Characterization of Rhizobia Isolated from Different Divergence Groups of Tropical Leguminosae by Comparative Polyacrylamide Gel Electrophoresis of their Total Proteins

Moreira, F. M. S.; Gillis, Monique; Pot, Bruno; Kersters, Karel; Franco, A. A.

doi:10.1016/s0723-2020(11)80258-4

Cited by 131 publications

(125 citation statements)

References 36 publications

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“…The only strain isolated from a species of the subfamily Caesalpinioideae, SEMIA 6099, was also quite similar in all ribosomal regions to SEMIA 6093, isolated from Aeschynomene americana, in the subfamily Papilionoideae. Thus, our data confirm previous results obtained in the analysis of protein profiles of 171 strains belonging to 14 legume species (Moreira et al, 1993), since no evolutionary correlation was observed between bradyrhizobial clusters and host plants.In conclusion, three ribosomal regions were analysed from a collection of strains classified as Bradyrhizobium based on morphological and physiological properties in vitro (FEPAGRO, 1999). All strains were characterized by high efficiency of N 2 fixation and had been isolated from a wide range of host tropical legumes.…”

supporting

confidence: 92%

“…van Rossum et al, 1995;Moreira, 1991Moreira, , 2000Moreira et al, 1993;Boddey & Hungria, 1997; Doignon-Bourcier et al, 1999;Chen et al, 2002). However, the diversity observed among bradyrhizobial strains is usually not reflected in 16S rRNA gene sequence analysis; therefore, few Bradyrhizobium species have been described so far.…”

Section: Discussionmentioning

confidence: 99%

“…However, the diversity observed among bradyrhizobial strains is usually not reflected in 16S rRNA gene sequence analysis; therefore, few Bradyrhizobium species have been described so far. As bradyrhizobia seem to constitute a major part of the rhizobial isolates in the tropics (Norris, 1965;Moreira, 2000) and as rhizobia might be more diverse in the tropics (Oyaizu et al, 1992;Moreira et al, 1993;Urtz & Elkan, 1996;Vinuesa et al, 1998Vinuesa et al, , 2005a, a large collection of Bradyrhizobium strains would add valuable information about the diversity of ribosomal genes within the genus.The genetic characterization in this study was performed with 119 strains from the Brazilian official culture collection SEMIA of rhizobial strains. The strains had been isolated from 33 legume species (Table 1), most from the tropics, representing nine tribes and including all three subfamilies (Supplementary Table S2).…”

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RFLP analysis of the rRNA operon of a Brazilian collection of bradyrhizobial strains from 33 legume species

Germano

Menna

Mostasso

et al. 2006

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLO

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Genetic diversity in tropical rhizobial species is still poorly known. With the aim of increasing this knowledge, three ribosomal regions of 119 strains belonging to the official Brazilian culture collection of rhizobia and classified as Bradyrhizobium based on morphological and physiological characteristics in vitro were analysed by RFLP-PCR. The strains were isolated from 33 legume species, representing nine tribes and all three subfamilies; they all form very effective N 2 -fixing nodules and 43 of them are recommended for use in Brazilian commercial inoculants as the most effective for their hosts. For the 16S rRNA gene, type and reference strains of Bradyrhizobium japonicum fell into two major clusters, joined at a level of similarity of 50 %, which included 52 strains, 90 % of which were isolated from soybean. Two other clusters, joined at a similarity of 53 %, included reference strains of Bradyrhizobium elkanii, but not USDA 76 T ; furthermore, two other major clusters were identified and all strains were clustered at a final level of similarity of only 28 %. For the intergenic spacer (IGS) between genes coding for the 16S and 23S rRNA, strains were clustered at a final level of similarity of 27 %. Reference strains of B. japonicum fell into a major group with 51 strains, 84 % isolated from soybean, with a similarity of 59 %, while strains of B. elkanii fell into another major group, with a similarity of 55 %, clustering 44 strains, 59 % of which were isolated from hosts other than soybean. New clusters were also observed for the IGS region. The largest number of differences was detected in the analysis of the 23S rRNA gene, and 16 groups and isolated strains were joined at a very low level of similarity (16 %). In a combined analysis with the three ribosomal regions, the majority of strains isolated from soybean clustered with a similarity of 54 % with type and reference strains of B. japonicum, while most strains isolated from Brazilian indigenous legume species grouped with B. elkanii at a level of similarity of 46 %. All strains were clustered at a very low level of similarity (27 %), and at least two new clusters were clearly defined. These new clusters might be related to intraspecific differences or to novel subspecies, or even to novel species; indeed, strains from one of these clusters show higher 16S rRNA gene sequence similarity to members of the genus Burkholderia. The results obtained in this study emphasize the high level of diversity of symbiotic diazotrophic bacteria in the tropics that still remains to be determined. INTRODUCTIONThe Leguminosae (known as the Fabaceae in the USA) is one of the largest families of plants, with over 18 000 species classified into around 650 genera, representing approximately one-twelfth of all known flowering plants and occupying nearly all terrestrial biomes (Polhill & Raven, 1981). Many species within this family are capable of establishing symbioses with a group of bacteria collectively called rhizobia, of which the most important feature is the capacity...

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confidence: 92%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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RFLP analysis of the rRNA operon of a Brazilian collection of bradyrhizobial strains from 33 legume species

Germano

Menna

Mostasso

et al. 2006

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLO

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“…A correlação entre resultados de hibridização DNA:DNA e do perfil eletroforético de proteínas (Vandamme et al, 1997) tem sido evidenciada. Este método permite, principalmente, diferenciar estirpes e espécies (Rademaker & Bruijin, 1997), sendo considerado uma ferramenta útil para o estudo da diversidade de microrganismos em geral e tem sido aplicado no estudo de diazotróficos simbióticos (Moreira et al, 1993;Dupuy, et al, 1994, Lajudie et al, 1994. No entanto, poucos trabalhos encontrados na literatura utilizaram esta técnica para caracterização de bactérias associativas diazotróficas (Santos et al, 2001).…”

Section: Introductionunclassified

Caracterização fenotípica e diversidade de bactérias diazotróficas associativas isoladas de solos em reabilitação após a mineração de bauxita

Nóbrega

Moreira

Siqueira

et al. 2004

Rev. Bras. Ciênc. Solo

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RESUMOAs gramíneas, usadas comumente para reabilitação de áreas degradadas, podem formar associações em suas raízes com bactérias fixadoras de nitrogênio e, assim, contribuir para a sustentabilidade do ecossistema. Por outro lado, a diversidade microbiana exerce papel relevante na resiliência dos processos biológicos do solo, que incluem a fixação biológica de N 2 . O presente estudo teve como objetivo a caracterização fenotípica de 72 isolados de bactérias diazotróficas associativas Gram-negativas, oriundos de áreas sob diferentes estratégias de reabilitação, após a mineração da bauxita em Poços de Caldas (MG), por meio da inoculação de amostras de solo nos meios de cultivo NFb, Fam e JNFb. Estirpes tipo e referência de espécies de Azospirillum, Herbaspirillum e Burkholderia foram usadas para comparação, uma vez que são capazes de crescer nestes meios. O dendrograma de similaridade baseado em sete características culturais dos isolados em meio de cultivo GNA revelou grande diversidade, sendo obtidos 50 grupos a 81 % de similaridade. A tolerância a NaCl em meio de cultivo batata/sacarose/ácido málico variou de 0 a 50 g L -1 , permitindo separar os isolados e estirpes tipo em cinco grupos. O diâmetro celular variou de 0,61 a 1,21 µm, e 13 isolados diferiram das estirpes tipo e referência. Os padrões de proteína total, obtidos por eletroforese em gel de poliacrilamida (SDS-PAGE), foram bastante diversos, possibilitando a formação de 15 grupos com similaridade a 75 %. Não houve relação entre os grupos formados pelas várias características estudadas, tampouco destes com as áreas de origem dos isolados. Os meios de cultivo Fam e JNFb detectaram, além das espécies-alvo, outras não identificadas. A alta dissimilaridade da maioria dos

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“…Por exibirem crescimento rápido em meio YMA com azul de bromotimol (dois a cinco dias) e terem causado reação ácida nos meios líquido e sólido (coloração amarela), os isolados INPA R-926, R-957 e R-975 provavelmente pertençam ao gê-nero Rhizobium 16 . Ao contrário, Bradyrhizobium pode ser o gênero do qual faz parte o INPA R-991, pois, além de apresentar crescimento relativamente lento (cinco a sete dias), elevou o pH do meio líquido e alcalinizou o meio sólido (coloração azul), características fisiológicas comuns para esse táxon 15,21,36 .…”

Section: Resultsunclassified

Produção de amilase por rizóbios, usando farinha de pupunha como substrato

Oliveira¹,

Oliveira²,

Andrade³

et al. 2007

Ciênc. Tecnol. Aliment.

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ResumoAs amilases estão entre as mais importantes enzimas industriais e são de grande interesse na biotecnologia atual. Embora elas possam ser derivadas de diversas fontes, as de origem microbiana são geralmente as mais procuradas pelas indústrias. As espécies do gênero Bacillus são consideradas as principais fontes de amilases. Apesar disso, a busca por novas fontes microbianas vem crescendo em todo o mundo. O presente estudo objetivou avaliar a produção de amilase por rizóbios nativos, usando farinha de pupunha como substrato. Neste estudo, foi adotado o delineamento experimental inteiramente casualizado, com três repetições. Foram determinados ainda os coeficientes de correlação de Pearson entre as variáveis pH do meio, proteína extracelular, biomassa celular, diâmetro médio da colônia (DMC), diâmetro médio do halo (DMH), índice enzimático (IE) e atividade amilolítica das bactérias selecionadas. Dos 19 isolados com atividade amilolítica em meio YMA modificado, sete (36,8%) exibiram "IE" ≥ 2,1, o que permite considerá-los bons produtores de amilase. Os "IE" apresentados pelos isolados INPA R-987, 950 e 915B foram significativamente inferiores (p < 0,01) aos mostrados pelas bactérias INPA R-926, 975 e 957. A atividade amilolítica variou significativamente (p < 0,01) entre as bactérias investigadas. Os isolados INPA R-975 e R-926 apresentaram, respectivamente, a maior (1,00 U.min -1 .mL -1 ) e a menor (0,31 U.min -1 .mL -1 ) média de atividade. Em termos gerais, a proteína extracelular correlacionou-se positivamente com "IE" (r = 0,52*; p < 0,05) e "DMH" (r = 0,55*; p < 0,05). A biomassa celular apresentou correlações positivas com atividade amilolítica (r = 0,55*; p < 0,05) e "DMH" (r = 0,54*; p < 0,05) e negativa com o pH final do meio de cultivo (r = 0,93**; p < 0,01). Palavras-chave: atividade amilolítica; rizóbios; farinha de pupunha; Amazônia Central. AbstractAmylases are among the most important industrial enzymes and are of great significance in present-day biotechnology. Although they can be derived from various sources, enzymes from microbial sources are generally the most required by industry. Species of the genus Bacillus are considered to be the main sources of amylases, although screening for new microbial sources is increasing all over the world. The objective of the present study was to evaluate the production of amylase by indigenous rhizobia, using peach palm flour as substrate. In this study, a completely randomized experimental design was adopted with three replicates. Pearson's correlation coefficients were calculated for medium pH, extracellular protein, cellular biomass, mean colony diameter (MCD), mean halo diameter (MHD), enzymatic index (EI) and amylolytic activity variables from the bacteria selected. Out of the 19 rhizobia strains with amylolytic activity on modified YMA, seven (36.8%) strains showed "EI" ≥ 2,1, and they were considered as good producers of amylase. The "IE" presented by bacteria INPA R-987, R-950 and R-915B were significantly lower (p < 0.01) than those shown by the ...

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Characterization of Rhizobia Isolated from Different Divergence Groups of Tropical Leguminosae by Comparative Polyacrylamide Gel Electrophoresis of their Total Proteins

Cited by 131 publications

References 36 publications

RFLP analysis of the rRNA operon of a Brazilian collection of bradyrhizobial strains from 33 legume species

RFLP analysis of the rRNA operon of a Brazilian collection of bradyrhizobial strains from 33 legume species

Caracterização fenotípica e diversidade de bactérias diazotróficas associativas isoladas de solos em reabilitação após a mineração de bauxita

Produção de amilase por rizóbios, usando farinha de pupunha como substrato

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