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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.1986.tde-20240301-150636
Document
Author
Full name
Amalia Del Rosaria Baraibar Lucas
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1986
Supervisor
Title in Portuguese
Caracterização e seleção de Rhizobium phaseoli para a produção de inoculantes comerciais
Keywords in Portuguese
BACTÉRIAS FIXADORAS DE NITROGÊNIO
INOCULANTES
PRODUÇÃO
RIZÓBIO
SELEÇÃO
Abstract in Portuguese
Foram conduzidos estudos de antagonismo entre Streptomyces isolados de turfas e estirpes de R. phaseoli a partir dos quais obtiveram-se variantes das estirpes CO5, SEMIA 487 e CIAT 255 que cresciam nos halos de inibição formados. Com estas variantes, as culturas matrizes e outras estirpes de R. phaseoli, foi proposta uma sequência de testes que inclue: resistência a antibióticos e fungicidas, produção de bacteriocinas, tolerância à acidez, eficiência simbiótica, competição nodular e sobrevivência na turfa. Os isolados C05 I, C05 II e C05 III, SEMIA 487-2 e a estirpe 1899 apresentaram altos níveis de resistência à estreptomicina, cloranfenicol, eritromicina e tetraciclina assim como benomyl, thiram e captan e cresceram em melo ácido (pH = 4,3). Nenhuma das estirpes testadas apresentou atividade bacteriocinogênica. As estirpes C05 e SEMIA 487 foram ineficientes; o isolado C05 II demonstrou superioridade nos parâmetros simbióticos estudados apresentando a maior eficiência nodular, foi altamente competitivo para a formação de nódulos na presença de estirpes de Rhizobium nativas e frente a C05 matriz, e igualmente competitivo em relação à estirpe 1899. As estirpes matrizes C05 e SEMIA 487 nodularam tardiamente com nódulos pequenos e escassos, 1899 e SEMIA 487-2 foram de nodulação precoce e ciclo curto (até floração) enquanto que, C05 II destacou-se pelo ciclo de nodulação mais prolongado. Discute-se a validez da persistência nodular como critério de seleção de estirpes para Phaseolus vulgaris. A estirpe CO5 não sobreviveu satisfatoriamente nas turfas não irradiadas, sendo superada pelos isolados C05 I, C05 II e C05 III e pela estirpe 1899. A irradiação da turfa com radiações gama (2,5 Mrad) controlou o crescimento dos contaminantes remanescentes e permitiu boa sobrevivência do R. phaseoli durante os 60 dias de armazenamento dos inoculantes a 28°C. Recomenda-se o isolado C05 II para a produção de inoculantes para feijoeiro. Para certos casos, a estirpe 1899 poderá ser incluída junto à C05 II em inoculantes polivalentes porque a primeira apresenta um ciclo de nodulação precoce. Discute-se a possibilidade de irradiação da turfa com doses acima de 2,5 Mrad e uso de estirpes com resistência a antibióticos e fungicidas como alternativa para a produção de inoculantes de maior qualidade no Brasil.
Title in English
Characterization and selection of Rhizobium phaseoli for commercial inoculant production
Abstract in English
Antagonism lnteraction studies between Streptomyces from peat and Rhizobium phaseoli strains were conducted in agar medium. Colonies from strains C05, SEMIA 487 and CIAT 255, growing in the zone of inhibition formed in presence of actinomycetes (Streptomyces) isolates, were picked up and cultured. A sequence of tests was proposed with those isolates with the aim to select better strains. Intrinsic antibiotic and fungicide resistance, bacteriocin production, acid tolerance, effectiveness, competitiveness for nodule formation and survival ln the peat were considered as the main propertles for the screening. lsolates COS I, C05 II, C05 III, SEMIA 487-2 and the strain 1899 presented high resistance level to streptomycin, chloranphenicol, eritromycin and tetracycl in as well as to benomy, thiram or captan, with normal growth in acid media (pH = 4.3). Mother strains C05 and SEMIA 487 were inefficient and isolate COS II was superior for all the studied parameters, with the highest nodular efficiency. It was highly competitive for nodule formation in presence of the native strains from soil or the mother strain CO5 and was a competitive as strain 1899. The pattern of nodulation (nitrogenase activity and nodule size) was different among strains. Strains CO5 and SEMIA 487 formed their nodules mostly in a later period and they were very small and sparse. Strain 1899 and the isolate SEMIA 487-2 nodulated earlier, with a short period of activity (until flowering); isolates C05 II presented a longer period of nodule activity. The importance of the nodule persistance as an strain selection parameter for R. phaseoli is discussed. The mother culture CO5 showed a low survival ability in the unsterilized carrier. lrradiation of the peat with gamma rays at rates of 2.5 Mrad controlled the growth of the remaining contaminants and allowed a good survival of R. phaseoli during the period of sixty days of storage at 28°C. Due to those intrinsic characteristics, isolate COS II is recommended for the inoculaht production for Phaseolus vulgaris. Mixtures in equal parts of 1899 and CO5 II are recommended for some specific ecological situations. The use of irradiated peat associated to the use of antibiotic and fungicide resistant strains is discussed as an alternative to improve the quality of Brazilian inoculants.
 
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