Genetic transformation with the gfp gene of Colletotrichum gloeosporioides isolates from coffee with blister spot

Cecilia Armesto, Fernanda Gonçalves Martins Maia, Mário Sobral de Abreu, Antonia dos Reis Figueira, Bruno Marques da Silva, Fernando Pereira Monteiro
2012 Brazilian Journal of Microbiology  
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
more » ... 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. Transformation of Phomopsis viticola with the green fluorescent protein. Mycologia, 101 (6), 853-858. 2. Armesto, C.; Martins-Maia, F.G.; Abreu, M.S.; Reis, A.F.; Alencar, N.E. (2011). Produção e regeneração de protoplastos de Colletotrichum gloeosporioides em diferentes condições de cultivo. Biosci. J., 27 (4), 597-602. 3. Bigs, J.; Geider, K. (1999). Moviment of Erwinia amylovora in host plants and bacterial sorbitol and sucrose metabolism assayed with the green fluorescent protein. New aspects of resistance research on cultivaded plants: bacterial diseases. Springer Verlag, Berlin. 4. Bitancourt, A.A. (1958). As manchas da folha do cafeeiro. Biológico., 24 (17), 191-201. 5. Chalfie, M.; Tu, Y;, Euskirchen, G.; Ward, W.W.; Prasher, D.C. (1994). Green fluorescent protein as a marker for gene expression. Science. 263 (11), 802-805. 6. Chen, N.; Hsiang, T.; Goodwin, P. H. (2003). Use of green fluorescent protein to quantify the growth of Colletotrichum during infection of tobacco.
doi:10.1590/s1517-83822012000300050 fatcat:ooe3cvmkjbci7aurfzhokhnaye