Gaeumannomyces Graminis Var Research Articles

Multifunctional manganese-based carboxymethyl chitosan hydrogels for pH-triggered pesticide release and enhanced fungicidal activity

Carboxymethyl chitosan (CMCS) has emerged as a promising biopolymer carrier for controlled release formulations of pesticide. In this study, manganese-based carboxymethyl chitosan hydrogel was facilely prepared to encapsulate and release fungicide prothioconazole in a controllable manner. The loading content (LC) and encapsulation efficiency (EE) of prothioconazole were optimized by orthogonal test. When scaled up under the optimal condition, the corresponding LC and EE were 22.17 % and 68.38 %, respectively. The result showed that the pH-triggered release behavior of prothioconazole for the hydrogels was consistent with swelling behavior. The pesticide rapidly released in neutral and slightly alkaline solutions than in acidic conditions. Moreover, the prepared hydrogel showed enhanced fungicidal ability against wheat take-all pathogen (Gaeumannomyces graminis var. tritic) compared to that of prothioconazole technical material. This research seeks to provide a promising approach to develop metal and polysaccharide-based hydrogels to control the pesticide release and reduce pesticide use in sustainable agriculture application.

Isolation and yield optimization of lipopeptides from Bacillus subtilis Z-14 active against wheat take-all caused by Gaeumannomyces graminis var. tritici.

Wheat take-all, caused by the soil-borne fungus Gaeumannomyces graminis var. tritici, is one of the major constraints on wheat production worldwide. Bacillus subtilis Z-14 exerts significant biocontrol activity against wheat take-all, and lipopeptide antibiotics are the main antifungal substances. Herein, lipopeptide antibiotics C14-C15 iturin A, C14-C16 fengycin A, and C15-C17 fengycin B from B. subtilis Z-14 culture filtrates were separated and identified by high-performance liquid chromatography, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry, and mass spectrometry/mass spectrometry, respectively. The optimal medium components for Z-14 lipopeptide antibiotic production were 3.85g/L corn flour, 1.57g/L soybean meal, 0.03g/L FeSO4 ·7H2 O, 0.2g/L NaH2 PO4 ·2H2 O, and 0.4g/L Na2 HPO4 ·2H2 O. Quantification analysis by high-performance liquid chromatography showed that fengycins played a main role in antifungal activity against Gaeumannomyces graminis var. tritici. Quantitative reverse transcription polymerase chain reaction showed that lipopeptide synthesis genes fenD and ituC reached maximum expression levels after 48 h of fermentation. The strongest control of wheat take-all by Z-14 was achieved by adding 30mL of culture filtrate per 350g of soil in pot experiments, during which disease reduction reached 88.15%. This study provides theoretical support and a material basis for the prevention and treatment of wheat take-all disease.

Quantification of Pseudomonas protegens FD6 and Bacillus subtilis NCD-2 in soil and the wheat rhizosphere and suppression of root pathogenic Rhizoctonia solani AG-8

Bacillus subtilis strain NCD-2 and Pseudomonas protegens strain FD6 produce a range of antimicrobial metabolites known to be effective against soil-borne plant pathogens. Quantitative qPCR assays were designed from sequence characterized amplified fragment length polymorphisms (AFLP) diagnostic for each inoculant genotype. The qPCR minimum detection thresholds (MDT) for NCD-2 and FD6 ranged between 5.2 × 103 (log 3.71) and 1.5 × 106 (log 6.18) genome (cell) equivalents g−1 of soil or wheat root tissue and were independent of soil type. Wheat rhizosphere populations of strains FD6 and NCD-2 significantly declined during the first 14 days postinoculation and then remained stable for the next 14 days. Colonization of wheat roots, rhizosphere soil and bulk (non-rhizosphere) soil by FD6 was significantly greater than that of NCD-2. Strains FD6 and NCD-2 significantly decreased in vitro growth of wheat root pathogenic Rhizoctonia solani AG-8, Pythium irregulare and Gaeumannomyces graminis var. tritici, fungal antibiosis by FD6 greater than NCD-2. Wheat rhizosphere colonization by NCD-2 and FD6 significantly decreased abundance of R. solani AG-8 in the rhizosphere by 3- and 13-fold respectively, pathogen suppression increasing (21%–29%) foliar biomass of wheat seedlings to levels not significantly different to the non-diseased control. The qPCR assays developed in this study were used to monitor soil and rhizosphere populations of P. protegens FD6 and B. subtilis NCD-2 and their suppressive efficacies against R. solani AG-8 in wheat. These assays can be applied to monitor the dynamics of these inoculants in suppressing Rhizoctonia root rot and other crop diseases.

Influence of sulfur fertilization on infection of wheat Take-all disease caused by the fungus Gaeumannomyces graminis var. tritici

Influence of sulfur fertilization on infection of wheat Take-all disease caused by the fungus Gaeumannomyces graminis var. tritici

Antifungal activity of the lemongrass and clove oil encapsulated in mesoporous silica nanoparticles against wheat's take-all disease

Combined application of plant essential oils (EOs) with known antimicrobial effects and silica nanocapsules with high loading capacity and protection capability of the EOs make them proper candidates for creating environmentally friendly fungicides. In this study, EOs of the Lemongrass (LGO) and Clove (CO) were used against Gaeumannomyces graminis var. tritici (Ggt), a causal agent of take-all disease of wheat. To provide controlled delivery of the EOs, they were encapsulated into mesoporous silica nanoparticles (MSNPs) and then compared to the effects of pure EOs both in- vitro and in- vivo. MSNPs were synthesized via the sol-gel process. Various techniques such as Fourier transform infrared spectroscopy (FTIR), the Brunauer–Emmett–Teller (BET), thermogravimetric analysis (TGA), and UV–Vis spectroscopy were used to evaluate the successful loading of the EOs into the pore of MSNPs. The encapsulation efficiency (EE) was calculated as high as 84.24% for LGO and 80.69% for CO, while loading efficiency (LE) was determined 36% and 29% for LGO and CO, respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) displayed spherical shapes and porous structures with average diameters of 50–70 nm. Recognition of the main components of the EOs via gas chromatographic-mass spectrometry (GC–MS) before and after the EO loading, detected eugenol and citral as the most frequent compounds in LGO and CO, respectively. For antifungal test in- vitro, selected concentrations of the pure EOs, EOs loaded in MSNPs (EOs- MSNPs) and Mancozeb ® fungicide based on pre-tests were mixed using potato dextrose agar (PDA). The inhibition percentage (IP) of fungal growth at each concentration, as well as minimum inhibition concentration (MIC) and minimum fungicidal concentrations (MFC) were obtained. The results indicated that antifungal effects in the encapsulated form increased by up to three times. In- vivo, the sterile wheat seeds were treated with pure EOs, EOs-MSNPs, and mancozeb at MFC concentration. Also, in order to keep on the EOs-MSNPs around the seeds, sodium alginate was used. The consequences of in- vivo experiments indicated that rate of disease control in presence of EOs-MSNPs and mancozeb was the same (~70%) and higher than pure EOs (LGO: 57.44%, CO: 49%). Also, improving the growth parameters in wheat plant, the covering of the EOs-MSNPs in alginate, had better control (84%) than that of EOs-MSNPs alone. Further, the release kinetics studies showed a gradual release of LGO and CO from MSNPs for four weeks in water and for five weeks in the soil-plant system. To the best of our knowledge, this is the first report of the control effect of LGO, CO, and their nanocapsule in MSNPs against the take-all disease of wheat. These results showed that the EOs-MSNPs can be a safe product for the efficient control of take-all disease in wheat crop.

Wheat root transcriptional responses against Gaeumannomyces graminis var. tritici

Wheat root rot caused by Gaeumannomyces graminis var. tritici (Ggt) results in severe yield losses in wheat production worldwide. However, little is known about the molecular mechanism that regulates systemic symptom development in infected wheat. Fluorescent microscopy observation of the stained wheat roots infected by Ggt showed that lesions were visible when the fungus could be detected in the endodermis, pericycle and phloem at 5 days post inoculation (dpi), and rust symptoms were visible when there was extensive fungal colonization in the root cortex at 6 dpi. Transcriptome sequencing of Ggt-inoculated wheat roots and healthy control root samples was performed at 5 dpi to identify Ggt-induced gene expression changes in wheat roots at the time of lesion formation. A total of 3973 differentially expressed genes (DEGs) were identified, of which 1004 (25.27%) were up-regulated and 2969 (74.73%) were down-regulated in Ggt-inoculated wheat roots compared with those in control roots. GO annotation and KEGG pathway analysis of these DEGs revealed that many of them were associated with pathogen resistance, such as those involved in oxidation-reduction process, tryptophan biosynthesis process, and phenylpropanoid biosynthesis process. Analysis of DEGs revealed that 15 DEGs were involved in cellular regulation, 57 DEGs in signal transduction pathways, and 75 DEGs in cell wall reorganization, and 23 DEGs are pathogenesis-related proteins. Reverse transcription quantitative PCR (RT-qPCR) of 13 of those DEGs showed that these genes may play roles in wheat resistance against Ggt. Overall, this study represents the first transcriptional profiling of wheat roots in response to Ggt infection and further characterization of DEGs identified in this study may lead to better understanding of resistance against take-all in wheat.

Control of Take-all disease of wheat caused by Gaeumannomyces graminis var. tritici through micronutrients (Zn, Fe and Cu)

Take-all caused by the soil-born fungi Gaeumannomyces graminis (Sacc) Arx & Olivier is an important crown and root rot diseases of wheat throughout the world. Biological control and plant nutrition management are two effective methods for the control of this disease. In this study, effect of plant nutrition was investigated on improvement of wheat growth and suppression of the Take-all disease. For this purpose, the effect of iron sulfate, copper sulfate and zinc sulfate on Take-all disease of wheat was studies on the basis of infected roots and production of shoot dry weight. The obtained results revealed that the experimental soil was slightly alkaline. The soil contained adequate amounts of nitrogen and zinc, but was poor in availability of iron, cupper and mild deficit in phosphorus. In order to investigate the effect of the nutrients on growth and severity of the disease, four levels (0, 2.5, 5.0 and 10 mg/kg soil) of micronutrients were used. Iron had greatest effect on suppression of Take-all disease among applied micronutrients. The infected roots decreased from 89 to 53% and abolished symptoms on wheat crown up to nine times with the application if iron. In addition, iron increased the dry weight of aerial parts of wheat from 0.78 to 1.3 g per plant. The effects of zinc and copper fertilizers on disease suppression on wheat was less than that of iron with decreased infected roots to 57 and 64%, respectively. Overall, increase in the amount of iron sulfate decreased the severity of the disease and enhanced biomass of aerial parts in infected wheat plants.

Antagonistic Activity of Chilean Strains of Pseudomonas protegens Against Fungi Causing Crown and Root Rot of Wheat (Triticum aestivum L.).

Seed treatments with antagonistic bacteria could reduce the severity of crown and root rot diseases in wheat crops. The objective of this study was to evaluate the potential antagonistic activity of a bacterial consortium of three Chilean strains of Pseudomonas protegens against the wheat crown and root rot pathogens Gaeumannomyces graminis var. tritici, Rhizoctonia cerealis, and Fusarium culmorum. Two field experiments were carried out on artificially infested soil during two consecutive seasons (2016–2017 and 2017–2018) in an Andisol soil of southern Chile. Control treatments (not inoculated with fungi) were also included. Each treatment included a seed treatment of spring wheat cv. Pantera-INIA with and without the bacterial consortium. Both phytosanitary damage (incidence and severity) and agronomic components were evaluated. Bacterial populations with the phlD+ gene in the wheat plant rhizosphere during anthesis state (Z6) were also quantified. In both seasons, infection severity decreased by an average of 16.8% in seeds treated with P. protegens consortium, while yield components such as spikes m−1 and number of grains per spike increased. The use of antagonistic bacteria resulted in a total yield increase only during the first experimental season (P < 0.05). In general, accumulated rainfall influenced the antagonistic effect of the consortium of P. protegens strains, accounting for the differences observed between the two seasons. The results suggest that this P. protegens consortium applied on seeds can promote plant growth and protect wheat crops against crown and root rot pathogens in Southern Chile under field conditions.

Novel Fungicide 4-Chlorocinnamaldehyde Thiosemicarbazide (PMDD) Inhibits Laccase and Controls the Causal Agent of Take-All Disease in Wheat, Gaeumannomyces graminis var. tritici.

Our aim was to investigate the bioactivity and mode of action of a novel fungicide 4-chlorocinnamaldehyde thiosemicarbazide (PMDD). As a result of its efficacy against various plant pathogens, its protective fungicidal activity, and systemic transport after root treatment, PMDD could be a promising fungicide to control wheat root diseases. In a field assay, PMDD showed good control efficacy on wheat take-all disease. A biochemical study indicated that PMDD acts as a laccase inhibitor, a to date unique mode of fungicidal action. Moreover, a total of seven stable PMDD-resistant Gaeumannomyces graminis var. tritici (Ggt) mutants were generated and demonstrated no cross-resistance with any commercial fungicides used for take-all disease control, and the gene expression profile further confirmed that laccase could be the target of PMDD. Taken together, we conclude that PMDD is a laccase inhibitor and could be used on wheat to control take-all diseases. The current study could strongly benefit the registration and application of PMDD.

Pseudomonas synxantha 2-79 Transformed with Pyrrolnitrin Biosynthesis Genes Has Improved Biocontrol Activity Against Soilborne Pathogens of Wheat and Canola.

A four-gene operon (prnABCD) from Pseudomonas protegens Pf-5 encoding the biosynthesis of the antibiotic pyrronitrin was introduced into P. synxantha (formerly P. fluorescens) 2-79, an aggressive root colonizer of both dryland and irrigated wheat roots that naturally produces the antibiotic phenazine-1-carboxylic acid and suppresses both take-all and Rhizoctonia root rot of wheat. Recombinant strains ZHW15 and ZHW25 produced both antibiotics and maintained population sizes in the rhizosphere of wheat that were comparable to those of strain 2-79. The recombinant strains inhibited in vitro the wheat pathogens Rhizoctonia solani anastomosis group 8 (AG-8) and AG-2-1, Gaeumannomyces graminis var. tritici, Sclerotinia sclerotiorum, Fusarium culmorum, and F. pseudograminearum significantly more than did strain 2-79. Both the wild-type and recombinant strains were equally inhibitory of Pythium ultimum. When applied as a seed treatment, the recombinant strains suppressed take-all, Rhizoctonia root rot of wheat, and Rhizoctonia root and stem rot of canola significantly better than did wild-type strain 2-79.

Effects of sophorolipids on fungal and oomycete pathogens in relation to pH solubility.

The objective of this study was to determine the effects of sophorolipids on several fungal and oomycete plant pathogens and the relationship between sophorolipids at different pH and antimicrobial activities. Sophorolipids had different solubility at different pH with a dramatic increase in solubility when pH was 6 or higher. Inhibition of mycelial growth of Phytophthora infestans by sophorolipids was affected by pH values, showing that when the pH value was higher, the inhibition rate was lower. Sophorolipids inhibited spore germination and mycelial growth of several fungal and oomycete pathogens in vitro including Fusarium sp., F. oxysporum, F. concentricum, Pythium ultimum, Pyricularia oryzae, Rhizoctorzia solani, Alternaria kikuchiana, Gaeumannomyces graminis var. tritici and P. infestans and caused morphological changes in hyphae by microscope observation. Sophorolipids reduced β-1,3-glucanase activity in mycelia of P. infestans. In greenhouse studies, foliar application of sophorolipids at 3mgml-1 reduced severity of late blight of potato caused by P. infestans significantly. Sophorolipids influenced spore germination and hyphal tip growth of several plant pathogens and pH solubility of sophorolipids had an effect on their efficacy. Application of sophorolipids reduced late blight disease on potato under greenhouse conditions. The findings indicated that sophorolipids have the potential to be developed as a convenient and easy-to-use formulation for managing plant diseases.

The Highly Conserved Barley Powdery Mildew Effector BEC1019 Confers Susceptibility to Biotrophic and Necrotrophic Pathogens in Wheat.

Effector proteins secreted by plant pathogens play important roles in promoting colonization. Blumeria effector candidate (BEC) 1019, a highly conserved metalloprotease of Blumeria graminis f. sp. hordei (Bgh), is essential for fungal haustorium formation, and silencing BEC1019 significantly reduces Bgh virulence. In this study, we found that BEC1019 homologs in B. graminis f. sp. tritici (Bgt) and Gaeumannomyces graminis var. tritici (Ggt) have complete sequence identity with those in Bgh, prompting us to investigate their functions. Transcript levels of BEC1019 were abundantly induced concomitant with haustorium formation in Bgt and necrosis development in Ggt-infected plants. BEC1019 overexpression considerably increased wheat susceptibility to Bgt and Ggt, whereas silencing this gene using host-induced gene silencing significantly enhanced wheat resistance to Bgt and Ggt, which was associated with hydrogen peroxide accumulation, cell death, and pathogenesis-related gene expression. Additionally, we found that the full and partial sequences of BEC1019 can trigger cell death in Nicotiana benthamiana leaves. These results indicate that Bgt and Ggt can utilize BEC1019 as a virulence effector to promote plant colonization, and thus these genes represent promising new targets in breeding wheat cultivars with broad-spectrum resistance.

A Comparative Transcriptomic and Proteomic Analysis of Hexaploid Wheat's Responses to Colonization by Bacillus velezensis and Gaeumannomyces graminis, Both Separately and Combined.

Tritrophic interactions involving a biocontrol agent, a pathogen, and a plant have been analyzed predominantly from the perspective of the biocontrol agent. To explore the adaptive strategies of wheat in response to beneficial, pathogenic, and combined microorganisms, we performed the first comprehensive transcriptomic, proteomic, and biochemical analysis in wheat roots after exposure to Bacillus velezensis CC09, Gaeumannomyces graminis var. tritici, and their combined colonization, respectively. The transcriptional or translational programming of wheat roots inoculated with beneficial B. velezensis showed mild alterations compared with that of pathogenic G. graminis var. tritici. However, the combination of B. velezensis and G. graminis var. tritici activated a larger transcriptional or translational program than for each single microorganism, although the gene expression pattern was similar to that of individual infection by G. graminis var. tritici, suggesting a prioritization of defense against G. graminis var. tritici infection. Surprisingly, pathogen-associated molecular pattern-triggered immunity and effector-triggered immunity made wheat pretreated with B. velezensis more sensitive to subsequent G. graminis var. tritici infection. Additionally, B. velezensis triggered a salicylic acid (SA)-dependent mode of induced systemic resistance that resembles pathogen-induced systemic acquired resistance. Wheat plants mainly depend on SA-mediated resistance, and not that mediated by jasmonic acid (JA), against the necrotrophic pathogen G. graminis var. tritici. Moreover, SA-JA interactions resulted in antagonistic effects regardless of the type of microorganisms in wheat. Further enhancement of SA-dependent defense responses such as lignification to the combined infection was shown to reduce the level of induced JA-dependent defense against subsequent infection with G. graminis var. tritici. Altogether, our results demonstrate how the hexaploid monocot wheat responds to beneficial or pathogenic microorganisms and prolongs the onset of take-all disease through modulation of cell reprogramming and signaling events.

Comparative Transcriptome Profiling of Gaeumannomyces graminis var. tritici in Wheat Roots in the Absence and Presence of Biocontrol Bacillus velezensis CC09.

This study aimed to explore potential biocontrol mechanisms involved in the interference of antagonistic bacteria with fungal pathogenicity in planta. To do this, we conducted a comparative transcriptomic analysis of the “take-all” pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) by examining Ggt-infected wheat roots in the presence or absence of the biocontrol agent Bacillus velezensis CC09 (Bv) compared with Ggt grown on potato dextrose agar (PDA) plates. A total of 4,134 differentially expressed genes (DEGs) were identified in Ggt-infected wheat roots, while 2,011 DEGs were detected in Bv+Ggt-infected roots, relative to the Ggt grown on PDA plates. Moreover, 31 DEGs were identified between wheat roots, respectively infected with Ggt and Bv+Ggt, consisting of 29 downregulated genes coding for potential Ggt pathogenicity factors – e.g., para-nitrobenzyl esterase, cutinase 1 and catalase-3, and two upregulated genes coding for tyrosinase and a hypothetical protein in the Bv+Ggt-infected roots when compared with the Ggt-infected roots. In particular, the expression of one gene, encoding the ABA3 involved in the production of Ggt’s hormone abscisic acid, was 4.11-fold lower in Ggt-infected roots with Bv than without Bv. This is the first experimental study to analyze the activity of Ggt transcriptomes in wheat roots exposed or not to a biocontrol bacterium. Our results therefore suggest the presence of Bv directly and/or indirectly impairs the pathogenicity of Ggt in wheat roots through complex regulatory mechanisms, such as hyphopodia formation, cell wall hydrolase, and expression of a papain inhibitor, among others, all which merit further investigation.

Integration between Pseudomonas protegens strains and fluquinconazole for the control of take-all in wheat

Mixing fungicides with bacteria that produce antimicrobial compounds is an integrated management tool that can reduce root rot pathogens such as Gaeumannomyces graminis var. tritici (Ggt), which causes take-all disease in wheat. This study analyzed the effect of wheat seed treatments on the survival of two strains of Pseudomonas protegens when applied to the seeds alone or integrated with a fungicide a.i. fluquinconazole. Through three field experiments, under wheat monoculture and rotation, inoculated with Ggt, the effect of the bacterial combination with the fungicide seed treatment on yield components was analyzed, and above ground symptoms and root rot severity of take-all disease were determined. In addition, bacterial populations were quantified using PCR and qPCR with phlD + -specific primers in wheat roots during tillering and anthesis throughout two seasons. P. protegens did not show marked effects on the evaluated parameters. However, when combined with fungicide, they increased yield, grain quality, test weight parameters, biomass, and plant height at early stages of plant development; they also decreased the severity of the disease. Initial populations of the bacteria were not affected when seeds were co-treated with fluquinconazole, which was related to an early root colonization and protection in initial stages of development to stabilize their population during anthesis. Results suggest that the integration of P. protegens strains with fluquinconazole is an alternative management strategy for reducing take-all disease in southern Chile.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat.

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. TUBβ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of TUBβ expression. The expression profile of ExoPG assessed using TUBβ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

Organic matter input influences incidence of root rot caused by Rhizoctonia solani AG8 and microorganisms associated with plant root disease suppression in three Australian agricultural soils

Soil-borne plant root disease caused by Rhizoctonia solani AG8 is prevalent in cereal farming systems worldwide, particularly in semiarid agricultural regions. A controlled environment study was undertaken using three Australian soils to test the hypothesis that OM input from crop roots and residues decreases infection by Rhizoctonia root rot via biologically mediated disease suppression. The specific aim was to determine the relative effect of two different OM inputs (wheat stubble or roots) on (a) abundance (DNA) of the pathogen R. solani AG8 and soil organisms putatively associated with disease suppression, and (b) incidence of Rhizoctonia root rot infection of wheat seedlings (% root infected). An increase in microbial biomass carbon (C) following OM amendment indicated a potential for enhanced general biological disease suppression in all soils. OM inputs also increased the population size (DNA) of certain bacteria and fungi putatively associated with specific suppression for Rhizoctonia root rot, suggesting a C resource-mediated change in microbial functions related to disease suppression. There were no significant changes to measured pathogens with stubble addition. However, OM inputs via root residues and rhizodeposits from living roots increased the populations of R. solani AG8 and Gaeumannomyces graminis var. tritici so that in subsequently planted wheat there was greater incidence of root disease infection and reduced plant shoot and root DM compared with that following OM input as stubble. Differences between soils in terms of plant and soil organism responses to each OM input suggest that abiotic factors modify the development of biological disease suppression and the expression of the disease.

Basidiomycetes fungi as biocontrol agents against take-all disease of wheat

Take-all caused by Gaeumannomyces graminis var. tritici (Ggt) is one of the most important root and crown diseases of wheat. In this research, four basidiomycetes isolates as fungal endophytes were isolated from healthy cereal plants and were identified as Coprinopsis urticicola M2, Rhizoctonia endophytica M9, Rhizoctonia zeae M32 and Fomes fomentarius M40 based on phenotypic characteristics and molecular phylogenetic analysis of the ITS rDNA sequences (ITS1-5.8S rDNA-ITS2). Then, their effects as potential biological control agents (BCAs) against Ggt were tested in vitro and greenhouse conditions. All isolates had inhibitory effects on the development of Ggt growth in vitro by dual-culture method with inhibition percentage ranging from 39.1% to 54.8%. Volatile antifungal effects were observed for M32 isolate with 23.7% inhibition, followed by the isolate M2 (18.7%). However, culture filtrates of M2 isolate showed maximum inhibition (29.8%) against Ggt. Only two of the four isolates (M2 and M32) had significant effects on controlling take-all in greenhouse conditions. M2 isolate produced IAA and gibberellin, which had the most effect on increasing of growth parameters of wheat plant in greenhouse conditions. M2 isolate was able to colonize the root cortex of wheat seedlings more rapidly compared to pathogen, as assessed by both cultural and microscopic methods. Therefore, its mode of action may depend on the internal root colonization of wheat by C. urticicola M2 and competition for space and nutrient with the Ggt. This is the first research which shows that the C. urticicola M2 could colonize the wheat roots and has great potential to be used as effective biocontrol agent of take-all. Additionally, it can improve growth parameters in wheat under greenhouse conditions. Therefore, the results of this research indicated that C. urticicola M2, due to its ability to colonize the root and crown internal tissues, may provide an advantage as biocontrol agents to be used in the field conditions.

Microbial Community Composition in Take-All Suppressive Soils.

Gaeumannomyces graminis var. tritici (Ggt) is the main soilborne factor that affects wheat production around the world. Recently we reported the occurrence of six suppressive soils in monoculture areas from indigenous “Mapuche” communities, and evidenced that the suppression relied on the biotic component of those soils. Here, we compare the rhizosphere and endosphere microbial community structure (total bacteria, actinomycetes, total fungi, and ascomycetes) of wheat plants grown in suppressive and conducive soils. Our results suggested that Ggt suppression could be mediated mostly by bacterial endophytes, rather than rhizosphere microorganisms, since the community structure was similar in all suppressive soils as compared with conducive. Interestingly, we found that despite the lower incidence of take-all disease in suppressive soils, the Ggt concentration in roots was not significantly reduced in all suppressive soils compared to those growing in conducive soil. Therefore, the disease suppression is not always related to a reduction of the pathogen biomass. Furthermore, we isolated endophytic bacteria from wheat roots growing in suppressive soils. Among them we identified Serratia spp. and Enterobacter spp. able to inhibit Ggt growth in vitro. Since the disease, but not always pathogen amount, was reduced in the suppressive soils, we propose that take all disease suppressiveness is not only related to direct antagonism to the pathogen.

Construction of a proteome reference map and response of Gaeumannomyces graminis var. tritici to 2,4-diacetylphloroglucinol

Take-all disease, caused by Gaeumannomyces graminis var. tritici (Ggt), is one of the most serious root diseases in wheat production. In this study, a proteomic platform based on 2-dimensional gel electrophoresis (2-DE) and Matrix-Assisted Laser Desorption/Ionization Time of Flight Tandem Mass Spectrometry (MALDI-TOF/TOF MS) was used to construct the first proteome database reference map of G. graminis var. tritici and to identify the response of the pathogen to 2,4-diacetylphloroglucinol (DAPG), which is a natural antibiotic produced by antagonistic Pseudomonas spp. in take-all suppressive soils. For mapping, a total of 240 spots was identified that represented 209 different proteins. The most abundant biological function categories in the Ggt proteome were related to carbohydrate metabolism (21%), amino acid metabolism (15%), protein folding and degradation (12%), translation (11%), and stress response (10%). In total, 51 Ggt proteins were affected by DAPG treatment. Based on gene ontology, carbohydrate metabolism, amino acid metabolism, stress response, and protein folding and degradation proteins were the ones most modulated by DAPG treatment. This study provides the first extensive proteomic reference map constructed for Ggt and represents the first time that the response of Ggt to DAPG has been characterized at the proteomic level.