Effects of rumen microorganisms on the decomposition of recycled straw residue / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Recently, returning straw to the fields has been proved as a direct and effective method to tackle soil nutrient loss and agricultural pollution. Meanwhile, the slow decomposition of straw may harm the growth of the next crop. This study aimed to determine the effects of rumen microorganisms (RMs) on straw decomposition, bacterial microbial community structure, soil properties, and soil enzyme activity. The results showed that RMs significantly enhanced the degradation rate of straw in the soil, reaching 39.52%, which was 41.37% higher than that of the control on the 30th day after straw return. After 30 d, straw degradation showed a significant slower trend in both the control and the experimental groups. According to the soil physicochemical parameters, the application of rumen fluid expedited soil matter transformation and nutrient buildup, and increased the urease, sucrase, and cellulase activity by 10%‒20%. The qualitative analysis of straw showed that the hydroxyl functional group structure of cellulose in straw was greatly damaged after the application of rumen fluid. The analysis of soil microbial community structure revealed that the addition of rumen fluid led to the proliferation of Actinobacteria with strong cellulose degradation ability, which was the main reason for the accelerated straw decomposition. Our study highlights that returning rice straw to the fields with rumen fluid inoculation can be used as an effective measure to enhance the biological value of recycled rice straw, proposing a viable solution to the problem of sluggish straw decomposition.

Efecto de inoculantes y aditivos sobre fracciones de fermentación ruminal y degradación in vitro en ensilaje de sorgo (Sorghum sp) / Effect of inoculants and additives on fractions of ruminal fermentation and in vitro degradation in sorghum silage (Sorghum sp)

Resumen La acción de inoculantes y aditivos sobre la composición química y degradación ruminal de la materia seca (MS) de ensilaje de sorgo ya ha sido reportada. Sin embargo, se desconoce el efecto que estos tienen sobre la actividad microbiana, y por lo tanto, sobre su potencial de asimilación a nivel ruminal. El objetivo de este estudio fue evaluar el efecto de la adición de inoculantes y aditivos en ensilaje de la planta completa de Sorghum sp. sobre las fracciones de fermentación ruminal y degradación in vitro. Se utilizó un diseño experimental completamente al azar, con 7 tratamientos (control, 3 inoculantes, 2 aditivos y una combinación de inoculante con aditivo) y 21 repeticiones por tratamiento. Se usaron diferentes tipos de inoculantes y aditivos en el proceso de ensilaje de sorgo: lactosuero, yogur, ácido fosfórico, Lactobacillus plantarum y mezcla mineral. Después de 60 d de fermentación, se cuantificaron MS, pH, proteína cruda (PC), degradación in vitro de la MS a 24 h (DIVMS), parámetros de la cinética de fermentación (Vmáx = volumen máximo, L = fase lag, S = tasa de fermentación) y volumen fraccional (fermentación rápida = 0 h a 8 h, media = 8 h a 24 h y lenta = 24 h a 72 h) por producción de gas in vitro. El análisis estadístico indicó que los inoculantes y aditivos modificaron la composición química, los parámetros L, Vmáx, DIVMS y las fracciones de fermentación rápida y media (P < 0.05). El uso de lactosuero y L. plantarum como inoculantes en ensilaje de sorgo redujo la pérdida de PC y, en general, los inoculantes y aditivos mejoraron el pH de los ensilajes; no obstante, disminuyeron el aprovechamiento de las fracciones de fermentación rápida y media, modificaron el potencial de fermentación y provocaron efecto negativo en la DIVMS.

Abstract The action of inoculants and additives on the chemical composition and ruminal degradation of sorghum silage dry matter (DM) has already been reported. However, the effect that these have on the microbial activity and, therefore, on its potential for assimilation at the ruminal level is unknown. For that reason, the objective of this study was to evaluate the effect of the addition of inoculants and additisves in silage of the entire plant of Sorghum sp. on fractions of ruminal fermentation and in vitro degradation. The experimental design was completely random, with 7 treatments (3 inoculants, 2 additives and a combination of inoculate with additive) and 21 repetitions per treatment. Different types of inoculants and additives were used in the process of sorghum silage: cheese whey, yogurt, phosphoric acid, Lactobacillus plantarum and mineral mixture. After 60 d of fermentation, dry matter (DM), hydrogen potential (Hp), crude protein (CP), in vitro dry matter degradation to 24 h (IVDMD), parameters of fermentation kinetics (Vmax= maximum volume L = lag phase, S = fermentation rate) and fractional volume (rapid fermentation = 0 h to 8 h, intermediate = 8 h to 24 h and slow = 24 h to 72 h) were quantified through the in vitro gas production technique. The results indicated that the inoculants and additives modified the chemical composition, the L parameters, Vmax, IVDMD and the rapid and intermediate fermentation fractions (P < 0.05). The use of cheese whey and L. plantarum as inoculants in sorghum silage reduced the loss of CP. Overall, the use of inoculants and additives improved silage Hp; however, it decreased the use of rapid and intermediate fermentation fractions, modified the fermentation potential of the silage and provoked a negative effect on the IVDMD.

Yield increase of corn inoculated with a commercial arbuscular mycorrhizal inoculant in Brazil / Rendimento de grãos da cultura do milho com inoculante micorrízico arbuscular comercial no Brasil

ABSTRACT: Arbuscular mycorrhizal fungi (AMF) play an important role in plant growth. However, there are no reports of legally commercialized AMF-based inoculants for agricultural crops in Brazil. The objective of this research was to evaluate the agronomic efficiency of a Rhizophagus intraradices inoculant in combination with phosphate fertilization in grain yield of corn under different edaphoclimatic conditions in Brazil. Experiments were conducted in five Brazilian states (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul and Santa Catarina) in a 2 x 3 factorial scheme, with two inoculation treatments (inoculated and non-inoculated seeds) and three doses of phosphate fertilization (0, 50 and 100% of the recommended P). At the end of the crop cycle (stages R4-R5), inoculation provided increases in biomass (average of 48%) regardless of the applied dose of P, higher P absorption, and 54% average increase in grain yield. In conclusion, the mycorrhizal inoculant increases biomass yield, P uptake and corn grain yield under different edaphoclimatic conditions in Brazil, especially in soils that originally had low or medium levels of available P.

RESUMO: Os fungos micorrízicos arbusculares (FMA) desempenham um papel importante no crescimento das plantas, no entanto, não há relatos de inoculantes legais baseados em AMF para culturas agrícolas no Brasil. O objetivo deste trabalho foi avaliar a eficiência agronômica de um inoculante a base de Rhizophagus intraradices em combinação com a adubação fosfatada no rendimento de grãos de milho sob diferentes condições edafoclimáticas no Brasil. Os experimentos foram conduzidos em cinco estados brasileiros (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul e Santa Catarina) em esquema fatorial 2 x 3, com dois tratamentos de inoculação (sementes inoculadas e não inoculadas) e três doses de adubação fosfatada (0, 50 e 100% do P recomendado). No final do ciclo da cultura (estágios R4-R5), a inoculação proporcionou aumentos na biomassa (média de 48%), independentemente da dose aplicada de P, maior absorção de P e aumento médio de 54% no rendimento de grãos. Conclui-se que o inoculante micorrízico arbuscular aumenta a produção de biomassa, a absorção de P e a produtividade de grãos de milho em diferentes condições edafoclimáticas no Brasil, especialmente em solos que originalmente apresentavam níveis baixos ou médios de P.

Rhizobacterial inoculants: The formulation as biofertilizer and its application on chili plants (Capsicum annum L.)

Aims@#The use of rhizobacteri as biofertilizer may help plants in obtaining nutrients from soil. A consortium inoculant (co-inoculant) consisting of nitrogen fixing and phosphate solubilizing bacteria is formulated to maintain its ability as booster of plant growth. This is easy to be stored and applied on plants. The aims of the study were to formulate rhizobacterial co-inoculant and its application on chili plants at greenhouse experiment. @*Methodology and results@#Isolates of Burkholderia cepacia KD 2.10, Serratia marcescens KAHN 15.12, and Bacillus thuringiensis SAHA 12.12 which have the ability in fixing nitrogen and solubilizing phosphate were used in this study. The three isolates did not show antagonistic activity and hypersensitivity reaction on chili plant. Biofertilizer as carrier material with talc-based powder was mixed with three isolates. This 109 CFU/g cell population of rhizobacterial consortium could be maintained up to six months of storage. Based on result of completely randomized design (CRD) using two factorials and four replicates, application of rhizobacterial co-inoculant significantly affected plant height, number of leaves, flowering age, dry weight of upper plant and root, and root length of chili plant. @*Conclusion, significance and impact of study@#Rhizobacterial co-inoculant was effective as biofertilizer to improve the growth of chili plants and it reduced the use of chemical fertilizer.

Phenotypic, genetic and symbiotic characterization of Erythrina velutina rhizobia from Caatinga dry forest

Abstract Erythrina velutina ("mulungu") is a legume tree from Caatinga that associates with rhizobia but the diversity and symbiotic ability of "mulungu" rhizobia are poorly understood. The aim of this study was to characterize "mulungu" rhizobia from Caatinga. Bacteria were obteined from Serra Talhada and Caruaru in Caatinga under natural regeneration. The bacteria were evaluated to the amplification of nifH and nodC and to metabolic characteristics. Ten selected bacteria identified by 16S rRNA sequences. They were tested in vitro to NaCl and temperature tolerance, auxin production and calcium phosphate solubilization. The symbiotic ability were assessed in an greenhouse experiment. A total of 32 bacteria were obtained and 17 amplified both symbiotic genes. The bacteria showed a high variable metabolic profile. Bradyrhizobium (6), Rhizobium (3) and Paraburkholderia (1) were identified, differing from their geographic origin. The isolates grew up to 45 °C to 0.51 mol L-1 of NaCl. Bacteria which produced more auxin in the medium with l-tryptophan and two Rhizobium and one Bradyrhizobium were phosphate solubilizers. All bacteria nodulated and ESA 90 (Rhizobium sp.) plus ESA 96 (Paraburkholderia sp.) were more efficient symbiotically. Diverse and efficient rhizobia inhabit the soils of Caatinga dry forests, with the bacterial differentiation by the sampling sites.

Efecto de la inoculación y peletización en la germinación y crecimiento de plantas de maca (Lepidium meyenii W.) a nivel in vitro e invernadero / Effect of inoculation and pelleting on the germination and growth of maca (Lepidium meyenii W.) in vitro and greenhouse

Se compararon semillas inoculadas sin peletizar y peletizadas con tres distintas cepas (C32, DZ50 y AC7) en maca (Lepidium meyenii) a nivel in-vitro e invernadero en donde a nivel in-vitro se midió el porcentaje de germinación y peso seco y a nivel de invernadero el porcentaje de emergencia, peso seco de raíz y peso seco de la parte aérea, encontrándose que tanto en la germinación como en la emergencia los porcentajes fueron mayores en los tratamientos peletizados que los no peletizados. En los pesos secos a nivel in vitro todos los tratamientos peletizados superaron a los tratamientos sin peletizar y los tratamientos con inoculante peletizados y sin peletizar fueron superiores a los controles sin inoculante. Por otro lado se midió la cantidad de células viables y la actividad de agua (Aw) de las semillas peletizadas y sin peletizar en donde se observó que todos los tratamientos peletizados tienden a conservar estos dos parámetros a través del tiempo.

Non-pelleted and pelleted seeds of maca (Lepidium meyenii W.) inoculated with three strains (C32, DZ50 and AC7) were compared at in-vitro and greenhouse tests. Percentage of germination and dry weight of plant were measured in-vitro level. Percentage of emergence, root and shoot dry weight were measured in greenhouse. Both germination and emergence percentages were higher in pelleted than non-pelleted treatments. In vitro dry weights of all the pelleted treatments were higher than non-pelleted; pelleted and non-pelleted inoculated dry weights were higher than controls without inoculant. The amount of viable cells and water activity (Aw) of pelleted and non-pelleted seeds were measured; pelleted treatments tend to preserve these two parameters over time.

Initial growth in maize in compliance of Azospirillum brasilense inoculation and nitrogen rates / Crescimento inicial do milho em função da inoculação de Azospirillum brasilense e doses de nitrogênio

Investigate the effects of the interaction between nitrogen fertilization and inoculation with diazotrophic bacteria becomes important, especially for maize, highly exacting in N. Thus, the aim of this study was to evaluate the initial growth of maize in compliance A. brasilense inoculation and nitrogen rates. The experiment was conducted in a greenhouse, in Mato Grosso do Sul State University, Aquidauana University Unit, between the months of march and April of 2014. It was used completely randomized design in a 2x4 factorial arrangement with four replications. The treatments consisted of the absence and presence of inoculation with A. brasilense and four levels of nitrogen fertilization (0, 25, 50, 100 kg ha-1 N). At 40 days after sowing the following variables were evaluated: leaf area, plant height, stem diameter, dry mass of shoot and of root. There was interaction between doses of nitrogen and inoculation with A. brasilense for most of the evaluated variables. Inoculation with A. brasilense provides improvements in the establishment and initial development of maize. In the presence of inoculation with A. brasilense, the development of maize increases linearly the administration of the nitrogen doses.

Investigar os efeitos da interação entre adubação nitrogenada e inoculação com bactérias diazotróficas tornou-se importante, sobretudo para a cultura do milho, altamente exigente em N. Assim, o objetivo deste trabalho foi verificar o crescimento inicial do milho em função de inoculação A. brasilense e doses de nitrogênio. O experimento foi desenvolvido em estufa agrícola, na Universidade Estadual de Mato Grosso do Sul, Unidade Universitária de Aquidauana, entre os meses de março a abril de 2014. Utilizou-se delineamento experimental inteiramente casualizado em esquema fatorial 2x4, com quatro repetições. Os tratamentos constaram da combinação da ausência e presença de inoculação com A. brasilense e quatro níveis de adubação nitrogenada (0; 25; 50; 100 kg ha-1 de N). Aos 40 dias após a semeadura foram avaliadas as seguintes variáveis: área foliar, altura de plantas, diâmetro do colmo, massa seca da parte aérea e massa seca das raízes. Houve interação entre doses de nitrogênio e inoculação com A. brasilense para a maioria das variáveis avaliadas. A inoculação com A. brasilense proporciona melhorias no estabelecimento e no desenvolvimento inicial do milho. Na presença de inoculação com A. brasilense, o desenvolvimento de milho aumenta linearmente a função das doses de nitrogênio.

Changes in the microbial community during bioremediation of gasoline-contaminated soil

Abstract We aimed to verify the changes in the microbial community during bioremediation of gasoline-contaminated soil. Microbial inoculants were produced from successive additions of gasoline to municipal solid waste compost (MSWC) previously fertilized with nitrogen-phosphorous. To obtain Inoculant A, fertilized MSWC was amended with gasoline every 3 days during 18 days. Inoculant B received the same application, but at every 6 days. Inoculant C included MSWC fertilized with N–P, but no gasoline. The inoculants were applied to gasoline-contaminated soil at 10, 30, or 50 g/kg. Mineralization of gasoline hydrocarbons in soil was evaluated by respirometric analysis. The viability of the inoculants was evaluated after 103 days of storage under refrigeration or room temperature. The relative proportions of microbial groups in the inoculants and soil were evaluated by FAME. The dose of 50 g/kg of inoculants A and B led to the largest CO2 emission from soil. CO2 emissions in treatments with inoculant C were inversely proportional to the dose of inoculant. Heterotrophic bacterial counts were greater in soil treated with inoculants A and B. The application of inoculants decreased the proportion of actinobacteria and increased of Gram-negative bacteria. Decline in the density of heterotrophic bacteria in inoculants occurred after storage. This reduction was bigger in inoculants stored at room temperature. The application of stored inoculants in gasoline-contaminated soil resulted in a CO2 emission twice bigger than that observed in uninoculated soil. We concluded that MSWC is an effective material for the production of microbial inoculants for the bioremediation of gasoline-contaminated soil.

Screening of plant growth promoting bacteria associated with barley plants (Hordeum vulgare L.) cultivated in South Brazil / Seleção de bactérias promotoras de crescimento vegetal associadas com plantas de cevada (Hordeum vulgareL.) cultivadas no Sul do Brasil

The occurrence of associations between bacteria and plant roots may be beneficial, neutral or detrimental. Plant growth promoting (PGP) bacteria form a heterogeneous group of beneficial microorganisms that can be found in the rhizosphere, the root surfaces or in association with host plant. The aim of this study was to isolate and characterize PGP bacteria associated to barley plants (Hordeum vulgare L.) aiming a future application as agricultural inoculant. One hundred and sixty bacterial strains were isolated from roots or rhizospheric soil of barley based on their growth in nitrogen-free selective media. They were evaluated for their ability to produce indolic compounds (ICs) and siderophores, and to solubilize tricalcium phosphate inin vitro assays. Most of them (74%) were able to synthesize ICs in the presence of the precursor L-tryptophan, while 57% of the isolates produced siderophores in Fe-limited liquid medium, and 17% were able to solubilize tricalcium phosphate. Thirty-two isolates possessing different PGP characteristics were identified by partial sequencing of their 16S rRNA gene. Strains belonging to Cedecea andMicrobacterium genera promoted the growth of barley plants in insoluble phosphate conditions, indicating that these bacteria could be used as bioinoculants contributing to decrease the amount of fertilizers applied in barley crops.

A ocorrência de associações entre bactérias e raízes de plantas pode ser benéfica, neutra ou prejudicial. Bactérias promotoras de crescimento vegetal (BPCV) formam um grupo heterogêneo de micro-organismos benéficos que pode ser encontrado na rizosfera, superfícies de raízes ou em associação com plantas hospedeiras. O objetivo deste estudo foi isolar e caracterizar bactérias promotoras do crescimento vegetal (PCV) associadas a plantas de cevada (Hordeum vulgare L.), visando uma futura aplicação como inoculante agrícola. Cento e sessenta linhagens bacterianas foram isoladas a partir de raízes ou solo rizosférico de cevada com base na sua multiplicação em meios seletivos sem nitrogênio. Todos os isolados foram avaliados quanto è sua capacidade de produzir compostos indólicos (CIs), sideróforos e solubilizar fosfato tricálcio, em ensaios in vitro. A maioria dos isolados (74%) foi capaz de sintetizar CIs na presença do precursor L-triptofano, enquanto que 57% produziram sideróforos em meio líquido com deficiência de Fe e 17% foram capazes de solubilizar fosfato tricálcio. Trinta e dois isolados que apresentaram diferentes características PCV foram identificados pelo sequenciamento parcial do gene 16S rRNA. Linhagens pertencentes aos gêneros Cedecea eMicrobacterium promoveram o crescimento de plantas de cevada em condições de fosfato insolúvel, indicando que estas bactérias podem ser utilizadas como inoculantes, contribuindo para a redução da quantidade de fertilizantes aplicados no cultivo da cevada.

Efecto de un bioinoculante a partir de consorcios microbianos nativos fosfato solubilizadores, en el desarrollo de pastos Angleton (Dichantium aristatum) / Effect of bio-inoculant from microbial consortia phosphate solubilizing natives in development of pastures (Dichantium aristatum)

El objetivo fue evaluar la capacidad solubilizadora de fosfatos de consorcios formados por bacterias nativas de los géneros Burkholderia cepacia, Pseudomonas sp, Pseudomonas luteola y Pantoea sp, con el fin de encontrar el más eficiente. Se realizaron pruebas de antagonismo entre las cepas y se formaron consorcios probando todas las combinaciones posibles en las concentraciones de 10(6), 10(7), 10(8) UFC/mL. Se realizaron evaluaciones cualitativas y cuantitativas de la solubilización de fosfatos y teniendo en cuenta éstos resultados, se preparó un bioinoculante el cual fue evaluado en semillas de plantas de pasto angleton (Dichantium aristatum) a escala de laboratorio, utilizando un diseño estadístico completamente al azar (DCA) con 3 tratamientos y 5 repeticiones: Tratamiento 1 semillas (control), Tratamiento 2, semillas tratadas con el consorcio de microorganismos seleccionado y Tratamiento 3, semillas tratadas con fertilizantes comerciales DAP y Urea. Se evaluaron las variables número de hojas, área foliar, longitud de la planta, longitud de la raíz y peso seco de todas las plantas. Los resultados de la prueba de antagonismo indicaron que no existe inhibición en el crecimiento de las cepas evaluadas, por lo tanto se formaron consorcios los cuales mostraron mayor eficiencia en la solubilización del fósforo, destacándose el consorcio formado por Pantoea sp + Pseudomonas sp a una concentración de 10(8) UFC/mL y con índices de solubilización de 5,3 y 842 ppm. En las plantas se evidenció un incremento significativo en los parámetros peso seco y área foliar usando el consorcio microbiano, indicando mayor beneficio en comparación con el control.

The objective was to evaluate the ability of phosphate solubilizing consortium of native bacteria of the genusBurkholderia cepacia, Pseudomonas sp, Pseudomonas luteola and Pantoea sp, in order to find the most efficient. Antagonism tests were conducted between strains and consortia were formed using all possible combinations in the concentrations of 10(6), 10(7), 10(8) CFU / mL. Qualitative and quantitative determinations of the solubilization of phosphates were performed and considering these results, was prepared a bio-inoculant which was evaluated in plant seeds of grass angleton (Dichantium aristatum) laboratory scale, using a statistical completely randomized design (CRD) with 3 treatments and 5 repetitions: control Treatment 1 seeds; Treatment 2, seeds treated with the consortium of microorganisms selected and Treatment 3, seeds treated with commercial fertilizers DAP and Urea. The parameters, number of leaves, leaf area, plant length, root length and dry weights of all plants, were evaluated. The test results indicated that there is no antagonism inhibition in the growth of the strains tested thus formed consortia which showed greater efficiency phosphorus solubilization, highlighting the consortium of Pantoea sp + Pseudomonas sp at a concentration 10(8)CFU / mL and 5.3 solubilization rates and 842 ppm. In plants showed a significant increase in dry weight and leaf area parameters, indicating greater benefit with respect to the control treatment.

Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae) in the Brazilian Amazon / Bacterias diazotróficas aisladas de arroz silvestre Oryza glumaepatula (Poaceae) en la Amazonia brasileña

The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg’s medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH+ bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty- one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus. The growth- promoting ability was confirmed for at least five isolates. For these bacteria, the root and shoot growing results showed higher increases when compared to those observed in plants inoculated with the evaluated reference strains. These results indicate that O. glumaepatula is colonized by a high diverse diazotrophic community in the Brazilian Amazon. Further investigations are now being carried out to determine the taxonomic positions of these isolates and their growth promoting mechanisms.

La asociación de gramíneas silvestres con bacterias diazotróficas en los biomas brasileños es poco conocida. El aislamiento y caracterización de las bacterias asociadas con gramíneas silvestres puede contribuir a entender la ecología de las diazotróficas y bacterias con aplicaciones biotecnológicas. En este estudio, caracterizamos aislamientos bacterianos de diazotróficas de Oryza glumaepatula recolectadas en Cerrado y zonas boscosas de la Amazonía en el estado de Roraima, Brasil. Plantas sanas de O. glumepatula fueron recolectadas en cinco zonas boscosas y siete en Cerrado. Las plantas de Cerrado fueron recolectadas en septiembre 2008, mientras que las del bosque en Junio 2008 y Abril 2009. Las plantas y el suelo adherido a las raíces se transfirieron a macetas y se cultivaron durante 35 días en condiciones de invernadero. Durante la cosecha, los brotes y las raíces se trituraron por separado en una solución salina, la suspensión se diluyó en serie y se inocularon en placas Petri que contenían medio Dyg. Todas las colonias de bacterias se purificaron en el mismo medio. Se evaluó la capacidad diazotrófica de cada bacteria en condiciones microaerofílicas en medio semisólido BMGM. Además, los aislamientos bacterianos que formaron películas se evaluaron también mediante amplificación por PCR para el gen nifH. La diversidad de bacterias nifH+ se analizó por Huella Genética utilizando la Reacción en Cadena de la Polimerasa. Para las cepas seleccionadas, la capacidad de promover el crecimiento de O. sativa como modelo de planta también se evaluó. Se obtuvo un total de 992 cepas bacterianas. Cincuenta y un bacterias fueron capaces de formar películas en el medio semisólido y 38 amplificaron positivamente el fragmento 360bp del gen nifH. De los 38 aislamientos de nifH+, 24 fueron obtenidos de los brotes, mientras que 14 se originaron a partir de las raíces. Los perfiles de PCR-Box mostraron que los aislamientos bacterianos obtenidos en este estudio presentaron una baja similitud con las cepas de referencia pertenecientes a Herbaspirillum, Azospirillum y el género Burkholderia. La capacidad promotora del crecimiento fue confirmada por al menos cinco aislamientos. Para esta bacteria, la raíz y brote mostraron resultados de crecimiento mayores en comparación con los observados en las plantas inoculadas con las cepas de referencia. Estos resultados indican que O. glumaepatula es colonizada por una muy diversa comunidad diazotrófica en la Amazonia brasileña. Se están llevando a cabo otras investigaciones para esclarecer la taxonomía de estas cepas y sus mecanismos para promover el crecimiento.

Consumo e digestibilidade aparente de silagens de Brachiaria brizantha cv marandu adicionada de aditivos / Intake and digestibility of silages of Brachiaria brizantha cv marandu with additives

Avaliaram-se o consumo e a digestibilidade aparente da matéria seca, da proteína bruta e da energia e o balanço de nitrogênio de silagens de Brachiaria brizantha cvmarandusem aditivos (T1), B. brizantha cvmarandu + inoculante bacteriano Sil-ALL C4 (T2), B. brizantha cvmarandu + inoculante bacteriano Bactosilo C Tropical (T3) e B. brizantha cvmarandu + 30 por cento de cana-de-açúcar (T4) em ovinos. Não foram observadas diferenças no consumo de matéria seca (MS), proteína bruta (PB), energia bruta (EB), energia digestível (ED) e energia metabolizável (EM) entre os tratamentos (P>0,05). Os maiores valores de digestibilidade aparente da MS, PB e EB foram para o T2, de 60,9 por cento, 44,3 por cento e 57,5 por cento, respectivamente, e os menores para o T4, 53,0 por cento, 30,0 por cento e 49,5 por cento, respectivamente (P<0,05). Todos os tratamentos apresentaram balanço de nitrogênio positivo e não diferiram entre si. A utilização de inoculantes bacterianos ou cana-de-açúcar não indicou aumento de consumo da matéria seca e energia das silagens. O inoculante SiL ALL C4 (T2) mostrou-se eficiente em aumentar o consumo e a digestibilidade da PB.

This experiment was carried out to estimate the voluntary intake, apparent digestibility and nitrogen balance of silages of Brachiaria brizantha cv marandu without additives (T1), B. brizantha cv marandu + inoculant Sil-ALL C4 (T2), B. brizantha cv marandu + inoculant Bactosilo C Tropical (T3) and B. brizantha cv marandu + 30 percent of Sugar cane (T4) in sheep. No differences among silages regarding voluntary intake of: dry matter, crude protein, crude energy, digestible energy and metabolizable energy were found (P>0.05). The highest apparent digestibility of dry matter, crude protein and crude energy were observed for T2, being 60.8 percent, 44.2 percent and 57.5 percent, respectively, and the lowest for T4, with 53.0 percent, 30.0 percent and 49.5 percent, respectively (P<0.05). All treatments showed positive nitrogen balance and did not differ among them. The use of an inoculant or sugar cane did not result in voluntary intake increase of dry matter, fibrous fractions and energy of silages. The inoculant Sil ALL C4 was efficient in increasing the intake and digestibility of CP and nitrogen balance.

Utilization of rocks and ectomycorrhizal fungi to promote growth of eucalypt

The utilization of rocks as fertilizers is limited by their low solubility. However, solubilization may be achieved by some micro-organisms, such as ectomycorrhizal fungi (ECMf). The aim of this study was to evaluate the potential of seven isolates of ECMf to solubilize two rocks, alkaline breccia and granite, and to liberate potassium and phosphorus for Eucalyptus dunnii seedlings under greenhouse conditions. Fungal inoculants were produced in a peat-vermiculite-liquid medium mixture and added to the planting substrate at 10 percent. Rocks were ground up and added at 0.500 mg and 16.0 mg per plant, as a source of phosphorus and potassium, respectively. Other nutrients were added and E. dunnii seeds were sown. Control plants, non-inoculated, were fertilized with the same amount of phosphorus and potassium using soluble forms. After 90 days, the plant height, shoot dry weight, root length, phosphorus and potassium contents, and mycorrhizal colonization were evaluated. Alkaline breccia was more efficient than granite as a source of phosphorus and potassium for the plants, and may be an alternative to conventional fertilizers. Isolates UFSC-Pt22 (Pisolithus sp.) and UFSC-Pt186 (Pisolithus microcarpus) were the most efficient in promoting plant growth, mainly when combined with alkaline breccia to replace potassium and phosphorus fertilizers, respectively.

Germinação e vigor de sementes de arroz inoculadas com bactérias diazotróficas / Rice seed germination and vigour as affected by the inoculation with diazotrophic bacteria

Objetivou-se, neste trabalho, avaliar a germinação e o vigor das sementes de arroz inoculadas com bactérias diazotróficas. Foram instalados dois experimentos, sendo que no primeiro o delineamento experimental adotado foi o inteiramente casualizado em esquema fatorial, com dois lotes de sementes arroz da cultivar IC4440 x 10 tratamentos, com quatro repetições. Os tratamentos foram representados pela inoculação com oito estirpes de bactérias diazotróficas (AR1122, M130, BF1358, ZAE94, AR3122, AR2112, CD e BR2113), bem como pelo recobrimento das sementes com a turfa umedecida com o meio de cultivo esterilizado e pelas sementes não inoculadas. No segundo experimento, o delineamento experimental adotado foi o inteiramente casualizado, em esquema fatorial, com dois lotes de sementes recém-colhidas das cultivares IR42 e Zebu Branco x oito tratamentos, com quatro repetições. Os tratamentos foram representados pela inoculação das sementes com seis estirpes de bactérias diazotróficas (BF1358, AR3122, AR1122, ZAE94, M130 e AR2112), bem como pelo recobrimento das sementes com a turfa umedecida com o meio de cultivo esterilizado e pela não realização do procedimento de inoculação. As bactérias AR1122, M130, BF1358, ZAE94 e AR3112 podem ser selecionadas para formular inoculante para sementes de arroz. A inoculação com as bactérias diazotróficas aumenta a velocidade de germinação das sementes das cultivares IR42 e Zebu Branco, com menor contaminação por fungos.

The objective of this work was to evaluate the germination and vigour of rice seeds affected by inoculation with diazotrophic bacteria. Two experiments were conduced. In the first one, a completely randomized design with four replications arranged in a factorial scheme (2 lots x 10 treatments) was used. For this, rice seeds of cultivar IC4440 were inoculated with eight strains of diazotrophic (AR1122, M130, BF1358, ZAE94, AR3122, AR2112, CD e BR2113), submitted to coating with peat soil humidified with a sterile medium and not inoculated. In the second study, a completely randomized design with four replications arranged in a factorial scheme (2 lots x 6 treatments) was used. For this, recently harvested seeds of cultivars IR42 and Zebu Branco were inoculated with six strains (BF1358, AR3122, AR1122, ZAE94, M130 e AR2112), submitted to coating with peat soil humidified with a sterile medium and not inoculated. The bacteria AR1122, M130, BF1358, ZAE94 and AR3112 can be selected for inoculation of rice seeds. The inoculation of diazotrophic bacteria increases the speed of germination of rice seeds of cultivars IR42 e Zebu Branco, with low contamination by fungi.

Composição química e digestibilidade em ovinos da grama estrela ensilada com diferentes aditivos / Chemical composition and digestibility in sheep of stargrass ensiled with different additives

Objetivou-se com o estudo avaliar a composição química e a digestibilidade aparente em ovinos da grama estrela ensilada com diferentes aditivos. Os aditivos empregados foram o enzimo-bacteriano (em duas concentrações) e a uréia (10 kg/tonelada), totalizando quatro tratamentos. Para o experimento de digestibilidade, foram utilizados quatro ovinos machos castrados distribuídos em delineamento quadrado latino (4x4). Em relação à composição química, as variáveis MS, PB, EE, FDA e LIG apresentaram diferença (P<0,05) entre os valores obtidos, porém a FDN não mostrou diferença entre tratamentos. Quanto à capacidade tampão e ao pH os menores valores foram obtidos com os tratamentos com aplicação de inoculante. Não houve efeito de tratamento (inoculante ou aplicação de uréia) sobre a digestibilidade da MS, PB e FDN.

The purpose of this study was to evaluate the chemical composition and apparent digestiblity in sheep of stargrass ensilaged with different additives. The additives used were enzyme-bacterial inoculent (in two concentrations) and urea (10 kg/ton), totalling 4 treatments. For the digestability experiment, four male sheep were distributed in a (4x4) latin square design. With respect to chemical composition, the variables; dry matter, crude protein, ether extract, acid detergent fiber, and lignin showed statistical difference (P<0,05) among the obtained values. However, neutral detergent fiber showed no difference among treatments. Concerning buffering capacity and pH, the lowest values were for silages treated with inoculents. No treatment effects (inoculent or urea application) for dry matter, crude protein, or neutral detergent fiber digestibility of stargrass silages were detected.

Consumo e digestibilidade de nutrientes de dietas com silagens de grãos úmidos de milho ou sorgo, em ovinos / Intake and digestibility of nutrients in sheep diets based on corn and sorghum grain high moisture silages

Avaliaram-se o consumo, a digestibilidade e o teor de nutrientes digestíveis totais (NDT) de dietas contendo silagens de grãos úmidos de milho e de sorgo, com ou sem inoculante microbiano, em ovinos. O volumoso utilizado foi feno de capim-braquiária (Brachiaria brizantha cv. Marandu). Doze animais foram distribuídos em quatro tratamentos: (1) feno+silagem de grão úmido de milho, (2) feno+silagem de grão úmido de sorgo; (3) feno+silagem de grão úmido de milho com inoculante microbiano e (4) feno+silagem de grão úmido de sorgo com inoculante microbiano. Não houve diferença entre tratamentos quanto aos consumos de matéria seca, de matéria orgânica, de proteína bruta, de fibra em detergente neutro, de carboidratos totais e de carboidratos não estruturais. O consumo de extrato etéreo diferiu em função do tipo de grão. Não houve diferença entre tratamentos quanto à digestibilidade dos nutrientes. Não houve efeito do grão e da inoculação para o NDT estimado das silagens de grãos úmidos de milho e de sorgo, com médias de 77,6 por cento e 79,5 por cento, respectivamente. O uso de inoculante microbiano em silagens de grãos úmidos não se fez necessário. O grão de milho úmido pode ser substituído pelo de sorgo sem prejuízos ao aproveitamento de nutrientes em ovinos adultos.

The intake, the digestibility of nutrients, and the total digestible nutrients (TDN) content in sheep diets based on corn and sorghum grains high moisture silages with or without microbial inoculant, were evaluated. The used roughage was Brachiaria hay (Brachiaria brizantha cv. Marandu). Twelve animals were distribuited to treatments: (1) hay+high moisture corn grain silage, (2) hay+high moisture sorghum grain silage; (3) hay+high moisture corn grain silage with microbial inoculant, and (4) hay+high moisture sorghum grain silage with microbial inoculant. No differences among treatments in relation to the intake of dry matter, organic matter, crude protein, neutral detergent fiber, and total and non-structural carbohydrates were observed. The ether extract intake differed according to the grains. The silages did not differ in relation to the estimated digestibilities for dry matter, organic matter, crude protein, neutral detergent fiber, ether extract, and total and non-structural carbohydrates. The TDN did not differ according to the grains and inoculant, with averages of 77.6 and 79.5 percent, for corn and sorghum grains high moisture silages, respectively. It was demonstrated that the use of microbial inoculant, in high moisture grain silages was not necessary. High moisture corn grain silage can be substituted by high moisture sorghum grain silage without effects on the use of nutrients by sheep.

Degradabilidade ruminal e fracionamento de carboidratos de silagens de grama estrela (Cynodon nlemfuensis vanderyst. ) com diferentes aditivos / Ruminal degradability and carbohydrate fractioning of stargrass silages (Cynodon nlemfuensis vanderyst. ) with different additives

Objetivou-se com o estudo avaliar o efeito de aditivo enzimo-bacteriano e de uréia sobre a degradabilidade ruminal e o fracionamento de carboidratos em silagens de grama estrela. Foram avaliadas silagens sem aditivo (SGE) e com aplicação de aditivo enzimo-bacteriano (SGE150 e SE300, respectivamente 150 e 300g aditivo/tonelada) e uréia (SGEU, 10 kg/tonelada). Para o ensaio de degradabilidade foram utilizados três bois da raça Holandesa distribuídos em blocos casualizados. A degradabilidade efetiva (DE) da MSe da FDN foram maiores para o tratamento SGE300 em todas as taxas de passagem. Para a DE da PB houve diferença (P<0,05) para o tratamento SGEU. Os carboidratos totais (CHT) tiveram os maiores valores para o tratamento SGE, enquanto que, para as frações A+B1 e B2 não houve efeito significativo entre os tratamentos. Para a fração C os menores valores foram para os tratamentos com inoculante. Conclui-se que a aplicação do inoculante ou uréia não melhoraram a degradabilidade potencial (DP) das silagens de grama estrela em relação ao tratamento isento da aplicação de inoculante.

It was aimed to evaluate in this trial the effect of additive and urea supplies on ruminal degradabilty andcarbohydrates fractioning of stargrass silages. There were evaluated silage without additive (SGE),silage with enzymatic-bacterial additive (SGE150 and SGE300 respectively, 150 and 300 g of additive/tonof silage) and silage with urea (SGEU, 10 kg/ton). Three Holstein steers were used in blocks randomizedsdesign for ruminal degradability assay. The effective degradability (ED) of DM and NDF were higher(P<0.05) for SGE300 treatment for all flow rates. There was difference (P<0.05) for ED of CP in silagesupplied with urea. Total carbohydrates presented the highest values for no additive treatment, whileA+B1 and B2 fractions didn’t differ (P>0.05) among treatments. Silages with additive presented thelowest values (P<0.05) of C fraction. It was concluded that additive or urea supply in stargrass silagesdidn’t improve the potential degradability (PD) when compared to the silage without additive.

Bacteria Composition of Organic Matter-decomposing Inoculant Analyzed with 16S rDNA Clone Library / 微生物学通报

Using 16S rDNA clone library method, the bacteria composition of the Organic Matter-decomposing Inoculants A and B were investigated. The results indicated that: Sample A was clustered into 14 taxonomic operational units (OTUs),the dominant communities are Weissella confuse, Bacillus subtilis and Bacillus pumilus, which accounted for 28.6%,30.4% and 23.2%;Sample B was clustered into 43 OTUs,the dominant communities are Lactobacillus buchneri, Lactobacillus farciminis and Lactobacillus acetotolerans, which accounted for 18.03%,18.86% and 13.12%, respectively. The results had much difference from the samples' labels:there were not bacteria in the lable of sample A and only Bacillus pumilus was the same with the lable of sample B.This study showed that 16S rDNA clone library method has nicer application perspectives in analying the bacteria of microbial inoculant and its quality control.