Skip to main content
SpringerLink
Log in

Molecular characterization of a pepper C2 domain-containing SRC2 protein implicated in resistance against host and non-host pathogens and abiotic stresses

  • Rapid Communication
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Plants guard themselves against pathogen attack using multi-layered defense mechanism. Calcium represents an important secondary messenger during such defense responses. Upon examination of a pepper cDNA library, we observed that the gene CaSRC2-1 (Capsicum annum SRC2-1) was upregulated significantly in response to infection with the type II non-host pathogen Xanthomonas axonopodis pv. glycines 8 ra, which elicits a hypersensitive response. CaSRC2-1 encodes a protein that contains a C2 domain and it exhibits a high degree of homology to the protein Soybean genes regulated by cold 2 (SRC2). However, little is known about how SRC2 expression is elicited by biotic stresses such as pathogen challenge. Further sequence analysis indicated that the CaSRC2-1 C2 domain is unique and contain certain amino acids that are conserved within the C2 domains of other plants and animals. CaSRC2-1 transcription was up-regulated under both biotic and abiotic stress conditions, including bacterial and viral pathogen infection, CaCl2 and cold treatment, but unaffected by treatment with plant defense-related chemicals such as salicylic acid, methyl jasmonic acid, ethephone, and abscisic acid. Intriguingly, under steady state conditions, CaSRC2-1 was expressed only in the root system. A CaSRC2-1-GFP fusion protein was used to determine localization to the plasma membrane. A fusion protein lacking the C2 domain failed to target the membrane but remained in the cytoplasm, indicating that the C2 domain plays a critical role in localization. Thus, CaSRC2-1 encodes a novel C2 domain-containing protein that targets the plasma membrane and plays a critical role in the abiotic stress and defense responses of pepper plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Abbreviations

HR:

Hypersensitive response

CaSRC2:

Capsicum annum soybean genes regulated by cold 2

References

  • Abel S, Theologis A (1994) Transient transformation of Arabidopsis leaf protoplasts: a versatile experimental system to study gene expression. Plant J 5:421–427

    Article  PubMed  CAS  Google Scholar 

  • Aubourg S, Picaud A, Kreis M, Lecharny A (1999) Structure and expression of three SRC2 homologues and a novel subfamily of flavoprotein monooxygenase genes revealed by the analysis of a 25 kb fragment from Arabidopsis thaliana chromosome IV. Gene 230:197–205

    Article  PubMed  CAS  Google Scholar 

  • Baker B, Zambryski P, Staskawicz B, Dinesh-Kumar SP (1997) Signaling in Plant-Microbe Interactions. Science 276:726–733

    Article  PubMed  CAS  Google Scholar 

  • Benes CH, Wu N, Elia AE, Dharia T, Cantley LC, Soltoff SP (2005) The C2 domain of PKCδ is a phosphotyrosine binding domain. Cell 121:271–280

    Article  PubMed  CAS  Google Scholar 

  • Choi D, Kim HM, Yun HK, Park JA, Kim WT, Bok SH (1996) Molecular cloning of a methallothionein-like gene from Nicotiana glutinosa L. and its induction by wounding and tobacco mosaic virus infection. Plant Physiol 112:353–359

    Article  PubMed  CAS  Google Scholar 

  • Chung E, Kim SY, Yi SY, Choi D (2003) Capsicum annuum dehydrin, an osmotic-stress gene in hot pepper plants. Mol Cells 15:327–332

    PubMed  CAS  Google Scholar 

  • Clark JD, Lin LL, Kriz RW, Ramesha CS, Sultzman LA, Lin AY, Milona N, Knopf JL (1991) A novel arachidonic acid selective cytosolic PLA2 contains a Ca2+dependent translocation domain with homology to PKC and GAP. Cell 65:1043–1051

    Article  PubMed  CAS  Google Scholar 

  • Chung E, Ryu C-M, Oh S-K, Kim RN, Park JM, Cho HS, Lee S, Moon JS, Park S-H, Choi D (2006) Suppression of pepper SGT1 and SKP1 causes severe retardation of plant growth and compromises basal resistance. Physiol Plant 126:605–617

    CAS  Google Scholar 

  • Dangl JL, Dietrich RA, Richberg MH (1996) Death don’t have no mercy: Cell death programs in plant-microbe interactions. Plant Cell 8:1793–1807

    Article  PubMed  CAS  Google Scholar 

  • David SJ, Viestra RD (1996) Soluble derivatives of green fluorescent protein (GFP) for use in Arabidopsis thaliana. Weeds World 3:43–48

    Google Scholar 

  • Dong X (1998) SA, JA, ethylene, and disease resistance in plants. Curr Opin Plant Biol 1:316–323

    Article  PubMed  CAS  Google Scholar 

  • Gerke V, Moss SE (2002) Annexins: from structure to function. Physiol Rev 82:331–371

    PubMed  CAS  Google Scholar 

  • Heath MC (2000) Non-host resistance and nonspecific plant defenses. Curr Opin Plant Biol 3:315–319

    Article  PubMed  CAS  Google Scholar 

  • Hetherington AM, Brownlee C (2004) The generation of Ca2+ signals in plants. Annu Rev Plant Biol 55:401–427

    Article  PubMed  CAS  Google Scholar 

  • Kaibuchi K, Fukumoto Y, Oku N, Takai Y, Arai K, Muramatsu M (1989) Molecular genetic analysis of the regulatory and catalytic domains of protein kinase C. J Biol Chem 264:13489–13496

    PubMed  CAS  Google Scholar 

  • Kamoun S (2001) Non-host resistance to Phytophthora: novel prospects for a classical problem. Curr Opin Plant Biol 4:295–300

    Article  PubMed  CAS  Google Scholar 

  • Kim CY, Koo YD, Jin JB, Moo BC, Kang CH, Kim ST, Park BO, Lee SY, Kim ML, Hwang I, Kang KY, Bahk JD, Lee SY, Cho MJ (2003) Rice C2-domain proteins are induced and translocated to the plasma membrane in response to a fungal elicitor. Biochemistry 42:11625–11633

    Article  PubMed  CAS  Google Scholar 

  • Kim YJ, Kim JE, Lee JH, Lee MH, Jung HW, Bahk YY, Hwang BK, Hwang I, Kim WT (2004) The Vr-PLC3 gene encodes a putative plasma membrane-localized phosphoinositide-specific phospholipase C whose expression is induced by abiotic stress in mung bean (Vigna radiata L.). FEBS Lett 556:127–136

    Article  PubMed  CAS  Google Scholar 

  • Kopka J, Pical C, Hetherington AM, Müller-Röber B (1998) Ca2+/phospholipids-binding (C2) domain in multiple plant proteins: novel components of the calcium-sensing apparatus. Plant Mol Biol 36:627–637

    Article  PubMed  CAS  Google Scholar 

  • Kretsinger RH (1980) Structure and evolution of calcium-modulated proteins. Crit Rev Biochem 8:119–174

    Article  CAS  Google Scholar 

  • Lee S, Kim SY, Chung E, Joung YH, Pai HS, Hur CG, Choi D (2004) EST and microarray analyses of pathogen-responsive genes in hot pepper (Capsicum annuum L.) non-host resistance against soybean pustule pathogen (Xanthomonas axonopodis pv. glycines). Funct Integr Genomics 4:196–205

    Article  PubMed  Google Scholar 

  • Mauchi-Mani B, Mauch F (2005) The role of abscisic acid in plant-pathogen interactions. Curr Opin Plant Biol 8:409–414

    Article  CAS  Google Scholar 

  • Miyazaki M, Kaibuchi K, Shirataki H, Kohno H, Ueyama T, Nishikawa J, Takai Y (1995) Rabphilin3A binds to a M(r) 115,000 polypeptide in a phosphatidylserineand Ca2+dependent manner. Brain Res Mol Brain Res 28:29–36

    Article  PubMed  CAS  Google Scholar 

  • Mysore KS, Ryu C-M (2004) Non-host resistance: how much do we know? Trends Plant Sci 9:97–104

    Article  PubMed  CAS  Google Scholar 

  • Nürnberger T, Brunner F, Kemmerling B, Piater L (2004) Innate immunity in plants and animals: striking similarities and obvious differences. Immunol Rev 198:249–266

    Article  PubMed  Google Scholar 

  • Nalefski EA, Falke JJ (1996) The C2 domain calcium-binding motif: structural and functional diversity. Protein Sci 5:2375–2390

    Article  PubMed  CAS  Google Scholar 

  • Oh S-K, Park JM, Joung YH, Lee S, Chung E, Kim S-Y, Yu SH, Choi D (2005) A plant EPF-type zinc-finger protein, CaPF1, involved in defence against pathogens. Mol Plant Pathol 6:269–285

    Article  CAS  Google Scholar 

  • Oh S-K, Lee S, Chung E, Park JM, Yu SH, Ryu C-M, Choi D (2006) Insight into Types I and II non-host resistance using expression patterns of defense-related genes in tobacco. Planta 213:1102–1107

    Google Scholar 

  • Oufattole M, Park JH, Poxleitner M, Jiang L, Rogers JC (2005) Selective membrane protein internalization accompanies movement from the endoplasmic reticulum to the protein storage vacuole pathway in Arabidopsis. Plant Cell 17:3066–3080

    Article  PubMed  CAS  Google Scholar 

  • Park JM, Park CJ, Lee SB, Ham BK, Shin R, Paek KH (2001) Overexpression of the tobacco Tsi gene encoding an EREBP/AP2-type transcription factor enhances resistance against pathogen attack and osmotic stress in tobacco. Plant Cell 13:1035–1046

    Article  PubMed  CAS  Google Scholar 

  • Pepio AM, Sossin WS (2001) Membrane translocation of novel protein kinase Cs is regulated by phosphorylation of the C2 domain. J Biol Chem 276:3846–3855

    Article  PubMed  CAS  Google Scholar 

  • Reymond P, Farmer EE (1998) Jasmonate and salicylate as global signals for defense gene expression. Curr Opin Plant Biol 1:404–411

    Article  PubMed  CAS  Google Scholar 

  • Rizo J, Südhof TC (1998) C2-domains, structure and function of a universal Ca2+-binding domain. J Biol Chem 273:15879–15882

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3nd. Cold Spring Harbor Laboratory, Cold Spring Harbor, Cold Springer Harbor

    Google Scholar 

  • Shin R, Lee GJ, Park CJ, Kim TY, You JS, Nam YW, Paek KH (2001) Isolation of pepper mRNAs differentially expressed during the hypersensitive response to tobacco mosaic virus and characterization of a proteinase inhibitor gene. Plant Sci 161:727–737

    Article  CAS  Google Scholar 

  • Ryu C-M, Anand A, Kang L, Mysore KS (2004) Agrodrench: a novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse Solanaceous species. Plant J 40:322–331

    Article  PubMed  CAS  Google Scholar 

  • Suh MC, Oh S-K, Kim Y-C, Pai H-S, Choi D (2003) Expression of a novel tobacco gene, NgCDM1, is preferentially associated with pathogen-induced cell death. Physiol Mol Plant Pathol 6:227–235

    Article  CAS  Google Scholar 

  • Takahashi R, Shimosaka E (1997) cDNA sequence analysis and expression of two cold-regulated genes in soybean. Plant Sci 123:93–104

    Article  CAS  Google Scholar 

  • Thomma BP, Penninckx IA, Broekaert WF, Cammue BP (2001) The complexity of disease signaling in Arabidopsis. Curr Opin Plant Immun 13:63–68

    Article  CAS  Google Scholar 

  • Thordal-Christensen H (2003) Fresh insights into processes of non-host resistance. Curr Opin Plant Biol 6:351–357

    Article  PubMed  CAS  Google Scholar 

  • Wang X (2002) Phospholipase D in hormonal and stress signaling. Curr Opin Plant Biol 5:408–414

    Article  PubMed  CAS  Google Scholar 

  • Xoconostle-Cazares B, Xiang Y, Ruiz-Medrano R, Wang HL, Monzer J, Yoo BC, McFarland KC, Franceschi VR, Lucas WJ (1999) Plant paralog to viral movement protein that potentiates transport of mRNA into the phloem. Science 283:94–98

    Article  PubMed  CAS  Google Scholar 

  • Yang H, Li Y, Hua J (2006a) The C2 domain protein BAP1 negatively regulates defense responses in Arabidopsis. Plant J 48:238–248

    Article  PubMed  CAS  Google Scholar 

  • Yang S, Yang H, Grisafi P, Sanchatjate S, Fink GR, Sun Q, Hua J (2006b) The BON/CPN gene family represses cell death and promotes cell growth in Arabidopsis. Plant J 45:166–179

    Article  PubMed  CAS  Google Scholar 

  • Yang H, Yang S, Li Y, Hua J (2007) The Arabidopsis BAP1 and BAP2 genes are general inhibitors of programmed cell death. Plant Physiol 145:135–146

    Article  PubMed  CAS  Google Scholar 

  • Yi SY, Kim JH, Joung YH, Lee S, Kim WT, Yu SH, Choi D (2004) The pepper transcription factor CaPF1 confers pathogen and freezing tolerance in Arabidopsis. Plant Physiol 136:2862–2874

    Article  PubMed  CAS  Google Scholar 

  • Zhao J, Davis LC, Verpoorte R (2005) Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 23:283–333

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Ingyu Hwang for providing the bacterial strain. This research was supported by grants from the Plant Diversity Research Center (PDRC) funded by Ministry of Science and Technology (MOST), KRIBB initiative program, and a grant (#20070401034005) from BioGreen21 program, Rural Development Administration, Republic of Korea.

Author information

Authors and Affiliations

  1. Plant Genome Research Center, KRIBB, Daejeon, 305-803, South Korea

    Young-Cheol Kim, Soo-Yong Kim, Doil Choi & Jeong Mee Park

  2. Systems Microbiology Research Center, KRIBB, Daejeon, 305-803, South Korea

    Choong-Min Ryu

  3. Department of Plant Sciences, Seoul National University, Seoul, 151-742, South Korea

    Doil Choi

Authors
  1. Young-Cheol Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Soo-Yong Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Doil Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Choong-Min Ryu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jeong Mee Park
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Choong-Min Ryu or Jeong Mee Park.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, YC., Kim, SY., Choi, D. et al. Molecular characterization of a pepper C2 domain-containing SRC2 protein implicated in resistance against host and non-host pathogens and abiotic stresses. Planta 227, 1169–1179 (2008). https://doi.org/10.1007/s00425-007-0680-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-007-0680-2

Keywords

Search

Navigation