Phytomanagement of a metal-contaminated agricultural soil with Sorghum bicolor, humic / fulvic acids and arbuscular mycorrhizal fungi near the former Pb/Zn metaleurop Nord smelter.

As many contaminated agricultural soils can no longer be used for food crops, lignocellulosic energy crops matter due to their ability to grow on such soils and to produce biomass for biosourced materials and biofuels, thereby reducing the pressure on the limited arable lands. Sorghum bicolor (L.) Moench, can potentially produce a high biomass suitable for producing bioethanol, renewable gasoline, diesel, and sustainable aircraft fuel, despite adverse environmental conditions (e.g. drought, contaminated soils). A 2-year field trial was carried out for the first time in the northern France for assessing sorghum growth on a Cd, Pb and Zn-contaminated agricultural soil amended with humic/fulvic acid, alone and paired with arbuscular mycorrhizal fungi. Sorghum produced on average (in t DW ha-1): 12.4 in year 1 despite experiencing a severe drought season and 15.3 in year 2. Humic/fulvic acids (Lonite 80SP®) and arbuscular mycorrhizal fungi did not significantly act as biostimulants regarding the shoot DW yield and metal uptake of sorghum. The annual shoot Cd, Pb and Zn removals averaged 0.14, 0.20 and 1.97 kg ha-1, respectively. Sorghum cultivation and its metal uptake induced a significant decrease in 0.01 M Ca(NO3)2-extractable soil Cd, Pb and Zn concentrations by 95%, 73% and 95%, respectively, in year 2. Soluble and exchangeable soil Cd, Pb and Zn would be progressively depleted in subsequent crops, which should result in lower pollutant linkages and enhanced ecosystem services. This evidenced sorghum as a relevant plant species for phytomanaging the large area (750 ha) with metal-contaminated soil near the former Pb/Zn Metaleurop Nord smelter, amidst ongoing climate change. The potential bioethanol yield of the harvested sorghum biomass was 5589 L ha-1. Thus sorghum would be a promising candidate for bioethanol production, even in this northern French region.

Plant testing with hemp and miscanthus to assess phytomanagement options including biostimulants and mycorrhizae on a metal-contaminated soil to provide biomass for sustainable biofuel production.

The need of biofuels from biomass, including sustainable aviation fuel, without using agricultural land dedicated to food crops, is in constant demand. Strategies to intensify biomass production using mycorrhizal fungi, biostimulants and their combinations could be solutions for improving the cultivation of lignocellulosic plants but still lack well-established validation on metal-contaminated soils. This study aimed to assess the yield of Miscanthus x giganteus J.M. Greef & Deuter and Cannabis sativa L. grown on a metal-contaminated agricultural soil (11 mg Cd, 536 mg Pb and 955 mg Zn kg-1) amended with biostimulants and/or arbuscular mycorrhizal fungi, and the shoot Cd, Pb and Zn uptake. A pot trial was carried out with soil collected from a field near a former Pb/Zn smelter in France and six treatments: control (C), protein hydrolysate (a mixture of peptides and amino acids, PH), humic/fulvic acids (HFA), arbuscular mycorrhizae fungi (AMF), PH combined with AMF (PHxAMF), and HFA combined with AMF (HFAxAMF). Metal concentrations in the soil pore water (SPW), pH and electrical conductivity were measured over time. Miscanthus and hemp shoots were harvested on day 90. Both PH and PHxAMF treatments increased SPW Cd, Pb, and Zn concentrations (e.g. by 26, 1.9, and 22.9 times for miscanthus and 9.7, 4.7, and 19.3 times for hemp in the PH and PHxAMF treatments as compared to the control one, respectively). This led to phytotoxicity and reduced shoot yield for miscanthus. Conversely, HFA and HFAxAMF treatments decreased SPW Cd and Zn concentrations, increasing shoot yields for hemp and miscanthus. Shoot Cd, Pb, and Zn uptakes peaked for PH and PHxAMF hemp plants (in µg plant-1, Cd: 310-334, Pb: 34-38, and Zn: 232-309 for PHxAMF and PH, respectively), while lowest values occurred for PH miscanthus plants mainly due to low shoot yield. Overall, this study suggested that humic/fulvic acids can be an effective biostimulant for increasing shoot biomass production in a metal-contaminated soil. These results warrant further investigations of the HFAxAMF in field trials.

Coriander (Coriandrum sativum) Cultivation Combined with Arbuscular Mycorrhizal Fungi Inoculation and Steel Slag Application Influences Trace Elements-Polluted Soil Bacterial Functioning.

The cultivation of aromatic plants for the extraction of essential oils has been presented as an innovative and economically viable alternative for the remediation of areas polluted with trace elements (TE). Therefore, this study focuses on the contribution of the cultivation of coriander and the use of arbuscular mycorrhizal fungi (AMF) in combination with mineral amendments (steel slag) on the bacterial function of the rhizosphere, an aspect that is currently poorly understood and studied. The introduction of soil amendments, such as steel slag or mycorrhizal inoculum, had no significant effect on coriander growth. However, steel slag changed the structure of the bacterial community in the rhizosphere without affecting microbial function. In fact, Actinobacteria were significantly less abundant under slag-amended conditions, while the relative proportion of Gemmatimonadota increased. On the other hand, the planting of coriander affects the bacterial community structure and significantly increased the bacterial functional richness of the amended soil. Overall, these results show that planting coriander most affected the structure and functioning of bacterial communities in the TE-polluted soils and reversed the effects of mineral amendments on rhizosphere bacterial communities and their activities. This study highlights the potential of coriander, especially in combination with steel slag, for phytomanagement of TE-polluted soils, by improving soil quality and health.

New plant immunity elicitors from a sugar beet byproduct protect wheat against Zymoseptoria tritici.

The current worldwide context promoting agroecology and green agriculture require the discovery of new ecofriendly and sustainable plant protection tools. Plant resistance inducers, called also elicitors, are one of the most promising alternatives fitting with such requirements. We produced here a set of 30 molecules from pyroglutamic acid, bio-sourced from sugar beet byproducts, and examined for their biological activity on the major agro-economically pathosystem wheat-Zymoseptoria tritici. Foliar application of the molecules provided significant protection rates (up to 63% disease severity reduction) for 16 among them. Structure-activity relationship analysis highlighted the importance of all chemical groups of the pharmacophore in the bioactivity of the molecules. Further investigations using in vitro and in planta antifungal bioassays as well as plant molecular biomarkers revealed that the activity of the molecules did not rely on direct biocide activity towards the pathogen, but rather on the activation of plant defense mechanisms dependent on lipoxygenase, phenylalanine ammonia-lyase, peroxidase, and pathogenesis-related protein pathways. This study reports a new family of bio-sourced resistance inducers and provides new insights into the valorization of agro-resources to develop the sustainable agriculture of tomorrow.

Coriander (Coriandrum sativum L.) in Combination with Organic Amendments and Arbuscular Mycorrhizal Inoculation: An Efficient Option for the Phytomanagement of Trace Elements-Polluted Soils.

The cultivation of coriander (Coriandrum sativum L.) destined for essential oils production was recently presented as an innovative and economically viable alternative for the phytomanagement of trace elements (TE)-polluted soils. However, Cd accumulation in shoots has proven to be an obstacle in the valorization of the distillation residues and the development of these phytotechnologies. The present study aimed to evaluate the effect of arbuscular mycorrhizal fungus (Funneliformis mosseae) inoculation and organic amendment application on the soil TE bioavailability and plant uptake, as well as on the soil quality and health improvement. The application of compost and sewage sludge improved the growth of coriander and Cd and Zn immobilization in soil, resulting in reduced Cd plant uptake. A synergistic effect of arbuscular mycorrhizal fungi (AMF) inoculation and organic amendments was observed in the decrease in the extractable soil Cd and Zn concentrations, but not in the Cd plant uptake. Despite a significant decrease in Cd accumulation in shoots, coriander retained its accumulative phenotype, with a metal bioconcentration factor close to 1. Furthermore, both the vegetation and the organic amendments improved the soil quality and health by increasing its microbial biomass, as estimated by phospholipid fatty acids, soil enzyme activities (dehydrogenase, phosphatase, ß-glucosidase, and cellubiosidase), and the bacterial metabolic function and diversity. The findings demonstrate the potential of C. sativum, particularly in combination with organic amendments and AMF inoculation, for the phytomanagement of TE-polluted soils and soil quality and health improvement.

In situ cultivation of aromatic plant species for the phytomanagement of an aged-trace element polluted soil: Plant biomass improvement options and techno-economic assessment of the essential oil production channel.

Due to the presence of trace element (TE) in agricultural soils, wide areas are unproper for food production and the clean-up of soil is not a feasible option. Considering the potential remediation options, the use of aromatic plants producing a high quantity of biomass and developing high-added value sectors such as essential oil (EO) production could be valuable regarding one of the phytomanagement objectives, i.e. the restoration of an economic activity. The purpose of this study was hence to evaluate in situ the suitability of two aromatic crops, clary sage and coriander, for the phytomanagement of aged TE-polluted soils, taking into account plants' growth, development and biomass production, essential oil (EO) content and quality as well as a techno-economic feasibility analysis of the channel. In situ experiments have been carried out on two agricultural plots of 1.5 ha, a TE-polluted one (Pb: 394 ppm - Zn: 443 ppm - Cd: 7.2 ppm) and an unpolluted one (Pb: 22 ppm - Zn: 48 ppm - Cd: 0.4 ppm). Our findings have shown the ability of coriander and sage to grow similarly on both unpolluted and TE-polluted soil and to produce significant amounts of biomass. The pesticide residue and TE analyses have demonstrated that the EO only contained trace amounts of the contaminants, below or close to the limit of quantification of the method used and similar to marketed products. Mycorrhizal inoculation has also shown promising results by increasing the colonization rates of both aromatic plants, but did not result in higher biomass or EO amounts. Our study brings new evidence towards the potential of clary sage to be used for the phytomanagement of TE-polluted areas, given its perennial vegetation cover, tolerance to TE and obtained EO yields.

Aided Phytoremediation to Clean Up Dioxins/Furans-Aged Contaminated Soil: correlation between microbial communities and pollutant dissipation.

To restore and clean up polluted soils, aided phytoremediation was found to be an effective, eco-friendly, and feasible approach in the case of many organic pollutants. However, little is known about its potential efficiency regarding polychlorinated dibenzo-p-dioxins and furans-contaminated soils. Thus, phytoremediation of aged dioxins/furans-contaminated soil was carried out through microcosm experiments vegetated with alfalfa combined with different amendments: an arbuscular mycorrhizal fungal inoculum (Funneliformis mosseae), a biosurfactant (rhamnolipids), a dioxins/furans degrading-bacterium (Sphingomonas wittichii RW1), and native microbiota. The total dioxins/furans dissipation was estimated to 23%, which corresponds to 48 ng.kg-1 of soil, after six months of culture in the vegetated soil combined with the four amendments compared to the non-vegetated soil. Our findings showed that the dioxins/furans dissipation resulted from the stimulation of soil microbial enzyme activities (fluorescein diacetate hydrolase and dehydrogenase) and the increase of bacterial abundance, richness, and diversity, as well as fungal diversity. Amplicon sequencing using Illumina MiSeq analysis led to identification of several bacterial (Bacillaceae, Sphingomonadaceae) and fungal (Chaetomium) groups known to be involved in dioxins/furans degradation. Furthermore, concomitant cytotoxicity and dioxins/furans concentration decreases were pointed out in the phytoremediated soil. The current study demonstrated the usefulness of combining different types of amendments to improve phytoremediation efficacy of aged dioxins/furans-contaminated soils.

Initial microbial status modulates mycorrhizal inoculation effect on rhizosphere microbial communities.

Arbuscular mycorrhizal fungi (AMF) play a central role in rhizosphere functioning as they interact with both plants and soil microbial communities. The conditions in which AMF modify plant physiology and microbial communities in the rhizosphere are still poorly understood. In the present study, four different plant species, (clover, alfalfa, ryegrass, tall fescue) were cultivated in either sterilized (γ ray) or non-sterilized soil and either inoculated with a commercial AMF (Glomus LPA Val 1.) or not. After 20 weeks of cultivation, the mycorrhizal rate and shoot and root biomasses were measured. The abundance and composition of bacteria, archaea, and fungi were analyzed, respectively, by quantitative PCR (qPCR) and fingerprinting techniques. Whilst sterilization did not change the AMF capacity to modify plant biomass, significant changes in microbial communities were observed, depending on the taxon and the associated plant. AMF inoculation decreases both bacterial and archaeal abundance and diversity, with a greatest extent in sterilized samples. These results also show that AMF exert different selections on soil microbial communities according to the plant species they are associated with. This study suggests that the initial abundance and diversity of rhizosphere microbial communities should be considered when introducing AMF to cultures.

Genetic Structure of Zymoseptoria tritici in Northern France at Region, Field, Plant, and Leaf Layer Scales.

Population genetic structure of the worldwide-distributed wheat pathogen Zymoseptoria tritici has been extensively studied at large geographical scales, but to a much less extent at small or local spatial scales. A total of 627 single-conidial fungal isolates were sampled from several locations in northern France (Hauts-de-France Region) to assess fungal genetic structure at region, field, plant, and leaf layer scales, using highly polymorphic microsatellite markers and mating type idiomorphs. Important and overall similar levels of both gene and genotype diversities (gene diversity values of ≥0.44 and haplotype frequencies of ≥94%) were found at all the examined scales. Such rates of diversity are likely due to an active sexual recombination in the investigated areas, as revealed by equal proportions of the two mating types scored in all sampled populations. Interestingly, a rare occurrence of clones among lesions from a same leaf, as well as among leaves from different plant leaf layers (e.g., upper versus lower leaves), was highlighted, indicating that ascospores contribute much more than expected to Z. tritici epidemics, compared with pycnidiospores. Population structure and analyses of molecular variance revealed significant genetic differentiation at the regional scale (GST = 0.23) and, as expected, not at the other more local scales (GST ≤ 0.01). Further analyses using Bayesian and unweighted neighbor-joining statistical methods detected six genetic clusters within the regional population, overall distributed according to the locations from which the isolates were sampled. Neither clear directional relative migration linked to the geographical distribution of the locations, nor isolation by distance, were observed. Separate evolutionary trajectories caused by selection and adaptations to habitat heterogeneity could be the main forces shaping such structuration. This study provides new insights into the epidemiology and the genetic structure of Z. tritici at small local and, for the first time, at single plant and leaf layer scales. Such findings would be helpful in implementing effective control strategies.

Biocontrol of the wheat pathogen Zymoseptoria tritici using cyclic lipopeptides from Bacillus subtilis.

Innovation toward ecofriendly plant protection products compatible with sustainable agriculture and healthy food is today strongly encouraged. Here, we assessed the biocontrol activity of three cyclic lipopeptides from Bacillus subtilis (mycosubtilin, M; surfactin, S; fengycin, F) and two mixtures (M + S and M + S + F) on wheat against Zymoseptoria tritici, the main pathogen on this crop. Foliar application of these biomolecules at a 100-mg L-1 concentration on the wheat cultivars Dinosor and Alixan, 2 days before fungal inoculation, provided significant reductions of disease severity. The best protection levels were recorded with the M-containing formulations (up to 82% disease reduction with M + S on Dinosor), while S and F treatments resulted in lower but significant disease reductions. In vitro and in planta investigations revealed that M-based formulations inhibit fungal growth, with half-maximal inhibitory concentrations of 1.4 mg L-1 for both M and M + S and 4.5 mg L-1 for M + S + F, thus revealing that the observed efficacy of these products may rely mainly on antifungal property. By contrast, S and F had no direct activity on the pathogen, hence suggesting that these lipopeptides act on wheat against Z. tritici as resistance inducers rather than as biofungicides. This study highlighted the efficacy of several lipopeptides from B. subtilis to biocontrol Z. tritici through likely distinct and biomolecule-dependent modes of action.

Beneficial contribution of the arbuscular mycorrhizal fungus, Rhizophagus irregularis, in the protection of Medicago truncatula roots against benzo[a]pyrene toxicity.

Arbuscular mycorrhizal fungi are able to improve plant establishment in polluted soils but little is known about the genes involved in the plant protection against pollutant toxicity by mycorrhization, in particular in the presence of polycyclic aromatic hydrocarbons (PAH). The present work aims at studying in both symbiotic partners, Medicago truncatula and Rhizophagus irregularis: (i) expression of genes putatively involved in PAH tolerance (MtSOD, MtPOX, MtAPX, MtGST, MtTFIIS, and MtTdp1α), (ii) activities of antioxidant (SOD, POX) and detoxification (GST) enzymes, and (iii) H2O2 and the heavy PAH, benzo[a]pyrene (B[a]P) accumulation. In the presence of B[a]P, whereas induction of the enzymatic activities was detected in R. irregularis and non-mycorrhizal roots as well as upregulation of the gene expressions in the non-mycorrhizal roots, downregulation of the gene expressions and decrease of enzyme activities were observed in mycorrhizal roots. Moreover, B[a]P increased H2O2 production in non-mycorrhizal roots and in R. irregularis but not in mycorrhizal roots. In addition, a lower B[a]P bioaccumulation in mycorrhizal roots was measured in comparison with non-mycorrhizal roots. Being less affected by pollutant toxicity, mycorrhizal roots did not activate any defense mechanism either at the gene expression regulation level or at the enzymatic level.

Defence mechanisms associated with mycorrhiza-induced resistance in wheat against powdery mildew.

To develop a more sustainable agriculture using alternative control strategies, mechanisms involved in the biocontrol ability of the arbuscular mycorrhizal fungus Funneliformis mosseae to protect wheat against the foliar biotrophic pathogen Blumeria graminis f. sp. tritici were investigated under controlled conditions. B. graminis infection on wheat leaves was reduced by 78% in mycorrhizal plants compared with non-mycorrhizal ones (control). Wheat roots inoculated with F. mosseae revealed a systemic resistance in leaves to B. graminis, after a 6-week co-culture. Accordingly, this resistance was associated with a significant reduction of B. graminis haustorium formation in epidermal leaf cells of mycorrhizal wheat and an accumulation of phenolic compounds and H2O2 at B. graminis penetration sites. Moreover, gene expression analysis demonstrated upregulation of genes encoding for several defence markers, such as peroxidase, phenylalanine ammonia lyase, chitinase 1 and nonexpressor of pathogenesis-related proteins 1 in mycorrhizal wheat only in the absence of the pathogen. This study showed that protection of wheat obtained against B. graminis in response to mycorrhizal inoculation by F. mosseae could be interpreted as a mycorrhiza-induced resistance (MIR). Our findings also suggest that MIR-associated mechanisms impaired the B. graminis development process and corresponded to a systemic elicitation of plant defences rather than a primed state in wheat leaves.

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.

Polyaromatic hydrocarbons impair phosphorus transport by the arbuscular mycorrhizal fungus Rhizophagus irregularis.

Phosphate uptake by plant roots is mainly mediated by arbuscular mycorrhizal fungi (AMF). However, the impact on phosphorus (P) transport of polycyclic aromatic hydrocarbons (PAH), persistent organic pollutants widely found in altered soils, is not known up today. Here, we monitored the Rhizophagus irregularis fungal growth and the fungal P transport ability from the extraradical mycelium to the host transformed chicory roots in the presence of anthracene and benzo[a]pyrene (B[a]P) and the combination of both PAH, under in vitro conditions. Firstly, our findings showed that PAH have detrimental effect on the fungal growth. The combination of both PAH was more toxic than each of the PAH individually due to synergistic effects. Secondly, PAH affected the P transport by the fungus from the medium to the roots. This was evidenced by either the decrease in (33)P quantity transported in the roots as well as the decrease in acid phosphatase activity in the mycorrhizal roots. Moreover, the fungal alkaline phosphatase activities remained constant in the extraradical mycelium as well as in the roots in the absence and in the presence of PAH. The GintPT and GiALP (encoding a P transporter and an alkaline phosphatase respectively) gene expressions were also found to be similar in the extraradical mycelium treated with PAH or not (control). These findings suggested that the P uptake by R. irregularis was not affected by PAH but probably the transport from the extraradical mycelium to the intraradical mycelium.

Role of arbuscular mycorrhizal symbiosis in root mineral uptake under CaCO3 stress.

This study investigated the effects of increasing CaCO(3) concentrations (0, 5, 10, 20 mM) on arbuscular mycorrhizal (AM) symbiosis establishment as well as on chicory root growth and mineral nutrient uptake in a monoxenic system. Although CaCO(3) treatments significantly decreased root growth and altered the symbiosis-related development steps of the AM fungus Rhizophagus irregularis (germination, germination hypha elongation, root colonization rate, extraradical hyphal development, sporulation), the fungus was able to completely fulfill its life cycle. Even when root growth decreased more drastically in mycorrhizal roots than in non-mycorrhizal ones in the presence of high CaCO(3) levels, the AM symbiosis was found to be beneficial for root mineral uptake. Significant increases in P, N, Fe, Zn and Cu concentrations were recorded in the mycorrhizal roots. Whereas acid and alkaline phosphatase enzymatic activities remained constant in mycorrhizal roots, they were affected in non-mycorrhizal roots grown in the presence of CaCO(3) when compared with the control.

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.

Genetic diversity and population structure in French populations of Mycosphaerella graminicola.

Mycosphaerella graminicola populations were examined in France with microsatellite markers and PCR-SSCP analysis of partial actin and ß-tubulin encoding sequences. A total of 363 isolates was sampled in 2005 from 17 provinces, and genotypes from corresponding strains were characterized. Unique haplotypes comprised 84% of the population, and gene diversity was high nationwide (0.70) and locally. A moderate genetic differentiation (G(ST) = 0.18) was found and indicated that in France the M. graminicola population was more structured than in other previously studied European countries. Bayesian structure analysis identified three genetic clusters distributed among the 17 provinces. Our results highlighted the potential for the adaptation of the fungus to local conditions, leading to genetic clusters among the French population of the fungus as well as genotype flow between regional clusters.

Mating type idiomorphs from a French population of the wheat pathogen Mycosphaerella graminicola: widespread equal distribution and low but distinct levels of molecular polymorphism.

Septoria tritici blotch caused by the heterothallic ascomycete Mycosphaerella graminicola is currently the most frequent and the most economically damaging disease on wheat worldwide. Five hundred and ten strains of this fungus were sampled from 16 geographical locations representing the major wheat producing areas in France. Multiplex PCR amplification, PCR-RFLP-SSCP screening and sequencing of parts of mating type encoding sequences were performed in order to assess the distribution and molecular polymorphism of the mating type idiomorphs. The two idiomorphs were scored at similar frequencies within all sampled locations. Both mating types were also identified at the leaf spatial scale, on 42% of leaves from which two or three strains were isolated. No correlation was found between distribution of mating types and either host cultivars from which the sampling was carried out or in vitro colony phenotypes observed during the culture of strains on potato dextrose agar (PDA) medium. PCR-RFLP-SSCP assay highlighted only one MAT1-1 strain exhibiting a profile distinct from all other MAT1-1 strains, whereas ten MAT1-2 strains (among which two and four with same profiles, respectively) showed profiles differing from the other MAT1-2 strains. Sequencing revealed that all polymorphisms corresponded to single nucleotide variations and all strains displaying the same single strand conformation polymorphism (SSCP) profiles showed identical nucleotide sequences, thereby confirming the high sensitivity of SSCP. Only two out of the disclosed nucleotide variations were nonsynonymous. This study strongly suggests a large potential for sexual reproduction in the French population of M. graminicola and reports a high conservation of mating type sequences in the fungus at both nucleotide and population levels, with a great difference in molecular variability between the two idiomorphs.