Toxicity of Herbicide, Paraquat Dichloride and Insecticide, Lambda-cyhalothrin on Phosphate Solubilizing Bacteria, Pantoa dispersa in Aquatic Ecosystems

Aim: The study was aimed to determine the toxicity of paraquat dichloride and lambda-cyhalothrin on phosphate solubilizing bacteria, Pantoa dispersa in aquatic ecosystems.Study Design: This study employs randomized block design, statistical analysis of the data and interpretation.Place and Duration of the Study: Soil sample was collected from the root nodules of leguminous plants in a sterile polythene bag from the Elele, in Etche L.G.A, Rivers State. The fresh water sample was collected from Bane town in Khana L.G.A, brackish water sample was collected from Choba river in Obio/Akpor L.G.A while the marine water was collected from Bonny River of Bonny L.G.A., all of Rivers State, Nigeria. The samples were collected aseptically and transported in an ice-pack immediately to the Rivers State University, Microbiology laboratory for analysis. The study lasted for three months.Methodology: The bacterium, Pantoa dispersa was isolated and identified based on conventional and molecular characterization from water and soil samples. Different concentrations (3.13%, 6.25, 12.50, 25.00%, 50.00% and 75.00%) and the control (0%) of the herbicide (paraquat dichloride) and insecticide, Lambda-cyhalothrin were prepared using fresh, brackish and marine water samples and 10ml of the test organism, Pantoa dispersa was introduced and the survival count was determined at 0, 4hr, 8hr, 12hr and 24hr. The Mean Lethal Concentration (LC50) of the insecticide and herbicide on Pantoa dispersa in the three aquatic ecosystem was determined.Results: The LC50 of the herbicide (Paraquat dichloride) was recorded as 15.8% in brackish water, 17.37% in fresh water and 27.44% in marine water. While the LC50 of the insecticide, Lambda-cyhalothrin to Pantoa dispersa was 26.84% in fresh water, 27.26% in brackish water and 32.33% in marine water.Conclusion: From the study, the herbicide, Paraquat dichloride was more toxic in the three aquatic ecosystems compared to the insecticide, Lambda-cyhalothrin. The use of these agrochemicals should be monitored as they result in the mortality of beneficial soil bacteria like Pantoa dispersa which is phosphate solubilizing bacteria in aquatic ecosystems.

Optimization of soybean yield in ultisols through adaptive varieties screening and plant growth promoting Rhizobacter

This research was aimed at obtaining varieties of soybean adaptive to acid soils and to obtain Plant Growth-Promoting Rhizobacteria (PGPR) isolates that can improve the agronomic characteristics of soybean and increase the ultisols fertility. This research was conducted in two-stages research on Sampali Village, Percut Sei Tuan sub-District, Deli Serdang District, North Sumatra Province, Indonesia from August 2019 until March 2020. The first stage (adaptive varieties screening) using the non-factorial Randomized Block Design (RBD) with the varieties of Argomulyo, Wilis, Kaba, Dena-1, Devon-1, Dega-1, Demas-1, Burangrang, Detam-1, and Kipas Merah. The second stage (application of PGPR isolates singly and in combinations) using the factorial RBD, the first factor of applicative single and the combination of PGPR isolates, the second factor of adaptive varieties including Detam-1 and Wilis. Data were analyzed with ANOVA and followed by DMRT at P<0.05. The results showed that the Detam-1 and Wilis varieties had significantly higher yield per plant of 14.73 g and 14.54 g, respectively, than other varieties. The applications of a single and combination of PGPR isolates significantly increased the number of branches, stem diameter, plant height, yield per plant, soil pH, organic-C, available-P, and total-N and decreased the soil C/N. The Detam-1 variety had the higher in yield per plant compared to Wilis variety. The isolates combination of Rhizobium leguminosarum+Rhizobium sp2+Bacillus sp+Burkholderia sp for Detam-1 and Wilis varieties can be recommended to support the growth and yield of soybean on ultisols.

Agronomic efficiency and profitability of cotton on integrated use of phosphorus and plant microbes / Eficiência e rentabilidade agronômica do algodão no uso integrado de fósforo e micróbios vegetais

Cotton crop, plays a significant role in Pakistan's economy by ruling a prominent place in edible oil and local textile industry. Phosphorus (P) inaccessibility and deficiency of soil organic matter are the key restraints for low crop productivity in cotton. Therefore, a two years field study was designed during 2014-15, to explore the influence of phosphate solubilizing bacteria (PSB), farmyard manure (FYM), poultry manure (PM) and inanimate sources of P on various physiological, growth, yield and quality parameters of cotton crop at CCRI Multan. Field responses of seeds inoculated with two distinctive phosphate solubilizing bacteria (PSB) strains viz. S0 = control, S1 =strain-1, S2 = strain-2 and eight organic, inorganic P sources viz., P0= control, P1 = 80 kg ha-1 P from inorganic source, P2 = 80 kg ha-1 P from FYM, P3 = 80 kg ha-1 P from PM, P4 = 40 kg ha-1 P from FYM + 40 kg ha-1 P from inorganic source, P5 = 40 kg ha-1 P from PM + 40 kg ha-1 P from inorganic source, P6 = 80 kg ha-1 P from FYM + 40 kg ha-1 P from inorganic source, P7 = 80 kg ha-1 P from PM + 40 kg ha-1 P from inorganic source and P8 = 40 kg ha-1 P from FYM + 40 kg ha-1 P from PM were evaluated. Results revealed that inoculation of seeds with PSB and collective use of inorganic and organic sources of P had considerably increased the yield contributing attributes in cotton. However, the treatment P7 (80 kg P ha-1 from PM + 40 kg P ha-1 from inorganic source) in coincidence with seeds inoculated with PSB (S1) produced taller plant, maximum boll weight, significantly higher LAI and CGR. Significantly higher seed cotton yield, lint yield, fiber length and maximum BCR of 1.95 and 1.81 was also obtained from the P7 treatment during both crop-growing seasons. In conclusion, combined use of 80 kg P ha-1 from PM + 40 kg P ha-1 from inorganic source and cotton seeds inoculated with strain-1 improved phosphorus uptake ensuing in greater consumption of photo-assimilates for maximum growth and yield.

A safra de algodão, desempenha um papel significativo na economia do Paquistão, ao ocupar um lugar de destaque no óleo comestível e na indústria têxtil local. A inacessibilidade e a deficiência de fósforo (P) da matéria orgânica do solo são as principais restrições para a baixa produtividade das culturas em algodão. Portanto, um estudo de campo de dois anos foi desenvolvido durante 2014-15, para explorar a influência de bactérias solubilizantes de fosfato (PSB), esterco de capoeira (FYM), esterco de aves (PM) e fontes inanimadas de P sobre vários fatores fisiológicos, crescimento, rendimento e parâmetros de qualidade da cultura do algodão no CCRI Multan. Respostas de campo de sementes inoculadas com duas cepas distintas de bactérias solubilizantes de fosfato (PSB) viz. S0 = controle, S1 = cepa-1, S2 = cepa-2 e oito fontes orgânicas de P inorgânicas viz., P0 = controle, P1 = 80 kg ha-1 P de fonte inorgânica, P2 = 80 kg ha-1 P de FYM, P3 = 80 kg ha-1 P de PM, P4 = 40 kg ha-1 P de FYM + 40 kg ha-1 P de fonte inorgânica, P5 = 40 kg ha-1 P de PM + 40 kg ha-1 P de fonte inorgânica, P6 = 80 kg ha-1 P de FYM + 40 kg ha-1 P de fonte inorgânica, P7 = 80 kg ha-1 P de PM + 40 kg ha-1 P de fonte inorgânica e P8 = 40 Foram avaliados kg ha-1 P da FYM + 40 kg ha-1 P da PM. Os resultados revelaram que a inoculação de sementes com PSB e o uso coletivo de fontes inorgânicas e orgânicas de P aumentaram consideravelmente os atributos que contribuem para a produtividade no algodão. No entanto, o tratamento P7 (80 kg P ha-1 da PM + 40 kg P ha-1 da fonte inorgânica) em coincidência com sementes inoculadas com PSB (S1) produziu planta mais alta, peso máximo de cápsula, IAF e CGR significativamente maiores. Produtividade significativamente maior do algodão, rendimento de fiapos, comprimento da fibra e BCR máximo de 1,95 e 1,81 também foi obtida a partir do tratamento P7 durante as duas épocas de cultivo. Em conclusão, o uso combinado de 80 kg P ha-1 de PM + 40 kg P ha-1 de fonte inorgânica e sementes de algodão inoculadas com a cepa-1 melhoraram a captação de fósforo, resultando em maior consumo de foto assimilados para obter crescimento e produtividade máximos.

Isolation and identification of phosphate solubilizing bacteria from indigenous microorganisms (IMO) of cow rumen in East Java, Indonesia as eco-friendly biofertilizer

Aims@#Phosphate is an essential nutrient required for plant growth, but its solubility in the soil is relatively low (0.1%). Microbes can dissolve phosphate to meet crop requirements. This study aimed to isolate phosphate solubilizing bacteria from indigenous microorganisms (IMO) of cow rumen.@*Methodology and results@#The selection of isolates on a Pikovskaya medium was using a clear zone index and a spectrophotometer for phosphate solubilization measurements. Hypersensitivity was tested on tobacco leaves and tested antagonists within isolates. The results found that four selected isolates had the highest phosphate dissolving potential, namely, MTA1, SMAD1, SMAD2, and SMAD3. The culture of selected isolates on plate media showed that the morphological characters of the four colonies are the same. They had round form (circular), the edge of the colony were smooth, flat elevation, white and cream color. Isolate MTA1 had the highest phosphate solubilizing activity compared to the others. The isolate that showed the highest phosphate solubilizing activity were identified based on 16S rRNA gene. The result of molecular identification showed that strain MTA1 was closely related to Lactobacillus plantarum with a similarity level of 99%. L. plantarum performed the highest ability to form a clear zone (7.66 mm). The highest concentration of soluble phosphate was observed on day 5 (278.42 mg/L).@*Conclusion, significance, and impact of the study@#Lactobacillus plantarum which was isolated from the IMO of cow rumen in East Java, Indonesia was identified as one of the phosphate solubilizing bacteria that are useful for the development of eco-friendly biofertilizer. The application of phosphate solubilizing microbes can be used to increase the soil fertility.

Effect of management (ecological and conventional) on functional groups of soil microorganisms in coffee agroecosystems with different resilience to climate variability, Colombia

The effect of management (ecological and conventional) on functional groups of microorganisms of soil in agroecosystems with different resilience scores reported to climate variability in Anolaima, Colombia was evaluated. Were found clustering associated with management and cellulolytic bacteria and fungi abundances. No differences found in diversity of phosphate solubilizing or nitrogen-fixing microorganisms, related to management. The diversity of microbial functional groups was affected by the climatic condition of sampling season. Management was relevant in relationships between resilience scores to climate variability and cellulolytic microorganisms; in ecological agroecosystems, biodiversity knowledge, agroecological main structure, and the participation of farmers in organizations were important.

Phosphate-solubilizing function evaluation and taxonomic identification of endophytic fungi separated from roots of Amomum villosum / 中草药

Objective: To study the characteristics of endophytic fungi separated from the roots of Amomum villosum grown in Jinping County of Yunnan Province, including the culture, evaluation of phosphorus-solubilizing ability and taxonomic identification of target strains. Methods: Endophytic fungi in the roots of A. villosum were separated by culturing in the mediums of PDA and MEA, and purely cultured in PDA. The endophytic fungi with phosphorus-solubilizing ability were screened by solid and liquid mediums of Pikovaskaia’s (PVK) prepared with inorganic phosphorus source. Then, the phosphorus-solubilizing capacity and reasonable mechanism were analyzed by growth circle, biomass, effective phosphorus content, pH value, and phosphatase activity. Moreover, molecular identification of target strains with the capacity of phosphorus-solubilizing would be carried out by ribosome 18 S PCR amplification. Results: The results showed that 24 endophytic fungi were separated from the roots of A. villosum in total, 10 of which were dark septate endophytes (DSE). Eight strains could grow on PVK solid medium and produce phosporus-dissolved growth circle. The growth circle diameters of JP-20 and JP-23 were larger than others, and more than 9 cm, followed by JP-15 with the growth circle of 6.06 cm. Furthermore, it was shown that JP-23 had a strong ability of phosphate-solubilizing due to presenting a high biomass in the PVK liquid medium instead of the medium prepared by soluble phosphorus source. The content of effective phosphorus of JP-23 in PVK liquid medium was significantly increased with an obviously decreasing pH and a sharply rising of acid phosphatase (ACP) activity. Moreover, the strain JP-23 was preliminarily identified as Cladosporium sp. (GenBank: MK629004) by molecular identification. Conclusion: An assumption was concluded that strain JP-23 could decompose and use insoluble phosphorus sources by adjusting the pH value and secreting ACP in medium. Our findings would provide data support and theoretical basis for studying the phosphate absorption mechanism of plant-microbial symbiosis system and the ecological plantation of A. villosum.

Impact of phosphate solubilizing bacteria on wheat (Triticum aestivum ) in the presence of pesticides / Impacto das bactérias solubilizantes de fosfato no trigo (Triticum aestivum ) na presença de pesticidas

Abstract Three phosphate solubilizing bacteria were isolated and identified by 16S rRNA sequencing as Pseudomonas putida, Pseudomonas sp and Pseudomonas fulva . The strains were subjected to plant biochemical testing and all the PGPR attributes were checked in the presence of pesticides (chlorpyrifos and pyriproxyfen). The phosphate solubilizing index of strain Ros2 was highest in NBRIP medium i.e 2.23 mm. All the strains showed acidic pH (ranges from 2.5-5) on both medium i.e PVK and NBRIP. Strain Ros2 was highly positive for ammonia production as well as siderophore production while strain Rad2 was positive for HCN production. The results obtained by the strains Rad1, Rad2 and Ros2 for auxin production were 33.1, 30.67 and 15.38 µg ml-1, respectively. Strain Rad1 showed 16% increase in percentage germination in comparison to control in the presence of pesticide stress. Most promising results for chlorophyll content estimation were obtained in the presence of carotenoids upto 6 mgg-1 without stress by both strains Rad1 and Rad2. Study suggests that especially strain Ros2 can enhance plant growth parameters in the pesticide stress.

Resumo Três bactérias solubilizantes de fosfato foram isoladas e identificadas por seqüenciamento de rRNA 16S como Pseudomonas putida, Pseudomonas sp e Pseudomonas fulva. As estirpes foram submetidas a testes bioquímicos de plantas e todos os atributos PGPR foram verificados na presença de pesticidas (clorpirifos e piriproxifeno). O índice de solubilização de fosfato da estirpe Ros2 foi mais elevado no meio NBRIP, isto é, 2,23 mm. Todas as estirpes apresentaram um pH ácido (varia de 2,5-5) em ambos os meios, isto é PVK e NBRIP. A estirpe Ros2 foi altamente positiva para a produção de amoníaco, bem como a produção de sideróforos enquanto a estirpe Rad2 foi positiva para a produção de HCN. Os resultados obtidos pelas estirpes Rad1, Rad2 e Ros2 para a produção de auxina foram 33,1, 30,67 e 15,38 μg ml-1 , respectivamente. A deformação Rad1 mostrou aumento de 16% na germinação percentual em comparação com o controlo na presença de stress de pesticida. Os resultados mais promissores para a estimativa do teor de clorofila foram obtidos na presença de carotenóides até 6 mgg-1 sem estresse por ambas as cepas Rad1 e Rad2. Estudo sugere que especialmente a estirpe Ros2 pode melhorar parâmetros de crescimento de plantas no estresse de pesticidas.

Capacidad solubilizadora de fosfato de aluminio por hongos rizosféricos aislados de un Andisol colombiano / Solubilizing capacity of aluminum phosphate by rhizospheric fungi isolated from an Andisol Colombian / Capacidade de solubilização de fosfato de alumínio por fungos rizosféricos isolados de um Andisol colombiano

RESUMEN A partir de un Typic Melanudands cultivado con café (Coffea arabica variedad Caturra), proveniente del Departamento del Cauca (Colombia), se aislaron hongos con capacidad solubilizadora de fosfato de aluminio (Al-P), de los cuales se seleccionaron dos microorganismos que presentaron la mayor actividad, identificados como cepa UNHI (Mycelia sterilia), y UNH2 (Penicillium sp). Los hongos seleccionados fueron evaluados por quince días en medio líquido Pikovskaya (PVK) con Al-P bajo dos condiciones, con agitación (método A) y en reposo con aireación (método B). Al comparar los resultados se encontraron diferencias estadísticas significativas en el porcentaje de fósforo soluble, el método A presentó mejores resultados con un porcentaje de 73,8% frente a 62% con la cepa UNH1 y en menor proporción con la cepa UNH2 49% y 44%. El incremento de biomasa fue mayor con el método B y el pH de los medios de cultivo no mostraron diferencias significativas, con un pH promedio de 2,7.

ABSTRACT Based on Typic Melanudands grow with coffea (Coffea Arabica Caturra variety) resulting from the Cauca department, fungi with solubilization capacity of aluminum phosphate were isolated (Al-P), from which two microorganisms with greater solubilization activity were selected, identified as strain UNH!: Mycelia sterilia y UNH2: Penicillium sp. Selected fungi were evaluated through the Pikovskaya liquid (PVK) with (Al-P), under two conditions, with stirring (A method) and repose with aireation ( B method), for fifteen days. When comparing the results, significant statistical differences were found in the percentage of soluble phosphorus, method A presented better results with a percentage of 73.8% compared to 62% with the strain UNH1 and in smaller proportion with the strain UNH2 49% and 44 %. The increase in biomass was greater with method B and the pH of the culture media did not show significant differences, with an average pH of 2.7.

RESUMO De PVAc Melanudands cultivadas com café (Coffea arabica variedade caturra), a partir do Departamento (Colômbia), fungos foram isoladas com fosfato de alumínio capacidade de solubilização (Al-P), de que introduziu foram seleccionados dois microrganismos a atividade mais alta, identificada como cepa UNH1 (Mycelia sterilia) e UNH2 (Penicillium sp). fungos seleccionados foram avaliadas durante duas semanas em meio líquido Pikovskaya (PVK) com Al-P sob duas condições, com agitação (método A) e que descansa com arejamento (método B). Ao comparar os resultados estatisticamente diferenças significativas na percentagem de fósforo solúvel encontrada, método A proporcionou melhores resultados com uma percentagem de 73,8% versus 62% com a estirpe UNH1 e em menor grau com a estirpe UNH2 49% e 44 %. O aumento da biomassa foi maior com o método B e o pH do meio de cultura não mostrou diferenças significativas, com um pH médio de 2,7.

Endophytic Bacillus strains enhance pearl millet growth and nutrient uptake under low-P

Abstract Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.

Endophytic Bacillus strains enhance pearl millet growth and nutrient uptake under low-P

Abstract Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.

Identificación de bacterias solubilizadoras de fosfatos en un Andisol de la región cafetera colombiana / Identification of phosphate solubilizing bacteria in a Andisol of Colombian coffee region

Resumen El fósforo es un elemento esencial para el cultivo de café, sin embargo la mayoría de los suelos en Colombia presentan niveles bajos de este nutriente. La presencia de microorganismos solubilizadores de fosfatos (MSF) es una de las estrategias para suplir su demanda, en ese sentido se aislaron 26 bacterias rizosféricas de Typic melanudans de Cajibío (Cauca, Colombia), en tres agroecosistemas: café sin sombra, café con sombra y relicto de bosque secundario, evaluándoles la eficiencia solubilizadora de P (ESF) en Ca-P, Al-P, Fe-P en medios Pikovskaya sólido y líquido, obteniendo la secuencia de solubilización Ca-P > Al-P > Fe-P. Los dos aislamientos bacterianos con mayor ESF se identificaron por extracción del ADN y análisis del gen 16S rRNA como Kocuria sp, y Bacillus subtilis. Posteriormente se cuantificaron e identificaron los ácidos orgánicos presentes en las tres fuentes de fosforo por HPLC, siendo ellos los ácidos cítrico, glucónico, D- y L-málico, D- y L-láctico con mayor presencia en Ca-P- Kocuria sp. En todas las condiciones, se observó que ocurrió acidificación de los medios, siendo más fuerte en Fe-P y Al-P.

Abstract Phosphorus is an essential nutrient for coffee growing, however, in Colombia most of the soils have low concentrations of this element. A strategy to supply the demand is the use of phosphate solubilizing microorganisms (PSM), in that order, 26 rhizospheric bacteria of Typic melanudans soil of Cajibío (Cauca, Colombia) were isolated, in three agroecosystems: coffee at full sun exposure, coffee with shade and relic secondary forest, evaluating the phosphate solubilizing efficiency (PSE) for Ca-P, Al-P, Fe-P in the solid and liquid Pikovskaya media. It showed the following solubilization sequence: Ca-P> Al-P> Fe-P. Two isolated bacteria were identified by DNA extraction and analysis of the 16S rRNA gene as Kocuria sp, and Bacillus subtilis. Later, using HPLC were identified organic acids present in the three phosphorus sources as: citric, gluconic, D- and L-malic, D- and L-lactic acid with higher presence in Ca-P-Kocuria sp. Strong acidification was observed for Fe-P and Al-P in all conditions.

Evaluación de factores que afectan la bioacidulación de roca fosfórica bajo condiciones in vitro / Evaluation of factors that affect bioacidulation rock phosphate under in vitro conditions

Resumen El fósforo (P) es un nutriente esencial para el desarrollo de las plantas, desafortunadamente, su disponibilidad en muchos suelos es baja. Consecuentemente, los agricultores aplican altas cantidades de fertilizantes fosfóricos solubles, pero esto es ineficiente y costoso. El uso directo de roca fosfórica (RP) es muy atractivo por su bajo costo; sin embargo, es poco soluble y de baja eficiencia agronómica. Para superar esta limitación, hay un creciente interés en el uso de microorganismos del suelo capaces de disolverla y mejorar su valor como fertilizante. El objetivo de este trabajo fue evaluar el efecto que tienen algunos factores sobre la capacidad del hongo Mortierella sp. para disolver RP bajo condiciones in vitro. Estos factores son: (i) tiempo de incubación, (ii) tipo de RP, (iii) concentración inicial de P soluble y (iv) adición de vitaminas y micronutrientes. Despues del periodo de incubación se midió P en solución y pH. Los resultados indican que producto de la biodisolución de RP la más alta concentración de P en solución se alcanzó al día 5. Por otro lado, la biodisolución de RP fue reducida por la adición de vitaminas y micronutrientes y por el incremento en la concentración inicial de P soluble en el medio. Aunque la disolución microbiana fue más efectiva con la RP de Carolina del Norte, las RP del Huila y Santander presentaron un buen nivel de disolución en un periodo de tiempo corto. La bioacidulación mejorara la efectividad agronómica de la RP para su uso directo o a través de un proceso biotecnológico previo.

Abstract Phosphorus (P) is an essential nutrient for plant development, unfortunately, its availability in many soils is low. Consequently, farmers apply high quantities of soluble P fertilizers, but this is an inefficient and costly practice. The direct use of rock phosphate (RP) is a highly attractive option because its low cost, but this material has low solubility and low agronomic efficiency. In order to overcome this limitation, there is a growing interest in the use of soil microorganisms capable of dissolving RP and improving its value as a P fertilizer. The objective of this study was to evaluate the effect of some factors on the effectiveness of the fungus Mortierella sp. to dissolve RP under in vitro conditions. These factors included: (i) incubation time, (ii) type of RP, (iii) initial concentration of soluble P, and (iv) addition of vitamins and micronutrients. After the incubation period, P and pH were measured in solution. The results indicated that as a consequence of the biodissolution of RP, the highest concentration of soluble P in the medium was reached on the day 5th. The biodissolution of RP was reduced by the addition of vitamins and micronutrients and by the increase in the initial concentration of soluble P. Although microbial dissolution was more effective with North Carolina RP, RPs from Huila and Santander showed a good level of dissolution in a short period of time. Bioacidulation will improve the agronomic effectiveness of RP for its direct use or through a previous biotechnological process.

Assessment of Rice Associated Bacterial Ability to Enhance Rice Seed Germination and Rice Growth Promotion

ABSTRACT The application of beneficial bacteria has recently been used for sustainable agriculture. In current research, 71 bacterial isolates were obtained from rice plant and the rhizosphere soil of different paddy fields in Guilan province, Iran. After primitive investigation, 40 bacteria with typical predominant characteristics were selected. By PCR-RFLP of their 16S r-DNA gene, 8 Operational Taxonomic Units (OTUs) totally consisted of 33 isolates were obtained. From all of them, 8 isolates were selected for rice seed germination experiment, then, effective isolates were used for pot experiment to evaluate their ability for promoting rice growth. All of them were able to increase rice growth and yield, but in different potential. These tested isolates were identified as Alcaligenes faecalis (DEp8, O1R4), Pantoea ananatis (AEn1), Bacillus vietnamensis (MR5), Bacillus idriensis (MR2) and Stenotrophomonas maltophilia by partial sequencing of their 16S r-DNA gene. Among them, AEn1 and MR5 produced indole-3- acetic acid (IAA) in larger amounts than the other isolates and the isolates AEn1 and O1R4 were able to solubilize phosphate in higher amounts. According to the results obtained, it can be concluded that AEn1, O1R4 and MR5 can be considered as bacterial inoculants to use as alternatives for chemical fertilizers.

Agronomic efficiency and profitability of cotton on integrated use of phosphorus and plant microbes

Abstract Cotton crop, plays a significant role in Pakistans economy by ruling a prominent place in edible oil and local textile industry. Phosphorus (P) inaccessibility and deficiency of soil organic matter are the key restraints for low crop productivity in cotton. Therefore, a two years field study was designed during 2014-15, to explore the influence of phosphate solubilizing bacteria (PSB), farmyard manure (FYM), poultry manure (PM) and inanimate sources of P on various physiological, growth, yield and quality parameters of cotton crop at CCRI Multan. Field responses of seeds inoculated with two distinctive phosphate solubilizing bacteria (PSB) strains viz. S0 = control, S1 =strain-1, S2 = strain-2 and eight organic, inorganic P sources viz., P0= control, P1 = 80 kg ha-1 P from inorganic source, P2 = 80 kg ha-1 P from FYM, P3 = 80 kg ha-1 P from PM, P4 = 40 kg ha-1 P from FYM + 40 kg ha-1 P from inorganic source, P5 = 40 kg ha-1 P from PM + 40 kg ha-1 P from inorganic source, P6 = 80 kg ha-1 P from FYM + 40 kg ha-1 P from inorganic source, P7 = 80 kg ha-1 P from PM + 40 kg ha-1 P from inorganic source and P8 = 40 kg ha-1 P from FYM + 40 kg ha-1 P from PM were evaluated. Results revealed that inoculation of seeds with PSB and collective use of inorganic and organic sources of P had considerably increased the yield contributing attributes in cotton. However, the treatment P7 (80 kg P ha-1 from PM + 40 kg P ha-1 from inorganic source) in coincidence with seeds inoculated with PSB (S1) produced taller plant, maximum boll weight, significantly higher LAI and CGR. Significantly higher seed cotton yield, lint yield, fiber length and maximum BCR of 1.95 and 1.81 was also obtained from the P7 treatment during both crop-growing seasons. In conclusion, combined use of 80 kg P ha-1 from PM + 40 kg P ha-1 from inorganic source and cotton seeds inoculated with strain-1 improved phosphorus uptake ensuing in greater consumption of photo-assimilates for maximum growth and yield.

Resumo A safra de algodão, desempenha um papel significativo na economia do Paquistão, ao ocupar um lugar de destaque no óleo comestível e na indústria têxtil local. A inacessibilidade e a deficiência de fósforo (P) da matéria orgânica do solo são as principais restrições para a baixa produtividade das culturas em algodão. Portanto, um estudo de campo de dois anos foi desenvolvido durante 2014-15, para explorar a influência de bactérias solubilizantes de fosfato (PSB), esterco de capoeira (FYM), esterco de aves (PM) e fontes inanimadas de P sobre vários fatores fisiológicos, crescimento, rendimento e parâmetros de qualidade da cultura do algodão no CCRI Multan. Respostas de campo de sementes inoculadas com duas cepas distintas de bactérias solubilizantes de fosfato (PSB) viz. S0 = controle, S1 = cepa-1, S2 = cepa-2 e oito fontes orgânicas de P inorgânicas viz., P0 = controle, P1 = 80 kg ha-1 P de fonte inorgânica, P2 = 80 kg ha-1 P de FYM, P3 = 80 kg ha-1 P de PM, P4 = 40 kg ha-1 P de FYM + 40 kg ha-1 P de fonte inorgânica, P5 = 40 kg ha-1 P de PM + 40 kg ha-1 P de fonte inorgânica, P6 = 80 kg ha-1 P de FYM + 40 kg ha-1 P de fonte inorgânica, P7 = 80 kg ha-1 P de PM + 40 kg ha-1 P de fonte inorgânica e P8 = 40 Foram avaliados kg ha-1 P da FYM + 40 kg ha-1 P da PM. Os resultados revelaram que a inoculação de sementes com PSB e o uso coletivo de fontes inorgânicas e orgânicas de P aumentaram consideravelmente os atributos que contribuem para a produtividade no algodão. No entanto, o tratamento P7 (80 kg P ha-1 da PM + 40 kg P ha-1 da fonte inorgânica) em coincidência com sementes inoculadas com PSB (S1) produziu planta mais alta, peso máximo de cápsula, IAF e CGR significativamente maiores. Produtividade significativamente maior do algodão, rendimento de fiapos, comprimento da fibra e BCR máximo de 1,95 e 1,81 também foi obtida a partir do tratamento P7 durante as duas épocas de cultivo. Em conclusão, o uso combinado de 80 kg P ha-1 de PM + 40 kg P ha-1 de fonte inorgânica e sementes de algodão inoculadas com a cepa-1 melhoraram a captação de fósforo, resultando em maior consumo de foto assimilados para obter crescimento e produtividade máximos.

Characterization of phosphate solubilizing bacteria and nitrogen fixing bacteria from limestone mining region

Aims: Phosphate and nitrogen are major macronutrients needed by plants. Phosphates in the soil are present in the organic and inorganic form. The amounts of phosphate and nitrogen in marginal soil can be increased by plant growth promoting rhizobacteria (PGPR). The aim of this study was to isolate and characterize phosphate solubilizing bacteria which has ability to fix nitrogen from the soil around limestone mining area. Methodology and results: There were 22 isolates that could solubilize phosphate and fix nitrogen. There were 9 isolates that could not cause hypersensitivity symptoms (necrotic) on tobacco leaf. Quantitative assay of phosphate solubilizing was done by colorimetric method. Quantitative assay of phosphate showed that isolate GPC1.7 had the highest phosphate solubilizing activity on Pikovskaya broth (450 mg/L) on the 6th and 7th day of incubation whereas isolate GPA2.2 had the highest nitrogen fixing activity (0.162 ppm/h), measured with Acetylene Reduction Assay whereas nitrogenase activity of GPC1.7 was unidentified. Isolate GPA2.1 and GPA2.2 were Gram negative bacteria whereas isolate GPC1.7 was Gram positive bacteria. Identification based on 16S rRNA gene showed that GPA2.1 was closely related to Pseudomonas psychrotolerans, GPA2.2 was closely related to Stenotrophomonas maltophilia, GPC1.7 was closely related to Bacillus megaterium and B. aryabhattai. Conclusion, significance and impact of study: Phosphate solubilizing bacteria and nitrogen fixing bacteria isolate could be further used for revegetation process of the ex-limestone mining area.

Caracterización de rizobacterias promotoras de crecimiento en plántulas de Eucalyptus nitens / Characterization of growth-promoting rhizobacteria in Eucalyptus nitens seedlings

Se aislaron bacterias rizosféricas y endófitas a partir de rizósfera y tejidos de raíz de árboles de Eucalyptus nitens con el objetivo de evaluar su capacidad de promover el crecimiento en plántulas de la misma especie en condiciones de invernadero. Los aislamientos que incrementaron el crecimiento de las plántulas fueron identificados y caracterizados por su capacidad de producir ácido indolacético (AIA), solubilizar fosfato y expresar la 1-aminociclopropano-1-carboxilato (ACC) desaminasa. Los 105 aislamientos obtenidos fueron morfológicamente diferentes y solo 15 promovieron significativamente el crecimiento de plántulas de E. nitens. Los máximos incrementos observados fueron en el peso seco aéreo (142 %) y de la raíz (135 %); también aumentaron la altura de las plantas (50 %) y el largo de raíces (45 %) de las mismas. Las rizobacterias pertenecieron a los géneros Arthrobacter, Lysinibacillus, Rahnella y Bacillus. Los aislados identificados como A. phenanthrenivorans 21 y B. cereus 113 incrementaron la emergencia de E. nitens a los 12 días en un valor promedio de 3,15 veces con relación al control. R. aquatilis aislado 78 presentó la mayor producción de AIA (97,5 ± 2,87 μg/ml) en presencia de triptófano y el mayor índice de solubilización de fósforo (2,4). B. amyloliquefaciens aislado 60 fue positivo para la actividad ACC desaminasa. Los resultados obtenidos indican el potencial de las rizobacterias estudiadas como promotoras de emergencia y crecimiento de plántulas de E. nitens y su posible uso como inoculantes, ya que presentan más de un mecanismo de acción asociado a la promoción del crecimiento.

Rhizospheric and endophytic bacteria were isolated from the rizosphere and root tissue of Eucalyptus nitens. The objective of this work was to evaluate their capacity to promote growth in seedlings of the same species under greenhouse conditions. The isolates that improved seedling growth were identified and characterized by their capacity to produce indoleacetic acid (IAA), solubilize phosphates and increase 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. One hundred and five morphologically different strains were isolated, 15 of which promoted E. nitens seedling growth, significantly increasing the height (50%), root length (45%) as well as the aerial and root dry weight (142% and 135% respectively) of the plants. Bacteria belonged to the genus Arthrobacter, Lysinibacillus, Rahnella and Bacillus. Isolates A. phenanthrenivorans 21 and B. cereus 113 improved 3.15 times the emergence of E. nitens after 12 days, compared to control samples. Among isolated R. aquatilis, 78 showed the highest production of IAA (97.5±2.87 μg/ml) in the presence of tryptophan and the highest solubilizer index (2.4) for phosphorus, while B. amyloliquefaciens 60 isolate was positive for ACC deaminase activity. Our results reveal the potential of the studied rhizobacteria as promoters of emergence and seedling growth of E. nitens, and their possible use as PGPR inoculants, since they have more than one mechanism associated with plant growth promotion.

A study on nitrogen fixing and phosphate solubilizing microorganisms on growth and Physiology of Plumbago Zeylanica.L.

Organisms that are commonly used as biofertilizers component are nitrogen fixers (N-fixer), potassium solubilizer (K-solubilizer) and phosphorus solubilizer (P- solubilizer), or with the combination of molds or fungi. Most of the bacteria included in biofertilizer have close relationship with plant roots. In this work we have selected plumbago zeylanica.L plant to study the effect of Azotobacter on the growth of roots, stem, and leaves. Also biochemical characterization was done to identify the effect of Azotobacter in Plumbago. The maximum shoot length was recorded in T4 plants (43.51) on 90th days of plant growth after transplanting the plants. There was a significant increase at 5 % level in the root length from 30th days to 90th days in all the treatments. The maximum number of leaves were found in T4 treatment followed by T3 and T2. Minimum numbers of leaves were found in T1 (1083). On 60th day and 90th day also the total chlorophyll content was maximum in T4 treated plants followed by T3, T2 plants. The amount of reducing sugars (μg/g) in shoots of T4, T3 and T2 plants on 30th, 60th and 90th days were significantly high when compared to T1 plants. The content of protein in roots of T2, T3 and T4 plants on 30th, 60th and 90th days were significantly high when compared to protein content of T1 plants.

Diseño de un medio para la producción de un co־cultivo de bacterias fosfato solubilizadoras con actividad fosfatasa / Design of a culture media for the production of a phosphate-solubilizing bacteria co-culture / Desenho de um meio para a produção de uma co-cultura de bactérias solubilizadoras de fosfato com actividade da fosfatase

Objetivo. Diseñar un medio de cultivo complejo para la producción de biomasa y fosfatasas ácidas a partir de bacterias solubilizadoras de fosfatos aisladas de suelo. Materiales y métodos. A partir de muestras de suelo de cultivos de palma de aceite se realizaron los aislamientos y la selección de bacterias fosfato solubilizadoras (BFS) en agar SMRS1, las cuales fueron sometidas a pruebas de antagonismo con el fin de verificar su aptitud para la formación de co-cultivos. Posteriormente, se realizó un diseño experimental Box-Behnken para evaluar el efecto de cada uno de los componentes del medio de cultivo sobre la producción de biomasa y enzimas fosfatasas a escala de laboratorio. Finalmente se realizaron curvas de crecimiento y de producción de enzima para determinar los tiempos de producción. Resultados. Se obtuvieron 5 bacterias fosfato solubilizadoras, de las cuales 3 fueron seleccionadas con base en el índice de solubilización; dichas cepas, de morfología bacilar Gram negativa, fueron identificadas como A, B y C, cuyos índices de solubilización correspondieron a 2,03, 2,12 y 2,83, respectivamente. De acuerdo con los análisis de ANOVA para el diseño experimental de Box Behnken, el factor que tuvo efecto significativo sobre la actividad fosfatasa (p<0,01), fue el hidrolizado de levadura, y el formulado que generó la mayor concentración de biomasa y actividad fosfatasa (p<0,01) fue el que contenía 10, 15 y 2,5 gL-1 de roca fosfórica sacarosa e hidrolizado de levadura, respectivamente, obteniendo valores máximos de biomasa y actividad fosfatasa de 11,8 unidades logarítmicas de UFC y 12,9 unidades fosfatasa con incubación por 24 horas a 100 rpm. Conclusión. Se determinó que el medio con formulación 10gL-1 de roca fosfórica, 2,5gL-1 de hidrolizado de levadura y 15gL-1 de sacarosa comercial, fue ideal para la producción de biomasa y enzimas fosfatasas a partir de las cepas evaluadas. Así mismo, se comprobó que el hidrolizado de levadura fue el único factor significativamente influyente en la producción de enzimas fosfatasas.

Objective. To design a complex culture media for the production of biomass and acid phosphatases from phosphate-solubilizing bacteria isolated from soil. Materials and methods. Phosphate-solubilizing bacteria were isolated from oil palm crop soil samples and selected on SMRS1 agar, which were then assessed with antagonism tests to verify their aptitude to form a co-culture. A Box-Behnken experimental design was applied to evaluate the effect of each one of the culture media components on the production of biomass and phosphatase enzymes at a laboratory scale. Finally, microbial growth and enzyme production curves were carried out in order to determine their production times. Results. Five phosphate-solubilizing bacterial strains were isolated and three of them were selected based on their solubilization indices. These Gram negative strains with bacillus morphology were identified as A, B and C; their solubilization indices were 2.03, 2.12, and 2.83, respectively. According to the ANOVA analyses for the Box-Behnken design, the only factor which had a significant effect on the phosphatase activity (p<0.01) was hydrolyzed yeast, and the formulation that generated the highest biomass concentration and phosphatase activity (p<0.01) contained 10, 15 and 2.5 gL-1 of phosphoric rock, sucrose and hydrolyzed yeast, respectively. After 24 hours of incubation at 100 rpm, the highest values of biomass and phosphatase activity were obtained: 11.8 logarithmic units of CFU and 12.9 phosphatase units. Conclusion. We determined that the culture media based on phosphoric rock 10 gL-1, hydrolyzed yeast 2.5 gL-1 and commercial sucrose 15 gL-1 was ideal for the production of biomass and phosphatases by the strains evaluated; likewise, we proved that the hydrolyzed yeast was the only factor significantly influential for the production of phosphatases.

Objetivo. Desenhar um meio de cultura complexo para a produção de biomassa e fosfatase ácida a partir de bactérias solubilizadoras de fosfato isoladas do solo. Materiais e métodos. De amostras de solo de plantações de dendezeiros foram isoladas e selecionadas bactérias solubilizadoras de fosfato (BFS) em ágar SMRS1, que foram testadas em provas de antagonismo para verificar sua capacidade de formar co-culturas. Subsequentemente, foi realizado um desenho experimental do tipo Box-Behnken para avaliar o efeito de cada um dos componentes do meio de cultura na produção de biomassa e de enzimas fosfatase a escala de laboratório. Finalmente foram realizadas curvas de crescimento e de produção da enzima para determinar os tempos de produção. Resultados. Foram obtidas 5 bactérias solubilizadoras de fosfato, das quais 3 foram selecionadas com base no índice de solubilização, tais cepas, de morfologia bacilar Gram negativas, foram identificadas como A, B e C, cujos índices de solubilização corresponderam a 2,03, 2 12 e 2,83, respectivamente. De acordo com a análise ANOVA para o desenho experimental do tipo Box Behnken, o fator que teve efeito significativo na atividade da fosfatase (p <0,01), foi o hidrolisado de levedura, e o formulado que gerou a maior concentração de biomassa e atividade da fosfatase (p <0,01) foi aquel que contive 10, 15 e 2,5 gL-1 de rocha fosfato sacarose e hidrolisado de levedura, respectivamente, obtendo-se valores máximos de biomassa e atividade de fosfatase de 11,8 unidades log de UFC e 12,9 unidades de fosfatase com incubação durante 24 horas a 100 rpm. Conclusão. Foi determinado que o meio com formulação 10gL-1 de rocha fosfórica, 2,5 gL-1 de hidrolisado de levedura e 15gL-1 de sacarose comercial, foi ideal para a produção de biomassa e enzimas fosfatase a partir das cepas avaliadas. Da mesma forma, verificou-se que o hidrolisado de levedura foi o único fator significativo influente na produção de enzimas fosfatase.

Role of phosphate solubilizing bacteria on rock phosphate solubility and growth of aerobic rice.

Use of phosphate-solubilizing bacteria (PSB) as inoculants has concurrently increased phosphorous uptake in plants and improved yields in several crop species. The ability of PSB to improve growth of aerobic rice (Oryza sativa L.) through enhanced phosphorus (P) uptake from Christmas island rock phosphate (RP) was studied in glasshouse experiments. Two isolated PSB strains; Bacillus spp. PSB9 and PSB16, were evaluated with RP treatments at 0, 30 and 60 kg ha-1. Surface sterilized seeds of aerobic rice were planted in plastic pots containing 3 kg soil and the effect of treatments incorporated at planting were observed over 60 days of growth. The isolated PSB strains (PSB9 and PSB16) solubilized significantly high amounts of P (20.05-24.08 mg kg-1) compared to non-inoculated (19-23.10 mg kg-1) treatments. Significantly higher P solubilization (24.08 mg kg-1) and plant P uptake (5.31 mg plant -1) was observed with the PSB16 strain at the highest P level of 60 kg ha-1. The higher amounts of soluble P in the soil solution increased P uptake in plants and resulted in higher plant biomass (21.48 g plant -1). PSB strains also increased plant height (80 cm) and improved root morphology in aerobic rice. The results showed that inoculation of aerobic rice with PSB improved phosphate solubilizing activity of incorporated RP.

Primary Identification, Capability of Phosphate-Solubilization and Optimization of Medium of Some Microorganism from Mangrove / 微生物学通报

The genetic indentification of 16S rDNA or ITS, capability of phosphate-solubilization and pH of medium, and optimization of medium of some microorganism isolated from mangrove were investigated in this study. The result showed that the fungi normally had much higher capacity to dissolve the inorganic phosphate than the bacteria, the capacity of the fungi was closely correlated to the pH of medium, but the relationship was weak for the bacteria. It was illustrated by single factor experiments that the optimal carbon and nitrogen sources were maltose and urea respectively. The orthogonal design was employed in testing the optimum composition of medium composed of 5 g/L maltose, 0.05 g/L urea, 5 g/L NaCl, pH 5. In this optimal medium, the effectively enrichment of bacteria could reach up to 6.06?109 CFU/mL under 30?C for 48 hours cultivation.