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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.

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.

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.

In this study we used Pisum sativum as model plant, to perform a battery of plant biotests, based on the analysis of biological endpoints, ranging from the macroscopical to the microscopical level, in order to evaluate the toxicity of soils sampled from three different polluted areas (two industrial and one exposed to heavy vehicular traffic). In addition to the conventional germination tests and early root growth analyses, the mitotic index and the percentages of mitotic phases and of aberrations in the root apices were calculated. Moreover, DNA loss and damage were evaluated by flow cytometry and COMET assay, respectively. Root samples from polluted soils showed lower mitotic indices and a higher mitotic aberration percentage and DNA loss in comparison to the controls. Data obtained by COMET tests highlighted the soil genotoxicity, especially in the two industrial areas. All together, our results showed that the three studied sites were characterised by different levels of toxicity: in particula...

BACKGROUND Pest control in post-harvest food storage represents a great challenge in the sustainable prevention of food losses, and insecticide-treated netting may represent a valid alternative to traditional practices such as direct application of insecticides. In our work, the efficacy of a permethrin-incorporated net, in combination with polypropylene or jute sacking, was tested, for the control of S. oryzae. Contamination of maize grain by permethrin from the treated netting was also evaluated. RESULTS A 98% control of S. oryzae was achieved using permethrin-treated netting. Both jute and polypropylene acted as additional barriers, thus increasing efficacy to 100%. However results also showed the contamination of maize kernel by permethrin released from the treated netting. The concentration of residues in maize kernels increased with increasing temperature; however the use of jute or polypropylene significantly reduced (by 87% to 97%) the concentration of residues transferred to maize kernels. CONCLUSIONS Permethrin treated netting provided a high level of efficacy in the post-harvest protection of maize. Several factors influenced permethrin residue concentrations in grains when treated nets are used. Therefore, solutions should be found to prevent contamination of food stored in the bags that are treated with insecticides.

Viticulture is one of the most important productive sectors of agriculture. The vineyards can be subject to a large production of losses, mainly due to vegetative stress or diseases. Esca disease (ED) is one of the most widespread diseases in the vineyards and it is difficult to be predicted. The main symptoms appear at level of wood and leave starting to the end of showing yellow spots which turn to red—brown before their complete drying. This research aims to develop and validate a fast and reliable system able to recognize diseased plants in the vineyard through remote sensing applications. This experiment was carried out in a vineyard in the North–West of Italy on rows of V. vinifera plants cv. Dolcetto and Barbera using a multirotor-UAV (Unmanned Aerial Vehicle) equipped with a multispectral camera. A ground survey was performed in order to assess the plant’s health status and georeference their position. During the field test, the UAV flew at three different flying altitude: 30–50–70 m. The collected images were elaborated using Pix4Dmapper software in order to: (i) extract the vineyard rows, (ii) identify their surface covered by a plant, and (ii) calculate a NDVI map for each plant.

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.

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.

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.

Pest control in post-harvest food storage represents a great challenge in the sustainable prevention of food losses, and insecticide-treated netting may represent a valid alternative to traditional practices such as the direct application of insecticides. In our work, the efficacy of a permethrin-incorporated net, in combination with polypropylene or jute sacking, was tested for the control of Sitophilus oryzae. Contamination of maize grain by permethrin from the treated netting was also evaluated.A 98% control of S. oryzae was achieved using permethrin-treated netting. Both jute and polypropylene acted as additional barriers, increasing efficacy to 100%. The results also showed the contamination of maize kernel by permethrin released from the treated netting. The concentration of residues in maize kernels increased with increasing temperature; however, use of jute or polypropylene significantly reduced (by 87% to 97%) the concentration of residues transferred to maize kernels.Permethrin-treated netting provided a high level of efficacy in the post-harvest protection of maize. Several factors influenced permethrin residue concentrations in grains when treated nets are used. Therefore, solutions should be found to prevent contamination of food stored in the bags that are treated with insecticides. © 2017 Society of Chemical Industry.

Anthocyanins are a group of common phenolic compounds in plants. They are mainly detected in flowers and fruits, are believed to play different important roles such as in the attraction of animals and seed dispersal, and also in the increase of the antioxidant response in tissues directly or indirectly affected by biotic or abiotic stress factors. As a major group of secondary metabolites in plants commonly consumed as food, they are of importance in both the food industry and human nutrition. It is known that arbuscular mycorrhizal (AM) fungi can influence the plant secondary metabolic pathways such as the synthesis of essential oils in aromatic plants, of secondary metabolites in roots, and increase flavonoid concentration. Plant Growth-Promoting Bacteria (PGPB) are able to increase plant growth, improving plant nutrition and supporting plant development under natural or stressed conditions. Various studies confirmed that a number of bacterial species living on and inside the root system are beneficial for plant growth, yield and crop quality. In this work it is shown that inoculation with AM fungi and/or with selected and tested Pseudomonas strains, under conditions of reduced fertilization, increases anthocyanin concentration in the fruits of strawberry.

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.

† 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.

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.

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.

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.

There is increasing interest in the quality of crops because of the implications concerning health, economic revenue, and food quality. Here we tested if inoculation with a mixture of arbuscular mycorrhizal fungi (AMF) and/or two strains of plant growth-promoting bacteria (PGPB), in conditions of reduced chemical inputs, affects the quality and yield of strawberry fruits. Fruit quality was measured by concentrations of soluble sugars, various organic acids, and two vitamins (ascorbic and folic acid). Co-inoculation with the AMF and each of the two PGPB resulted in increased flower and fruit production, larger fruit size, and higher concentrations of sugars and ascorbic and folic acid in comparison with fruits of uninoculated plants. These results provide further evidence that rhizospheric microorganisms affect fruit crop quality and show that they do so even under conditions of reduced chemical fertilization and can thus be exploited for sustainable agriculture.

Poplar is a suitable species for phytoremediation, able to tolerate high concentrations of heavy metals (HMs). Arbuscular mycorrhizal fungi (AMF) form symbiotic associations with the roots of most land plants; they improve nutrient uptake and enhance phytoextraction of HMs while alleviating stress in the host plant. This review summarizes previous results from field and greenhouse studies conducted by us and dealing with this topic. In a field trial on a highly Zn- and Cu-contaminated site, differences in plant survival and growth were observed among 168 clones originating from natural populations of Populus alba L. and Populus nigra L. from northern Italy. After two and a half years from planting, the density, activity and metabolic versatility of the culturable fraction of the soil bacteria in the HM-polluted field was higher in the soil close to where larger poplar plants were growing, in spite of comparable HM concentrations recorded in these soils. One well-performing clone of P. alba (AL35), which accumulated a higher concentration of both metals and had high foliar polyamine (PA) levels, was used for further investigation. In a greenhouse study, AL35 cuttings pre-inoculated with AMF (Glomus mosseae or Glomus intraradices) and then transferred to pots containing soil, collected from the HM-polluted site, displayed growth comparable to that of controls grown on unpolluted soil, in spite of higher Cu and Zn accumulation. Such plants also showed an overall up-regulation of metallothionein (MT) and PA biosynthetic genes, together with increased PA levels. A genome-wide transcriptomic (cDNA-AFLP) analysis allowed the identification of a number of genes, mostly belonging to stress-related functional categories of defense and secondary metabolism, that were differentially regulated in mycorrhizal vs. non mycorrhizal plants. A proteomic analysis revealed that, depending on sampling time, changes in protein profiles were differentially affected by AMF and/or HMs. It is concluded that soil-borne microorganisms affect plant performance on HM-polluted soil. In particular, mycorrhizal plants exhibited increased capacity for phytostabilization of HMs, together with improved growth. Their greater stress tolerance may derive from the protective role of PAs, and from the strong modulation in the expression profiles of stress-related genes and proteins.

Arbuscular mycorrhizal (AM) fungi establish a mutualistic symbiosis with the roots of most plant species. While receiving photosynthates, they improve the mineral nutrition of the plant and can also increase its tolerance towards some pollutants, like heavy metals. Although the fungal symbionts exclusively colonize the plant roots, some plant responses can be systemic. Therefore, in this work a clone of Populus alba L., previously selected for its tolerance to copper and zinc, was used to investigate the effects of the symbiosis with the AM fungus Glomus intraradices on the leaf protein expression. Poplar leaf samples were collected from plants maintained in a glasshouse on polluted (copper and zinc contaminated) or unpolluted soil, after four, six and sixteen months of growth. For each harvest, about 450 proteins were reproducibly separated on 2DE maps. At the first harvest the most relevant effect on protein modulation was exerted by the AM fungi, at the second one by the metals, and at the last one by both treatments. This work demonstrates how importantly the time of sampling affects the proteome responses in perennial plants. In addition, it underlines the ability of a proteomic approach, targeted on protein identification, to depict changes in a specific pattern of protein expression, while being still far from elucidating the biological function of each protein.

We performed a field trial to evaluate the response of different poplar clones to heavy metals. We found that poplar plants of the same clone, propagated by cuttings, had a marked variability of survival and growth in different zones of the field that were characterized by very similar physical–chemical prosperities. Since metal uptake and its accumulation by plants can be affected by soil microorganisms, we investigated soil microbial populations that were collected in proximity to the roots of large and small poplar plants. We used microbiological and molecular tools to ascertain whether bacterial strains or species were associated with large, or small poplars, and whether these were different from those present in the bulk (without plants) soil. We found that the culturable fraction of the bacteria differed in the three cases (bulk soil, small or large poplars). While some taxa were always present, two species ( Chryseobacterium soldanellicola and Variovorax paradoxus ) were only found in the soil where poplars (large or small) were growing, independently from the plant size. Bacterial strains of the genus Flavobacterium were prevalent in the soil with large poplar plants. The existence of different microbial populations in the bulk and in the poplar grown soils was confirmed by the DGGE profiles of the bacterial culturable fractions. Cluster analysis of the DGGE profiles highlighted the clear separation of the culturable fraction from the whole microbial community. The isolation and identification of poplar-associated bacterial strains from the culturable fraction of the microbial community provided the basis for further studies aimed at the combined use of plants and soil microorganisms in the remediation of heavy metal polluted soils.

Three ex situ collections of poplar clones from natural populations of Populus alba and P. nigra growing in northern Italy were assessed for their genetic dissimilarity (GD) by means of amplified fragment length polymorphism (AFLP). The high GD evidenced within populations was exploited for screening 168 clones in a field trial on heavy metal-polluted soil. After one growth season, clonal differences in plant survival and growth were observed. On the basis of performance, six clones were singled out, and used to evaluate copper and zinc accumulation in different organs. Clonal differences in metal concentrations were most evident for leaves and stems; one clone of P. alba (AL35) had a distinctly higher concentration of both metals in the roots. Leaf polyamine (putrescine, spermidine, spermine) profiles correlated with tissue metal concentrations, depending on the clone, plant organ and metal. In particular, the high metal-accumulating clone AL35 exhibited a dramatically higher concentration of free and conjugated putrescine. Overall, the results indicate that, given the high GD of Populus even within populations, it is possible to identify genotypes best suited for soil clean-up, and useful also for investigating physiological markers associated with high metal accumulation/tolerance.