Abstract
Polyclonal antiserum was raised against the mycelial extract ofPestalotiopsis theœ and immunoglobulin fractions were purified by ammonium sulfate fractionation and chromatography on DEAE-Sephadex. In enzyme-linked immunosorbent assay, antiserum dilution up to 1∶16000 detected homologous antigen at a 5 mg/L concentration, and at 1∶125 antiserum dilution fungal antigens could be detected at a concentration as low as 25 μg/L. In fifteen varieties of tea tested, originating from Darjeeling, UPASI and Tocklai breeding stations, absorbance values of infected leaf extracts were significantly higher than those of healthy extracts at a concentration of 40 mg/L in indirect ELISA. ELISA-positive material was detected in tea leaves as early as 12 h after inoculation withP. theœ. At antiserum dilutions up to 1∶125, the pathogen could be detected in inoculated leaf extracts up to antigen concentration of 2 mg/L. The antiserum reacted with two other isolates ofP. theœ tested but not with the antigens from mycelial extracts ofGlomerella cingulata andCorticium invisum or with extracts of tea leaves inoculated with these pathogens. The results demonstrate that ELISA can be used for early detection ofP. theœ in leaf tissues even at a very low level of infection.
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References
Alba A.P.C., Devay J.E.: Detection of cross-reactive antigens betweenPhytophthora infestans (Mont.) de Bary andSolanum species by indirect enzyme linked immunosorbent assay.Phytopath. Z. 112, 97–104 (1985).
Barbora B.C.: Latest concept on management of young tea, pp. 55–62 inField Management of Tea (S.K. Sarkar H.P. Bezbaruah, Eds). Tocklai Experimental Station, Jorhat 1988.
Barker I., Pitt D.: Detection of the leaf curl pathogens of anemones in corms by enzyme linked immunosorbent assay (ELISA).Plant Pathol. 37, 417–422 (1988).
Beckman K.B., Harrison J.G., Ingram D.S.: Optimization of a polyclonal enzyme linked immunosorbent assay (ELISA) of fungal biomass for use in studies of plant defence responses.Physiol. Mol. Plant Pathol. 44, 19–32 (1994).
Chakraborty B.N., Purkayastha R.P.: Serological relationship betweenMacrophomina phaseolina and soybean cultivarsPhysiol. Plant Pathol. 23, 197–205 (1983).
Chakraborty B.N., Saha A.: Detection and cellular location of cross reactive antigens shared byCamellia sinensis andBipolaris carbonum.Physiol. Mol. Plant Pathol. 44, 403–416 (1994a).
Chakraborty B.N., Saha A.: Accumulation of antifungal compounds in tea leaf tissue infected withBipolaris carbonum.Folia Microbiol. 39, 409–414 (1994b).
Chakraborty B.N., Basu P., Das R., Saha A., Chakraborty U.: Detection of cross reactive antigens betweenPestalotiopsis theœ and tea leaves and their cellular location.Ann. Appl. Biol. 127, 11–21 (1995).
Clausen J.: Immunochemical techniques for the identification and estimation of macromolecules, Vol. 1, Part 3 of the seriesLaboratory Techniques in Biochemistry and Molecular Biology (R.H. Burdon, P.H. Van Knippenberg, Eds). Elsevier, Oxford 1988.
Dickens J.S.W., Cook R.T.A.:Glomerella cingulata onCamellia.Plant Pathol. 38, 75–85 (1989).
Elnashaar H.M., Moore L.W., George R.A.: The use of ELISA for early quantification ofGœumannomyces graminis f.sp.tritici associated with winter wheat roots.Phytopathology 76, 1319–1322 (1986).
Harrison J.G., Barker R., Lowe R., Rees E.A.: Estimation of amounts ofPhytophthora infestans mycelium in leaf tissue by enzyme linked immunosorbent assay.Plant Pathol. 39, 274–277 (1990).
Holtz B.A., Karu A.E., Weinhold A.R.: Enzyme linked immunosorbent assay for detection ofThielaviopsis basicola.Phytopathology 84, 977–983 (1994).
Johnson M.C., Pirone T.P., Siegel M.R., Varney D.R.: Detection ofEpichlœ typhina in tall fescue by means of enzyme linked immunosorbent assay.Phytopathology 72, 647–650 (1982).
Kitagawa T., Sakamoto Y., Furumi K., Ogura H.: Novel enzyme immunoassays for specific detection ofFusarium oxysporum f.sp.cucumerinum and for general detection of variousFusarium species.Phytopathology 79, 162–165 (1989).
Linfield C.A.: A rapid serological test for detectingFusarium oxysporum f.sp.narcissi inNarcissus.Ann. Appl. Biol. 123, 685–693 (1993).
Lyons N.F., White J.G.: Detection ofPythium violœ andPythium sulcatum in carrots with cavity spots using competition ELISA.Ann. Appl. Biol. 120, 235–244 (1992).
Mohan S.B.: Evaluation of antisera raised againstPhytophthora fragariœ for detecting the red core disease of strawberries by enzyme linked immunosorbent assay (ELISA).Plant Pathol. 37, 206–216 (1988).
Mohan S.B.: Cross reactivity of antiserum raised againstPhytophthora species and its evaluation as a genus detecting antiserum.Plant Pathol. 38, 352–363 (1989).
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Chakraborty, U., Basu, P., Das, R. et al. Evaluation of antiserum raised againstPestalotiopsis theœ for the detection of grey blight of tea by ELISA. Folia Microbiol 41, 413–418 (1996). https://doi.org/10.1007/BF02815691
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DOI: https://doi.org/10.1007/BF02815691