Trichoderma reduces scald and protects the photosynthetic apparatus in rice plants

Scald reduces the photosynthetic area, causing yield losses in rice. Changes in gas exchange parameters caused by the pathogen begin before the onset of symptoms. Chemical methods are most commonly applied to control this disease; further research into biological control methods is required. Since Trichoderma asperellum induces plant pathogen defences, increases growth, and improves photosynthetic capability, this study investigated the efficacy of T. asperellum (Ufra T06, UfraT09, Ufra T12, and Ufra T52 (Ta)) in reducing the scald lesion size and the area under the disease progress curve and in minimising the negative effects of scald on gas exchange, chlorophyll a fluorescence, chlorophyll content, and oxidative stress enzyme activity. The experiment was a completely randomised design with five replications and two treatments. Scald was reduced by 62% in plants treated with T. asperellum compared with that in control. There was a 62% increase in the net CO₂ assimilation rate (A) and a drop of 78% in the transpiration rate (E) in plants treated with T. asperellum. The maximum fluorescence (F_m) was 128% higher, and ascorbate peroxidase activity was also higher in plants treated with T. asperellum than in the control. This shows that the use of T. asperellum may be effective in improving the sustainability of the integrated management of rice diseases.     

Aspergillus flavus infection and aflatoxin contamination in sorghum seeds and their biological management

In order to establish the current scenario of aflatoxigenic fungal infection and aflatoxin contamination in sorghum seeds across India, 58 seed samples were collected from different agro-climatic regions. Among these, 67.2% samples were infected with Aspergillus spp. and 28% were found contaminated with aflatoxins ranging from 0.0 to 130?μg?kg^?1. Greenhouse studies revealed no correlation between incidence of Aspergillus flavus and aflatoxin content, and its effect on seed quality parameters. Among the 37 A. flavus strains isolated, six were non-aflatoxigenic when analysed through cultural, TLC and ic-ELISA. Seed treatment with biocontrol agents (antagonistic Rhizobacteria and Trichoderma) suppressed the growth of A. flavus under laboratory and significantly enhanced seed quality variables under greenhouse conditions to a various extent. Field trials with selected biocontrol agents showed that talcum powder formulations of Pseudomonas putida Has-1/c, Bacillus spp. 3/a, Trichoderma asperellum M5 and T. asperellum T2 improved seedling emergence, % nutrient accumulation in plants, increased plant biomass and 1000 seed weight. Seeds harvested from treated plants showed significant increase in seed quality variables under laboratory and greenhouse conditions in comparison with control, but there was no significant difference in A. flavus infection and aflatoxin was completely absent in all treatments.     

Biological Control Agents in Combination with Fertilization or Fumigation to Reduce Sclerotial Viability of Sclerotium rolfsii and Disease of Snap Beans in the Greenhouse

The use of biological control agents in combination with fertilization or fumigation to reduce sclerotial viability of Sclerotium rolfsii and the disease it causes on snap bean was investigated in the greenhouse. The fertilizers ammonium sulphate [(NH₄)₂SO₄], ammonium nitrate (NH₄NO₃), diammonium phosphate [(NH₄)₂HPO₄], or urea applied to soil at a field rate of 135 kg/ha, 15 cm deep of nitrogen (N) (0.09 mg of N/g) or Gliocladium virens (Gl-3) biomass at a rate of 7.5 kg/ha, 15 cm deep (0.05 mg/g) did not reduce the viability of sclerotia of S. rolfsii (Sr-1) when each was applied alone. However, treatment with fertilizer together with the low rate of Gl-3 biomass significantly reduced the sclerotial viability. The treatments that were effective in reducing the viability by more than 75% were the application of (NH₄)₂SO₄ or (NH₄)₂PO₄ and the low rate of Gl-3 biomass. Application of the high rate (0.25 mg/g) of Gl-3 biomass alone only reduced the sclerotial viability by 25%. The addition of any of the fertilizers with the low rate of biomass generally resulted in bean seed germination in the pathogen-infested soil that was higher than that achieved with each individual component. The disease severity (DSI) on beans was appreciable (<3.0) in pathogen-infested soil treated with or without the fertilizer (NH₄)₂SO₄ and in pathogen-infested soil without fertilizer but with a low rate of Gl-3. However, in pathogen-infested soil treated with the fertilizer and the low rate of Gl-3 biomass together, the disease was reduced to a DSI value of less than 1.0. In fumigation studies with metham sodium (Vapam), a dose-response study to investigate the viability of sclerotia of S. rolfsii (Sr-3) indicated that fumigant rates of less than 23.3 μ g/g of soil were sublethal. It was also shown that 5.4 μ g/g of metham sodium was inhibitory to Gl-3 biomass but not to conidia. Consequently, the conidia of isolates Gl-3, Thm-4 of Trichoderma hamatum, and Tv-1 of Trichoderma viride were used together with metham sodium at 17.1 μ g/g of soil. Conidia that were applied to the soil 2 days prior to metham sodium reduced the viability of sclerotia more than each individual component. The results of this study suggest the feasibility of effective disease reduction with an approach utilizing biological control in combination with fertilization or fumigation.     

Genetic and metabolic biodiversity of Trichoderma from Colombia and adjacent neotropic regions

The genus Trichoderma has been studied for production of enzymes and other metabolites, as well as for exploitation as effective biological control agents. The biodiversity of Trichoderma has seen relatively limited study over much of the neotropical region. In the current study we assess the biodiversity of 183 isolates from Mexico, Guatemala, Panama, Ecuador, Peru, Brazil and Colombia, using morphological, metabolic and genetic approaches. A comparatively high diversity of species was found, comprising 29 taxa: Trichoderma asperellum (60 isolates), Trichoderma atroviride (3), Trichoderma brevicompactum (5), Trichoderma crassum (3), Trichoderma erinaceum (3), Trichoderma gamsii (2), Trichoderma hamatum (2), Trichoderma harzianum (49), Trichoderma koningiopsis (6), Trichoderma longibrachiatum (3), Trichoderma ovalisporum (1), Trichoderma pubescens (2), Trichoderma rossicum (4), Trichoderma spirale (1), Trichoderma tomentosum (3), Trichoderma virens (8), Trichoderma viridescens (7) and Hypocrea jecorina (3) (anamorph: Trichoderma reesei), along with 11 currently undescribed species. T. asperellum was the prevalent species and was represented by two distinct genotypes with different metabolic profiles and habitat preferences. The second predominant species, T. harzianum, was represented by three distinct genotypes. The addition of 11 currently undescribed species is evidence of the considerable unresolved biodiversity of Trichoderma in neotropical regions. Sequencing of the internal transcribed spacer regions (ITS) of the ribosomal repeat could not differentiate some species, and taken alone gave several misidentifications in part due to the presence of nonorthologous copies of the ITS in some isolates.     

Characterization of a <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> strain from tomato rhizosphere and its use for integrated management of tomato damping-off

A highly antagonistic Pseudomonas fluorescens strain was isolated from tomato rhizosphere and characterized for its in vitro and in vivo biocontrol potential against Pythium aphanidermatum. The identified Pseudomonas fluorescens strain (PfT-8) was capable of producing high levels of chitinase, β-1,3-glucanase, cellulase, fungitoxic metabolites and siderophores. Seven different carrier formulations including a talc-based powder, lignite-based powder, peat-based powder, lignite + fly ash-based powder, wettable powder, bentonite-paste and polyethylene glycol (PEG) paste were prepared utilizing PfT-8. Shelf life was evaluated for up to 6 months of storage at ambient room temperature (28 °C). Biocontrol efficacy of formulations was studied under greenhouse and field conditions. The formulations were stable up to at least 2 months of storage at ambient room temperature. Among the formulations, peat, lignite, lignite+fly-ash and bentonite paste based formulations maintained higher propagule number than others and also showed greater biocontrol potential. However, propagule number gradually decreased with time. Several organic amendments including farm yard manure (FYM), leaf-compost, poultry manure, press mud, vermi-compost and neem cake were incorporated into soil to study their influence on P. fluorescens colonization in the rhizosphere and on potential disease control. Soil incorporation of organic amendments and specifically poultry manure and FYM, significantly reduced damping-off incidence and also augmented the rhizosphere population of the marked␣P.␣fluorescens strain that was resistant to streptomycin and rifampicin. An integrated␣approach of damping-off management employing dual inoculation of PfT-8 in seed and soil coupled with soil application of organic amendments including poultry manure or␣FYM was evaluated under field conditions. Under these conditions, damping-off incidence substantially reduced by up to 90% and further the healthy plant stand, plant biomass and plant rhizosphere population of P. fluorescens increased significantly.     

Biodegradation of aliphatic hydrocarbon by indigenous fungi isolated from used motor oil contaminated sites

Eighteen indigenous fungal isolates has been successfully isolated from samples of used motor oil, top five centimetres of soil and drainage water contaminated with used motor oil. All of the pure fungal isolates obtained were identified, characterized and subjected to preliminary screening by evaluating the average growth rate of each fungal isolates on minimal media containing 1% (v/v) used motor oil. Trichoderma asperellum strain TUB F-1067 (SA4), Trichoderma asperellum strain Tr48 (SA5), Trichoderma asperellum strain TUB F-756 (SA6), Penicillium species (P1), and Aspergillus species (P9) were further selected for their hydrocarbon biodegradation potential. Among these five fungal isolates selected, P1 strain presented a significant degree of degradation by degrading almost all of the n-alkanes (n-C-₁₅ to n-C-₂₃ range) present in the used motor oil, thus of greater potential in degrading the aliphatic hydrocarbon compounds of used motor oil. The authors would like to certify that the work have not been sent/considered to be published in other journals.     

Production,formulation and antagonistic activity of the biocontrol like-yeast <Emphasis Type="Italic">Aureobasidium pullulans</Emphasis> against <Emphasis Type="Italic">Penicillium expansum</Emphasis>

Aureobasidium pullulans (de Bary) Arnaud (Ach 1-1) was grown in a glucose fed-batch fermentor to 106 g dry wt l⁻¹ in 48 h. The cells were dried in a fluidized bed dryer with a final viability of 62%. After 7 months at 4°C, the viability was 28% of the initial value (= 2.3 × 10¹⁰ c.f.u. g⁻¹ dry matter). A protection level of 89% was achieved with the biomass preparation at 1 × 10⁸ c.f.u. ml⁻¹ after 28 and 7 days for apples stored respectively at 5 and 25°C against Penicillium expansum. Our process is suitable to produce large quantities of the strain Ach 1-1 as biological control agent for apple preservation.     

Potential use of Trichoderma asperellum (Samuels,Liechfeldt et Nirenberg) T8a as a biological control agent against anthracnose in mango (Mangifera indica L.)

Twenty isolates of Trichoderma were obtained from orchards located in three main mango-producing States in Mexico: Chiapas, Oaxaca, and Michoacan, which represent different agronomical management practices and levels of soil fertility. Phylogenetic analysis showed that Trichoderma isolates belong to the following taxa: Hypocrea lixii (10 isolates), Hypocrea jecorina (four isolates), Trichoderma asperellum (three isolates), Trichoderma spirale (two isolates), and Trichoderma brevicompactum (one isolate). The genus Hypocrea is the teleomorph (sexual) stage of the genus Trichoderma, anamorph stage. Seventeen Trichoderma isolates showed at least 67% growth inhibition against the phytopathogenic fungus Colletotrichum gloeosporioides ATCC MYA 456 and three Trichoderma isolates showed complete overgrowth of this pathogen. One member of this group, identified as T. asperellum T8a, was able to control C. gloeosporioides ATCC MYA 456 in vitro and in vivo, as well as five C. gloeosporioides isolates obtained from mango orchards from the State of Oaxaca. Assay of the lytic enzymes involved suggest that cellulases of T. asperellum T8a play a role in biological control against C. gloeosporioides ATCC MYA 456 more than chitinase or glucanase. Thus, native T. asperellum T8a associated with mango trees can be used to enhance mango production, controlling anthracnose through cellulase activity.     

Comparative secretome analysis of <Emphasis Type="Italic">Trichoderma asperellum</Emphasis> S4F8 and <Emphasis Type="Italic">Trichoderma reesei</Emphasis> Rut C30 during solid-state fermentation on sugarcane bagasse

Background

The lignocellulosic enzymes of Trichoderma species have received particular attention with regard to biomass conversion to biofuels, but the production cost of these enzymes remains a significant hurdle for their commercial application. In this study, we quantitatively compared the lignocellulolytic enzyme profile of a newly isolated Trichoderma asperellum S4F8 strain with that of Trichoderma reesei Rut C30, cultured on sugarcane bagasse (SCB) using solid-state fermentation (SSF).

Results

Comparison of the lignocellulolytic enzyme profiles of S4F8 and Rut C30 showed that S4F8 had significantly higher hemicellulase and β-glucosidase enzyme activities. Liquid chromatography tandem mass spectrometry analysis of the two fungal secretomes enabled the detection of 815 proteins in total, with 418 and 397 proteins being specific for S4F8 and Rut C30, respectively, and 174 proteins being common to both strains. In-depth analysis of the associated biological functions and the representation of glycoside hydrolase family members within the two secretomes indicated that the S4F8 secretome contained a higher diversity of main and side chain hemicellulases and β-glucosidases, and an increased abundance of some of these proteins compared with the Rut C30 secretome.

Conclusions

In SCB SSF, T. asperellum S4F8 produced a more complex lignocellulolytic cocktail, with enhanced hemicellulose and cellobiose hydrolysis potential, compared with T. reesei Rut C30. This bodes well for the development of a more cost-effective and efficient lignocellulolytic enzyme cocktail from T. asperellum for lignocellulosic feedstock hydrolysis.

     

Concomitant Induction of Systemic Resistance to Pseudomonas syringae pv. lachrymans in Cucumber by Trichoderma asperellum (T-203) and Accumulation of Phytoalexins

Most studies on the reduction of disease incidence in soil treated with Trichoderma asperellum have focused on microbial interactions rather than on plant responses. This study presents conclusive evidence for the induction of a systemic response against angular leaf spot of cucumber (Pseudomonas syringae pv. lachrymans) following application of T. asperellum to the root system. To ascertain that T. asperellum was the only microorganism present in the root milieu, plants were grown in an aseptic hydroponic growth system. Disease symptoms were reduced by as much as 80%, corresponding to a reduction of 2 orders of magnitude in bacterial cell densities in leaves of plants pretreated with T. asperellum. As revealed by electron microscopy, bacterial cell proliferation in these plants was halted. The protection afforded by the biocontrol agent was associated with the accumulation of mRNA of two defense genes: the phenylpropanoid pathway gene encoding phenylalanine ammonia lyase (PAL) and the lipoxygenase pathway gene encoding hydroxyperoxide lyase (HPL). This was further supported by the accumulation of secondary metabolites of a phenolic nature that showed an increase of up to sixfold in inhibition capacity of bacterial growth in vitro. The bulk of the antimicrobial activity was found in the acid-hydrolyzed extract containing the phenolics in their aglycone form. High-performance liquid chromatography analysis of phenolic compounds showed a marked change in their profile in the challenged, preelicited plants relative to that in challenged controls. The results suggest that similar to beneficial rhizobacteria, T. asperellum may activate separate metabolic pathways in cucumber that are involved in plant signaling and biosynthesis, eventually leading to the systemic accumulation of phytoalexins.     

Fungal Inoculation and Elevated CO2 Mediate Growth of Lolium Mutiforum and Phytolacca Americana,Metal Uptake,and Metal Bioavailability in Metal-Contaminated Soil: Evidence from DGT Measurement

Fungal inoculation and elevated CO₂ may mediate plant growth and uptake of heavy metals, but little evidence from Diffusive Gradients in Thin-films (DGT) measurement has been obtained to characterize the process. Lolium mutiforum and Phytolacca americana were grown at ambient and elevated CO₂ on naturally Cd and Pb contaminated soils inoculated with and without Trichoderma asperellum strain C3 or Penicillium chrysogenum strain D4, to investigate plant growth, metal uptake, and metal bioavailability responses. Fungal inoculation increased plant biomass and shoot/root Cd and Pb concentrations. Elevated CO₂ significantly increased plants biomass, but decreased Cd and Pb concentrations in shoot/root to various extents, leading to a metal dilution phenomenon. Total Cd and Pb uptake by plants, and DGT-measured Cd and Pb concentrations in rhizosphere soils, were higher in all fungal inoculation and elevated CO₂ treatments than control treatments, with the combined treatments having more influence than either treatment alone. Metal dilution phenomenon occurred because the increase in DGT-measured bioavailable metal pools in plant rhizosphere due to elevated CO₂ was unable to match the increase in requirement for plant uptake of metals due to plant biomass increase.     

Expressed sequence tags-based identification of genes in a biocontrol strain Trichoderma asperellum

Trichoderma asperellum, a filamentous soil fungus, is an effective biocontrol agent against many fungal plant pathogenic species. In the present study, we investigated the biological control properties of the strain T. asperellum T4. T. asperellum fermentation products significantly decreased the ability of Rhizoctonia solani and Sclerotinia sclerotiorum to infect rice and soybean, respectively. To further elucidate the biocontrol mechanisms of T. asperellum at the molecular level, a cDNA library was constructed from its mycelium. In total, 3114 expressed sequence tags (ESTs) were generated, which represented 1,554 unigenes, including 354 contigs and 1,200 singletons. Among these unigenes, 731 represented known genes while 823 were novel genes. Forty-six unigenes potentially involved in biocontrol processes were identified from the EST collection. Among them, the expressions of 16 genes were studied, and 15 genes were highly differentially regulated during confrontation with 2 phytopathogens, suggesting that they play roles in the T. asperellum response to phytopathogens. Our study may provide helpful insight in the mechanism of biocontrol by T. asperellum T4 against plant pathogens.     

The importance of water potential range tolerance as a limiting factor on Trichoderma spp. biocontrol of Sclerotinia sclerotiorum

Biological control of fungal phytopathogens is often more variable in efficacy compared with disease suppression achieved by conventional pesticide use. Matching the environmental range of a potential biocontrol agent with that of the target phytopathogen is necessary if consistent disease suppression is to be achieved under field conditions. Strains of Trichoderma that could parasitise sclerotia of Sclerotinia sclerotiorum had their spore germination and mycelial growth (five strains) and ability to parasitise sclerotia (two strains) tested under a range of water potentials under laboratory conditions. Relative mycelial growth and germination of all strains decreased with decreasing osmotic and matric potentials, with matric potential having a greater impact on growth and germination over the range examined. Trichoderma harzianum LU698 mycelial growth was the least affected by decreasing osmotic potential than the other isolates, and Trichoderma atroviride LU141 growth least affected by decreasing matric potential. The germination of LU698 and LU144 was also generally less affected by decreasing osmotic potential, although generally decreasing matric potential had the greatest affect on the germination of LU698 along with T. atroviride LU132. Soil treatments of LU698 and Trichoderma asperellum LU697 reduced sclerotial viability in all but the lowest soil water potential tested, with LU698 being most effective at ?0.1 and ?0.3 MPa after 28 days and LU697 most effective at ?0.01 and ?1.5 MPa after 28 days. We conclude that differences in the tolerance of potential biocontrol agents to changing water potential is an important experimental factor to consider when assaying biocontrol or making predictions of biocontrol efficacy in the field.     

Diversity of root-endophytic <Emphasis Type="Italic">Trichoderma</Emphasis> from Malaysian Borneo

Trichoderma species form endophytic associations with plant roots and may provide a range of benefits to their hosts. However, few studies have systematically examined the diversity of Trichoderma species associated with plant roots in tropical regions. During the evaluation of Trichoderma isolates for use as biocontrol agents, root samples were collected from more than 58 genera in 35 plant families from a range of habitats in Malaysian Borneo. Trichoderma species were isolated from surface-sterilised roots and identified following analysis of partial translation elongation factor-1α (tef1) sequences. Species present included Trichoderma afroharzianum, Trichoderma asperelloides, Trichoderma asperellum, Trichoderma guizhouense, Trichoderma reesei, Trichoderma strigosum and Trichoderma virens. Trichoderma asperellum/T. asperelloides, Trichoderma harzianum s.l. and T. virens were the most frequently isolated taxa. tef1 sequence data supported the recognition of undescribed species related to the T. harzianum complex. The results suggest that tropical plants may be a useful source of novel root-associated Trichoderma for biotechnological applications.     

Formulation of a Streptomyces Biocontrol Agent for the Suppression of Rhizoctonia Damping-off in Tomato Transplants

Formulations of a Streptomyces biological control agent for Rhizoctonia damping-off in tomato seedlings were developed for the first time from vegetative propagules obtained from actively growing, nonsporulating liquid cultures. Alginate beads, durum flour (starch) granules, and talcum powder formulation of this new actinomycetous antagonist (Streptomyces sp. Di-944) isolated from the rhizosphere of field-grown tomato (Lycopersicon esculentum) suppressed damping-off caused by Rhizoctonia solani in tomato plug transplants (cv. Bonny Best) in a peat-based, soilless potting mix under greenhouse conditions. For formulations, vegetative biomass of Streptomyces sp. Di-944 from 3-day-old liquid fermentation in yeast extract–malt extract–glucose broth was lyophilized and pulverized to obtain fragments of viable vegetative filaments. The pulverized biomass had an initial viable count of 2 × 10⁷colony forming units/g and retained 100% viability for 2 weeks when stored at 4°C. Formulations stored at 4°C had a longer shelf life than those stored at 24°C based on viability at 2-week intervals over a 6-month storage period. In addition, dual culture tests showed declining efficacy for surviving Streptomyces propagules in formulations during this storage period. At 4°C, the powder and granular formulations were found to be the most stable and were shown to be 100% viable after 14 and 10 weeks of storage, respectively. However, at the end of 24 weeks, the number of viable propagules in the powder and granular formulations declined to 1.2 × 10⁵ and 7 × 10³ colony forming units/g, respectively. Alginate beads were the least stable in storage. Even at 4°C, 6.9 × 10⁴ and 7.3 × 10² viable propagules/g formulation were detected at the end of 12 and 24 weeks, respectively. The talcum powder formulation delivered to tomato seeds as a seed-coating was the most effective biocontrol treatment. It suppressed damping-off in 10-day-old tomato transplants by almost 90% compared to 30 and 22% damping-off reduction when alginate beads or starch granules were delivered concomitantly with tomato seeds. Seed-coating with powder formulation of the biocontrol agent was as effective as drench application of the fungicide, oxine benzoate (No-Damp), in controlling Rhizoctonia damping-off and superior to the commercial biocontrol agent, Streptomyces griseoviridis (Mycostop), applied to tomato seeds as seed-coating.     

Control of Root-knot Nematode with Formulations of the Nematode-Trapping FungusArthrobotrys dactyloides

Arthrobotrys dactyloidesgrew readily in shaken flasks containing glucose corn steep powder and 8–10 g dry wt of fungal biomass/liter medium was usually produced in 5–6 days. However, it was difficult to convert this biomass into a viable, granulated product suitable for commercial use in biological control. Formulations prepared using kaolin and vermiculite as carriers and gum arabic as a binder showed poor viability when biomass was harvested from liquid culture, mixed with formulation ingredients, granulated, and then dried to a moisture content of less than 5%. Inclusion of a solid-phase incubation step following granulation and prior to drying (incubation of moist granules for 3 days at 25°C in a sterile plastic bag aerated with sterile air) markedly improved biological activity. When granules produced in this manner were placed on a glass slide in field soil, hyphae proliferated from granules and always produced traps. Seven experiments in soil microcosms showed that formulations which had been subjected to solid phase incubation prior to drying consistently reduced numbers ofMeloidogyne javanicajuveniles by more than 90%. In seven glasshouse experiments in which field soils were treated with granules (10 g/liter) and planted to tomatoes, the number of galls induced by the nematode was reduced by 57–96%.     

Phytoextraction of cadmium and zinc by Salix from soil historically amended with sewage sludge

Short rotation coppice (SRC) such as Salix spp. can be grown as an energy crop and offers some potential for economic and practical phytoextraction of marginally contaminated arable soil. This study tested various soil amendments intended to increase soil metal availability to Salix, investigated the distribution of metal between different tree fractions and assessed the viability of phytoextraction using SRC on arable soils. Several Salix genotypes were grown in field trials over 4 years. Cd and Zn concentrations were generally ranked in the order leaves > bark > wood. Metal concentrations in wood increased towards the top of the willow stems, whereas concentrations in leaves showed the opposite trend. None of the amendments significantly increased uptake of Zn by willow. However, in response to a range of soil HCl treatments, mean Cd concentrations in stems and leaves were 112% and 130% of control values. Data from the current experiment, and previous studies, were combined to develop a predictive model of Cd and Zn stem uptake by Salix. The minimum biological concentration factor (BCF) required to achieve a prescribed soil metal target was also calculated based on typical proportions of bioavailable Cd in sludge-amended soils for a 25-year Salix rotation. The best Salix genotypes investigated achieved less than 20% of the uptake rate required to remove one third of the soil Cd content (equivalent to the average isotopically exchangeable Cd fraction in soils at the study site).     

A new oil-based formulation of Trichoderma asperellum for the biological control of cacao black pod disease caused by Phytophthora megakarya

In African cacao-producing countries, control of cacao black pod disease caused by Phytophthora megakarya is a priority. Introducing biological control agents as part of a P. megakarya control strategy is highly desirable, especially in a perspective of pesticide reduction. Trichoderma species are among the most used biological control agents. In Cameroon, Trichoderma asperellum formulated in wettable powder has produced positive effects against this disease. However, with this type of formulation, shelf-life and persistence of conidia on pods are limited. Our study therefore sought to develop a new T. asperellum formulation that would be more effective and better suited to the conditions of field application by small-scale producers in Cameroon. We selected a soybean oil-based oil dispersion, in which the half-life of the conidia reached 22.5 weeks, versus 5 weeks in aqueous suspension. Tested on detached pods, the formulation completely inhibited the development of the disease. When sprayed in the field on cacao clones highly sensitive to P. megakarya, the formulation resulted in 90% protection of treated pods after 1 week, and 50% after 3.2 weeks. The formulations exercised a measurable effect for up to 7 weeks, versus 2 weeks in the case of an aqueous conidial suspension and 5 weeks for that of a conventional fungicide (Kocide). Trichoderma asperellum formulated in oil dispersion has therefore great potential for the control of cacao black pod disease with less recourse to synthetic fungicides. Moreover, this formulation is well adapted to the types of sprayers used by small-scale cacao producers in Cameroon.     

Enzyme activity of extracellular protein induced in Trichoderma asperellum and T. longibrachiatum by substrates based on Agaricus bisporus and Phymatotrichopsis omnivora

Antagonistic Trichoderma spp. are used throughout the world for the biological control of soil-borne plant diseases. This approach has stimulated an on-going search for more efficient mycoparasitic strains with a high potential for producing extracellular lytic enzymes. This study compares the production of lytic enzymes by native strains of Trichoderma asperellum and Trichoderma longibrachiatum on substrates of differing complexity. The quantity of protein induced by Agaricus bisporus-based medium was higher than that induced by Phymatotrichopsis omnivora-based medium. In P. omnivora medium, T. asperellum exhibited higher chitinolytic and β-1,3-glucanolytic activities than T. longibrachiatum. The enzyme profile was related to the previously reported ability of these strains to inhibit the growth of several soil-borne plant pathogens. NAGase production was similar among the tested indigenous strains of T. longibrachiatum; T479 and T359 produced more endochitinase, T479 produced more glucanase, and T341 and T359 produced more β-1,3-glucanase. The detected variations in glucanase and β-1,3-glucanase activities suggest that the production of these enzymes is strongly influenced by the substrate. Strains T397 and T359 exhibited xylanase activity, which triggers defence mechanisms in plants. Thus, these strains may utilise an additional mechanism of biocontrol.     

Biocontrol of vascular streak dieback (Ceratobasidium theobromae) on cacao (Theobroma cacao) through induced systemic resistance and direct antagonism

Vascular streak dieback (VSD), caused by Ceratobasidium theobromae (P.H.B. Talbot &; Keane), has a considerable impact on cacao (Theobroma cacao L.) production in Southeast Asia. Two biocontrol experiments were set up to explore alternatives to ineffective chemical control of VSD. The effects on VSD development of (i) Trichoderma harzianum, superficially applied to leaves of mature cacao plants, and (ii) an inoculation with fungal (Trichoderma asperellum) and bacterial (Bacillus sp., Pseudomonas sp. and Enterobacter spp.) elicitors of systemic resistance in young cacao plants, were tested. After three months, cacao leaves treated with T. harzianum exhibited a significantly (p?p?Bacillus sp. and Enterobacter spp. After transplanting to the field (8 months after inoculation), both number of VSD-affected leaves per plant and number of VSD-affected leaves per branch per plant were significantly (p?T. asperellum did not show potential as an elicitor of systemic resistance in our experiment. More research is needed on the lasting effect of biocontrol treatments, as well as on their economic and ecological sustainability.