Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2016-08-29T12:34:04Z No. of bitstreams: 1 PDF - Dalila Ribeiro Rodrigues.pdf: 1923244 bytes, checksum: f45088c1a8a811273f1ac7c37c1e3aec (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-08-29T17:02:09Z (GMT) No. of bitstreams: 1 PDF - Dalila Ribeiro Rodrigues.pdf: 1923244 bytes, checksum: f45088c1a8a811273f1ac7c37c1e3aec (MD5) / Made available in DSpace on 2016-08-29T17:02:09Z (GMT). No. of bitstreams: 1 PDF - Dalila Ribeiro Rodrigues.pdf: 1923244 bytes, checksum: f45088c1a8a811273f1ac7c37c1e3aec (MD5) Previous issue date: 2016-04-01 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The use of legumes inoculated with competitive and efficient nitrogen-fixing bacteria is a strategy to enhance the practices in the regeneration of degraded areas. In addition to encouraging the establishment of plants, the use of efficient strains reduces the cost of deployment and maintenance of agricultural systems, eliminating the use of nitrogenous fertilizers. But for the success of these practices is necessary to know the diversity of bacteria in the legumes nodules and evaluate their proper efficiency. The objective of this study was to evaluate the diversity and efficiency of bacterial isolates from three native species nodules Caatinga, grown in the State of Pernambuco soils deposited in the Agricultural Interest Collection of Microorganisms of Embrapa Semi -Arid. After certification purity of the stock of bacteria, all 308 isolates passed through a selection of amplicons nodC and nifH genes using a duplex-PCR approach. Among the bacteria evaluated, positive amplification were obtained for 18 isolated from mulungu, 40 to jurema-preta and 44 to angico. All these isolates were evaluated for NaCl tolerance and high temperatures "in vitro", ability to metabolize different carbon sources and enzymatic activity (only bacteria from mulungu), their “in vitro” plant growth promotion mechanisms by means of the production of indole acetic aci d (IAA) and calcium phosphate solubilization and finally tested in the greenhouse with the original 2 host plants, to determine the symbiotic efficiency of the isolates. Isolates of mulungu showed variability for enzyme activity and metabolism of carbon sources. Five isolated mulungu and jurema-preta and 6 isolates from angico were able to produce IAA in L-tryptophan supplemented medium, reaching rates above those achieved by the reference strain. As the potential of calcium phosphate solubilization for mulungu isolates, only one bacteria was able to achieve high solubilization equal to the reference strain, seven and two isolates from jurema-preta and angico, solubilized calcium phosphate above the reference strains value. For each species there bacteria were able to show tolerance to high temperatures and salinity. Isolates of angico and jurema were submitted to ARDRA using three endonucleases: MspI, HinfI and HhaI. The genetic diversity evaluation indicated that to angico a large cluster with around 40% of similarity was formed. To jurema-preta a major cluster with 65% of similarity among the isolates were achieved. In addition to high diversity of bacteria, patterns regarding its geographical occurrence were also observed. Bacteria from mulungu had the sequences of the 16S rRNA gene partially determinated. Among the 13 bacteria with good sequences, six belong to Rhizobium, six Bradyrhizobium and one belongs to Burkholderia. The Efficiency of tests, 22 isolates of mimosa and 10 mulungu were able to renodulate their original host strains and A27 from Angico and M14 and M31 can be highlighted to mulungu. / A utilização de leguminosas inoculadas com bactérias fixadoras de nitrogênio nativas eficientes e competitivas representa uma estratégia para potencializar as práticas na regeneração de áreas degradadas. Além de favorecer o estabelecimento das plantas, a utilização de estirpes eficientes reduz os custos da implantação e manutenção dos sistemas agrícolas, dispensando a utilização de fertilizantes químicos nitrogenados. Mas, para o sucesso destas práticas é preciso conhecer a diversidade das bactérias presentes nos nódulos de tais espécies de leguminosas e sua devida eficiência. O objetivo deste trabalho foi avaliar a diversidade e eficiência de bactérias oriundas de solos do Estado de Pernambuco e isoladas de nódulos de três espécies leguminosas arbóreas nativas da Caatinga, depositadas na Coleção de Microorganismos de Interesse Agrícola da Embrapa Semiárido. Após certificação da pureza do estoque de bactérias, os isolados passaram por uma seleção através da amplificação de fragmentos dos genes nodC e nifH utilizando a abordagem de duplex PCR. Apresentaram amplificação positiva 18 isolados de mulungu, 40 de jurema-preta e 44 de angico, esses isolados foram avaliados quanto a tolerância de NaCl e altas temperaturas “in vitro”, habilidade de metabolizar diferentes fontes de carbono e atividade enzimática (apenas isolados de mulungu), sua capacidade de promoção do crescimento vegetal, através da produção do Ácido indol-acético (AIA) e solubilização de fosfato de cálcio e, por fim, foram testados em casa de vegetação com as plantas hospedeiras originais, para determinação da eficiência simbiótica. Os isolados do mulungu mostraram variabilidade para atividade enzimática e para metabolização das fontes de carbono. Cinco isolados de mulungu e jurema-preta e 6 isolados de angico foram capazes de produzir AIA, na presença do L-triptofano, acima da estirpe de referência. Quanto ao potencial de solubilização de fosfato de cálcio apenas um isolado de mulungu apresentou valor igual a estirpe de referência, sete isolados de jurema-preta e dois de angico, solubilizaram fosfato de cálcio acima do valor da referência. Para cada espécie existem representantes potenciais na tolerância de altas temperaturas e altos teores salinos. Os isolados de angico e jurema-preta com características promíscuas foram submetidos ao ARDRA, utilizando três enzimas de restrição MspI, HinfI e HhaI, gerando para o angico um grande grupo com aproximadamente 40% de similaridade e para juremapreta um principal grupo com 65% de similaridade entre os isolados, constatando então a presença da diversidade dentre os isolados. Nos testes de autenticidade, 22 isolados de angico e 10 de mulungu foram capazes de renodular a estirpe hospedeira e o isolado A27, é eficiente na fixação biológica de nitrogênio, quando comparado ao controle nitrogenado. As bactérias do mulungu que se destacaram tiveram o gene 16S do rRNA parcialmente sequenciado para determinar o posicionamento taxonômico.

A adoção da rotação de culturas com leguminosas em áreas de implantação e reforma de canaviais é uma estratégia promissora para recuperação e manutenção da fertilidade do solo, e tem potencial no aproveitamento do N para as soqueiras. A análise mais detalhada do comportamento do N no solo durante o ano agrícola e dos parâmetros de produção em locais que possuem solo e clima contrastantes, permitem aprimorar o manejo da adubação nitrogenada da cana-de-açúcar. As hipóteses deste trabalho são (1) a rotação de culturas irá aumentar a disponibilidade de N no solo, a qual suprirá parte da demanda do N das soqueiras subsequentes e (2) a rotação de culturas promoverá redução da resposta das soqueiras à adubação nitrogenada. Objetivou-se avaliar o efeito da rotação de culturas nas transformações do N no solo (N mineral, N-ISNT e N e C da biomassa microbiana) e na produtividade da primeira soqueira. Os experimentos foram instalados em quatro regiões representativas de cultivo de cana-de-açúcar na região Centro Sul do Brasil: Quatá/SP (Local A); Iracemápolis/SP (Local B); Chapadão do Céu/GO (Local C) e Quirinópolis/GO (Local D). Em cada local, foi instalado um experimento em esquema de parcelas subdivididas no delineamento blocos ao acaso com quatro repetições, em áreas submetidas (com rotação) ou não (sem rotação) ao plantio de leguminosa do gênero crotalária. Após a colheita da cana planta, foram implantados os seguintes tratamentos: 60, 120 e 180 kg ha-1 de N e um controle (sem N), aplicados superficialmente sobre a palhada. Foram realizadas três amostragens para determinação do conteúdo de N mineral, N-ISNT e C e N da biomassa microbiana. Entre 150 a 180 dias após a colheita da cana-planta, foram realizadas avaliações de perfilhamento e índice SPAD. Na colheita, avaliou-se matéria seca da parte aérea, produtividade e atributos tecnológicos. Os resultados demonstraram que a mineralização dos resíduos da crotalária influenciaram a dinâmica do C e N e os reservatórios de N no perfil do solo ao longo do ciclo da primeira soqueira, com destaque para o solo de textura média (Local A). Apesar da rotação de culturas ter modificado o conteúdo de N mineral, N-ISNT, e aumentado o perfilhamento e índice SPAD no local A, somente nos solos argilosos (Locais C e D), a rotação aumentou a produtividade da soqueira. A hipótese de que a rotação de culturas reduz a demanda de fertilizantes nitrogenados não foi aceita; houve resposta à adubação nitrogenada nos quatro locais, independentemente da rotação de culturas. Porém, a rotação demonstrou potencial em aumentar a produção de biomassa com manutenção da dose de N. Esta pesquisa demonstrou a viabilidade de rotação de culturas em solos argilosos com elevado conteúdo de C e N total, com potencial de aumentar a produtividade do canavial, porém sem reduzir a demanda de adubação nitrogenada. / The adoption of crop rotation with sugarcane is a promising strategy for the recovery and maintenance of soil fertility, and it shows potential in the use of N to the sugarcane ratoon. A more detailed analysis of the transformations N in soil during the agricultural year and production parameters in sites that have contrasting soil texture and climate, allows to improve management of N fertilization for sugarcane. The hypothesis of this work are (1) crop rotation will increase soil N availability, which will satisfy part of N requirements of sugarcane ratoon and (2) crop rotation will reduce the sugarcane ratoon response to N fertilization. The objective was to evaluate the effects of sunn hemp rotation in the transformations of N in soil (mineral N content, ISNT-N and microbial biomass C and N) and sugarcane ratoon yield. The field trials were installed in four representative regions of cultivation of sugarcane in Brazil: Quatá/SP (Site A); Iracemápolis/SP (Site B); Chapadão do Céu/GO (Site C) e Quirinópolis/GO (Site D). At each site, a field trial was installed in split plot in a randomized block experimental design with four replications, in areas sowed (rotation) and not sowed (no rotation) with sunn hemp. After the harvest of cane-plant, the following treatments were applied: 60, 120 and 180 kg ha-1 of N and a control (without N) over the straw. Three sampling were performed to determine the soil inorganic N content, ISNT-N, C and N of microbial biomass. Between 150 to 180 days after the harvest of the cane-plant, tillering was counted and SPAD index was measured. At harvest of the sugarcane ratoon, biometric parameters, yield and quality parameters were evaluated. The results showed that mineralization of residues of sunn hemp influenced the dynamics of C and N and N content in the soil profile throughout the cycle of the sugarcane ratoon, with emphasis on medium-textured soil (Site A). Despite crop rotation has modified mineral N stocks, ISNT-N, and increased tillering and SPAD index in site A, crop rotation only increased the sugarcane ratoon yield on clay soils (Sites C and D). The hypothesis that crop rotation reduces N-fertilizer requirement was not accepted; there was response to N fertilization in four sites, regardless of crop rotation. However, the rotation showed potential in increase biomass production with maintenance of N rate. This research demonstrated the viability of crop rotation in clay soils with high total C and N content, with potential to increase sugarcane yield, but without reducing N-fertilizer requirement.

Made available in DSpace on 2017-07-10T17:37:23Z (GMT). No. of bitstreams: 1 Jesica_F_S_Genero.pdf: 1260489 bytes, checksum: da1efddef812f7eb843220dbd5d18a24 (MD5) Previous issue date: 2016-08-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The bacteria Azospirillum brasilense and Herbaspirillum seropedicae are nitrogen fixers and promoting plant growth, able to promote partnerships with the trigo. O objective of this study was to evaluate the crop wheat agronomic performance in response to inoculation with Azospirillum brasilense diazotrophs and Herbaspirillum seropedicae with or without nitrogen fertilization and quantify the number of viable cells of bacteria of the wheat plant roots. The experiment was conducted in the crop year 2014/2014 and 2015/2015. The experimental design was a randomized block in factorial 2 x 4, with four repetitions. The first factor relates to the nitrogen topdressing applications (with and without nitrogen). The second factor relates to inoculating seeds with diazotrophs [without inoculation treatment (control), strain A. brasilense, H. seropedicae strain and combination of the two strains. The combined inoculation of strains A. brasilense + H. seropedicae with nitrogen fertilization promoted an increase of 48% and 13% in grain yield, in the 2014 season and 2015 season, respectively, for the plants that were not inoculated. You can check the inoculation Herbaspirillum, showed better total amount of viable cells with the application of nitrogen. The reduction in the number of CFU of Azospirillum under nitrogen fertilization condition can point to a lower competitiveness of these individuals by exudates and nutrients / As bactérias Azospirillum brasilense e Herbaspirillum seropedicae são fixadoras de nitrogênio e promotoras de crescimento vegetal, capazes de promover associações com a cultura do trigo. O objetivo deste trabalho foi avaliar o desempenho agronômico do trigo em resposta a inoculação de bactérias diazotróficas com Azospirillum brasilense e Herbaspirillum seropedicae associada ou não à adubação nitrogenada e quantificar o número de células viáveis de bactérias das raízes de plantas de trigo. O experimento foi realizado nas safras agrícolas 2014/2014 e 2015/2015. O delineamento experimental foi de blocos casualizados em esquema fatorial 2 x 4, com 4 repetições. O primeiro fator refere-se às aplicações de nitrogênio em cobertura (com e sem nitrogênio). O segundo fator refere-se à inoculação de sementes com bactérias diazotróficas [tratamento sem inoculação (testemunha), estirpe de A. brasilense, estirpe de H. seropedicae e combinação das duas estirpes. Na ausência da inoculação, a adubação nitrogenada em cobertura promoveu um acréscimo de 18% no diâmetro do colmo. Na ausência da adubação nitrogenada em cobertura, a inoculação com H. seropedicae promoveu um acréscimo de 16% no diâmetro de colmo, e na presença da adubação nitrogenada esta bactéria promoveu um acréscimo de 24% no número de espiguetas por espiga em relação às plantas que não foram inoculadas. A inoculação combinada das estirpes A. brasilense + H. seropedicae com adubação nitrogenada em cobertura promoveu um acréscimo de 26% no número de espiguetas por espiga e o dobro do número de grãos obtidos, em relação à combinação sem adubação. A inoculação combinada das estirpes A. brasilense + H. seropedicae com adubação nitrogenada em cobertura promoveu um acréscimo de 48% e 13% na produtividade de grãos, na safra de 2014 e na safra de 2015, respectivamente, em relação as plantas que não foram inoculadas. A inoculação de H. seropedicae apresentou maior quantidade de células viáveis totais com a aplicação de nitrogênio. A redução do número de UFC de A. brasilense sob condição de adubação nitrogenada pode apontar para uma menor competitividade destes indivíduos por exsudatos e nutrientes

A adoção da rotação de culturas com leguminosas em áreas de implantação e reforma de canaviais é uma estratégia promissora para recuperação e manutenção da fertilidade do solo, e tem potencial no aproveitamento do N para as soqueiras. A análise mais detalhada do comportamento do N no solo durante o ano agrícola e dos parâmetros de produção em locais que possuem solo e clima contrastantes, permitem aprimorar o manejo da adubação nitrogenada da cana-de-açúcar. As hipóteses deste trabalho são (1) a rotação de culturas irá aumentar a disponibilidade de N no solo, a qual suprirá parte da demanda do N das soqueiras subsequentes e (2) a rotação de culturas promoverá redução da resposta das soqueiras à adubação nitrogenada. Objetivou-se avaliar o efeito da rotação de culturas nas transformações do N no solo (N mineral, N-ISNT e N e C da biomassa microbiana) e na produtividade da primeira soqueira. Os experimentos foram instalados em quatro regiões representativas de cultivo de cana-de-açúcar na região Centro Sul do Brasil: Quatá/SP (Local A); Iracemápolis/SP (Local B); Chapadão do Céu/GO (Local C) e Quirinópolis/GO (Local D). Em cada local, foi instalado um experimento em esquema de parcelas subdivididas no delineamento blocos ao acaso com quatro repetições, em áreas submetidas (com rotação) ou não (sem rotação) ao plantio de leguminosa do gênero crotalária. Após a colheita da cana planta, foram implantados os seguintes tratamentos: 60, 120 e 180 kg ha-1 de N e um controle (sem N), aplicados superficialmente sobre a palhada. Foram realizadas três amostragens para determinação do conteúdo de N mineral, N-ISNT e C e N da biomassa microbiana. Entre 150 a 180 dias após a colheita da cana-planta, foram realizadas avaliações de perfilhamento e índice SPAD. Na colheita, avaliou-se matéria seca da parte aérea, produtividade e atributos tecnológicos. Os resultados demonstraram que a mineralização dos resíduos da crotalária influenciaram a dinâmica do C e N e os reservatórios de N no perfil do solo ao longo do ciclo da primeira soqueira, com destaque para o solo de textura média (Local A). Apesar da rotação de culturas ter modificado o conteúdo de N mineral, N-ISNT, e aumentado o perfilhamento e índice SPAD no local A, somente nos solos argilosos (Locais C e D), a rotação aumentou a produtividade da soqueira. A hipótese de que a rotação de culturas reduz a demanda de fertilizantes nitrogenados não foi aceita; houve resposta à adubação nitrogenada nos quatro locais, independentemente da rotação de culturas. Porém, a rotação demonstrou potencial em aumentar a produção de biomassa com manutenção da dose de N. Esta pesquisa demonstrou a viabilidade de rotação de culturas em solos argilosos com elevado conteúdo de C e N total, com potencial de aumentar a produtividade do canavial, porém sem reduzir a demanda de adubação nitrogenada. / The adoption of crop rotation with sugarcane is a promising strategy for the recovery and maintenance of soil fertility, and it shows potential in the use of N to the sugarcane ratoon. A more detailed analysis of the transformations N in soil during the agricultural year and production parameters in sites that have contrasting soil texture and climate, allows to improve management of N fertilization for sugarcane. The hypothesis of this work are (1) crop rotation will increase soil N availability, which will satisfy part of N requirements of sugarcane ratoon and (2) crop rotation will reduce the sugarcane ratoon response to N fertilization. The objective was to evaluate the effects of sunn hemp rotation in the transformations of N in soil (mineral N content, ISNT-N and microbial biomass C and N) and sugarcane ratoon yield. The field trials were installed in four representative regions of cultivation of sugarcane in Brazil: Quatá/SP (Site A); Iracemápolis/SP (Site B); Chapadão do Céu/GO (Site C) e Quirinópolis/GO (Site D). At each site, a field trial was installed in split plot in a randomized block experimental design with four replications, in areas sowed (rotation) and not sowed (no rotation) with sunn hemp. After the harvest of cane-plant, the following treatments were applied: 60, 120 and 180 kg ha-1 of N and a control (without N) over the straw. Three sampling were performed to determine the soil inorganic N content, ISNT-N, C and N of microbial biomass. Between 150 to 180 days after the harvest of the cane-plant, tillering was counted and SPAD index was measured. At harvest of the sugarcane ratoon, biometric parameters, yield and quality parameters were evaluated. The results showed that mineralization of residues of sunn hemp influenced the dynamics of C and N and N content in the soil profile throughout the cycle of the sugarcane ratoon, with emphasis on medium-textured soil (Site A). Despite crop rotation has modified mineral N stocks, ISNT-N, and increased tillering and SPAD index in site A, crop rotation only increased the sugarcane ratoon yield on clay soils (Sites C and D). The hypothesis that crop rotation reduces N-fertilizer requirement was not accepted; there was response to N fertilization in four sites, regardless of crop rotation. However, the rotation showed potential in increase biomass production with maintenance of N rate. This research demonstrated the viability of crop rotation in clay soils with high total C and N content, with potential to increase sugarcane yield, but without reducing N-fertilizer requirement.

Stable isotope probing (SIP) is an established technique that can be applied to identify the metabolically active micro-organisms within a microbial population. The SIP method utilises an isotopically-labelled substrate and PCR techniques to discern the members of a microbial community that incorporate the isotope into their DNA or RNA. The current literature gap around using 15N isotopes with RNA-SIP offers real potential and advantages for targeting and identifying active members from mixed communities involved in global biogeochemical nitrogen cycling. This study specifically investigated whether nitrogen based compounds can be used as substrates in RNA-SIP methodologies and whether they can in turn be used to probe mixed community environments known to be actively fixing nitrogen. The nitrogen-limited systems targeted represented an ideal opportunity to assess the suitability of 15N-RNA-SIP approaches due to their known high nitrogen fixation rates. Identifying these nitrogen-fixing bacteria could provide a better representation analysis of the community, leading to an improved prediction on how to manage and optimise the treatment performance of target waste systems and to exploit the unique bioconversion properties of these types of organisms. Initially, the project undertook methodological proof of concept by using a soluble nitrogen source, 15NH4Cl, to label the RNA of Novosphingobium nitrogenifigens and a mixed microbial community. Successful separation of the 14N- (control) and 15N-RNA was achieved for both pure and mixed communities using isopycnic caesium trifluoroacetate (CsTFA) gradients in an ultracentrifuge. The usefulness of this technique to identify active diazotrophs in real environmental samples was tested using a nitrogen-fixing community from a pulp and paper wastewater treatment system. After growing the mixed culture with 15N2 as the sole nitrogen source, the labelled RNA was extracted and fractionated using isopycnic centrifugation in CsTFA gradients. The community composition of the active nitrogen-fixing community in the 15N2 enriched fraction was analysed by establishing a 16S rRNA gene clone library containing over 200 members. These were analysed by comparison with published sequences and by phylogenetic analysis. It was found that the more isotopic label substrate incorporated, the further the buoyant density (BD) separation between 15N- and 14N-RNA. Novosphingobium nitrogenifigens gave an average BD shift of 0.03 + 0.004 g ml-1 (95.0 atom % 15N) with 15NH4Cl. For mixed communities the average BD shift was 0.02 + 0.004 g ml-1 (80.0 atom % 15N) with 15NH4Cl and 0.013 + 0.002 g ml-1 (32.6 atom % 15N) when using 15N2. Clone library analysis of 16S rRNA genes present in the enriched 15N-RNA fraction of the mixed community was shown to consist of a diverse population of bacteria as indicated by a Shannon Weaver index value of gt;2.8. Three dominant genera (Aeromonas, Pseudomonas and Bacillus) were identified by comparison with published sequences and phylogenetic analysis. Many other groups not known as archetypal nitrogen-fixing bacteria were also identified, demonstrating that 15N2-RNA-SIP provides a useful tool for the identification of important and previously unknown contributors to nitrogen fixation in a range of environments. Overall, this project has established that nitrogen based RNA-SIP is a powerful tool that can be used successfully and reproducibly with both pure and complex mixed microbial communities to study active diazotrophs in environmental samples.

On the Canadian prairies, organic production generally includes the use of annual green manure (GrM) crops, which are terminated using tillage to add nutrients and organic matter to the soil. However, in a GrM plough-down year, farmers face loss of income. As an alternative to growing traditional GrM crops, legumes can be grown alone or intercropped with cereals and harvested as green feed forage (GF) for use on-farm or for sale to other producers without depleting soil nitrogen (N) for the subsequent crop. We hypothesized that the GF system would have similar biomass, and N yield, and ultimately would return N into the soil. Furthermore, by intercropping a legume with a cereal, biological N2-fixation will be enhanced in the legume.<p> Field experiments, conducted over two years, were established at Vonda and Delisle, Saskatchewan, Canada. The experiment was conducted using a randomized complete block design (RCBD) with 16 treatments and four replicates in which field pea (<i>Pisum sativum</i> cv 40-10 silage pea), oat (<i>Avena sativa</i> L.cv AC Morgan), and triticale (X <i>Triticosecale</i> Wittmack cv Pika) were grown alone or in combination and managed as GrM or GF. Wheat and tillage fallow served as cropped and uncropped controls, respectively. The tillage fallow-control system was tilled twice in the growing season using a small tractor disc. The intercropped oat was seeded at three densities (50, 100, and 150 plants m-2) to determine whether increasing cereal density stimulated N2-fixation in the field pea.<p> The GrM system was sampled and incorporated (when the field pea was at full bloom) two weeks earlier than the GF system. Consequently, at both sites, all treatments in the GF system consistently yielded more dry matter and accumulated more N than treatments in the GrM system. At the Delisle site, where percent nitrogen derived from the atmosphere (%Ndfa) was compared, increasing cereal density did not increase N2-fixation in both management systems. However, pea in the GF system accumulated more than twice the amount of N (kg ha-1) from fixation as compared to pea in the GrM system, presumably because of the longer growth period.<p> Wheat grown following the GrM treatments produced more biomass and accumulated more N than wheat following the GF treatments. Wheat grown after the monoculture field pea as a GrM had greater yield than all treatments. As well, the GrM system returned more N to the soil than did the GF system. The extra two weeks of growth in the GF system resulted in the extraction of significant amounts of nutrients and probably moisture from the soil, which adversely affected yield and nutrient composition of the following wheat crop.<p> Although organic farmers may lose income in the plough-down year, on a longterm soil sustainability basis, the GrM system is a better option than the GF system as it returns nutrients to the soil, thus providing improved plant biomass, and N accumulation of subsequent crops. However, organic farmers growing GF for hay may benefit from the increased productivity of this system on a short-term basis. Thus, farmers pursuing GF options may need to adopt other means of sustaining soil productivity on a longer term. The tilled fallow-control system resulted in high amounts of biomass and N accumulation by the subsequent wheat crop, probably due to the fact that there were no nutrients taken up in the previous year and moisture was conserved in these treatments. However, this system may have less long-term benefits compared to the GrM regime, as no nutrients are returned through ploughing down a crop.

On the Canadian prairies, organic production generally includes the use of annual green manure (GrM) crops, which are terminated using tillage to add nutrients and organic matter to the soil. However, in a GrM plough-down year, farmers face loss of income. As an alternative to growing traditional GrM crops, legumes can be grown alone or intercropped with cereals and harvested as green feed forage (GF) for use on-farm or for sale to other producers without depleting soil nitrogen (N) for the subsequent crop. We hypothesized that the GF system would have similar biomass, and N yield, and ultimately would return N into the soil. Furthermore, by intercropping a legume with a cereal, biological N2-fixation will be enhanced in the legume.<p> Field experiments, conducted over two years, were established at Vonda and Delisle, Saskatchewan, Canada. The experiment was conducted using a randomized complete block design (RCBD) with 16 treatments and four replicates in which field pea (<i>Pisum sativum</i> cv 40-10 silage pea), oat (<i>Avena sativa</i> L.cv AC Morgan), and triticale (X <i>Triticosecale</i> Wittmack cv Pika) were grown alone or in combination and managed as GrM or GF. Wheat and tillage fallow served as cropped and uncropped controls, respectively. The tillage fallow-control system was tilled twice in the growing season using a small tractor disc. The intercropped oat was seeded at three densities (50, 100, and 150 plants m-2) to determine whether increasing cereal density stimulated N2-fixation in the field pea.<p> The GrM system was sampled and incorporated (when the field pea was at full bloom) two weeks earlier than the GF system. Consequently, at both sites, all treatments in the GF system consistently yielded more dry matter and accumulated more N than treatments in the GrM system. At the Delisle site, where percent nitrogen derived from the atmosphere (%Ndfa) was compared, increasing cereal density did not increase N2-fixation in both management systems. However, pea in the GF system accumulated more than twice the amount of N (kg ha-1) from fixation as compared to pea in the GrM system, presumably because of the longer growth period.<p> Wheat grown following the GrM treatments produced more biomass and accumulated more N than wheat following the GF treatments. Wheat grown after the monoculture field pea as a GrM had greater yield than all treatments. As well, the GrM system returned more N to the soil than did the GF system. The extra two weeks of growth in the GF system resulted in the extraction of significant amounts of nutrients and probably moisture from the soil, which adversely affected yield and nutrient composition of the following wheat crop.<p> Although organic farmers may lose income in the plough-down year, on a longterm soil sustainability basis, the GrM system is a better option than the GF system as it returns nutrients to the soil, thus providing improved plant biomass, and N accumulation of subsequent crops. However, organic farmers growing GF for hay may benefit from the increased productivity of this system on a short-term basis. Thus, farmers pursuing GF options may need to adopt other means of sustaining soil productivity on a longer term. The tilled fallow-control system resulted in high amounts of biomass and N accumulation by the subsequent wheat crop, probably due to the fact that there were no nutrients taken up in the previous year and moisture was conserved in these treatments. However, this system may have less long-term benefits compared to the GrM regime, as no nutrients are returned through ploughing down a crop.

Whole farm nutrient budgets (WFNB) enable producers to link dairy herd management with traditional field nutrient management plans. The objective of this study was to calculate WFNBs of nitrogen, phosphorus, and potassium (N, P, and K) at a commercial farm in New Brunswick and in Prince Edward Island. Reliable estimates of N fixation from alfalfa and red clover on the farms were obtained with adjustments to the Høgh-Jensen et al. (2004) dry matter conversion models. The farms had surpluses of N, P, and K. Both farms imported feed as well as nutrient inputs for crop production. Surpluses of all nutrients were typical in comparison to WFNBs of similar dairy farms; however, the nutrient use efficiencies were low. The imported manure and fertilizer used in the crop production components contributed to surpluses of N and P which could likely be reduced to improve overall farm nutrient use efficiency.

Glutamine (Gln) is a critical intermediate in nitrogen metabolism in all organisms. Here, a whole cell biosensor (GlnLux) for Gln was constructed by transforming a bacterial Gln auxotroph with a constitutive lux reporter. The biosensor was optimized for sensitivity, linearity, efficiency, specificity and robustness to permit detection of Gln in vitro and in vivo. The optimized GlnLux biosensor achieved nanomolar sensitivity with Gln standards. Extracts from only 1 mg of maize (Zea mays L.) leaf tissue were sufficient for Gln detection by GlnLux. Measurements of Gln in leaf extracts by GlnLux correlated with quantification by high performance liquid chromatography (Spearman r = 0.95). GlnLux permitted indirect in planta imaging of Gln using a CCD camera, enabling identification of plants that had been fertilized with nitrogen. Imaging using GlnLux also resolved predicted spatial differences in leaf Gln concentration. In a second application, it was demonstrated that GlnLux embedded into agar permits non-destructive screening of co-inoculated bacterial colonies for biological nitrogen fixation (BNF). GlnLux agar was able to distinguish a Bradyrhizobium japonicum wild type strain (nif+) from a mutant strain defective in nitrogenase (nif-) following ≥8 h of co-incubation. The technology was used to screen a bacterial endophyte diversity library cultured from Zea mays (L.) seeds for biological nitrogen fixation. / OMAFRA

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The soybean (Glycine max) cultivation in rotation with rice (Oryza sativa) in paddy soils is a growing practice in the state of Rio Grande do Sul. However, there is little information on the emission of methane (CH4) and nitrous oxide (N2O) during soybean growing season in lowland and the contribution of biological nitrogen fixation (BNF) for crops in this environment. The study was aimed to evaluate the emissions of CH4 and N2O, BNF and soybean productivity in lowland managed for three years with different systems of soil tillage/straw after the rice harvest. The experiment was conducted during the growing season of 2012/2013 on a Plano soil. The treatments consisted of different systems of soil management/straw applied for three years after the rice harvest: ryegrass (RY), disc harrow (DH), knife-roller (KR), fallow + disc harrow (F + DH), disc harrow + disc harrow (DH + DH), knife-roller + disc harrow (KR + DH), removal of straw (RS) and fallow (F). Besides these, a natural lowland (NL) was used as control. The NL had the lowest N2O emissions; however CH4-C (161 kg ha-1) emissions were higher when compared to soybean cultivated in lowland. The highest N2O emissions occurred from soybean cultivation, being observed in the treatment RY where the highest cumulative emission was 7.9 kg N2O-N ha-1. The management of soil/straw performed with DH, compared to systems with KR and F, caused increased density by up to 10% and reduction in macro-porosity and total porosity in the topsoil. Changes in soil physical properties caused by different soil/straw tillage systems did not result in a decreased N accumulation, BNF contribution (mean 67%) and productivity of soybean. / O cultivo da soja (Glycine Max) em rotação com o arroz (Oryza sativa) em solos de várzea é uma prática crescente no estado do Rio Grande do Sul (RS). No entanto, existem poucas informações sobre a emissão de metano (CH4) e óxido nitroso (N2O) durante o cultivo da soja em várzea e a contribuição da fixação biológica (FBN) para a cultura nesse ambiente. O estudo teve por objetivo avaliar as emissões de CH4 e N2O, a FBN e a produtividade da soja em várzea manejada durante três anos com diferentes sistemas de manejo do solo/palha após a colheita do arroz. O experimento foi conduzido no ano agrícola 2012/2013 em um Planossolo Hidromórfico Eutrófico arênico. Os tratamentos foram compostos por diferentes sistemas de manejo do solo/palha aplicados durante três anos após a colheita do arroz: azevém (AZ), grade (G), rolo-faca (RF), pousio + grade (P+G), grade + grade (G+G), rolo-faca + grade (RF+G), retirada da palha (RP) e pousio (P). Além desses, foi avaliada uma área natural de banhado (AN) que serviu como testemunha. A AN apresentou as menores emissões de N2O, porém elevada emissão de C-CH4 (161 kg ha-1) quando comparada ao cultivo da soja em várzea. As maiores emissões de N2O ocorreram no período de cultivo da soja, sendo observada no tratamento AZ a maior emissão acumulada desse gás (7,9 kg de N-N2O ha-1). O manejo do solo/palha realizado com G, comparado aos sistemas com RF e P, provocam aumento da densidade em até 10% e redução na macroporosidade e porosidade total na camada superficial do solo. As modificações causadas nos atributos físicos do solo pelos diferentes sistemas de manejo do solo/palha, não resultaram em diminuição no acúmulo de N, contribuição da FBN (média de 67%) e produtividade de grãos de soja.