Abstract
Plants are exposed to many potentially pathogenic microbes in the environment, but each species is only susceptible to a limited number of pathogens. The broad resistance is referred to as nonhost resistance. To date, little is known about the underlying mechanism of nonhost resistance and the signaling transduction process. Here we describe a simple method for isolating Arabidopsis nonhost resistance mutants against a nonadapted bacterial pathogen. A RAP2.6 promoter-driven LUC reporter system was developed to replace the tedious bacterial growth assay during the primary screening. The RAP2.6-LUC reporter gene is normally induced by the virulent bacterium Pseudomonas syringae pv tomato but not the nonadapted bacterium P. syringae pv phaseolicola. By using this method we isolated 4 mutants displaying strong reporter activity in response to P. syringae pv phaseolicola, which were characterized in some details. ebs1, ebs2, ebs3, and ebs4 (enhanced bacterial susceptibility) were compromised in resistance against P. syringae pv phaseolicola and/or P. syringae pv tomato. In addition, ebs4 showed enhanced hypersensitive response to the incompatible bacterium P. syringae pv tomato (avrB). These results demonstrated that the method is suited for large scale screening for nonhost resistance mutants.
Similar content being viewed by others
References
Heath M C. Evolution of plant resistance and susceptibility to fungal invaders. Can J Plant Pathol, 1987, 9: 389–397
Heath M C. Plant resistance to fungi. Can J Plant Pathol, 1996, 18: 469–475
Staskawicz B J, Ausubel F M, Baker B J, et al. Molecular genetics of plant disease resistance. Science, 1995, 268: 661–667
Flor H H. Current status of the gene-for-gene concept. Annu Rev Phytopathol, 1971, 9: 275–296
Lu M, Tang X, Zhou J M. Arabidopsis NHO1 is required for general resistance against Pseudomonas bacteria. Plant Cell, 2001, 13: 437–447
Li X, Lin H, Zhang W, et al. Flagellin induces innate immunity in nonhost interactions that is suppressed by Pseudomonas syringae effectors. Proc Natl Acad Sci USA, 2005, 102: 12990–12995
Assaad F F, Qiu J L, Youngs H, et al. The PEN1 syntaxin defines a novel cellular compartment upon fungal attack and is required for the timely assembly of papillae. Mol Biol Cell, 2004, 15: 5118–5129
Lipka V, Dittgen J, Bednare P, et al. Pre-and postinvasion defenses both contribute to nonhost resistance in Arabidopsis. Science, 2005, 310: 1180–1183
Stein M, Dittgen J, Sánchez-RodrÍguezc C, et al. Arabidopsis PEN3/PDR8, an ATP binding cassette transporter, contributes to nonhost resistance to inappropriate pathogens that enter by direct penetration. Plant Cell, 2006, 18: 731–746
Yun B W, Atkinson H A, Gaborit C, et al. Loss of actin cytoskeletal function and EDS1 activity, in combination, severely compromises non-host resistance in Arabidopsis against wheat powdery mildew. Plant J, 2003, 34: 768–777
Peart J R, Lu R, Sadanandom A, et al. Ubiquitin ligase-associated protein SGT1 is required for host and nonhost disease resistance in plants. Proc Natl Acad Sci USA, 2002, 99: 10865–10869
Yu G L, Katagiri F, Ausubel F M. Arabidopsis mutations at the RPS2 locus result in loss of resistance to Pseudomonas syringae strains expressing the avirulent gene avrRpt2. Mol Plant-Microbe Interact, 1993, 6: 434–443
Okamuro J K, Caster B, Villarroel R, et al. The AP2 domain of APETALA2 defines a large new family of DNA binding proteins in Arabidopsis. Proc Natl Acad Sci USA, 1997, 94: 7076–7081
He P, Chintamanani S, Chen Z, et al. Activation of a COI1-dependent pathway in Arabidopsis by Pseudomonas syringae type III effectors and coronatine. Plant J, 2004, 37: 589–602
Shang Y, Li X, Cui H, et al. RAR1, a central player in plant immunity, is targeted by Pseudomonas syringae effector AvrB. Proc Natl Acad Sci USA, 2006, 103: 19200–19205
Goldsbrough P B, Hatch E M, Huang B, et al. Gene amplification in glyphosate-tolerant tobacco cells. Plant Sci, 1990, 72: 53–62
Xiao Y, Heu S, Yi J, et al. Identification of a putative alternate sigma factor and characterization of a multicomponent regulatory cascade controlling the expression of Pseudomonas syringae pv. syringae Pss6l hrp and hrmA genes. J Bact, 1994, 176: 1025–1036
Fouts D E, Abramovitch R B, Alfano J R, et al. Genomewide identification of Pseudomonas syringae pv. tomato DC3000 promoters controlled by the HrpL alternative sigma factor. Proc Natl Acad Sci USA, 2002, 99, 2275–2280
Kamoun S. Nonhost resistance to Phytophthora: novel prospects for a classical problem. Curr Opin Plant Biol, 2001, 4: 295–300
Kazmierczak M J, Wiedmann M, Boor K J. Alternative sigma factors and their roles in bacterial virulence. Microbiol Mol Biol Rev, 2005, 69: 527–543
Bisgrove S R, Simonich M T, Smith N M, et al. A disease resistance gene in Arabidopsis with specificity for two different pathogen avirulence genes. Plant Cell, 1994, 6: 927–933
Heath M C. Nonhost resistance and nonspecific plant defenses. Curr Opin Plant Biol, 2000, 3: 315–319
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by the Shanghai Leading Academic Discipline Project (Grant No. B209) and by a grant from Chinese Ministry of Science and Technology to Zhou Jian Min (Grant No. 2003AA210080)
About this article
Cite this article
Chen, H., Pan, J., Zhao, X. et al. Reporter-based screen for Arabidopsis mutants compromised in nonhost resistance. Chin. Sci. Bull. 53, 1027–1034 (2008). https://doi.org/10.1007/s11434-008-0144-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11434-008-0144-5