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Phytoparasitica

, Volume 39, Issue 3, pp 269–283 | Cite as

Biochemical and molecular studies of early blight disease in tomato

  • Suchita J. PatelEmail author
  • R. B. Subramanian
  • Yachana S. Jha
Article

Abstract

Tomato early blight occurs worldwide and it is prevalent wherever tomatoes are grown. Alternaria solani Sorauer, the causal agent, has been recognized as a serious foliar pathogen of tomato and there are very few cultivars which possess resistance against early blight. Alternaric acid is the major toxin of A. solani. In this study, alternaric acid and fungal culture filtrate were used as an elicitor in NDT-96 (tolerant) and GP-5 (susceptible) tomato varieties in order to study and compare their abilities to induce defense-related enzymes, viz., catalase, peroxidase, β-1,3 glucanase, phenylalanine-ammonia-lyase (PAL), chitinase and polyphenol-oxidase (PPO) along with total phenols, and total soluble proteins. NDT-96 showed a rapid induction of all these pathogenesis-related enzymes except catalase along with total phenols as compared to GP-5 with both the treatments. Differential expression of total soluble proteins revealed higher protein content in NDT-96 as compared with GP-5. A 49.48 kDa protein was observed to be absent in GP-5. In addition, 25 microsatellite markers (SSR) were screened for polymorphisms among the above mentioned two tomato varieties. Of these, SSR 286 revealed a significant polymorphic band of 108 bp in NDT-96.

Keywords

Alternaria solani Pathogenesis-related enzymes Simple sequence repeats Systemic acquired resistance 

Notes

Acknowledgments

We are very thankful to Dr. Subhash Patel, Anand Agricultural University, India, for providing the Alternaria solani isolate.

References

  1. Barber, J. M. (1980). Catalase and peroxidase in primary leaves during development and senescence. Journal of Plant Breeding, 97, 135–144.Google Scholar
  2. Barksdale, T. H. (1971). Field evaluation for tomato early blight resistance. Plant Disease Reporter, 55, 807–809.Google Scholar
  3. Bashan, Y., Okon, Y., & Henis, Y. (1985). Peroxidase, polyphenol oxidase and phenols in relation to resistance against Pseudomonas syringae pv tomato in tomato. Canadian Journal of Botany, 65, 366–372.CrossRefGoogle Scholar
  4. Bell, J. N., Dixon, R. A., Bailey, J. A., Rowell, P. M., & Lamb, C. J. (1984). Differential induction of chalcone synthase mRNA activity at the onset of phytoalexin accumulation in compatible and incompatible plantpathogen interactions. Proceedings of the National Academy of Sciences of the United States of America, 81, 3384–3388.PubMedCrossRefGoogle Scholar
  5. Bhatia, I. S., Uppal, D. S., & Bajaj, K. L. (1972). Study of phenolic contents of resistant and susceptible varieties of tomato (Lycopersicum esculentum) in relation to early blight disease. Indian Phytopathology, 25, 231–235.Google Scholar
  6. Brian, P. W., Elson, G. W., Hemming, H. G., & Wright, J. M. (1952). The phytotoxic properties of alternaric acid in relation to the etiology of plant diseases caused by Alternaria solani (Ell. & Mart.) Jones & Grout. Applied Biology, 39, 308–321.CrossRefGoogle Scholar
  7. Carrasco, A., Boudet, A. M., & Marigo, G. (1978). Enhanced resistance of tomato plants to Fusarium by controlled stimulation of their natural phenolic production. Physiology and Plant Pathology, 12, 225–232.CrossRefGoogle Scholar
  8. Chen, Z., Silva, H., & Klessig, D. F. (1993). Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science, 262, 1883–1886.PubMedCrossRefGoogle Scholar
  9. Constabel, C. P., Yip, L., Patton, J. J., & Christopher, M. E. (2000). Polyphenol oxidase from hybrid poplar, cloning and expression in response to wounding and herbivory. Plant Physiology, 124, 285–295.PubMedCrossRefGoogle Scholar
  10. Daugrois, J. H., Lafitte, C., Barthe, J. P., & Touze, A. (1991). Induction of β-1,3-glucanase and chitinase activity in compatible and incompatible interactions between Colletotrichum lindemuthianum and bean cultivars. Journal of Phytopathology, 130, 225–234.CrossRefGoogle Scholar
  11. Deborah, S. D., Palaniswami, A., Vidhyasekaran, P., & Velazhahan, R. (2001). Time course study of the induction of defense enzymes, phenolics and lignin in rice in response to infection by pathogen and non-pathogen. Journal of Plant Diseases and Protection, 108, 204–216.Google Scholar
  12. Devi, M. C., & Reddy, M. N. (2002). Phenolic acid metabolism of groundnut (Arachis hypogaea L.) plants inoculated with VAM fungus and Rhizobium. Plant Growth Regulators, 37, 151–156.CrossRefGoogle Scholar
  13. Dickerson, D. P., Pascholati, S. F., Hagerman, A. E., Butler, L. G., & Nicholson, R. L. (1984). Phenylalanine ammonia- lyase and hydroxycinnamate: CoA ligase in maize mesocotyls inoculated with Helminthosporium maydis or Helminthosporium carbonum. Physiological Plant Pathology, 25, 111–123.CrossRefGoogle Scholar
  14. Dixon, R. A., Harrison, M. J., & Lamb, C. J. (1994). Early events in the activation of plant defense responses. Annual Review of Phytopathology, 32, 479–501.CrossRefGoogle Scholar
  15. Doyle, J. J., & Doyle, J. L. (1990). A rapid total DNA preparation procedure for fresh plant tissue. Focus, 12, 13–15.Google Scholar
  16. Du, H., & Klessig, D. L. (1997). Systemic acquired resistance in catalase deficient tobacco plant. Plant Physiology, 10, 922–925.Google Scholar
  17. Fernandez, A., Solorzano, E., Peteira, B., & Fernandez, E. (1996). Peroxidase induction in tomato leaves with different degrees of susceptibility to Alternaria solani. Revista de Protección Vegetal, 11, 79–83.Google Scholar
  18. Flott, B. E., Moersehbacher, B. M., & Reisener, H. (1989). Peroxidase isozyme patterns of resistant and susceptible wheat leaves following stem rust infection. The New Phytologist, 111, 413–421.CrossRefGoogle Scholar
  19. Frindlender, M., Inbar, J., & Chet, I. (1993). Biological control of soil borne plant pathogens by a β-1,3-glucanase producing Pseudomonas cepacia. Soil Biology and Biochemistry, 25, 1211–1221.CrossRefGoogle Scholar
  20. Gaube, C., Dubourg, C., Pawelec, A., Chamont, S., Blancard, D., & Briard, M. (2004). Brûlures foliaires parasitaires de la carotte. Alternaria dauci sous surveillance. PHM Revue Horticulture, 454, 15–18.Google Scholar
  21. Hammond-Kosack, K. E., & Jones, J. D. G. (1996). Resistance gene-dependent plant defense responses. The Plant Cell, 8, 1773–1791.PubMedCrossRefGoogle Scholar
  22. He, C., Poysa, V., & Yu, K. (2003). Development and characterization of simple sequence repeat (SSR) markers and their use in determining relationships among Lycopersicon esculentum cultivars. Theoretical and Applied Genetics, 106, 363–373.PubMedGoogle Scholar
  23. Herriot, A. B., Haynes, F. L., Jr., & Shoemaker, P. B. (1986). The heritability of resistance to early blight in diploid potatoes (Solanum tuberosum subsp. phureja and stenotonum). American Potato Journal, 63, 229–232.CrossRefGoogle Scholar
  24. Jaskani, M. J., Kwon, S. W., Kim, D. H., & Abbas, H. (2006). Seed treatments and orientation affects germination and seedling emergence in tetraploid watermelon. Pakistan Journal of Botany, 38, 89–98.Google Scholar
  25. Johanson, A., & Thurston, H. D. (1990). The effect of cultivar maturity on the resistance of potato to early blight caused by Alternaria solani. American Potato Journal, 67, 615–623.CrossRefGoogle Scholar
  26. Jung, W. J., Jin, Y. L., Kim, K. Y., Park, R. D., & Kim, T. H. (2005). Changes in pathogenesis-related proteins in pepper plants with regard to biological control of Phytophthora blight with Paenibacillus illinoisensis. Biocontrol, 50, 165–178.CrossRefGoogle Scholar
  27. Keinath, A., DuBose, V. B., & Rathwell, P. J. (1996). Efficacy and economics of three fungicide application schedules for early blight control and yield of fresh-market tomato. Plant Disease, 80, 1277–1282.CrossRefGoogle Scholar
  28. Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.PubMedCrossRefGoogle Scholar
  29. Lawrence, C. B., Joosten, M. H. A. J., & Tuzun, S. (1996). Differential induction of pathogenesis related proteins in tomato by Alternaria solani and the association of a basic chitinase isozyme with resistance. Physiological and Molecular Plant Pathology, 43, 361–377.CrossRefGoogle Scholar
  30. Lawrence, C. B., Singh, N. P., Qiu, J., Gardner, R. G., & Tuzun, S. (2000). Constitutive hydrolytic enzymes are associated with polygenic resistance of tomato to Alternaria solani and may function as an elicitor release mechanism. Physiological and Molecular Plant Pathology, 57, 211–220.CrossRefGoogle Scholar
  31. Linthorst, H. J. M. (1991). Pathogenesis-related proteins of plants. Critical Reviews in Plant Sciences, 10, 123–150.CrossRefGoogle Scholar
  32. Locke, S. B. (1948). A method for measuring resistance to defoliation diseases in tomato and other Lycopersicon species. Phytopathology, 38, 937–942.Google Scholar
  33. Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with folin phenol reagent. The Journal of Biological Chemistry, 193, 265–275.PubMedGoogle Scholar
  34. Madden, L., Pennypacker, S. P., & MacNab, A. A. (1978). FAST, a forecast system for Alternaria solani on tomato. Phytopathology, 68, 1354–1358.CrossRefGoogle Scholar
  35. Maher, E. A., Bate, N. J., Ni, W., Elkind, Y., Dixon, R. A., & Lamb, C. J. (1994). Increased disease susceptibility of transgenic tobacco plants with suppressed levels of preformed phenylpropanoid products. Proceedings of the National Academy of Sciences of the United States of America, 91, 7802–7806.PubMedCrossRefGoogle Scholar
  36. Mauch-Mani, B., & Slusarenko, A. J. (1996). Production of salicylic acid precursors is a major function of phenylalanine ammonia lyase in the resistance of Arabidopsis to Perenospora parasitica. The Plant Cell, 8, 203–212.PubMedCrossRefGoogle Scholar
  37. Mayer, A. M., Harel, E., & Shaul, R. B. (1965). Assay of catechol oxidase—a critical comparison of methods. Phytochemistry, 5, 783–789.CrossRefGoogle Scholar
  38. Nash, A. F., & Gardner, R. G. (1988). Tomato early blight resistance in a breeding line derived from Lycopersicon hirsutum PI 126445. Plant Disease, 72, 206–209.CrossRefGoogle Scholar
  39. Neuhoff, V., Arold, N., Taube, D., & Ehrhardt, W. (1988). Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis, 9, 255–262.PubMedCrossRefGoogle Scholar
  40. Pan, S. Q., Ye, X. S., & Kuç, J. (1991). Association of β-1,3 glucanase activity and isoform pattern with systemic resistance to blue mold in tobacco induced by stem injection with Peronospora tabacina or leaf inoculation with tobacco mosaic virus. Physiological and Molecular Plant Pathology, 39, 25–39.CrossRefGoogle Scholar
  41. Pound, G. S., & Stahmann, M. A. (1951). The production of toxic material by Alternaria solani and its relation to the early blight disease of tomato. Phytopathology, 41, 1104–1114.Google Scholar
  42. Powell, W., Machray, G. C., & Provan, J. (1996). Polymorphism revealed by simple sequence repeats. Trends in Plant Science, 1, 215–222.Google Scholar
  43. Powell, W., Orozco-Castillo, C., Chalmers, K. J., Provan, J., & Waugh, R. (1995). Polymerase chain reaction-based assays for the characterisation of plant genetic resources. Electrophoresis, 16, 1726–1730.PubMedCrossRefGoogle Scholar
  44. Radhajeyalakshmi, R., Velazhahan, R., Samiyappan, R., & Doraiswamy, S. (2009). Systemic induction of pathogenesis related proteins (PRs) in Alternaria solani elicitor sensitized tomato cells as resistance response. Scientific Research and Essay, 4, 685–689.Google Scholar
  45. Rafalski, J. A., Vogel, J. M., Morgante, M., Powell, W., Andre, C., & Tingey, S. V. (1996). Generating new DNA markers in plants. In B. Birren & E. Lai (Eds.), Non-mammalian genomic analysis: A practical guide (pp. 75–134). New York, NY: Academic.Google Scholar
  46. Rajput, S. G., Wable, K. J., Sharma, K. M., Kubde, P. D., & Mulay, S. A. (2006). Reproducibility testing of RAPD and SSR markers in tomato. African Journal of Biotechnology, 5, 108–112.Google Scholar
  47. Rani, P., & Yasur, J. (2009). Physiological changes in groundnut plants induced by pathogenic infection of Cercosporidium personatum Deighton. Allelopathy Journal, 23, 369–378.Google Scholar
  48. Reimers, P. J., Guo, A., & Leach, J. E. (1992). Increased activity of cationic peroxidase associated with an incompatible interaction between Xanthomonas oryzae pv oryzae and rice (Oryza sativa). Plant Physiology, 99, 1044–1050.PubMedCrossRefGoogle Scholar
  49. Reissig, J. L., Strominger, J. L., & Leloir, L. F. (1959). A modified colorimetric method for the estimation of N-acetyl sugars. The Journal of Biological Chemistry, 217, 959–962.Google Scholar
  50. Ryals, J. A., Neuenschwander, U. H., Willits, M. G., Molina, A., Steiner, H. Y., & Hunt, M. D. (1996). Systemic acquired resistance. The Plant Cell, 8, 1809–1819.PubMedCrossRefGoogle Scholar
  51. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Hori, G. T., et al. (1988). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science, 239, 487–491.PubMedCrossRefGoogle Scholar
  52. Sherf, A. F., & MacNab, A. A. (1986). Vegetable diseases and their control. New York, NY: Wiley.Google Scholar
  53. Shtienberg, D., Blachinsky, D., Kremer, Y., Ben-Hador, G., & Dinoor, A. (1995). Integration of genotype and age-related resistance to reduce fungicide use in management of Alternaria diseases of cotton and potato. Phytopathology, 85, 995–1002.CrossRefGoogle Scholar
  54. Siriphanich, J., & Kader, A. A. (1985). Effects of CO2 on cinnamic acid 4-hydroxylase in relation to phenolic metabolism in lettuce tissue. Journal of the American Society for Horticultural Science, 110, 333–335.Google Scholar
  55. Solorzano, E., Fernandez, A., Peteira, B., & Fernandez, E. (1996). Polyphenol oxidases and phenylalanine ammonium lyases induction in tomato leaves infected with Alternaria solani. Revista de Protección Vegetal, 11, 153–157.Google Scholar
  56. Stewart, R. J., Sawyer, B. J. B., Bucheli, C. S., & Robinson, S. P. (2001). Polyphenol oxidase is induced by chilling and wounding in pineapple. Australian Journal of Plant Physiology, 28, 181–191.Google Scholar
  57. Stout, M. J., Fidantsef, A. L., Duffey, S. S., & Bostock, R. M. (1999). Signal interactions in pathogen and insect attack: systemic plant-mediated interactions between pathogens and herbivores of the tomato, Lycopersicon esculentum. Physiological and Molecular Plant Pathology, 54, 115–130.CrossRefGoogle Scholar
  58. Sumner, J. B., & Gjessing, E. C. (1943). A method for the determination of peroxidase activity. Archives of Biochemistry and Biophysics, 2, 291–293.Google Scholar
  59. Thipyapong, P., Hunt, M. D., & Steffens, J. C. (1995). Systemic wound induction of potato (Solanum tuberosum) polyphenol oxidase. Phytochemistry, 40, 673–676.CrossRefGoogle Scholar
  60. Thipyapong, P., & Steffens, J. C. (1997). Tomato polyphenol oxidase: differential response of the polyphenol oxidase F promoter to injuries and wound signals. Plant Physiology, 115, 409–418.PubMedGoogle Scholar
  61. Vanacker, H., Carver, T. L. W., & Foyer, C. H. (1998). Pathogen-induced changes in the antioxidant status of the apoplast in barley leaves. Plant Physiology, 117, 1103–1114.PubMedCrossRefGoogle Scholar
  62. Van Loon, L. C. (1997). Induced resistance in plants and the role of pathogenesis related proteins. European Journal of Plant Pathology, 103, 753–765.CrossRefGoogle Scholar
  63. Velazhahan, R., & Vidhyasekaran, P. (1994). Role of phenolic compounds, peroxidase and polyphenol oxidase in resistance of groundnut to rust. Acta Phytopathologica et Entomologica Hungarica, 29, 23–29.Google Scholar
  64. Wang, Y., Tang, X., Cheng, Z., Mueller, L., Giovannoni, J., & Tanksley, S. D. (2006). Euchromatin and pericentromeric heterochromatin: comparative composition in the tomato genome. Genetics, 172, 2529–2540.PubMedCrossRefGoogle Scholar
  65. Ward, E., Uknes, S. J., Williams, S. C., Dincher, S. S., Wiederhold, D. L., Alexander, D. C., et al. (1991). Coordinate gene activity in response to agents that induce systemic acquired resistance. The Plant Cell, 3, 1085–1094.PubMedCrossRefGoogle Scholar
  66. Yao, K., De Luca, V., & Brisson, N. (1995). Creation of a metabolic sink for tryptophan alters the phenyl propanoid pathway and the susceptibility of potato to Phytophthora infestans. The Plant Cell, 7, 1787–1799.PubMedCrossRefGoogle Scholar
  67. Zieslin, N., & Ben-Zaken, R. (1993). Peroxidase activity and presence of phenolic substances in peduncles of rose flowers. Plant Physiology and Biochemistry, 31, 333–339.Google Scholar

Copyright information

© Springer Science & Business Media BV 2011

Authors and Affiliations

  • Suchita J. Patel
    • 1
    Email author
  • R. B. Subramanian
    • 1
  • Yachana S. Jha
    • 1
  1. 1.B. R. Doshi School of Biosciences, Satellite CampusSardar Patel UniversityVallabh VidhyanagarIndia

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