Prediction and validation of gold nanoparticles (GNPs) on plant growth promoting rhizobacteria (PGPR): a step toward development of nano-biofertilizers

Several soil microbes are present in the rhizosphere zone, especially plant growth promoting rhizobacteria (PGPR), which are best known for their plant growth promoting activities. The present study reflects the effect of gold nanoparticles (GNPs) at various concentrations on the growth of PGPR. GNPs were synthesized chemically, by reduction of HAuCl 4, and further characterized by UV-Vis spectroscopy, X-ray diffraction technique (XRD), and transmission electron microscopy (TEM), etc. The impact of GNPs on PGPR was investigated by Clinical Laboratory Standards Institute (CLSI) recommended Broth-Microdilution technique against four selected PGPR viz., Pseudomonas fluorescens, Bacillus subtilis, Paenibacillus elgii, and Pseudomonas putida. Neither accelerating nor reducing impact was observed in P. putida due to GNPs. On the contrary, significant increase was observed in the case of P. fluorescens, P. elgii, and B. subtilis, and hence, GNPs can be exploited as nano-biofertilizers.

Avaliação técnica e econômica de um biodigestor de fluxo tubular: estudo de caso do modelo implantado na etec "orlando quagliato" em Santa Cruz do Rio Pardo, SP

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

Développement des marqueurs moléculaires spécifiques pour l'identification et la quantification de deux espèces de champignons mycorhiziens arbusculaires en utilisant la PCR en temps réel

Les champignons mycorhizien à arbuscules (CMA) sont des organismes pouvant établir des symbioses avec 80% des plantes terrestres. Les avantages d'une telle symbiose sont de plus en plus caractérisés et exploités en agriculture. Par contre, jusqu'à maintenant, il n'existe aucun outil permettant à la fois l'identification et la quantification de ces champignons dans le sol de façon fiable et rapide. Un tel outil permettrait, entre autres, de mieux comprendre les dynamiques des populations des endomycorhizes dans le sol. Pour les producteurs d'inoculum mycorhiziens, cela permettrait également d'établir un suivi de leurs produits en champs et d'avoir un contrôle de qualité de plus sur leurs inoculants. C'est ce que nous avons tenté de développer au sein du laboratoire du Dr. Hijri. Depuis environ une trentaine d'années, des outils d'identification et/ou de quantification ont été développés en utilisant les profiles d'acides gras, les isozymes, les anticorps et finalement l'ADN nucléaire. À ce jour, ces méthodes d’identification et de quantification sont soit coûteuses, soit imprécises. Qui plus est, aucune méthode ne permet à la fois la quantification et l’identification de souches particulières de CMA. L’ADN mitochondrial ne présente pas le même polymorphisme de séquence que celui qui rend l’ADN nucléaire impropre à la quantification. C'est pourquoi nous avons analysé les séquences d’ADN mitochondrial et sélectionné les régions caractéristiques de deux espèces de champignons mycorhiziens arbusculaires (CMA). C’est à partir de ces régions que nous avons développé des marqueurs moléculaires sous forme de sondes et d’amorces TaqMan permettant de quantifier le nombre de mitochondries de chacune de ces espèces dans un échantillon d’ADN. Nous avons ensuite tenté de déterminer une unité de quantification des CMA, soit un nombre de mitochondries par spore. C’est alors que nous avons réalisé que la méthode de préparation des échantillons de spores ainsi que la méthode d’extraction d’ADN avaient des effets significatifs sur l’unité de quantification de base. Nous avons donc optimisé ces protocoles, avant d’en e tester l’application sur des échantillons de sol et de racines ayant été inoculés avec chacune des deux espèces cibles. À ce stade, cet outil est toujours semi-quantificatif, mais il permet 9 l’identification précise de deux espèces de CMA compétentes dans des milieux saturés en phosphore inorganique. Ces résultats , en plus d’être prometteurs, ont permis d’augmenter les connaissances méthodologiques reliées à la quantification des CMA dans le sol, et suggèrent qu’à cause de leurs morphologies différentes, l’élaboration d’un protocole de quantification standardisé pour toutes les espèces de CMA demeure un objectif complexe, qui demande de nouvelles études in vivo.

Biofertilizantes: identificação das barreiras regulatórias e propostas para viabilizar esse insumo agrícola

O tema central dessa pesquisa é identificar gargalos da legislação brasileira por meio de revisão das principais leis, decretos e instruções normativas que norteiam o ambiente regulatório para os biofertilizantes, e propor medidas que facilitem o desenvolvimento do setor. O insumo faz parte da legislação brasileira desde a publicação da primeira lei de fertilizantes em 1980, e tem seu uso descrito em trabalhos acadêmicos. No entanto, a falta de registros de produtos biofertilizantes junto ao Ministério da Agricultura, Pecuária e Abastecimento (MAPA) até o final do ano de 2014 merece atenção do setor de insumos e das lideranças que legislam. O entendimento das implicações e efeitos da legislação na adoção do insumo, bem como as implicações para a revisão das normas, presentes no trabalho, apoiam o setor de insumos, as lideranças políticas e a pesquisa na adequação das normas e procedimentos técnicos que viabilizem o registro das soluções do setor de biofertilizantes e o acesso facilitado por parte dos agricultores. O trabalho revisou a legislação brasileira relevante para o insumo biofertilizante, e entrevistou lideranças políticas, institucionais e de empresas no que diz respeito às principais dificuldades enfrentadas para o melhor desenvolvimento do setor. Investigou-se também a situação apresentada em outros países, como Estados Unidos, Canadá e países da União Européia através da revisão de leis e documentos, e de entrevistas com profissionais do setor de insumos e representantes de instituições atuantes nos mercados mencionados. Apesar de diferenças nas definições propostas pelos vários países e outras classificações congruentes, as dificuldades que empresas do setor de nutrição enfrentam no registro de biofertilizantes correspondem aos desafios também enfrentados por empresas para criar a classe de bioestimulantes nos Estados Unidos, e principalmente àqueles da maioria dos países europeus analisados. Identificou-se que a legislação vigente e sua interpretação, e exigências para o lançamento de novos produtos não atendem plenamente o setor produtor de biofertilizantes, o qual ademais não possui a mesma expertise regulatória que o setor agroquímico para melhor definir seus protocolos de validação junto ao órgão anuente. Mudanças na legislação, melhor articulação da cadeia produtiva no entendimento das necessidades, e maior interação das entidades representativas com o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) se fazem essenciais para viabilidade do insumo biofertilizante.

Interação de microorganismos na solubilização de fósforo e potássio de rochas para produção de biofertilizantes

he present model of agriculture is based on intensive use of industrial inputs, due to its rapid response, but it brings harmful consequences to the environment, and it is necessary the use of modern inputs. And an alternative is the use of rock biofertilizers in agriculture, a product easy to use, with higher residual effect and does not harm the environment. The objective of study was to evaluate the inoculation and co-inoculation of different microorganisms in the solubilization of rock phosphate and potash ground microbial evaluating the best performance in the production of biofertilizers comparing with rocks pure in soil chemical properties and, verify effect of inoculation of the bacterium Paenibacillus polymyxa in the absorption of minerals dissolved in the development of cowpea (Vigna unguiculata [L.] Walp.). The first bioassay was conducted in Laboratory (UFRN) for 72 days in Petri dishes, where the rock powder was increased by 10% and sulfur co-inoculated and inoculated with bacterial suspension of Paenibacillus polymyxa grown in medium tryptone soy broth, Ralstonia solanacearum in medium Kelman, Cromobacterium violaceum in medium Luria-Bertani and Acidithiobacillus thiooxidans in medium Tuovinen and Kelly,and fungi Trichoderma humatum and Penicillium fellutanum in malt extract. Every 12 days, samples were removed in order to build up the release curve of minerals. The second bioassay was conducted in a greenhouse of the Agricultural Research Corporation of Rio Grande do Norte in experimental delineation in randomized block designs, was used 10 kg of an Yellow Argissolo Dystrophic per pot with the addition of treatments super phosphate simple (SS), potassium chloride (KCl), pure rock, biofertilizers in doses 40, 70, 100 and 200% of the recommendation for SS and KCl, and a control, or not inoculated with bacteria P. polymyxa. Were used seeds of cowpea BRS Potiguar and co-inoculated with the bacterial suspension of Bradyrhizobium japonicum and P. polymyxa. The first crop was harvested 45 days after planting, were evaluated in the dry matter (ADM), macronutrients (N, P, K, Ca, Mg) and micronutrients (Zn, Fe, Mn) in ADM. And the second at 75 days assessing levels of macro end micronutrients in plants and soil, and the maximum adsorption capacity of P in soil. The results showed synergism in co-inoculations with P. polymyxa+R. solanacearum and, P. polymyxa+C. violaceum solubilizations providing higher P and K, respectively, and better solubilization time at 36 days. The pH was lower in biofertilizers higher doses, but there was better with their addition to P at the highest dose. Significant reduction of maximum adsorption capacity of phosphorus with increasing dose of biofertilizer. For K and Ca was better with SS+KCl, and Mg to pure rock. There was an effect of fertilization on the absorption, with better results for P, K and ADM with SS+KCL, and N, Ca and Mg for biofertilizers. Generally, the P. polymyxa not influence the absorption of the elements in the plant. In treatments with the uninoculated P. polymyxa chemical fertilizer had an average significantly higher for weight and number of grains. And in the presence of the bacteria, biofertilizers and chemical fertilizers had positive values in relation to rock and control. The data show that the rocks and biofertilizers could meet the need of nutrients the plants revealed as potential for sustainable agriculture

Biofertilizer for control of Guignardia citricarpa, the causal agent of citrus black spot

Biofertilizers generated from the anaerobic and aerobic digestion of cattle manure, known as Bio1 and Bio2, respectively, were studied with regard to their microbial composition and effect on the mycelial growth of Phyllosticta citricarpa, the causal agent of citrus black spot (CBS). Two field experiments were conducted to determine the biofertilizer's potential (Bio1) in controlling CBS (2001/2002 and 2002/2003 crops). It was observed that the greatest number of microorganisms was found in the aerobically produced biofertilizer. Bio2 did not inhibit the mycelial growth of P. citricarpa. Mycelial growth of P. citricarpa was inversely proportional to the Bio1 biofertilizer concentration. In the 2001/2002 cropping season, the Biol effect in controlling CBS was directly proportional to its concentration, at the rate of 0 (healthy fruit), with R-2 = 0.88. Biol had a significant effect in controlling CBS, at a concentration of 10%, during the 2001/2002 cropping season, with DI values of 0.246 and 0.229 for the. 10 and 20% doses, respectively, compared to DI of 0.329 for the control. A directly proportional effect of the biofertilizer concentration on the percentage of fruits with a rating of zero was. also observed in the 2002/2003 cropping season, with R-2 = 0.48. However, even at doses higher than in the preceding cropping season, the biofertilizer was less effective, possibly due to a higher occurrence of the disease. Copper oxychloride and combined applications of copper oxychloride and carbendazim plus mancozeb controlled the disease. The possibility of using the biofertilizer as a protective biofungicide to replace copper oxychloride, especially in organic agriculture, should be explored. (c) 2005 Elsevier Ltd. All rights reserved.

Controle alternativo de Colletotrichum acutatum agente causal da queda prematura dos frutos cítricos

A queda prematura dos frutos cítricos (QPFC), causada por Colletotrichum acutatum, dados os grandes prejuízos que têm causado aos produtores, constitui-se numa doença de grande importância econômica. O controle da doença é feito predominantemente mediante uso de fungicidas, que eleva o custo de produção e afeta negativamente o meio ambiente. Diante disso, este trabalho teve por objetivo buscar um método alternativo de controle da QPFC, mediante o uso de agentes de biocontrole ou de biofertilizantes. Diferentes concentrações de biofertilizantes (originários de duas fontes distintas e denominados de Bio1 e Bio 2); três isolados de Bacillus subtilis (ACB-69; 72 e 77) e três isolados de Trichoderma spp. (ACB-14; 37 e 39) foram testados, isoladamente ou em combinação, sob condições de laboratório, quanto à capacidade inibitória da germinação de conídios de C. acutatum. Estudaram-se, ainda, a produção de metabólitos termoestáveis por B. subtilis e o efeito sobre a germinação do patógeno. Quinze isolados de B. subtilis foram testados quanto à capacidade de prevenir a infecção por C. acutatum em flores destacadas de lima- ácida 'Tahiti' e, no campo, foram instalados dois experimentos, visando a testar ACBs e biofertilizantes no controle da doença. Verificou-se que o isolado ACB-72 (B. subtilis) e ACB-37 (T. pseudokoningii) foram os que mais inibiram a germinação do patógeno. Quanto à produção de metabólitos termoestáveis, ACB-69 e 77 foram os mais eficientes em produzir substâncias antifúngicas, e em quantidades suficientes para inibirem a germinação do patógeno. A mistura dos quatro isolados de Bacillus (ACBs: 69; 72; 77 e AP3) foi o que apresentou maior porcentagem de inibição (73%). Os biofertilizantes (Bio1 e Bio2), em concentrações acima de 10% e, quando em associação com isolados de Trichoderma spp., promoveram maiores inibições na germinação de C. acutatum. em testes com flores destacadas, verificou-se que, onde foram aplicados os ACBs 69; 76; 74 e 77, as porcentagens de pétalas sem sintomas de infecção por C. acutatum foram de 83; 92; 92 e 97%, respectivamente. Mediante avaliações a campo, verificou-se a potencialidade de B. subtilis e de biofertilizantes em controlar a doença.

Aclimatização de mudas micropropagadas de bananeira 'Grand Naine' com aplicação de biofertilizantes em duas estações do ano

A utilização de mudas de bananeira micropropagadas de qualidade é o primeiro passo na implantação de um bom pomar e, para isso, sua aclimatização deve ser realizada de forma adequada. em vista disso, objetivou-se, neste trabalho, avaliar o desempenho dos biofertilizantes HUMITEC® e RUTER AA® no desenvolvimento de mudas de bananeira micropropagadas, sendo instalado um experimento em viveiro comercial de produção de mudas. Foram utilizadas mudas de bananeira cv. Grand Naine, obtidas por micropropagação, sendo, posteriormente, transplantadas em sacos de polietileno preto (1,5 L), contendo, como substrato, terra de subsolo (Latossolo Amarelo), casca de arroz carbonizada e composto orgânico Organifol® (1:1:1). O delineamento experimental foi em blocos ao acaso, fatorial 2 x 4 (produtos e doses), com quatro repetições e quatro plantas úteis por parcela. Os tratamentos foram compostos pelos seguintes biofertilizantes e doses: HUMITEC® (0,0; 4,0; 8,0; 16,0 mL planta-1) e RUTER AA® (0,0; 2,0; 4,0; 8,0 mL planta-1). As doses dos produtos foram divididas em duas aplicações (28 e 56 dias após o transplantio das mudas), aplicadas via fertirrigação. Noventa e oito dias após o transplantio avaliaram-se as seguintes variáveis: altura das plantas (cm), diâmetro do colo rente superfície do solo (mm), área foliar total e por folha, biomassas fresca e seca da parte aérea e das raízes (g). Os dados foram submetidos análise da variância e quando o teste F foi significativo, realizou-se análise de regressão. Pode-se concluir que aplicações de HUMITEC® e RUTER AA® favoreceram o desenvolvimento das mudas e que, no verão, as plantas apresentaram maior desenvolvimento durante a aclimatização.

Produtividade e qualidade de frutos do mamoeiro em função de tipos e doses de biofertilizantes

Biofertilizers are organic products usually obtained from an anaerobic process, evaluated mainly as phytoprotectors for agriculture, although they can improve fruit production. By this way, a field experiment was carried out in Remigio County, Paraiba State, Brazil, to evaluate the effects of two bovine biofertilizers on yield, medium mass and quality of papaya fruits (Carica papaya L.), 'Baixinho de Santa Amalia cultivar. Treatments were distributed in a randomized complete blocks design with three replications and 15 plants in each parcel, in a 2 x 5 factorial arrangement, with two biofertilizers: simple (water and fresh bovine manure) and supermagro (water, fresh bovine manure, macro and micronutrients and a protein mix), applied in soil under liquid form at five volumes, as follows: 0.0; 0.5; 1.0; 1.5; e 2.0 L cave(-1). There was no statistical difference between biofertilizers for fruit mass, yield and external and internal fruit quality variables, while between volumes differences were registered for both biofertilizers. The results, except for fruit firmness, registered positive effects of the biofertilizers on 'Baixinho de Santa Amalia' papaya cultivar in relation to yield and physical and chemical fruit quality.

Effects of biofertilizer rates on the structural, morphogenetic and productive characteristics of Piata grass

The objective of this study was to investigate the influence of different levels of biofertilizers from cattle and swine manure on the structural, morphogenetic and productive characteristics of Brachiaria brizantha cv. Piata. The experiment was arranged in a completely randomized factorial design with split plots. The plots were defined by eight treatments: two biofertilizers (cattle and swine), four levels (0, 100, 200 and 300 kg N.ha(-1)) and subplots by four different cutting periods. The cutting for plant uniformity was performed at 45 days after sowing at 15 cm above the soil surface. The biofertilizeres were applied in a single level, after the cutting of plants, in rates of 0, 0.23 and 0.19, 0.45 and 0.38, 0.68 and 0.57 liters pot(-1) for the biofertilizers from cattle and swine manure, respectively. These rates were also equivalent to levels of 0, 100, 200 and 300 kg N.ha(-1). There was no significant difference between the types of biofertilizers as there was no interaction between them and the different levels, hence both biofertilizers could be applied without any loss of nutrient intake by the plants used in this experiment. There was a significant difference between the production of green and dry matter, the leaf appearance rate, phyllochron, leaf and pseudostem elongation rates, number of green leaves, final leaf length, number and weight of tillers, according to the increase of nitrogen rates, following linear prediction model. Effect of the cutting periods was also observed, once the plants harvested during the summer presented greater performance of structural and morphogenetic characteristics.

Adubação organomineral e biofertilização líquida na produção de frutos de pinheira ( Annona squamosa L.) no submédio São Francisco, Brasil

The organic production systems based on natural processes do not allow the use of synthetic fertilizers with a high solubility, so it is required the use of technologies in accordance with the organic production legislation in order to make the technical, economical and environmental sustainable activities viable. To evaluate the effects of the usage of mineral and organic natural fertilizers, associated with liquid biofertilizers (BLE= enriched liquid biofertilizer and BF= leaf biofertilizers) on the production of the sugar apple fruit crop Annona squamosa L., replacing totally the chemical fertilizers, used by conventional farming systems, a nine year old orchard was driven in the irrigated plot n degrees 1295, irrigated project Nilo Coelho, district of Petrolina - PE. The experiment was carried out from September 2005 to February 2006. The experimental design was done through randomized blocks with eight treatments and four replications in plots of three plants each, with 96 plants in total. The treatments were the following: T(1) - plants with no fertilizer; T(2) - 60 g de N + 32 g de P(2)O(5) + 48 g de K(2)O; T(3) - 90g de N + 32 g de P(2)O(5) + 48 g de K(2)O; T(4) - 30 g de N + 32 g de P(2)O(5) + 48 g de K(2)O + 15 L de BLE + BF a 5 %; T(5) - 60g de N + 32 g de P(2)O(5) + 48 g de K(2)O + 30 L de BLE + BF a 5 %; T(6)- 90g de N + 32 g de P(2)O(5) + 48 g de K(2)O + 45 L de BLE + BF a 5 %; T(7) - 120g de N + 32 g de P(2)O(5) + 48 g de K(2)O + 60 L de BLE + BF a 5 %; T(8) - 150 g de N + 32 g de P(2)O(5) + 48 g de K(2)O + 75 L de BLE + BF a 5 % The fruit growth analyses (diameter and average length), total yield, productivity, average fruit weight, average number of fruits, fresh weight of the skin and pulp and dry weight of the fruits were used to evaluate the experiment. The results were relevant to the total production, fresh weight of the skin and dry weight of the fruits. The use of organominerals associated with liquid fertilizers increased significantly the productivity in values ranging from 10.44% (T(5) = 1.98 t ha(-1)) to 24.52 % (T(7) = 12.34 %). The treatment T(3) stood out (90g de N + 32g de P(2)O(5) + 48g de K(2)O) with a higher production than the control. The fruit weight increased in the fertilized treatments, with values increasing from treatment (T(2) = 5.26 %) to the treatment (T(7) = 12.34 %) in relation to the control. The fruit development was characterized by an average growth increase of 82 % in length and 84 % in diameter until the 56(th) day, with a sigmoidal growth pattern.

Efeito de biofertilizantes sobre o crescimento micelial e a germinação de esporos de alguns fungos fitopatogênicos

The biofertilizer was produced through anaerobic fermentation of cow manure adding milk, sugar, salts, cow liver parts and bone powder. After 73 days of fermentation it was evaluated the effect on micelial growth of Pythium aphanidermatum, Alternaria solani, Stemphylium solani, Septoria licopersici, Sclerotinia sclerotiorum, Botrytis cinerea, Rhizoctonia solani, Fusarium oxysporum f. sp. phaseoli and spores germination of B. cinerea, A. solani, Hemileia vastatrix and Coleosporium plumierae. In relation to micelial growth inhibition, the growth rate was calculated and it was found that, in general, concentrations over 10% caused a total inhibition of growth for the majority of fungi assayed. In case of spores germination, biofertilizer concentration over 20% has inhibited completely the germination of B. cinerea, over 10% inhibited A. solani, 5 and 1% of C. plumierae and H. vastatrix, respectively. Three different biofertilizers were also tested and one of them was less effective, which was the one produced with manure from confined cows opposed to the others produced with grazing cows.

Resposta da alface à aplicação de biofertilizante sob dois níveis de irrigação

The use of biofertilizers is interesting for agriculture as being an economical alternative as well as it is environmentally friendly by using organic waste and reducing the application of mineral fertilizers. The aim of this study was to evaluate the effect of biofertilizer doses of bovine origin (biodigester effluent) applied on the ground and two levels of irrigation on lettuce. The experiment was conducted under protection of a greenhouse in pots, applying to the soil different doses of biofertilizer of bovine origin obtained from anaerobic reactor (10, 20, 40 and 60 m3 ha-1) and mineral fertilizer as a witness in two irrigation levels calculated at 50 and 100% of reference evapotranspiration. The lettuce plants were analyzed in their: Height, leaves number, crown diameter, fresh weight and dry weight of shoots. The biofertilizer treatments showed better results than the mineral fertilizer and has increased with increasing doses of biofertilizer, the highest dose (60 m3 ha-1) showed the best results in all variables. For dry, mineral fertilization showed higher values. The irrigation levels had no effect on plant growth.

Biofertilização na produção de alfafa e atributos

Pós-graduação em Ciência Animal - FMVA

Estudo do potencial do licor pirolenhoso como aditivo em condicionador de solo e no crescimento inicial de eucalipto

Pós-graduação em Agronomia (Energia na Agricultura) - FCA