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Protocol establishment to control the manifestation of endophytic bacteria in the process of Eucalyptus in vitro multiplication

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Author(s):
Gabriela Ferraz Leone
Total Authors: 1
Document type: Master's Dissertation
Press: Piracicaba.
Institution: Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC)
Defense date:
Examining board members:
Marcilio de Almeida; Fernando Dini Andreote; Fernando Angelo Piotto
Advisor: Marcilio de Almeida
Abstract

The in vitro bacterial presence in plant cultivation, usually is related to contamination, resulting in the elimination of the microplants with high commercial value. However, this presence can be characterized as \"manifestations\" arising from possible physical or nutritional stresses, from microbial endophytic that actively colonize plant tissues, and do not necessarily affect plant development. Therefore, the purpose of this study was to control the bacterial manifestation, preserving in vitro developing of Eucalyptus cloeziana microstumps, by adding to the culture medium, the antibiotics gentamicin, ciprofloxacin, rifampicin, timentin (alone or combined) and biocide + chloromethyl isothiazolinone methyl-isothiazolinone. The monitoring work was conducted through weekly evaluations, to verify the possible influence of antimicrobial agents in the plant morphophysiology and the efficient control of bacterial manifestations. Therefore, were evaluated parameters related to the increase in dry weight, number of buds, content of nutrient accumulation, histological and histochemical the microstumps. DGGE technique was used to check possible fluctuations of endophytic bacterial communities in microstumps. All analyzes were performed at 15 and 30 days after the addition of antimicrobial agents in the culture medium, except for the nutritional assessment that was made only at the end of the experiment (30 days). The results allow to infer that over the 30 days of culture, the antibiotic gentamicin, resulted in the microstumps development decrease and was not effective in controlling bacterial manifestations. The antibiotics rifampicin and ciprofloxacin although were not effective in controlling the bacterial manifestation, affected the development of microstumps. It was concluded that the biocide chloromethyl-methyl-Isothiazolinone + Isothiazolinone effectively controlled the bacterial manifestations without modifying the morphophysiology of the microstumps, well as the timentin, which besides controlling the manifestations, provided greater vigor of microstumps due to better assimilation of nutrients. (AU)

FAPESP's process: 11/15861-6 - Establishment of a protocol to control expression of endophytic bacteria in the process of in vitro multiplication of Eucalyptus
Grantee:Gabriela Ferraz Leone
Support Opportunities: Scholarships in Brazil - Master