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1. Paula Garcia Meirelles. Fusarium verticillioides : caracterização molecular e detecção em milho através de ensaio imunoenzimático.

Degree: 2005, Universidade Estadual de Londrina

Fusarium verticillioides is a primary corn pathogen and the main fumonisin producer. This fungus can cause significant economical losses for the farmers, grain processors, animal producers and risk for human and animal health. The aims of this study were to develop an enzyme-linked immunosorbent assay (ELISA) to detect F. verticillioides in corn samples and evaluate the correlation between this methodology and traditional methods of fungal and fumonisin detection, as well as analyse polymorphism in FUM6 gene, involved in fumonisins biosynthesis. The polymorphisms observed were then correlated to fumonisin B1 (FB1) production. Exoantigens from 8 F. verticillioides strains were obtained for the development of an indirect competitive ELISA. The exoantigens from 3 strains (97K, 113F and 162A) were selected for polyclonal antibody production, due to the highest protein concentration and high number of visible protein bands by denaturing polyacrilamide gel electrophoresis (SDS-PAGE). Antiserum against F. verticillioides 97K exoantigens, which showed the highest titre in indirect ELISA (1:12.800), was chosen for further analyses. Forty freshly harvested corn samples were analysed for F. verticillioides exoantigens, as well as for total mould count, ergosterol and fumonisin levels in order to evaluate the relationship between these parameters. In addition, F. verticillioides was grown in brain heart infusion (BHI) broth, in order to evaluate the correlation between biomass and exoantigens. The electrophoretic profile of the exoantigens produced for ELISA development showed protein bands with molecular mass ranging from 17 to 170 kDa. There was no significant correlation between exoantigens and total mould count, Fusarium sp. count and fumonisina levels. Otherwise, the correlation coefficient between exoantigens and ergosterol content was 0.52 and between biomass and F. verticillioides exoantigens in BHI broth was 0.84. Three strains of F. verticillioides showed different electrophoretic profile from the others, demonstrating that there is polymorphism for fum6 locus. Strain 97L showed polymorphism at the region corresponding to P450 conserved domain while strains 97L and 119Fa showed polymorphism at FAD_binding 1 domain. The polymorphism at P450 domain is not apparently associated with high or low levels of FB1 synthesis, as it produced intermediate levels of FB1 (1050 µg/g), however the strains which showed polymorphism at FAD_binding 1 domain presented high levels of FB1 (2643 and 2814 µg/g respectively, for strains 97L and 119Fa). The results suggest that the ELISA with exoantigens has potential for Fusarium sp. detection in corn samples and that FUM6 gene can be a molecular marker for high fumonisin producer strains of F. verticillioides.

Fusarium verticillioides é um patógeno primário de milho e principal produtor de fumonisinas. Este fungo pode causar perdas econômicas significativas para produtores e processadores de grãos, criadores de animais, além de representar sérios riscos à saúde humana e animal. Este…

Advisors/Committee Members: Elisabete Yurie Sataque Ono ., Elisa Yoko Hirooka, Maria Helena P. Fungaro.

Subjects/Keywords: Milho; Fusario; Fusarium verticillioides; Micotoxinas; Fumonisinas; Corn; Mocotoxins; Fusarium

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APA (6th Edition):

Meirelles, P. G. (2005). Fusarium verticillioides : caracterização molecular e detecção em milho através de ensaio imunoenzimático. (Thesis). Universidade Estadual de Londrina. Retrieved from http://www.bibliotecadigital.uel.br/document/?code=vtls000111970

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Meirelles, Paula Garcia. “Fusarium verticillioides : caracterização molecular e detecção em milho através de ensaio imunoenzimático.” 2005. Thesis, Universidade Estadual de Londrina. Accessed August 22, 2019. http://www.bibliotecadigital.uel.br/document/?code=vtls000111970.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Meirelles, Paula Garcia. “Fusarium verticillioides : caracterização molecular e detecção em milho através de ensaio imunoenzimático.” 2005. Web. 22 Aug 2019.

Vancouver:

Meirelles PG. Fusarium verticillioides : caracterização molecular e detecção em milho através de ensaio imunoenzimático. [Internet] [Thesis]. Universidade Estadual de Londrina; 2005. [cited 2019 Aug 22]. Available from: http://www.bibliotecadigital.uel.br/document/?code=vtls000111970.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Meirelles PG. Fusarium verticillioides : caracterização molecular e detecção em milho através de ensaio imunoenzimático. [Thesis]. Universidade Estadual de Londrina; 2005. Available from: http://www.bibliotecadigital.uel.br/document/?code=vtls000111970

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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