Phytopathogenic fungi during the penetration and colonization process are capable of secreting several enzymes, which enable infection of the host live tissue, acting on the degradation of wax, cuticle and cell walls. The ability of a pathogenic agent to produce enzymes or not can determine the severity degree of a disease. In this study, 33 isolates of Colletotrichum gloeosporioides related to anthracnose and blister spot on coffee trees were evaluated for their ability to produce hydrolytic enzymes (amylase, lipase, protease, laccase, pectinase and cellulase) and specific cell wall degrading enzymes “CWDEs” (polygalacturonase, polymethylgalacturonase and pectin-lyase), as well as their relationship with the pathogenicity/aggressiveness of isolates. For all isolates of C. gloeosporioides, extracellular enzymes could be detected, except cellulases. Isolates I-9 and I-24 produced the highest levels of extracellular enzymes, as well as CWDEs. They also had the highest disease intensity indexes, suggesting a relationship between enzymes and aggressiveness of the isolates.
Blister spot (Colletotrichum gloeosporioides) is now widespread in most coffee producing states of Brazil, becoming a limiting factor for production. The lack of data relating to the reproduction of typical symptoms (light green, oily patches) leaves a gap within the pathosystem, forcing the search for new methodologies for monitoring the disease. Monitoring of genetically modified organisms has proven to be an effective tool in understanding the host x pathogen interactions. Thus, the present study was carried out to evaluate the effectiveness of two systems of genetic transformation in obtaining mutants using the gfp reporter gene.Using the two transformation systems (PEG and electroporation) revealed the efficiency of both, confirmed by fluorescence microscopy and resistance to the antibiotic hygromycin-B, when incorporated into the culture medium. The fungus maintained its cultural and morphological characteristics when compared to wild strains. When inoculated on coffee seedlings, it was found that the pathogenicity of the processed isolates had not changed.
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