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Isolation, selection and characterization of rhizobacteria with potential for growth promotion in Araucaria angustifolia

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Author(s):
Carlos Marcelo Ribeiro
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:
Elke Jurandy Bran Nogueira Cardoso; Fernando Dini Andreote; Lucas Carvalho Basilio de Azevedo
Advisor: Elke Jurandy Bran Nogueira Cardoso
Abstract

Araucaria angustifolia is a plant species of vital importance for the maintenance of the Mixed Ombrophylous Forest (Araucaria Forest) ecosystem in which it occurs naturally. Araucaria has a high socio-economic and environmental value, and due to its intense predatory exploitation, it is critically endangered. The destruction of this precious plant species would cause serious consequences to the ecosystem, which could involve a reduction of the diversity of many animals, plants and microorganisms, which are closely associated and reveal a great dependence on A. angustifolia. Studies involving plant growth promoting rhizobacteria (PGPR) and their characterization have never before been performed in A. angustifolia. The objective of this study was to isolate, select and characterize rhizobacteria with potential for growth promotion and biocontrol of plant pathogenic fungi in A. angustifolia. The isolates were selected by means of their potential to produce indole acetic acid (IAA), to solubilize inorganic phosphate, to produce phosphatase, and for asymbiotic N2 fixation. The latter was estimated by analyzing the accumulation of nitrogen in the culture medium and by the acetylene reduction assay (ARA). Rhizobacteria were also selected through the evaluation of indirect action mechanisms such as siderophore production and antagonism to Fusarium oxysporum, Cylindrocladium candelabrum and C. pteridis, all pathogens of trees. The most promising isolates were characterized by analyzing the fatty acid methyl ester (FAME), by BOX-PCR and partial sequencing of the 16S rRNA gene. Ninety seven rhizobacteria were isolated and subjected to the phenotypic tests. All isolates presented at least one positive feature, characterizing them as potential PGPR, except for the production of chitinases. Among these isolates, 18 produced IAA, 27 were able to solubilize inorganic phosphate, 37 were positive for siderophores and 83 were phosphatases producers. Regarding the asymbiotic N2 fixation, 20 isolates were effective in the pellicle formation in nitrogen free culture medium, however, none differed significantly from the control, when analyzed for the accumulation of N in the culture medium. Furthermore, three isolates were able to reduce acetylene to ethylene. Forty five isolates were endospore formers and antagonistic to F. oxysporum. The isolates B4, B15, B27, B35, B36, B42 and RISP2, characterized as Bacillus spp. stood out as good antagonists of the evaluated plant pathogens. We also observed the presence of strains with multiple mechanisms of action. Genotypic analysis of the most promising isolates by the BOX-PCR technique showed the formation of two major groups, expressing clearly the grouping of isolates forming endospores. Both techniques, FAME and PCR, resulted in the same grouping of the most effective isolates as belonging to Bacillaceae (9 isolates), Enterobacteriaceae (11) and Pseudomonadaceae (1). As far as we know, this is the first study to include the bacterium Ewingella americana among the PGPR. The results demonstrate a potential application of these rhizobacteria as biofertilizers and for biological control of plant pathogens in A. angustifolia. (AU)

FAPESP's process: 08/01841-0 - Diversity of fluorescent Pseudomonas spp. and Bacillus spp. in the Araucaria angustifolia rhizosphere
Grantee:Carlos Marcelo Ribeiro
Support Opportunities: Scholarships in Brazil - Master