Plant stomata are an essential route for bacterial pathogens to entry inside host tissue and cause diseases. As an important defense mechanism, plant stomata can actively restrict bacterial invasion by dynamically regulating the opening, closing, and reopening of stomatal guard cells. Therefore, accurately measuring the stomatal aperture size during the bacterial pathogenesis is an important approach to study the stomata related immunity. Several methods have been developed for stomatal aperture measurement. Here, we described a detailed protocol of using clear nail polish to make Arabidopsis epidermal impressions for investigate the change of stomatal aperture size in plant immune response. The application of this approach can instantly fix the status of stomatal guard cells, and provides clear, stable, and almost permanent slides of epidermal impressions for measurement of stomatal aperture size.
Nail polish Arabidopsis thalianaEpidermal impressions Stomatal guard cell
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This work was supported by the National Science Foundation.
Andrews JH, Harris RF (2000) The ecology and biogeography of microorganisms on plant surfaces. Annu Rev Phytopathol 38(1):145–180CrossRefPubMedGoogle Scholar
Beattie GA, Lindow SE (1995) The secret life of foliar bacterial pathogens on leaves. Annu Rev Phytopathol 33(1):145–172CrossRefPubMedGoogle Scholar
Wilson M, Hirano S, Lindow S (1999) Location and survival of leaf-associated bacteria in relation to pathogenicity and potential for growth within the leaf. Appl Environ Microbiol 65(4):1435–1443PubMedPubMedCentralGoogle Scholar
Bunster L, Fokkema NJ, Schippers B (1989) Effect of surface-active Pseudomonas spp. on leaf wettability. Appl Environ Microbiol 55(6):1340–1345PubMedPubMedCentralGoogle Scholar
Quigley NB, Gross D (1994) Syringomycin production among strains of Pseudomonas syringae pv. syringae: conservation of the syrB and syrD genes and activation of phytotoxin production by plant signal molecules. Mol Plant Microbe Interact 7:78–78CrossRefPubMedGoogle Scholar
Schroeder JI et al (2001) Guard cell signal transduction. Annu Rev Plant Biol 52(1):627–658CrossRefGoogle Scholar
Melotto M et al (2006) Plant stomata function in innate immunity against bacterial invasion. Cell 126(5):969–980CrossRefPubMedGoogle Scholar
Mittal S, Davis KR (1995) Role of the phytotoxin coronatine in the infection of Arabidopsis thaliana by Pseudomonas syringae pv. tomato. Mol Plant Microbe Interactions 8(1):165–171CrossRefGoogle Scholar
Schulze-Lefert P, Robatzek S (2006) Plant pathogens trick guard cells into opening the gates. Cell 126(5):831–834CrossRefPubMedGoogle Scholar
Gudesblat GE, Torres PS, Vojnov AA (2009) Xanthomonas campestris overcomes Arabidopsis stomatal innate immunity through a DSF cell-to-cell signal-regulated virulence factor. Plant Physiol 149(2):1017–1027CrossRefPubMedPubMedCentralGoogle Scholar
Turner NC, Graniti A (1969) Fusicoccin: a fungal toxin that opens stomata. Nature 223(5210):1070–1071CrossRefGoogle Scholar
Godoy G et al (1990) Use of mutants to demonstrate the role of oxalic acid in pathogenicity of Sclerotinia sclerotiorum on Phaseolus vulgaris. Physiol Mol Plant Pathol 37(3):179–191CrossRefGoogle Scholar
Desikan R et al (2002) A new role for an old enzyme: nitrate reductase-mediated nitric oxide generation is required for abscisic acid-induced stomatal closure in Arabidopsis thaliana. Proc Natl Acad Sci U S A 99(25):16314–16318CrossRefPubMedPubMedCentralGoogle Scholar
Zeng W, He SY (2010) A prominent role of the flagellin receptor FLAGELLIN-SENSING2 in mediating stomatal response to Pseudomonas syringae pv tomato DC3000 in Arabidopsis. Plant Physiol 153(3):1188–1198CrossRefPubMedPubMedCentralGoogle Scholar
Mustilli A-C et al (2002) Arabidopsis OST1 protein kinase mediates the regulation of stomatal aperture by abscisic acid and acts upstream of reactive oxygen species production. Plant Cell 14(12):3089–3099CrossRefPubMedPubMedCentralGoogle Scholar