A mutant CHS3 protein with TIR-NB-LRR-LIM domains modulates growth, cell death and freezing tolerance in a temperature-dependent manner in Arabidopsis.
Low temperature is one of environmental factors that restrict plant growth homeostasis and plant-pathogen interactions. Recent studies suggest a link between temperature responses and defense responses; however, the underlying molecular mechanisms remain unclear. In this study, the chilling sensitive 3 (chs3-1) mutant in Arabidopsis was characterized. chs3-1 plants showed arrested growth and chlorosis when grown at 16 degrees C or when shifted from 22 to 4 degrees C. chs3-1 plants also exhibited constitutively activated defense responses at 16 degrees C, which were alleviated at a higher temperature (22 degrees C). Map-based cloning of CHS3 revealed that it encodes an unconventional disease resistance (R) protein belonging to the TIR-NB-LRR class with a zinc-binding LIM domain (Lin-11, Isl-1 and Mec-3 domains) at the carboxyl terminus. The chs3-1 mutation in the conserved LIM-containing domain led to the constitutive activation of the TIR-NB-LRR domain. Consistently, the growth and defense phenotypes of chs3-1 plants were completely suppressed by eds1, sgt1b and rar1, partially by pad4 and nahG, but not by npr1 and ndr1. Intriguingly, chs3-1 plants grown at 16 degrees C showed enhanced tolerance to freezing temperatures. This tolerance was correlated with growth defect and cell death phenotypes caused by activated defense responses. Other mutants with activated defense responses, including cpr1, cpr5 and slh1 also displayed enhanced freezing tolerance. These findings revealed a role of an unconventional mutant R gene in plant growth, defense response and cold stress, suggesting a mutual interaction between cold signaling and defense responses.
Yang
Haibian
H
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193.
Shi
Yiting
Y
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193.
Liu
Jingyan
J
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193.
Guo
Lin
L
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193.
Zhang
Xiaoyan
X
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193.
Yang
Shuhua
S
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193.|National Plant Gene Research Center, Beijing 100193, China.
D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D016923:Cell Death; D003001:Cloning, Molecular; D003080:Cold Temperature; D018506:Gene Expression Regulation, Plant; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D054730:Protein Interaction Domains and Motifs; D018749:RNA, Plant / Q000235:genetics
D029681:Arabidopsis Proteins; D050505:Mutant Proteins; D018749:RNA, Plant
Aarts, N., Metz, M., Holub, E., Staskawicz, B.J., Daniels, M.J. and Parker, J.E. (1998) Different requirements for EDS1 and NDR1 by disease resistance genes define at least two R gene-mediated signaling pathways in Arabidopsis. Proc. Natl. Acad. Sci. USA, 95, 10306-10311.
Ade, J., DeYoung, B.J., Golstein, C. and Innes, R.W. (2007) Indirect activation of a plant nucleotide binding site-leucine-rich repeat protein by a bacterial protease. Proc. Natl. Acad. Sci. USA, 104, 2531-2536.
Agarwal, M., Hao, Y., Kapoor, A., Dong, C.H., Fujii, H., Zheng, X. and Zhu, J.K. (2006) A R2R3 type MYB transcription factor is involved in the cold regulation of CBF genes and in acquired freezing tolerance. J. Biol. Chem. 281, 37636-37645.
Alcazar, R., Garcia, A.V., Parker, J.E. and Reymond, M. (2009) Incremental steps toward incompatibility revealed by Arabidopsis epistatic interactions modulating salicylic acid pathway activation. Proc. Natl. Acad. Sci. USA, 106, 334-339.
Arber, S. and Caroni, P. (1996) Specificity of single LIM motifs in targeting and LIM/LIM interactions in situ. Genes Dev. 10, 289-300.
Austin, M.J., Muskett, P., Kahn, K., Feys, B.J., Jones, J.D. and Parker, J.E. (2002) Regulatory role of SGT1 in early R gene-mediated plant defenses. Science, 295, 2077-2080.
Azevedo, C., Betsuyaku, S., Peart, J., Takahashi, A., Noel, L., Sadanandom, A., Casais, C., Parker, J. and Shirasu, K. (2006) Role of SGT1 in resistance protein accumulation in plant immunity. EMBO J. 25, 2007-2016.
Bates, L.S., Waldren, R.P. and Teare, I.D. (1972) Rapid determination of free proline for water-stress studies. Plant Soil, 39, 205-207.
Bendahmane, A., Farnham, G., Moffett, P. and Baulcombe, D.C. (2002) Constitutive gain-of-function mutants in a nucleotide binding site-leucine rich repeat protein encoded at the Rx locus of potato. Plant J. 32, 195-204.
Bieri, S., Mauch, S., Shen, Q.H. et al. (2004) RAR1 positively controls steady state levels of barley MLA resistance proteins and enables sufficient MLA6 accumulation for effective resistance. Plant Cell, 16, 3480-3495.
Bomblies, K., Lempe, J., Epple, P., Warthmann, N., Lanz, C., Dangl, J.L. and Weigel, D. (2007) Autoimmune response as a mechanism for a Dobzhansky-Muller-type incompatibility syndrome in plants. PLoS Biol. 5, e236.
Bowling, S.A., Guo, A., Cao, H., Gordon, A.S., Klessig, D.F. and Dong, X. (1994) A mutation in Arabidopsis that leads to constitutive expression of systemic acquired resistance. Plant Cell, 6, 1845-1857.
Bowling, S.A., Clarke, J.D., Liu, Y., Klessig, D.F. and Dong, X. (1997) The cpr5 mutant of Arabidopsis expresses both NPR1-dependent and NPR1-independent resistance. Plant Cell, 9, 1573-1584.
Chinnusamy, V., Ohta, M., Kanrar, S., Lee, B.H., Hong, X., Agarwal, M. and Zhu, J.K. (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev. 17, 1043-1054.
Chinnusamy, V., Zhu, J. and Zhu, J.K. (2007) Cold stress regulation of gene expression in plants. Trends Plant Sci. 12, 444-451.
Clough, S.J. and Bent, A.F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16, 735-743.
Dangl, J.L. and Jones, J.D. (2001) Plant pathogens and integrated defence responses to infection. Nature, 411, 826-833.
Doherty, C.J., Van Buskirk, H.A., Myers, S.J. and Thomashow, M.F. (2009) Roles for Arabidopsis CAMTA transcription factors in cold-regulated gene expression and freezing tolerance. Plant Cell, 21, 972-984.
Du, L., Ali, G.S., Simons, K.A., Hou, J., Yang, T., Reddy, A.S. and Poovaiah, B.W. (2009) Ca2+/calmodulin regulates salicylic-acid-mediated plant immunity. Nature, 457, 1154-1158.
Durrant, W.E. and Dong, X. (2004) Systemic acquired resistance. Annu. Rev. Phytopathol. 42, 185-209.
Feuerstein, R., Wang, X., Song, D., Cooke, N.E. and Liebhaber, S.A. (1994) The LIM/double zinc-finger motif functions as a protein dimerization domain. Proc. Natl. Acad. Sci. USA, 91, 10655-10659.
Freyd, G., Kim, S.K. and Horvitz, H.R. (1990) Novel cysteine-rich motif and homeodomain in the product of the Caenorhabditis elegans cell lineage gene lin-11. Nature, 344, 876-879.
Gao, M., Liu, J., Bi, D., Zhang, Z., Cheng, F., Chen, S. and Zhang, Y. (2008) MEKK1, MKK1/MKK2 and MPK4 function together in a mitogen-activated protein kinase cascade to regulate innate immunity in plants. Cell Res. 18, 1190-1198.
Garrett, K.A., Dendy, S.P., Frank, E.E., Rouse, M.N. and Travers, S.E. (2006) Climate change effects on plant disease: genomes to ecosystems. Annu Rev Phytopathol. 44, 489-509.
Gilmour, S.J., Artus, N.N. and Thomashow, M.F. (1992) cDNA sequence analysis and expression of two cold-regulated genes of Arabidopsis thaliana. Plant Mol. Biol. 18, 13-21.
Glazebrook, J. (2001) Genes controlling expression of defense responses in Arabidopsis--2001 status. Curr. Opin. Plant Biol. 4, 301-308.
Glazebrook, J., Rogers, E.E. and Ausubel, F.M. (1996) Isolation of Arabidopsis mutants with enhanced disease susceptibility by direct screening. Genetics, 143, 973-982.
Gou, M., Su, N., Zheng, J., Huai, J., Wu, G., Zhao, J., He, J., Tang, D., Yang, S. and Wang, G. (2009) An F-box gene, CPR30, functions as a negative regulator of the defense response in Arabidopsis. Plant J. 60, 757-770.
Hellens, R.P., Edwards, E.A., Leyland, N.R., Bean, S. and Mullineaux, P.M. (2000) pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation. Plant Mol. Biol. 42, 819-832.
Hua, J. (2009) From freezing to scorching, transcriptional responses to temperature variations in plants. Curr. Opin. Plant Biol. 12, 568-573.
Hua, J., Grisafi, P., Cheng, S.H. and Fink, G.R. (2001) Plant growth homeostasis is controlled by the Arabidopsis BON1 and BAP1 genes. Genes Dev. 15, 2263-2272.
Hwang, C.F., Bhakta, A.V., Truesdell, G.M., Pudlo, W.M. and Williamson, V.M. (2000) Evidence for a role of the N terminus and leucine-rich repeat region of the Mi gene product in regulation of localized cell death. Plant Cell, 12, 1319-1329.
Ichimura, K., Casais, C., Peck, S.C., Shinozaki, K. and Shirasu, K. (2006) MEKK1 is required for MPK4 activation and regulates tissue-specific and temperature-dependent cell death in Arabidopsis. J. Biol. Chem. 281, 36969-36976.
Kadrmas, J.L. and Beckerle, M.C. (2004) The LIM domain: from the cytoskeleton to the nucleus. Nat. Rev. Mol. Cell Biol. 5, 920-931.
Lee, H., Guo, Y., Ohta, M., Xiong, L., Stevenson, B. and Zhu, J.K. (2002) LOS2, a genetic locus required for cold-responsive gene transcription encodes a bi-functional enolase. EMBO J. 21, 2692-2702.
Lee, J., Nam, J., Park, H.C. et al. (2007) Salicylic acid-mediated innate immunity in Arabidopsis is regulated by SIZ1 SUMO E3 ligase. Plant J. 49, 79-90.
Leister, R.T., Dahlbeck, D., Day, B., Li, Y., Chesnokova, O. and Staskawicz, B.J. (2005) Molecular genetic evidence for the role of SGT1 in the intramolecular complementation of Bs2 protein activity in Nicotiana benthamiana. Plant Cell, 17, 1268-1278.
Li, X., Zhang, Y., Clarke, J.D., Li, Y. and Dong, X. (1999) Identification and cloning of a negative regulator of systemic acquired resistance, SNI1, through a screen for suppressors of npr1-1. Cell, 98, 329-339.
Li, X., Clarke, J.D., Zhang, Y. and Dong, X. (2001) Activation of an EDS1-mediated R-gene pathway in the snc1 mutant leads to constitutive, NPR1-independent pathogen resistance. Mol. Plant Microbe Interact. 14, 1131-1139.
Li, Y., Zheng, L., Corke, F., Smith, C. and Bevan, M.W. (2008) Control of final seed and organ size by the DA1 gene family in Arabidopsis thaliana. Genes Dev. 22, 1331-1336.
Liu, Y., Schiff, M., Marathe, R. and Dinesh-Kumar, S.P. (2002) Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J. 30, 415-429.
Meyers, B.C., Kozik, A., Griego, A., Kuang, H. and Michelmore, R.W. (2003) Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant Cell, 15, 809-834.
Miura, K. and Ohta, M. (2009) SIZ1, a small ubiquitin-related modifier ligase, controls cold signaling through regulation of salicylic acid accumulation. J. Plant Physiol. 167, 555-560.
Miura, K., Jin, J.B., Lee, J., Yoo, C.Y., Stirm, V., Miura, T., Ashworth, E.N., Bressan, R.A., Yun, D.J. and Hasegawa, P.M. (2007) SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis. Plant Cell, 19, 1403-1414.
Muskett, P.R., Kahn, K., Austin, M.J., Moisan, L.J., Sadanandom, A., Shirasu, K., Jones, J.D. and Parker, J.E. (2002) Arabidopsis RAR1 exerts rate-limiting control of R gene-mediated defenses against multiple pathogens. Plant Cell, 14, 979-992.
Nawrath, C. and Metraux, J.P. (1999) Salicylic acid induction-deficient mutants of Arabidopsis express PR-2 and PR-5 and accumulate high levels of camalexin after pathogen inoculation. Plant Cell, 11, 1393-1404.
Noutoshi, Y., Ito, T., Seki, M., Nakashita, H., Yoshida, S., Marco, Y., Shirasu, K. and Shinozaki, K. (2005) A single amino acid insertion in the WRKY domain of the Arabidopsis TIR-NBS-LRR-WRKY-type disease resistance protein SLH1 (sensitive to low humidity 1) causes activation of defense responses and hypersensitive cell death. Plant J. 43, 873-888.
Qiu, J.L., Zhou, L., Yun, B.W., Nielsen, H.B., Fiil, B.K., Petersen, K., Mackinlay, J., Loake, G.J., Mundy, J. and Morris, P.C. (2008) Arabidopsis mitogen-activated protein kinase kinases MKK1 and MKK2 have overlapping functions in defense signaling mediated by MEKK1, MPK4, and MKS1. Plant Physiol. 148, 212-222.
Schmeichel, K.L. and Beckerle, M.C. (1994) The LIM domain is a modular protein-binding interface. Cell, 79, 211-219.
Schneider, J.C., Hugly, S. and Somerville, C.R. (1995) Chilling-sensitive mutants of Arabidopsis. Plant Mol. Biol. Rep. 13, 11-17.
Seo, P.J., Kim, M.J., Park, J.Y., Kim, S.Y., Jeon, J., Lee, Y.H., Kim, J. and Park, C.M. (2009) Cold activation of a plasma membrane-tethered NAC transcription factor induces a pathogen resistance response in Arabidopsis. Plant J. 61, 661-671.
Shirasu, K., Lahaye, T., Tan, M.W., Zhou, F., Azevedo, C. and Schulze-Lefert, P. (1999) A novel class of eukaryotic zinc-binding proteins is required for disease resistance signaling in barley and development in C. elegans. Cell, 99, 355-366.
Someya, N., Niinuma, K., Kimura, M., Yamaguchi, I. and Hamamoto, H. (2004) Pattern of N gene-mediated systemic hypersensitive response and turnover of viral replicase protein in tobacco. Arch. Virol. 149, 2105-2113.
Tao, Y., Yuan, F., Leister, R.T., Ausubel, F.M. and Katagiri, F. (2000) Mutational analysis of the Arabidopsis nucleotide binding site-leucine-rich repeat resistance gene RPS2. Plant Cell, 12, 2541-2554.
Thomas, C., Hoffmann, C., Gatti, S. and Steinmetz, A. (2007) LIM Proteins: a novel nlass of actin cytoskeleton organizers in plants. Plant Signal Behav. 2, 99-100.
Thomashow, M.F. (1999) PLANT COLD ACCLIMATION: freezing tolerance genes and regulatory mechanisms. Annu. Rev. Plant Physiol. Plant Mol. Biol. 50, 571-599.
Thordal-Christensen, H., Zhang, Z., Wei, Y. and Collinge, D.B. (1997) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J. 11, 1187-1194.
Tornero, P., Merritt, P., Sadanandom, A., Shirasu, K., Innes, R.W. and Dangl, J.L. (2002) RAR1 and NDR1 contribute quantitatively to disease resistance in Arabidopsis, and their relative contributions are dependent on the R gene assayed. Plant Cell, 14, 1005-1015.
Tsutsui, T., Kato, W., Asada, Y. et al. (2009) DEAR1, a transcriptional repressor of DREB protein that mediates plant defense and freezing stress responses in Arabidopsis. J. Plant. Res. 122, 633-643.
Wang, Y., Bao, Z., Zhu, Y. and Hua, J. (2009) Analysis of temperature modulation of plant defense against biotrophic microbes. Mol. Plant Microbe Interact., 22, 498-506.
Wildermuth, M.C., Dewdney, J., Wu, G. and Ausubel, F.M. (2001) Isochorismate synthase is required to synthesize salicylic acid for plant defence. Nature, 414, 562-565.
Xiao, S., Charoenwattana, P., Holcombe, L. and Turner, J.G. (2003) The Arabidopsis genes RPW8.1 and RPW8.2 confer induced resistance to powdery mildew diseases in tobacco. Mol. Plant Microbe Interact. 16, 289-294.
Xin, Z. and Browse, J. (1998) Eskimo1 mutants of Arabidopsis are constitutively freezing-tolerant. Proc. Natl. Acad. Sci. USA, 95, 7799-7804.
Yang, S. and Hua, J. (2004) A haplotype-specific Resistance gene regulated by BONZAI1 mediates temperature-dependent growth control in Arabidopsis. Plant Cell, 16, 1060-1071.
Yang, H., Li, Y. and Hua, J. (2006) The C2 domain protein BAP1 negatively regulates defense responses in Arabidopsis. Plant J. 48, 238-248.
Yoo, C.Y., Miura, K., Jin, J.B., Lee, J., Park, H.C., Salt, D.E., Yun, D.J., Bressan, R.A. and Hasegawa, P.M. (2006) SIZ1 small ubiquitin-like modifier E3 ligase facilitates basal thermotolerance in Arabidopsis independent of salicylic acid. Plant Physiol. 142, 1548-1558.
Zhou, F., Mosher, S., Tian, M., Sassi, G., Parker, J. and Klessig, D.F. (2008) The Arabidopsis gain-of-function mutant ssi4 requires RAR1 and SGT1b differentially for defense activation and morphological alterations. Mol. Plant Microbe Interact. 21, 40-49.
Zhu, J., Verslues, P.E., Zheng, X. et al. (2005) HOS10 encodes an R2R3-type MYB transcription factor essential for cold acclimation in plants. Proc. Natl. Acad. Sci. USA, 102, 9966-9971.
Zhu, J., Jeong, J.C., Zhu, Y. et al. (2008) Involvement of Arabidopsis HOS15 in histone deacetylation and cold tolerance. Proc. Natl. Acad. Sci. USA, 105, 4945-4950.