Ensiling pretreatment fortified with laccase and microbial inoculants enhances biomass preservation and bioethanol production of alfalfa stems.

This study investigated the potential of ensiling pretreatment fortified with laccase and a lactic acid bacteria (LAB) inoculant on improving the utilization of alfalfa stems for bioethanol production. The alfalfa stems were ensiled with no additives (Con), 0.04 % laccase (LA), a LAB inoculant containing Pediococcus pentosaceus at 1 × 106 fresh weight (FW) and Pediococcus acidilactici at 3 × 105 cfu/g FW (PP), and a combination of LA and PP (LAP) for 120 days. By reshaping the bacterial community structure of alfalfa stem silages toward a higher abundance of Lactobacillus, the addition of laccase and LAB inoculant either alone or in combination facilitated lactic acid fermentation to reduce fermentation losses, as evidenced by low concentrations of ammonia nitrogen (53.7 to 68.9 g/kg total nitrogen) and ethanol (2.63 to 3.55 g/kg dry matter). All additive treatments increased lignocellulose degradation and soluble sugars concentrations of alfalfa stem silages. Due to delignification and polyphenol removal, glucan and xylan conversion (70.3 % vs. 35.7 % and 51.6 % vs. 27.9 %, respectively) and ethanol conversion efficiency (53.9 % vs. 26.4 %) of alfalfa stems were greatly increased by ensiling fortified with LA versus Con, and these variables (79.8 % for glucan, 58.7 % for xylan, and 60.1 % for ethanol conversion efficiency) were further enhanced with a synergistic effect of LA and PP fortification. The spearman correlation analysis revealed that bioethanol fermentation of silage biomass was closely related to ensiling parameters and total phenols. In conclusion, ensiling pretreatment with LA and PP combination offered a feasible way to efficient utilization of alfalfa stems for bioethanol production.

Novel sulphur-oxidizing bacteria consummate sulphur deficiency in oil seed crop.

Plants absorb sulphate, the oxidized form of elemental sulphur (S°), from soil. Sulphur-oxidizing bacteria play a key role in transformation of sulphur in soil. Oil seed crops require high amount of sulphur and it plays an important role in the formation of proteins, vitamins and enzymes. It increases yield, oil content and protein content in oil seed crops. Sulphur is the important constituent of amino acids, viz. methionine, cystine, and cysteine. It necessitates various enzymatic, metabolic processes such as photosynthesis and nitrogen fixation. In the last few years, the prominence of sulphur in oil seed crop nutrition has been accepted as widespread occurrence of its inadequacy in agricultural soil. Approximately 41% of Indian soil is deficient in sulphur. The soil microbial population is the major enforcement behind sulphur transformation. They mineralize, immobilize, oxidize and reduce the elemental and other reduced forms of sulphur. The main step in transformation is oxidation carried out by microorganisms to convert sulphur into sulphate. The chemolithotrophic bacteria belonging to genus Thiobacillus are of primary importance; there are heterotrophic bacteria also which can oxidize sulphur in soil. The pH reduction at the time of oxidation helps in mineralization and absorption of other essential nutrients also. This property of sulphur-oxidizing bacteria (SOB) shows their potential to be used as bioinoculants. Bioformulations prepared using carrier-based formulations, immobilization, biostimulation, etc., are sustainable forms of fertilizers. These SOB inoculants can be used to increase the fertility and sulphate production in soil.

Bioaugmentation: possible scenarios due to application of bacterial preparations for remediation of oil-contaminated soil.

Although bioaugmentation is known as effective and environmentally friendly method increasing removal of hydrocarbons from oil-contaminated soil, it sometimes fails in soil restoration and disturbs the ecological state of soil. We studied possible scenarios of the introduction of oil-degrading bacteria into oil-contaminated podzolic soil assessing the environmental safety of different bacterial preparations in a long-term field experiment. Integral indicators characterizing the state of biocenosis included biological activity of soil and aboveground biomass of grasses. It has been established that bacterial preparations can have both positive and negative effects on the ecological state of soil and oil biodegradation. Of the five bacterial preparations studied, one had a pronounced positive effect on soil biological activity and oil mineralization processes. Two preparations did not accelerate oil biodegradation and were characterized by a weaker positive effect or even a lack of influence. Two more bacterial preparations had a significant negative impact on soil biological properties. These preparations slowed oil mineralization in soil. Both positive and negative effects of bacterial preparations were observed only during the first two years after their application. All preparations were not effective during the latter stages of long-term remediation processes. The results indicate that successful application of bioaugmentation for the restoration of oil-contaminated soil requires testing of environmental safety of bacterial preparations in a long-term field experiments prior to any treatment processes.

Stage-dependent concomitant microbial fortification improves soil nutrient status, plant growth, antioxidative defense system and gene expression in rice.

Stage-dependent concomitant fortification of rice (Oryza sativa L.) varieties PB1612 and CO51 with microbial inoculants Trichoderma asperellum and Pseudomonas fluorescens as seed coating, seedling root inoculation and soil application enhanced growth, activated antioxidant enzymes and modulated defence-related genes in plants. Microbial inoculants improved shoot height, tiller numbers, fresh weight and dry biomass. Co-inoculation was more impactful in enhancing plant growth and development as compared to single inoculation. Single and co-inoculation improved organic carbon (OC) and N, P and K content in the soil substantially. Mean values between control and co-inoculation varied significantly for OC in PB1612 (p0.001) and CO51 (p0.019) and phosphorus content in PB1612 (p0.044) and CO51 (p0.021). Microbial inoculation enhanced soil nutrients and increased their bioavailability for the plants. Total polyphenolics, flavonoids and protein content increased in the leaves following microbial inoculation. Enhanced non-enzymatic antioxidant parameters (ABTS, DPPH, Fe-ion reducing power and Fe-ion chelation) was found in microbe inoculated rice reflecting high free radical scavenging activity in polyphenolics-rich leaf extracts. Increased enzyme activity of superoxide dismutase (SOD), glutathione reductase (GR), phenylalanine ammonia-lyase (PAL), peroxidase (PO), glutathione peroxidase (GPX), ascorbate peroxidase (APX) and catalase (CAT) showed improved ROS scavenging in rice plants having co-inoculation. Over-expression of PAL, cCuZn-SOD and CAT genes in microbial inoculated rice plants was recorded. The study concludes that plant stage-wise concomitant fortification by microbial inoculants could play multi-pronged manifestations at physiological, biochemical and molecular level in rice to positively influence growth, development and defense attributes in plants.

Inoculation of Cd-contaminated paddy soil with biochar-supported microbial cell composite: A novel approach to reducing cadmium accumulation in rice grains.

Bioremediation of heavy metal-contaminated soil using metal-resistant microbes is a promising remediation technology. However, as exogenous bacteria sometimes struggle to survive and grow when introduced to new soils, it is important to develop appropriate carriers for microbial populations. In this study, we report a novel approach to remediating Cd-contaminated rice paddy soil using biochar-supported microbial cell composites (BMCs) produced from agricultural waste (cornstalks). Pot experiments showed that amendment with BMC was more efficient at reducing root and grain Cd content than pure bacteria, while improving soil Cd fractionation toward more stabilized and less labile forms. Bacteria in the BMC medium grew more readily with more abundant metabolites than those raised in free cells, probably because biochar provides shelter via porous structures (as confirmed by scanning electron microscopy) as well as additional nutrients. Overall, the improved long-term production of microbial biomass caused by BMC inoculation results in a higher remediation efficiency. Our results demonstrate the feasibility of using biochar as an appropriate carrier for metal-tolerant bacteria to remediate Cd-contaminated paddy fields.

Enterobacter asburiae Reduces Cadmium Toxicity in Maize Plants by Repressing Iron Uptake-Associated Pathways.

Soil microbes have recently been utilized to improve cadmium (Cd) tolerance and lower its accumulation in plants. Nevertheless, whether rhizobacteria can prevent Cd uptake by graminaceous plants and the underlying mechanisms remain elusive. In this study, inoculation with Enterobacter asburiae NC16 reduced transpiration rates and the expression of some iron (Fe) uptake-related genes including ZmFer, ZmYS1, ZmZIP, and ZmNAS2 in maize (Zea mays) plants, which contributed to mitigation of Cd toxicity. However, the inoculation with NC16 failed to suppress the transpiration rates and transcription of these Fe uptake-related genes in plants treated with fluridone, an abscisic acid (ABA) biosynthetic inhibitor, indicating that the impacts of NC16-inoculation observed were dependent on the actions of ABA. We found that NC16 increased the host ABA levels by mediating the metabolism of ABA rather than its synthesis. Moreover, the capacity of NC16 to inhibit plant uptake of Cd was greatly weakened in plants overexpressing ZmZIP, encoding a zinc/iron transporter. Collectively, our findings indicated that E. asburiae NC16 reduced Cd toxicity in maize plants at least partially by hampering the Fe uptake-associated pathways.

Benzenedicarboxylic acid upregulates O48814 and Q9FJQ8 for improved nutritional contents of tomato and low risk of fungal attack.

BACKGROUND: Tomato is an important food item and a cocktail of phytonutrients. In the current study, metabolites from a non-pathogenic fungal species Penicillium oxalicum have been exploited to obtain nutritionally augmented tomato fruits from the plants to better withstand against Alternaria alternata infection. RESULTS: Initially, bioactivity-guided assay and chromatographic analyses identified the bioactive metabolites of P. oxalicum [benzenedicarboxylic acid (BDA) and benzimidazole]. Then, ≥3 times elevated quantities of vitamins and other nutritional elements (protein, fat, fibers, and carbohydrates) were achieved by the foliar application of BDA. The maximum increase (625.81%) was recorded in riboflavin contents; however, thiamine showed the second highest enhancement (542.86%). Plant metabolites analysis revealed that jasmonic acid contents were boosted 121.53% to significantly enhance guaiacyl lignin defenses along with the reduction in coumarin contents. The protein profile analysis explored three most actively responding protein species toward BDA applications, (i) palmitoyltransferase protein Q9FLM3; (ii) serine/threonine-protein kinase O48814; and (iii) E3 ubiquitin-protein ligase Q9FJQ8. The O48814 improved plant defenses; whereas, Q9FJQ8 protein was negatively regulating cysteine-type endopeptidase activity and assisted plant to resist schedule alterations. Tomato cultivar with more active innate metabolism was found to be more responsive toward BDA. Furthermore, the bioactive compounds were enriched by using the two-step extraction method of ethyl acetate and chloroform, respectively. CONCLUSION: Penicillium oxalicum a non-pathogenic fungal species, produced BDA, induced nutritional contents in tomato and protected it against Alternaria alternata. The current study is the first report on the bioactivity of BDA and benzimidazole concerning the nutritional enhancement and plant defense improvement. © 2019 Society of Chemical Industry.

Role of novel bacterial Raoultella sp. strain X13 in plant growth promotion and cadmium bioremediation in soil.

Heavy metal pollution in agricultural soils has become a widespread serious problem with the rapid industrialization and urbanization in the past two decades. Cadmium (Cd2+) is of the most concern in soils due to its high toxicity. It is necessary to develop remediation strategies to remove or neutralize its toxic effects in Cd-contaminated soil. Microbial bioremediation is a promising technology to treat heavy metal-contaminated soils. In this study, Cd-resistant bacterium, isolated from heavy metal-polluted soil in Southern China, was characterized as Raoultella sp. strain X13 on the basis of its biochemical profile and 16S rRNA. We investigated the characterization of Cd2+ distribution in different cellular compartments after Cd2+ uptake. Cd2+ uptake by strain X13 was mainly by ion exchange and chelation binding tightly to the cell wall. In addition, X13 plant growth-promoting characteristics suggested that X13 could solubilize phosphate and produce indole acetic acid. Pot experiments for the remediation of Cd-contaminated soil in situ by X13 inoculation demonstrated that X13 application to Cd-contaminated soils significantly promoted pak choi growth and improved production. We also found that X13 substantially reduced the Cd2+ bioavailability for pak choi. Therefore, strain X13 is an effective treatment for potential application in Cd2+ remediation as well as for sustainable agronomic production programs in Cd-contaminated soils.

Inoculation with Bacillus subtilis and Azospirillum brasilense Produces Abscisic Acid That Reduces Irt1-Mediated Cadmium Uptake of Roots.

Cadmium (Cd) contamination of agricultural soils represents a serious risk to crop safety. A new strategy using abscisic acid (ABA)-generating bacteria, Bacillus subtilis or Azospirillum brasilense, was developed to reduce the Cd accumulation in plants grown in Cd-contaminated soil. Inoculation with either bacterium resulted in a pronounced increase in the ABA level in wild-type Arabidopsis Col-0 plants, accompanied by a decrease in Cd levels in plant tissues, which mitigated the Cd toxicity. As a consequence, the growth of plants exposed to Cd was improved. Nevertheless, B. subtilis and A. brasilense inoculation had little effect on Cd levels and toxicity in the ABA-insensitive mutant snrk 2.2/2.3, indicating that the action of ABA is required for these bacteria to reduce Cd accumulation in plants. Furthermore, inoculation with either B. subtilis or A. brasilense downregulated the expression of IRT1 (iron-regulated transporter 1) in the roots of wild-type plants and had little effect on Cd levels in the IRT1-knockout mutants irt1-1 and irt1-2. In summary, we conclude that B. subtilis and A. brasilense can reduce Cd levels in plants via an IRT1-dependent ABA-mediated mechanism.

Promotion of iron nutrition and growth on peanut by Paenibacillus illinoisensis and Bacillus sp. strains in calcareous soil

ABSTRACT This study aimed to explore the effects of two siderophore-producing bacterial strains on iron absorption and plant growth of peanut in calcareous soil. Two siderophore-producing bacterial strains, namely, YZ29 and DZ13, isolated from the rhizosphere soil of peanut, were identified as Paenibacillus illinoisensis and Bacillus sp., respectively. In potted experiments, YZ29 and DZ13 enhanced root activity, chlorophyll and active iron content in leaves, total nitrogen, phosphorus and potassium accumulation of plants and increased the quality of peanut kernels and plant biomass over control. In the field trial, the inoculated treatments performed better than the controls, and the pod yields of the three treatments inoculated with YZ29, DZ13, and YZ29 + DZ13 (1:1) increased by 37.05%, 13.80% and 13.57%, respectively, compared with the control. Based on terminal restriction fragment length polymorphism analysis, YZ29 and DZ13 improved the bacterial community richness and species diversity of soil surrounding the peanut roots. Therefore, YZ29 and DZ13 can be used as candidate bacterial strains to relieve chlorosis of peanut and promote peanut growth. The present study is the first to explore the effect of siderophores produced by P. illinoisensis on iron absorption.

Promotion of iron nutrition and growth on peanut by Paenibacillus illinoisensis and Bacillus sp. strains in calcareous soil.

This study aimed to explore the effects of two siderophore-producing bacterial strains on iron absorption and plant growth of peanut in calcareous soil. Two siderophore-producing bacterial strains, namely, YZ29 and DZ13, isolated from the rhizosphere soil of peanut, were identified as Paenibacillus illinoisensis and Bacillus sp., respectively. In potted experiments, YZ29 and DZ13 enhanced root activity, chlorophyll and active iron content in leaves, total nitrogen, phosphorus and potassium accumulation of plants and increased the quality of peanut kernels and plant biomass over control. In the field trial, the inoculated treatments performed better than the controls, and the pod yields of the three treatments inoculated with YZ29, DZ13, and YZ29+DZ13 (1:1) increased by 37.05%, 13.80% and 13.57%, respectively, compared with the control. Based on terminal restriction fragment length polymorphism analysis, YZ29 and DZ13 improved the bacterial community richness and species diversity of soil surrounding the peanut roots. Therefore, YZ29 and DZ13 can be used as candidate bacterial strains to relieve chlorosis of peanut and promote peanut growth. The present study is the first to explore the effect of siderophores produced by P. illinoisensis on iron absorption.

Isolation and engineering of plant growth promoting rhizobacteria Pseudomonas aeruginosa for enhanced cadmium bioremediation.

Although many bacteria are tolerant to heavy metals and play important roles in the immobilization of heavy metals, they cannot always be dependably reproduced under field conditions. In this work, a cadmium (Cd)-resistant bacterium was isolated from a Cd-contaminated oil field and identified as Pseudomonas aeruginosa (Pse-w). We then determined various plant growth promoting features such as the solubilization of phosphate, and the production of indole-3-acetic acid and siderophores. Lastly, we engineered the strain Pse-w-MT by targeting metallothioneins to the cell surface of Pse-w to immobilize Cd2+ and promote plant growth. Our data revealed that Pse-w exhibited high levels of resistance to Cd2+ (4 mM) and showed various plant growth promoting features. The engineered strain Pse-w-MT was found to adsorb Cd2+ mainly via extracellular deposition, and had an enhanced ability for immobilizing Cd2+ ions from the external media. Furthermore, the inoculation of Cd-polluted soil with Pse-w-MT significantly elevated the shoot and root biomass and leaf chlorophyll content. Similarly, plants inoculated with Pse-w-MT demonstrated markedly lower Cd2+ accumulation in the root and shoot system. It was concluded that plant growth promoting rhizobacteria with a high Cd2+ tolerance was an ideal candidate to be engineered for bioremediation and plant growth promotion against Cd-induced stress.

Interrogantes en la tecnología de la inoculación de semillas de soja con Bradyrhizobium spp / Queries related to the technology of soybean seed inoculation with Bradyrhizobium spp

With the aim of exploiting symbiotic nitrogen fixation, soybean crops are inoculated with selected strains of Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens or Bradyrhizobium elkanii (collectively referred to as Bradyrhizobium spp.). The most common method of inoculation used is seed inoculation, whether performed immediately before sowing or using preinoculated seeds or pretreated seeds by the professional seed treatment. The methodology of inoculation should not only cover the seeds with living rhizobia, but must also optimize the chances of these rhizobia to infect the roots and nodulate. To this end, inoculated rhizobia must be in such an amount and condition that would allow them to overcome the competition exerted by the rhizobia of the allochthonous population of the soil, which are usually less effective for nitrogen fixation and thus dilute the effect of inoculation on yield. This optimization requires solving some queries related to the current knowledge of seed inoculation, which are addressed in this article. I conclude that the aspects that require further research are the adhesion and survival of rhizobia on seeds, the release of rhizobia once the seeds are deposited in the soil, and the movement of rhizobia from the vicinity of the seeds to the infection sites in the roots

Con el fin de aprovechar la fijación simbiótica de nitrógeno, el cultivo de soja se inocula con cepas seleccionadas de Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens o Bradyrhizobium elkanii (conjuntamente referidas como Bradyrhizobium spp.). El método más común de hacerlo es la inoculación en semillas, ya sea que esta se realice en el momento previo a la siembra o que se utilicen semillas preinoculadas o pretratadas mediante el tratamiento profesional de semillas. La metodología de inoculación no debe limitarse a recubrir las semillas con rizobios vivos, sino que también debe optimizar las chances de esos rizobios para infectar las raíces y nodular. Para ello los rizobios inoculados deben estar en una cantidad y un estado tales que les permitan superar la competición ejercida por los rizobios de la población alóctona del suelo, los cuales usualmente son menos eficaces para la fijación de nitrógeno y así diluyen el efecto de la inoculación sobre el rendimiento. Esta optimización requiere resolver algunos interrogantes, que son abordados en el presente artículo. Concluyo que los aspectos que requieren más investigación son la adhesión y supervivencia de los rizobios en las semillas, la liberación de los rizobios una vez que las semillas se depositan en el suelo y el movimiento de los rizobios desde las inmediaciones de las semillas hasta los sitios de infección en las raíces

Effects of biodegradation and sorption by humic acid on the estrogenicity of 17ß-estradiol.

The removal of 17ß-estradiol (E2) by biodegradation and sorption onto humic acid (HA) was examined at various HA concentrations. Subsequently, estrogenicity associated with E2 removal was estimated using E-screen bioassay. Results showed that E2 biodegradation and its subsequent transformation to estrone (E1) were significantly reduced with increasing HA concentration. In addition, the presence of nutrients enhanced the biodegradation of E2. Overall, E2 biodegradation was the dominating contributor to its removal, which demonstrated a significantly negative correlation with E2 sorption at various HA concentrations. The sorption of E2 by HA was significantly enhanced with increasing HA concentration. Estrogenicity associated with residual E2 showed that there existed a significant difference among various HA concentrations, with the lowest value in the absence of HA. The findings suggest that the presence of HA and nutrients in natural waters should be considered in assessing estrogenicity of environmental samples due to complex sorption and biodegradation processes.