Chemical and physical characteristics of wheat root mucilage influenced by Serendipita indica symbiosis: a comparison among four cultivars.

Although there is evidence to suggest that the endophytic fungus Serendipita indica plays a crucial role in enhancing plant tolerance against biotic/abiotic stressors, less is known about the impacts of this symbiosis association on root mucilage chemical composition and its physical functions. The mucilage of inoculated and non-inoculated seedlings of four wheat cultivars (i.e., Roshan, Ghods, Kavir and Pishtaz) were extracted using an aeroponic method. Total solute concentration (TCm), carbon content (Cmucilage), electrical conductivity (EC), pH, fatty acids, surface tension (σm), and viscosity (ηm) of mucilage were measured. Ghods and Kavir had the highest and lowest root colonization percents, respectively. Saturated fatty acids, including palmitic and stearic acids, were dominant over unsaturated fatty acids in wheat root mucilage. However, their compositions were significantly different among wheat cultivars. S. indica colonization, especially for Ghods, increased the TCm, Cmucilage, and palmitic acid. Moreover, root mucilage of S. indica-inoculated Ghods had lower σm and greater ηm. An increased amount of powerful surfactants like palmitic acid in the mucilage of S. indica inoculated treatments led to lower σm and greater ηm. Such studies provide further support for the idea that plant-released mucilage plays a major role in modifying the physical environment of the rhizosphere. This knowledge toward truly understanding the rhizosphere can be potentially used for improving the rhizosphere soil quality and increasing crop growth and yield.

Mycorrhizal colonization and Streptomyces viridosporus HH1 synergistically up-regulate the polyphenol biosynthesis genes in wheat against stripe rust.

BACKGROUND: Stripe rust is considered one of the most devastating diseases of wheat all over the world, resulting in a high loss in its production. In this study, time-course changes in expression of the polyphenol biosynthesis pathways genes in wheat against stripe rust were investigated. The defense mechanisms triggered by mycorrhizal colonization and/or spraying with Streptomyces viridosporus HH1 against this disease were also investigated. RESULTS: Results obtained revealed that C3H, which is considered the key gene in lignin biosynthesis, was the most expressed gene. Furthermore, most of the chlorogenic acid and flavonoid biosynthesis genes were also overexpressed. Volcano plots of the studied genes reveal that the dual treatment led to a high significant overexpression of 10 out of the 13 studied genes. Heatmap of these genes showed that the most frequent expressed gene in response to all applied treatments along the study period was DFR, the key gene in the biosynthesis of anthocyanidins. Gene co-expression network of the studied genes showed that HQT was the most central gene with respect to the other genes, followed by AN2 and DFR, respectively. Accumulation of different flavonoids and phenolic acids were detected in response to the dual treatment, in particular, cinnamic acid, coumarin, and esculetin, which recorded the highest elevation level recording 1000, 488.23, and 329.5% respectively. Furthermore, results from the greenhouse experiment showed that application of the dual treatment led to an 82.8% reduction in the disease severity, compared with the control treatment. CONCLUSIONS: We can conclude that the biosynthesis of lignin, chlorogenic acid, and flavonoids contributed to the synergistic triggering effect of the dual treatment on wheat resistance to stripe rust.

Role of arthroconidia in biofilm formation by Trichosporon asahii.

BACKGROUND: Trichosporon asahii is the major causative agent of disseminated and deep-seated trichosporonosis. It is capable of forming biofilms on surfaces, leading to medical device-related infection.Trichosporon asahii may be present as yeast form, hyphae and/or arthroconidia; however, the relationship between its biofilm-forming ability and its morphological transition is unclear. OBJECTIVES: We investigated whether the T. asahii morphological transition contributes to its biofilm formation. We also determined the conditions required to induce each of the morphologies. METHODS: Three high- and three low-biofilm-producing strains (HBS and LBS, respectively) were selected using a biofilm formation assay, and the cell surface hydrophobicity of these six strains was measured. For each strain, the morphology was observed and the number of each morphological form (yeast form, hypha and arthroconidium) was counted to calculate the ratio. Finally, the ability of cells each morphological type to adhere to the polystyrene substrate was evaluated. RESULTS: The HBS exhibited abundant arthroconidia and hyphae; in contrast, the LBS produced mainly hyphae with few or no arthroconidia. The production of hyphae was increased by nitrogen-containing medium, and the production of arthroconidia was increased by nitrogen-deficient medium. Cells incubated under nitrogen-deficient conditions showed higher adherence to a polystyrene surface than those incubated in the presence of nitrogen. CONCLUSION: Arthroconidia of T. asahii play a key role in biofilm formation by promoting cellular adhesion.

A Poly(A) Ribonuclease Controls the Cellotriose-Based Interaction between Piriformospora indica and Its Host Arabidopsis.

Piriformospora indica, an endophytic root-colonizing fungus, efficiently promotes plant growth and induces resistance to abiotic stress and biotic diseases. P. indica fungal cell wall extract induces cytoplasmic calcium elevation in host plant roots. Here, we show that cellotriose (CT) is an elicitor-active cell wall moiety released by P. indica into the medium. CT induces a mild defense-like response, including the production of reactive oxygen species, changes in membrane potential, and the expression of genes involved in growth regulation and root development. CT-based cytoplasmic calcium elevation in Arabidopsis (Arabidopsis thaliana) roots does not require the BAK1 coreceptor or the putative Ca2+ channels TPC1, GLR3.3, GLR2.4, and GLR2.5 and operates synergistically with the elicitor chitin. We identified an ethyl methanesulfonate-induced mutant (cytoplasmiccalcium elevation mutant) impaired in the response to CT and various other cellooligomers (n = 2-7), but not to chitooligomers (n = 4-8), in roots. The mutant contains a single nucleotide exchange in the gene encoding a poly(A) ribonuclease (AtPARN; At1g55870) that degrades the poly(A) tails of specific mRNAs. The wild-type PARN cDNA, expressed under the control of a 35S promoter, complements the mutant phenotype. Our identification of cellotriose as a novel chemical mediator casts light on the complex P. indica-plant mutualistic relationship.

Trichoderma species mediated differential tolerance against biotic stress of phytopathogens in Cicer arietinum L.

Trichoderma spp. have been reported to aid in imparting biotic as well as abiotic tolerance to plants. However, there are only few reports unfolding the differential ability of separate species of Trichoderma genera generally exploited for their biocontrol potential in this framework. A study was undertaken to evaluate the biocontrol potential of different Trichoderma species namely T. harzianum, T. asperellum, T. koningiopsis, T. longibrachiatum, and T. aureoviride as identified in the group of indigenous isolates from the agricultural soils of Eastern Uttar Pradesh, India. Their biocontrol potential against three major soilborne phytopathogens, i.e., Sclerotium rolfsii, Sclerotinia sclerotiorum, and Colletotrichum capsici was confirmed by dual culture plate technique. Efficient mycoparasitic ability was further assessed in all the isolates in relation to chitinase, ß-1,3 glucanase, pectinase, lipase, amylase, and cellulase production while equally consistent results were obtained for their probable phosphate solubilization and indole acetic acid (IAA) production abilities. The selected isolates were further subjected to test their ability to promote plant growth, to reduce disease incidence and to tolerate biotic stress in terms of lignification pattern against S. rolfsii in chickpea plants. Among the identified Trichoderma species, excellent results were observed for T. harzianum and T. koningiopsis indicating better biocontrol potential of these species in the group and thus exhibiting perspective for their commercial exploitation.

Xylem defense wood of Norway spruce compromised by the pathogenic white-rot fungus Heterobasidion parviporum shows a prolonged period of selective decay.

Heterobasidion parviporum, a common pathogenic white-rot fungus in managed Norway spruce forests in northern and central Europe, causes extensive decay columns within stem heartwood of the host tree. Infected trees combat the lateral spread of decay by bordering the heartwood with a fungistatic reaction zone characterized by elevated pH and phenol content. To examine the mode of fungal feeding in the reaction zone of mature Norway spruce trees naturally infected by H. parviporum, we conducted spatial profiling of pectin and hemicellulose composition, and established transcript levels of candidate fungal genes encoding enzymes involved in degradation of the different cell wall components of wood. Colonized inner heartwood showed pectin and hemicellulose concentrations similar to those of healthy heartwood, whereas the carbohydrate profiles of compromised reaction zone, irrespective of the age of fungal activity in the tissue, indicated selective fungal utilization of galacturonic acid, arabinose, xylose and mannose. These data show that the rate of wood decay in the reaction zone is slow. While the up-regulation of genes encoding pectinases and hemicellulases preceded that of the endoglucanase gene during an early phase of fungal interaction with xylem defense, the manganese peroxidase gene showed similar transcript levels during different phases of wood colonization. It seems plausible that the reaction zone components of Norway spruce interfere with both lignin degradation and the associated co-hydrolysis of hemicelluloses and pectin, resulting in a prolonged phase of selective decay.

Piriformospora indica symbiosis improves water stress tolerance of rice through regulating stomata behavior and ROS scavenging systems.

Global water shortage seriously threatens rice growth especially in irrigated production areas. Association of plants with beneficial soil microbes is one strategy for plant adaption to environmental stresses. In this study, rice (Oryza sativa L.) plants were colonized by the beneficial root-colonizing endophytic fungus Piriformospora indica (P. indica). We demonstrate that grain yield were higher in P. indica-colonized rice plants compared to the uncolonized plants grown in soil. Moreover, P. indica effect on improving water stress tolerance in rice and its physiological mechanism were investigated in a hydroponic culture system. Polyethylene glycol (PEG) was applied to the culture solution to conduct the water stress condition. Water stress-induced leaf wilting and impairments in photosynthetic efficiency were diminished in P. indica-colonized plants. Furthermore, P. indica colonization promotes stomata closure and increases the leaf surface temperature under water stress. The malondialdehyde level (as an indicator for oxidative stress) was lower and the reduced to oxidized glutathione ratio was higher in P. indica-colonized and PEG-exposed rice plants compared to the uncolonized plants. Furthermore, the activities of the antioxidant enzymes catalase and glutathione reductase were up-regulated in inoculated rice seedlings under water stress. In conclusion, P. indica promotes rice performance under water stress by stomata closure and lower oxidative stress.

Development of fungal mycelia as skin substitutes: effects on wound healing and fibroblast.

In this study, Sacchachitin membrane, prepared from the residue of the fruiting body of Ganoderma tsugae, was estimated for its effects on wound healing and the proliferation and migration of fibroblast cells. Two mirror-image wounds were made on the back of female guinea pigs by dissecting a 1.5 x 1.5 cm2 skin surface of full thickness. Sacchachitin membrane was placed randomly on one of the wounds and gauze or Beschitin on the other. Changes in the wound area were measured and photographed after a predetermined amount of time postoperatively. Histological examination of the wound and surrounding tissue was also performed to reveal any interaction of tissue with the dressing. The results showed that the wound area covered with Sacchachitin membrane was statistically smaller than that covering with gauze on day 10, whereas there was no significant difference in the wound size compared to that with Beschitin. Fibroblast cells from the dermis layer of guinea pigs were used. The number of fibroblast cells were counted on the predetermined days in the culture suspended with or without 0.01% w/v dressing materials. By layering on DMEM plates, the number of fibroblast cells migrating across the center line or outside of the central hole were counted after five days. All the results indicated that both 0.01% w/v of Sacchachitin and chitin significantly enhanced the proliferation and migration of fibroblast cells.

[Biological treatments for contaminated soils: hydrocarbon contamination. Fungal applications in bioremediation treatment]. / Tratamientos biológicos de suelos contaminados; contaminación por hidrocarburos. Aplicaciones de hongos en tratamientos de biorrecuperación.

Bioremediation is a spontaneous or controlled process in which biological, mainly microbiological, methods are used to degrade or transform contaminants to non or less toxic products, reducing the environmental pollution. The most important parameters to define a contaminated site are: biodegradability, contaminant distribution, lixiviation grade, chemical reactivity of the contaminants, soil type and properties, oxygen availability and occurrence of inhibitory substances. Biological treatments of organic contaminations are based on the degradative abilities of the microorganisms. Therefore the knowledge on the physiology and ecology of the biological species or consortia involved as well as the characteristics of the polluted sites are decisive factors to select an adequate biorremediation protocol. Basidiomycetes which cause white rot decay of wood are able to degrade lignin and a variety of environmentally persistent pollutants. Thus, white rot fungi and their enzymes are thought to be useful not only in some industrial process like biopulping and biobleaching but also in bioremediation. This paper provides a review of different aspects of bioremediation technologies and recent advances on ligninolytic metabolism research.

Assessment of wood-inhabiting Basidiomycetes for biokraft pulping of softwood chips.

Wood-inhabiting Basidiomycetes have been screened for various applications in the pulp and paper industry and it is evident that different fungi need to be used to suit the specific requirements of each application. This study assessed the suitability of 278 strains of South African wood-decay fungi for the pre-treatment of softwood chips for kraft pulping. The influence of these fungi on kappa number, yield and strength properties of pulp was evaluated. A number of these strains were more efficient in reducing kappa number than the frequently used strains of Phanerochaete chrysosporium and Ceriporiopsis subvermispora. Six strains of Stereum hirsutum and a strain of an unidentified species were able to reduce the kappa number significantly without a significant influence on the pulp yield. Treatment of wood with two strains of S. hirsutum, one strain of Peniophora sp. and a strain of an unidentified species resulted in paper with improved strength properties.

Straw bio-degradation by acidogenic bacteria and composite fungi.

A composite microbial system, including a strain of Candida tropicalis (W3), a strain of Lactobacillus plantarm(WY3) and three strains of basidiomycete pL104, pL113 and C33, was chosen to degrade corn straw. The final pH was acid owing to the inoculation of acidogenic bacteria, and under this condition the composite fungi system could produce complex enzyme to destroy the compact structure of corn straw. The experimental results showed that the biomass of composite fungi could reach up to maximum when the pH value was 4.5. Through the bio-degradation by combining acidogenic bacteria with the composite fungi system, the cellulose, hemi-cellulose and lignin degradation rates of corn straw powder were 26.36%, 43.30% and 26.96%, respectively. And the gross crude protein content increased 60.41%. This study provided the evidence for the feasibility of developing a composite microbial system with high capability of degrading straw lignocelluloses in order to make reasonable use of straw resource and protect rural eco-environment.

Degradation of azo dye acid orange 7 in a membrane bioreactor by pellets and attached growth of Coriolus versicolour.

The aim of this study was to systematically compare the degradation of azo dye acid orange 7 by spongy pellets and attached biofilm of Coriolus versicolour (NBRC 9791) in a membrane bioreactor (MBR) under non-sterile conditions. Mild stirring (35 rpm) resulted in spherical (φ=0.5 cm), spongy pellets and concomitantly triggered high enzymatic activity of the fungus, allowing for excellent decolouration (>99%) of a synthetic wastewater containing the dye. However, bacterial contamination eventually damaged the fungus pellets, leading to decreased decolouration efficiency. Promotion of attached growth on a plastic support along with formation of spherical spongy pellets allowed maintenance of high enzymatic activity and decolouration/degradation for an extended period. Hydraulic retention time (HRT) could influence the level of enzymatic activity and decolouration; however, even at the shortest HRT (1 day) examined, the MBR could accomplish >95% decolouration.

Direct ethanol production from cellulosic materials by the hypersaline-tolerant white-rot fungus Phlebia sp. MG-60.

White-rot fungus Phlebia sp. MG-60 was identified as a good producer of ethanol from several cellulosic materials containing lignin. When this fungus was cultured with 20 g/L unbleached hardwood kraft pulp (UHKP), 8.4 g/L ethanol was produced after 168 h of incubation giving yields of ethanol of 0.42 g/g UHKP, 71.8% of the theoretical maximum. When this fungus was cultured with waste newspaper, 4.2g/L ethanol was produced after 216 h of incubation giving yields of ethanol of 0.20 g/g newspaper, 51.1% of the theoretical maximum. Glucose, mannose, galactose, fructose and xylose were completely assimilated by Phlebia sp. MG-60 with ethanol yields of 0.44, 0.41, 0.40, 0.41 and 0.33 g/g of sugar respectively. These results indicated that Phlebia sp. MG-60 was a good candidate for bioethanol production from cellulosic materials.

Lignocellulose modifications by brown rot fungi and their effects, as pretreatments, on cellulolysis.

Brown rot fungi Gloeophyllum trabeum and Postia placenta were used to degrade aspen, spruce, or corn stover over 16 weeks. Decayed residues were saccharified using commercial cellulases or brown rot fungal extracts, loaded at equal but low endoglucanase titers. Saccharification was then repeated for high-yield samples using full strength commercial cellulases. Overall, brown rot pretreatments enhanced yields up to threefold when using either cellulase preparation. In the best case, aspen degraded 2 weeks by G. trabeum yielded 72% glucose-from-cellulose, a 51% yield relative to original glucan. A follow-up trial with more frequent harvests showed similar patterns and demonstrated interplay between tissue modifications and saccharification. Hemicellulose and vanillic acid (G6) or vanillin (G4) lignin residues were good predictors of saccharification potential, the latter notable given lignin's potential active role in brown rot. Results show basic relationships over a brown rot time course and lend targets for controlling an applied bioconversion process.

Indications of heightened constitutive or primed host response affecting the lignin pathway transcripts and phenolics in mature Norway spruce clones.

Two mature clones of Norway spruce (Picea abies (L.) Karst.) that have previously been shown to have differential degrees of resistance towards the necrotrophic pathogen Heterobasidion parviporum (Niemelä & Korhonen) were compared with respect to the primed defense expression of transcripts related to biosynthesis of lignin, stilbenes and other phenolic compounds from one year to the next. The host's response to physical wounding and pathogen inoculation was examined in the initial year, whereas indications of heightened basal defense level or primed response, and responses to re-wounding, were examined the following year. The responses of the two clones to wounding and pathogen inoculation, examined in the initial year, differed; the increases in lignin and phenolics were more distinct in response to the pathogen than to wounding alone. The more resistant clone 589 had higher initial lignin concentrations in the cell walls when compared with clone 409, and these remained higher in clone 589 over both years and increased after the treatments. Both clones responded at the transcriptional and chemical levels to wounding; changes were evident both in the initial wounds and when re-wounded the following year. There were distinct differences in the basal transcript levels of the lignin pathway-related genes, phenolics and total lignin levels in healthy tissue from the initial year to the following year indicative of a primed host response or at least altered constitutive level of defense expression.

Response of living tissues of Pinus sylvestris to the saprotrophic biocontrol fungus Phlebiopsis gigantea.

The saprotrophic fungus Phlebiopsis gigantea has been used for several years as a biocontrol agent against the conifer pathogen Heterobasidion annosum. Although the effectiveness of P. gigantea in biocontrol has been shown empirically, the long-term effect on living conifer trees as well as the mechanism underlying its antagonistic activity is still unknown. An additional concern is the potential of P. gigantea to acquire a necrotrophic habit through adaptation to living wood tissues. By using a combination of histochemical, molecular and transcript profiling (454 sequencing), we investigated under in vitro conditions the necrotrophic capability of P. gigantea and induced localized resistance as a mechanism for its biocontrol action. Pinus sylvestris seedlings (10 years old) were challenged on the xylem surface with P. gigantea or H. annosum. Both fungi provoked strong necrotic lesions, but after prolonged incubation, P. gigantea lesions shrank and ceased to expand further. Tree seedlings pre-treated with P. gigantea further restricted H. annosum-induced necrosis and had more lignified cells. The 454 sequencing revealed elevated transcript levels of genes important for lignification, cell death regulation and jasmonic acid signalling. The results suggest that induced localized resistance is a contributory factor for the biocontrol efficacy of P. gigantea, and it has a limited necrotrophic capability compared with H. annosum.

The plant cell wall-decomposing machinery underlies the functional diversity of forest fungi.

Brown rot decay removes cellulose and hemicellulose from wood--residual lignin contributing up to 30% of forest soil carbon--and is derived from an ancestral white rot saprotrophy in which both lignin and cellulose are decomposed. Comparative and functional genomics of the "dry rot" fungus Serpula lacrymans, derived from forest ancestors, demonstrated that the evolution of both ectomycorrhizal biotrophy and brown rot saprotrophy were accompanied by reductions and losses in specific protein families, suggesting adaptation to an intercellular interaction with plant tissue. Transcriptome and proteome analysis also identified differences in wood decomposition in S. lacrymans relative to the brown rot Postia placenta. Furthermore, fungal nutritional mode diversification suggests that the boreal forest biome originated via genetic coevolution of above- and below-ground biota.

Evaluation of white-rot fungi-assisted alkaline/oxidative pretreatment of corn straw undergoing enzymatic hydrolysis by cellulase.

In this study, the effects of biological treatment prior to alkaline/oxidative (A/O) pretreatment using three white-rot fungi (Ganoderma lucidum, Trametes versicolor and Echinodontium taxodii) were evaluated for the enzymatic hydrolysis of corn straw. Among these fungi, Echinodontium taxodii significantly enhanced the efficiency of chemical pretreatment. Subsequent to treatment of corn straw with Echinodontium taxodii for 15 days, the straw was subjected to digestion by 0.0016% NaOH and 3% H2O2 at room temperature for 24 h, which increased the reducing sugar yield by 50.7%. The hydrolysis model and kinetic parameters were determined from time course data collected throughout the hydrolysis. The initial hydrolysis rate, V0, of the corn straw increased by 68.5% compared to A/O pretreatment alone, which resulted from an increase in the initial adsorption. The lignin content of the corn straw decreased more significantly after biological and A/O pretreatment than after A/O pretreatment alone. After 72 h of enzymatic hydrolysis, the adsorbed cellulase decreased by 24.8% (from 3.67 to 2.76 mg ml⁻¹) compared to A/O pretreatment alone. These results indicate that biological treatment improves the desorption of cellulase by enhancing delignification during A/O pretreatment.

Piriformospora indica confers drought tolerance in Chinese cabbage leaves by stimulating antioxidant enzymes, the expression of drought-related genes and the plastid-localized CAS protein.

Piriformospora indica, a root-colonizing endophytic fungus of Sebacinales, promotes plant growth and confers resistance against biotic and abiotic stress. The fungus strongly colonizes the roots of Chinese cabbage, promotes root and shoot growth, and promotes lateral root formation. When colonized plants were exposed to polyethylene glycol to mimic drought stress, the activities of peroxidases, catalases and superoxide dismutases in the leaves were upregulated within 24h. The fungus retarded the drought-induced decline in the photosynthetic efficiency and the degradation of chlorophylls and thylakoid proteins. The expression levels of the drought-related genes DREB2A, CBL1, ANAC072 and RD29A were upregulated in the drought-stressed leaves of colonized plants. Furthermore, the CAS mRNA level for the thylakoid membrane associated Ca(2+)-sensing regulator and the amount of the CAS protein increased. We conclude that antioxidant enzyme activities, drought-related genes and CAS are three crucial targets of P. indica in Chinese cabbage leaves during the establishment of drought tolerance. P. indica-colonized Chinese cabbage provides a good model system to study root-to-shoot communication.