Showing papers on "Trichoderma harzianum published in 2015"

Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains

Abstract: Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture 'T. harzianum T22' was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1-α (TEF1) is needed to identify species in this complex.

Short title: Trichoderma harzianum systematics Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains

Abstract: Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture ‘T. harzianum T22’ was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1(TEF1) is needed to identify species in this complex.

Solubilisation of Phosphate and Micronutrients by Trichoderma harzianum and Its Relationship with the Promotion of Tomato Plant Growth.

Abstract: Trichoderma harzianum strain SQR-T037 is a biocontrol agent that has been shown to enhance the uptake of nutrients (macro- and microelements) by plants in fields The objective of this study was to investigate the contribution of SQR-T037 to P and microelement (Fe, Mn, Cu and Zn) nutrition in tomato plants grown in soil and in hydroponic conditions Inoculation with SQR-T037 significantly improved the biomass and nutrient uptake of tomato seedlings grown in a nutrient-limiting soil So we investigated the capability of SQR-T037 to solubilise sparingly soluble minerals in vitro via four known mechanisms: acidification by organic acids, chelation by siderophores, redox by ferric reductase and hydrolysis by phytase SQR-T037 was able to solubilise phytate, Fe2O3, CuO, and metallic Zn but not Ca3(PO4)2 or MnO2 Organic acids, including lactic acid, citric acid, tartaric acid and succinic acid, were detected by HPLC and LC/MS in two Trichoderma cultures Additionally, we inoculated tomato seedlings with SQR-T037 using a hydroponic system with specific nutrient deficiencies (ie, nutrient solutions deficient in P, Fe, Cu or Zn and supplemented with their corresponding solid minerals) to better study the effects of Trichoderma inoculation on plant growth and nutrition Inoculated seedlings grown in Cu-deficient hydroponic conditions exhibited increases in dry plant biomass (92%) and Cu uptake (42%) relative to control plants However, we did not observe a significant effect on seedling biomass in plants grown in the Fe- and Zn-deficient hydroponic conditions; by contrast, the biomass decreased by 82% in the P-deficient hydroponic condition Thus, we demonstrated that Trichoderma SQR-T037 competed for P (phytate) and Zn with tomato seedlings by suppressing root development, releasing phytase and/or chelating minerals The results of this study suggest that the induction of increased or suppressed plant growth occurs through the direct effect of T harzianum on root development, in combination with indirect mechanisms, such as mineral solubilisation (including solubilisation via acidification, redox, chelation and hydrolysis)

Influence of Rhizoctonia solani and Trichoderma spp. in growth of bean (Phaseolus vulgaris L.) and in the induction of plant defense-related genes

Abstract: Many Trichoderma species are well-known for their ability to promote plant growth and defense. We study how the interaction of bean plants with R. solani and/or Trichoderma affect the plants growth and the level of expression of defense-related genes. Trichoderma isolates were evaluated in vitro for their potential to antagonize R. solani. Bioassays were performed in climatic chambers and development of the plants was evaluated. The effect of Trichoderma treatment and/or R. solani infection on the expression of bean defense-related genes was analyzed by real-time PCR and the production of ergosterol and squalene was quantified. In vitro growth inhibition of R. solani was between 86 and 58%. In in vivo assays, the bean plants treated with Trichoderma harzianum T019 always had an increased size respect to control and the plants treated with this isolate did not decrease their size in presence of R. solani. The interaction of plants with R. solani and/or Trichoderma affects the level of expression of seven defense-related genes. Squalene and ergosterol production differences were found among the Trichoderma isolates, T019 showing the highest values for both compounds. T. harzianum T019 shows a positive effect on the level of resistance of bean plants to R. solani. This strain induces the expression of plant defense-related genes and produces a higher level of ergosterol, indicating its ability to grow at a higher rate in the soil, which would explain its positive effects on plant growth and defense in the presence of the pathogen.

Colonization of Trichoderma harzianum strain SQR-T037 on tomato roots and its relationship to plant growth, nutrient availability and soil microflora

Abstract: This research was to study the capacity of Trichoderma harzianum strain SQR-T037 to colonize tomato roots and how this strain and its bio-organic fertilizer stimulate plant growth and increase yields under field conditions. Field trials were conducted with a reduced application of chemical fertilizer (75 % of the recommended application) plus Trichoderma-enriched bio-organic fertilizer (BF) or organic fertilizer (OF) or Trichoderma spore suspension (SS), while 100 % of the recommended chemical fertilizer (CF) was used as control. Trichoderma SQR-T037 could efficiently colonize tomato roots and soils based on the reverse transcription-quantitative PCR analysis, and significantly stimulate biomass accumulation at an early stage. The 75 % rates of chemical fertilizer coupled with bio-organic fertilizer (BF) produced tomato yields equivalent to those obtained using the 100 % rates of chemical fertilizer (CF), while inoculation with the Trichoderma alone (SS) or supplement with organic fertilizer alone (OF) would cause 11 and 13 % decreases in yield over the control (CF). The efficacy of BF for maintaining a stable tomato yield may be due to the enhanced soil nutrients availability and the increased abundance of soil microflora, including bacteria, fungi, actinomycetes and Trichoderma communities, which had a positive linear correlation in most of the cases revealed by the Pearson correlation analysis. The results of this study imply that the reason by which the bio-organic fertilizer promotes plant growth and increases the tomato yields can be attributed to the enhancement of the rhizospheric microflora, which promotes nutrient activation. Therefore, T. harzianum could be employed in a combination with composts. In this way, the application rate of chemical fertilizers can be practically decreased by 25 % to obtain maximum benefits.

Biodegradation of polycyclic aromatic hydrocarbons by Trichoderma species: a mini review

Abstract: Fungi belonging to Trichoderma genus are ascomycetes found in soils worldwide. Trichoderma has been studied in relation to diverse biotechnological applications and are known as successful colonizers of their common habitats. Members of this genus have been well described as effective biocontrol organisms through the production of secondary metabolites with potential applications as new antibiotics. Even though members of Trichoderma are commonly used for the commercial production of lytic enzymes, as a biological control agent, and also in the food industry, their use in xenobiotic biodegradation is limited. Trichoderma stands out as a genus with a great range of substrate utilization, a high production of antimicrobial compounds, and its ability for environmental opportunism. In this review, we focused on the recent advances in the research of Trichoderma species as potent and efficient aromatic hydrocarbon-degrading organisms, as well as aimed to provide insight into its potential role in the bioremediation of soils contaminated with heavy hydrocarbons. Several Trichoderma species are associated with the ability to metabolize a variety of both high and low molecular weight polycyclic aromatic hydrocarbons (PAHs) such as naphthalene, phenanthrene, chrysene, pyrene, and benzo[a]pyrene. PAH-degrading species include Trichoderma hamatum, Trichoderma harzianum, Trichoderma reesei, Trichoderma koningii, Trichoderma viride, Trichoderma virens, and Trichoderma asperellum using alternate enzyme systems commonly seen in other organisms, such as multicooper laccases, peroxidases, and ring-cleavage dioxygenases. Within these species, T. asperellum stands out as a versatile organism with remarkable degrading abilities, high tolerance, and a remarkable potential to be used as a remediation agent in polluted soils.

Seed biopriming with drought tolerant isolates of Trichoderma harzianum promote growth and drought tolerance in Triticum aestivum

Abstract: Green house study was aimed to investigate the effect of seed biopriming with drought tolerant isolates of Trichoderma harzianum, viz. Th 56, 69, 75, 82 and 89 on growth of wheat under drought stress and to explore the mechanism underlying plant water stress resilience in response to Trichoderma inoculation. Measurements of relative water content, osmotic potential, osmotic adjustment, leaf gas exchange, chlorophyll fluorescence and membrane stability index were performed. In addition, analysis of the phenolics, proline, lipid peroxidation and measurements of phenylalanine ammonia-lyase activity were carried out. Seed biopriming enhanced drought tolerance of wheat as drought induced changes like stomatal conductance, net photosynthesis and chlorophyll fluorescence were delayed. Drought stress from 4 to 13 days of withholding water induced an increase in the concentration of stress induced metabolites in leaves, while Trichoderma colonisation caused decrease in proline, malondialdehyde (MDA) and hydrogen peroxide (H2O2), and an increase in total phenolics. A common factor that negatively affects plants under drought stress conditions is accumulation of toxic reactive oxygen species (ROS), and we tested the hypothesis that seed biopriming reduced damages resulting from accumulation of ROS in stressed plants. The enhanced redox state of colonised plants could be explained by higher l-phenylalanine ammonia-lyase (PAL) activity in leaves after 13 days of drought stress in Trichoderma treated plants. Similar activity was induced in untreated plants in response to drought stress but to a lower extent in comparison to treated plants. Our results support the hypothesis that seed biopriming in wheat with drought tolerant T. harzianum strains increased root vigour besides performing the process of osmoregulation. It ameliorates drought stress by inducing physiological protection in plants against oxidative damage, due to enhanced capacity to scavenge ROS and increased level of PAL, a mechanism that is expected to augment tolerance to abiotic stresses.

Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi

Abstract: Summary Trichothecenes are phytotoxic sesquiterpenic mycotoxins that can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in HA biosynthesis is the conversion of farnesyl diphosphate to trichodiene (TD), a volatile organic compound (VOC), catalysed by a sesquiterpene synthase encoded by the tri5 gene. Expression of tri5 in the biocontrol strain Trichoderma harzianum CECT 2413 resulted in pro- duction of TD in parallel with a reduction of ergosterol biosynthesis and an unexpected increase in the level of squalene. Transformants expressing tri5 displayed low chitinase activity and induced expression of Botrytis cinerea BOT genes, although their total antagonistic potential against phytopathogenic fungi was not reduced. VOCs released by the tri5- transformant induced expression of tomato defence genes related to salicylic acid (SA), and TD itself strongly induced the expression of SA-responsive genes and reduced the development of lateral roots. Together, these results suggest that TD acts as a signalling VOC in the interactions of Trichoderma with plants and other microorganisms by modulating the perception of this fungus to a given environment.

Biodegradation of pentachlorophenol by marine-derived fungus Trichoderma harzianum CBMAI 1677 isolated from ascidian Didemnun ligulum

Abstract: In this study, the marine-derived fungi Aspergillus sydowii DL6A, Apergillus versicolor DL5A, Cladosporium oxysporum DL5G, Fusarium proliferatum DL11A and Trichoderma harzianum CBMAI 1677 isolated from marine ascidian Didemnun ligulum were evaluated according to their growth in the presence of pentachlorophenol (PCP). The colonies were assessed in 10, 20, 30, 40 and 50 mg L−1 of PCP in a solid culture medium (3% malt). The fungus T. harzianum CBMAI 1677 showed the best growth at 50 mg L−1, which suggests its potential for biodegradation, therefore, this strain was selected for quantitative experiments in 3% malt liquid medium (initial concentration of 20 mg L−1 of PCP) using a validated method. After 7 d of incubation, PCP was not detected and an increasing concentration of pentachloroanisole (PCA) and 2,3,4,6-tetrachloroanisole (2,3,4,6-TeCA) was observed. In a second step, T. harzianum CBMAI 1677 was employed in the biodegradation of PCA and 2,3,4,6-TeCA in a liquid medium. It was observed that both PCA and 2,3,4,6-TeCA were also biodegraded. T. harzianum CBMAI 1677 is a potential strain for bioremediation studies since it was able of biodegrade not only PCP at 20 mg L−1, but also its main metabolites PCA and 2,3,4,6-TeCA.

Draft Whole-Genome Sequence of the Biocontrol Agent Trichoderma harzianum T6776.

Abstract: Trichoderma harzianum T6776 is a promising beneficial isolate whose effects consist of growth promotion, positive response of photosynthetic activity, hormonal signaling, and carbon partitioning in tomato, coupled with biocontrol of plant pathogens. Here, we present the first genome assembly of T6776, providing a useful platform for the scientific community.

Peptaibol, Secondary‐Metabolite, and Hydrophobin Pattern of Commercial Biocontrol Agents Formulated with Species of the Trichoderma harzianum Complex

Abstract: The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics in plant-protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna(®) , Trichosan(®) , Vitalin(®) , Promot(®) WP, and TrichoMax(®) , formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well-established, HPLC/MS-based peptaibiomics approach, it could unequivocally be demonstrated that all of these formulations contained new and recurrent peptaibols, i.e., peptaibiotics carrying an acetylated N-terminus, the C-terminus of which is reduced to a 1,2-amino alcohol. Their chain lengths, including the amino alcohol, were 11, 14, and 18 residues, respectively. Peptaibols were also to be the dominating secondary metabolites in plate cultures of the four strains obtained from four of the Trichoderma- based BCAs, contributing 95% of the UHPLC-UV/VIS peak areas and 99% of the total ion count MS peak area from solid media. Furthermore, species-specific hydrophobins, as well as non-peptaibiotic secondary metabolites, were detected, the latter being known for their antifungal, siderophore, or plant-growth-promoting activities. Notably, none of the isolates produced low-molecular weight mycotoxins.

Beneficial effects of Trichoderma harzianum T-22 in tomato seedlings infected by Cucumber mosaic virus (CMV)

Abstract: The study of the biochemical and molecular mechanisms deriving from the host-pathogen-antagonist interaction is essential to understand the dynamics of infectious processes and can be useful for the development of new strategies to control phytopathogens, particularly viruses, against which chemical treatments have no effect. In this work, we demonstrate the ability of the rhizospheric fungus Trichoderma harzianum strain T-22 (T22) to induce defense responses in tomato (Solanum lycopersicum var. cerasiforme) against Cucumber mosaic virus (CMV, family Bromoviridae, genus Cucumovirus) strain Fny. A granule formulation containing T22 was used for treating the plants before, simultaneously or after the CMV inoculation, in order to study the molecular and biochemical aspects of the interaction between T22 and tomato against the virus. Reactive oxygen species (ROS) and the genes encoding for ROS scavenging enzymes were investigated. Histochemical analysis revealed a different increase in the superoxide anion ( $$ {\text{O}}_{2}^{ \bullet - } $$ ) and hydrogen peroxide (H2O2) content in plants infected by CMV alone or in the presence of T22, confirming the involvement of ROS in plant defense responses. Gene expression analysis suggested a definite improvement in oxidative stress when plants were treated with T22 after inoculation with CMV. In conclusion, our data indicate that Trichoderma harzianum T-22 stimulates the induction of tomato defense responses against CMV, an action that implies the involvement of ROS, pointing towards its use as a treatment rather than as a preventive measure.

Induced resistance to foliar diseases by soil solarization and Trichoderma harzianum

Abstract: The effect of soil solarization and Trichoderma harzianum on induced resistance to grey mould (Botrytis cinerea) and powdery mildew (Podosphaera xanthii) was studied. Plants were grown in soils pretreated by solarization, T. harzianum T39 amendment or both, and then their leaves were inoculated with the pathogens. There was a significant reduction in grey mould in cucumber, strawberry, bean and tomato, and of powdery mildew in cucumber, with a stronger reduction when treatments were combined. Bacillus, pseudomonad and actinobacterial communities in the strawberry rhizosphere were affected by the treatments, as revealed by denaturing gradient gel electrophoresis fingerprinting. In tomato, treatments affected the expression of salicylic acid (SA)-, ethylene (ET)- and jasmonic acid (JA)-responsive genes. With both soil treatments, genes related to SA and ET – PR1a, GluB, CHI9 and Erf1 – were downregulated whereas the JA marker PI2 was upregulated. Following soil treatments and B. cinerea infection, SA-, ET-, and JA-related genes were globally upregulated, except for the LOX genes which were downregulated. Upregulation of the PR genes PR1a, GluB and CHI9 in plants grown in solarized soil revealed a priming effect of this treatment on these genes' expression. The present study demonstrates the capacity of solarization and T. harzianum to systemically induce resistance to foliar diseases in various plants. This may be due to either a direct effect on the plant or an indirect one, via stimulation of beneficial microorganisms in the rhizosphere.

Biological management of Sclerotinia sclerotiorum in pea using plant growth promoting microbial consortium

Abstract: The beneficial plant-microbe interactions play crucial roles in protection against large number of plant pathogens causing disease The present study aims to investigate the growth promoting traits induced by beneficial microbes namely Pseudomonas aeruginosa PJHU15, Trichoderma harzianum TNHU27, and Bacillus subtilis BHHU100 treated singly and in combinations under greenhouse and field conditions to control Sclerotinia sclerotiorum Plants treated with three microbe consortium enhanced plant growth maximally both in the presence and absence of the pathogen Increase in plant length, total biomass, number of leaves, nodules and secondary roots, total chlorophyll and carotenoid content, and yield were recorded in plants treated with microbial consortia Also, a decrease in plant mortality was observed in plants treated with microbial consortia in comparison to untreated control plants challenged with S sclerotiorum Furthermore, the decrease in disease of all the treatments can be associated with differential improvement of growth induced in pea

Evaluation of antifungal activity of silver nanoparticles against some phytopathogenic fungi and Trichoderma harzianum

Abstract: In vitro antifungal activity of silver nanoparticles, at concentrations of 6, 8, 10, 12, 14 and 16 ppm, was studied on five phytopathogenic fungi, and a biocontrol agent. Then effect of silver nanoparticle at 6 ppm (optimum concentration) was evaluated on Macrophomina phaseolina in greenhouse. For in vitro experiment, the fungal isolates were grown on potato dextrose agar medium amended with silver nanoparticles. Radial fungal growth was recorded after 1, 2, 3, 5 and 10 days and mycelial growth inhibition rates were calculated. The most sensitive fungus to nanoparticles was Pythium aphanidermatum, since all tested concentrations showed 100% inhibition during the 10 days of observation.The second most sensitive fungus was Sclerotinia sclerotiorum, since it was able to grow only at concentration of 6 ppm and M. phaseolina was the third in sensitivity since its growth was inhibited in all concentrations after three days. In greenhouse experiments, five treatments including no nanosilver-no pathogen (Negative control), no nanosilver +pathogen (Positive control), 6 ppm nanosilver- no pathogen, 6 ppm nanosilver +pathogen, Carboxin-Thiram (0.15%) +pathogen were compared. Four characters viz shoot and root fresh and dry weights were measured. Based on the greenhouse experimental results, treatments with nanosilver and fungicide gave higher yields than the positive control. The chemical control treatment had the highest measured parameters, while 6 ppm nanosilver +pathogen treatment had the same parameters as negative control. It may therefore be suggested to use nanosilver as a safer alternative to chemical fungicides for control of M. phaseolina.

Imprint Desorption Electrospray Ionization Mass Spectrometry Imaging for Monitoring Secondary Metabolites Production during Antagonistic Interaction of Fungi.

Abstract: Direct analysis of microbial cocultures grown on agar media by desorption electrospray ionization mass spectrometry (DESI-MS) is quite challenging. Due to the high gas pressure upon impact with the surface, the desorption mechanism does not allow direct imaging of soft or irregular surfaces. The divots in the agar, created by the high-pressure gas and spray, dramatically change the geometry of the system decreasing the intensity of the signal. In order to overcome this limitation, an imprinting step, in which the chemicals are initially transferred to flat hard surfaces, was coupled to DESI-MS and applied for the first time to fungal cocultures. Note that fungal cocultures are often disadvantageous in direct imaging mass spectrometry. Agar plates of fungi present a complex topography due to the simultaneous presence of dynamic mycelia and spores. One of the most devastating diseases of cocoa trees is caused by fungal phytopathogen Moniliophthora roreri. Strategies for pest management include the application of endophytic fungi, such as Trichoderma harzianum, that act as biocontrol agents by antagonizing M. roreri. However, the complex chemical communication underlying the basis for this phytopathogen-dependent biocontrol is still unknown. In this study, we investigated the metabolic exchange that takes place during the antagonistic interaction between M. roreri and T. harzianum. Using imprint-DESI-MS imaging we annotated the secondary metabolites released when T. harzianum and M. roreri were cultured in isolation and compared these to those produced after 3 weeks of coculture. We identified and localized four phytopathogen-dependent secondary metabolites, including T39 butenolide, harzianolide, and sorbicillinol. In order to verify the reliability of the imprint-DESI-MS imaging data and evaluate the capability of tape imprints to extract fungal metabolites while maintaining their localization, six representative plugs along the entire M. roreri/T. harzianum coculture plate were removed, weighed, extracted, and analyzed by liquid chromatography-high-resolution mass spectrometry (LC-HRMS). Our results not only provide a better understanding of M. roreri-dependent metabolic induction in T. harzianum, but may seed novel directions for the advancement of phytopathogen-dependent biocontrol, including the generation of optimized Trichoderma strains against M. roreri, new biopesticides, and biofertilizers.

Trichodiene Production in a Trichoderma harzianum erg1-Silenced Strain Provides Evidence of the Importance of the Sterol Biosynthetic Pathway in Inducing Plant Defense-Related Gene Expression

Abstract: Trichoderma species are often used as biocontrol agents against plant-pathogenic fungi. A complex molecular interaction occurs among the biocontrol agent, the antagonistic fungus, and the plant. Terpenes and sterols produced by the biocontrol fungus have been found to affect gene expression in both the antagonistic fungus and the plant. The terpene trichodiene (TD) elicits the expression of genes related to tomato defense and to Botrytis virulence. We show here that TD itself is able to induce the expression of Botrytis genes involved in the synthesis of botrydial (BOT) and also induces terpene gene expression in Trichoderma spp. The terpene ergosterol, in addition to its role as a structural component of the fungal cell membranes, acts as an elicitor of defense response in plants. In the present work, using a transformant of T. harzianum, which is silenced in the erg1 gene and accumulates high levels of squalene, we show that this ergosterol precursor also acts as an important elicitor molecule of tomato defense-related genes and induces Botrytis genes involved in BOT biosynthesis, in both cases, in a concentration-dependent manner. Our data emphasize the importance of a balance of squalene and ergosterol in fungal interactions as well as in the biocontrol activity of Trichoderma spp.

Antifungal Rhizosphere Bacteria Can increase as Response to the Presence of Saprotrophic Fungi

Abstract: Knowledge on the factors that determine the composition of bacterial communities in the vicinity of roots (rhizosphere) is essential to understand plant-soil interactions. Plant species identity, plant growth stage and soil properties have been indicated as major determinants of rhizosphere bacterial community composition. Here we show that the presence of saprotrophic fungi can be an additional factor steering rhizosphere bacterial community composition and functioning. We studied the impact of presence of two common fungal rhizosphere inhabitants (Mucor hiemalis and Trichoderma harzianum) on the composition of cultivable bacterial communities developing in the rhizosphere of Carex arenaria (sand sedge) in sand microcosms. Identification and phenotypic characterization of bacterial isolates revealed clear shifts in the rhizosphere bacterial community composition by the presence of two fungal strains (M. hiemalis BHB1 and T. harzianum PvdG2), whereas another M. hiemalis strain did not show this effect. Presence of both M. hiemalis BHB1 and T. harzianum PvdG2 resulted in a significant increase of chitinolytic and (in vitro) antifungal bacteria. The latter was most pronounced for M. hiemalis BHB1, an isolate from Carex roots, which stimulated the development of the bacterial genera Achromobacter and Stenotrophomonas. In vitro tests showed that these genera were strongly antagonistic against M. hiemalis but also against the plant-pathogenic fungus Rhizoctonia solani. The most likely explanation for fungal-induced shifts in the composition of rhizosphere bacteria is that bacteria are being selected which are successful in competing with fungi for root exudates. Based on the results we propose that measures increasing saprotrophic fungi in agricultural soils should be explored as an alternative approach to enhance natural biocontrol against soil-borne plant-pathogenic fungi, namely by stimulating indigenous antifungal rhizosphere bacteria.

Expression of the β-1,3-glucanase gene bgn13.1 from Trichoderma harzianum in strawberry increases tolerance to crown rot diseases but interferes with plant growth.

Abstract: The expression of antifungal genes from Trichoderma harzianum, mainly chitinases, has been used to confer plant resistance to fungal diseases. However, the biotechnological potential of glucanase genes from Trichoderma has been scarcely assessed. In this research, transgenic strawberry plants expressing the β-1,3-glucanase gene bgn13.1 from T. harzianum, under the control of the CaMV35S promoter, have been generated. After acclimatization, five out of 12 independent lines analysed showed a stunted phenotype when growing in the greenhouse. Moreover, most of the lines displayed a reduced yield due to both a reduction in the number of fruit per plant and a lower fruit size. Several transgenic lines showing higher glucanase activity in leaves than control plants were selected for pathogenicity tests. When inoculated with Colletotrichum acutatum, one of the most important strawberry pathogens, transgenic lines showed lower anthracnose symptoms in leaf and crown than control. In the three lines selected, the percentage of plants showing anthracnose symptoms in crown decreased from 61 % to a mean value of 16.5 %, in control and transgenic lines, respectively. Some transgenic lines also showed an enhanced resistance to Rosellinia necatrix, a soil-borne pathogen causing root and crown rot in strawberry. These results indicate that bgn13.1 from T. harzianum can be used to increase strawberry tolerance to crown rot diseases, although its constitutive expression affects plant growth and fruit yield. Alternative strategies such as the use of tissue specific promoters might avoid the negative effects of bgn13.1 expression in plant performance.

Screening marine-derived endophytic fungi for xylan-degrading enzymes

Abstract: Marine-derived fungi surviving as symptomless endo- phytes in seaweeds and seagrasses were screened for production of xylan-degrading enzymes. Of the eight endophyte isolates obtained from five different sea- grasses and another eight from six different marine algae, half of them exhibited xylanase activity in an agar plate assay. Further examination of these lead candidates using spectrophotometric assays revealed that Trichoderma harzianum, endophytic in the brown alga Sargassum wightii, had the maximum secreted xy- lanase and xylosidase activity. Moreover, this fungus could grow in NaCl-containing media (up to 1.2 M NaCl), and inclusion of 0.26 M NaCl in the media elic- ited a two- and three-fold increase in extracellular xylanase and xylosidase activity respectively. These findings highlight the potential of prospecting marine- derived fungal endophytes to identify novel cell-wall degrading enzymes of value to the biofuel industry.

Exploitation of microbes for enhancing bacoside content and reduction of Meloidogyne incognita infestation in Bacopa monnieri L.

Abstract: Despite the vast exploration of rhizospheric microbial wealth for crop yield enhancement, knowledge about the efficacy of microbial agents as biocontrol weapons against root-knot disease is scarce, especially in medicinal plants, viz., Bacopa monnieri. In the present investigation, rhizospheric microbes, viz., Bacillus megaterium, Glomus intraradices, Trichoderma harzianum ThU, and their combinations were evaluated for the management of Meloidogyne incognita (Kofoid and White) Chitwood and bacoside content enhancement in B. monnieri var CIM-Jagriti. A novel validated method Fourier transform near infrared was used for rapid estimation of total bacoside content. A significant reduction (2.75-fold) in root-knot indices was observed in the combined treatment of B. megaterium and T. harzianum ThU in comparison to untreated control plants. The same treatment also showed significant enhancement (1.40-fold) in total bacoside contents (plant active molecule) content using Fourier transform near-infrared (FT-NIR) method that analyses samples rapidly in an hour without solvent usage and provides ample scope for natural product studies.

Optimization of degradation of winery-derived biomass waste by Ascomycetes

Abstract: BACKGROUND: Recently, winery wastes have been classified as pollutants by the European Union and post-product processing is required to lower their hazards. Individual fungal enzymes have limited capacity so mixed fungal degradation combined with pre-treatment can decrease biomass recalcitrance for more efficient breakdown, overcoming these limitations. RESULTS: Winery biomass degradation by a mixture of Trichoderma harzianum, Aspergillus niger, Penicillium chrysogenum and P. citrinum in submerged fermentation and solid state fermentation (SSF) was evaluated. Higher cellulase and �� -glucosidase activities were observed in SSF and submerged fermentation, respectively. Statistical modelling predicted the fungal percentage ratio of 60:14:4:2 for A. niger: P. chrysogenum: T. harzianum: P citrinum with a substrate:medium ratio of 0.39:1. Under the optimized conditions, cellulase, xylanase and �� -glucosidase activities increased to 78.5, 3544.7 and 250.9 U mL −1 , respectively. Cellulases and xylanases activities increased more than two-fold. Lignin degradation increased from 8% in submerged fermentation (P. chrysogenum) to 17.9% under optimized conditions. Gas chromatography–mass spectrometry (GC-MS) analysis identified 78 significant metabolites, of which stigmasterol, glycerol, maleic acid, xylitol and citric acid were generated by fungal degradation.