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| Author(s): |
Maria Carolina Quecine
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Piracicaba. |
| Institution: | Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC) |
| Defense date: | 2010-03-10 |
| Examining board members: |
Aline Aparecida Pizzirani kleiner;
João Lucio de Azevedo;
Carlos Alberto Labate;
Paulo Teixeira Lacava;
Gabriel Padilla Maldonado
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| Advisor: | Aline Aparecida Pizzirani kleiner |
| Abstract | |
Sugarcane is one of the most important Brazilian crops. Its importance is increasing due to the gradual substitution of fossil fuel for renewable and cleaner energy sources, such as ethanol Consequently, the biotechnological aspects that mediate sugarcane-bacterium interactions must be evaluated; studies from tropical countries are incipiente Bacterium-plant interactions can often improve the growth and fitness of associated plants. One of these beneficial bacteria, Pantoea agglomerans, has been isolated from many vegetal species. The Pseudomonas genus also has a record of biotechnological exploration due to its wide range of antibiotic production. These compounds can be used on pest and plant-pathogen control. For these reasons, P. agglomerans, strain 33.1, previously isolated from Eucalyptus grandis and a known eucalyptus growth-promoter, was evaluated about regarding the biotechnological aspects of sugarcane-bacterial association. 33.1 strain was able to promote sugarcane growth and also to induce the production of resistance proteins for inoculated plants. The plant-hormone and phosphatase production by 33.1 were associated with sugarcane growth-promotion bacterial mechanisms. An integrative plasmid, pNKGFP, was constructed and used to transform 33.1. The marked strain was monitored during sugarcane association. The sugarcane cross-colonization by 33.1:pNKGFP was confirmed by fluorescence microscopy, re-isolation and quantitative PCR (qPCR), suggesting that 33.1 should be used in association with other crops. 33.1 was genetically modified to produce the Cry protein. The partial biocontrol of Diatraea saccharalis, the sugarcane borer, by the endophytic strain expressing heterologously the cry1a7c gene, was confirmed in bioassays using artificial diet and sugarcane stalks. Several antibiotics and siderophores produced by P. fluorescens Pf-5 were tested against Fusarium spp.; including isolates of F. verticillioides responsible for the sugarcane Pokkah boeng disease. By deriving site-directed mutants of Pf-5, with single and multiple mutations, in the bio-synthetic gene clusters of the antifungal compounds, it was proved that 2,4-diacetyl-phoroglucinol and rhizoxin are the major compounds responsible for the Fusarium spp. suppression. The fusaric acid effects on Pf-5 antibiotic production was proved by HPLC and RT-qPCR. (AU) | |