Skip to main content

Advertisement

SpringerLink
Log in

SuperSAGE revealed different classes of early resistance response genes in Capsicum chinense plants harboring L 3-resistance gene infected with Pepper mild mottle virus

  • Viral and Viroid Diseases
  • Published:
Journal of General Plant Pathology Aims and scope Submit manuscript

Abstract

We used SuperSAGE, an improved version of serial analysis of gene expression, to explore transcriptome changes early in the L 3-mediated resistance response of pepper plants against a tobamovirus. Capsicum chinense plants homozygous for the L 3 resistance gene were infected with virulent and avirulent strains of Pepper mild mottle virus (PMMoV). Plants were maintained at a temperature nonpermissive for the resistance gene to allow the viruses to spread, then transferred to a permissive temperature for 3 h and subsequently analyzed. In the incompatible reaction, we selected 152 SuperSAGE tags (each 26 nucleotides long) possibly corresponding to upregulated genes, and 84 tags for downregulated genes. Approximately 70% of tags had matching ESTs in the genus Capsicum, other genera within the Solanaceae and/or other families of plants. More than 90% of tags with EST matches could be annotated with either functionally characterized or uncharacterized proteins. We compared genes annotated by SuperSAGE tags and those annotated by partial cDNA that was obtained using the SuperSAGE tag sequences as rapid amplification of the cDNA ends-PCR primers. Of genes annotated by SuperSAGE tags, c. 90% were consistent with those annotated by longer cDNA sequences. We cloned 17 full-length cDNAs from different SuperSAGE tags and confirmed that these genes were upregulated during normal infection in the incompatible interaction. We identified several early resistance response genes including a Ran/TC4 protein and a β-oxidation multifunctional protein, indicating that SuperSAGE is a powerful tool for investigating plant–pathogen interactions.

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

Similar content being viewed by others

References

  • Ahn IP, Kim S, Lee YH (2005) Vitamin B1 functions as an activator of plant disease resistance. Plant Physiol 138:1505–1515

    Article  PubMed  CAS  Google Scholar 

  • Ahn IP, Kim S, Lee YH, Suh SC (2007) Vitamin B1-induced priming is dependent on hydrogen peroxide and the NPR1 gene in Arabidopsis. Plant Physiol 143:838–848

    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 

  • Berzal-Herranz A, de la Cruz A, Tenllado F, Díaz-Ruiz JR, López L, Sanz AI, Vaquero C, Serra MT, García-Luque I (1995) The Capsicum L 3 gene-mediated resistance against the tobamoviruses is elicited by the coat protein. Virology 209:498–505

    Article  PubMed  CAS  Google Scholar 

  • Bol JF, Linthorst HJM, Cornelissen BJC (1990) Plant pathogenesis-related proteins induced by virus infection. Annu Rev Phytopathol 28:113–138

    Article  CAS  Google Scholar 

  • Boukema IW (1982) Resistance to a new strain of TMV in Capsicum chacoense Hunz. Capsicum Newsl 1:49–51

    Google Scholar 

  • Boukema IW (1984) Resistance to TMV in Capsicum chacoense Hunz. is governed by allele of the L-locus. Capsicum Newsl 3:47–48

    Google Scholar 

  • Coemans B, Matsumura H, Terauchi R, Remy S, Swennen R, Sági L (2005) SuperSAGE combined with PCR walking allows global gene expression profiling of banana (Musa acuminata), a non-model organism. Theor Appl Genet 111:1118–1126

    Article  PubMed  CAS  Google Scholar 

  • Dellagi A, Birch PRJ, Heilbronn J, Avrova AO, Montesano M, Palva ET, Lyon GD (2000) A potato gene, erg–1, is rapidly induced by Erwinia carotovora ssp. atroseptica, Phytophthora infestans, ethylene and salicylic acid. J Plant Physiol 157:201–205

    CAS  Google Scholar 

  • Eastmond PJ, Graham IA (2000) The multifunctional protein AtMFP2 is co-ordinately expressed with other genes of fatty acid β-oxidation during seed germination in Arabidopsis thaliana (L.) Heynh. Biochem Soc Trans 28:95–99

    Article  PubMed  CAS  Google Scholar 

  • Farmer EE (2000) Adding injury to insult: pathogen detection and responses. Genome Biol 1:1012.1–1012.3

    Google Scholar 

  • García-Luque I, Ferrero ML, Rodríguez JM, Alonso E, de la Cruz A, Sanz AI, Vaquero C, Serra MT, Díaz-Ruíz JR (1993) The nucleotide sequence of the coat protein genes and 3′ non-coding regions of two resistance-breaking tobamoviruses in pepper shows that they are different viruses. Arch Virol 131:75–88

    Article  PubMed  Google Scholar 

  • Gilardi P, García-Luque I, Serra MT (2004) The coat protein of tobamovirus acts as elicitor of both L 2 and L 4 gene-mediated resistance in Capsicum. J Gen Virol 85:2077–2085

    Article  PubMed  CAS  Google Scholar 

  • Gilroy EM, Hein I, van der Hoorn R, Boevink PC, Venter E, McLellan H, Kaffarnik F, Hrubikova K, Shaw J, Holeva M, López EC, Borras-Hidalgo O, Pritchard L, Loake GJ, Lacomme C, Birch PRJ (2007) Involvement of cathepsin B in the plant disease resistance hypersensitive response. Plant J 52:1–13

    Article  PubMed  CAS  Google Scholar 

  • Goritschnig S, Zhang Y, Li X (2007) The ubiquitin pathway is required for innate immunity in Arabidopsis. Plant J 49:540–551

    Article  PubMed  CAS  Google Scholar 

  • Hamada H, Takeuchi S, Morita Y, Sawada H, Kiba A, Hikichi Y (2002) Amino acid changes in Pepper mild mottle virus coat protein that affect L 3 gene-mediated resistance in pepper. J Gen Plant Pathol 68:155–162

    Article  CAS  Google Scholar 

  • Hamada H, Takeuchi S, Kiba A, Tsuda S, Suzuki K, Hikichi Y, Okuno T (2005) Timing and extent of hypersensitive response are critical to restrict local and systemic spread of Pepper mild mottle virus in pepper containing the L 3 gene. J Gen Plant Pathol 71:90–94

    Article  CAS  Google Scholar 

  • Hamada H, Tomita R, Iwadate Y, Kobayashi K, Munemura I, Takeuchi S, Hikichi Y, Suzuki K (2007) Cooperative effect of two amino acid mutations in the coat protein of Pepper mild mottle virus overcomes L 3-mediated resistance in Capsicum plants. Virus Genes 34:205–214

    Article  PubMed  CAS  Google Scholar 

  • Hatsugai N, Kuroyanagi M, Yamada K, Meshi T, Tsuda S, Kondo M, Nishimura M, Hara-Nishimura I (2004) A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death. Science 305:855–858

    Article  PubMed  CAS  Google Scholar 

  • Joseph J (2006) Ran at a glance. J Cell Sci 119:3481–3484

    Article  PubMed  CAS  Google Scholar 

  • Kiba A, Takata O, Ohnishi K, Hikichi Y (2006) Comparative analysis of induction pattern of programmed cell death and defense-related responses during hypersensitive cell death and development of bacterial necrotic leaf spots in eggplant. Planta 224:981–994

    Article  PubMed  CAS  Google Scholar 

  • Kim CY, Koo YD, Jin JB, Moon 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 

  • Kobayashi K, Tsuge S, Stavolone L, Hohn T (2002) The cauliflower mosaic virus virion-associated protein is dispensable for viral replication in single cells. J Virol 76:9457–9464

    Article  PubMed  CAS  Google Scholar 

  • Kruger J, Thomas CM, Golstein C, Dixon MS, Smoker M, Tang S, Mulder L, Jones JD (2002) A tomato cysteine protease required for Cf-2-dependent disease resistance and suppression of autonecrosis. Science 296:744–747

    Article  PubMed  Google Scholar 

  • Matsumura H, Reich S, Ito A, Saitoh H, Kamoun S, Winter P, Kahl G, Reuter M, Kruger DH, Terauchi R (2003) Gene expression analysis of plant host–pathogen interactions by SuperSAGE. Proc Natl Acad Sci U S A 100:15718–15723

    Article  PubMed  CAS  Google Scholar 

  • Matsumura H, Ito A, Saitoh H, Winter P, Kahl G, Reuter M, Kruger DH, Terauchi R (2005) SuperSAGE. Cell Microbiol 7:11–18

    Article  PubMed  CAS  Google Scholar 

  • Moeder W, Del Pozo O, Navarre DA, Martin GB, Klessig DF (2007) Aconitase plays a role in regulating resistance to oxidative stress and cell death in Arabidopsis and Nicotiana benthamiana. Plant Mol Biol 63:273–287

    Article  PubMed  CAS  Google Scholar 

  • Nakane E, Kawakita K, Doke N, Yoshioka H (2003) Elicitation of primary and secondary metabolism during defense in the potato. J Gen Plant Pathol 69:378–384

    Article  CAS  Google Scholar 

  • Nasir KHB, Takahashi Y, Ito A, Saitoh H, Matsumura H, Kanzaki H, Shimizu T, Ito M, Fujisawa S, Sharma P, Ohme-Takagi M, Kamoun S, Terauchi R (2005) High-throughput in planta expression screening identifies a class II ethylene-responsive element binding factor-like protein that regulates plant cell death and non-host resistance. Plant J 43:491–505

    Article  PubMed  CAS  Google Scholar 

  • Ouelhadj A, Kuschk P, Humbeck K (2006) Heavy metal stress and leaf senescence induce the barley gene HvC2d1 encoding a calcium-dependent novel C2 domain-like protein. New Phytol 170:261–273

    Article  PubMed  CAS  Google Scholar 

  • Pontier D, Gan S, Amasino RM, Roby D, Lam E (1999) Markers for hypersensitive response and senescence show distinct patterns of expression. Plant Mol Biol 39:1243–1255

    Article  PubMed  CAS  Google Scholar 

  • Rea PA (2007) Plant ATP-binding cassette transporters. Annu Rev Plant Biol 58:347–375

    Article  PubMed  CAS  Google Scholar 

  • Rylott EL, Eastmond PJ, Gilday AD, Slocombe SP, Larson TR, Baker A, Graham IA (2006) The Arabidopsis thaliana multifunctional protein gene (MFP2) of peroxisomal β-oxidation is essential for seedling establishment. Plant J 45:930–941

    Article  PubMed  CAS  Google Scholar 

  • Sacco MA, Mansoor S, Moffett P (2007) A RanGAP protein physically interacts with the NB-LRR protein Rx, and is required for Rx-mediated viral resistance. Plant J 52:82–93

    Article  PubMed  CAS  Google Scholar 

  • Sano T, Kuraya Y, Amino S, Nagata T (1999) Phosphate as a limiting factor for the cell division of tobacco BY-2 cells. Plant Cell Physiol 40:1–8

    PubMed  Google Scholar 

  • Seo S, Okamoto M, Seto H, Ishizuka K, Sano H, Ohashi Y (1995) Tobacco MAP kinase: a possible mediator in wound signal transduction pathways. Science 270:1988–1992

    Article  PubMed  CAS  Google Scholar 

  • Seo S, Okamoto M, Iwai T, Iwano M, Fukui K, Isogai A, Nakajima N, Ohashi Y (2000) Reduced levels of chloroplast FtsH protein in tobacco mosaic virus-infected tobacco leaves accelerate the hypersensitive reaction. Plant cell 12:917–932

    Article  PubMed  CAS  Google Scholar 

  • Shah J (2005) Lipids, lipases, and lipid-modifying enzymes in plant disease resistance. Annu Rev Phytopathol 43:229–260

    Article  PubMed  CAS  Google Scholar 

  • Takahashi A, Casais C, Ichimura K, Shirasu K (2003) HSP90 interacts with RAR1 and SGT1 and is essential for RPS2-mediated disease resistance in Arabidopsis. Proc Natl Acad Sci USA 100:11777–11782

    Article  PubMed  CAS  Google Scholar 

  • Tameling WIL, Baulcombe DC (2007) Physical association of the NB-LRR resistance protein Rx with a Ran GTPase-activating protein is required for extreme resistance to Potato virus X. Plant Cell 19:1682–1694

    Article  PubMed  CAS  Google Scholar 

  • Tsuda S, Kirita M, Watanabe Y (1998) Characterization of a pepper mild mottle tobamovirus strain capable of overcoming the L 3 gene-mediated resistance, distinct from the resistance-breaking Italian isolate. Mol Plant Microbe Interact 11:327–331

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Xu Y, Han Y, Bao S, Du J, Yuan M, Xu Z, Chong K (2006) Overexpression of RAN1 in rice and Arabidopsis alters primordial meristem, mitotic progress, and sensitivity to auxin. Plant Physiol 140:91–101

    Article  PubMed  CAS  Google Scholar 

  • Weststeijn EA (1981) Lesion growth and virus localization in leaves of Nicotiana tabacum cv. Xanthi nc. after inoculation with Tobacco mosaic virus and incubation alternately at 22°C and 32°C. Physiol Plant Pathol 18:357–368

    Google Scholar 

  • Xia Y, Suzuki H, Borevitz J, Blount J, Guo Z, Patel K, Dixon RA, Lamb C (2004) An extracellular aspartic protease functions in Arabidopsis disease resistance signaling. EMBO J 23:980–988

    Article  PubMed  CAS  Google Scholar 

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

    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 

Download references

Acknowledgments

We thank Dr. K. W. Kinzler for SAGE2000 software, Harumi Takahashi and Kazue Obara for technical assistance and all members of IBRC for fruitful discussion. This study was supported in part by the Iwate Prefecture Government and in part by a grant-in-aid for Scientific Research (C) (18580047) from the Japan Society for the Promotion of Science.

Author information

Author notes

  1. Hiroyuki Hamada & Kazumi Suzuki

    Present address: National Agricultural Research Center, National Agriculture and Food Research Organization, 3-1-1 Kannondai, Tsukuba, Ibaraki, 305-8517, Japan

Authors and Affiliations

  1. Iwate Biotechnology Research Center, Kitakami, Iwate, 024-0003, Japan

    Hiroyuki Hamada, Hideo Matsumura, Reiko Tomita, Ryohei Terauchi, Kazumi Suzuki & Kappei Kobayashi

  2. School of Environmental Science, University of Shiga Prefecture, 2500 Hassaka-cho, Hikone, Shiga, 522-8533, Japan

    Kazumi Suzuki

Authors
  1. Hiroyuki Hamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hideo Matsumura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Reiko Tomita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ryohei Terauchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kazumi Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kappei Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kappei Kobayashi.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Tables S1 and S2 (DOC 47 kb)

10327_2008_106_MOESM2_ESM.tif

Fig. S1. Real-time PCR analysis for mRNA levels of SuperSAGE-tagged genes in normal infection process at 96 h postinoculation. C. chinense plants harboring the L 3 resistance gene were inoculated with P1,2 (filled column) or P1,2,3 (gray column) pathotypes of PMMoV or phosphate buffer (mock inoculation; open column). Total RNA was extracted from inoculated leaves at 96 h postinoculation for real-time PCR using primers in Table S1 and normalized against 18S rRNA levels. The mRNA levels relative to untreated plants were determined in two independent RNA preparations, and mean values are shown. (TIFF 240 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hamada, H., Matsumura, H., Tomita, R. et al. SuperSAGE revealed different classes of early resistance response genes in Capsicum chinense plants harboring L 3-resistance gene infected with Pepper mild mottle virus . J Gen Plant Pathol 74, 313–321 (2008). https://doi.org/10.1007/s10327-008-0106-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10327-008-0106-4

Keywords

Search

Navigation