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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 HMpolluted 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM PUBLISHER]

Abstract: Significant improvement of growth associated with increased, rather than decreased, uptake of Cu and Zn has been observed in poplar plants inoculated with Glomus spp. as compared with non-mycorrhizal plants. The beneficial effect exerted by these arbuscular mycorrhizal fungi (AMF) is likely to be controlled by specific gene expression patterns in the plant. Until now, however, very little is known about the transcriptional changes which occur in response to heavy metals (HMs) in mycorrhizal vs. non-mycorrhizal poplar plants. In order to identify such HM- and/or AMF-induced changes in leaves of white poplar (Populus alba L.) plants grown, in the greenhouse, on Cu- and Zn-polluted soil, the cDNA-Amplified Fragment Length Polymorphism (AFLP) approach was adopted, resulting in the identification of a number of new differentially regulated genes. Transcript derived fragments (TDFs) mostly belonged to stress-related functional categories of defence and secondary metabolism. Genes belonging to different functional categories, plus other genes known to be related to HM stress (metallothioneins, phytochelatin synthase, glutathione synthase, arginine decarboxylase), were analysed by quantitative (q)RT-PCR. Transcript levels were generally down-regulated, or unaffected, in polluted soil compared with controls, the main exceptions being phytochelatin synthase and clathrin, and strongly up-regulated in the presence of AMF, especially Glomus mosseae. [Copyright &y& Elsevier]

Poplar is a good candidate for phytoremediation purposes because of its rapid growth, extensive root system, and ease of propagation and transformation; however its tolerance to heavy metals has not been fully investigated yet. In the present work, an in vitro model system with shoot cultures was used to investigate the tolerance to high concentrations of zinc (Zn) of a commercial clone (Villafranca) of Populus alba. Based on chlorophyll content (leaf chlorosis) and the rate of adventitious root formation from shoot cuttings as parameters of damage, 0. 5–4mM zinc concentrations were all toxic albeit to different extents. Northern blot and reverse transcriptase (RT)-PCR analyses were used to examine the expression profiles of types 1, 2 and 3 PaMT genes in stems, leaves and roots of plants exposed to Zn treatments. In leaves, MT1 and MT3 mRNA levels were enhanced by Zn, while MT2 transcripts were not affected. The PaMT expression profiles were differentially affected by Zn in an organ-specific manner, and the relationship with Zn concentration and exposure time was rarely linear. The developmental and molecular data reveal that the in vitro model is a sensitive and reliable system to study heavy metal stress responses. [Copyright &y& Elsevier]