Combined effect of Pseudomonas sp. and Trichoderma aureoviride on lettuce growth promotion

Plant growth promotion by microorganisms may be a viable alternative to increase lettuce production through pathogens control and nutrients absorption increase. Trichoderma and Pseudomonas genus are examples of widely studied microorganisms with the capacity to promote plant growth. However, there are still gaps regarding the action of the combined effect of these two microorganisms. Therefore, the objective of this study was to evaluate the combined effect of Pseudomonas sp. UAGF14 and Trichoderma aureoviride URM5158 on the development of lettuce plants. The experimental design was completely randomized, with five treatments: CONT (control), CM (soil with organic fertilization), CMB (soil with organic fertilization and Pseudomonas sp.), CMF (soil with organic fertilization and T. aureoviride), and CMFB (soil with organic fertilization, Pseudomonas sp. and T. aureoviride), with ten repetitions. At 30, 40 and 60 days after sowing, the following parameters were analyzed: plant and canopy height and number of leaves. At 60 days after emergence, shoot dry matter, leaf area, root dry matter, root length and chlorophyll were analyzed. Catalase, peroxidase and polyphenol oxidase enzymatic activity were determined. The CMFB treatment had the highest means of lettuce growth promotion, confirming the synergistic effect of the combination of the two microorganism types, as it increased height, canopy, shoot and root dry matter, and chlorophyll levels compared to CONT, although did not differ from CM in some variables. Enzymatic activity was also influenced by the action of these microorganisms combined, evidencing by polyphenol oxidase increase. The CMFB or CM were efficient in promoting lettuce growth, showing positive response to the plant morphological and physiological characteristics. However, few responses were observed in lettuce plant growth in the first cycle evaluated after 60 days, compared CM and CMFB treatments, but both treatments showed superiority in lettuce plant growth submitted to CONT treatment. Therefore, further studies are needed to estimate the long-term effects of combined effect of Pseudomonas sp. UAGF14 and T. aureoviride URM5158 on crop productivity in field conditions

Diazotrophic Bacterial Community of Degraded Pastures

Pasture degradation can cause changes in diazotrophic bacterial communities. Thus, this study aimed to evaluate the culturable and total diazotrophic bacterial community, associated with regions of the rhizosphere and roots of Brachiaria decumbens Stapf. pastures in different stages of degradation. Samples of roots and rhizospheric soil were collected from slightly, partially, and highly degraded pastures. McCrady’s table was used to obtain the Most Probable Number (MPN) of bacteria per gram of sample, in order to determine population density and calculate the Shannon-Weaver diversity index. The diversity of total diazotrophic bacterial community was determined by the technique of Denaturing Gradient Gel Electrophoresis (DGGE) of the nifH gene, while the diversity of the culturable diazotrophic bacteria was determined by the Polymerase Chain Reaction (BOX-PCR) technique. The increase in the degradation stage of the B. decumbens Stapf. pasture did not reduce the population density of the cultivated diazotrophic bacterial community, suggesting that the degradation at any degree of severity was highly harmful to the bacteria. The structure of the total diazotrophic bacterial community associated with B. decumbens Stapf. was altered by the pasture degradation stage, suggesting a high adaptive capacity of the bacteria to altered environments

Genetic variability in regenerated Metarhizium flavoviride protoplasts

Protoplast isolation and regeneration were evaluated in two wild-type and two colour mutant strains of Metarhizium flavoviride. Cultivation in liquid medium, followed by mycelium treatment with Novozym 234 in the presence of KCl 0.7M as osmotic stabilizer, produced 5.05 x 10(6) to 1.15 x 10(7)x mL-1 protoplasts. The percentage of regeneration ranged from 6.65 to 27.92%. Following protoplast regeneration, one strain produced spontaneously stable morphological variant colonies. Although colonies with altered morphology have been reported in bacteria following protoplast regeneration, this is the first time that the same is described in a filamentous fungus. The original strain and one derived variant were tested for sensitivity to the fungicides benomyl and captan.<br>A formação e regeneração de protoplastos foram avaliadas em duas linhagens selvagens e duas linhagens mutantes para coloração de conídios em Metarhizium flavoviride. O cultivo em meio líquido seguido do tratamento do micélio com Novozym 234 na presença de KCl 0,7 M como estabilizador osmótico, resultou na produção de 5,05´10(6) a 1,15´10(7) protoplastos´mL-1. A porcentagem de regeneração das diferentes linhagens variou de 6,65 a 27,92%. Após a regeneração, uma das linhagens selvagens produziu espontaneamente variantes estáveis, com morfologia alterada. Embora variantes morfológicos já tenham sido observados após regeneração de protoplastos em bactérias, esta parece ser a primeira vez que tal ocorrência é descrita em fungos filamentosos. Um desses variantes, além da linhagem selvagem da qual ele foi originado, foi testado para sensibilidade aos fungicidas benomil e captano

EXOPOLYSACCHARIDES AND ABIOTIC STRESS TOLERANCE IN BACTERIAL ISOLATES FROM “SABIÁ” NODULES

Several microorganisms produce polysaccharides, deemed to protect the bacteria from several environmental stresses. This paper aims to evaluate the protective effect of exopolyssacharides to different abi- otic stresses in bacterial isolates from “sabiá” ( Mimosa caesalpiniifolia ) nodules. 303 fast growing isolates were qualitatively evaluated for exopolyssacharide production and tested in vitro for tolerance to two levels of acidity, joint aluminum and acidity, three salinity levels; 11 antibiotics and three herbicides. Most isolates re- sisted media acidity, acidity with aluminum, salinity, and ampicillin, cefotaxime, gentamicin and vancomycin antibiotics and 2,4D herbicide, while being sensitive to ciprofloxacin, chloramphenicol, streptomycin, kanamy- cin, nalidixic acid, rifampicin and tetracycline antibiotics and paraquat and glyphosate herbicides. There was no connection between exopolyssacharide production and abiotic stress tolerance

Evaluation methods used for phosphate-solubilizing bacteria

Phosphorus solubilizers naturally acidify rhizospheric soil and increase phosphorus availability; therefore, their evaluation may help to reduce phosphorus fertilizer use. This work aimed to evaluate the different selection methods and select inorganic phosphorus-solubilizing bacteria as potential plant-growth promoters. Bacterial isolates obtained from sugarcane roots and soil were tested using solid growth media containing bicalcium phosphate and Irecê Apatite ground rock phosphate as phosphorus sources. Seven isolates with high (3), moderate (3) and low solubilization indices (1) and the Pseudomonas fluorescens R-243 strain were tested in two liquid growth media, followed by the pH and soluble P in the solution. The same isolates, in the absence of inoculation, were tested in Leonard jars with two high- and low-solubility sources using cowpea as a test species. Forty-four days after planting aboveground dry mass, the phosphorus content and total aboveground phosphorus and substratum phosphorus contents were evaluated. The growth media affected phosphorus solubilization by the bacteria. Evaluation of liquid media was the most reliable method for analyzing bicalcic phosphorus solubilization by the bacteria not linked to pH reduction. Isolates UAGC 17, 19 and 65 should be better studied because they were the best solubilizers in culture media; however, they did not demonstrate the same efficiency when inoculated on cowpea.Keywords: Solubilization, phosphate, P-solubilizing-microorganism

Isolamento e prospecção de bactérias endofíticas e epifíticas na cana-de-açúcar em áreas com e sem cupinicida

A exploração e o conhecimento dos mecanismos de interação bactéria-planta podem ser utilizados para a produção sustentável de diversas culturas, como a cana-de-açúcar. O objetivo deste trabalho foi isolar e prospectar bactérias associadas à cultura da cana-de-açúcar com capacidade de fixar nitrogênio e solubilizar fosfato inorgânico em áreas com e sem aplicação de cupinicida. Bactérias endofíticas, de folha e raiz, e do rizoplano foram isoladas de três variedades comerciais (RB 92579, RB 867515 e RB 863129) de cana-de-açúcar, cultivadas no Nordeste brasileiro, em áreas com e sem aplicação de cupinicida, aos quatro meses após o plantio. Foram obtidos 272 isolados, selecionados quanto à sua capacidade de solubilizar fosfato inorgânico e fixar nitrogênio. O uso do cupinicida não provocou alterações nas comunidades microbianas endofíticas de folha e raiz e epifíticas do rizoplano. As variedades estudadas apresentam uma comunidade bacteriana associada com potencial aplicação para a promoção de crescimento vegetal, pois possuem a capacidade de fixar N2 e solubilizar fosfato inorgânico

Isolation, bioprospecting and diversity of salt-tolerant bacteria associated with sugarcane in soils of Pernambuco, Brazil

ABSTRACT The selection of salinity tolerant bacteria may provide greater promotion of plant growth in soils with salt stress. This study aimed to isolate and select bacterial isolates of sugarcane, tolerant to salinity and plant growth promoters, and assess genetic diversity. To do so, endophytic root and rhizosphere of sugarcane, were isolated in culture medium with and without NaCl. The bacterial population density, the potential for biological nitrogen fixation (BNF), production of indole acetic acid (IAA), inorganic phosphate solubilization and quorum sensing molecule production were evaluated. Furthermore, the genetic diversity was assessed by Denaturing Gradient Gel Electrophoresis (DGGE) of nifH. Salinity tolerance tests were also conducted. There was no statistical difference in population density. As for the potential for BNF, IAA production, phosphate solubilization and quorum sensing molecule production, there was a high percentage of positive isolates. In the technique of DGGE nifH, the dendrogram showed high genetic variability. So there was the selection of bacteria that can contribute to the better development of sugarcane in saline soils

Bioprospection and genetic diversity of endophytic bacteria associated with cassava plant

<div><p>ABSTRACT Cassava is mostly planted in sandy soils which are usually of low fertility, thereby making it necessary to perform beneficial associations with microorganisms that can promote their growth. In this perspective, the possibility of selecting bacterial isolates efficient in promoting the growth of the culture is evident, which can provide subsidies for future inoculants. The objective of this study was to isolate, identify, select and evaluate the genetic diversity of endophytic bacteria in roots and stems of cassava grown in Garanhuns - PE, with features involved in promoting plant growth. The isolation was performed on culture medium semisolid LGI-P. The selected isolates were evaluated for the potential to fix N2, as the ability to produce indole acetic acid, for their ability to solubilize inorganic phosphate and produce exopolysaccharides. Some bacterial isolates had their 16S rRNA gene sequenced by the Sanger method. A total of 52 endophytic bacteria isolates were obtained from cassava. Regarding the potential to fix N2, 15% of the isolates were positive. As for the production of IAA, 78% of the isolates produced this phytohormone in a medium with increased L-tryptophan. Approximately 31% of the isolates were able to solubilize inorganic phosphate and 60% had exopolysaccharide. The identification of 19 isolates allowed the grouping into six bacterial genera, namely: Achromobacter, Bacillus, Burkholderia, Enterobacter, Pantoea and Pseudomonas. Cassava plants grown in Garanhuns - PE present interaction with different groups of endophytic bacteria and there are bacterial groups with several characteristics involved in promoting plant growth.</p></div

Diazotrophic bacteria isolated from Brachiaria spp.: genetic and physiological diversity

Resumen Pastos del género Brachiaria spp. predomina en pasturas con suelos poco fértiles. Este escenario resalta la importancia de la asociación con microorganismos para fomentar el cultivo de plantas. Este estudio tuvo como objetivo evaluar la variabilidad genética e identificar los mecanismos de promoción del crecimiento de las plantas, in vitro, de bacterias asociadas con Brachiaria decumbens Stapf. y Brachiaria humidicola (Rendle.) Schweickerdt en Pernambuco, Brasil. Evaluamos 20 aislamientos de bacterias diazotróficas obtenidas de la comunidad de endofitas o rizosfera. Las características genéticas se determinaron mediante la secuenciación de la región 16S rRNA, lo que permitió identificar diez géneros bacterianos diferentes: Bacillus sp., Burkholderia sp., Enterobacter sp., Klebsiella sp., Microbacterium sp., Pantoea sp., Ralstonia sp., Rhizobium sp., Sinomonas sp., y Sphingomonas sp., con una especificidad del género Rhizobium sp. a Brachiaria decumbens Stapf.. Las características fenotípicas y funcionales revelaron que el 100% de las cepas bacterianas producían ácido indol-3-acético (AIA) con la adición de L-triptófano y el 60% presentaban producción de AIA independientemente de la vía de L-triptófano. También detectamos que el 70% de las bacterias aisladas poseían la capacidad de solubilizar el fósforo. El análisis de la producción enzimática reveló que el 30% de los aislados producían celulasa, 60% pectato liasa, 15% poligalacturonasa y el 30% producía amilasa. También detectamos la producción de N-acyl homoserine lactones en el 65% de las cepas bacterianas. En resumen, las plantas de B. decumbens Stapf. y B. humidicola (Rendle.) Schweickerdt interactuó con diferentes géneros de bacterias capaces de promover el crecimiento de la planta.Abstract Grass from the genus Brachiaria spp. predominates in pastures with low fertile soils. This scenario highlights the importance of the association with microorganisms to foster plant growth, which becomes essential to the successful establishment of this forage in such environments. This study aimed to evaluate the genetic variability and identify the mechanisms of plant growth promotion, in vitro, of bacteria associated with Brachiaria decumbens Stapf. and Brachiaria humidicola (Rendle.) Schweickerdt in Pernambuco, Brazil. We evaluated 20 isolates of diazotrophic bacteria obtained from the endophyte or rhizosphere communities. The genetic characteristics were determined via sequencing the 16S rRNA region, which allowed us to identify ten different bacterial genera: Bacillus sp., Burkholderia sp., Enterobacter sp., Klebsiella sp., Microbacterium sp., Pantoea sp., Ralstonia sp., Rhizobium sp., Sinomonas sp., and Sphingomonas sp., with a specificity of the genus Rhizobium sp. to Brachiaria decumbens Stapf.. The phenotypic and functional characteristics revealed that 100% of the bacterial strains produced indol-3-acetic acid (IAA) with the addition of L-tryptophan, and 60% presented IAA production independent of the L-tryptophan pathway. We also detected that 70% of the isolated bacteria possessed the capacity to solubilize phosphorus. The analysis of the enzymatic output revealed that 30% of the bacterial isolates produced cellulase, 60% produced pectate lyase, 15% produced polygalacturonase, and 30% produced amylase. We also detected the production of N-acyl homoserine lactones in 65% of bacterial strains. In summary, our results showed that plants of B. decumbens Stapf. and B. humidicola (Rendle.) Schweickerdt interacted with different bacterial genera capable of promoting plant growth