In vitro characterization andin vivo detection ofRigidoporus lignosus,the causal agent of white root disease inHevea brasiliensis,by ELISA techniques

The aim of these studies is to develop a method for early detection ofRigidopoms lignosus (Basidiomycete, Polyporaceae), the causative agent of white root disease of rubber tree. Two polyclonal sera were produced against soluble mycelial proteins of twoR. lignosus isolates, one from Africa (FCI2), the other from Asia (FID2). The specificity of the antisera was tested using isoelectric focusing (IEF)/Western-blot and DAS-ELISA. The two sera recognized all 20R. lignosus isolates from various geographical origins. The banding patterns obtained by Western-blot enabled a distinction to be made between isolates from Africa and those from Asia. In DAS-ELISA and Western-blot analyses, strong cross reactions were observed withR. ulmarius. Only slight reactions were observed in Western-blot analysis toR. lineatus andP. noxius, both causative agents of root rot inHevea. These cross reactions were not observed under our DAS-ELISA analysis conditions. Finally, no cross reactions were obtained with 9 otherPolyporaceae orHevea root pathogen species. The sensitivity threshold of the DAS-ELISA method was 5 ng ml^–1 ofR. lignosus protein. An initial approach to using the DAS-ELISA test for the detection ofR. lignosus in infected plants was carried out on artificially inoculated root samples. The DAS-ELISA protocol enabled detection ofR. lignosus in the root systems of diseased plants. Moreover, no cross reaction was observed with healthy plant extracts.