Serine Proteinases Secreted by Two Isolates of the Fungus Alternaria solani


Fungus, trypsin-like, subtilisin-like, proteinase.

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Tatiana A. Valueva, Natalia N. Kudryavtseva, Ekaterina L. Gvozdeva, Alexis V. Sof’in, Natalia Yu. Il’ina, Marina A. Pobedinskaya, & Sergei N. Elansky. (2013). Serine Proteinases Secreted by Two Isolates of the Fungus Alternaria solani. Journal of Basic & Applied Sciences, 9, 105–115.


It is well-known Alternaria solani Sorauer is the causative agent of alternariosis. In this paper, serine proteinases secretion by two genetically related isolates of the fungus, collected from potato and tomato plants grown in central Russia have been studied. The data clarify functions of these enzymes in the process of pathogenesis in which they can play a pivotal role. Also, the data should allow classifying Alternaria’s strains more precisely. It was found that the two isolates produced trypsin-like and subtilisin-like proteinases during growth both in synthetic culture medium and in medium containing heat-stable vegetable proteins. There were significant differences in the influence of the environment on the serine proteinase secretion by the potato and tomato isolates of A. solani. The proportion of such serine proteinases as trypsin-like and subtilisin-like enzymes depends on the composition of the growth medium, especially on the available organic nitrogen form, as well as features both of the pathogenic fungus and of the host plant. So, the tomato isolate demonstrated weak growth and low level or absence of serine proteinase excretion on cultivation with the medium containing proteins extracted from potato tubers and pea seeds. The potato isolate secreted many more serine proteinases, among which the trypsin-like enzymes dominated. Our data suggest that the tomato isolate, when grown on medium with proteins extracted from potato tubers, lost pathogenicity and became to behave as a saprophyte, while the potato isolate retained its pathogenic properties on growth on any tested medium.


Rotem J. The Genus Alternaria: Biology, Epidemiology and Pathogenicity. St Paul Minnesota USA: The American Phytopathological Society 1994.

Simmons EG. Alternaria an identification manual. Utrecht the Netherlands: CBS Fungal Biodiversity Center 2007.

Pscheidt JW, Stevenson WR. Early blight of potato and tomato: A Literature Review. Madison WI USA: College of Agriculture & Life Sciences 1986.

Orina AC, Gannibal PhB, Levitin MM. Specific diversity, biological characters and geography of Alternaria fungi associated with Solanaceous plants. Mycol Phytopath (Russia) 2010; 44(1): 150-9.

Lourenço V, Moya A, Gonzalez-Candelas F, Carbone I, Maffia LA, Mizubuti ESG. Molecular diversity and evolutionary processes of Alternaria solani in Brazil inferred using genealogical and coalescent approaches. Phytopathology 2009; 99 (7): 765-74.

Tunlid A, Talbot NJ. Genomics of parasitic and symbiotic fungi. Curr Opin Microbiol 2000; 5: 513-9.

Soanes DM, Alam I, Cornell M, et al. Comparative genome analysis of filamentous fungi reveals gene family expansions associated with fungal pathogenesis. PLoS ONE 2008; 3: e2300. pone 0002300

Movahedi S, Heale JB. Purification and characterization of an aspartic proteinase secreted by Botrytis cinerea Pers. ex Fries in cuticle and infected carrots. Physiol Mol Plant Path 1990; 36: 303-24.

Ball AM, Ashby AM, Daniels MJ, Ingram DS, Johnstone K. Evidence for the requirement of extracellular protease in the pathogenic interaction of Pyrenopeziza brassicae with oilseed rape. Physiol Mol Plant Path 1991; 38 (1): 147-61.

Paris R, Lamattina L. Phytophthora infestans secretes extracellular proteases with necrosis inducing activity on potato. Eur J Plant Path 1999; 105: 753-60.

Sabotic J, Trcek T, Popovic T, Brzin J. Basidiomycetes harbour a hidden treasure of proteolytic diversity. J Biotech 2007; 128(2): 297-307.

Valueva TA, Mosolov VV. Role of inhibitors of proteolytic enzymes in plant defense against phytopathogenic microorganisms. Biochemistry (Moscow) 2004; 69(8): 1305-9.

Pavlukova EB, Belozersky MA, Dunaevsky YE. Extracellular proteolytic enzymes of filamentous fungi. Biochem (Moscow) 1998; 63(9): 899-28.

Monod M, Capoccia S, Léchenne B, Zaugg C, Holdom M, Jousson O. Secreted proteases from pathogenic fungi. Int J Med Microbiol 2002; 292(2): 405-19.

Page MJ, Di Cera E. Serine peptidases: classification, structure and function. Cell Mol Life Sci 2008; 65: 1220-36.

Hu G, St Leger RJ. A phylogenomic approach to reconstructing the diversification of serine proteases in fungi. J Evol Biol 2004; 17: 1204-14.

Dubovenko AG, Dunaevsky YE, Belozersky MA, Oppert B, Lord JC, Elpidina EN. Trypsin-like proteins of the fungi as possible markers of pathogenicity. Fungal Biol 2010: 114 (1): 151-9.

Elansky S, Pobedinskaya M, Kokaeva L, Statsyuk N, Alexandrova A. Molecular identification of the species composition of Russian isolates of pathogens, causing early blight of potato and tomato. Proceedings of the 13th EuroBlight workshop. PPO-Special Report 2012; 15: 151-6.

Lisinska G, Leszczynski W. Potato Science and Technology. London, New York: Elsevier Applied Science Publishers Ltd 1989; p. 391.

Igbasan FA, Guenter W, Slominski BA. Field peas: Chemical composition and energy and amino acid availabilities for poultry. Can J Anim Sci 1997; 77(2): 293-300.

Sharma KD, Karki S, Thakur NS, Attri S. Chemical composition, functional properties and processing of carrot – a review. J Food Sci Technol 2012; 49(1): 22-32.

Charney J, Toarelli RM. A colorimetric method for the determination of the proteolytic activity of duodenal juice. J Biol Chem 1947; 171: 501-5.

Erlanger DF, Kokowsky N, Cohen W. The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys 1961; 95(2): 271-8.

Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem 1980; 102(2): 196-202.

Dunaevsky YE, Belyakova GA, Pavlukova EB, Belozersky MA. Influence of cultivation conditions on synthesis and secretion of proteases by the fungi Alternaria alternata and Fusarium oxysporum. Microbiol (Moscow) 1995; 64(2): 327-30.

Cohen B. Regulation of intracellular and extracellular neutral and alkaline proteases in Aspergillus nidulans. J Gen Microbiol 1973; 79: 311-20.

Pateman J.A, Kinghorn JR, Dunn E, Forbes E. Ammonium regulation in Aspergillus nidulans. J Bacteriol 1973; 114: 943-50.

Kalisz H, Pohlig G, Holzer H. Inhibition of protein phosphorylation by chloroquine. Arch Microbiol 1987; 147(2): 235-9.

Dunaevsky YE, Gruban TN, Belyakova GA, Belozersky MA. Extracellular proteinases of filamentous fungi as potential markers of phytopathogenesis. Microbiol (Moscow) 2006; 75(3): 649-52.

Valueva TA, Kudryavtseva NN, Sof’in A, Revina TA, Gvozdeva EL, Ievleva EV. Environmental conditions as a determinant of the exoproteinase composition of three phytopathogenic microorganisms. J Pathogens 2011.

Katz ME, Flynn PK, van Kuyk PA, Cheetham BF. Mutations affecting extracellular protease production in the filamentous fungus Aspergillus nidulans. Mol Gen Gen 1996; 250: 715-24.

St Leger RJ, Nelson JO, Screen SE. The entomopathogenic fungus Metarhizium anisopliae alters ambient pH, allowing extracellular protease production and activity. Microbiol 1999; 145: 2691-9.

Yike I, Rand T, Dearborn DG. The role of fungal proteinases in pathophysiology of Stachybotrys chartarum. Mycopathologia 2007; 164: 171-81.

Revina TA, Speranskaya AS, Kladnitskaya GV, Shevelev AB, Valueva TA. Subtilisin protein inhibitor from potato tubers. Biochemistry (Moscow) 2004; 69(10): 1092-8.

Revina TA, Kladnitskaya GV, Gerasimova NG, Gvozdeva EL, Valueva TA. Protein trypsin inhibitor from potato tubers. Biochemistry (Moscow) 2010; 75(1): 36-40.

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Copyright (c) 2013 Tatiana A. Valueva, Natalia N. Kudryavtseva, Ekaterina L. Gvozdeva, Alexis V. Sof’in, Natalia Yu. Il’ina, Marina A. Pobedinskaya , Sergei N. Elansky