Application of pectic zymogram in the identification and genetic variation of Fusarium species
Article Sidebar
Main Article Content
Abstract
The genus Fusarium is a species rich genus. Different media are required to study the
morphological characters and classify different species, and this is a time consuming technique.
Molecular and biochemical techniques have been used for identifying fungi over the last two
decades, including pectic zymograms which have been used to characterize different fungi. In
this study pectic zymograms were used to identify species and form species of Fusarium
isolates. Fusarium isolates (318) were obtained from different areas and hosts in Isfahan
Province, Iran. Isolates were identified to species based on morphological characters. A pectic
enzyme solution was prepared for each isolate using liquid media containing citrus pectin as the
sole carbon source. Electrophoresis was performed using acrylamide gel containing 0.2% citrus
pectin as the enzyme substrate. The gels were incubated in 0.1M malic acid before staining
overnight in 0.02% ruthenium red, to visualize enzyme electrophoretic patterns. Several
zymogram phenotypes were obtained for polygalacturonase and pectin esterase. In total, 12
zymogram patterns were determined for the 318 isolates tested. The results showed that there is
considerable intraspecific variation in Fusarium species. There were 3, 5 and 2 zymogram
electrophoretic patterns for Fusarium oxysporum, F. solani and F. culmorum respectively.
However, there were only one zymogram pattern for F. subglutinans and also one for F.
equiseti. Although the intraspecific variation based on pectic zymograms was not correlated to
the form species of Fusarium, species of Fusarium could be distinguished using this technique
as there was no common zymogram pattern among species.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Balali, G.R. (1996). Genetic variation of Rhizoctonia solani AG-3 in South Australia (Ph.D. thesis). Adelaide, South Australia.
Balali, G.R. and Kowsari. M. (2004). Pectic zymogram variation and pathogenicity of Rhizoctonia solani AG-4 to bean (Phaseoulus vulgaris) isolates in Isfahan, Iran. Mycopathologia 158: 377-384.
Balali, G.R., Kasalkheh, M. and Kowsari, M. (2000). Using pectic zymogram as a criterion to study variation and pathogenicity to field isolates of Rhizoctonia solani. 3rd International Symposium on Rhizoctonia, Taiwan. (Abstract).
Booth, C. (1975). The present status of Fusarium taxonomy. Annual Review of Phytopathology 13: 83-93.
Booth, C. (1977). Fusarium. Commonwealth Mycologia Institute. England.
Burgess, L.W., Nelson, P.E. and Summerbell, B.A. (1989b). Variability and stability of morphological characters of Fusarium oxysporum isolated from soils in Australia. Mycologia 81: 818-822.
Burgess, L.W., Nelson, P.E. and Toussoun, T.A. (1989a). Stability of morphological characters of Fusarium nygamai. Mycologia 81: 480-482.
Butler, E.E. (1980). A method for long-time culture storage of Rhizoctonia solani. Phytopathology 70: 820-821.
Cruickshank, R.H. (1983). Distinction between Sclerotinia species by their pectic zymograms. Transactions of the British Mycological Society 80: 117-119.
Cruickshank, R.H. and Pitt, J.I. (1987). Identification of species in Penicillium subgenus Penicillium by enzyme electrophoresis. Mycologia 79: 614-620.
Cruickshank, R.H. and Wade, G.C. (1980). Detection of pectic enzymes in pectin-acrylamid gels. Analytical Biochemistry 107: 177-181.
Deacon, J.W. (1997). Modern Mycology. Blackwell Science. London.
Eley, A.R. (1996). Microbial Food Poisoning. Chapman and Hall, New York.
Elias, K.S. and Schneider, R.W. (1991). Vegetative compatibility groups of Fusarium oxysporum f.sp. lycopersici. Phytopathology 81: 159-162.
Fernandez, N., Patino, B. and Vazauez, C. (1993). Pectic degrading enzymes secreted by six isolates of Fusarium oxysporum. Mycologia research 97: 461-466.
Hansen, H.N and Smith, R.E. (1932). The mechanisms of variation in imperfect fungi: Botrytis cinerea. Phytopathology 37: 369-371.
Kistler, H.C. (1997). Genetic diversity in the plant -pathogenic fungus Fusarium oxysporum. Phytopathology 87: 474-479.
Kleiner, D.E. and Stetler-Stevenson, W.G. (1994). Quantitative Zymography: Detection of picogram quantities of gelatinases. Analytical Biochemistry 218: 325-329.
Klittich, C.J., Leslie, J.F., Nelson, P.E. and Marasas, W.F.O. (1997). Fusarium thapsinum (Gibberella thapsina): A new species in section Liseola from sorghum. Mycologia 89: 643-652.
Leber, T.M. and Bakwill, F.R. (1997) Zymography: A single-step staining method for quatitation of proteolytic activity on substare gels. Analytical Biochemistry 294: 24-28.
McDonald, B.A. (1997). The population genetics of fungi: Tools and techniques. Phytopathology 87: 448-453.
Nelson, P.E., Toussoun, T.A. and Marasas, W.F.O. (1983). Fusarium species an illustrated manual for identification. The Pennsylvania State University Press, London.
Onyike, N.B.N., Nelson, P.E. and Marasas, W.F.O. (1991). Fusarium species associated with millet grain from Nigeria, Lesotho, and Zimbabwe. Mycologia 83: 708-712.
Palm, M.E., Gams, W. and Nirenberg, H.I. (1995). Plectosporium, a new genus for Fusarium tabacinum, the anamorph of Plectosphaerella cucumerina. Mycologia 87: 397-406.
Pegg, K.G., Shivas, R.G., Moore , N.Y. and Bentley, S. (1995). Characterization of a unique population of Fusarium oxysporum f.sp. cubense causing fusarium wilt in cavendish bananas of carnavon, western Australia. Australian Journal of Agricultural Research 46: 167-178.
Phelps, D.C., Nemec, S., Baker, R. and Mansell, R. (1990). Immunoassay for naphtazarin phytotoxins produced by Fusarium solani. Phytopathology 80: 298-302.
Samak, D.A. and Leong, S.A. (1989). Disease development in Cucurbita maxima (squash) infected with Fusarium solani f.sp. cucurbitae. Canadian Journal of Botany, 87: 34863489.
Sangalang, A.E., Summerbell, B.A., Burgess, L.W. and Backhouse, D. (1995). Taxonomy of Fusarium: characterization of Fusarium avenaceum subsp. aywerte and Fusarium avenaceum subsp. nurragi. Mycological Research 99: 287-290.
Shephard, G.S., Thiel, P.G., Sydenham, E.W. and Snijman, P.W. (1995). Toxicokinetics of mycotoxin fumonisim B-2 in rats. Food and Chemical Toxicology, 33: 591-595.
Snyder, W.C. and Hansen, H.N. (1941). The effect of light on taxonomic characters in Fusarium. Mycologia 33: 580-591.
Survelyer, P.S., Adams, G.C., Lezzoni, A.F. and Jones, A.L. (1995). Isozyme detection and variation in Leucostoma species from Prunus and Malus. Mycologia 87: 471-482.
Sweetingham, M.W., Cruickshank, R.H. and Wong, D.H. (1986). Pectic zymograms and taxonomy and pathogenicity of the Ceratobasidiaceae. Transactions of the British- Mycological Society 86: 305-311.
Szeczi, A. (1990a). Analysis of pectic enzyme zymograms of Fusarium species . I- Fusarium lateritium and related species. Journal of Phytopathology 128: 75-83.
Szeczi, A. (1990b). Analysis of pectic enzyme zymograms of Fusarium species. II-Comparison of polygalacturonase zymograms of Fusarium culmorum and Fusarium graminearum . Journal of Phytopathology 130: 188-196.
Yang, H.A. (1993). Genetics of strains of Rhizoctonia solani Kuhn associated with bare- patch disease of cereals (Ph D. thesis). University of Western Australia.