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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