Frédéric Laruelle

Sterol distribution in arbuscular mycorrhizal fungi

Abstract The sterol composition of spores from 16 species of arbuscular mycorrhizal fungi belonging to the order Glomales were examined by GC–MS. The major compound was found to be 24-ethylcholesterol (up to 85%) followed by cholesterol (up to 15%). Several other sterols such as 24-methylcholesterol, Δ5-avenasterol and 24-ethylcholesta-5,22-dien-3 β -ol were also detected. Significant amounts of α -amyrin, a common vascular plant triterpene, were present in the spores of all the fungal species analyzed. The absence of ergosterol, a classical fungal sterol, is discussed in relation to fungal evolution.

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.

Differential effects of fenpropimorph and fenhexamid, two sterol biosynthesis inhibitor fungicides, on arbuscular mycorrhizal development and sterol metabolism in carrot roots.

Sterols composition of transformed carrot roots incubated in presence of increasing concentrations of fenpropimorph (0.02; 0.2; 2mgl(-1)) and fenhexamid (0.02; 0.2; 2; 20mgl(-1)), colonized or not by Glomus intraradices was determined. In mycorrhizal roots treated with fenpropimorph, normal Delta(5)-sterols were replaced by unusual compounds such as 9beta,19-cyclopropylsterols (24-methylpollinastanol), Delta(8,14)-sterols (ergosta-8,14-dienol, stigmasta-8,14-dienol), Delta(8)-sterols (Delta(8) sitosterol) and Delta(7)-sterols (ergosta-7,22-dienol). After application of fenpropimorph, a drastic reduction of the mycorrhizal root growth, root colonization and extraradical fungal development was observed. Application of fenhexamid did not modify sterol profiles and the total colonization of roots. But the arbuscule frequency of the fungal partner was significantly affected. Comparison of the effects caused by the tested fungicides indicates that the usual phytosterols may be involved in symbiosis development. Indeed, observed modifications of root sterols composition could explain the high fenpropimorph toxicity to the AM symbiosis. However, the absence of sterolic modifications in the roots treated with fenhexamid could account for its more limited impact on mycorrhization.

Solubility, photostability and antifungal activity of phenylpropanoids encapsulated in cyclodextrins

Effects of the encapsulation in cyclodextrins (CDs) on the solubility, photostability and antifungal activities of some phenylpropanoids (PPs) were investigated. Solubility experiments were carried out to evaluate the effect of CDs on PPs aqueous solubility. Loading capacities and encapsulation efficiencies of freeze-dried inclusion complexes were determined. Moreover, photostability assays for both inclusion complexes in solution and solid state were performed. Finally, two of the most widespread phytopathogenic fungi, Fusarium oxysporum and Botrytis cinerea, were chosen to examine the antifungal activity of free and encapsulated PPs. Results showed that encapsulation in CDs significantly increased the solubility and photostability of studied PPs (by 2 to 17-fold and 2 to 44-fold, respectively). Free PPs revealed remarkable antifungal properties with isoeugenol showing the lowest half-maximal inhibitory concentration (IC50) values of mycelium growth and spore germination inhibition. Encapsulated PPs, despite their reduced antifungal activity, could be helpful to solve drawbacks such as solubility and stability.

Arbuscular mycorrhizal fungal inoculation protects Miscanthus × giganteus against trace element toxicity in a highly metal-contaminated site

Arbuscular mycorrhizal fungus (AMF)-assisted phytoremediation could constitute an ecological and economic method in polluted soil rehabilitation programs. The aim of this work was to characterize the trace element (TE) phytoremediation potential of mycorrhizal Miscanthus × giganteus. To understand the mechanisms involved in arbuscular mycorrhizal symbiosis tolerance to TE toxicity, the fatty acid compositions and several stress oxidative biomarkers were compared in the roots and leaves of Miscanthus × giganteus cultivated under field conditions in either TE-contaminated or control soils. TEs were accumulated in greater amounts in roots, but the leaves were the organ most affected by TE contamination and were characterized by a strong decrease in fatty acid contents. TE-induced oxidative stress in leaves was confirmed by an increase in the lipid peroxidation biomarker malondialdehyde (MDA). TE contamination decreased the GSSG/GSH ratio in the leaves of exposed plants, while peroxidase (PO) and superoxide dismutase (SOD) activities were increased in leaves and in whole plants, respectively. AMF inoculation also increased root colonization in the presence of TE contamination. The mycorrhizal colonization determined a decrease in SOD activity in the whole plant and PO activities in leaves and induced a significant increase in the fatty acid content in leaves and a decrease in MDA formation in whole plants. These results suggested that mycorrhization is able to confer protection against oxidative stress induced by soil pollution. Our findings suggest that mycorrhizal inoculation could be used as a bioaugmentation technique, facilitating Miscanthus cultivation on highly TE-contaminated soil.

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.

Impact of soil salinity on arbuscular mycorrhizal fungi biodiversity and microflora biomass associated with Tamarix articulata Vahll rhizosphere in arid and semi-arid Algerian areas

Soil salinization is an increasingly important problem in many parts of the world, particularly under arid and semi-arid areas. Unfortunately, the knowledge about restoration of salt affected ecosystems using mycorrhizae is limited. The current study aims to investigate the impact of salinity on the microbial richness of the halophytic plant Tamarix articulata rhizosphere. Soil samples were collected from natural sites with increasing salinity (1.82-4.95 ds.m(-1)). Six arbuscular mycorrhizal fungi (AMF) species were isolated from the different saline soils and identified as Septoglomus constrictum, Funneliformis mosseae, Funneliformis geosporum, Funneliformis coronatum, Rhizophagus fasciculatus, and Gigaspora gigantea. The number of AMF spores increased with soil salinity. Total root colonization rate decreased from 65 to 16% but remained possible with soil salinity. Microbial biomass in T. articulata rhizosphere was affected by salinity. The phospholipid fatty acids (PLFA) C16:1ω5 as well as i15:0, a15:0, i16:0, i17:0, a17:0, cy17:0, C18:1ω7 and cy19:0 increased in high saline soils suggesting that AMF and bacterial biomasses increased with salinity. In contrast, ergosterol amount was negatively correlated with soil salinity indicating that ectomycorrhizal and saprotrophic fungal biomasses were reduced with salinity. Our findings highlight the adaptation of arbuscular and bacterial communities to natural soil salinity and thus the potential use of mycorrhizal T. articulata trees as an approach to restore moderately saline disturbed arid lands.

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.