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International audience; Ulvan is a water-soluble sulfated heteropolysaccharide extracted from the cell walls of the green seaweeds Ulva spp. This polysaccharide is known to induce resistance and protect plants against a broad range of plant pathogenic fungi, such as Blumeria graminis f. sp. tritici (Bgt), the causal agent of powdery mildew in wheat. We aimed to study the defense mechanisms induced by ulvan against Bgt in susceptible wheat by investigating the defense-related gene expression, enzymes activity, accumulation of phenolic compounds and hydrogen peroxide (H2O2), as well as the development of Bgt infection structures in vitro and in planta. Symptoms were reduced by 42% in ulvan-treated plants. In vitro, ulvan did not inhibit conidial germination of Bgt but in planta, increased the appressorial germ tubes without haustorium. Ulvan increased the presence of fluorescent papillae and accumulation of H2O2 at the penetration sites of Bgt, as well as the content of phenolic compounds. POX, PAL and LOX activities were stimulated in ulvan-treated plants during the first 48 h after inoculation. However, few of defense-related genes studied were differentially expressed in infected plants after ulvan treatment. By contrast, in non-infected conditions, ulvan up-regulated the expression of genes involved in phenylpropanoid metabolism, i.e. PAL, CHS, COMT, ANS and FLS, genes encoding pathogenesis-related proteins, i.e. PR1, PR9, PR15, and LOX during the first 96 h after treatment. This study provides new insights about the multiple ulvan effects on wheat defense responses, and especially the elicitation of the phenylpropanoid pathway leading to phenolic compounds accumulation, which could be involved in cell wall reinforcement.

International audience; The present study aimed to evaluate the potential of the laminarin-based formulation Vacciplant to protect and induce resistance in wheat against Zymoseptoria tritici, a major pathogen on this crop. Under greenhouse conditions, a single foliar spraying of the product 2 days before inoculation with Z. tritici reduced disease severity and pycnidium density by 42 and 45%, respectively. Vacciplant exhibited a direct antifungal activity on Z. tritici conidial germination both in vitro and in planta. Moreover, it reduced in planta substomatal colonization as well as pycnidium formation on treated leaves. Molecular investigations revealed that Vacciplant elicits but did not prime the expression of several wheat genes related to defense pathways, including phenylpropanoids (phenylalanine ammonia-lyase and chalcone synthase), octadecanoids (lipoxygenase and allene oxide synthase), and pathogenesis‐related proteins (β‐1,3‐endoglucanase and chitinase). By contrast, it did not modulate the expression of oxalate oxidase gene involved in the reactive oxygen species metabolism. Ultrahigh-performance liquid chromatography-mass spectrometry analysis indicated limited changes in leaf metabolome after product application in both noninoculated and inoculated conditions, suggesting a low metabolic cost associated with induction of plant resistance. This study provides evidence that the laminarin-based formulation confers protection to wheat against Z. tritici through direct antifungal activity and elicitation of plant defense-associated genes.

International audience; Blumeria graminis f.sp. tritici (Bgt) is an obligate biotrophic fungal pathogen responsible for powdery mildew in bread wheat (Triticum aestivum L.). Upon Bgt infection, the wheat plant activates basal defense mechanisms namely PAMP-triggered immunity (PTI) in the leaves during the first few days. Understanding this early stage of quantitative resistance is crucial for developing new breeding tools and evaluating plant resistance inducers for sustainable agricultural practices. In this sense, we used a combination of transcriptomic and metabolomic approaches to analyze the early steps of the interaction between Bgt and the moderately susceptible wheat cultivar Pakito. Bgt infection resulted in an increasing expression of genes encoding pathogenesis-related proteins (PR-proteins, PR1, PR4, PR5 and PR8), known to target the pathogen, during the first 48 hours post-inoculation. Moreover, RT-qPCR and metabolomic analyses pointed out the importance of the phenylpropanoid pathway in quantitative resistance against Bgt. Among metabolites linked to this pathway, hydroxycinnamic acid amides containing agmatine and putrescine as amine component accumulated from the second to the fourth day after inoculation. This suggests their involvement in quantitative resistance via cross-linking processes in cell wall for reinforcement, what is supported by the up-regulation of PAL (phenylalanine ammonia-lyase), PR15 (encoding an oxalate oxidase) and POX (peroxidase) after inoculation. Finally, pipecolic acid, which is considered as a signal involved in systemic acquired resistance (SAR), accumulated after inoculation. These new insights lead to a better understanding of basal defense in wheat leaves after Bgt infection.

Plant resistance inducers are among the most promising alternatives to develop sustainable crop protection. Here, we examined the ability of saccharin, a metabolite derived from probenazole, to protect wheat against Zymoseptoria tritici, the most frequently occurring and damaging foliar pathogen on this crop. The experiments were performed in the greenhouse by treating seedlings of the wheat cultivar ‘Alixan’ with 15 mM of saccharin 2 days before challenge inoculation with the Z. tritici pathogenic strain T02596. Foliar application of saccharin resulted in 77% lower disease severity than in nontreated control plants. In vitro and in planta assays showed that saccharin did not exhibit any direct antifungal effect on spore germination or hyphal growth. Molecular investigations from 2 to 7 days posttreatment (dpt) revealed that saccharin treatment upregulates the expression of genes encoding for lipoxygenase (LOX) at all sampled time points and pathogenesis-related protein 1 (PR1) at 7 dpt, in both noninfectious and infectious contexts, as well as peroxidase (POX2) in noninfectious conditions. However, saccharin did not induce significant change in the expression of PAL gene encoding for phenylalanine ammonia-lyase. Our findings report for the first time the potential of saccharin to confer protection in wheat against Z. tritici through an elicitation and priming of LOX and PR gene-related defense pathways. Additional investigations would provide a better deciphering of defense mechanisms activated by this molecule in wheat against Z. tritici.

The hemibiotrophic fungus Zymoseptoria tritici, responsible for Septoria tritici blotch, is currently the most devastating foliar disease on wheat crops worldwide. Here, we explored, for the first time, the ability of rhamnolipids (RLs) to control this pathogen, using a total of 19 RLs, including a natural RL mixture produced by Pseudomonas aeruginosa and 18 bioinspired RLs synthesized using green chemistry, as well as two related compounds (lauric acid and dodecanol). These compounds were assessed for in vitro antifungal effect, in planta defence elicitation (peroxidase and catalase enzyme activities), and protection efficacy on the wheat-Z. tritici pathosystem. Interestingly, a structure-activity relationship analysis revealed that synthetic RLs with a 12 carbon fatty acid tail were the most effective for all examined biological activities. This highlights the importance of the C12 chain in the bioactivity of RLs, likely by acting on the plasma membranes of both wheat and Z. tritici cells. The efficacy of the most active compound Rh-Est-C12 was 20-fold lower in planta than in vitro, an optimization of the formulation is thus required to increase its effectiveness. No Z. tritici strain-dependent activity was scored for Rh-Est-C12 that exhibited similar antifungal activity levels towards strains differing in their resistance patterns to demethylation inhibitor fungicides, including multi-drug resistance strains. This study reports new insights into the use of bio-inspired RLs to control Z. tritici.

This study aimed to examine the ability of ulvan, a water-soluble polysaccharide from the green seaweed Ulva fasciata, to provide protection and induce resistance in wheat against the hemibiotrophic fungus Zymoseptoria tritici. Matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) analysis indicated that ulvan is mainly composed of unsaturated monosaccharides (rhamnose, rhamnose-3-sulfate, and xylose) and numerous uronic acid residues. In the greenhouse, foliar application of ulvan at 10 mg.ml–1 2 days before fungal inoculation reduced disease severity and pycnidium density by 45 and 50%, respectively. Ulvan did not exhibit any direct antifungal activity toward Z. tritici, neither in vitro nor in planta. However, ulvan treatment significantly reduced substomatal colonization and pycnidium formation within the mesophyll of treated leaves. Molecular assays revealed that ulvan spraying elicits, but does not prime, the expression of genes involved in several wheat defense pathways, including pathogenesis-related proteins (β-1,3-endoglucanase and chitinase), reactive oxygen species metabolism (oxalate oxidase), and the octadecanoid pathway (lipoxygenase and allene oxide synthase), while no upregulation was recorded for gene markers of the phenylpropanoid pathway (phenylalanine ammonia-lyase and chalcone synthase). Interestingly, the quantification of 83 metabolites from major chemical families using ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) in both non-infectious and infectious conditions showed no substantial changes in wheat metabolome upon ulvan treatment, suggesting a low metabolic cost associated with ulvan-induced resistance. Our findings provide evidence that ulvan confers protection and triggers defense mechanisms in wheat against Z. tritici without major modification of the plant physiology.

Keeping up agricultural production, while reducing chemical inputs, under biotic and abiotic stresses, falls within a considerable challenge. Among the solutions, developing more sustainable strategies to protect crops by using biocontrol agents is a promising alternative. Several beneficial microorganisms can limit pathogen development by direct (antibiosis, competition) and/or indirect effects (plant growth promotion and resistance induction) thanks to nutritional and hormonal balance regulations. A great deal of research articles have already reported the beneficial roles of rhizobacteria or arbuscular mycorrhizal fungi (AMF) on reducing the development of fungal pathogens when applied alone, or on plant nutrition when co-inoculated; while fewer of them reviewed the co-inoculation as a mean to protect against fungal pathogens. Hence, this review aims to present recent developments on the effectiveness of the co-inoculation with beneficial bacteria and AMF for fungal disease management. Firstly, main mechanisms beneath the beneficial effects of bacterial or AMF inoculation alone on plants are recalled. Secondly, results relating their combined inoculation on disease severity are presented and sorted according to their outcomes (synergistic, neutral, or antagonistic interactions), along with the possible underlying mechanisms. Finally, we gathered the main methodologies employed in tripartite interaction experiment and brought to light future challenges for practical use.

International audience; Alternative control strategies are increasingly encouraged to develop sustainable crop protection. In this aim, we assessed the ability of Dalgin Active ® , an Ascophyllum nodosum extract-based product, to induce resistance in bread wheat and durum wheat against Zymoseptoria tritici, a major fungal pathogen on these crops. Foliar application of the product provided a strong and significant reduction of disease intensity on both wheat species without any direct effect against the pathogen. Infection process monitoring revealed that Dalgin Active ® did not prevent fungal epiphytic growth and leaf colonization, but its application results in an inhibition of sporulation as well as fungal cell wall-degrading enzyme and protease activities. During the early stages of infection, Dalgin Active ® activated several plant defense markers on both wheat species, including PR protein, antioxidant metabolism, phenylpropanoid, and octadecanoid-based pathways. Although few differences were recorded, the induced defense reaction patterns were overall similar in both wheat species, suggesting that Dalgin Active ® could be used to biocontrol Z. tritici on both crops.

Plant resistance inducers are among the most promising alternatives to develop sustainable crop protection. Here, we examined the ability of saccharin, a metabolite derived from probenazole, to protect wheat against

To promote sustainable agriculture and healthy food, research that contributes towards a new generation of eco-friendly phytosanitary compounds is increasingly encouraged. The plant hormone salicylic acid (SA) is known for its ability to induce resistance in plants against a wide range of pathogens, whereas pyroglutamic acid (PGA), a constrained analogue of γ-aminobutyric acid, has never been studied in the context of plant protection.; Results: The present study investigated for the first time the protection efficacy of SA and PGA and five new conjugated derivatives against Zymoseptoria tritici, the main pathogen in wheat crops. SA and four derivatives showed significant disease severity reductions in planta (up to 49%). In vitro assays revealed that some molecules, including SA, displayed a small direct antifungal activity, whereas others, such as PGA, showed no effect. This finding suggests that, especially for molecules without any direct activity, the mode of action relies mainly on the induction of plant resistance.; Conclusion: Further investigations are needed to identify the defence pathways involved in plant resistance mechanisms elicited or primed by the molecules. The manufacture of these products was easily achieved on a scale of tens of grams of raw materials, and is easily scalable. The synthetic pathway is simple, short and inexpensive. For all of these reasons, the production of the target molecules is attractive for producers, whereas the prospect of a generation of non-polluting compounds with lasting efficiency against Z. tritici in wheat comes at a key moment for the sustainability of agriculture. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

International audience; 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 (gamma 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.

Bread wheat (BW) and durum wheat (DW) are both strongly affected by Septoria tritici blotch caused by the hemibiotrophic fungus Zymoseptoria tritici. However, only the BW-Z. tritici pathosystem has been well studied so far. Here, we compared compatible interactions between Z. tritici and both BW and DW species at the cytological, biochemical and molecular levels. Fungal infection process investigations showed close spore germination and leaf penetration features in both interactions, although differences in the patterns of these events were observed. During the necrotrophic phase, disease severity and sporulation levels were associated in both interactions with increases of the two cell-wall degrading enzyme activities endo-β-1,4-xylanase and endo-β-1,3-glucanase as well as protease. An analysis of plant defense responses during the first five days post inoculation revealed inductions of GLUC, Chi4, POX and PAL and a repression of LOX gene expressions in both wheat species, although differences in kinetics and levels of induction or repression were observed. In addition, peroxidase, catalase, glucanase, phenylalanine ammonia-lyase and lipoxygenase activities were induced in both wheat species, while only weak accumulations of hydrogen peroxide and polyphenols were detected at the fungal penetration sites. Our study revealed overall a similarity in Z. tritici infection process and triggered wheat defense pathways on both pathosystems.

Trunk diseases have become among the most important grapevine diseases worldwide. They are caused by fungal pathogens that attack the permanent woody structure of the vines and cause various symptoms in woody and annual organs. This study examined modifications of plant responses in green stem, cordon, and trunk of grapevines expressing Esca proper (E) or apoplexy (A) event, which are the most frequent grapevine trunk disease symptoms observed in Europe. Transcript expression of a set of plant defense- and stress-related genes was monitored by quantitative reverse-transcription polymerase chain reaction while plant phytoalexins and fungal metabolites were quantified by high-performance liquid chromatography-mass spectrometry in order to characterize the interaction between the grapevine and trunk disease agents. Expression of genes encoding enzymes of the phenylpropanoid pathway and trans-resveratrol content were altered in the three organs of diseased plants, especially in the young tissues of A plants. Pathogenesis-related proteins and the antioxidant system were severely modulated in A plants, which indicates a drastic stress effect. In the meantime, fungal polyketides 6-MSA, (R)-mellein, and (3R,4R)-4-hydroxymellein, were accumulated in A plants, which suggests their potential effect on plant metabolism during the appearance of foliar symptoms.

International audience; A potential alternative strategy to chemical control of plant diseases could be the stimulation of plant defense by arbuscular mycorrhizal fungi (AMF). In the present study, the influence of three parameters (phosphorus supply, mycorrhizal inoculation, and wheat cultivar) on AMF protective efficiency against Blumeria graminis f. sp. tritici, responsible for powdery mildew, was investigated under controlled conditions. A 5-fold reduction (P/5) in the level of phosphorus supply commonly recommended for wheat in France improved Funneliformis mosseae colonization and promoted protection against B. graminis f. sp. tritici in a more susceptible wheat cultivar. However, a further decrease in P affected plant growth, even under mycorrhizal conditions. Two commercially available AMF inocula (F. mosseae, Solrize®) and one laboratory inoculum (Rhizophagus irregularis) were tested for mycorrhizal development and protection against B. graminis f. sp. tritici of two moderately susceptible and resistant wheat cultivars at P/5. Mycorrhizal levels were the highest with F. mosseae (38 %), followed by R. irregularis (19 %) and Solrize® (SZE, 8 %). On the other hand, the highest protection level against B. graminis f. sp. tritici was obtained with F. mosseae (74 %), followed by SZE (58 %) and R. irregularis (34 %), suggesting that inoculum type rather than mycorrhizal levels determines the protection level of wheat against B. graminis f. sp. tritici. The mycorrhizal protective effect was associated with a reduction in the number of conidia with haustorium and with an accumulation of polyphenolic compounds at B. graminis f. sp. tritici infection sites. Both the moderately susceptible and the most resistant wheat cultivar were protected against B. graminis f. sp. tritici infection by F. mosseae inoculation at P/5, although the underlying mechanisms appear rather different between the two cultivars. This study emphasizes the importance of taking into account the considered parameters when considering the use of AMF as biocontrol agents.

The rhizospheric Bacillus subtilis PTA-271 (271) and endophytic Pseudomonas fluorescens PTA-CT2 (CT2) and Pantoea agglomerans PTA-AF2 (AF2) bacteria are able to induce systemic resistance (ISR) in grapevine against B. cinerea, but ISR markers and their costs remained unknown in vineyards. In this study, we investigated the relationship between the effectiveness of single and binary combinations of selected bacteria to induce ISR and their ability to trigger phytoalexin accumulation, as a potential marker for disease resistance, in leaves and berries, as well as their impact on grape yield in vineyards. Grapevine plants were treated during 2006 in two vineyards by drenching soil with single or binary mixtures of bacteria. Induced resistance against B. cinerea was evaluated and stilbenic phytoalexins were analyzed by HPLC in both leaves and berries. Grape yield was also assessed as number and weight of clusters at ripening. Both single and mixtures of bacteria were effective in reducing gray mold severity in the leaves and berries in vineyards. Disease control was accompanied by a significant accumulation of stilbenic phytoalexins, trans-resveratrol and e-viniferin, in both leaves and berries in the bacterized plants. δ-Viniferin also accumulated, but only in berries of the treated plants. Reduction of disease symptoms and accumulation of resveratrol and viniferins were higher in the plants treated with single CT2 compared to AF2 and 271. Treatment of grapevine plants with binary mixtures of these isolates resulted in a significant performance of CT2+AF2 in leaves and CT2+271 in berries. On the other hand, bacterial treatments did not show any negative effect on grape yield. These results revealed the efficacy of CT2 alone or in combination with AF2 or with 271 in triggering grapevine resistance against B. cinerea and enhancing systemic accumulation of resveratrol and viniferins, without compromising grape yield.

International audience; Little is known about the influence of arbuscular mycorrhizal fungi (AMF) inoculum sources on phytoremediation efficiency. Therefore, the aim of this study was to compare the effects of two mycorrhizal inocula (indigenous and commercial inocula) in association with alfalfa and tall fescue on the plant growth, the bacterial, fungal, and archaeal communities, and on the removal of dioxin/furan (PCDD/F) from a historically polluted soil after 24 weeks of culture in microcosms. Our results showed that both mycorrhizal indigenous and commercial inocula were able to colonize plant roots, and the growth response depends on the AMF inoculum. Nevertheless, the improvement of root dry weight in inoculated alfalfa with indigenous inoculum and in inoculated tall fescue with commercial inoculum was clearly correlated with the highest mycorrhizal colonization of the roots in both plant species. The highest shoot dry weight was obtained in inoculated alfalfa and tall fescue with the commercial inoculum. AMF inoculation differently affected the number of bacterial and archaeal OTUs and bacterial diversity, with elevated bacterial and archaeal OTUs and bacterial diversity observed with indigenous inoculum. Mycorrhizal inoculation increases the abundance of bacterial OTUs (in particular with indigenous inoculum) and microbial richness but it does not improve PCDD/F dissipation. Vegetation had no effect on the abundance of microbial OTUs nor on richness but stimulated specific communities (Planctomycetia and Gammaproteobacteria) likely to be involved in the dissipation of PCDD/F. The reduction of toxic equivalency PCDD/F concentration also could be explained by the stimulation of soil microbial activities estimated with dehydrogenase and fluorescein diacetate hydrolase.

International audience; 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.

To promote sustainable agriculture and healthy food, research that contributes towards a new generation of eco-friendly phytosanitary compounds is increasingly encouraged. The plant hormone salicylic acid (SA) is known for its ability to induce resistance in plants against a wide range of pathogens, whereas pyroglutamic acid (PGA), a constrained analogue of γ-aminobutyric acid, has never been studied in the context of plant protection.The present study investigated for the first time the protection efficacy of SA and PGA and five new conjugated derivatives against Zymoseptoria tritici, the main pathogen in wheat crops. SA and four derivatives showed significant disease severity reductions in planta (up to 49%). In vitro assays revealed that some molecules, including SA, displayed a small direct antifungal activity, whereas others, such as PGA, showed no effect. This finding suggests that, especially for molecules without any direct activity, the mode of action relies mainly on the induction of plant resistance.Further investigations are needed to identify the defence pathways involved in plant resistance mechanisms elicited or primed by the molecules. The manufacture of these products was easily achieved on a scale of tens of grams of raw materials, and is easily scalable. The synthetic pathway is simple, short and inexpensive. For all of these reasons, the production of the target molecules is attractive for producers, whereas the prospect of a generation of non-polluting compounds with lasting efficiency against Z. tritici in wheat comes at a key moment for the sustainability of agriculture. © 2018 Society of Chemical Industry.

International audience; Plant resistance inducers, also referred to as elicitors, are agents that confer improved protection to pathogen or pest attacks by inducing host defense mechanisms. Such products are effective against a wide range of crop enemies, including viruses, bacteria, fungi, oomycetes, nematodes, and herbivores. The mode of action of these products differs from that of traditional pesticides because they do not target directly the bio-aggressor through antifungal activity, but they inhibit its development indirectly via the elicitation of plant defense reactions. In the current context of sustainable agriculture and growing demand for healthy food, plant resistance inducers are considered as an eco-friendly and promising alternative to conventional pesticides, and their implementation in integrated pest management strategy is strongly encouraged. Plant resistance inducers can be of synthetic or natural origin. This chapter will focus on resistance inducers of natural origin including living microorganisms, plant extracts, microbial cell-wall extracts, microbial metabolites, minerals, and ions. An overview on the market and recent advances on the regulation of these products as well as future challenges to promote their development and wide use in disease management programs will be described.

International audience; We investigated here for the first time the infectious process and CWDE activities of Zymoseptoria tritici on durum wheat (DW), by using three Tunisian cultivars with different susceptibility levels to the pathogen: Karim (susceptible), Nasr (moderately resistant) and Salim (resistant). High DW resistance level was associated with decreased direct leaf penetration at early stage of infection and with reduced sporulation and production of CWDE (endo-b-1,4-xylanase and endo-b-1,3-glucanase) and protease activities during the necrotrophic phase. Furthermore, the expression pattern of six wheat defenserelated genes, including GLUC, Chi 4, POX, Msr and Bsi1, exhibited significant differences between resistant and susceptible DW cultivars. Altogether, these results provide new insight into the nonspecific resistance mechanisms in durum wheat against Z. tritici.

International audience; In the context of an increasing worldwide food requirement, the control of crop diseases is crucial to guarantee high and stable yield, as well as sanitary quality. An environmentally friendly contribution to this could be biocontrol using beneficial microorganisms, such as arbuscular mycorrhizal fungi (AMF). AMF establish symbiosis with their host plants, thus influencing their growth, but they also induce tolerance to environmental stresses. Among stresses that can be alleviated through AMF inoculation, plant attacks by aerial pathogens and pests have so far been underestimated. Therefore, we present here an overview of studies focusing on AMF-mediated bioprotection against aerial pathogens and pests. Obtained protection is mainly due to changes in host nutrition and induction of defense following the establishment of arbuscular mycorrhizal symbiosis. This protection can vary greatly depending on different factors such as host genotype, AMF species involved, pest and pathogen lifestyles, interactions between AMF and other microorganisms, or even crop management practices. Finally, some future challenges for the use of AMF in biocontrol are discussed.

IPM SPE UB; International audience; Esca disease is one of the major grapevine trunk diseases in Europe and the etiology is complex, since several inhabiting fungi are identified to be associated with this disease. Among the foliar symptom expressions, the apoplectic form may be distinguished and characterized by sudden dieback of shoots, leaf drop, and shriveling of grape clusters in a few days that can ultimately induce the plant death. To further understand this drastic event, we conducted transcriptomic and metabolomic analyses to characterize responses of leaves during the period preceding symptom appearance (20 and 7 days before foliar symptom expression) and at the day of apoplexy ex- pression. Transcriptomic and metabolomic analyses provide signatures for the apoplectic leaves and most changes concerning the metabolism of carbohydrates, amino acids, and phenyl- propanoids. In deciphering glutathione-S-transferase (GST), its preferential location in phloem, correlated with the upregulation of GST genes and a decrease of the glutathione level, offers further support to the putative role of glutathione during apo- plexy expression.

Botryosphaeria dieback is a fungal grapevine trunk disease that currently represents a threat for viticulture worldwide because of the important economical losses due to reduced yield of affected plants and their premature death. Neofusicoccum parvum and Diplodia seriata are among the causal agents. Vine green stems were artificially infected with N. parvum or D. seriata at the onset of three different phenological stages (G stage (separated clusters), flowering and veraison). Highest mean lesion lengths were recorded at flowering. Major proteome changes associated to artificial infections during the three different phenological stages were also reported using two dimensional gel electrophoresis (2D)-based analysis. Twenty (G stage), 15 (flowering) and 13 (veraison) differentially expressed protein spots were subjected to nanoLC-MS/MS and a total of 247, 54 and 25 proteins were respectively identified. At flowering, a weaker response to the infection was likely activated as compared to the other stages, and some defense-related proteins were even down regulated (e.g., superoxide dismutase, major latex-like protein, and pathogenesis related protein 10). Globally, the flowering period seemed to represent the period of highest sensitivity of grapevine to Botryosphaeria dieback agent infection, possibly being related to the high metabolic activity in the inflorescences.

International audience; 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.

Botryosphaeria dieback is a grapevine trunk disease with a worldwide distribution associated with Diplodia seriata and Neofusicoccum parvum, among several other Botryosphaeriaceae species. The aforementioned xylem-inhabiting fungi cause wood lesions and leaf and berry symptoms, and eventually lead to the death of the plant. The aim of this work was to develop a simple model system to reproduce the foliar symptoms caused by D. seriata and N. parvum to better characterize fungal pathogenicity and determine the mechanisms involved in symptom development. Green stems of grafted ‘Aragonez’ grapevine cuttings were inoculated with three isolates of N. parvum and two isolates of D. seriata with different degrees of virulence and the experiment was repeated four times from 2011 to 2014. Three months after inoculation, the lesions associated with N. parvum were larger than those associated with D. seriata. Similarly, 8 months after inoculation, the percentage of plants showing foliar symptoms was greater in the N. parvum treatments than in the D. seriata treatments. During the emergence of foliar symptoms, plant stress-related responses were modulated in green stems and leaves, especially a downregulation of superoxide dismutase (SOD) and fasciclin-like arabinogalactan protein (fascAGP) and an upregulation of stilbene synthase (STS) genes with an accumulation of phenolics. In conclusion, the simple model system developed allowed a good characterization of isolate pathogenicity and correlation with foliar symptoms of Botryosphaeria dieback, namely spots on leaf margin and blade.

This review presents an overview of eutypa dieback, esca and botryosphaeria dieback, the predominant grapevine trunk diseases worldwide. It covers their symptomatologies in the trunk, leaves and berries; the characteristics of the different fungal species associated with them; and host‐pathogen interactions. Here, the host‐pathogen relationship is defined at the cytological, physiological and molecular levels. Currently available experimental tools for studying these diseases, both in vitro and in the field, are discussed. Finally, a progress report on their control, which, since the ban of sodium arsenite, comprises chemical, biological and⁄or sanitation methods, is presented.

Previous experiments showed that seven bacteria identified as Acinetobacter lwoffii (PTA-113 and PTA-152), Bacillus subtilis (PTA-271), Pantoea agglomerans (PTA-AF1 and PTA-AF2) and Pseudomonas fluorescens (PTA-268 and PTA-CT2) induced systemic resistance in grapevine against Botrytis cinerea. Based on these findings, we investigated biocontrol capacity of different mixtures under vineyard conditions over three consecutive years. Treatments with bacterial mixtures were shown to induce systemic resistance against B. cinerea on year 2. Efficacy and duration of such a disease control seemed to be reinforced on year three without renewal of bacterial treatments. Accordingly, the effectiveness of induced resistance varied with mixture type of bacteria and was accompanied by a stimulation of chitinase and β-1,3 glucanase activities in both leaves and berries. Interestingly, treatments with mixtures containing both selected strains of P. agglomerans (PTA-AF1 + PTA-AF2) appeared as the most effective in triggering systemically the plant defense reactions and reducing the symptoms of grey mould disease.

In this study, the biocontrol ability of seven grapevine-associated bacteria, previously reported as efficient against Botrytis cinerea under in vitro conditions, was evaluated in two vineyard orchards with the susceptible cv. Chardonnay during four consecutive years (2002–2005). It was shown that the severity of disease on grapevine leaves and berries was reduced to different levels, depending on the bacterial strain and inoculation method. Drenching the plant soil with these bacteria revealed a systemic resistance to B. cinerea, even without renewal of treatment. Accordingly, this resistance was associated with a stimulation of some plant defense responses such as chitinase and β-1,3-glucanase activities in both leaves and berries. In leaves, chitinase activity increased before veraison (end-July) while β-1,3-glucanase reached its maximum activity at ripening (September). Reverse patterns were observed in berries, with β-1,3-glucanase peaking at full veraison (end-August) and chitinase at a later development stage. Highest activities were observed with Acinetobacter lwoffii PTA-113 and Pseudomonas fluorescens PTA-CT2 in leaves, and with A. lwoffii PTA-113 and Pantoea agglomerans PTA-AF1 in berries. These results have demonstrated an induced protection of grapevine against B. cinerea by selected bacteria under field conditions, and suggest that induced resistance could be related to a stimulation of plant defense reactions in a successive manner.

Liquid chromatography-diode array screening of the organic extract of the cultures of 13 isolates of the fungus Neofusicoccum parvum, the main causal agent of botryosphaeria dieback of grapevine, showed similar metabolites. One strain was selected for further chemical studies and led to the isolation and characterisation of 13 metabolites. Structures were elucidated through spectroscopic analyses, including one- and two-dimensional NMR and mass spectrometry, and through comparison to literature data. The isolated compounds belong to four different chemical families: five metabolites, namely, ( )-terremutin (1), (+)-terremutin hydrate (2), (+)-epi-sphaeropsidone (3) ( )-4-chloro-terremutin hydrate (4) and(+)-4- hydroxysuccinate-terremutin hydrate (5), belong to the family of dihydrotoluquinones; two metabolites, namely, (6S,7R) asperlin (6) and (6R,7S)-dia-asperlin (7), belong to the family of epoxylactones; four metabolites, namely, (R)-( )-mellein (8), (3R,4R)-4-hydroxymellein (9), (3R,4S)-4-hydroxymellein (10) (R)( )-3-hydroxymellein (11), belong to the family of dihydroisocoumarins; and two of the metabolites, namely, 6-methyl-salicylic acid (12) and 2-hydroxypropyl salicylic acid (13), belong to the family of hydroxybenzoic acids. We determined the phytotoxic activity of the isolated metabolites through a leaf disc assay and the expression of defence-related genes in Vitis vinifera cells cv. Chardonnay cultured with ( )-terremutin (1), the most abundant metabolite. Finally, analysis of the brown stripes of grapevine wood from plants showing botryosphaeria dieback symptoms revealed the presence of two of the isolated phytotoxins info:eu-repo/semantics/publishedVersion

Sphingolipids are emerging as second messengers in programmed cell death and plant defense mechanisms. However, their role in plant defense is far from being understood, especially against necrotrophic pathogens. Sphingolipidomics and plant defense responses during pathogenic infection were evaluated in the mutant of long-chain base phosphate (LCB-P) lyase, encoded by the dihydrosphingosine-1-phosphate lyase1 (AtDPL1) gene and regulating long-chain base/LCB-P homeostasis. Atdpl1 mutants exhibit tolerance to the necrotrophic fungus Botrytis cinerea but susceptibility to the hemibiotrophic bacterium Pseudomonas syringae pv tomato (Pst). Here, a direct comparison of sphingolipid profiles in Arabidopsis (Arabidopsis thaliana) during infection with pathogens differing in lifestyles is described. In contrast to long-chain bases (dihydrosphingosine [d18:0] and 4,8-sphingadienine [d18:2]), hydroxyceramide and LCB-P (phytosphingosine-1-phosphate [t18:0-P] and 4-hydroxy-8-sphingenine-1-phosphate [t18:1-P]) levels are higher in Atdpl1-1 than in wild-type plants in response to B. cinerea. Following Pst infection, t18:0-P accumulates more strongly in Atdpl1-1 than in wild-type plants. Moreover, d18:0 and t18:0-P appear as key players in Pst- and B. cinerea-induced cell death and reactive oxygen species accumulation. Salicylic acid levels are similar in both types of plants, independent of the pathogen. In addition, salicylic acid-dependent gene expression is similar in both types of B. cinerea-infected plants but is repressed in Atdpl1-1 after treatment with Pst. Infection with both pathogens triggers higher jasmonic acid, jasmonoyl-isoleucine accumulation, and jasmonic acid-dependent gene expression in Atdpl1-1 mutants. Our results demonstrate that sphingolipids play an important role in plant defense, especially toward necrotrophic pathogens, and highlight a novel connection between the jasmonate signaling pathway, cell death, and sphingolipids.

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Among grapevine trunk diseases, esca proper and apoplexy commonly represent a threat for viticulture worldwide. To retrieve further information about the mechanisms activated in apoplectic and esca proper-affected plants, a two-dimensional gel electrophoresis (2-DE) based analysis was conducted on green stems from 26-year-old standing vines. Symptomatic and asymptomatic stems from both apoplectic (A) and esca proper-affected (E) plants compared to control (without visual symptom since 10 years) stems were studied. Thirty-three differentially expressed proteins were identified by nanoLC-MS/MS and included into three groups conceptually defined as proteins involved in (i) metabolism and energy, (ii) stress tolerance, and (iii) defense response. For nine of them, expression of the relative mRNA's was also monitored by qRT-PCR. Proteome variations were specifically related to apoplexy and esca proper but were more similar in asymptomatic stems than in the symptomatic ones. Remarkable quantitative differences were noted for several proteins in symptomatic stems according to the expressed form, A and E. Results further indicate that similar responses are likely activated in asymptomatic stems but a various quantitative expression is triggered upon onset of apoplexy or esca proper symptoms while both kind of plants are infected by the same pathogenic fungi.

Esca is a destructive disease in grapevines (Vitis vinifera L.) caused by at least three fungi and characterised by two different external symptoms, the apoplectic and leaf stripe form. This latter form can be discerned as soon as symptoms become visible, but the preceding discrete signs during incubation are poorly or not understood. To further understand the development of the leaf stripe form, the period preceding and following the appearance of symptoms was investigated by studying physiological and molecular markers associated with photosynthetic mechanisms and stress response. No perturbation of any targeted metabolism was observed in asymptomatic leaves of asymptomatic canes from vines showing the leaf stripe form of esca. Conversely, drastic alterations of photosynthesis functions were registered in presymptomatic leaves, as revealed by the decrease of gas exchange and chlorophyll fluorescence, and the repression of photosynthesis-related genes. These alterations were amplified during symptom development. Expression of defence-related genes was affected and detected early in presymptomatic leaves and amplified during symptom expression. Our results suggest that grapevines may react precociously by reducing photosynthesis and triggering defence mechanisms in response to the leaf stripe form of esca.

International audience; La septoriose, maladie causée par le champignon hémibiotrophe Zymoseptoria tritici, est l’une des maladies les plus dommageables pour la culture du blé. Dans un contexte de transition agroécologique, le développement des stratégies de lutte respectueuses de la santé humaine et de l’environnement est une priorité. La colonisation racinaire des plantes par les champignons mycorhiziens à arbuscules (CMA) est connue pour induire la MIR (Mycorrhiza Induced Resistance), correspondant à un état de résistance de la plante à divers bio-agresseurs. Nous avons ainsi testé l’efficacité du CMA Funneliformis mosseae à protéger le blé contre la septoriose dans le cadre du projet Bioscreen (Smartbiocontrol, Interreg V).Des plantules de blé tendre (cv. Alixan) sensibles à la septoriose ont été cultivées dans un substrat contenant le CMA F. mosseae. Après 6 semaines de culture, les parties aériennes des plantules ont été infectées avec une solution de spores de Z. tritici, puis les symptômes de la maladie ont été évalués 3 semaines après infection. Des feuilles et racines ont été prélevées pour une évaluation de l’expression génique. Des taux de protection de 78% et de 87% contre la septoriose ont été obtenus chez le blé mycorhizé. Cette protection au niveau foliaire a été corrélée à un effet éliciteur systémique induit par la mycorhization sur un ensemble de gènes de défense, également induits lors d’une attaque par Z. tritici, et codant des PR protéines (PR1, PR4 et PR5) ou des protéines impliquées contre le stress oxydatif (POX et WRKY53). Au niveau racinaire, des gènes marqueurs de la symbiose mycorhizienne impliqués dans les échanges de nutriments (AMT3.1, PT10, PT11, Sult, Kchan et ABC) étaient surexprimés chez le blé mycorhizé sans altération par la septoriose. Nos résultats montrent la mise en place d’une MIR chez le blé, renforçant ses défenses contre la septoriose.