ABSTRACT Termites can create structures that alter the physical and chemical properties of soils. In this process, termites are selective about the soil constituents they will use to construct their mounds. Considering the common occurrence of termite mounds in Brazilian soils, this study aimed to investigate the selective action of termites in the mound building process. Samples were collected from six termite mounds and control soils (at a distance of 15 to 30 m from the termite mound) in different regions in Brazil to analyze the fine earth fraction. The content of clay fraction, organic C and Fe in pedogenic iron oxides increased in the mounds resulting in specific surface area increments. X-ray diffraction indicated a selectivity of termites by clay-sized particles such as kaolinite, gibbsite and iron oxides (hematite and goethite) rather than larger particles such as quartz. The proportion of low-crystalline iron oxides and the maghemite amount decreased in the mounds. The change of color parameters in the termite mounds was due to a combination of increase in clay fraction, organic carbon and iron oxides. The techniques used were sensitive, indicating changes and similarities between the control soils and the termite mounds.

ABSTRACT The two main bottlenecks for a successful phytoremediation program are the metal availability in soil and the metal uptake and transfer to shoots of high biomass plants. Several agronomical practices have been tested to boost the bioavailability of metals in soils and accumulation in plants. Here we assessed the feasibility of plant-growth-promoting bacteria (PGPB) isolated from a site contaminated by gold ore processing activities to assist the phytoremediation of As, Cd, and Pb by Sorghum bicolor and mitigate the metal toxicity in plants. The bacteria Kluyvera intermedia, Klebsiella oxytoca, and Citrobacter murliniae were evaluated in single, double, and triple inoculations. They are regarded as metal resistant and were isolated from the rhizosphere of species naturally growing on the metal contaminated site. The treatments comprised two soils (contaminated and non-contaminated) and single (K. intermedia, K. oxytoca, or C. murliniae) or multiple inoculations (K. intermedia + K. oxytoca; K. intermedia + C. murliniae; K. oxytoca + C. murliniae; K. intermedia + K. oxytoca + C. murliniae). Plants were grown for 42 days after inoculation. The results showed that the PGPB K. oxytoca and the combination of K. intermedia + K. oxytoca and K. intermedia + C. murliniae were able to mitigate the metal toxicity in the contaminated soil and hence increase the shoot biomass, with implications to the effectiveness of phytoextraction. The sorghum ability to translocate Cd to shoots in the contaminated soil was enhanced through the single inoculation with K. oxytoca, C. murliniae, and K. oxytoca, as well as by the joint-inoculation with K. oxytoca + C. murliniae, and K. intermedia + K. oxytoca + C. murliniae. Higher accumulation of metals in shoots is a crucial factor in successful phytoextraction. Arsenic and Pb, on the other hand, had their uptake and concentration in roots stimulated by the inoculation. Therefore, regarding these two metals, phytostabilization programs could benefit from the use of the bacteria studied here.

ABSTRACT There is a complex interaction between various components of the soil ecosystem, including microbial biomass and soil chemical contaminants such as heavy metals and radionuclides, which may greatly affect the efficiency of bioremediation techniques. The aim of this study was to investigate microbial capacity to change pH, changes in the metal soluble-exchangeable fraction, and effects of initial heavy metal contents on soil samples in microbial solubilization/immobilization capacity. The soil samples used in this study were collected at a known metal-contaminated site. Three highly metal-resistant bacteria were isolated from rhizosphere soil samples collected on weed species identified as Senecio brasiliensis, Senecio leptolobus, and Baccharis trimera. A completely randomized experimental design in a factorial arrangement was used, with three replicates. In general, with an acid pH, the isolates neutralized the contaminated growth media. In a neutral or basic initial pH, increases in pH were observed in the media, so these bacteria have an alkalizing effect on the growth media. Soluble metal contents were quite different and depend on the microbial species and heavy metal contents in the soil samples. The soluble-exchangeable fraction of metal such as Cu, Zn, Ni, Cr, Cd, Pb, and Ba may be unavailable after inoculation with heavy metalresistant rhizobacteria. A promising approach seems to be the application of inoculants with metal-resistant bacteria in bioremediation of multi-metal-polluted environments to improve the efficiency of this environmentally friendly technology.

RESUMO: Embora os rizóbios não apresentem a capacidade de fixar nitrogênio em plantas de arroz, estes podem estimular o crescimento da parte aérea e da raiz, aumentando a absorção de nutrientes, como o nitrogênio, e a produtividade. Com o trabalho, avaliou-se o efeito de rizóbios em promover o crescimento de plantas de arroz em função de doses de nitrogênio (N). Efetuou-se um experimento com vasos contendo solo e plantas da cultivar IRGA 424. As plantas foram inoculadas com oito isolados de rizóbios e adubadas com doses equivalentes a 0, 40, 80 e 160kg N ha-1. Avaliou-se a matéria seca e a quantidade de N acumulada na parte aérea e o perfilhamento das plantas. O efeito do N no crescimento de arroz aumentou com a inoculação de rizóbios. A matéria seca da parte aérea das plantas inoculadas e que receberam, aproximadamente, metade da maior dose de N foi equivalente às que receberam a maior dose desse nutriente e sem inoculação. Além disso, a inoculação com rizóbios aumentou o perfilhamento e a absorção de N. A inoculação de rizóbios em arroz, embora não dispense a adubação nitrogenada, tem potencial para aumentar a produção e a eficiência do uso do N.

ABSTRACT: Although rhizobia do not show the ability to fix nitrogen in rice plants, they can stimulate shoot and root growth and even increase productivity and also can increase the plant efficiency in the absorption of nutrients such as nitrogen. This study evaluated the effect of rhizobia in promoting the growth of rice plants fertilized with nitrogen (N). An experiment was developed in pots containing soil and rice of the cultivar IRGA 424. The plants were inoculated with eight isolates of rhizobia and fertilized with 0, 40, 80 and 160kg N ha-1. We evaluated the dry matter of shoots, tillering and amounts of N absorbed by the shoots. The effect of N on rice growth increased by inoculation of rhizobia. Inoculated plants and fertilized with approximately half of the higher N dose, produced shoot equivalent of the plants that received the highest dose of this nutrient and without inoculation. Moreover, inoculation with rhizobia increased tillering and absorption of N. The practice of rhizobia inoculation in rice, although not replace the nitrogen fertilization, have great potential for rice farmers, allowing greater production and efficiency of N use.

ABSTRACT Rhizobia and associative bacteria promote growth in rice plants (Oryza sativa L.) through a series of mechanisms, but most studies on inoculation have been performed based on inoculation with these bacteria in a separate or singular manner. The objective of this study was to assess the efficiency of single/isolated inoculation and inoculation combined with symbiotic rhizobia from forage legume and with Azospirillum brasilense on promoting growth and the root colonization process in wetland rice. Two rhizobia among four isolates from a greenhouse and a laboratory experiment were selected that efficiently promoted seed germination and rice plant growth in a sterilized substrate and in soil. The two most efficient isolates (UFRGS Vp16 and UFRGS Lc348) were inoculated alone or in combination with a commercial product containing A. brasilense in two field experiments using two wetland rice cultivars over two growing seasons. In the field experiments, these isolates coinoculated with A. brasilense promoted larger increases in the agronomic variables of wetland rice compared to the control without inoculation. Confocal laser microscopy confirmed the presence of inoculated bacteria tagged with gfp (UFRGS Vp16, UFRGS Lc348, and A. brasilense) colonizing the root surface of the rice seedlings, mainly in the root hairs and lateral roots.

This work aimed to evaluate the symbiotic compatibility and nodulation efficiency of rhizobia isolated from Desmodium incanum, Lotus corniculatus, L. subbiflorus, L. uliginosus and L. glaber plants by cross-inoculation. Twelve reference strains and 21 native isolates of rhizobia were genetically analyzed by the BOX-PCR technique, which showed a high genetic diversity among the rhizobia studied. The isolates were also characterized based on their production of indolic compounds and siderophores, as well as on their tolerance to salinity. Fifteen of the 33 rhizobia analyzed were able to produce indolic compounds, whereas 13 produced siderophores. All the tested rhizobia were sensitive to high salinity, although some were able to grow in solutions of up to 2% NaCl. Most of the native rhizobia isolated from L. uliginosus were able to induce nodulation in all plant species studied. In a greenhouse experiment using both D. incanum and L. corniculatus plants, the rhizobia isolate UFRGS Lu2 promoted the greatest plant growth. The results demonstrate that there are native rhizobia in the soils of southern Brazil that have low host specificity and are able to induce nodulation and form active nodules in several plant species.

O cornichão é uma leguminosa forrageira perene hiberno-primaveril de grande importância para o Rio Grande do Sul, destacando-se pela capacidade de se manter em solos relativamente ácidos e pouco férteis. Com este trabalho, objetivou-se a seleção de rizóbios para cornichão tolerantes à acidez e ao Al tóxico e eficientes na fixação biológica de N em solos de baixa fertilidade. Foram avaliados 52 isolados de rizóbios de Lotus spp. obtidos de solos de cinco localidades do Rio Grande do Sul. Os rizóbios foram avaliados quanto à diversidade genética, à tolerância a pH 4,2 e ao Al tóxico. Entre os rizóbios tolerantes a fatores de acidez, sete foram avaliados quanto à eficiência simbiótica com plantas, em casa de vegetação, em vasos com solo não estéril. Observou-se alta diversidade genética entre os rizóbios estudados, dos quais 16 foram tolerantes a pH 4,2 e a 50 µM de Al em meio de cultura, produzindo populações da ordem de 10(7) até 10(8) UFC mL-1. Os sete rizóbios testados em casa de vegetação superaram as estirpes atualmente recomendadas para a produção de inoculantes, o que demonstra a existência, em solos do Rio Grande do Sul, de rizóbios tolerantes à acidez do solo e eficientes como fixadores de N em plantas de cornichão.

Birdsfoot trefoil is a perennial winter-spring forage legume, highly important in Rio Grande do Sul, in the south of Brazil. It stands out for the ability to survive in relatively acidic and nutrient-poor soils. This study aimed to select birdsfoot trefoil rhizobial strains with tolerance to acidity and aluminum toxicity and efficiency in nitrogen fixation in nutrient-poor soils. Fifty-two isolates of Lotus spp rhizobia from soils of five locations in the state of Rio Grande do Sul were evaluated. The strains were evaluated for genetic diversity and tolerance to pH 4.2 and to aluminum toxicity. Of the acid-tolerant rhizobia, seven were evaluated for symbiotic efficiency in plants grown in pots with non-sterile soil, in a greenhouse experiment. Genetic diversity in the studied rhizobia was high and 16 were tolerant to pH 4.2 and to 50 µM Al in media broth, reaching populations from 10(7) up to 10(8) CFU mL-1. The seven rhizobia tested in the greenhouse performed better than the strains recommended for inoculant production, demonstrating the existence of rhizobia tolerant to soil acidity and efficient in nitrogen fixation on birdsfoot trefoil plants in soils of Rio Grande do Sul.

A inoculação de plantas leguminosas com rizóbios é um dos principais métodos biotecnológicos de utilização de micro-organismos em plantas visando à fixação biológica de nitrogênio na agricultura. No entanto, nos últimos anos, vêm sendo observada nesses micro-organismos a capacidade de produção de fitohormônios, principalmente o ácido indol-acético (AIA) e a promoção de crescimento em gramíneas. Dessa forma, os objetivos deste trabalho foram quantificar o ácido indol-acético produzido por rizóbios isolados de alfafa, avaliar o efeito da inoculação desses micro-organismos na germinação de sementes de arroz e realizar a caracterização genética desses isolados. Nove rizóbios isolados de nódulos de alfafa foram avaliados quanto a sua capacidade de produção de equivalentes de AIA e a influência da inoculação desses micro-organismos na germinação e desenvolvimento de plântulas de arroz. Os rizóbios produtores de AIA foram identificados pelo sequenciamento da região do gene 16S do DNAr. A produção de equivalentes ao ácido indol-acético foi observada em todos rizóbios, com valores que variaram de 43,04 a 101,26µg mL-1 em meio de cultura. Com relação à germinação das sementes de arroz, a inoculação com rizóbios acelerou o processo e o crescimento de suas plântulas. Os rizóbios UFRGS Ms58, Ms515, Ms195, Ms205, Ms2010 e 2012 foram identificados como pertencentes à espécie Sinorhizobium meliloti e UFRGS Ms55, Ms72 e Ms75 à espécie Rhizobium sp.

The inoculation of leguminous plants with rhizobia is one of the main methods of biotechnological use of microorganisms in order to obtain biological nitrogen fixation in agriculture. However, in recent years it has been attributed to these microorganisms the ability to produce phytohormones, mainly indole acetic acid (IAA), and to promote the growth in grasses. Thus, the objectives of this study were to quantify the indole acetic acid produced by rhizobia from alfalfa and to evaluate the effect of inoculation of these microorganisms on the germination of rice seed and to perform the genetic characterization of these isolates. Nine rhizobia, from nodules of alfalfa, were evaluated for their ability to produce IAA equivalents and for their influence in inoculating these microorganisms on germination and seedling development of rice. Moreover, these rhizobia producers of IAA were identified by the 16S region of DNAr. The equivalent production of indole acetic acid was observed in all tested isolates, with values ranging from 43.04 to 101.26µg mL-1 in culture medium. Regarding the germination of rice seeds, the inoculation with rhizobia accelerated this germination and its growth. Microorganisms UFRGS Ms58, UFRGS Ms515, UFRGS Ms195, UFRGS Ms205, UFRGS Ms2010 and UFRGS 2012 were identified as belonging to the species of Sinorhizobium meliloti. Microorganisms Ms55 UFRGS, UFRGS Ms75 and UFRG Ms72 were identified as belonging to the species of Rhizobium sp.

O cornichão (Lotus corniculatus L.) tem sido apontado como uma das forrageiras mais promissoras para o uso no Rio Grande do Sul. No entanto, pouco se conhece a respeito da interação dessas plantas com os rizóbios nativos em condições de campo. Este estudo visou a avaliar a eficiência das interações entre rizóbios e plantas de Lotus corniculatus cv. 'São Gabriel' em condições de campo. O experimento foi conduzido na Estação Experimental Agronômica da UFRGS com delineamento de blocos ao acaso com quatro repetições. Os tratamentos estudados foram: dois controles sem inoculação (um com a adição de 80kg ha-1 de nitrogênio na forma de uréia e outro sem a adição de nitrogênio); inoculação com rizóbios: estirpes SEMIA 816, recomendada no Brasil, U510, recomendada no Uruguai, U512, em estudo no Uruguai, e quatro isolados de rizóbios nativos (UFRGS Lc322, UFRGS Lc349, UFRGS Lc269 e Iso 7). O isolado UFRGS Lc322 e a estirpe U510 são mais eficientes do que a estirpe SEMIA 816, recomendada para cornichão no país. Além disso, a produção de forragem das plantas inoculadas com o isolado UFRGS Lc322 e a estirpe U510 superou a obtida com a adição de 80kg ha-1 de nitrogênio.

The bird's-foot trefoil (Lotus corniculatus L.) has been pointed as one of the most promising forage plants for use in Rio Grande do Sul. However, there is lack of information about the interaction of those plants with the native rhizobia in field conditions. This study aimed to evaluate the efficiency of interactions between rhizobia and plants of Lotus corniculatus cv. 'São Gabriel' in field conditions. The experiment was carried out in the Agronomic Experimental Station (EEA) of the Federal University of Rio Grande do Sul (UFRGS), using random block design with four replications. The treatments were: two controls without inoculation (one with the addition of urea corresponding to 80kg ha-1 of nitrogen and other without nitrogen addition); inoculation with rhizobia: strain SEMIA 816, recommended in Brazil, U150 recommended in Uruguay and U512, being studied in Uruguay, and four rhizobia isolates (UFRGS Lc322, UFRGS Lc349, UFRGS Lc269 e Iso 7). The rhizobia isolate Lc322 and the strain U510 are more efficient than the strain SEMIA 816, which is recommended for bird's-foot trefoil in the country. Furthermore, the forage production of plants inoculated with the isolate Lc322 and the strain U510 surpassed that, with the addition of 80kg ha-1 of nitrogen.

As leguminosas forrageiras, além de contribuírem para o aumento da oferta de forragem com alto teor de proteína, também são importantes como cobertura vegetal de solos. Entre as leguminosas com grande potencial forrageiro, espécies do gênero Lotus têm se destacado. Este trabalho visou à seleção de rizóbios nativos eficientes em Lotus glaber a partir de amostras de solo de cinco localidades do Rio Grande do Sul. Obtiveram-se 259 isolados de rizóbios, que foram avaliados quanto à morfologia colonial e produção de melanina. Destes, 15 isolados foram selecionados para avaliação da eficiência na fixação simbiótica de nitrogênio em experimento em casa de vegetação e caracterizados geneticamente por comparação do perfil eletroforético dos produtos de amplificação do DNA genômico, por PCR com os oligonucleotídeos iniciadores BOX e ERIC. Apenas cinco isolados produziram melanina. Nove isolados foram mais eficientes do que a estirpe SEMIA 830, que é autorizada para a produção de inoculante para L. glaber no país. Na caracterização genética, observou-se que nenhum isolado apresentou identidade com as estirpes recomendadas, o que demonstra que, nos solos do Rio Grande do Sul, existem rizóbios autóctones eficientes na fixação simbiótica de nitrogênio com L. glaber, podendo ser recomendados para estudos a campo, visando a uma futura produção de inoculantes para estas leguminosas no Brasil.

Forage legumes besides contributing to increase the supply of high protein content forage are also good cover for the soil surface. Among the legumes with high potential forage, the genus Lotus has been outstanding. This research aimed to selectnative rhizobia efficient in fixing nitrogen with Lotus glaber, from soil samples of five localities of Rio Grande do Sul. A total of 259 rhizobia isolates were obtained, which were evaluated based on colony morphology and in vitro melanin production. Among them, 15 isolates were selected for the evaluation of the symbiotic nitrogen fixation efficiency in greenhouse experiments and genetically characterized by genomic DNA fingerprinting, PCR amplification with BOX and ERIC primers. Only five isolates produced melanin. Nine isolates were more efficient than strain SEMIA 830, which is authorized for the production of commercial inoculants of L. glaber in Brazil. In relation to the genetic characterization, the isolates have shown no similarity with the commercial strains, indicating that in the soils of Rio Grande do Sul there are autochthonous rhizobia capable of efficient symbiotic nitrogen fixation with L. glaber, which might be recommended for field evaluations, aiming a future production of inoculants for this legume in Brazil.

O presente trabalho teve como objetivos avaliar a quantidade de nitrogênio imobilizado e a sua remineralização, nos sistemas plantio direto (SPD) e convencional (SC), ao longo do ciclo do milho. Para tal, foram coletadas amostras da camada de 0-5cm de solo semeado com milho em sucessão à aveia preta, sob os sistemas convencional e plantio direto, no dia da semeadura do milho e 46, 62, 88 e 112 dias após. Amostras da parte aérea das plantas de milho foram coletadas nas mesmas épocas, com exceção da primeira, e avaliadas quanto à quantidade de nitrogênio acumulado. As amostras de solo foram avaliadas quanto à quantidade de nitrogênio mineral no solo, de nitrogênio e carbono potencialmente mineralizáveis, atividade de urease e de nitrogênio imobilizado na biomassa microbiana. A imobilização microbiana do nitrogênio foi maior no sistema plantio direto, levando a uma menor quantidade de nitrogênio mineral no solo e resultando em menor acúmulo de nitrogênio na parte aérea do milho ao final do seu ciclo neste sistema, em comparação com o convencional. Não foi observada remineralização do nitrogênio imobilizado, indicando que a biomassa microbiana atuou mais como agente da mineralização de nitrogênio orgânico do que como fonte de nitrogênio potencialmente mineralizável.

The present research aimed to evaluate the amount of nitrogen immobilized and its remineralization, in conventional and in no-till systems, during the corn cycle. A soil that was used for oats crop was seeded with corn and subjected to under conventional and no-till systems. Samples were collected from the top (0-5cm) soil layer on the day of the corn seeding and after 46, 62, 88 and 112 days. At the same time, the aerial parts of corn plants were collected and nitrogen accumulation was evaluated. Soil samples were assayed for mineral N, potentially mineralizable N and C, activity and immobilized N. Microbial nitrogen immobilization was higher in no-till system, reducing the soil mineral nitrogen concentration and resulting in a lower amount of nitrogen in the aerial parts of corn plants at the end of the growth cycle. Nitrogen remineralization was not observed, indicating that the microbial biomass acted more as an agent of organic N mineralization than as a source of potentially mineralizable N.

Variantes em estirpes de rizóbio usadas na produção de inoculantes não são desejáveis e podem propiciar resultados inesperados na inoculação da leguminosa. Estudou-se a variabilidade nas características fenotípicas e a estabilidade genética das estirpes de Bradyrizobium recomendadas para a inoculação em soja. Foram isolados variantes com morfologia colonial estável das estirpes SEMIA 5080 de Bradyrhizobium japonicum e SEMIA 5019 de B. elkanii. Variantes da estirpe SEMIA 587, obtidas por outro autor, também foram estudadas. As variantes diferiram nas características coloniais, capacidade de nodulação, eficiência na fixação de nitrogênio, e competitividade para formação de nódulos em duas variedades de soja (Jacui 7 e IAS 5). O comportamento simbiótico das variantes diferiu de acordo com as variedades onde foram inoculadas. Somente as variantes 5019 G e 5019 P, originadas da estirpe SEMIA 5019, diferiram quanto ao perfil isoenzimático. Observou-se diferenças na resistência a antibióticos entre as variantes das duas estirpes. Análises relacionando as características simbióticas com morfologia colonial ou resistência a antibióticos não foram conclusivas. A variabilidade em estirpes de rizóbio pode ser um fator importante a ser considerado nos programas de seleção de estirpes e no monitoramento da preservação das culturas bacterianas para a produção de inoculantes.

Variation in rhizobia strains isn’t a desirable fact based mainly on the possibility of unexpected results on legume inoculation. In this work, we studied the variability on phenotypic characteristics and genetic stability of rhizobia strains recommended for soybean inoculation. Variants with stable colony morphology were obtained from Bradyrhizobium japonicum strain SEMIA 5080 and from B. elkanii SEMIA 5019. Variants from SEMIA 587 obtained by another author were also used. The variants differed on colony characteristics, nodulation capacity, nitrogen fixation efficiency and competitive ability for nodule formation in two soybean varieties (Jacui 7 and IAS 5). Symbiotic behavior varied according to plant variety. Only the variants 5019 G and 5019 P differed on the isoenzymatic profile. There were differences in antibiotic resistance between variants from two strains. Correlation between symbiotic characteristics and colony morphology or antibiotic resistance wasn’t conclusive. The results indicate that the variability in rhizobia strains might be an important factor to be considered in strain selection and preservation of cultures for inoculant production.

Este artigo visa contribuir para a discussão relacionada ao futuro da microbiologia no Brasil. Este trabalho apresenta um levantamento, realizado em 2001, que mostra um quadro da situação atual da microbiologia no Brasil. A maioria das conclusões são fundamentadas nos dados coletados dos cursos universitários e dos dados fornecidos pelo Conselho Nacional de Pesquisa (CNPq) brasileiro e pela Coordenação para o Aperfeiçoamento de Pessoal de Nível Superior (CAPES). No entanto, algumas das afirmativas são baseadas mais nas experiências de ensino de pós-graduação em microbiologia do que nos dados fornecidos pelas instituições oficiais. Tais afirmativas foram feitas no sentidos de incrementar o desenvolvimento da microbiologia no Brasil. Além disso, é apresentado um plano que propõe maior número de bolsas concedidas para a área de microbiologia, para aumentar a possibilidade de formar uma massa crítica necessária para maior desenvolvimento da microbiologia brasileira, visando produzir discussão e mais debate sobre o futuro da microbiologia no Brasil.

This paper aims to contribute to the discussion and concern on the future of microbiology in Brazil. This work presents a survey carried out in 2001 that shows a picture of Microbiology status in Brazil. Most of the conclusions are based on the data collected from the university courses and from the data supplied by the Brazilian National Research Council (CNPq) and the Coordination for the Improvement of Higher Level Personnel (CAPES). However, some of the affirmatives are more based on the experiences with postgraduate teaching in microbiology than on the data provided by official institutions. Such affirmatives were made in order to improve the development of the microbiology in Brazil. Also, a plan that proposes higher number of scholarships awarded to microbiology, to increase the possibility to form a critical mass necessary for further development of the Brazilian microbiology, is presented in order to produce discussion and more debate on the future of the Microbiology in Brazil.