Skip to main content
Virologica Sinica logoLink to Virologica Sinica
. 2014 Aug 4;29(4):250–256. doi: 10.1007/s12250-014-3452-8

Effect of chitosan on tobacco mosaic virus (TMV) accumulation, hydrolase activity, and morphological abnormalities of the viral particles in leaves of N. tabacum L. cv. Samsun

Vera Nagorskaya 1, Anatoliy Reunov 1,, Larisa Lapshina 1, Viktoriya Davydova 1, Irina Yermak 1
PMCID: PMC8206317  PMID: 25116808

Abstract

The effect of chitosan on the development of infection caused by Tobacco mosaic virus (TMV) in leaves of Nicotiana tabacum L. cv. Samsun has been studied. It was shown that the infectivity and viral coat protein content in leaves inoculated with a mixture of TMV (2 μg/mL) and chitosan (1 mg/mL) were lower in the early period of infection (3 days after inoculation), by 63% and 66% respectively, than in leaves inoculated with TMV only. Treatment of leaves with chitosan 24 h before inoculation with TMV also caused the antiviral effects, but these were less apparent than when the virus and polysaccharide were applied simultaneously. The inhibitory effects of the agent decreased as the infection progressed. Inoculation of leaves with TMV together with chitosan considerably enhanced the activity of hydrolases (proteases, RNases) in the leaves, in comparison with leaves inoculated with TMV alone. Electron microscope assays of phosphotungstic acid (PTA)-stained suspensions from infected tobacco leaves showed that, in addition to the normal TMV particles (18 nm in diameter, 300 nm long), these suspensions contained abnormal (swollen, “thin” and “short”) virions. The highest number of abnormal virions was found in suspensions from leaves inoculated with a mixture of TMV and chitosan. Immuno-electron microscopy showed that “thin” virus particles, in contrast to the particles of normal diameter, lost the ability to bind to specific antiserum. It seems that the chitosan-induced activation of hydrolases stimulates the intracellular degradation of TMV particles and hence hydrolase activation may be considered to be one of the polysaccharide-mediated cellular defense mechanisms that limit virus accumulation in cells.

Keywords: tobacco mosaic virus (TMV), chitosan, antiviral activity, infection

References

  1. Begam M, Narwal S, Roy S, Kumar S, Lodha M L, Kapoor H C. An antiviral protein having deoxyribonuclease and ribonuclease activity from leaves of the post-flowering stage of Celosia cristata. Biochemistry (Moscow) 2006;71:44–48. doi: 10.1134/S0006297906130074. [DOI] [PubMed] [Google Scholar]
  2. Choudhary N L, Yadav O P, Lodha M L. Ribonuclease, deoxyribonuclease, and antiviral activity of Escherichia coli expressed Bougainvillea xbuttiana antiviral protein. Biochemistry (Moscow) 2008;73:273–277. doi: 10.1134/S000629790803005X. [DOI] [PubMed] [Google Scholar]
  3. Davydova V N, Naberezhnykh G A, Yermak I M, Gorbach V I, Solov’eva T F. Comparative study of physicochemical properties of chitosans of varying degree of polymerization in neutral aqueous solutions. Biofizika. 2000;45:641–647. [PubMed] [Google Scholar]
  4. Davydova V N, Nagorskaya V P, Gorbach V I, Kalitnik A A, Reunov A V, Solov’eva T F, Yermak I M. Chitosan anti viral activity: Dependence on structure and depolymerisation method. Appl Biochem Microbiol. 2011;47:103–108. doi: 10.1134/S0003683811010042. [DOI] [PubMed] [Google Scholar]
  5. Edreva A. A novel strategy for plant protection: Induced resistance. J Cell Molec Biol. 2004;3:61–69. [Google Scholar]
  6. Esau K. Viruses in plant hosts. Form, distribution and pathologic effects. Madison, Milwaukee, and London: Univ. Wisconsin Press; 1968. p. 225. [Google Scholar]
  7. Everitt E, Persson M J, Wohlfart C. pH-dependent exposure of endoproteolytic cleavage sites of the adenovirus 2 hexon protein. FEMS Microbiol Lett. 1988;49:229–233. doi: 10.1111/j.1574-6968.1988.tb02721.x. [DOI] [Google Scholar]
  8. Golinowski W, Kupidlowska E, Skrzeczkowski L. Degradation of potato virus X (PVX) in the intercellular spaces of potato mesophyll. Phytopathol Z. 1981;101:136–142. doi: 10.1111/j.1439-0434.1981.tb03331.x. [DOI] [Google Scholar]
  9. Hadrami A E, Adam L R, Hadrami I E, Daayf F. Chitosan in plant protection. Mar Drugs. 2010;8:968–987. doi: 10.3390/md8040968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hatsugai N, Kuroyanagi M, Yamada K, Meshi T, Tsuda S, Kondo M, Nishimura M, Hara-Nishimura I. A plant vacuolar protease, VPE, mediated virus-induced hypersensitive cell death. Science. 2004;305:855–858. doi: 10.1126/science.1099859. [DOI] [PubMed] [Google Scholar]
  11. Iriti M, Sironib M, Gomarasca S, Casazza A P, Soave C, Faoro F. Cell death-mediated antiviral effect of chitosan in tobacco. Plant Physiol Biochem. 2006;44:893–900. doi: 10.1016/j.plaphy.2006.10.009. [DOI] [PubMed] [Google Scholar]
  12. Kulikov S N, Chirkov S N, Il’ina A V, Lopatin S A, Varlamov V P. Effect of the molecular weight of chitosan on its antiviral activity in plants. Appl Biochem Microbiol. 2006;42:200–203. doi: 10.1134/S0003683806020165. [DOI] [PubMed] [Google Scholar]
  13. Lapshina L A, Reunov A V, Nagorskaya V P, Zvyagintseva T N, Shevchenko N M. Inhibitory effect of fucoidan from alga Fucus evanescens on the spread of infection induced by tobacco mosaic virus in tobacco leaves of two cultivars. Russ. J Plant Physiol. 2006;53:246–251. doi: 10.1134/S1021443706020154. [DOI] [Google Scholar]
  14. Lapshina L A, Nagorskaya V P, Reunov A V, Barabanova A O, Shevchenko N M, Yermak I M, Zvyagintseva T N, Elyakova L A. Correlation between influence of polysaccharides on hydrolase activity and their antiviral effect in tobacco leaves. Biochemistry (Moscow) 2011;76:566–572. doi: 10.1134/s0006297911040092. [DOI] [PubMed] [Google Scholar]
  15. Mayhew D E, Ford R E. An inhibitor of tobacco mosaic virus produced by Physarum potycephalum. Phytopathology. 1971;61:636–640. doi: 10.1094/Phyto-61-636. [DOI] [Google Scholar]
  16. Otsuki Y, Takebe I, Onho T, Fukuda M, Okada Y. Reconstitution of tobacco mosaic rods occurs bidirectionally from an internal initiation region: demonstration by electron microscopic serology. Proc Natl Acad Sci USA. 1977;74:1913–1917. doi: 10.1073/pnas.74.5.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Park C J, Kim K J, Shin R, Park J M, Shin Y C, Paek K H. Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway. Plant J. 2004;37:186–198. doi: 10.1046/j.1365-313X.2003.01951.x. [DOI] [PubMed] [Google Scholar]
  18. Reunov A V, Lapshina L A, Nagorskaya V P, Elyakova L A. Effect of 1, 3;1,6-β-glucan on infection of detached tobacco leaves with tobacco mosaic virus. J Phytopathol. 1996;144:247–249. doi: 10.1111/j.1439-0434.1996.tb01524.x. [DOI] [Google Scholar]
  19. Reunov A V, Lapshina L A, Nagorskaya V P. Comparative study of accumulation of two Potato virus X strains differing in virulence, hydrolase activity and morphological abnormalities in Datura stramonium L. leaves. J Phytopathol. 2013;161:348–352. doi: 10.1111/jph.12073. [DOI] [Google Scholar]
  20. Reunov A V, Nagorskaya V P, Lapshina L A, Yermak I M, Barabanova A O. Effect of k/β-carragenan from red alga Tichocarpus crinitus (Tichocarpaceae) on infection of detached tobacco leaves with tobacco mosaic virus. J Plant Dis Protect. 2004;111:165–172. [Google Scholar]
  21. Reunov A V, Lapshina L A, Nagorskaya V P, Zvyagintseva T N, Shevchenko N M. Effect of fucoidan from the brown alga Fucus evanescens on the development of infection induced by potato virus X in Datura stramonium L. J Plant Dis Protect. 2009;116:49–54. [Google Scholar]
  22. Roggero P, Pennazio S. Quantitative determination by ELISA of tobacco necrosis virus from necrotic local lesions in tobacco. J Virol Methods. 1984;8:283–291. doi: 10.1016/0166-0934(84)90065-X. [DOI] [PubMed] [Google Scholar]
  23. Rouhier P, Kopp M, Begot V, Bruneteau M, Fritig B. Structural features of fungal β-D-glucans for the efficient inhibition of the initiation of virus infection on Nicotiana tabacum. Phytochemistry. 1995;39:57–62. doi: 10.1016/0031-9422(94)00852-K. [DOI] [PubMed] [Google Scholar]
  24. Salomon R. Partial cleavage of sweet potato feathery mottle virus coat protein subunit by an enzyme in extracts of infected symptomless leaves. J Gen Virol. 1989;70:1943–1949. doi: 10.1099/0022-1317-70-8-1943. [DOI] [PubMed] [Google Scholar]
  25. Slováková L, Lišková D, Capek P, Kubačková M, Kákoniová D, Karácsonyi Defence responses against TNV infection induced by galactoglucomannan-derived oligosaccharides in cucumber cells. Eur J Plant Pathol. 2000;106:543–553. doi: 10.1023/A:1008722318178. [DOI] [Google Scholar]
  26. Stübler D, Buchenauer H. Antiviral activity of the glucan lichenan (Poly-β{1→3, 1→4}D-Anhydroglucose) 1. Biological activity in tobacco plants. J Phytopathol. 1996;144:37–43. doi: 10.1111/j.1439-0434.1996.tb01486.x. [DOI] [Google Scholar]
  27. Šubíková V, Slováková L, Farkaš V. Inhibition of tobacco necrosis virus infection by xyloglucan fragments. Z PflKrankh Pflschurz. 1994;101:128–131. [Google Scholar]
  28. Thakur M, Sohal B S. ISRN Biochemistry. 2013. Role of elicitors in inducing resistance in plants against pathogen infection: A Review. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Trifonova E A, Sapotsky M V, Komarova M L, Scherban A V, Shumny V K, Polyakova A M, Lapshina L A, Kochetov A V, Malinovsky V I. Protection of transgenic tobacco plants expressing bovine pancreatic ribonuclease against tobacco mosaic virus. Plant Cell Rep. 2007;26:1121–1126. doi: 10.1007/s00299-006-0298-z. [DOI] [PubMed] [Google Scholar]
  30. Zhang L, French R, Langenberg W G, Mitra A. Accumulation of barley stripe mosaic is significantly reduced in transgenic plants expressing a bacterial ribonuclease. Transgenic Res. 2001;10:13–19. doi: 10.1023/A:1008931706679. [DOI] [PubMed] [Google Scholar]

Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

RESOURCES