Djouher Debiane

Influence of fly ash aided phytostabilisation of Pb, Cd and Zn highly contaminated soils on Lolium perenne and Trifolium repens metal transfer and physiological stress

Due to anthropogenic activities, large extends of soils are highly contaminated by Metal Trace Element (MTE). Aided phytostabilisation aims to establish a vegetation cover in order to promote in situ immobilisation of trace elements by combining the use of metal-tolerant plants and inexpensive mineral or organic soil amendments. Eight years after Coal Fly Ash (CFA) soil amendment, MTE bioavailability and uptake by two plants, Lolium perenne and Trifolium repens, were evaluated, as some biological markers reflecting physiological stress. Results showed that the two plant species under study were suitable to reduce the mobility and the availability of these elements. Moreover, the plant growth was better on CFA amended MTE-contaminated soils, and the plant sensitivity to MTE-induced physiological stress, as studied through photosynthetic pigment contents and oxidative damage was lower or similar. In conclusion, these results supported the usefulness of aided phytostabilisation of MTE-highly contaminated soils.

Lipid content disturbance in the arbuscular mycorrhizal, Glomus irregulare grown in monoxenic conditions under PAHs pollution

Most polycyclic aromatic hydrocarbons (PAHs) are ubiquitous natural and/or anthropogenic pollutants that have adverse effects on the human health and the environment. Little is known about their potential effects on arbuscular mycorrhizal fungi (AMF). Thus, using monoxenic cultures, this work aims to study the impact of increasing concentrations (140 and 280 μM) of two PAHs [anthracene and benzo[a]pyrene (B[a]P)] on Glomus irregulare lipid content in relation with its development. Changes in the total lipids [fatty acids (FA), sterols, phospholipids (PL) and their associated FA (PLFA)] compositions and contents as well as [malondialdehyde (MDA)] production, of the AMF G. irregulare were examined. Direct toxic effects of both PAHs on the AMF were shown as compared to the control culture. The extraradical hyphae length and spore production were drastically restricted in the presence of PAHs. Significant decreases of the main membrane constituents, phosphatidylcholine (PC) and sterols (in particular 24-methycholesterol) were shown in G. irregulare grown under PAHs treatment. Moreover, PAHs exposure caused an oxidative stress in the AMF extraradical structures pointed out by an increase of the lipid peroxidation biomarker production (MDA). All the observed changes were less marked in presence of anthracene, which was found to be less toxic than B[a]P. Taken together, our results suggested that the drastic decrease of the AMF growth under PAHs pollution could partially be explained by depletions in sterols, PC and MDA accumulation.

Propiconazole toxicity on the non-target organism, the arbuscular mycorrhizal fungus, Glomus irregulare.

Arbuscular mycorrhizal fungi (AMF) are obligate symbionts that colonize the roots of most terrestrial plants. Indeed, 80% of vegetal species realize this symbiosis (Bonfante and Perotto, 1995). Plants generally benefit from this AMF association through increased plant nutrient uptake, plant growth and survival rates (Smith and Read, 2008). The symbiotic association may also increase host plant resistance/tolerance against biotic (Hol and Cook, 2005; Akhtar and Siddiqui, 2008) and abiotic stresses, including salinity, drought and pollution (Gerdemann, 1968; Franco-Ramirez et al., 2007; Giri et al., 2007; Sudova et al., 2007; Cartmill et al., 2008; Debiane et al., 2008, 2009; Campagnac et al., 2010). The functioning of AMF may be impaired by cultural practices such as fungicides application (Sukarno et al., 2006). Unfortunately, the use of fungicides is generalized in modern agriculture for the control of fungal diseases. Most of fungicides act directly on essential fungal functions such as respiration, lipid synthesis or cell division (Leroux, 2003). Consequently, they can exhibit undesirable effects on non-target organisms. Among the fungicides, the Sterol Biosynthesis Inhibitor (SBI) family is one of the most used in agriculture (Hewitt, 1998). Four main classes can be distinguished according to their action target site: (i) squalene epoxidation (e.g. naftifine, terbinafine, tolnaftate), (ii) Δ14 demethylation or DMIs (e.g. imazalil, prochloraz, triadimenol, propiconazole), (iii) Δ14reduction and/or Δ8→ Δ7 isomerisation (e.g. fenpropidine, fenpropimorph, tridemorph), (iiii) C4 demethylation (e.g. fenhexamid) (Leroux, 2003). Several studies carried out on SBI fungicide impact on mycorrhizal plants showed contradictory results on the plant growth, on AM fungal development and on the symbiosis functioning (Dodd and Jeffries, 1989; Von Alten et al., 1993; Schweiger and Jacobsen, 1998; Kjoller and Rosendahl, 2000; Schweiger et al., 2001). The use of different experimental procedure in the reported studies (plant species, culture conditions, fungicide formulation, application methods...) did not allow easy comparison with the results obtained and led to some difficulties to give clear conclusion concerning the SBI fungicides effect on AMF (Sancholle et al., 2001).

Benzo[a]pyrene induced lipid changes in the monoxenic arbuscular mycorrhizal chicory roots

Arbuscular mycorrhizal (AM) colonization may be one of the means that protects plants and allows them to thrive on polycyclic aromatic hydrocarbon-polluted soils including the carcinogenic benzo(a)pyrene (B[a]P). To understand the mechanisms involved in the AM symbiosis tolerance to B[a]P toxicity, the purpose of this study was to compare the lipid compositions as well as the contents between mycorrhizal and non-mycorrhizal chicory root cultures grown in vitro under B[a]P pollution. Firstly, B[a]P induced significant decreases of the Glomalean lipid markers: C16:1ω5 and 24-methyl/methylene sterol amounts in AM roots indicating a reduced AM fungal development inside the roots. Secondly, whereas increases in fatty acid amounts after B[a]P application were measured in non-mycorrhizal roots, no changes were shown in mycorrhizal roots. On the other hand, while, after treatment with B[a]P, the total phospholipid contents were unmodified in non-mycorrhizal roots in comparison with the control, drastic reductions were observed in mycorrhizal roots, mainly owing to decreases in phosphatidylethanolamine and phosphatidylcholine. Moreover, B[a]P affected AM root sterols by reducing stigmasterol. In conclusion, the findings presented in this paper have highlighted, for the first time, significant changes in the AM root lipid metabolism under B[a]P pollution and have culminated on their role in the defense/protection mechanisms.

Propiconazole inhibits the sterol 14α-demethylase in Glomus irregulare like in phytopathogenic fungi

The increasing concentrations impact (0.02, 0.2 and 2 mg L(-1)) of a Sterol Biosynthesis Inhibitor (SBI) fungicide, propiconazole, was evaluated on development and sterol metabolism of two non-target organisms: mycorrhizal or non-mycorrhizal transformed chicory roots and the arbuscular mycorrhizal fungus (AMF) Glomus irregulare using monoxenic cultures. In this work, we provide the first evidence of a direct impact of propiconazole on the AMF by disturbing its sterol metabolism. A significant decrease in end-products sterols contents (24-methylcholesterol and in 24-ethylcholesterol) was observed concomitantly to a 24-methylenedihydrolanosterol accumulation indicating the inhibition of a key enzyme in sterol biosynthesis pathway, the sterol 14α-demethylase like in phytopathogenic fungi. A decrease in end-product sterol contents in propiconazole-treated roots was also observed suggesting a slowing down of the sterol metabolism in plant. Taken together, our findings suggest that the inhibition of the both AM symbiotic partners development by propiconazole results from their sterol metabolism alterations.

Calcareous impact on arbuscular mycorrhizal fungus development and on lipid peroxidation in monoxenic roots

The present work underlined the negative effects of increasing CaCO(3) concentrations (5, 10 and 20 mM) both on the chicory root growth and the arbuscular mycorrhizal fungus (AMF) Glomus irregulare development in monoxenic system. CaCO(3) was found to reduce drastically the main stages of G. irregulare life cycle (spore germination, germinative hyphae elongation, root colonization, extraradical hyphae development and sporulation) but not to inhibit it completely. The root colonization drop was confirmed by the decrease in the arbuscular mycorrhizal fungal marker C16:1ω5 amounts in the mycorrhizal chicory roots grown in the presence of CaCO(3). Oxidative damage evaluated by lipid peroxidation increase measured by (i) malondialdehyde (MDA) production and (ii) the antioxidant enzyme peroxidase (POD) activities, was highlighted in chicory roots grown in the presence of CaCO(3). However, MDA formation was significantly higher in non-mycorrhizal roots as compared to mycorrhizal ones. This study pointed out the ability of arbuscular mycorrhizal symbiosis to enhance plant tolerance to high levels of CaCO(3) by preventing lipid peroxidation and so less cell membrane damage.

The arbuscular mycorrhizal Rhizophagus irregularis activates storage lipid biosynthesis to cope with the benzo[a]pyrene oxidative stress.

The phytoremediation assisted by arbuscular mycorrhizal fungi (AMF) could constitute an ecological and economic method to restore polycyclic aromatic hydrocarbon (PAH) polluted soils. Unfortunately, little is known about the PAH impact on the beneficial symbiotic AMF. Using radiolabelling experiments, our work aims to understand how benzo[a]pyrene (B[a]P), a representative of high molecular weight PAH, acts on the AMF lipid metabolism. Our results showed decreases in the sterol precursors as well as in total phospholipid quantities, in link with the [1-(14)C]acetate incorporation decreases in these lipids. Interestingly, a concomitant increase of [1-(14)C]acetate incorporation by 29.5% into phosphatidylcholine with its content decrease in Rhizophagus irregularis extraradical mycelium was observed, suggesting a membrane regeneration. A second concomitant increase (estimated to 69%) of [1-(14)C]acetate incorporation into triacylglycerols (TAG) with the content decrease was also observed. This suggests a fungal TAG biosynthesis activation probably to offset the decrease in storage lipid content when the fungus was grown under B[a]P pollution. In addition, our findings showed that lipase activity was induced by more than 3 fold in the presence of B[a]P in comparison to the control indicating that the drop in TAG content could be a consequence of their active degradation. Taken together, our data suggest the involvement of the fungal TAG metabolism to cope B[a]P toxicity through two means: (i) by providing carbon skeletons and energy necessary for membrane regeneration and/or for B[a]P translocation and degradation as well as (ii) by activating the phosphatidic acid and hexose metabolisms which may be involved in cellular stress defence.