, P. J., and Philpott, D. J. (2002) Intracellular vs extracellularrecognition of pathogens—common concepts in mammals and flies. Trends Microbiol. 10, 193–199.
Janeway, C. A., Jr. and Medzhitov, R. (1997) Innate immune recognition. Annu. Rev. Immunol. 20, 197–216.
van’t Slot, K. A. E. and Knogge, W. (2002) A dual role for microbial pathogen-derived effector proteins in plant disease and resistance. Crit. Rev. Plant. Sci. 21, 229–271.
Medzhitov, R. (2001) Toll-like receptors and innate immunity. Nat.Rev. Immunol. 1, 135–145.
Hugot, J., Chamaillard, M., Zouali, et al. (2001) NOD2 leucine-rich repeat variants with susceptibility to Crohn’s disease. Nature 411, 599–603.
Werling, D. and Jungi, T. (2003) TOLL-like receptors linking innate and adaptive immune response. Vet. Immunol. Immunopathol. 91, 1–12.
Inohara, N. and Nunez, G. (2003) Nods: intracellular proteins involved in inflammation and apoptosis. Nat. Rev. Immunol. 3, 371–382.
Werling, D. and Jungi, T. W. (2003) TOLL-like receptors linking innate and adaptive immune response. Vet. Immunol. Immunopathol. 91, 1–12.
Barton, G. M. and Medzhitov, R. (2003) Toll-like receptor signalling pathway. Science 300, 1524–1525.
Hayashi, F., Smith, K., Ozinsky, A., et al. (2001) The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410, 1099–1103.
Hoshino (1999) Cutting edge: Toll-like receptor 4-deficient mice are hyporesponsive to LPS. J. Immunol. 162, 3749–3752.
Girardin, S., Boneca, I., Carneiro, L., et al. (2003) Nod1 detects a unique muropeptide from gram-negative bacterial peptidoglycan. Science 300, 1584–1587.
Jones, D., Thomas, C., Hammond-Kosack, K., Balint-Kurti, P., and Jones, J. (1994) Isolation of the tomato cf-9 gene for resistance to Cladosporium fulvum by transposon tagging. Science 266, 789–793.
Wang, G., Ruan, D., Song, W., et al. (1998) Xa21D encodes a receptor-like molecule with a leucine-rich repeat domain that determines race-specific recognition and is subject to adaptive evolution. Plant Cell 10, 765–779.
Gomez-Gomez, L. and Boller, T. (2002) Flagellin perception: a paradigm for innate immunity. Trends Plant Sci. 7, 251–256.
Shiu, S.-H. and Bleecker, A. B. (2001) Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proc. Natl. Acad. Sci. USA 98, 10,763–10,768.
Gomez-Gomez, L. and Boller, T. (2000) FLS2: an LRR-receptor like kinase protein involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Mol. Cell 5, 1003–1011.
Matsubayashi, Y., Ogawa, M., Morita, A., and Sakagami, Y. (2002) An LRR receptor kinase involved in perception of a peptide plant hormone, phytosulfokine. Science 296, 1470–1472.
Yin, Y., Wu, D., and Chory, J. (2002) Plant receptor kinases: Systemin receptor identified. Proc. Natl. Acad. Sci. USA 99, 9090–9092.
Cock, J. M., Vanoosthuyse, V., and Gaude, T. (2002) Receptor kinase signalling in plants and animals: distinct molecular systems with mechanistic similarities. Curr. Opin. Cell Biol. 14, 230–236.
Che, F.-S., Nakajima, Y., Tanaka, N., et al. (2000) Flagellin from an incompatible strain of Pseudomonas avenae induces a resistance response in cultured rice cells. J. Biol. Chem. 275, 32,347–32,356.
Shen, Y., Chern, M., Silva, F. G., and Ronald, P. (2001) Isolation of a Xanthomonas oryzae pv. oryzae flagellar operon region and molecular characterization of flhF. Mol. Plant Microbe Interact. 14, 204–213.
Shen, Y., Sharma, P., da Silva, F. G., and Ronald, P. (2002) The Xanthomonas oryzae pv. lozengeoryzae raxP and raxQ genes encode an ATP sulphurylase and adenosine-5′-phosphosulphate kinase that are required for AvrXa21 avirulence activity. Mol. Microbiol. 44, 37–48.
Burdman, S., Shen, Y., Lee, S.-W., Xue, Q., and Ronald, P. (2004) RaxH/RaxR: a two-component regulatory system in xanthomonas oryzae pv. oryzae required for AvrXa21 activity. MPMI 17, 602–612.
Goes da Silva, F., Shen, Y., Dardick, C., et al. (2004) Components of a type I secretion system and a sulfotransferase-like protein are required for the Xa21 receptor kinase mediated defense response. Mol. Plant Microbe Interact. 17, 593–601.
Labes, M., Puhler, A., and Simon, R. (1990) A new family of RSF1010-derived expression and lac-fusion broad-host-range vectors for gram-negative bacteria. Gene 30, 37–46.
DeFeyter, R., Kado, C., and Gabriel, D. (1990) Samll, stable shuttle vector for use in Xanthomonas. Gene 88, 65–72.
Sambrook, J., Fritsch, E., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc.
About this protocol
Cite this protocol
Lee, SW., Ronald, P.C. (2007). Marker-Exchange Mutagenesis and Complementation Strategies for the Gram-Negative Bacteria Xanthomonas oryzae pv. oryzae . In: Ronald, P.C. (eds) Plant-Pathogen Interactions. Methods in Molecular Biology, vol 354. Humana Press. https://doi.org/10.1385/1-59259-966-4:11
Download citation
DOI: https://doi.org/10.1385/1-59259-966-4:11
Publisher Name: Humana Press
Print ISBN: 978-1-58829-448-7
Online ISBN: 978-1-59259-966-0
eBook Packages: Springer Protocols