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Plant Molecular Biology

, Volume 62, Issue 4–5, pp 669–682 | Cite as

Single amino acid alterations in Arabidopsis thaliana RCY1 compromise resistance to Cucumber mosaic virus, but differentially suppress hypersensitive response-like cell death

  • Ken-Taro Sekine
  • Takeaki Ishihara
  • Shu Hase
  • Tomonobu Kusano
  • Jyoti Shah
  • Hideki TakahashiEmail author
Article

Abstract

Resistance to an yellow strain of Cucumber mosaic virus [CMV(Y)] in Arabidopsis thaliana ecotype C24 is conferred by the CC-NBS-LRR type R gene, RCY1. RCY1-conferred resistance is accompanied by a hypersensitive response (HR), which is characterized by the development of necrotic local lesion (NLL) at the site of infection that restricts viral spread. To further characterize the role of RCY1 in NLL formation we have identified six recessive CMV(Y)-susceptible rcy1 mutants, four of which contain single amino acid substitutions in RCY1: rcy1-2 contains a D to N substitution in the CC domain, rcy1-3 and rcy1-4 contain R to K and E to K substitutions, respectively, in the LRR domain, and rcy1-6 contains a W to C substitution in the NBS domain. The rcy1-5 and rcy1-7 contain nonsense mutations in the LRR and NBS domains, respectively. Although the virus systemically spread in all six rcy1 mutants, HR-associated cell death was differentially induced in these mutants. In comparison to the wild type C24 plant, HR was not observed in the CMV(Y)-inoculated leaves of the rcy1-3, rcy1-5, rcy1-6 and rcy1-7 mutants. In contrast, delayed NLL development was observed in the virus inoculated leaves of the rcy1-2 and rcy1-4 mutants. In addition, necrosis accompanied by elevated accumulation of PR gene transcript also appeared in the non-inoculated leaves of the rcy1-2 and rcy1-4 mutants. Trans-complementation was observed between the rcy1-2 and rcy1-4 alleles; in F1 plants derived from a cross between rcy1-2 and rcy1−4, HR associated cell death was accelerated and systemic spread of the virus was partially suppressed than in the homozygous rcy1-2 and rcy1-4 plants. Our results suggest that the CC, NBS and LRR domains of RCY1 are required for restriction of virus spread but differentially impact the induction of HR-like cell death. Furthermore, these results also predict inter-molecular interaction involving RCY1 in Arabidopsis resistance to CMV(Y).

Keywords

Resistance gene Gene-for-gene interaction Viral resistance 

Abbreviations

CC

Coiled-coil

CMV(Y)

An yellow strain of Cucumber mosaic virus

EMS

Ethyl methanesulfonate

NBS

Nucleotide-binding site

NLL

Necrotic local lesion

LRR

Leucine-rich repeats

RCY1

RESISTANCE TO CMV(Y) 1

TIR

Toll-interleukin-1 receptor

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Notes

Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas (Molecular Mechanisms of Plant–Pathogenic Microbe Interaction – Toward Production of Disease Resistant Plants) and for Scientific Research (B) (16380031) from Ministry of Education, Culture, Sports and Arts, Japan.

References

  1. An G (1986) Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cell. Plant Physiol 81:86–91PubMedCrossRefGoogle Scholar
  2. Aravind L, Dixit VM, Koonin EV (1999) The domains of death: evolution of the apoptosis machinery. Trends Biochem. Sci 24:47–53PubMedCrossRefGoogle Scholar
  3. Axtell MJ, McNellis TW, Mudgett MB, Hsu CS, Staskawicz BJ (2001) Mutational analysis of the Arabidopsis RPS2 disease resistance gene and the corresponding Pseudomonas syringae avrRpt2 avirulence gene. Mol. Plant-Microbe Interact 14:181–188PubMedGoogle Scholar
  4. Axtell MJ, Staskawicz BJ (2003) Initiation of RPS2-specified disease resistance in Arabidopsis is coupled to AvrRpt2-directed elimination of RIN4. Cell 112:369–377PubMedCrossRefGoogle Scholar
  5. Banerjee D, Zhang XC, Bent AF (2001) The leucine-rich repeat domain can determine effective interaction between RPS2 and other host factors in Arabidopsis RPS2-mediated disease resistance. Genetics 158:439–450PubMedGoogle Scholar
  6. Bechtold N (1998) In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. In: Martinez-Zapater J, Salinas J. (eds) Methods in Molecular Biology. Humana Press, Totowa NJ, pp. 259–266Google Scholar
  7. Belkhadir Y, Subramaniam R, Dangl JL (2004) Plant disease resistance protein signaling: NBS-LRR proteins and their partners. Curr. Opin. Plant Biol 7:391–399PubMedCrossRefGoogle Scholar
  8. Bendahmane A, Kanyuka K, Baulcombe DC (1999) The Rx gene from potato controls separate virus resistance and cell death responses. Plant Cell 11:781–791PubMedCrossRefGoogle Scholar
  9. Bendahmane A, Farnham G, Moffett P, Baulcombe DC (2002) Constitutive gain-of-function mutants in a nucleotide binding site-leucine rich repeat protein encoded at the Rx locus of potato. Plant J 32:195–204PubMedCrossRefGoogle Scholar
  10. Brommonschenkel SH, Frary A, Frary A, Tanksley SD (2000) The broad-spectrum tospovirus resistance gene Sw-5 of tomato is a homolog of the root-knot nematode resistance gene Mi. Mol. Plant-Microbe Interact 13:1130–1138PubMedGoogle Scholar
  11. Cooley MB, Pathirana S, Wu H-J, Kachroo P, Klessig DF (2000) Members of the Arabidopsis HRT/RPP8 family of resistance genes confer resistance to both viral and oomycete pathogens. Plant Cell 12:663–676PubMedCrossRefGoogle Scholar
  12. Dangl JL, Jones JDG (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826–833PubMedCrossRefGoogle Scholar
  13. Dinesh-Kumar SP, Tham W-H, Baker BJ (2000) Structure-function analysis of the tobacco mosaic virus resistance gene N. Proc. Natl. Acad. Sci. USA 97:14789–14794PubMedCrossRefGoogle Scholar
  14. Dodds PN, Lawrence GJ, Ellis JG (2001) Six amino acid changes confined to the leucine-rich repeat β-strand/β-turn motif determine the difference between the P and P2 rust resistance specificities in flax. Plant Cell 13:163–178PubMedCrossRefGoogle Scholar
  15. Ellis JG, Lawrence GJ, Luck JE, Dodds PN (1999) Identification of regions in alleles of the flax rust resistance gene L that determine differences in gene-for-gene specificity. Plant Cell 11:495–506PubMedCrossRefGoogle Scholar
  16. Ellis JG, Dodds PN, Pryor T (2000) Structure, function, and evolution of plant disease resistance genes. Curr. Opin. Plant Biol 3:278–284CrossRefGoogle Scholar
  17. Hammond-Kosack KE, Jones JDG (1997) Plant disease resistance genes. Annu. Rev. Plant Physiol. Plant Mol. Biol 48:575–607PubMedCrossRefGoogle Scholar
  18. Hammond-Kosack KE, Parker JE (2003) Deciphering plant-pathogen communication: fresh perspectives for molecular resistance breeding. Curr. Opin. Biotech 14:177–193PubMedCrossRefGoogle Scholar
  19. Hwang C-F, Bhakta AV, Truesdell GM, Pudlo WM, Williamson VM (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 12:1319–1329PubMedCrossRefGoogle Scholar
  20. Inohara N, Chamaillard M, McDonald C, Nunez G (2005) NOD-LRR proteins: role in host-microbial interactions and inflammatory disease. Annu. Rev. Biochem 74:355–383PubMedCrossRefGoogle Scholar
  21. Ishihara T, Sakurai N, Sekine K-T, Hase S, Ikegami M, Shibata D, Takahashi H (2004) Comparative analysis of expressed sequence tags in resistant and susceptible ecotypes of Arabidopsis thaliana infected with cucumber mosaic virus. Plant Cell Physiol 45:470–480PubMedCrossRefGoogle Scholar
  22. Lanfermeijer FC, Dijkhuis J, Sturre MJG, de Haan P, Hille J (2003) Cloning and characterization of the durable tomato mosaic virus resistance gene Tm-2 2 from Lycopersicon esculentum. Plant Mol. Biol 52:1037–1049PubMedCrossRefGoogle Scholar
  23. Luck JE, Lawrence GJ, Dodds PN, Shepherd KW, Ellis JG (2000) Regions outside of the leucine-rich repeats of flax rust resistance proteins play a role in specificity determination. Plant Cell 12:1367–1377PubMedCrossRefGoogle Scholar
  24. Mackey D, Belkhadir Y, Alonso JM, Ecker JR, Dangl JL (2003) Arabidopsis RIN4 is a target of the type III virulence effector AvrRpt2 and modulates RPS2-mediated resistance. Cell 112:379–389PubMedCrossRefGoogle Scholar
  25. Matthews REF. 2002. Plant Virology, 4th edn, Academic Press, New York. Google Scholar
  26. McDowell JM, Dhandaydham M, Long TA, Aarts MGM, Goff S, Holub EB, Dangl JL (1998) Intragenic recombination and diversifying selection contribute to the evolution of downy mildew resistance at the RPP8 locus of Arabidopsis. Plant Cell 10:1861–1874PubMedCrossRefGoogle Scholar
  27. Mestre P, Baulcombe DC (2006) Elicitor-mediated oligomerization of the tobacco N disease resistance protein. Plant Cell 18:491–501PubMedCrossRefGoogle Scholar
  28. Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant Cell 15:809–834PubMedCrossRefGoogle Scholar
  29. Moffett P, Farnham G, Peart J, Baulcombe DC (2002) Interaction between domains of a plant NBS-LRR protein in disease resistance-related cell death. EMBO J 21:4511–4519PubMedCrossRefGoogle Scholar
  30. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plantrum 15:473–497CrossRefGoogle Scholar
  31. Murray MG, Thompson WF (1980) Rapid isolation of high␣molecular weight plant DNA. Nucl. Acids Res 8:4321–4325PubMedGoogle Scholar
  32. Noel L, Moores TL, van der Biezen EA, Parniske M, Daniels MJ, Parker JE, Jones JDG (1999) Pronounced intraspecific haplotype divergence at the RPP5 complex disease resistance locus of Arabidopsis. Plant Cell 11:2099–2111PubMedCrossRefGoogle Scholar
  33. Palukaitis P, García-Arenal F (2003) Cucumoviruses. Adv. Virus Res 62:241–323PubMedCrossRefGoogle Scholar
  34. Rédei GP, Koncz C (1992) Classical Mutagenesis. In: Koncz C, Chua N-H, Shell J (eds) Methods in Arabidopsis Research. World Scientific, Singapore, pp 16–82Google Scholar
  35. Schulze-Lefert P (2004) Plant immunity: the origami of receptor activation. Curr. Biol 14:22–24CrossRefGoogle Scholar
  36. Sekine K-T, Nandi A, Ishihara T, Hase S, Ikegami M, Shah J, Takahashi H (2004) Enhanced resistance to Cucumber mosaic virus in the Arabidopsis thaliana ssi2 mutant is mediated via an SA-independent mechanism. Mol. Plant-Microbe Interact 17:623–632Google Scholar
  37. Takahashi H, Ehara Y (1993) Severe chlorotic spot symptoms in cucumber mosaic virus strain Y-infected tobaccos are induced by a combination of the virus coat protein gene and two host recessive genes. Mol. Plant-Microbe Interact 6:182–189PubMedGoogle Scholar
  38. Takahashi H, Goto N, Ehara Y (1994) Hypersensitive response in cucumber mosaic virus-inoculated Arabidopsis thaliana. Plant J 6:369–377CrossRefGoogle Scholar
  39. Takahashi H, Kanayama Y, Zheng MS, Kusano T, Hase S, Ikegami M, Shah J (2004a) Antagonistic interactions between the SA and JA signaling pathways in Arabidopsis modulate expression of defense genes and gene-for-gene resistance to Cucumber mosaic virus. Plant Cell Physiol 45:803–809CrossRefGoogle Scholar
  40. Takahashi H, Miller J, Nozaki Y, Sukamto, Takeda M, Shah J, Hase S, Ikegami M, Ehara Y, Dinesh-Kumar SP (2002) RCY1, an Arabidopsis thaliana RPP8/HRT family resistance gene, conferring resistance to cucumber mosaic virus requires salicylic acid, ethylene and a novel signal transduction mechanism. Plant J 32:655–667PubMedCrossRefGoogle Scholar
  41. Takahashi H, Sekine K-T, Ishihara T, Hase S, Ikegami M, Shah J (2004b) Signal transduction pathways governing resistance to Cucumber mosaic virus. In: Tsuyumu S, Leach JE, Shiraishi T, Wolpert T (eds) Genomics and Genetic Analysis of Plant Parasitism and Defense. APS Press, Saint Paul pp.185–194Google Scholar
  42. Takahashi H, Suzuki M, Natsuaki K, Shigyo T, Hino K, Teraoka T, Hosokawa D, Ehara Y (2001) Mapping the virus and host genes involved in the resistance response in Cucumber mosaic virus-infected Arabidopsis thaliana. Plant Cell Physiol 42:340–347PubMedCrossRefGoogle Scholar
  43. Tanabe T, Chamaillard M, Ogura Y, Zhu L Qiu S, Masumoto J, Ghosh P, Moran A, Predergast MM, Tromp G, Williams CJ, Inohara N, Nunez G (2004) Regulatory regions and critical residues of NOD2 involved in muramyl dipeptide recognition. EMBO J 23:1587–1597PubMedCrossRefGoogle Scholar
  44. Tao Y, Yuan F, Leister RT, Ausubel FM, Katagiri F (2000) Mutational analysis of the Arabidopsis nucleotide binding site-leucine-rich repeat resistance gene RPS2. Plant Cell 12:2541–2554PubMedCrossRefGoogle Scholar
  45. Tameling WIL, Elzinga SDJ, Darmin PS, Vossen JH, Takken FLW, Haring MA, Cornelissen BJC (2002) The tomato R gene products I-2 and Mi-1 are functional ATP binding proteins with ATPase activity. Plant Cell 14:2929–2939PubMedCrossRefGoogle Scholar
  46. Tomaru K, Hidaka J (1960) Strains of cucumber mosaic virus isolated from tobacco plants. III. A yellow strain. Bull. Hatano Tobacco Exp. Station 46:143–149Google Scholar
  47. Tornero P, Chao RA, Luthin WN, Goff SA, Dangl JL (2002) Large-scale structure-function analysis of the Arabidopsis RPM1 disease resistance protein. Plant Cell 14:435–450PubMedCrossRefGoogle Scholar
  48. Traut TW (1994) The functions and consensus motifs of nine types of peptide segments that form different types of nucleotide-binding sites. Eur. J. Biochem 222:9–19PubMedCrossRefGoogle Scholar
  49. Van der Biezen EA, Jones JDG (1998) The NB-ARC domain: a novel signaling motif shared by plant resistance gene products and regulators of cell death in animals. Curr. Biol 8:R226–227PubMedCrossRefGoogle Scholar
  50. Warren RF, Henk A, Mowery P, Holub E, Innes RW (1998) A mutation within the leucine-rich repeat domain of the Arabidopsis disease resistance gene RPS5 partially suppresses multiple bacterial and downy mildew resistance genes. Plant Cell 10:1439–1452PubMedCrossRefGoogle Scholar
  51. Whitham S, Dinesh-kumar SP, Choi D, Hehl R, Corr C, Baker B (1994) The product of the tobacco mosaic virus resistance gene N: Similarity to toll and the interleukin-1 receptor. Cell 78:1101–1115PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Ken-Taro Sekine
    • 1
  • Takeaki Ishihara
    • 1
  • Shu Hase
    • 1
  • Tomonobu Kusano
    • 2
  • Jyoti Shah
    • 3
  • Hideki Takahashi
    • 1
    Email author
  1. 1.Department of Life Science, Graduate School of Agricultural ScienceTohoku UniversityMiyagiJapan
  2. 2.Graduate School of Life SciencesTohoku UniversityMiyagi Japan
  3. 3.Division of Biology and the Molecular, Cellular and Developmental Biology Program, Kansas State UniversityManhattanU.S.A

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