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Algeria is the largest country in Africa characterized by semi-arid and arid sites, located in the North, and hypersaline zones in the center and South of the country. Several autochthonous plants are well known as medicinal plants, having in common tolerance to aridity, drought and salinity. In their natural environment, they live with a great amount of microbial species that altogether are indicated as plant microbiota, while the plants are now viewed as a “holobiont”. In this work, the microbiota of the soil associated to the roots of fourteen economically relevant autochthonous plants from Algeria have been characterized by an innovative metagenomic approach with a dual purpose: (i) to deepen the knowledge of the arid and semi-arid environment and (ii) to characterize the composition of bacterial communities associated with indigenous plants with a strong economic/commercial interest, in order to make possible the improvement of their cultivation. The results presented in this work highlighted specific signatures which are mainly determined by climatic zone and soil properties more than by the plant species.

Bacterial endophytes can colonize plant tissues without harming the plant. Instead, they are often able to increase plant growth and tolerance to environmental stresses. In this work, new strains of bacterial endophytes were isolated from three economically important crop plants (sorghum, cucumber and tomato) grown in three different regions in soils with different management. All bacterial strains were identified by 16S rRNA sequencing and characterized for plant beneficial traits. Based on physiological activities, we selected eight strains that were further tested for their antibiotic resistance profile and for the ability to efficiently colonize the interior of sorghum plants. According to the results of the re-inoculation test, five strains were used to inoculate sorghum seeds. Then, plant growth promotion activity was assessed on sorghum plants exposed to salinity stress. Only two bacterial endophytes increased plant biomass, but three of them delayed or reduced plant salinity stress symptoms. These five strains were then characterized for the ability to produce the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which is involved in the increase of stress tolerance. Pseudomonas brassicacearum SVB6R1 was the only strain that was able to produce this enzyme, suggesting that ACC deaminase is not the only physiological trait involved in conferring plant tolerance to salt stress in these bacterial strains.

Vitis vinifera L. is an economically important crop that can be influenced by soil microorganisms, including arbuscular mycorrhizal fungi (AMF), that establish symbiotic associations with its roots. AMF have beneficial effects on grapevine performance improving water use efficiency and replant success. Most grapevine varieties are susceptible to various diseases, and integrated pest management (IPM) is one of the emerging approaches to perform pest control. In the present study, we examined the AMF communities present in the soil associated to the roots of V. vinifera cv. Pinot Noir (comparing them to those present in a soil not affected by grapevine roots), in a vineyard subjected to IPM at two different phenological stages, using 454 Roche sequencing technology. We proposed a new approach to analyze sequencing data. Most of the taxa were included in the family Glomeraceae. In particular, Glomus sp. Rhizophagus sp. and Septoglomus viscosum were present. The family Archeosporaceae was represented only by the genus Archeospora sp. Different AMF communities were found in the two soils and the importance of the phenological stage in regulating AMF biodiversity was assessed.

Salinity and drought are the most important abiotic stresses hampering crop growth and yield. It has been estimated that arid areas cover between 41% and 45% of the total Earth area worldwide. At the same time, the world’s population is going to soon reach 9 billion and the survival of this huge amount of people is dependent on agricultural products. Plants growing in saline/arid soil shows low germination rate, short roots, reduced shoot biomass, and serious impairment of photosynthetic efficiency, thus leading to a substantial loss of crop productivity, resulting in significant economic damage. However, plants should not be considered as single entities, but as a superorganism, or a holobiont, resulting from the intimate interactions occurring between the plant and the associated microbiota. Consequently, it is very complex to define how the plant responds to stress on the basis of the interaction with its associated plant growth-promoting bacteria (PGPB). This review provides an overview of the physiological mechanisms involved in plant survival in arid and saline soils and aims at describing the interactions occurring between plants and its bacteriome in such perturbed environments. The potential of PGPB in supporting plant survival and fitness in these environmental conditions has been discussed.

The concept of symbiosis can be described as a continuum of interactions between organisms ranging from mutualism to parasitism that can also change over time. Arbuscular mycorrhizal fungi (AMF) are among the most important obligate plant symbionts. Once the symbiosis is well established, mycorrhizal plants are more tolerant to biotic or abiotic stresses, so the AMF relationship with the host plant is generally described as mutualistic. However, little is known about AMF effects on the plant during the early stages of root colonization. The aim of this work was to assess the type of interaction (mutualistic or parasitic) between the arbuscular mycorrhizal (AM) fungus Funelliformis mosseae and Solanum lycopersicum cv. Rio Grande plants, at 7, 14, 21, and 28 days after inoculation (DAI), considering that in the adopted experimental design (one plant per pot), the seedling was the only carbon source for fungus development in the absence of common mycorrhizal networks with other plants. At each harvest, mycorrhizal colonization, shoot and root weights, morphometric parameters, and photosynthetic efficiency were evaluated. The presence of the AM fungus in the tomato root system was observed starting from the 14th DAI, and its level increased over time. Few effects of the fungus presence on the considered parameters were observed, and no stress symptoms ever appeared; so, we can state that the fungus behaved as a mutualistic symbiont during the early stages of plant growth. Moreover, a trend towards a positive effect on plant growth was observed at 28 DAI in mycorrhizal plants.

Previous works have suggested that arbuscular mycorrhizal (AM) fungi can enhance the quality of plant products and fruits. Therefore, the aim of the present study was to investigate the effects of AM fungi on Cucumis melo plant growth, yield, fitness and fruit quality. Inoculated and uninoculated plants of two melon cultivars (Baggio and Expò) were grown in field under common agronomic practices. In addition, a greenhouse test was run on cv. Expò in order to assess the effects of AM inoculation on shoot, leaf and flower development in semi-controlled conditions. In comparison to controls, AM plants flowered earlier, produced more flowers and their pulp contained more sugars and carotenoids. The symbiosis also improved seed size and weight, as well as their germination and root elongation in the first stages of growth in field. Concluding, AM inoculation significantly improved fruit quality in field, under commercial production conditions, also leading to the development of seeds with higher germination rate and increasing plant fitness.

Plant Growth Promoting Bacteria (PGPB) and Arbuscular Mycorrhizal Fungi (AMF) can positively affect plant nutrition and growth. Recent studies have also shown that rhizospheric microorganisms can result in improved fruit features. Aim of this work was to evaluate, in an industrial farming, the effects of three selected biostimulants (consisting of a mix of Plant Growth Promoting Bacteria and Arbuscular Mycorrhizal Fungi), employed in conditions of reduced fertilization on yield, fruit quality and nutritional value. Tomato plants were inoculated with AM fungi and Pseudomonas sp. 19Fv1T or P. fluorescens C7, transplanted and grown in open field under conditions of reduced fertilization. The impact of the microorganisms on the fruit yield and nutritional value was assessed by measuring the production, fruit size and concentration of soluble sugars, organic acids, carotenoids and ascorbate. The size and biomass of tomato fruits were affected by the inocula. Sugar concentration was increased by the selected microorganisms. All the mixtures induced an enhancement of malic acid, while double colonization with AMF and PGPB increased β-carotene concentration in fruits if compared to controls. The results of the present study show that inoculation with soil microorganisms can help to drastically reduce the use of chemical fertilization, maintaining and, in some cases, even improving the tomato fruit yield and quality. This can lead to economical, environmental and human health benefits in relation to the increased sustainability.

Soil salinization and desertification due to climate change are the most relevant challenges for the agriculture of the 21st century. Soil compost amendment and plant growth promoting rhizobacteria (PGP-R) are valuable tools to mitigate salinization and desertification impacts on agricultural soils. Selection of novel halo/thermo-tolerant bacteria from the rhizosphere of glicophytes and halophytes, grown on soil compost amended and watered with 150/300 mM NaCl, was the main objective of our study. Beneficial effects on the biomass, well-being and resilience, exerted on the assayed crops (maize, tomato, sunflower and quinoa), were clearly observable when soils were amended with 20% compost despite the very high soil electric conductivity (EC). Soil compost amendment not only was able to increase crop growth and biomass, but also their resilience to the stress caused by very high soil EC (up to 20 dS m−1). Moreover, compost amendment has proved itself a valuable source of highly halo-(4.0 M NaCl)/thermo tolerant rhizobacteria (55 °C), showing typical PGP features. Among the 13 rhizobacterial isolates, molecularly and biochemically characterized, two bacterial strains showed several biochemical PGP features. The use of compost is growing all around the world reducing considerably for farmers soil fertilization costs. In fact, only in Italy its utilization has ensured, in the last years, a saving of 650 million euro for the farmers, without taking into account the environment and human health benefits. Furthermore, the isolation of halo/thermo-tolerant PGPR strains and their use will allow the recovery and cultivation of hundreds of thousands of hectares of saline and arid soils now unproductive, making agriculture more respectful of agro-ecosystems also in view of upcoming climate change.

Wastewaters generated by the pyrolytic process require treatments to reduce the risks of contamination in rivers, lakes, and coastal waters. Utilizing constructed wetlands is one of the possible approaches according to a Circular Economy System. Plant Growth-Promoting Bacteria (PGPB) and Arbuscular Mycorrhizal Fungi (AMF) can improve plant growth and enhance the bioremediation of wastewater. Two experiments were set up: in the first, a pilot mesocosm was designed to evaluate the effects of a consortium of AM fungi and a PGPB strain on Phragmites australis. After 60 days, the highest plant growth was obtained after inoculation with the combination of microorganisms. In the second experiment, a constructed wetland was built to remediate wastewaters from gasification plant. The plants were efficient in scavenging biological oxygen demand (BOD5), chemical oxygen demand (COD), total fat and oils, hydrocarbons, phenols, aldehydes, surfactants, fluorides, sulfites, sulfates, nitrate, and phosphorus. These data suggest that inoculation of P. australis with AMF and PGPB strains significantly improve the depuration process of wastewaters from gasification plants via constructed wetlands.

Phaseolus vulgaris cv. Billo is an important crop, widely cultivated in Western Piedmont (Italy). It can form symbioses both with arbuscular mycorrhizal fungi (AMF) and bacteria belonging to the Rhizobium genus: AMF can improve plant nutrition and fruit/seed quality, whereas R. leguminosarum is a Gram-negative bacterium able to fix atmospheric nitrogen. In this study, 15 Rhizobium strains, isolated either from root nodules or soil, were characterized by molecular and biochemical point of view (P-solubilization, siderophore and IAA production, intrinsic antibiotic resistance) and one of the selected strains was used for field trial. The aims were: i) to select highly efficient symbiotic bacteria adapted to local conditions and ii) to assess bean seed yield and quality, in response to AMF/rhizobia inoculation at reduced fertilization level. At harvest, yield (pod and seed number and weight) and seed quality (starch, protein, fiber and mineral concentrations), root nodulation and mycorrhizal colonization were measured. Inoculation did not influence yield and fiber concentration. On the contrary, the total seed protein amount was higher in the seeds of rhizobium-inoculated plants in combination or without AMF than in uninoculated plants. The starch content in seeds was increased by rhizobia inoculation. Mg, K and Zn seed accumulation was positively affected by AMF, while the Mn concentration was higher in the presence of rhizobia. In conclusion, plant inoculation strongly affected seed characteristics, improving bean quality also in conditions of reduced fertilization.

The rhizosphere is a hotspot of microbial activity where the release of root exudates stimulates bacterial density and diversity. The majority of the bacterial cells in soil are viable, unculturable, but active. Proteomic tools could be useful in gaining information about microbial community activity and to better understand the real interactions between roots and soil. The aim of this work was to characterize the bacterial community associated with Vitis vinifera cv. Pinot Noir roots using a metaproteome approach. Our results confirmed the large potential of proteomics in describing the environmental microbial communities and their activities: in particular, we showed that bacteria belonging to Streptomyces, Bacillus, Bradyrhizobium, Burkholderia and Pseudomonas genera are the most active in protein expression. Concerning the biological activity of these genera in the rhizosphere, we observed the exclusive presence of the phosphorus metabolic process and the regulation of primary metabolic processes. To our knowledge, this is the first study reporting the rhizosphere proteome of V. vinifera, describing the bacterial community structure and activity of an important ecosystem for the Italian landscape, agriculture and economy.

International audience; Arbuscular mycorrhizal fungi (AMF) colonize the roots of most terrestrial plant species, improving plant growth, nutrient uptake and biotic/abiotic stress resistance and tolerance. Similarly, plant growth promoting bacteria (PGPB) enhance plant fitness and production. In this study, three different AMF (Funneliformis mosseae, Septoglomus viscosum, and Rhizophagus irregularis) were used in combination with three different strains of Pseudomonas sp. (19Fv1t, 5Vm1K and Pf4) to inoculate plantlets of Fragaria x ananassa var. Eliana F1. The effects of the different fungus/bacterium combinations were assessed on plant growth parameters, fruit production and quality, including health-promoting compounds. Inoculated and uninoculated plants were maintained in a greenhouse for 4 months and irrigated with a nutrient solution at two different phosphate levels. The number of flowers and fruits were recorded weekly. At harvest, fresh and dry weights of roots and shoots, mycorrhizal colonization and concentration of leaf photosynthetic pigments were measured in each plant. The following fruit parameters were recorded: pH, titratable acids, concentration of organic acids, soluble sugars, ascorbic acids, and anthocyanidins; volatile and elemental composition were also evaluated. Data were statistically analyzed by ANOVA and PCA/PCA-DA. Mycorrhizal colonization was higher in plants inoculated with R. irregularis, followed by F mosseae and S. viscosum. In general, AMF mostly affected the parameters associated with the vegetative portion of the plant, while PGPB were especially relevant for fruit yield and quality. The plant physiological status was differentially affected by inoculations, resulting in enhanced root and shoot biomass. Inoculation with Pf4 bacterial strain increased flower and fruit production per plant and malic acid content in fruits, while decreased the pH value, regardless of the used fungus. Inoculations affected fruit nutritional quality, increasing sugar and anthocyanin concentrations, and modulated pH, malic acid, volatile compounds and elements. In the present study, we show for the first time that strawberry fruit concentration of some elements and/or volatiles can be affected by the presence of specific beneficial soil microorganisms. In addition, our results indicated that it is possible to select the best plant-microorganism combination for field applications, and improving fruit production and quality, also in terms of health promoting properties.

Phytoremediation is a cost-effective and environment friendly in situ technique for the reclamation of heavy metal-polluted soils. The efficacy of this technique, which relies on tolerant plant species, can be improved by the use of chelating agents. A pot experiment was carried out to evaluate the phytoextraction and phytostabilisation capacities of a white poplar (Populus alba L.) clone named AL35 previously selected for its marked tolerance to copper (Cu) and zinc (Zn). Cuttings were grown on agricultural soil highly contaminated with Cu and Zn, in the presence or not (controls) of a chelant mixture (EDTA/EDDS) known to enhance metal bioavailability and, hence, uptake by plant roots, or the not yet investigated synthetic, highly biodegradable polyaspartic acid (PASP). Both chelant treatments improved the phytostabilisation of Cu and Zn in AL35 plants, whilst the phytoextraction capacity was enhanced only in the case of Cu. Considering that the effectiveness of PASP as phytostabilizer was comparable or better than that of EDTA/EDDS, the low cost of its large-scale chemical synthesis and its biodegradability makes it a good candidate for chelant-enhanced metal phytoextraction from soil while avoiding the toxic side-effects previously described for both EDTA and EDDS.

The aim of this work was to assess the effects of plant-beneficial microorganisms (two Pseudomonas strains and a mixed mycorrhizal inoculum, alone or in combination) on the quality of tomato fruits of plants grown in the field and subjected to reduced fertilization. Pseudomonas strain 19Fv1T was newly characterized during this study. The size and quality of the fruits (concentration of sugars, organic acids and vitamin C) were assessed. The microorganisms positively affected the flower and fruit production and the concentrations of sugars and vitamins in the tomato fruits. In particular, the most important effect induced by arbuscular mycorrhizal (AM) fungi was an improvement of citric acid concentration, while bacteria positively modulated sugar production and the sweetness of the tomatoes. The novelty of the present work is the application of soil microorganisms in the field, in a real industrial tomato farm. This approach provided direct information about the application of inocula, allowed the reduction of chemical inputs and positively influenced tomato quality.

SPE IPM INRA UB CT1; International audience; Arbuscular mycorrhizal (AM) fungi are associated with about 80% of land plants. AM fungi provide inorganic nutrients to plants and in return up to 20% of the plant-fixed CO2 is transferred to the fungal symbionts. Since AM fungi are obligate biotrophs, unraveling how sugars are provided to the fungus partner is a key for understanding the functioning of the symbiosis. In this study, we identified two new monosaccharide transporters from Rhizophagus irregularis (RiMST5 and RiMST6) that we characterized as functional high affinity monosaccharide transporters. RiMST6 was characterized as a glucose specific, high affinity H(+) co-transporter. We provide experimental support for a primary role of both RiMST5 and RiMST6 in sugar uptake directly from the soil. The expression patterns of RiMSTs in response to partial light deprivation and to interaction with different host plants were investigated. Expression of genes coding for RiMSTs was transiently enhanced after 48h of shading and was unambiguously dependent on the host plant species. These results cast doubt on the 'fair trade' principle under carbon-limiting conditions. Therefore, in light of these findings, the possible mechanisms involved in the modulation between mutualism and parasitism in plant-AM fungus interactions are discussed. Copyright 2016 Elsevier Masson SAS. All rights reserved.

Soil microorganisms like arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) are known to positively affect plant health, growth, and nutrition. Their use was often suggested in order to reduce the input of chemicals in agriculture. Recently, it has been observed that AMF and PGPR can also affect some quality features of various crops. In this chapter we review the literature concerning the effects of soil microbes, focusing on AMF and PGPR as the most common plant-associated microorganisms, on the quality of crops. Such effects were considered according to commercial and agronomic plant categories. Current limitations of the available information and some possible future developments have been indicated.

Plants growing under zinc excess store it in leaves, the organs in charge with carbon assimilation. Therefore, leaf modifications can help to understand the plant responses to environmental stresses. In this study, we tested the hypothesis that high zinc concentration can affect leaf anatomy and physiology of Populus alba cv. Villafranca, a tree species with interesting perspectives of application in phytoremediation. Plants were supplemented or not with 300 mg zinc per kg of soil and grown for six months. Green and macroscopically symptom-less leaves from poplars of each treatment were analyzed by light and electron microscopy (TEM, SEM-EDS) and by Raman spectroscopy. LHCII and D1–D2 protein expression were considered to evaluate PSII functioning. Zinc was histochemically detected in the leaves. Zinc strongly altered leaf morphology and ultrastructure, and negatively affected PSII activity decreasing D1 and D2, but not LHCII, protein expression. Zinc was mostly detected in the cell walls of the xylem and of the parenchyma cells surrounding the bundles of zinc treated plants, but not in the controls. A significant increase in the number of calcium oxalate crystals was observed in the leaves of zinc treated plants, but the latter metal was not incorporated in crystals, suggesting an increase of free calcium following zinc accumulation in the cell walls.

Arbuscular mycorrhizae (AM) are the most widespread mutualistic symbioses between the roots of most land plants and a phylum of soil fungi. AM are known to influence plant performance by improving mineral nutrition, protecting against pathogens and enhancing resistance or tolerance to biotic and abiotic stresses. The aim of this study was to investigate the frond proteome of the arsenic hyperaccumulator fern Pteris vittata in plants that had been inoculated with one of the two AM fungi (Glomus mosseae or Gigaspora margarita) with and without arsenic treatment. A protective role for AM fungi colonisation in the absence of arsenic was indicated by the down-regulation of oxidative damage-related proteins. Arsenic treatment of mycorrhizal ferns induced the differential expression of 130 leaf proteins with specific responses in G. mosseae- and Gi. margarita-colonised plants. Up-regulation of multiple forms of glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and enolase, primarily in G. mosseae-inoculated plants, suggests a central role for glycolytic enzymes in arsenic metabolism. Moreover, a putative arsenic transporter, PgPOR29, has been identified as an up-regulated protein by arsenic treatment.

† Background and Aims It is increasingly evident that plant tolerance to stress is improved by mycorrhiza. Thus, suitable plant –fungus combinations may also contribute to the success of phytoremediation of heavy metal (HM)-polluted soil. Metallothioneins (MTs) and polyamines (PAs) are implicated in the response to HM stress in several plant species, but whether the response is modulated by arbuscular mycorrhizal fungi (AMF) remains to be clarified. The aim of the present study was to check whether colonization by AMF could modify growth, metal uptake/translocation, and MT and PA gene expression levels in white poplar cuttings grown on HM-contaminated soil, and to compare this with plants grown on non-contaminated soil. † Methods In this greenhouse study, plants of a Populus alba clone were pre-inoculated, or not, with either Glomus mosseae or G. intraradices and then grown in pots containing either soil collected from a multimetal(Cu and Zn) polluted site or non-polluted soil. The expression of MT and PA biosynthetic genes was analysed in leaves using quantitative reverse transcription–PCR. Free and conjugated foliar PA concentrations were determined in parallel. † Results On polluted soil, AMF restored plant biomass despite higher Cu and Zn accumulation in plant organs, especially roots. Inoculation with the AMF caused an overall induction of PaMT1, PaMT2, PaMT3, PaSPDS1, PaSPDS2 and PaADC gene expression, together with increased free and conjugated PA levels, in plants grown on polluted soil, but not in those grown on non-polluted soil. † Conclusions Mycorrhizal plants of P. alba clone AL35 exhibit increased capacity for stabilization of soil HMs, together with improved growth. Their enhanced stress tolerance may derive from the transcriptional upregulation of several stress-related genes, and the protective role of PAs.

Although Rhizoctonia solani is a cosmopolitan soilborne pathogen, the genus includes isolates with different pathogenicity ranging from high virulence to avirulence. The biocontrol strain Pseudomonas fluorescens P190r and the arbuscular mycorrhizal (AM) fungus Glomus mosseae BEG12 were inoculated alone or in combination in tomato plants infested by the mildly virulent pathogen R. solani #235. Plant growth as well as root morphometric and topological parameters were evaluated. The infection of R. solani was significantly reduced by all the combinations of the beneficial microorganisms. Root systems of R. solani‐infected plants were weakly developed but highly branched with a herring‐bone pattern, while those inoculated with the AM fungus, alone or in combination with the bacterial strain, were longer and more developed, and displayed a dichotomous pattern. The interactions among these three microorganisms affected plant growth and root architecture of tomato plants.

Aims: After the determination of the toxic but nonlethal concentration of NaCl for cucumber, we examined the interaction between an ACC (1-aminocyclopropane-1-carboxylate) deaminase producing bacterial strain and an arbuscular mycorrhizal fungus (AMF) and their effects on cucumber growth under salinity. Methods and Results: In the first experiment, cucumber seedlings were exposed to 0·1, 50, 100 or 200 mmol l−1 NaCl, and plant biomass and leaf area were measured. While seeds exposed to 200 mmol l−1 NaCl did not germinate, plant growth and leaf size were reduced by 50 or 100 mmol l−1 salt. The latter salt cancentration caused plant death in 1 month. In the second experiment, seeds were inoculated with the ACC deaminase-producing strain Pseudomonas putida UW4 (AcdS+), its mutant unable to produce the enzyme (AcdS−), or the AMF Gigaspora rosea BEG9, individually or in combination and exposed to 75 mmol l−1 salt. Plant morphometric and root architectural parameters, mycorrhizal and bacterial colonization and the influence of each micro-organism on the photosynthetic efficiency were evaluated. The AcdS+ strain or the AMF, inoculated alone, increased plant growth, affected root architecture and improved photosynthetic activity. Mycorrhizal colonization was inhibited by each bacterial strain. Conclusions: Salinity negatively affects cucumber growth and health, but root colonization by ACC deaminase-producing bacteria or arbuscular mycorrhizal fungi can improve plant tolerance to such stressful condition. Significance and Impact of the Study: Arbuscular mycorrhizal fungus and bacterial ACC deaminase may ameliorate plant growth under stressful conditions. It was previously shown that, under optimal growth conditions, Ps. putida UW4 AcdS+ increases root colonization by Gi. rosea resulting in synergistic effects on cucumber growth. These results suggest that while in optimal conditions ACC deaminase is mainly involved in the bacteria/fungus interactions, while under stressful conditions this enzyme plays a role in plant/bacterium interactions. This finding is relevant from an ecological and an applicative point of view.

The symbiosis between plant roots and arbuscular mycorrhizal (AM) fungi has been shown to affect both the diversity and productivity of agricultural communities. In this study, we characterized the AM fungal communities of Solanum tuberosum L. (potato) roots and of the bulk soil in two nearby areas of northern Italy, in order to verify if land use practices had selected any particular AM fungus with specificity to potato plants. The AM fungal large-subunit (LSU) rRNA genes were subjected to nested PCR, cloning, sequencing, and phylogenetic analyses. One hundred eighty-three LSU rRNA sequences were analyzed, and eight monophyletic ribotypes, belonging to Glomus groups A and B, were identified. AM fungal communities differed between bulk soil and potato roots, as one AM fungal ribotype, corresponding to Glomus intraradices , was much more frequent in potato roots than in soils (accounting for more than 90% of sequences from potato samples and less than 10% of sequences from soil samples). A semiquantitative heminested PCR with specific primers was used to confirm and quantify the AM fungal abundance observed by cloning. Overall results concerning the biodiversity of AM fungal communities in roots and in bulk soils from the two studied areas suggested that potato roots were preferentially colonized by one AM fungal species, G. intraradices .

Bacteria producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase modulate plant ethylene levels. Decreased ethylene levels increase plant tolerance to environmental stresses and promote legume nodulation. On the contrary, the role of ethylene in mycorrhizal symbiosis establishment is still controversial. In this work, the ACC deaminase-producing strain Pseudomonas putida UW4 AcdS+ and its mutant AcdS−, impaired in ACC deaminase synthesis, were inoculated alone or in combination with the AM fungus Gigaspora rosea on cucumber. Mycorrhizal and bacterial colonization as well as plant growth and morphometric parameters were measured. The influence of each microorganism on the photosynthetic efficiency was evaluated on the second and fourth leaf. The strain AcdS+, but not the AcdS− mutant, increased AM colonization and arbuscule abundance. The mycorrhizal fungus, but not the bacterial strains, promoted plant growth. However, the AcdS+ strain, inoculated with G. rosea, induced synergistic effects on plant biomass, total root length and total leaf projected area. Finally, the photosynthetic performance index was increased by the strain UW4 AcdS+ inoculated in combination with G. rosea BEG9. These results suggest a key role of this enzyme in the establishment and development of AM symbiosis.

It was previously shown that arbuscular mycorrhizal fungi (AMF) exert a significant improvement of growth in a tolerant white poplar (Populus alba L.) clone (AL35) grown on Cu- and Zn-polluted soil via foliar alterations in the levels of defence/stress-related transcripts and molecules. However, nothing is known about the epigenetic changes which occur during tolerance acquisition in response to heavy metals (HMs) in the same mycorrhizal vs. non-mycorrhizal poplar plants. In order to analyse the epigenome in leaves of AL35 plants inoculated or not with AMF and grown in a greenhouse on multimetal polluted or unpolluted soil, the Methylation Sensitive Amplification Polymorphism (MSAP) approach was adopted to detect cytosine DNA methylation. Modest changes in cytosine methylation patterns were detected at first sampling (4 months from planting), whereas extensive alterations (hypomethylation) occurred at second sampling (after 6 months) in mycorrhizal plants grown in the presence of HMs. The sequencing of MSAP fragments led to the identification of genes belonging to several Gene Ontology categories. Seven MSAP fragments, selected on the basis of DNA methylation status in treated vs control AL35 leaves at the end of the experiment, were analysed for their transcript levels by means of qRT-PCR. Gene expression varied in treated samples relative to controls in response to HMs and/or AMF inoculation; in particular, transcripts of genes involved in RNA processing, cell wall and amino acid metabolism were upregulated in the presence of AMF with or without HMs.

Aromatic hydrocarbons are pollutants which have mutagenic and carcinogenic properties as well as relatively high hydrosolubility. Their presence in soils makes techniques such as bioremediation an important topic for research. In this work, the effect of arbuscular mycorrhiza (AM) on the persistence of benzene, toluene, ethylbenzene and xylene (BTEX) in artificially contaminated substrates was evaluated. Leek plants were grown with three AM fungal species using a specially designed mesocosm system, in which internal air and substrate samples were analyzed by gas chromatography for BTEX content. Strong reductions in the BTEX concentration in substrates were generally observed in the presence of mycorrhizal plants. Residual BTEX content ranged between nearly total disappearance (

The localization, viability, and culturability of Pseudomonas fluorescens 92 rkG5 were analyzed on three morphological root zones (root tip + elongation, root hair, and collar) of 3-, 5-, and 7-day-old tomato plants. Qualitative information about the localization and viability was collected by confocal laser scanning microscopy. Quantitative data concerning the distribution, viability, and culturability were obtained through combined dilution plating and flow cytometry. Colonization by P. fluorescens affected root development in a complex way, causing a general increase in the length of the collar and early stimulation of the primary root growth (3rd day), followed by a reduction in length (7th day). The three root zones showed different distribution, organization, and viability of the bacterial cells, but the distribution pattern within each zone did not change with time. Root tips were always devoid of bacteria, whereas with increasing distance from the apex, microcolonies or strings of cells became more and more prominent. Viability was high in the elongation zone, but it declined in the older parts of the roots. The so-called viable but not culturable cells were observed on the root, and their proportion in the distal (root tip + elongation) zone dramatically increased with time. These results suggest the existence of a specific temporal and spatial pattern of root colonization, related to cell viability and culturability, expressed by the plant-beneficial strain P. fluorescens 92 rkG5.

I norder to better understand which are the factors involved in the control of nuclear size, and more pre- cisely, to clarify why nuclear hypertrophy doesn't always follow genome increase, biological techniques combined with a strict mathematical approach have been applied to study the nuclei of root cortical cells of Lycopersicon escu- lentum Mill cv Early Mech. Tomato is a multiploid plant, with three different levels of DNA content, therefore it is especially suitable to study nucleus size according to genome size. In addition, as arbuscular mycorrhizal coloniza- tion strongly affects nucleus organization, part of the plants were inoculated with the arbuscular mycorrhizal fungus Glomus mosseae BEG 12. Quantitative and qualitative nuclear changes have been analyzed in mycorrhizal and non- mycorrhizal roots of tomato, by means of microscopy, immuno-labelling and flow cytometry. The results, supported by mathematical analysis, clearly show that increased ploidy is necessary, but not sufficient, to explain nuclear hyper- trophy, in which chromatin decondensation is also involved and related to cell metabolic activity.