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Bacteria produce a large number of volatile organic compounds (VOCs) into the environment. VOCs are species-specific and their emission depends on environmental conditions, such as growth medium, pH, temperature, incubation time and interaction with other microorganisms. These VOCs can enhance plant growth, suppress pathogens and act as signaling molecules during plant-microorganism interactions. Bacterial VOCs have been reported to show strong antimicrobial, nematicidal, pesticidal, plant defense, induced tolerance and plant-growth-promoting activities under controlled conditions. Commonly produced antifungal VOCs include dimethyl trisulfide, dimethyl disulfide, benzothiazole, nonane, decanone and 1-butanol. Species of Bacillus, Pseudomonas, Arthrobacter, Enterobacter and Burkholderia produce plant growth promoting VOCs, such as acetoin and 2,3-butenediol. These VOCs affect genes involved in defense and development in plant species (i.e., Arabidopsis, tobacco, tomato, potato, millet and maize). VOCs are also implicated in altering pathogenesis-related genes, inducing systemic resistance, modulating plant metabolic pathways and acquiring nutrients. However, detailed mechanisms of action of VOCs need to be explored. This review summarizes the bioactive VOCs produced by diverse bacterial species as an alternative to agrochemicals, their mechanism of action and challenges for employment of bacterial VOCs for sustainable agricultural practices. Future studies on technological improvements for bacterial VOC application under greenhouse and open field conditions are warranted.

Elevated CO2 stimulates plant growth and affects quantity and composition of root exudates, followed by response of its microbiome. Three scenarios representing nitrate fertilization regimes: limited (30 ppm), moderate (70 ppm) and excess nitrate (100 ppm) were compared under ambient and elevated CO2 (eCO2, 850 ppm) to elucidate their combined effects on root-surface-associated bacterial community abundance, structure and function. Wheat root-surface-associated microbiome structure and function, as well as soil and plant properties, were highly influenced by interactions between CO2 and nitrate levels. Relative abundance of total bacteria per plant increased at eCO2 under excess nitrate. Elevated CO2 significantly influenced the abundance of genes encoding enzymes, transporters and secretion systems. Proteobacteria, the largest taxonomic group in wheat roots (~ 75%), is the most influenced group by eCO2 under all nitrate levels. Rhizobiales, Burkholderiales and Pseudomonadales are responsible for most of these functional changes. A correlation was observed among the five gene-groups whose abundance was significantly changed (secretion systems, particularly type VI secretion system, biofilm formation, pyruvate, fructose and mannose metabolism). These changes in bacterial abundance and gene functions may be the result of alteration in root exudation at eCO2, leading to changes in bacteria colonization patterns and influencing their fitness and proliferation.

Microbial communities are highly responsive to environmental cues, and both their structure and activity can be altered in response to changing conditions. We hypothesized that host-associated microbial communities, particularly those colonizing host surfaces, can serve asin situsensors to reveal environmental conditions experienced by both microorganisms and the host. For a proof-of-concept, we studied a model plant-soil system and employed a non-deterministic gene-centric approach. A holistic analysis was performed using plants of two species and irrigation with water of low quality to induce host stress. Our analyses examined the genetic potential (DNA) and gene expression patterns (RNA) of plant-associated microbial communities, as well as transcriptional profiling of host plants. Transcriptional analysis of plants irrigated with treated wastewater revealed significant enrichment of general stress-associated root transcripts relative to plants irrigated with fresh water. Metagenomic analysis of root-associated microbial communities in treated wastewater-irrigated plants, however, revealed enrichment of more specific stress-associated genes relating to high levels of salt, high pH and lower levels of oxygen. Meta-analysis of these differentially abundant genes obtained from other metagenome studies provided evidence of the link between environmental factors such as pH and oxygen and these genes. Analysis of microbial transcriptional response demonstrated that enriched gene content was actively expressed, which implies contemporary response to elevated levels of pH and salt. We demonstrate here that microbial profiling can elucidate stress signals that cannot be observed even through interrogation of host transcriptome, leading to an alternate mechanism for evaluatingin situconditions experienced by host organisms.Significance StatementThis study examines the potential for microbial communities to provide insight into stresses experienced by their eukaryotic host organisms, through profiling of metagenomes and metatranscriptomes. Our study uses plant host-associated microorganisms as anin vivoandin situmicrosensor to identify environmental stresses experienced by the microbial community and by the plant. Transcriptionally active host-associated microbial communities are responsive in a highly specific manner to environmental conditions. Conversely, host transcriptome sequencing provides only a very general stress response. This study is a proof-of-concept for the use of microbial communities as microsensors, with a great potential for interrogation of a wide range of host systems.

Root selection of their associated microbiome composition and activities is determined by the plant’s developmental stage and distance from the root. Total gene abundance, structure and functions of root-associated and rhizospheric microbiomes were studied throughout wheat growth season under field conditions. On the root surface, abundance of the well-known wheat colonizers Proteobacteria and Actinobacteria decreased and increased, respectively, during spike formation, whereas abundance of Bacteroidetes was independent of spike formation. Metagenomic analysis combined with functional co-occurrence networks revealed a significant impact of plant developmental stage on its microbiome during the transition from vegetative growth to spike formation. For example, gene functions related to biofilm and sensorial movement, antibiotic production and resistance and carbons and amino acids and their transporters. Genes associated with these functions were also in higher abundance in root vs. the rhizosphere microbiome. We propose that abundance of transporter-encoding genes related to carbon and amino acid, may mirror the availability and utilization of root exudates. Genes related to antibiotic resistance mechanisms were abundant during vegetative growth, while after spike formation, genes related to the biosynthesis of various antibiotics were enriched. This observation suggests that during root colonization and biofilm formation, bacteria cope with competitor’s antibiotics, whereas in the mature biofilm stage, they invest in inhibiting new colonizers. Additionally, there is higher abundance of genes related to denitrification in rhizosphere compared to root-associated microbiome during wheat growth, possibly due to competition with the plant over nitrogen in the root vicinity. We demonstrated functional and phylogenetic division in wheat root zone microbiome in both time and space: pre- and post-spike formation, and root-associated vs. rhizospheric niches. These findings shed light on the dynamics of plant–microbe and microbe–microbe interactions in the developing root zone.

With increasing fresh water (FW) scarcity, the use of treated wastewater (TWW) for crop irrigation is expanding globally. Besides clear benefits, some undesired long-term effects of irrigation with this low quality water on plant performance have been reported. As the rhizosphere microbiome can mediate plant-soil interactions, an examination of the response of these organisms to TWW is necessary to understand the full effects of water quality. In the current study, the effects of irrigation water quality on the microbial community structure of soil and roots as well as edaphic properties and plant performance were evaluated. We compared soil and roots microbiomes of two different plant species (tomato and lettuce), each grown in two distinct soils, and irrigated with either FW or TWW. Irrigation with TWW significantly increase soil pH, EC, K, Na and DOC, and decrease plant fruit and shoot weight, relatively to samples irrigated with FW. We calculated the effect size of plant species, soil type, and irrigation water quality on microbial community structure in soil and root. In the roots, plant species and irrigation water were the dominant factors in shaping both total (DNA based) and active (RNA based) microbial communities, with both factors contributing similarly to the observed microbial population. Soil type and irrigation water were the dominant factors shaping the total microbial community in the soil and were of similar magnitude. Irrigation water quality is demonstrated to be a major force in shaping root-associated microbiome, leading to altered microbial community structure in the critical juncture between plant and soil.

Background Roots select their associated microbiome and affect its composition and activities through exudates that provide a nutrient-rich environment based on distance from the root. Root-exudation patterns depend on the plant's developmental stage. We followed field-grown wheat from emergence to spike maturation and compared the structure and functions of the microbiomes in two niches: adjacent, root-associated bacteria and the rhizosphere. The effects of growth stage on root-associated and rhizospheric communities structure and functions were investigated to enhance our understanding of plant–microbe interactions. Results A significant impact of wheat developmental stage during the transition from vegetative growth to spike formation was observed on structure and functions of both root-associated and rhizosphere microbiomes. On the root surface, abundance of the well-known wheat colonizers Proteobacteria and Actinobacteria decreased and increased, respectively, during spike formation, whereas abundance of Bacteroidetes was independent of spike formation. Three microbiome functional clusters were specifically associated with root proximity: (1) biofilm and sensorial movement; (2) antibiotic production and resistance; and (3) carbon biosynthesis, degradation and transporters, and amino acid biosynthesis and transporters. Interestingly, in the root-associated microbiome, genes related to all of these functions were influenced by spike formation. Abundance of genes related to nine other functions (lipopolysaccharide transporter and biosynthesis, beta-lactam resistance, metabolism of methane, alanine-aspartate-glutamate and arginine-proline, and biosynthesis of peptidoglycan, lysine and enediyne antibiotics) was significantly influenced by spike formation in both roots and rhizosphere. All of these genes were abundant in the rhizosphere, more so before spike formation when root exudation is high, supporting the notion that some of the root exudates are not fully utilized by the root-associated bacteria and subsequently diffuse into the surrounding soil, creating the rhizosphere. Conclusions We demonstrated functional division in the microbiome of the wheat root zone in both time and space: pre- and post-spike formation and root-associated vs. rhizospheric niches. The responses of the root microbiome were driven by both the plant and the microbial physiology and activities, both of which respond to environmental cues. These findings shed light on the dynamics of plant–microbe and microbe–microbe interactions in the root zone.

Excessive use of antimicrobials in aquaculture is concerning, given possible environmental ramifications and the potential contribution to the spread of antimicrobial resistance (AR). In this study, we explored seasonal abundance of antimicrobial resistance genes and bacterial community composition in the water column of an intensive aquaculture pond stocked with Silver Carp (Hypophthalmichthys molitrix) prophylactically treated with sulfamethoprim (25% sulfadiazine, 5% trimethoprim), relative to an adjacent unstocked reservoir. Bacterial community composition was monitored using high-throughput sequencing of 16S rRNA gene amplicons in eight sampling profiles to determine seasonal dynamics, representing principal stages in the fish fattening cycle. In tandem, qPCR was applied to assess relative abundance of selected antimicrobial resistance genes (sul1, sul2, dfrA1, tetA and blaTEM) and class-1 integrons (int1). Concomitantly, resistomes were extrapolated from shotgun metagenomes in representative profiles. Analyses revealed increased relative abundance of sulfonamide and tetracycline resistance genes in fishpond-03, relative to pre-stocking and reservoir levels, whereas no significant differences were observed for genes encoding resistance to antimicrobials that were not used in the fishpond-03. Seasons strongly dictated bacterial community composition, with high abundance of cyanobacteria in summer and increased relative abundance of Flavobacterium in the winter. Our results indicate that prophylactic use of sulfonamides in intensive aquaculture ponds facilitates resistance suggesting that prophylactic use of these antimicrobials in aquaculture should be restricted.

Ultraviolet (UV) irradiation is an emerging strategy for controlling the formation of undesired biofilms in water desalination facilities using reverse osmosis (RO). However, most studies examining these pretreatments are limited as they have been conducted on single-species cultures, while biofilms are composed of multiple-species communities. The goal of this study was to investigate the effect of UV-C irradiation on a model community composed of six environmental isolates from a marine biofilm formed in RO seawater desalination plant. There was a high variance in the susceptibility of the single-isolate cultures to UV-C, from no response (isolate Eryth23) to complete inactivation (isolate Vib3). The most active wavelength was around 260 nm, resulting in a loss of viability of single-isolate cultures and loss of vitality of the mixed-isolate cultures. With respect to biofilm formation, the activity of this wavelength was completely different compared to its activity on planktonic suspension. Irradiation with 260 nm did not inhibit the total biofilm formation by the six-isolate culture; moreover, isolates such as the resistant Eryth23 or the susceptible Pseudoalt17, even gained abundance in the mixed isolate biofilm. The only decrease in total biofilm was obtained from irradiation at 280 nm, which was less active against the planktonic culture. These results indicate that the complexity of the biofilm-forming microbial community may contribute to its resistance to UV-C irradiation. Significance and impact of the study This study examined the resistance of a multiple-isolate native marine culture to UV-C irradiation, in terms of viability, vitality and the ability to form biofilm. Results of this study showed that even though most of the cells were inactivated both in single-isolate and in multiple-isolate cultures, still the multiple-isolate cultures manages to form biofilms, surprisingly with higher biomass than without irradiation. The significance of the study is in its conclusion that studies on UV-C irradiation of biofilm-forming model micro-organisms are not always applicable to natural multiple-species communities.

Soil solarization, a method of chemical-free pest treatment, is a practical and cost-effective way to treat organic farming soil. This method uses polyethylene sheets to capture solar irradiation that heats the soil. Together with heat generated during composting of fresh organic matter, this method can effectively treat many soil-borne pathogens. In this study we examined the effects of solarization with the addition of compost at three maturation levels, on soil microbial community structure and function along with plant performance. Similar hydrolytic activity among the treatments (as measured by FDA hydrolysis) was measured in the first year of solarization. High level of activity was obtained in soil amended with immature compost at the beginning of the second year, suggesting residual (carry-over) effects of previous-year treatments. Amplicon sequencing of 16S rRNA encoding genes was used to study soil bacterial community structure. Diversity and richness of bacterial communities were found to be negatively affected by solarization in all treatments. Interestingly, bacterial communities of solarized soils clustered together, regardless of compost amended and type, separated from the cluster of communities from non-solarized soils of all amendments. Specifically, long and short terms negative effects of solarization on the relative abundance of Alphaproteobacteria, Acidobacteria and Actinobacteria were noticed. In contrast, solarization positively affected Bacilli and Gammaproteobacteria abundance in both long and short term. Furthermore, solarization had long and short term positive effects on productivity of eggplant and wheat plants. This study is the first to describe in high details the combined effects of solarization and amendment of composts of different maturation levels.

Nitrous oxide (N2O) is a greenhouse gas and a potent ozone-depleting substance in the stratosphere. Agricultural soils are one of the main global sources of N2O emissions, particularly from cereal fields due to their high areal coverage. The aim of this study was to isolate N2O-reducing bacteria able to mitigate N2O emissions from the soil after inoculation. We isolated several bacteria from wheat roots that were capable of N2O reduction in vitro and studied their genetic potential and activity under different environmental conditions. Three of these isolates- all carrying the nitrous oxide reductase-encoding clade I nosZ, able to reduce N2O in vitro, and efficient colonizers of wheat roots- presented different N2O-reduction strategies when growing in the root zone, possibly due to the different conditions in situ and their metabolic preferences. Each isolate seemed to prefer to operate at different altered oxygen levels. Isolate AU243 (related to Agrobacterium/Rhizobium) could reduce both nitrate and N2O and operated better at lower oxygen levels. Isolate AU14 (related to Alcaligenes faecalis), lacking nitrate reductases, operated better under less anoxic conditions. Isolate NT128 (related to Pseudomonas stutzeri) caused slightly increased N2O emissions under both anoxic and ambient conditions. These results therefore emphasize the importance of a deep understanding of soil–plant–microbe interactions when environmental application is being considered.

The use of biostimulants with plant growth-promoting properties, but without significant input of nutrients, is discussed as a strategy to increase stress resistance and nutrient use efficiency of crops. However, limited reproducibility under real production conditions remains a major challenge. The use of combination products based on microbial and non-microbial biostimulants or microbial consortia, with the aim to exploit complementary or synergistic interactions and increase the flexibility of responses under different environmental conditions, is discussed as a potential strategy to overcome this problem. This study aimed at comparing the efficiency of selected microbial single-strain inoculants with proven plant-growth promoting potential versus consortium products under real production conditions in large-scale tomato cultivation systems, exposed to different environmental challenges. In a protected greenhouse production system at Timisoara, Romania, with composted cow manure, guano, hair-, and feather-meals as major fertilizers, different fungal and bacterial single-strain inoculants, as well as microbial consortium products, showed very similar beneficial responses. Nursery performance, fruit setting, fruit size distribution, seasonal yield share, and cumulative yield (39&ndash, 84% as compared to the control) were significantly improved over two growing periods. By contrast, superior performance of the microbial consortia products (MCPs) was recorded under more challenging environmental conditions in an open-field drip-fertigated tomato production system in the Negev desert, Israel with mineral fertilization on a high pH (7.9), low fertility, and sandy soil. This was reflected by improved phosphate (P) acquisition, a stimulation of vegetative shoot biomass production and increased final fruit yield under conditions of limited P supply. Moreover, MCP inoculation was associated with selective changes of the rhizosphere-bacterial community structure particularly with respect to Sphingobacteriia and Flavobacteria, reported as salinity indicators and drought stress protectants. Phosphate limitation reduced the diversity of bacterial populations at the root surface (rhizoplane) and this effect was reverted by MCP inoculation, reflecting the improved P status of the plants. The results support the hypothesis that the use of microbial consortia can increase the efficiency and reproducibility of BS-assisted strategies for crop production, particularly under challenging environmental conditions.

Effect of temperature on the survival of Macrophomina phaseolina s.l. (Mp) was tested under laboratory conditions using an incubation system programmed for a cyclic temperature regime, simulating soil solarization. Mp culture and soil with and without Mp were incubated under two cyclic temperature regimes, with maximum temperatures of 50 °C or 52 °C, simulating solarization processes, and at a constant temperature of 25 °C, as a control. Total fungal community and Mp levels were not affected negatively by constant temperature of 25 °C. However, total soil fungal community was significantly decreased at temperature regime of 50 °C, whereas Mp did not change significantly whether in soil mixture or as pure propagules culture. In Mp-amended soil treated by temperature regime of 50 °C, total fungal community decreased while Mp CFU number did not change significantly. However, temperature regime of 52 °C almost completely eradicated total fungal community, as well as that of Mp. When Mp was amended to soils after being treated at 50 °C and 52 °C temperature regimes and incubated for two additional weeks at 25 °C, an indirect temperature effects on Mp survival were apparent. The 50 °C pre-treatment did affect significantly neither the total fungal nor Mp levels. In soil previously treated by 52 °C, total fungal growth was somewhat decreased while Mp growth was promoted. The results suggest that Mp survival may be affected by temperature directly as well as indirectly via effects on total fungal population. Studying these biological factors may contribute to the control of Mp pathogen using environment-friendly methods.

The combined effects of soil solarization and application of compost of various maturity levels upon soil microbial activity were studied under field conditions during 2010–2012. Eight treatments were divided into solarized and non-solarized treatments, and each was either non-amended or annually amended with compost at 6 kg (DW) m-2. The composts were mature, partially mature, or immature. In all three tested parameters of microbial activity (respiration rate, heat output, and dehydrogenase activity) the immature compost showed higher activity than the other compost types. Soil samples were collected weekly in order to assess microbial activity, which was evaluated from measurements of soil respiration rate, heat output, and dehydrogenase activity. Significant and year-to-year reproducible differences in all three parameters were observed between soils amended with the three compost types. Some residual (carry-over) effects of previous-year treatments were obtained. Microbial activity decreased in both solarized and non-solarized treatments during the experimental period, probably because of increasing summer temperatures, but the decline was sharper in the solarized treatments probably due to oxygen deficiency under the tarp. Significantly higher activity was found in the treatments amended with the immature compost, both in the solarized and non-solarized treatments. This could be a result of the high level of dissolved organic matter in the immature compost, which enhanced microbial activity.

Microbial communities are highly responsive to environmental cues, and both their structure and activity can be altered in response to changing conditions. We hypothesized that host-associated microbial communities, particularly those colonizing host surfaces, can serve as in situ sensors to reveal environmental conditions experienced by both microorganisms and the host. For a proof-of-concept, we studied a model plant-soil system and employed a non-deterministic gene-centric approach. A holistic analysis was performed using plants of two species and irrigation with water of low quality to induce host stress. Our analyses examined the genetic potential (DNA) and gene expression patterns (RNA) of plant-associated microbial communities, as well as transcriptional profiling of host plants.Transcriptional analysis of plants irrigated with treated wastewater revealed significant enrichment of general stress-associated root transcripts relative to plants irrigated with fresh water. Metagenomic analysis of root-associated microbial communities in treated wastewater-irrigated plants, however, revealed enrichment of more specific stress-associated genes relating to high levels of salt, high pH and lower levels of oxygen. Meta-analysis of these differentially abundant genes obtained from other metagenome studies, provided evidence of the link between environmental factors such as pH and oxygen and these genes. Analysis of microbial transcriptional response demonstrated that enriched gene content was actively expressed, which implies contemporary response to elevated levels of pH and salt.We demonstrate here that microbial profiling can elucidate stress signals that cannot be observed even through interrogation of host transcriptome, leading to an alternate mechanism for evaluating in situ conditions experienced by host organisms. This study is a proof-of-concept for the use of microbial communities as microsensors, with great potential for interrogation of a wide range of host systems. Video Abstract.

Elevated CO2 (eCO2) stimulates plant growth and photosynthesis, which affect root deposition, leading to altered structure and function of the root microbiome. We studied the effect of eCO2 on wheat-root microbiome composition and plant development, with an emphasis on denitrifying communities. Wheat plants were grown in a greenhouse with continuous fertigation for 6 weeks under ambient CO2 (400 ppm) or eCO2 (850 ppm). The total bacterial community was quantified using qPCR with universal 16S rRNA gene primers, and denitrifying genes (nirK, nirS, nosZ) were measured. In addition, total (16S-based) and N2O-reducing (nosZ-based) bacterial community compositions in the soil and roots were analyzed by amplicon sequencing during plant growth. eCO2 had a significant impact on abundance of the studied denitrifying genes, particularly during the late stages of wheat growth before spike formation. Moreover, eCO2 had a significant impact on N2O-reducing community structure in roots. This effect was more pronounced on Burkholderiales and Rhizobiales, with a minor effect on Pseudomonadales. In addition, as expected, bacterial community structure (total and N2O-reducing bacteria), and denitrifying gene abundance, were primary influenced by habitat (soil vs. roots), and secondarily by plant developmental stage. In summary, it is suggested that eCO2 may change root microbiome, enhance wheat development and N demand without an increase in N2O emission.

No-tillage (NT) is a common soil-conservation management practice with known agricultural advantages and drawbacks. However, its short- and long-term effects on the soil microbiome have not been well established. Here, we compared conventional (CT), minimal (MT) and NT practices in two agricultural fields in the north of Israel over a period of 3 years. Edaphic properties, plant-associated pests, weed species abundance and soil microbial community structure were assessed to examine the effects of tillage. Tillage significantly altered physical and chemical soil properties, and a significant increase in hydrolytic and redox microbial activities was observed in NT soils from both sites. Consistent with this, the microbial community structure of NT samples diverged significantly over time from those of CT samples. Repetitive tillage and even a single tillage event caused significant changes in the relative abundance of microorganisms at taxonomic levels ranging from phylum to OTU. However, no significant difference between treatments was found in microbial community alpha-diversity or crop yield. Conversely, higher levels of weed diversity and some pests number were found in NT samples. Overall, we demonstrate that tillage plays a major role in shaping microbial community structure, and in influencing additional environmental, ecological and agricultural soil parameters.

Plant hosts recruit and maintain a distinct root-associated microbiota based on host and bacterium traits. However, past studies disregarded microbial strain-host specificity and spatial micro-heterogeneity of the root compartment. Using genetic manipulation, confocal laser scanning microscopy, real-time quantitative PCR, and genome sequencing we characterized the colonization patterns of three Pseudomonas spp. isolates native to wheat roots, on the micro-scale. Namely, isolates P. fluorescens NT0133, P. stutzeri NT124, and P. stutzeri NT128. All three isolates preferentially colonized wheat over cucumber roots that served as control for host specificity. Furthermore, not only had the isolates strong host specificity but each isolate had a distinct spatial distribution on the root, all within a few millimeters. Isolate P. stutzeri-NT0124 preferentially colonized root tips, whereas P. fluorescens-NT0133 showed a preference for zones distant from the tip. In contrast, isolate P. stutzeri-NT0128 had no preference for a specific niche on the root. While all isolates maintained genetic potential for motility and biofilm formation their phenotype varied significantly and corresponded to their niche preference. These results demonstrate the importance of spatial colonization patterns, governed by both niche and bacterial characteristics which will have great importance in future attempts to manipulate the plant microbiome by constructing synthetic microbial consortia.

Plant roots create specific microbial habitat in the soil - the rhizosphere. In this study, we characterized the rhizosphere microbiome of four host plant species to get insight into the impact of the host (host signature effect) on resident vs. active communities. Results show a distinct plant host specific signature found among wheat, maize, tomato and cucumber, based on the following three parameters: (i) each plant promoted the activity of a unique suite of soil bacterial populations; (ii) significant variations were observed in the number and the degree of dominance of active populations; and (iii) the level of contribution of active (rRNA-based) populations to the resident (DNA-based) community profiles. In the rhizoplane of all four plants, a significant reduction of diversity was observed, relative to the bulk soil. Moreover, an increase in DNA-RNA correspondence indicated higher representation of active bacterial populations in the residing rhizoplane community. This study demonstrates that the host plant determines the bacterial community composition in its immediate vicinity, especially with respect to the active populations.

Summary Limited freshwater (FW) availability due to climate change and increasing global populations is driving agriculture in arid and semi-arid regions to recycle vast quantities of water for irrigation. However, irrigation with treated wastewater (TWW), which contains dissolved organic matter, salts and microorganisms might alter soil microbial populations, and thus affect soil fertility. We characterized the effects of irrigation with TWW and FW on soil bacterial community composition for three consecutive years. Orchard samples were collected at the end of each of the rainy (winter) and irrigation (summer) season. Community composition, determined by 16S ribosomal DNA amplicon pyrosequencing, was highly similar in soil samples obtained at the end of three consecutive rainy seasons, regardless of irrigation season water quality. However, whereas composition in soil shifted slightly during irrigation seasons by FW irrigation, it was greatly influenced by TWW irrigation. During the irrigation season, a decrease in the relative abundance of Actinobacteria was observed; along with an increase in the relative abundance Gammaproteobacteria within TWW-irrigated soils. The return to the ‘baseline state’ during the rainy season demonstrates that the soil community is not resistant to anthropogenic impact imposed by irrigation water quality, yet is resilient in long term.

The formation of biofilms and biofouling is a common feature in aquatic environments. The aim of this study was to identify the primary colonizers of biofilm formed in Eastern Mediterranean Coastal water at different seasons and follow early dynamics of biofilm community development. Pre-treated coastal seawater and biofilm samples were collected from six different sampling events of 2 weeks' duration each during 1 year. The microbial community composition and specific abundance were estimated by 16S rRNA gene clone libraries and fluorescence in situ hybridization-confocal laser scanning microscopy (FISH-CLSM), respectively. The biofilm formed over the course of the year was fairly consistent in terms of community composition and overall abundance with the exception of spring season. Alphaproteobacteria (30-70% of total bacteria), in particular Rhodobacteraceae, were the dominant bacteria in the biofilm, regardless of season, followed by Bacteroidetes (5-35%) and Gammaproteobacteria (6-35%). There was a decrease in relative abundance of Alphaproteobacteria and an increase in the abundance of Bacteroidetes between the initial and 2-week-old biofilm. This observation may aid man-made facilities that have to deal with biofilm formation and help the development of appropriate strategies to control those biofilms.

Colletotrichum and its teleomorph Glomerella are considered major fungal plant pathogens worldwide. They cause significant economic damage to fruit crops in tropical, subtropical and temperate regions. Several Colletotrichum species or biotypes are known to cause disease in a single host such as C. acutatum and C. gloeosporioides on apple, avocado, mango, papaya, passiflora, strawberry and tamarillo. It is also common to find a single species infecting multiple hosts such as C. gloeosporioides on apple, avocado, mango, papaya, peach, strawberry, and other hosts. Cross-infection potential was shown for two species, C. gloeosporioides from limonium and C. acutatum from strawberry, when inoculating peach, pear, mango, nectarine and strawberry. Molecular analyses including species-specific PCR amplification and ITS sequencing was reliable for identification of Colletotrichum isolates infecting apple, avocado, mango, papaya, passiflora, peach, strawberry and tamarillo fruits. Sub-populations within C. acutatum, C. gloeosporioides and C. boninense were characterized using ITS sequence analysis, while the latter appears to be a diverse and emerging new species infecting multiple fruit crops.

In recent years the behavior and properties of nanoparticles released to the environment have been studied extensively to better assess the potential consequences of their broad use in commercial products. The fate, transport and mobility of nanoparticles in soil were shown to be strongly dependent on environmental conditions. However, little is known about the possible effects of nanoparticles on soil chemical, physical and biological properties. In this study, two types of metal oxide nanoparticles, CuO and Fe3O4 were mixed into two types of soil and the effects of the nanoparticles on various soil properties were assessed. Metal oxide nanoparticles were shown previously to catalyze the oxidation of organic pollutants in aqueous suspensions, and they were therefore expected to induce changes in the organic material in the soil, especially upon addition of an oxidant. It was found that the nanoparticles did not change the total amount of organic materials in the soil or the total organic carbon in the soil extract; however, three-dimensional fluorescence spectroscopy demonstrated changes in humic substances. The nanoparticles also affected the soil bacterial community composition, based on denaturing gradient gel electrophoresis (DGGE) fingerprinting, but had little impact on the macroscopic properties of the soil.

The genus Aeromonas is ubiquitous in aquatic environments encompassing a broad range of fish and human pathogens. Aeromonas strains are known for their enhanced capacity to acquire and exchange antibiotic resistance genes and therefore, are frequently targeted as indicator bacteria for monitoring antimicrobial resistance in aquatic environments. This study evaluated temporal trends in Aeromonas diversity and antibiotic resistance in two adjacent semi-intensive aquaculture facilities to ascertain effects of antibiotic treatment on antimicrobial resistance. In the first facility, sulfadiazine-trimethoprim was added prophylactically upon fingerling stocking and water column-associated Aeromonas were monitored periodically over an eleven-month fish-fattening cycle to assess temporal dynamics in taxonomy and antibiotic resistance. In the second facility, Aeromonas were isolated from fish skin ulcers sampled over a three-year period and from pond water samples to assess associations between pathogenic strains to those in the water column. A total of 1200 Aeromonas spp. were isolated, initially screened for sulfadiazine resistance and further screened against five additional antibiotics. In both facilities, strong correlations were observed between sulfadiazine resistance and trimethoprim and tetracycline resistances, whereas correlations between sulfadiazine resistance and ceftriaxone, gentamycin and chloramphenicol resistances were low. Abundance of multi-drug resistant strains as well as sul1, tetA and intI1 gene-harboring strains was significantly higher in profiles sampled during the fish cycle than those isolated prior to stocking and these genes were extremely abundant in the pathogenic strains. Five phylogenetically-distinct Aeromonas clusters were revealed using partial rpoD gene sequence analysis. Interestingly, prior to fingerling stocking the diversity of water column strains was high, and representatives from all five clusters were identified, including an A. salmonicida cluster that harbored all characterized fish skin ulcer samples. Subsequent to stocking diversity was much lower and most water column isolates in both facilities segregated into an A. veronii-associated cluster. This study demonstrated a strong correlation between aquaculture, Aeromonas diversity and antibiotic resistance. It provides strong evidence for linkage between prophylactic and systemic use of antibiotics in aquaculture and the propagation of antibiotic resistance.

Russeting of smooth-skinned potato varieties affect the protective characteristics of the skin against water loss and pathogens invasion. A distinctive russeting disorder, differs from previously reported tuber blemishes, has been observed in the Negev area of Israel and was named ‘Magen russeting’ after one of the repeatedly affected plots. The russeted skin appears as protruding dark-brown patches on the surface of the tubers. Histological studies identified the russet as old skin cells that remain adhered to the newly formed skin layers below them. Phytopathological tests and rDNA pyrosequencing analysis for whole bacterial community suggested that ‘Magen russeting’ is a physiological disorder. Mineral analysis of isolated pure skin samples indicated that russeted skin areas have higher concentrations of Ca and lower concentrations of K than the smooth skin of normal tubers, irrespective of CaCl2 or KCl fertilization of the soil prior to sowing, or of the irrigation regime during the growing season. Nevertheless, application of CaCl2 prior to tuber-seed sowing reduces the rate of affected mature tubers and the severity of the russeting.

Summary Detailed analysis revealed fundamental differences between bacterial association with cucumber (Cucumis sativus) seeds and seedlings roots. Seed colonization by bacteria seems to result from passive encounter between bacteria, conveyed by imbibed soil solution, and the germinating seed. In accordance, the seed-associated bacterial community composition directly reflected that of the germination medium and was characterized by low dominance. Transition from seed to root was marked by a shift in bacterial community composition and in an increase in dominance values. Furthermore, settlement of bacteria on roots was tightly controlled by the specific properties of each root segment. Size and richness of the seed-associated bacterial community were clearly determinate by the community in the germination medium. In contrast, for fully developed and active roots, the medium effect on these parameters was negligible. Perturbation of the seed environment by a pathogen (Pythium aphanidermatum) had major consequences on the seed bacterial community. However, those were mostly related to direct pathogen–bacteria rather than seed–bacteria interactions. In conclusion, simple, even passive processes may determine the initial stage of plant–microbe association during seed germination, prior to extension of the primary root. Therefore, seed germination is a unique phase in the plant life cycle, with respect to its interaction with the below-ground microbiome.

The use of biocides, particularly chlorine, in reverse osmosis (RO) desalination is widely practiced despite documented evidence that although biocides may be advantageous in controlling microbial counts in the water, in some cases they can actually exacerbate biofouling of the membranes. The adverse effects, associated with widespread biocide use, have spurred the need for finding alternative RO pre-treatment disinfection methods. Ultraviolet (UV) disinfection, and specifically medium pressure ultraviolet (MP-UV) disinfection has been considered a possible alternative and is now gaining recognition as a viable disinfection method applicable to RO desalination; however documentation on the effects of UV pre-treatment on RO membrane biofouling is scarce. This paper reports the findings from a four month field study conducted at a brackish water reverse osmosis (BWRO) desalination plant, treating groundwater, in the north of Israel, in which MP-UV was applied as a pre-treatment disinfection step prior to RO...

Amendment of organic wastes and composts to soil is a common agricultural practice. The soil microbiota is responsible for the recycling and mineralization of these organic amendments and their activity affects soil structure and agricultural viability. In this study we assessed the short-term effects of compost amendment on bacterial community composition and activity in agricultural soil. Changes in biomass, respiration and enzymatic activity were correlated with bacterial community profiles, as determined by Denaturing Gradient Gel Electrophoresis (DGGE) of PCR-amplified 16S ribosomal RNA (rRNA) gene fragments and sequencing of relevant bands. Prior to wetting, the community profile of the compost amended soils was almost identical to that of the original compost, implying that the compost bacterial content was much higher than that of the soil. Nonetheless, wetting of compost-amended soils caused an immediate shift in the bacterial community profile resulting in a pattern similar to that of the original soil profile. Initially, in the first three days following wetting, the bacterial community profiles of the control and compost-amended soils were highly similar to each other. However, from day five to day 13, significant differences in the bacterial profiles developed between the control and compost-amended soils, indicating that the compost-related effect on the bacterial community is somewhat delayed. Compost-amended soils showed enhanced rates of nitrification, respiration, hydrolytic and metabolic activities relative to the control soils without compost. They were also characterized by proliferation of communities associated with the Bacteroidetes phylum, which may be explained by the involvement of this clade in the degradation of compost-derived complex organic substrates in the soil microcosms.

Preplant soil fumigation with metam sodium is used worldwide to control soilborne diseases. The development of accelerated degradation of pesticides in soil, including metam sodium, results in reduced pesticide efficacy. Therefore, we studied microbial involvement in accelerated degradation of methyl isothiocyanate (MITC) following repeated soil applications of the parent compound, metam sodium. MITC degradation was reduced in soil with a history of metam sodium applications following sterilization, indicating the key role of microorganisms in accelerated degradation. Accelerated degradation of MITC was induced by inoculation of soil with no previous application of metam sodium with soil with a history of metam sodium applications. We developed a method to extract the active microbial fraction responsible for MITC degradation from soil with a history of metam sodium applications. This concentrated soil extract induced accelerated degradation of MITC when added to two different soils with no previous application of metam sodium. An extensive shift in total bacterial community composition in concentrated soil extracts occurred after a single metam sodium application. Two Oxalobacteraceae strains, MDB3 and MDB10, isolated from Rehovot soil following triple application of metam sodium rapidly degraded MITC in soil with no previous application of metam sodium. Polymerase chain reaction–denaturing gradient gel electrophoresis analysis of bacterial community composition showed relative enrichment of MDB3 following metam sodium application, suggesting its potential in situ involvement in accelerated degradation development in Rehovot soil. Responses of resident Oxalobacteraceae community members to metam sodium applications differed between Rehovot and En Tamar soils. Isolate MDB10 did not induce accelerated degradation of MITC in En Tamar soil and, with the slow dissipation of MITC, soil suppressiveness of accelerated degradation is suggested. The isolation and identification of MITC-degrading bacteria might be helpful in developing tools for managing accelerated degradation.

The idea of using constructed wetlands for the treatment and improving of wastewater emerged in the second half of the last century. Despite relatively wide use of this environmentally friendly technology, relatively little is known about the microbial populations involved in biotransformation and removal of contaminants in this system. The aim of the current study was to investigate the assembly and function of microbial populations in vertical-flow constructed wetland microcosms designed to improve the quality of wastewater after activated sludge treatment. Also, the performance of 3-year-old wetland ponds was investigated. Even though the quality of the influent water was relatively high, improvement in water parameters such as coliform level, ammonia concentration, BOD, and TSS was observed. The performance of the wetland ponds was comparable to that of the microcosms. The microbial community composition of the biofilm formed on the surface of gravel particles in vegetated and plant-free microcosms was studied by denaturing gradient gel electrophoresis (DGGE) and sequencing of 16S rRNA gene fragments. Highly complex bacterial diversity was observed in the biofilm. Cluster analysis of DGGE patterns demonstrated that depth within the wetland microcosm has a stronger effect on microbial community composition of the biofilm formed on wetland matrix than vegetation. Measurements of fluorescein diacetate hydrolysis activity and nitrification potential revealed that hydrolytic activity was affected by both microcosm depth and vegetation presence, whereas nitrification potential was mostly influenced by depth. Resolving the bacterial assemblage of wetland biofilm, which often is considered a black box, will help to understand the interactions involved in the development of diverse and mature biofilm and its function.