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Although arbuscular mycorrhizal (AM) fungi are known to promote growth and yield of agricultural crops, inoculation methods for effective scaling up from greenhouse to the field are still underexplored. The application of single or mixed beneficial AM fungal isolates is hindered by the lack of experimental reproducibility of findings at different scales and the cost-effectivity of inoculation methods. Seed coating has been considered a feasible delivery system of AM fungal inocula for agricultural crops. In this study, the impact of single and multiple AM fungal isolates applied via seed coating on chickpea productivity was evaluated under greenhouse and field conditions. Overall, plants inoculated with multiple AM fungal isolates had better performance than those inoculated with single AM isolate under greenhouse and field conditions. While plants in greenhouse displayed higher shoot dry weight (14%) and seed individual weight (21%), in field, inoculation with multiple AM isolates increased pod (160%), and seed (148%) numbers, and grain yield (140%). Under field conditions, mycorrhizal root colonization was significantly higher in chickpea plants inoculated with multiple AM fungal isolates compared to other treatments. These findings highlight the potential of field-inoculation with multiple AM fungal isolates via seed coating as a sustainable agricultural practice for chickpea production.

Phytoremediation has been considered as a promising technique to decontaminate polluted soils. However, climatic stress particularly salinity, is a potential threat to soil properties and plant growth, thus restricting the employment of this technology. The aim of this study was to access the impact of microbial inoculation on phytoremediation of nickel (Ni) contaminated saline soils using Helianthus annuus. Salt resistant plant beneficial bacterium (PBB) Pseudomonas libanensis TR1 and arbuscular mycorrhizal fungus (AMF) Claroideoglomus claroideum BEG210 were used. Inoculation of P. libanensis alone or in combination with C. claroideum significantly enhanced plant growth, changed physiological status (e.g. electrolyte leakage, chlorophyll, proline and malondialdehyde contents) as well as Ni and sodium (Na+) accumulation potential (e.g. uptake and translocation factor of Ni and Na+) of H. annuus under Ni and salinity stress either alone or in combination. These results revealed that bioaugmentation of microbial strains may serve as a preferred strategy for improving phytoremediation of metal-polluted saline soils.

Cowpea (Vigna unguiculata L. Walp) is an important legume grown primarily in semi-arid area. Its production is generally inhibited by various abiotic and biotic stresses. The use of beneficial microorganisms (e.g., plant growth promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF)) can enhance agricultural production, as these microorganisms can improve soil fertility and plant tolerance to environmental stresses, thus enhancing crop yield in an eco-friendly manner. Application of PGPB and AMF in large scale agriculture needs to be improved. Thus, the use of seed coating could be an efficient mechanism for placement of inocula into soils. The aim of this study was to evaluate the effects of the AMF Rhizophagus irregularis BEG140 and the PGPB Pseudomonas libanensis TR1 alone or in combination on the biomass and physiological traits of cowpea. Four treatments were set: (i) non-inoculated control, (ii) PGPB, (iii) AMF applied via seed coating, and (iv) PGPB + AMF applied via seed coating. Cowpea plants inoculated via seed coating with R. irregularis and those inoculated with R. irregularis + P. libanensis showed root mycorrhizal colonization of 21.7% and 24.2%, respectively. PGPB P. libanensis was efficient in enhancing plant biomass and seed yield. There was no benefit of single (AMF) or dual (PGPB + AMF) inoculation on plant growth or seed yield. The application of beneficial soil microorganisms can be a viable approach for sustainable cowpea production in precision agriculture scenarios.

A plant growth-promoting bacterial (PGPB) strain SC2b was isolated from the rhizosphere of Sedum plumbizincicola grown in lead (Pb)/zinc (Zn) mine soils and characterized as Bacillus sp. based on (1) morphological and biochemical characteristics and (2) partial 16S ribosomal DNA sequencing analysis. Strain SC2b exhibited high levels of resistance to cadmium (Cd) (300 mg/L), Zn (730 mg/L), and Pb (1400 mg/L). This strain also showed various plant growth-promoting (PGP) features such as utilization of 1-aminocyclopropane-1-carboxylate, solubilization of phosphate, and production of indole-3-acetic acid and siderophore. The strain mobilized high concentration of heavy metals from soils and exhibited different biosorption capacity toward the tested metal ions. Strain SC2b was further assessed for PGP activity by phytagar assay with a model plant Brassica napus. Inoculation of SC2b increased the biomass and vigor index of B. napus. Considering such potential, a pot experiment was conducted to assess the effects of inoculating the metal-resistant PGPB SC2b on growth and uptake of Cd, Zn and Pb by S. plumbizincicola in metal-contaminated agricultural soils. Inoculation with SC2b elevated the shoot and root biomass and leaf chlorophyll content of S. plumbizincicola. Similarly, plants inoculated with SC2b demonstrated markedly higher Cd and Zn accumulation in the root and shoot system, indicating that SC2b enhanced Cd and Zn uptake by S. plumbizincicola through metal mobilization or plant-microbial mediated changes in chemical or biological soil properties. Data demonstrated that the PGPB Bacillus sp. SC2b might serve as a future biofertilizer and an effective metal mobilizing bioinoculant for rhizoremediation of metal polluted soils.

This work focused on the biodegradation of three structurally related fluoroacetates (FAs), mono- (MFA), di- (DFA) and trifluoroacetate (TFA), using as microbial inocula samples collected from a site with a long history of industrial contamination and activated sludge obtained from a municipal wastewater treatment plant. Biodegradation experiments were carried out under different modes of substrate supplementation, which included (i) FAs fed as sole carbon sources; (ii) FAs (only for DFA and TFA) fed in co-metabolism with sodium acetate; and (iii) mixtures of MFA with DFA or TFA. Biodegradation of the target compounds was assessed through fluoride ion release. Defluorination was obtained in the cultures fed with MFA, while DFA and TFA were recalcitrant in all tested conditions. When present in mixture, DFA was shown to inhibit biodegradation of MFA, while TFA had no effect. A total of 13 bacterial isolates obtained from MFA degrading cultures were found to degrade 20 mgL −1 of this compound, as single strains, when supplemented as a sole carbon source. Sequencing of the 16S rRNA gene indicated that among these degrading bacteria only Delftia acidovorans had been previously reported to be able to degrade MFA. This work shows that, despite their similar chemical structures, biodegradation of the three tested FAs is very distinct and draws attention to the unknown impacts that the accumulation of DFA and TFA may have in the environment as a result of their high recalcitrance. © 2017 Elsevier R.S. Oliveira wishes to acknowledge the support of Fundação para a Ciência e a Tecnologia (FCT) through the research grant SFRH/BPD/85008/2012 and Fundo Social Europeu (FSE) . M.F. Carvalho and F. Pereira acknowledge Investigator FCT program supported by FCT , FSE and Programa Operacional Potencial Humano . This work was financed by national funds through FCT within the scope of the project IF/00791/2013/CP1197/CT0002 .

Diflubenzuron (DFB) is an insecticide commonly used to control forest pests. The objectives of this study were to assess the effect of diflubenzuron on the development of Pinus pinaster seedlings and Pisolithus tinctorius under laboratory conditions and to study the possible protective role of this ectomycorrhizal fungus against the effects of diflubenzuron. In vitro experiments revealed that diflubenzuron inhibited fungal growth at all tested concentrations (0.01, 0.1, 1, 10 and 100 mg L(-1)). Root growth was inhibited at the two highest diflubenzuron concentrations. The activity of the antioxidant defence system of non-inoculated P. pinaster increased at 1 and 10 mg DFB kg(-1) substrate, and inoculation increased the threshold to the highest concentration. The protective role of the ectomycorrhizal fungus was seen in the increase of CAT activity. This study revealed that despite causing no mortality, diflubenzuron has the ability to cause sub-lethal damage to P. pinaster. The disproportionate use of this insecticide may lead to higher amounts of its residues in soil and the biosphere, endangering trees, fungi and their symbiosis.

Plants and microbes coexist or compete for survival and their cohesive interactions play a vital role in adapting to metalliferous environments, and can thus be explored to improve microbe-assisted phytoremediation. Plant root exudates are useful nutrient and energy sources for soil microorganisms, with whom they establish intricate communication systems. Some beneficial bacteria and fungi, acting as plant growth promoting microorganisms (PGPMs), may alleviate metal phytotoxicity and stimulate plant growth indirectly via the induction of defense mechanisms against phytopathogens, and/or directly through the solubilization of mineral nutrients (nitrogen, phosphate, potassium, iron, etc.), production of plant growth promoting substances (e.g., phytohormones), and secretion of specific enzymes (e.g., 1-aminocyclopropane- 1-carboxylate deaminase). PGPM can also change metal bioavailability in soil through various mechanisms such as acidification, precipitation, chelation, complexation, and redox reactions. This review presents the recent advances and applications made hitherto in understanding the biochemical and molecular mechanisms of plant–microbe interactions and their role in the major processes involved in phytoremediation, such as heavy metal detoxification, mobilization, immobilization, transformation, transport, and distribution.

Fluorinated compounds are finding increasing uses in several applications. They are employed in almost all areas of modern society. These compounds are all produced by chemical synthesis and their abundance highly contrasts with fluorinated molecules of natural origin. To date, only some plants and a handful of actinomycetes species are known to produce a small number of fluorinated compounds that include fluoroacetate (FA), some ω-fluorinated fatty acids, nucleocidin, 4-fluorothreonine (4-FT), and the more recently identified (2R3S4S)-5-fluoro-2,3,4-trihydroxypentanoic acid. This largely differs from other naturally produced halogenated compounds, which totals more than 5000. The mechanisms underlying biological fluorination have been uncovered after discovering the first actinomycete species, Streptomyces cattleya, that is capable of producing FA and 4-FT, and a fluorinase has been identified as the enzyme responsible for the formation of the C–F bond. The discovery of this enzyme has opened new perspectives for the biotechnological production of fluorinated compounds and many advancements have been achieved in its application mainly as a biocatalyst for the synthesis of [18F]-labeled radiotracers for medical imaging. Natural fluorinated compounds may also be derived from abiogenic sources, such as volcanoes and rocks, though their concentrations and production mechanisms are not well known. This review provides an outlook of what is currently known about fluorinated compounds with natural origin. The paucity of these compounds and the biological mechanisms responsible for their production are addressed. Due to its relevance, special emphasis is given to the discovery, characterization and biotechnological potential of the unique fluorinase enzyme. © 2016 Informa UK Limited, trading as Taylor & Francis Group M. F. Carvalho acknowledges Investigator FCT program supported by Fundação para a Ciência e a Tecnologia (FCT), Fundo Social Europeu (FSE), and Programa Operacional Potencial Humano. R. S. Oliveira wishes to acknowledge the support of FCT through the research grant SFRH/BPD/85008/2012, FSE and Programa Operacional Capital Humano.

Zn tissue accumulation in Solanum nigrum grown in a non-contaminated and a naturally contaminated Zn matrix and the effect of inoculation with different arbuscular mycorrhizal fungi (AMF) on metal uptake were assessed. S. nigrum grown in the contaminated soil always presented higher Zn accumulation in the tissues, accumulating up to 1622 mg Zn kg −1 . The presence of both Glomus claroideum and Glomus intraradices enhanced the uptake and accumulation of Zn by S. nigrum (up to 83 and 49% higher Zn accumulation, respectively). The main deposits of the metal were found in the intercellular spaces and in the cell walls of the root tissues, as revealed by autometallography, with the inoculation with different AMF species causing no differences in the location of Zn accumulation. These findings indicate that S. nigrum inoculated with selected heavy metal tolerant AMF presents extracting and accumulating capacities, constituting a potentially suitable remediation method for Zn polluted soils.

Purpose: Polycyclic aromatic hydrocarbons (PAHs) are a class of organic compounds commonly found as soil contaminants. Fungal degradation is considered as an environmentally friendly and cost-effective approach to remove PAHs from soil. Acenaphthylene (Ace) and Benzo[a]anthracene (BaA) are two PAHs that can coexist in soils; however, the influence of the presence of each other on their biodegradation has not been studied. The biodegradation of Ace and BaA, alone and in mixtures, by the white rot fungus Pleurotus ostreatus was studied in a sandy soil. Materials and methods: Experimental microcosms containing soil spiked with different concentrations of Ace and BaA were inoculated with P. ostreatus. Initial (t 0) and final (after 15 days of incubation) soil concentrations of Ace and BaA were determined after extraction of the PAHs. Results and discussion: P. ostreatus was able to degrade 57.7% of the Ace in soil spiked at 30 mg kg-1 dry soil and 65.8% of Ace in soil spiked at 60 mg kg-1 dry soil. The degradation efficiency of BaA by P. ostreatus was 86.7 and 77.4% in soil spiked with Ace at 30 and 60 mg kg-1 dry soil, respectively. After 15 days of incubation, there were no significant differences in Ace concentration between soil spiked with Ace and soil spiked with Ace + BaA, irrespective of the initial soil concentration of both PAHs. There were also no differences in BaA concentration between soil spiked with BaA and soil spiked with BaA + Ace. Conclusions: The results indicate that the fungal degradation of Ace and BaA was not influenced by the presence of each other's PAH in sandy soil. Bioremediation of soils contaminated with Ace and BaA using P. ostreatus is a promising approach to eliminate these PAHs from the environment.

The effect of two different chelating agents [EDTA and EDDS (S,S-ethylenediaminedissucinic acid)] on Zn tissue accumulation in Solanum nigrum L. grown in a naturally contaminated soil was assessed. Under those conditions, the response of the plant to the inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF) - Glomus claroideum and Glomus intraradices - was also studied. Plants grown in the local contaminated soil (Zn levels of 433 mg kg-1) accumulated up to 1191 mg kg-1 of Zn in the roots, 3747 mg kg-1 in the stems and 3409 mg kg-1 in the leaves. S. nigrum plants grown in the same soil spiked with extra Zn (Zn levels of 964 mg kg-1) accumulated up to 4735, 8267 and 7948 mg Zn kg-1 in the leaves, stems and roots, respectively. The addition of EDTA promoted an increase in the concentration of Zn accumulated by S. nigrum of up to 231% in the leaves, 93% in the stems and 81% in the roots, while EDDS application enhanced the accumulation in leaves, stems and roots up to 140, 124 and 104%, respectively. In the stems, the presence of Zn was predominantly detected in the cortex collenchyma cells, the starch sheath and the internal phloem and xylem parenchyma, and the addition of chelating agents did not seem to have an effect on the localisation of accumulation sites. The devise of a chelate-enhanced phytoextraction strategy, using chelating agents and AMF, is discussed.

Drought can drastically reduce cowpea [Vigna unguiculata (L.) Walp.] biomass and grain yield. The application of plant growth‐promoting rhizobacteria and arbuscular mycorrhizal fungi can confer resistance to plants and reduce the effects of environmental stresses, including drought. Seed coating is a technique which allows the application of minor amounts of microbial inocula. Main effects of the factors inoculation and water regime showed that: severe or moderate water deficit had a general negative impact on cowpea plants; total biomass production, seed weight and seed yield were enhanced in plants inoculated with P. putida; inoculation of R. irregularis significantly increased nitrogen (N) and phosphorus (P) shoot concentrations; and R. irregularis enhanced both chlorophyll b and carotenoids contents, particularly under severe water deficit. Plants inoculated with P. putida + R. irregularis had an increase in shoot P concentration of 85% and 57%, under moderate and severe water deficit, respectively. Singly inoculated P. putida improved potassium shoot concentration by 25% under moderate water deficit. Overall, in terms of agricultural productivity the inoculation of P. putida under water deficit might be promising. Seed coating has the potential to be used as a large‐scale delivery system of beneficial microbial inoculants.