Polyclonal antibodies anti-PfP IgG and anti-PfM IgG and monoclonal antibody MAb29 were prepared to detect a quarantine pathogen of strawberry, Phytophthora fragariae. Laboratory rabbits and mice were immunized using purified and unpurified protein extracts from the mycelial mass of the pathogen. All prepared antibodies were genus-specific, therefore Phytophthora cactorum was also detected. Except for Pythium ultimum, the antibodies did not cross-react with other pathogenic fungi, such as Botrytis cinerea, Colletotrichum acutatum, Fusarium sp., Verticillium albo-atrum. PTA-ELISA was used to test the antibodies. P. fragariae was detected in artificially infected strawberries (cultivars Elsanta, Kama and Vanda) by means of PTA-ELISA, immunoprinting and dot blot. Detection of the pathogen was optimal in undamaged roots or roots with necrotic tips only. At a later stage of infection, when whole roots were necrotic, the crown was more suitable for successful detection. To detect P. fragariae at the early stages of infection it is recommended to use at least two of the three mentioned immunotechniques.
Four polyclonal and two monoclonal antibodies were prepared and tested to detect a quarantine pathogen of strawberry – Colletotrichum acutatum. Only one of them, polyclonal IgG K91, was sensitive enough to recognize the pathogen. This antibody was genus-specific and did not cross-react with several other fungal pathogens of strawberry (Phytophthora fragariae, P. cactorum, Verticillium albo-atrum, Botrytis cinerea, Pythium ultimum). Four techniques, PTA-ELISA, dot blot, immunoprint and immunofluorescent microscopy were used to test the specifity and sensitivity of antibodies. After artificial infection of strawberry (cvs Elsanta, Vanda, and Kama), Colletotrichum acutatum was detected by PTA-ELISA, dot blot and immunoprint in roots, crowns, petioles and fruits in the latent stage of the disease. For reliable detection in the latent stage it is recommended to use at least two of the mentioned techniques.
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