Showing papers on "Trichoderma longibrachiatum published in 2015"

Deep RNA sequencing reveals a high frequency of alternative splicing events in the fungus Trichoderma longibrachiatum

Abstract: Alternative splicing is crucial for proteome diversity and functional complexity in higher organisms. However, the alternative splicing landscape in fungi is still elusive. The transcriptome of the filamentous fungus Trichoderma longibrachiatum was deep sequenced using Illumina Solexa technology. A total of 14305 splice junctions were discovered. Analyses of alternative splicing events revealed that the number of all alternative splicing events (10034), intron retentions (IR, 9369), alternative 5’ splice sites (A5SS, 167), and alternative 3’ splice sites (A3SS, 302) is 7.3, 7.4, 5.1, and 5.9-fold higher, respectively, than those observed in the fungus Aspergillus oryzae using Illumina Solexa technology. This unexpectedly high ratio of alternative splicing suggests that alternative splicing is important to the transcriptome diversity of T. longibrachiatum. Alternatively spliced introns had longer lengths, higher GC contents, and lower splice site scores than constitutive introns. Further analysis demonstrated that the isoform relative frequencies were correlated with the splice site scores of the isoforms. Moreover, comparative transcriptomics determined that most enzymes related to glycolysis and the citrate cycle and glyoxylate cycle as well as a few carbohydrate-active enzymes are transcriptionally regulated. This study, consisting of a comprehensive analysis of the alternative splicing landscape in the filamentous fungus T. longibrachiatum, revealed an unexpectedly high ratio of alternative splicing events and provided new insights into transcriptome diversity in fungi.

Quantifying Synergy, Thermostability, and Targeting of Cellulolytic Enzymes and Cellulosomes with Polymerization-Based Amplification

Abstract: We present a polymerization-based assay for determining the potency of cellulolytic enzyme formulations on pretreated biomass substrates. Our system relies on monitoring the autofluorescence of cellulose and measuring the attenuation of this fluorescent signal as a hydrogel consisting of poly(ethylene glycol) (PEG) polymerizes on top of the cellulose in response to glucose produced during saccharification. The one-pot method we present is label-free, rapid, highly sensitive, and requires only a single pipetting step. Using model enzyme formulations derived from Trichoderma reesei, Trichoderma longibrachiatum, Talaromyces emersonii and recombinant bacterial minicellulosomes from Clostridium thermocellum, we demonstrate the ability to differentiate enzyme performance based on differences in thermostability, cellulose-binding domain targeting, and endo/exoglucanase synergy. On the basis of its ease of use, we expect this cellulase assay platform to be applicable to enzyme screening for improved bioconversion of lignocellulosic biomass.

Enhanced Biodegradation of Endosulfan by Aspergillus and Trichoderma spp. Isolated from an Agricultural Field of Tarai Region of Uttarakhand

Abstract: Two endosulfan degrading fungal isolates (FII and FIII) were recovered from the soil of an agricultural field of Udham Singh Nagar, Uttarakhand. Maximum tolerance limit of both the fungi for endosulfan was 350 ppm in minimal medium. On the basis of morphological and molecular characters, FII and FIII showed 100% homology with Aspergillus oryzae and Trichoderma longibrachiatum respectively. Pattern of degradation of endosulfan in minimal broth was: consortium (FII+FIII) >FII>FIII and percent degradation of # and # isomers of endosulfan by the fungal consortium was 93 and 95 respectively after 15 days. Immobilized fungal consortium performed better than the individual fungus and consortium under same conditions. Consortium performed best in biodegradation and uptake of endosulfan in a pot experiment on maize, where soil was spiked with 100 mg kg−1 endosulfan.

Trichoderma Transformation Methods

Abstract: Trichoderma includes a huge variety of fungal species with an increasing interest from several points of view. Many of their species produce primary or secondary metabolites with importance in pharmaceutical industry or in the biocontrol of significant phytopathogenic fungi (Cardoza et al., Curr Genet 34:50–59, 2005; Sivasithamparam and Ghisalberti, Trichoderma and Gliocladium, London, pp 139–191). Other Trichoderma species produce enzymes with a remarkable industrial importance and even other strains from Trichoderma brevicompactum (Tijerino et al., Fungal Genet Biol 48:285–296, 2011a, Toxins (Basel) 3:1220–1232, 2011b) or Trichoderma longibrachiatum (Alanio et al., Clin Infect Dis 46:e116–118, 2008) have potential pathogenic activity against plants and animals, including human beings, respectively. The practical applications of this genus have resulted in an increasing interest in the development of efficient transformation procedures, in order to select transgenic strains with valuable phenotypes. These procedures should be essential tools to characterize the physiological roles of the increasing number of genes available as a result of the fungal genomic projects (http://genome.jgi-psf.org/programs/fungi) (Grigoriev et al., Nucleic Acids Res 40:D26–32, 2012).

Allelopathic potential of some biocontrol agents for the control of fungal rot of yellow yam (Dioscorea cayenensis Lam)

Abstract: The adverse effect of synthetic pesticides on human health and the natural ecosystem necessitate the need to explore natural mechanisms of disease control in plants. This study evaluated the allelopathic potential of five biocontrol agents: Trichoderma longibrachiatum, Trichoderma asperellum, Bacillus subtilis, Bacillus cereus and Pseudomonas fluorescens in the control of six fungal pathogens associated with tuber rot of Dioscorea cayenensis. Rotten tuber samples were randomly collected across three agro-ecological zones (AEZs): humid rainforest (HF), derived savanna (DS), and southern Guinea savanna (SGS) in Nigeria. Biocontrol agents were isolated from the yam rhizosphere using the serial dilution method; the agar pairing method was used for the in vitro trials. The destructive sampling method was used to evaluate rot control by the antagonists in vivo. Aspergillus niger had the highest incidence of 64.71% across the HF, 52.08% across the DS, and 41.98% across the SGS. B. subtilis had the highest inhibitory zone of 16.7 ± 0.05% when paired with A. niger, 15.4 ± 0.01% with Lasiodiplodia theobromae, 14.0 ± 0.33% with Penicillium oxalicum, 7.1 ± 0.14% when paired with Rhizoctonia solani; 17.1 ± 0.11% with Sclerotium rolfsii, and 10.3 ± 0.94% with Fusarium oxysporum. All biocontrol agents significantly (P = 0.05) reduced rot development of the test pathogens relative to the control in the in vivo experiment. The establishment of a distinct zone of inhibition, especially by the bacterial antagonists attests to the fact that they produced allelochemical substances. Therefore, further research is recommended to evaluate the biochemical composition of these microbial metabolites, their level of toxicity, and fate in the environment. Key words: Allelopathic potential, biocontrol agents, tubers, allelochemicals

Improved production of -galactosidase and -fructofuranosidase by fungi using alternative carbon sources

Abstract: The influence of alternative carbon sources as inducers of i¢-galactosidase and i¢-fructofuranosidase by filamentous fungi (that are, Aspergillus aculeatus, Chrysonilia sitophila, Gliocladium virens, Aspergillus fumigatus and Trichoderma longibrachiatum), recently isolated from Brazil’s Atlantic Forest biome has been investigated. The greatest levels of intracellular β-galactosidase activity were obtained using orange peel waste (56.31 U/mL) with A. aculeatus, rice straw (22.57 U/mL) with G. virens, sorghum straw (16.48 U/mL) with C. sitophila, and passion fruit peel with either A. fumigatus (17.26 U/mL) or T. longibrachiatum (17.53 U/mL). The most effective intracellular i¢-fructofuranosidase activity was obtained by A. aculeatus using trub (409.46 U/mL) or passion fruit peel (44.59 U/mL). Thus, alternative carbon sources, such as orange peel and trub, exhibit great potential as inducers for the production of these enzymes. Such fungal isolates from the Atlantic Forest of Parana, Brazil are promising candidates for generating significant amounts of β-galactosidase and i¢-fructofuranosidase using abundant and inexpensive agro-industrial substrates. Key words: Agro-industrial residue, Atlantic Forest, lactase, invertase, fungus.

Cellulase and Biomass Production from Sorghum ( Sorghum guineense ) Waste by Trichoderma longibrachiatum and Aspergillus terreus

Abstract: The production of single cell protein and cellulase from cellulosic sorghum waste using Trichoderma longibrachiatum and Aspergillus terreus was assessed. Pre-treated waste was added to basal media, inoculated with test organisms and incubated at 30°C in a New Brunswick G24 gyratory shaker at 180rpm. Samples were withdrawn at intervals of 24hours for 120hours and were subjected to protein assay and mycelia weight determination. The activity of cellulose produced by each organism was determined using filter paper and carboxymethyl cellulose as substrate. Optimum pH and temperature were also determined to validate the reaction conditions of the enzymes. Trichoderma longibrachiatum and Aspergillus terreus grew well on the sorghum waste producing a mycelia weight of 0.74g and 0.81g with protein concentration of 1.63mg/ml and 0.71mg/ml at 120hours respectively. Cellulase produced by test organisms degraded filter paper and carboxymethyl cellulose with Trichoderma longibrachiatum producing an activity of 1.46U/mL and 1.82 U/mL respectively while Aspergillus terreus cellulase produced activity of 1.46U/mL and 0.95U/mL respectively. However, optimum pH for cellulase activity Trichoderma longibrachiatum and Aspergillus terreus were 5.0 and 4.0 respectively while optimal temperature for cellulase activity from the two organisms was 50°C.

Cultivation of a novel cellulase/xylanase producer, Trichoderma longibrachiatum mutant TW 1-59-27: Production of the enzyme preparation and the study of its properties

Abstract: As a result of γ-mutagenesis of Trichoderma longibrachiatum TW1 and the subsequent selection of improved producers, a novel mutant strain, TW1-59-27, capable of efficiently secreting cellulase and xylanase was obtained. In a fed-batch cultivation, the new TW1-59-27 mutant was significantly more active compared with the original TW1 strain. For instance, the activities of cellulase (towards carboxymethylcellulose) and xylanase in the culture broth (CB) increased by 1.8 and two times, respectively, and the protein content increased by 1.47 times. The activity of these enzymes in the dry enzyme preparation derived from the CB of the TW1-59-27 mutant was 1.3–1.8 times higher than that in the preparation derived from the original TW1 strain. It was established that the cellulase from the enzyme preparation of the mutant strain demonstrated the maximum activity at 55–65°C; it occurred in xylanase at 60°C. The pH optima of these enzymes were pH 4.5–5.0 and pH 5.0–6.0, respectively. It was shown that the content of endoglucanases in the enzyme preparation increased from 7% to 13.5%; the effect is largely driven by the elevated secretion of endoglucanase-1. An enzyme preparation with increased endoglucanase-1 content is promising for use as a feed additive in agriculture.

Straw composting bacterium agent as well as preparation method and application thereof

Abstract: The invention discloses a straw composting bacterium agent as well as a preparation method and application thereof and relates to the straw composting bacterium agent. The straw composting bacterium agent is compounded of 5-15 percent of bacillus subtilis powder, 50-70 percent of trichoderma longibrachiatum powder and 25-45 percent of streptomyces griseorubens powder. The preparation method comprises the following steps: respectively preparing zymophytes of bacillus subtilis, trichoderma longibrachiatum and streptomyces griseorubens and products thereof by adopting a solid fermentation method; drying and crushing to form the bacillus subtilis powder, the trichoderma longibrachiatum powder and the streptomyces griseorubens powder; mixing to obtain the straw composting bacterium agent. The straw composting bacterium agent can be applied to preparation of a straw composting and degrading agent.

Isolation and Characterization of Hydrocarbon Biodegrading Fungi from oil contaminated soils in Thika, Kenya

Abstract: Spillage and extensive exploration of petroleum products results in pollution of the environment. Bioremediation of the oil contaminated sites could be achieved by fungal biodegradation; however, the specific fungi involved have not been determined in Kenya. In this study fungal isolates from Thika in central Kenya were screened for biodegradation of engine oil. There sample size was four garages that were more than 10 years. Soil was homogenously mixed for each of the twenty seven soil samples. The initial isolation from the oil contaminated soil was done using potato dextrose agar at a temperature of 30°C for seven days. The biodegradation incubation was done using Bacto Bushnell – Haas broth at 30°C for twenty one days. Colonies were observed using a light microscope at a magnification of x1000 and characterized morphologically. The 18S rRNA genome was amplified, sequenced and the sequences used for phylogenetic analysis. The size of the amplicon targeted was 700bp in approximation. The isolates were grown at varied temperatures and pH, and screened for enzymatic activities. Analysis for the biodegraded oil was done by gas chromatography- mass spectrometry (GCHRGC 400B - MSQ12 Konic-Spain). Eight fungal isolates were recovered from polluted soils namely, Trichoderma viride, Trichoderma spirale, Neosartorya pseudofischeri, Neosartorya aureola, Aspergillus flavus, Aspergillus terreus, Penicillium griseofulvum and Trichoderma longibrachiatum. Comparison of the 18S rRNA gene sequences to known fungal sequences in the Genbank database using BLAST analysis indicated similarity of more than 97%. The percentage similarity for isolates that biodegraded oil namely, Penicillium griseofulvum and Aspergillus flavus was 99% and 100%. The eight fungi were characterized morphologically and were different from each other through different parameters like colour, elevation and margin among others. Out of eight fungal isolates recovered from contaminating soil, only Aspergillus flavus and Penicillium griseofulvum biodegraded seven and seventeen oil compounds respectively. However, some compounds could not be fully biodegraded byAspergillus flavus namely, decane, undecane and tridecane from a concentration of 5.48 to 0.23, 18.14 to 0.13 and 14.22 to 0.11 mg/l respectively. Penicillium griseofulvum could not fully biodegrade 1- Ethylidene-1–Indene from a concentration of 0.29 to 0.17mg/l. The optimum growth temperature range for the eight fungi was 30oC and 40oC. There was no growth at 50oC for all isolates except some slight growth by Aspergillus flavus. Optimum growth at pH 7 and pH 9 and poor growth at pH 5 was noted. The eight fungi produced amylase, protease, lipase/esterase and cellulase enzymes. This study will contribute to the database on locally available fungal diversity and their ecology. This will also increase knowledge of the fungi involved in biodegradation of oil in Kenya. Moreover Kenya’s Turkana has recently discovered oil and so this could be of great help in dealing with events of oil spill so as to conserve our precious environment.

Cultural Characterization of Fungi Isolated From Oil Contaminated Soils

Abstract: Application of Fungi for effective removal of hydrocarbon contamination from soil is being considered as the better option when it comes to biodegradation. Other method like physical and chemical bioremediation leads to production of toxic compounds and these methods are not cost effective. In the present study, soil samples from four different oil contaminated soils were assessed for any recovery of fungi present. Cultural characterization was used as preliminary identification using keys. Initial isolation from the oil contaminated soil was done using potato dextrose agar. Colonies were observed and characterized morphologically. The isolates were grown at varied temperatures and pH. Eight fungal isolates were recovered from polluted soils namely, Trichoderma viride, Trichoderma spirale, Neosartorya pseudofischeri, Neosartorya aureola, Aspergillus flavus, Aspergillus terreus, Penicillium griseofulvum and Trichoderma longibrachiatum . The optimum growth temperature range for the eight fungi was 30 o C and 40 o C. There was no growth at 50 o C for all isolates except some slight growth by Aspergillus flavus . Optimum growth at pH 7 and pH 9 and poor growth at pH 5 was noted. This study will contribute to the database on locally available fungal diversity and their ecology. Keywords . Fungi, Bioremediation, oil contamination, biodegradation

Trichoderma for preventing and controlling cotton verticillium wilt and application of trichoderma

Abstract: The invention provides Trichoderma longibrachiatum with a preservation number of CGMCC No.9727. The invention further provides a compound microorganism microbial agent. The compound microorganism microbial agent comprises the following active ingredients: Trichoderma longibrachiatum, Bacillus cereus and Bacillus mojavensis. The invention further provides application of Trichoderma longibrachiatum or the compound microorganism microbial agent in prevention and treatment of cotton verticillium wilt. According to the technical scheme, the cotton verticillium wilt can be effectively prevented and controlled.

Trichoderma longibrachiatum thica-4 strain acting as a fungicide and promoting the growth of plants of agricultural interest.

Abstract: The present invention refers to a purified strain of Trichoderma longibrachiatum identified as ThlCA-4 based on molecular analysis with code GenBank: HQ667667.1, characterized in that it has the sequence SEQ:ID:1 and is deposited in the Agricultural Research Culture Collection (NRRL) with the code: NRRL50762. The microorganism is a mycoparasite fungus with fungicide properties of wide spectrum, which produces substances for promoting the vegetable growth upon being inoculated in horticultural interest plants, mainly transgenic or conventional cotton and cucumber. This organism is adapted to the environmental conditions of the Valley of Mexicali, Baja Califormia, were it was isolated and identified.