Trichoderma harzianum enhances the production of nematicidal compounds in vitro and improves biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato.

PubMed

Siddiqui, I A; Shaukat, S S

2004-01-01

To determine the influence of soil-borne fungus Trichoderma harzianum on the biocontrol performance of Pseudomonas fluorescens strain CHA0 and its 2,4-diacetylphloroglucinol (DAPG) overproducing derivative CHA0/pME3424 against Meloidogyne javanica. Amendment of the culture filtrate (CF) or methanol extract of the CF of a T. harzianum strain Th6 to P. fluorescens growth medium enhanced the production of nematicidal compound(s) by bacterial inoculants in vitro. In addition, bacteria overwhelmingly expressed phl'-'lacZ reporter gene when the medium was amended with CF of T. harzianum. Pseudomonas fluorescens and T. harzianum applied together in unsterilized sandy loam soil caused greater reduction in nematode population densities in tomato roots. Trichoderma harzianum improves root-knot nematode biocontrol by the antagonistic rhizobacterium P. fluorescens both in vitro and under glasshouse conditions. The synergistic effect of T. harzianum on the production of nematicidal compound(s) critical in biocontrol may improve the efficacy of biocontrol bacteria against plant-parasitic nematodes. Considering the inconsistent performance of the biocontrol agents under field conditions, application of a mixture of compatible T. harzianum and P. fluorescens would more closely mimic the natural situation and might broaden the spectrum of biocontrol activity with enhanced efficacy and reliability of control.

The relation between xyr1 overexpression in Trichoderma harzianum and sugarcane bagasse saccharification performance.

PubMed

da Silva Delabona, Priscila; Rodrigues, Gisele Nunes; Zubieta, Mariane Paludetti; Ramoni, Jonas; Codima, Carla Aloia; Lima, Deise Juliana; Farinas, Cristiane Sanchez; da Cruz Pradella, José Geraldo; Seiboth, Bernhard

2017-03-20

This work investigates the influence of the positive regulator XYR1 of Trichoderma harzianum on the production of cellulolytic enzymes, using sugarcane bagasse as carbon source. Constitutive expression of xyr1 was achieved under the control of the strong Trichoderma reesei pki1 promoter. Five clones with xyr1 overexpression achieved higher xyr1 expression and greater enzymatic productivity when cultivated under submerged fermentation, hence validating the genetic construction for T. harzianum. Clone 5 presented a relative expression of xyr1 26-fold higher than the parent strain and exhibited 66, 37, and 36% higher values for filter paper activity, xylanase activity, and β-glucosidase activity, respectively, during cultivation in a stirred-tank bioreactor. The overexpression of xyr1 in T. harzianum resulted in an enzymatic complex with significantly improved performance in sugarcane bagasse saccharification, with an enhancement of 25% in the first 24h. Our results also show that constitutive overexpression of xyr1 leads to the induction of several important players in biomass degradation at early (24h) and also late (48h) timepoints of inoculation. However, we also observed that the carbon catabolite repressor CRE1 was upregulated in xyr1 overexpression mutants. These findings demonstrate the feasibility of improving cellulase production by modifying regulator expression and suggest an attractive approach for increasing total cellulase productivity in T. harzianum. Copyright © 2017 Elsevier B.V. All rights reserved.

40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2 (ATCC...

40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2 (ATCC...

40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2 (ATCC...

40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2 (ATCC...

40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2 (ATCC...

Secretome analysis of the fungus Trichoderma harzianum grown on cellulose.

PubMed

Do Vale, Luis H F; Gómez-Mendoza, Diana P; Kim, Min-Sik; Pandey, Akhilesh; Ricart, Carlos A O; Ximenes F Filho, Edivaldo; Sousa, Marcelo V

2012-08-01

Trichoderma harzianum is a mycoparasitic filamentous fungus that produces and secretes a wide range of extracellular hydrolytic enzymes used in cell wall degradation. Due to its potential in biomass conversion, T. harzianum draws great attention from biofuel and biocontrol industries and research. Here, we report an extensive secretome analysis of T. harzianum. The fungus was grown on cellulose medium, and its secretome was analyzed by a combination of enzymology, 2DE, MALDI-MS and -MS/MS (Autoflex II), and LC-MS/MS (LTQ-Orbitrap XL). A total of 56 proteins were identified using high-resolution MS. Interestingly, although cellulases were found, the major hydrolytic enzymes secreted in the cellulose medium were chitinases and endochitinases, which may reflect the biocontrol feature of T. harzianum. The glycoside hydrolase family, including chitinases (EC 3.2.1.14), endo-N-acetylglucosaminidases (EC 3.2.1.96), hexosaminidases (EC 3.2.1.52), galactosidases (EC 3.2.1.23), xylanases (EC 3.2.1.8), exo-1,3-glucanases (EC 3.2.1.58), endoglucanases (EC 3.2.1.4), xylosidases (EC 3.2.1.37), α-L-arabinofuranosidase (EC 3.2.1.55), N-acetylhexosaminidases (EC 3.2.1.52), and other enzymes represented 51.36% of the total secretome. Few representatives were classified in the protease family (8.90%). Others (17.60%) are mostly intracellular proteins. A considerable part of the secretome was composed of hypothetical proteins (22.14%), probably because of the absence of an annotated T. harzianum genome. The T. harzianum secretome composition highlights the importance of this fungus as a rich source of hydrolytic enzymes for bioconversion and biocontrol applications. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Desorption of zinc by extracellularly produced metabolites of Trichoderma harzianum, Trichoderma reesei and Coriolus versicolor.

PubMed

Adams, P; Lynch, J M; De Leij, F A A M

2007-12-01

To determine the role of fungal metabolites in the desorption of metals. Desorption of Zn from charcoal by three different fungi was compared against metal desorption with reverse osmosis water, a 0.1% Tween 80 solution and a 0.1 mol l(-1) CaCl(2) solution. All three fungal filtrates desorbed three times more Zn than either 0.1% Tween 80 or 0.1 mol l(-1) CaCl(2). Metal chelator production in Trichoderma harzianum and Coriolus versicolor was constitutively expressed while chelator production in Trichoderma reesei was induced by Zn. The presence of Zn inhibited the production of metal chelators by C. versicolor. Only C. versicolor was found to produce oxalic acid (a strong metal chelator). All fungi caused a marked decrease in pH, although this was not enough to explain the increased desorption of the metals by the different fungal filtrates. Metal chelation via organic acids and proteins are the main mechanisms by which the fungal filtrates increase zinc desorption. The results of this study explain why plants inoculated with T. harzianum T22 take up more metal from soil, than noninoculated plants while metabolites produced by fungi could be used for metal leaching from contaminated soils.

Synergistic effects of plant defense elicitors and Trichoderma harzianum on enhanced induction of antioxidant defense system in tomato against Fusarium wilt disease.

PubMed

Zehra, Andleeb; Meena, Mukesh; Dubey, Manish Kumar; Aamir, Mohd; Upadhyay, R S

2017-11-02

Plant defense against their pathogens can be induced by a complex network of different inducers. The present study investigates the synergistic effect of Trichoderma harzianum, exogenous salicylic acid (SA) and methyl jasmonate (MeJA) over the response and regulation of the antioxidant defense mechanisms and lipid peroxidation in tomato plants against Fusarium wilt disease. In the present work, tomato plants were infected by Fusarium oxysporum f. sp. lycopersici 3 days after inoculated with T. harzianum and/or sprayed daily for 3 days with chemical inducers (SA and MeJA). Plants were analysed at 0, 24, 48, 72 and 96 h after inoculation with Fusarium oxysporum f. sp. lycopersici. Infection of tomato plants by pathogen led to strong reduction in the dry weight of roots and shoots with the enhanced concentration of H 2 O 2 and varying degree of lipid peroxidation. Concurrently, exogenous SA, when applied with pathogen greatly enhanced H 2 O 2 content as well as activities of antioxidant enzymes except catalase (CAT) and ascorbate peroxidase (APx). The pathogen challenged plants pretreated with T. harzianum and MeJA together exhibited less lipid peroxidation and as well as the elevated level of ascorbic acid and enhanced activities of antioxidant enzymes. All applied treatments protected tomato seedlings against Fusarium wilt disease but the percentage of protection was found higher in plants pretreated with the combination of T. harzianum and chemical inducers.

Differential expression of oil palm pathology genes during interactions with Ganoderma boninense and Trichoderma harzianum.

PubMed

Alizadeh, Fahimeh; Abdullah, Siti Nor Akmar; Khodavandi, Alireza; Abdullah, Faridah; Yusuf, Umi Kalsom; Chong, Pei Pei

2011-07-01

The expression profiles of Δ9 stearoyl-acyl carrier protein desaturase (SAD1 and SAD2) and type 3 metallothionein (MT3-A and MT3-B) were investigated in seedlings of oil palm (Elaeis guineensis) artificially inoculated with the pathogenic fungus Ganoderma boninense and the symbiotic fungus Trichoderma harzianum. Expression of SAD1 and MT3-A in roots and SAD2 in leaves were significantly up-regulated in G. boninense inoculated seedlings at 21 d after treatment when physical symptoms had not yet appeared and thereafter decreased to basal levels when symptoms became visible. Our finding demonstrated that the SAD1 expression in leaves was significantly down-regulated to negligible levels at 42 and 63 d after treatment. The transcripts of MT3 genes were synthesized in G. boninense inoculated leaves at 42 d after treatment, and the analyses did not show detectable expression of these genes before 42 d after treatment. In T. harzianum inoculated seedlings, the expression levels of SAD1 and SAD2 increased gradually and were stronger in roots than leaves, while for MT3-A and MT3-B, the expression levels were induced in leaves at 3d after treatment and subsequently maintained at same levels until 63d after treatment. The MT3-A expression was significantly up-regulated in roots at 3d after treatment and thereafter were maintained at this level. Both SAD and MT3 expression were maintained at maximum levels or at levels higher than basal. This study demonstrates that oil palm was able to distinguish between pathogenic and symbiotic fungal interactions, thus resulting in different transcriptional activation profiles of SAD and MT3 genes. Increases in expression levels of SAD and MT3 would lead to enhanced resistance against G. boninense and down-regulation of genes confer potential for invasive growth of the pathogen. Differences in expression profiles of SAD and MT3 relate to plant resistance mechanisms while supporting growth enhancing effects of symbiotic T. harzianum

Greenhouse evaluation of Bacillus subtilis AP-01 and Trichoderma harzianum AP-001 in controlling tobacco diseases.

PubMed

Maketon, Monchan; Apisitsantikul, Jirasak; Siriraweekul, Chatchai

2008-04-01

Two biological control agents, Bacillus subtilis AP-01 (Larminar(™)) and Trichoderma harzianum AP-001 (Trisan(™)) alone or/in combination were investigated in controlling three tobacco diseases, including bacterial wilt (Ralstonia solanacearum), damping-off (Pythium aphanidermatum), and frogeye leaf spot (Cercospora nicotiana). Tests were performed in greenhouse by soil sterilization prior to inoculation of the pathogens. Bacterial-wilt and damping off pathogens were drenched first and followed with the biological control agents and for comparison purposes, two chemical fungicides. But for frogeye leaf spot, which is an airborne fungus, a spraying procedure for every treatment including a chemical fungicide was applied instead of drenching. Results showed that neither B. subtilis AP-01 nor T harzianum AP-001 alone could control the bacterial wilt, but when combined, their controlling capabilities were as effective as a chemical treatment. These results were also similar for damping-off disease when used in combination. In addition, the combined B. subtilis AP-01 and T. harzianum AP-001 resulted in a good frogeye leaf spot control, which was not significantly different from the chemical treatment.

The platelet-activating factor acetylhydrolase gene derived from Trichoderma harzianum induces maize resistance to Curvularia lunata through the jasmonic acid signaling pathway.

PubMed

Yu, Chuanjin; Fan, Lili; Gao, Jinxin; Wang, Meng; Wu, Qiong; Tang, Jun; Li, Yaqian; Chen, Jie

2015-01-01

Platelet-activating factor acetylhydrolase (PAF-AH) derived from Trichoderma harzianum was upregulated by the interaction of T. harzianum with maize roots or the foliar pathogen Curvularia lunata. PAF-AH was associated with chitinase and cellulase expressions, but especially with chitinase, because its activity in the KO40 transformant (PAF-AH disruption transformant) was lower, compared with the wild-type strain T28. The result demonstrated that the colonization of maize roots by T. harzianum induced systemic protection of leaves inoculated with C. lunata. Such protection was associated with the expression of inducible jasmonic acid pathway-related genes. Moreover, the data from liquid chromatography-mass spectrometry confirmed that the concentration of jasmonic acid in maize leaves was associated with the expression level of defense-related genes, suggesting that PAF-AH induced resistance to the foliar pathogen. Our findings showed that PAF-AH had an important function in inducing systemic resistance to maize leaf spot pathogen.

Downy mildew resistance induced by Trichoderma harzianum T39 in susceptible grapevines partially mimics transcriptional changes of resistant genotypes

PubMed Central

2012-01-01

Background Downy mildew, caused by Plasmopara viticola, is one of the most severe diseases of grapevine and is commonly controlled by fungicide treatments. The beneficial microorganism Trichoderma harzianum T39 (T39) can induce resistance to downy mildew, although the molecular events associated with this process have not yet been elucidated in grapevine. A next generation RNA sequencing (RNA-Seq) approach was used to study global transcriptional changes associated with resistance induced by T39 in Vitis vinifera Pinot Noir leaves. The long-term aim was to develop strategies to optimize the use of this agent for downy mildew control. Results More than 14.8 million paired-end reads were obtained for each biological replicate of T39-treated and control leaf samples collected before and 24 h after P. viticola inoculation. RNA-Seq analysis resulted in the identification of 7,024 differentially expressed genes, highlighting the complex transcriptional reprogramming of grapevine leaves during resistance induction and in response to pathogen inoculation. Our data show that T39 has a dual effect: it directly modulates genes related to the microbial recognition machinery, and it enhances the expression of defence-related processes after pathogen inoculation. Whereas several genes were commonly affected by P. viticola in control and T39-treated plants, opposing modulation of genes related to responses to stress and protein metabolism was found. T39-induced resistance partially inhibited some disease-related processes and specifically activated defence responses after P. viticola inoculation, causing a significant reduction of downy mildew symptoms. Conclusions The global transcriptional analysis revealed that defence processes known to be implicated in the reaction of resistant genotypes to downy mildew were partially activated by T39-induced resistance in susceptible grapevines. Genes identified in this work are an important source of markers for selecting novel

Trichoderma harzianum enhances tomato indirect defense against aphids.

PubMed

Coppola, Mariangela; Cascone, Pasquale; Chiusano, Maria Luisa; Colantuono, Chiara; Lorito, Matteo; Pennacchio, Francesco; Rao, Rosa; Woo, Sheridan Lois; Guerrieri, Emilio; Digilio, Maria Cristina

2017-12-01

Many fungal root symbionts of the genus Trichoderma are well-known for their beneficial effects on agronomic performance and protection against plant pathogens; moreover, they may enhance protection from insect pests, by triggering plant resistance mechanisms. Defense barriers against insects are induced by the activation of metabolic pathways involved in the production of defense-related plant compounds, either directly active against herbivore insects, or exerting an indirect effect, by increasing the attraction of herbivore natural enemies. In a model system composed of the tomato plant, the aphid Macrosiphum euphorbiae and the parasitoid Aphidius ervi, plant metabolic changes induced by Trichoderma harzianum and their effects on higher trophic levels have been assessed. T. harzianum T22 treatments induce a primed state that upon aphid attacks leads to an increased attraction of aphid parasitoids, mediated by the enhanced production of volatile organic compounds (VOCs) that are known to induce Aphidius ervi flight. Transcriptome sequencing of T22-treated plants infested by aphids showed a remarkable upregulation of genes involved in terpenoids biosynthesis and salicylic acid pathway, which are consistent with the observed flight response of A. ervi and the VOC bouquet profile underlying this behavioral response. © 2017 Institute of Zoology, Chinese Academy of Sciences.

Involvement of Trichoderma harzianum Epl-1 Protein in the Regulation of Botrytis Virulence- and Tomato Defense-Related Genes.

PubMed

Gomes, Eriston V; Ulhoa, Cirano J; Cardoza, Rosa E; Silva, Roberto N; Gutiérrez, Santiago

2017-01-01

Several Trichoderma spp. are well known for their ability to: (i) act as important biocontrol agents against phytopathogenic fungi; (ii) function as biofertilizers; (iii) increase the tolerance of plants to biotic and abiotic stresses; and (iv) induce plant defense responses via the production and secretion of elicitor molecules. In this study, we analyzed the gene-regulation effects of Trichoderma harzianum Epl-1 protein during the interactions of mutant Δ epl-1 or wild-type T. harzianum strains with: (a) the phytopathogen Botrytis cinerea and (b) with tomato plants, on short (24 h hydroponic cultures) and long periods (4-weeks old plants) after Trichoderma inoculation. Our results indicate that T. harzianum Epl-1 protein affects the in vitro expression of B. cinerea virulence genes, especially those involved in the botrydial biosynthesis ( BcBOT genes), during the mycoparasitism interaction. The tomato defense-related genes were also affected, indicating that Epl-1 is involved in the elicitation of the salicylic acid pathway. Moreover, Epl-1 also regulates the priming effect in host tomato plants and contributes to enhance the interaction with the host tomato plant during the early stage of root colonization.

Involvement of Trichoderma harzianum Epl-1 Protein in the Regulation of Botrytis Virulence- and Tomato Defense-Related Genes

PubMed Central

Gomes, Eriston V.; Ulhoa, Cirano J.; Cardoza, Rosa E.; Silva, Roberto N.; Gutiérrez, Santiago

2017-01-01

Several Trichoderma spp. are well known for their ability to: (i) act as important biocontrol agents against phytopathogenic fungi; (ii) function as biofertilizers; (iii) increase the tolerance of plants to biotic and abiotic stresses; and (iv) induce plant defense responses via the production and secretion of elicitor molecules. In this study, we analyzed the gene-regulation effects of Trichoderma harzianum Epl-1 protein during the interactions of mutant Δepl-1 or wild-type T. harzianum strains with: (a) the phytopathogen Botrytis cinerea and (b) with tomato plants, on short (24 h hydroponic cultures) and long periods (4-weeks old plants) after Trichoderma inoculation. Our results indicate that T. harzianum Epl-1 protein affects the in vitro expression of B. cinerea virulence genes, especially those involved in the botrydial biosynthesis (BcBOT genes), during the mycoparasitism interaction. The tomato defense-related genes were also affected, indicating that Epl-1 is involved in the elicitation of the salicylic acid pathway. Moreover, Epl-1 also regulates the priming effect in host tomato plants and contributes to enhance the interaction with the host tomato plant during the early stage of root colonization. PMID:28611802

Cloning, annotation and expression analysis of mycoparasitism-related genes in Trichoderma harzianum 88.

PubMed

Yao, Lin; Yang, Qian; Song, Jinzhu; Tan, Chong; Guo, Changhong; Wang, Li; Qu, Lianhai; Wang, Yun

2013-04-01

Trichoderma harzianum 88, a filamentous soil fungus, is an effective biocontrol agent against several plant pathogens. High-throughput sequencing was used here to study the mycoparasitism mechanisms of T. harzianum 88. Plate confrontation tests of T. harzianum 88 against plant pathogens were conducted, and a cDNA library was constructed from T. harzianum 88 mycelia in the presence of plant pathogen cell walls. Randomly selected transcripts from the cDNA library were compared with eukaryotic plant and fungal genomes. Of the 1,386 transcripts sequenced, the most abundant Gene Ontology (GO) classification group was "physiological process". Differential expression of 19 genes was confirmed by real-time RT-PCR at different mycoparasitism stages against plant pathogens. Gene expression analysis revealed the transcription of various genes involved in mycoparasitism of T. harzianum 88. Our study provides helpful insights into the mechanisms of T. harzianum 88-plant pathogen interactions.

Involvement of pachybasin and emodin in self-regulation of Trichoderma harzianum mycoparasitic coiling.

PubMed

Lin, Yi-Ruu; Lo, Chaur-Tsuen; Liu, Shu-Ying; Peng, Kou-Cheng

2012-03-07

Our aim was to determine the effects of two secondary metabolites secreted by Trichoderma harzianum, pachybasin and emodin, on the mycoparasitic coiling behavior and cAMP content of T. harzianum. The number of T. harzianum coils around Nylon 66 fiber was increased in the presence of R. solani. The number of T. harzianum coils around R. solani hyphae and Nylon 66 fiber were significantly increased in the presence of pachybasin and emodin. The cAMP level in T. harzianum was significantly increased by close contact with R. solani and much higer cAMP level in the presence of exogenous pachybasin and emodin. A cAMP inhibitor diminished the effect of pachybasin and emodin on T. harzianum coiling around Nylon 66 fiber. The results suggest that pachybasin and emodin mediate the increase in the number of Trichoderma mycoparasitic coils via cAMP signaling. This is the first report to suggest that pachybasin and emodin play roles in the biocontrol mechanism of Trichoderma.

Trichoderma harzianum MTCC 5179 impacts the population and functional dynamics of microbial community in the rhizosphere of black pepper (Piper nigrum L.).

PubMed

Umadevi, Palaniyandi; Anandaraj, Muthuswamy; Srivastav, Vivek; Benjamin, Sailas

2017-11-29

Employing Illumina Hiseq whole genome metagenome sequencing approach, we studied the impact of Trichoderma harzianum on altering the microbial community and its functional dynamics in the rhizhosphere soil of black pepper (Piper nigrum L.). The metagenomic datasets from the rhizosphere with (treatment) and without (control) T. harzianum inoculation were annotated using dual approach, i.e., stand alone and MG-RAST. The probiotic application of T. harzianum in the rhizhosphere soil of black pepper impacted the population dynamics of rhizosphere bacteria, archae, eukaryote as reflected through the selective recruitment of bacteria [Acidobacteriaceae bacterium (p=1.24e-12), Candidatus koribacter versatilis (p=2.66e-10)] and fungi [(Fusarium oxysporum (p=0.013), Talaromyces stipitatus (p=0.219) and Pestalotiopsis fici (p=0.443)] in terms of abundance in population and bacterial chemotaxis (p=0.012), iron metabolism (p=2.97e-5) with the reduction in abundance for pathogenicity islands (p=7.30e-3), phages and prophages (p=7.30e-3) with regard to functional abundance. Interestingly, it was found that the enriched functional metagenomic signatures on phytoremediation such as benzoate transport and degradation (p=2.34e-4), and degradation of heterocyclic aromatic compounds (p=3.59e-13) in the treatment influenced the rhizosphere micro ecosystem favoring growth and health of pepper plant. The population dynamics and functional richness of rhizosphere ecosystem in black pepper influenced by the treatment with T. harzianum provides the ecological importance of T. harzianum in the cultivation of black pepper. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Dose-dependent response of Trichoderma harzianum in improving drought tolerance in rice genotypes.

PubMed

Pandey, Veena; Ansari, Mohammad W; Tula, Suresh; Yadav, Sandep; Sahoo, Ranjan K; Shukla, Nandini; Bains, Gurdeep; Badal, Shail; Chandra, Subhash; Gaur, A K; Kumar, Atul; Shukla, Alok; Kumar, J; Tuteja, Narendra

2016-05-01

This study demonstrates a dose-dependent response of Trichoderma harzianum Th-56 in improving drought tolerance in rice by modulating proline, SOD, lipid peroxidation product and DHN / AQU transcript level, and the growth attributes. In the present study, the effect of colonization of different doses of T. harzianum Th-56 strain in rice genotypes were evaluated under drought stress. The rice genotypes treated with increasing dose of T. harzianum strain Th-56 showed better drought tolerance as compared with untreated control plant. There was significant change in malondialdehyde, proline, higher superoxide dismutase level, plant height, total dry matter, relative chlorophyll content, leaf rolling, leaf tip burn, and the number of scorched/senesced leaves in T. harzianum Th-56 treated rice genotypes under drought stress. This was corroborated with altered expression of aquaporin and dehydrin genes in T. harzianum Th-56 treated rice genotypes. The present findings suggest that a dose of 30 g/L was the most effective in improving drought tolerance in rice, and its potential exploitation will contribute to the advancement of rice genotypes to sustain crop productivity under drought stress. Interaction studies of T. harzianum with three aromatic rice genotypes suggested that PSD-17 was highly benefitted from T. harzianum colonization under drought stress.

Carbohydrate-active enzymes in Trichoderma harzianum: a bioinformatic analysis bioprospecting for key enzymes for the biofuels industry.

PubMed

Ferreira Filho, Jaire Alves; Horta, Maria Augusta Crivelente; Beloti, Lilian Luzia; Dos Santos, Clelton Aparecido; de Souza, Anete Pereira

2017-10-12

Trichoderma harzianum is used in biotechnology applications due to its ability to produce powerful enzymes for the conversion of lignocellulosic substrates into soluble sugars. Active enzymes involved in carbohydrate metabolism are defined as carbohydrate-active enzymes (CAZymes), and the most abundant family in the CAZy database is the glycoside hydrolases. The enzymes of this family play a fundamental role in the decomposition of plant biomass. In this study, the CAZymes of T. harzianum were identified and classified using bioinformatic approaches after which the expression profiles of all annotated CAZymes were assessed via RNA-Seq, and a phylogenetic analysis was performed. A total of 430 CAZymes (3.7% of the total proteins for this organism) were annotated in T. harzianum, including 259 glycoside hydrolases (GHs), 101 glycosyl transferases (GTs), 6 polysaccharide lyases (PLs), 22 carbohydrate esterases (CEs), 42 auxiliary activities (AAs) and 46 carbohydrate-binding modules (CBMs). Among the identified T. harzianum CAZymes, 47% were predicted to harbor a signal peptide sequence and were therefore classified as secreted proteins. The GH families were the CAZyme class with the greatest number of expressed genes, including GH18 (23 genes), GH3 (17 genes), GH16 (16 genes), GH2 (13 genes) and GH5 (12 genes). A phylogenetic analysis of the proteins in the AA9/GH61, CE5 and GH55 families showed high functional variation among the proteins. Identifying the main proteins used by T. harzianum for biomass degradation can ensure new advances in the biofuel production field. Herein, we annotated and characterized the expression levels of all of the CAZymes from T. harzianum, which may contribute to future studies focusing on the functional and structural characterization of the identified proteins.

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

PubMed Central

Pang, Guan; Shen, Qi-Rong; Li, Rong; Chen, Wei

2015-01-01

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 (i.e., 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

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

PubMed

Li, Rui-Xia; Cai, Feng; Pang, Guan; Shen, Qi-Rong; Li, Rong; Chen, Wei

2015-01-01

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 (i.e., 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

qRT-PCR quantification of the biological control agent Trichoderma harzianum in peat and compost-based growing media.

PubMed

Beaulieu, Robert; López-Mondéjar, Rubén; Tittarelli, Fabio; Ros, Margarita; Pascual, José Antonio

2011-02-01

To ensure proper use of Trichoderma harzianum in agriculture, accurate data must be obtained in population monitoring. The effectiveness of qRT-PCR to quantify T. harzianum in different growing media was compared to the commonly used techniques of colony counting and qPCR. Results showed that plate counting and qPCR offered similar T. harzianum quantification patterns of an initial rapid increase in fungal population that decreased over time. However, data from qRT-PCR showed a population curve of active T. harzianum with a delayed onset of initial growth which then increased throughout the experiment. Results demonstrated that T. harzianum can successfully grow in these media and that qRT-PCR can offer a more distinct representation of active T. harzianum populations. Additionally, compost amended with T. harzianum exhibited a lower Fusarium oxysporum infection rate (67%) and lower percentage of fresh weight loss (11%) in comparison to amended peat (90% infection rate, 23% fresh weight loss). Copyright © 2010 Elsevier Ltd. All rights reserved.

Trichoderma harzianum might impact phosphorus transport by arbuscular mycorrhizal fungi.

PubMed

De Jaeger, Nathalie; de la Providencia, Ivan E; de Boulois, Hervé Dupré; Declerck, Stéphane

2011-09-01

Trichoderma sp. is a biocontrol agent active against plant pathogens via mechanisms such as mycoparasitism. Recently, it was demonstrated that Trichoderma harzianum was able to parasitize the mycelium of an arbuscular mycorrhizal (AM) fungus, thus affecting its viability. Here, we question whether this mycoparasitism may reduce the capacity of Glomus sp. to transport phosphorus ((33)P) to its host plant in an in vitro culture system. (33)P was measured in the plant and in the fungal mycelium in the presence/absence of T. harzianum. The viability and metabolic activity of the extraradical mycelium was measured via succinate dehydrogenase and alkaline phosphatase staining. Our study demonstrated an increased uptake of (33)P by the AM fungus in the presence of T. harzianum, possibly related to a stress reaction caused by mycoparasitism. In addition, the disruption of AM extraradical hyphae in the presence of T. harzianum affected the (33)P translocation within the AM fungal mycelium and consequently the transfer of (33)P to the host plant. The effects of T. harzianum on Glomus sp. may thus impact the growth and function of AM fungi and also indirectly plant performance by influencing the source-sink relationship between the two partners of the symbiosis. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Isolation and expression of two polyketide synthase genes from Trichoderma harzianum 88 during mycoparasitism.

PubMed

Yao, Lin; Tan, Chong; Song, Jinzhu; Yang, Qian; Yu, Lijie; Li, Xinling

2016-01-01

Metabolites of mycoparasitic fungal species such as Trichoderma harzianum 88 have important biological roles. In this study, two new ketoacyl synthase (KS) fragments were isolated from cultured Trichoderma harzianum 88 mycelia using degenerate primers and analysed using a phylogenetic tree. The gene fragments were determined to be present as single copies in Trichoderma harzianum 88 through southern blot analysis using digoxigenin-labelled KS gene fragments as probes. The complete sequence analysis in formation of pksT-1 (5669bp) and pksT-2 (7901bp) suggests that pksT-1 exhibited features of a non-reducing type I fungal PKS, whereas pksT-2 exhibited features of a highly reducing type I fungal PKS. Reverse transcription polymerase chain reaction indicated that the isolated genes are differentially regulated in Trichoderma harzianum 88 during challenge with three fungal plant pathogens, which suggests that they participate in the response of Trichoderma harzianum 88 to fungal plant pathogens. Furthermore, disruption of the pksT-2 encoding ketosynthase-acyltransferase domains through Agrobacterium-mediated gene transformation indicated that pksT-2 is a key factor for conidial pigmentation in Trichoderma harzianum 88. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Proteomic analysis of grapevine resistance induced by Trichoderma harzianum T39 reveals specific defence pathways activated against downy mildew

PubMed Central

Perazzolli, Michele

2012-01-01

Downy mildew is caused by the oomycete Plasmopara viticola and is one of the most serious diseases of grapevine. The beneficial microorganism Trichoderma harzianum T39 (T39) has previously been shown to induce plant-mediated resistance and to reduce the severity of downy mildew in susceptible grapevines. In order to better understand the cellular processes associated with T39-induced resistance, the proteomic and histochemical changes activated by T39 in grapevine were investigated before and 1 day after P. viticola inoculation. A comprehensive proteomic analysis of T39-induced resistance in grapevine was performed using an eight-plex iTRAQ protocol, resulting in the identification and quantification of a total of 800 proteins. Most of the proteins directly affected by T39 were found to be involved in signal transduction, indicating activation of a complete microbial recognition machinery. Moreover, T39-induced resistance was associated with rapid accumulation of reactive oxygen species and callose at infection sites, as well as changes in abundance of proteins involved in response to stress and redox balance, indicating an active defence response to downy mildew. On the other hand, proteins affected by P. viticola in control plants mainly decreased in abundance, possibly reflecting the establishment of a compatible interaction. Finally, the high-throughput iTRAQ protocol allowed de novo peptide sequencing, which will be used to improve annotation of the Vitis vinifera cv. Pinot Noir proteome. PMID:23105132

Biological effects of Trichoderma harzianum peptaibols on mammalian cells.

PubMed

Peltola, Joanna; Ritieni, Alberto; Mikkola, Raimo; Grigoriev, Pavel A; Pócsfalvi, Gabriella; Andersson, Maria A; Salkinoja-Salonen, Mirja S

2004-08-01

Trichoderma species isolated from water-damaged buildings were screened for toxicity by using boar sperm cells as indicator cells. The crude methanolic cell extract from Trichoderma harzianum strain ES39 inhibited the boar sperm cell motility at a low exposure concentration (50% effective concentration, 1 to 5 microg [dry weight] ml of extended boar semen(-1)). The same exposure concentration depleted the boar sperm cells of NADH(2). Inspection of the exposed boar sperm cells by transmission electron microscopy revealed damage to the plasma membrane. By using the black lipid membrane technique, it was shown that the semipurified metabolites (eluted from a SepPak C(18) cartridge) of T. harzianum strain ES39 induced voltage-dependent conductivity. The high-performance liquid chromatography-purified metabolites of T. harzianum strain ES39 dissipated the mitochondrial membrane potential (Deltapsi(m)) of human lung epithelial carcinoma cells (cell line A549). The semipurified metabolites (eluted from a SepPak C(18) cartridge) of T. harzianum strain ES39 were analyzed by mass spectrometry (MS). Matrix-assisted laser desorption ionization and nanoflow electrospray ionization MS revealed five major peptaibols, each of which contained 18 residues and had a mass ranging from 1,719 to 1,775 Da. Their partial amino acid sequences were determined by collision-induced dissociation tandem MS.

Trichoderma harzianum: Inhibition of mycotoxin producing fungi and toxin biosynthesis.

PubMed

Braun, H; Woitsch, L; Hetzer, B; Geisen, R; Zange, B; Schmidt-Heydt, M

2018-04-19

A quarter of the world-wide crop is spoiled by filamentous fungi and their mycotoxins and weather extremes associated with the climate change lead to further deterioration of the situation. The ingestion of mycotoxins causes several health issues leading in the worst case to cancer in humans and animals. Common intervention strategies against mycotoxin producing fungi, such as the application of fungicides, may result in undesirable residues and in some cases to a stress induction of mycotoxin biosynthesis. Moreover, development of fungicide resistances has greatly impacted pre- and postharvest fungal diseases. Hence there is the need to develop alternative strategies to reduce fungal infestation and thus mycotoxin contamination in the food chain. Such a strategy for natural competition of important plant-pathogenic and mycotoxin producing fungi could be Trichoderma harzianum, a mycoparasitic fungus. Especially in direct comparison to certain tested fungicides, the inhibition of different tested fungal species by T. harzianum was comparable, more sustainable and in some cases more effective, too. Besides substantially reduced growth rates, a transcriptional based inhibition of mycotoxin biosynthesis in the competed Aspergillus species could be shown. Furthermore it could be clearly observed by high-resolution Scanning Electron Microscopy (SEM) that T. harzianum actively attaches to the competitor species followed by subsequent enzymatic lysis of those mycelial filaments. The analyzed isolate of T. harzianum MRI349 is not known to produce mycotoxins. In this study it could be successfully proven that T. harzianum as a biological competitor is an effective complement to the use of fungicides. Copyright © 2018 Elsevier B.V. All rights reserved.

Degradation of polyethylene by Trichoderma harzianum--SEM, FTIR, and NMR analyses.

PubMed

Sowmya, H V; Ramalingappa; Krishnappa, M; Thippeswamy, B

2014-10-01

Trichoderma harzianum was isolated from local dumpsites of Shivamogga District for use in the biodegradation of polyethylene. Soil sample of that dumpsite was used for isolation of T. harzianum. Degradation was carried out using autoclaved, UV-treated, and surface-sterilized polyethylene. Degradation was monitored by observing weight loss and changes in physical structure by scanning electron microscopy, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. T. harzianum was able to degrade treated polyethylene (40%) more efficiently than autoclaved (23%) and surface-sterilized polyethylene (13%). Enzymes responsible for polyethylene degradation were screened from T. harzianum and were identified as laccase and manganese peroxidase. These enzymes were produced in large amount, and their activity was calculated using spectrophotometric method and crude extraction of enzymes was carried out. Molecular weight of laccase was determined as 88 kDa and that of manganese peroxidase was 55 kDa. The capacity of crude enzymes to degrade polyethylene was also determined. By observing these results, we can conclude that this organism may act as solution for the problem caused by polyethylene in nature.

Biological Effects of Trichoderma harzianum Peptaibols on Mammalian Cells

PubMed Central

Peltola, Joanna; Ritieni, Alberto; Mikkola, Raimo; Grigoriev, Pavel A.; Pócsfalvi, Gabriella; Andersson, Maria A.; Salkinoja-Salonen, Mirja S.

2004-01-01

Trichoderma species isolated from water-damaged buildings were screened for toxicity by using boar sperm cells as indicator cells. The crude methanolic cell extract from Trichoderma harzianum strain ES39 inhibited the boar sperm cell motility at a low exposure concentration (50% effective concentration, 1 to 5 μg [dry weight] ml of extended boar semen−1). The same exposure concentration depleted the boar sperm cells of NADH2. Inspection of the exposed boar sperm cells by transmission electron microscopy revealed damage to the plasma membrane. By using the black lipid membrane technique, it was shown that the semipurified metabolites (eluted from a SepPak C18 cartridge) of T. harzianum strain ES39 induced voltage-dependent conductivity. The high-performance liquid chromatography-purified metabolites of T. harzianum strain ES39 dissipated the mitochondrial membrane potential (Δψm) of human lung epithelial carcinoma cells (cell line A549). The semipurified metabolites (eluted from a SepPak C18 cartridge) of T. harzianum strain ES39 were analyzed by mass spectrometry (MS). Matrix-assisted laser desorption ionization and nanoflow electrospray ionization MS revealed five major peptaibols, each of which contained 18 residues and had a mass ranging from 1,719 to 1,775 Da. Their partial amino acid sequences were determined by collision-induced dissociation tandem MS. PMID:15294840

Optimization of solid-state fermentation conditions for Trichoderma harzianum using an orthogonal test.

PubMed

Zhang, J D; Yang, Q

2015-03-13

The aim of this study was to develop a protocol for the production of fungal bio-pesticides with high efficiency, low cost, and non-polluting fermentation, while also increasing their survival rate under field conditions. This is the first study to develop biocontrol Trichoderma harzianum transformants TS1 that are resistant to benzimidazole fungicides. Agricultural corn stover and wheat bran waste were used as a medium and inducing carbon source for solid fermentation. Spore production was observed, and the method was optimized using single-factor tests with 4 factors at 3 levels in an orthogonal experimental design to determine the optimal culture conditions for T. harzianum TS1. In this step, we determined the best conditions for fermenting the biocontrol fungi. The optimal culture conditions for T. harzianum TS1 were cultivated for 8 days, a ratio of straw to wheat bran of 1:3, ammonium persulfate as the nitrogen source, and a water content of 30 mL. Under optimal culture conditions, the sporulation of T. harzianum TS1 reached 1.49 x 10(10) CFU/g, which was 1.46-fold higher than that achieved before optimization. Increased sporulation of T. harzianum TS1 results in better utilization of space and nutrients to achieve control of plant pathogens. This method allows for the recycling of agricultural waste straw.

Comparative evaluation of two Trichoderma harzianum strains for major secondary metabolite production and antifungal activity.

PubMed

Ahluwalia, Vivek; Kumar, Jitendra; Rana, Virendra S; Sati, Om P; Walia, S

2015-01-01

This investigation was undertaken to identify the major secondary metabolite, produced by two Trichoderma harzianum strains (T-4 and T-5) with their antifungal activity against phytopathogenic fungi using poison food technique. The ethyl acetate extract was subjected to column chromatography using n-hexane, ethyl acetate and methanol gradually. Chromatographic separation of ethyl acetate extract of T. harzianum (T-4) resulted in the isolation and identification of palmitic acid (1), 1,8-dihydroxy-3-methylanthraquinone (2), 6-pentyl-2H-pyran-2-one (3), 2(5H)-furanone (4), stigmasterol (5) and β-sitosterol (6), while T. harzianum (T-5) gave palmitic acid (1), 1-hydroxy-3-methylanthraquinone (7), δ-decanolactone (8), 6-pentyl-2H-pyran-2-one (3), ergosterol (9), harzianopyridone (10) and 6-methyl-1,3,8-trihydroxyanthraquinone (11) as major metabolites. Among compounds screened for antifungal activity, compound 10 was found to be most active (EC50 35.9-50.2 μg mL(-1)). In conclusion, the present investigation provided significant information about antifungal activity and compounds isolated from two different strains of T. harzianum obtained from two different Himalayan locations.

Antagonism of Trichoderma harzianum ETS 323 on Botrytis cinerea mycelium in culture conditions.

PubMed

Cheng, Chi-Hua; Yang, Chia-Ann; Peng, Kou-Cheng

2012-11-01

ABSTRACT Previous studies have shown that the extracellular proteins of Trichoderma harzianum ETS 323 grown in the presence of deactivated Botrytis cinerea in culture include a putative l-amino acid oxidase and have suggested the involvement of this enzyme in the antagonistic mechanism. Here, we hypothesized that the mycoparasitic process of Trichoderma spp. against B. cinerea involves two steps; that is, an initial hyphal coiling stage and a subsequent hyphal coiling stage, with different coiling rates. The two-step antagonism of T. harzianum ETS 323 against B. cinerea during the mycoparasitic process in culture was evaluated using a biexponential equation. In addition, an l-amino acid oxidase (Th-l-AAO) was identified from T. harzianum ETS 323. The secretion of Th-l-AAO was increased when T. harzianum ETS 323 was grown with deactivated hyphae of B. cinerea. Moreover, in vitro assays indicated that Th-l-AAO effectively inhibited B. cinerea hyphal growth, caused cytosolic vacuolization in the hyphae, and led to hyphal lysis. Th-l-AAO also showed disease control against the development of B. cinerea on postharvest apple fruit and tobacco leaves. Furthermore, an apoptosis-like response, including the generation of reactive oxygen species, was observed in B. cinerea after treatment with Th-l-AAO, suggesting that Th-l-AAO triggers programmed cell death in B. cinerea. This may be associated with the two-step antagonism of T. harzianum ETS 323 against B. cinerea.

Cloning and sequencing of a cellobiohydrolase gene from Trichoderma harzianum FP108

Treesearch

Patrick Guilfoile; Ron Burns; Zu-Yi Gu; Matt Amundson; Fu-Hsian Chang

1999-01-01

A cbbl cellobiohydrolase gene was cloned and sequenced from the fungus Trichoderrna harzianum FP108. The cloning was performed by PCR amplification of T. harzianum genomic DNA, using PCR primers whose sequence was based on the cbbl gene from Tricboderma reesei. The 3' end of the gene was isolated by inverse...

Role of the Trichoderma harzianum Endochitinase Gene, ech42, in Mycoparasitism

PubMed Central

Carsolio, Carolina; Benhamou, Nicole; Haran, Shoshan; Cortés, Carlos; Gutiérrez, Ana; Chet, Ilan; Herrera-Estrella, Alfredo

1999-01-01

The role of the Trichoderma harzianum endochitinase (Ech42) in mycoparasitism was studied by genetically manipulating the gene that encodes Ech42, ech42. We constructed several transgenic T. harzianum strains carrying multiple copies of ech42 and the corresponding gene disruptants. The level of extracellular endochitinase activity when T. harzianum was grown under inducing conditions increased up to 42-fold in multicopy strains as compared with the wild type, whereas gene disruptants exhibited practically no activity. The densities of chitin labeling of Rhizoctonia solani cell walls, after interactions with gene disruptants were not statistically significantly different than the density of chitin labeling after interactions with the wild type. Finally, no major differences in the efficacies of the strains generated as biocontrol agents against R. solani or Sclerotium rolfsii were observed in greenhouse experiments. PMID:10049844

Involvement of the Transcriptional Coactivator ThMBF1 in the Biocontrol Activity of Trichoderma harzianum.

PubMed

Rubio, M Belén; Pardal, Alonso J; Cardoza, Rosa E; Gutiérrez, Santiago; Monte, Enrique; Hermosa, Rosa

2017-01-01

Trichoderma harzianum is a filamentous fungus well adapted to different ecological niches. Owing to its ability to antagonize a wide range of plant pathogens, it is used as a biological control agent in agriculture. Selected strains of T. harzianum are also able to increase the tolerance of plants to biotic and abiotic stresses. However, little is known about the regulatory elements of the T. harzianum transcriptional machinery and their role in the biocontrol by this species. We had previously reported the involvement of the transcription factor THCTF1 in the T. harzianum production of the secondary metabolite 6-pentyl-pyrone, an important volatile compound related to interspecies cross-talk. Here, we performed a subtractive hybridization to explore the genes regulated by THCTF1, allowing us to identify a multiprotein bridging factor 1 ( mbf1 ) homolog. The gene from T. harzianum T34 was isolated and characterized, and the generated Thmbf1 overexpressing transformants were used to investigate the role of this gene in the biocontrol abilities of the fungus against two plant pathogens. The transformants showed a reduced antifungal activity against Fusarium oxysporum f. sp. lycopersici race 2 (FO) and Botrytis cinerea (BC) in confrontation assays on discontinuous medium, indicating that the Thmbf1 gene could affect T. harzianum production of volatile organic compounds (VOC) with antifungal activity. Moreover, cellophane and dialysis membrane assays indicated that Thmbf1 overexpression affected the production of low molecular weight secreted compounds with antifungal activity against FO. Intriguingly, no correlation in the expression profiles, either in rich or minimal medium, was observed between Thmbf1 and the master regulator gene cross-pathway control ( cpc1 ). Greenhouse assays allowed us to evaluate the biocontrol potential of T. harzianum strains against BC and FO on susceptible tomato plants. The wild type strain T34 significantly reduced the necrotic leaf

Involvement of the Transcriptional Coactivator ThMBF1 in the Biocontrol Activity of Trichoderma harzianum

PubMed Central

Rubio, M. Belén; Pardal, Alonso J.; Cardoza, Rosa E.; Gutiérrez, Santiago; Monte, Enrique; Hermosa, Rosa

2017-01-01

Trichoderma harzianum is a filamentous fungus well adapted to different ecological niches. Owing to its ability to antagonize a wide range of plant pathogens, it is used as a biological control agent in agriculture. Selected strains of T. harzianum are also able to increase the tolerance of plants to biotic and abiotic stresses. However, little is known about the regulatory elements of the T. harzianum transcriptional machinery and their role in the biocontrol by this species. We had previously reported the involvement of the transcription factor THCTF1 in the T. harzianum production of the secondary metabolite 6-pentyl-pyrone, an important volatile compound related to interspecies cross-talk. Here, we performed a subtractive hybridization to explore the genes regulated by THCTF1, allowing us to identify a multiprotein bridging factor 1 (mbf1) homolog. The gene from T. harzianum T34 was isolated and characterized, and the generated Thmbf1 overexpressing transformants were used to investigate the role of this gene in the biocontrol abilities of the fungus against two plant pathogens. The transformants showed a reduced antifungal activity against Fusarium oxysporum f. sp. lycopersici race 2 (FO) and Botrytis cinerea (BC) in confrontation assays on discontinuous medium, indicating that the Thmbf1 gene could affect T. harzianum production of volatile organic compounds (VOC) with antifungal activity. Moreover, cellophane and dialysis membrane assays indicated that Thmbf1 overexpression affected the production of low molecular weight secreted compounds with antifungal activity against FO. Intriguingly, no correlation in the expression profiles, either in rich or minimal medium, was observed between Thmbf1 and the master regulator gene cross-pathway control (cpc1). Greenhouse assays allowed us to evaluate the biocontrol potential of T. harzianum strains against BC and FO on susceptible tomato plants. The wild type strain T34 significantly reduced the necrotic leaf lesions

New insights in Trichoderma harzianum antagonism of fungal plant pathogens by secreted protein analysis.

PubMed

Monteiro, Valdirene Neves; do Nascimento Silva, Roberto; Steindorff, Andrei Stecca; Costa, Fabio Teles; Noronha, Eliane Ferreira; Ricart, Carlos André Ornelas; de Sousa, Marcelo Valle; Vainstein, Marilene Henning; Ulhoa, Cirano José

2010-10-01

Trichoderma harzianum ALL42 were capable of overgrowing and degrading Rhizoctonia solani and Macrophomina phaseolina mycelia, coiling around the hyphae with formation of apressoria and hook-like structures. Hyphae of T. harzianum ALL42 did not show any coiling around Fusarium sp. hyphae suggesting that mycoparasitism may be different among the plant pathogens. In this study, a secretome analysis was used to identify some extracellular proteins secreted by T. harzianum ALL42 after growth on cell wall of M. phaseolina, Fusarium sp., and R. solani. The secreted proteins were analyzed by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. A total of 60 T. harzianum ALL42 secreted proteins excised from the gel were analyzed from the three growth conditions. While seven cell wall-induced proteins were identified, more than 53 proteins spots remain unidentified, indicating that these proteins are either novel proteins or proteins that have not yet been sequenced. Endochitinase, β-glucosidase, α-mannosidase, acid phosphatase, α-1,3-glucanase, and proteases were identified in the gel and also detected in the supernatant of culture.

Effects of Trichothecene Production on the Plant Defense Response and Fungal Physiology: Overexpression of the Trichoderma arundinaceum tri4 Gene in T. harzianum

PubMed Central

Cardoza, R. E.; McCormick, S. P.; Malmierca, M. G.; Olivera, E. R.; Alexander, N. J.; Monte, E.

2015-01-01

Trichothecenes are fungal sesquiterpenoid compounds, the majority of which have phytotoxic activity. They contaminate food and feed stocks, resulting in potential harm to animals and human beings. Trichoderma brevicompactum and T. arundinaceum produce trichodermin and harzianum A (HA), respectively, two trichothecenes that show different bioactive properties. Both compounds have remarkable antibiotic and cytotoxic activities, but in addition, trichodermin is highly phytotoxic, while HA lacks this activity when analyzed in vivo. Analysis of Fusarium trichothecene intermediates led to the conclusion that most of them, with the exception of the hydrocarbon precursor trichodiene (TD), have a detectable phytotoxic activity which is not directly related to the structural complexity of the intermediate. In the present work, the HA intermediate 12,13-epoxytrichothec-9-ene (EPT) was produced by expression of the T. arundinaceum tri4 gene in a transgenic T. harzianum strain that already produces TD after transformation with the T. arundinaceum tri5 gene. Purified EPT did not show antifungal or phytotoxic activity, while purified HA showed both antifungal and phytotoxic activities. However, the use of the transgenic T. harzianum tri4 strain induced a downregulation of defense-related genes in tomato plants and also downregulated plant genes involved in fungal root colonization. The production of EPT by the transgenic tri4 strain raised levels of erg1 expression and reduced squalene accumulation while not affecting levels of ergosterol. Together, these results indicate the complex interactions among trichothecene intermediates, fungal antagonists, and host plants. PMID:26150463

Liquid culture production of microsclerotia and submerged conidia by Trichoderma harzianum active against damping-off disease caused by Rhizoctonia solani.

PubMed

Kobori, Nilce N; Mascarin, Gabriel M; Jackson, Mark A; Schisler, David A

2015-04-01

Media and culturing protocols were identified that supported the formation of submerged conidia and microsclerotia (MS) by Trichoderma harzianum Rifai strain T-22 using liquid culture fermentation. Liquid media with a higher carbon concentration (36 g L(-1)) promoted MS formation at all C:N ratios tested. Hyphae aggregated to form MS after 2 d growth and after 7 d MS were fully melanized. This is the first report of MS formation by T. harzianum or any species of Trichoderma. Furthermore, submerged conidia formation was induced by liquid culture media, but yields, desiccation tolerance, and storage stability varied with C:N ratio and carbon rate. Air-dried MS granules (<4% moisture) retained excellent shelf life under cool and unrefrigerated storage conditions with no loss in conidial production. A low-cost complex nitrogen source based on cottonseed flour effectively supported high MS yields. Amending potting mix with dried MS formulations reduced or eliminated damping-off of melon seedlings caused by Rhizoctonia solani. Together, the results provide insights into the liquid culture production, stabilization process, and bioefficacy of the hitherto unreported MS of T. harzianum as a potential biofungicide for use in integrated management programs against soilborne diseases. Copyright © 2014 The British Mycological Society. All rights reserved.

Secretome analysis of the mycoparasitic fungus Trichoderma harzianum ALL 42 cultivated in different media supplemented with Fusarium solani cell wall or glucose.

PubMed

Ramada, Marcelo Henrique Soller; Steindorff, Andrei Stecca; Bloch, Carlos; Ulhoa, Cirano José

2016-02-01

Trichoderma harzianum is a fungus well known for its potential as a biocontrol agent against many fungal phytopathogens. The aim of this study was to characterize the proteins secreted by T. harzianum ALL42 when its spores were inoculated and incubated for 48 h in culture media supplemented with glucose (GLU) or with cell walls from Fusarium solani (FSCW), a phytopathogen that causes severe losses in common bean and soy crops in Brazil, as well as other crop diseases around the world. Trichoderma harzianum was able to grow in Trichoderma Liquid Enzyme Production medium (TLE) and Minimal medium (MM) supplemented with FSCW and in TLE+GLU, but was unable to grow in MM+GLU medium. Protein quantification showed that TLE+FSCW and MM+FSCW had 45- and 30- fold, respectively, higher protein concentration on supernatant when compared to TLE+GLU, and this difference was observable on 2D gel electrophoresis (2DE). A total of 94 out of 105 proteins excised from 2DE maps were identified. The only protein observed in all three conditions was epl1. In the media supplemented with FSCW, different hydrolases such as chitinases, β-1,3-glucanases, glucoamylases, α-1,3-glucanases and proteases were identified, along with other proteins with no known functions in mycoparasitism, such as npp1 and cys. Trichoderma harzianum showed a complex and diverse arsenal of proteins that are secreted in response to the presence of FSCW, with novel proteins not previously described in mycoparasitic-related studies. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

mRNA expression of EgCHI1, EgCHI2, and EgCHI3 in oil palm leaves (Elaeis guineesis Jacq.) after treatment with Ganoderma boninense pat. and Trichoderma harzianum Rifai.

PubMed

Naher, Laila; Tan, Soon Guan; Ho, Chai Ling; Yusuf, Umi Kalsom; Ahmad, Siti Hazar; Abdullah, Faridah

2012-01-01

Basal stem rot (BSR) disease caused by the fungus Ganoderma boninense is the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agent Trichoderma harzianum can protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensis Jacq.) chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated with G. boninense or T. harzianum alone or both. The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR. Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated with G. boninense alone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated with T. harzianum and G. boninense when compared to those of control and other treated plants. The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment.

The Cerato-Platanin protein Epl-1 from Trichoderma harzianum is involved in mycoparasitism, plant resistance induction and self cell wall protection

PubMed Central

Gomes, Eriston Vieira; Costa, Mariana do Nascimento; de Paula, Renato Graciano; Ricci de Azevedo, Rafael; da Silva, Francilene Lopes; Noronha, Eliane F.; José Ulhoa, Cirano; Neves Monteiro, Valdirene; Elena Cardoza, Rosa; Gutiérrez, Santiago; Nascimento Silva, Roberto

2015-01-01

Trichoderma harzianum species are well known as biocontrol agents against important fungal phytopathogens. Mycoparasitism is one of the strategies used by this fungus in the biocontrol process. In this work, we analyzed the effect of Epl-1 protein, previously described as plant resistance elicitor, in expression modulation of T. harzianum genes involved in mycoparasitism process against phytopathogenic fungi; self cell wall protection and recognition; host hyphae coiling and triggering expression of defense-related genes in beans plants. The results indicated that the absence of Epl-1 protein affects the expression of all mycoparasitism genes analyzed in direct confrontation assays against phytopathogen Sclerotinia sclerotiorum as well as T. harzianum itself; the host mycoparasitic coiling process and expression modulation of plant defense genes showing different pattern compared with wild type strain. These data indicated the involvement T. harzianum Epl-1 in self and host interaction and also recognition of T. harzianum as a symbiotic fungus by the bean plants. PMID:26647876

The Cerato-Platanin protein Epl-1 from Trichoderma harzianum is involved in mycoparasitism, plant resistance induction and self cell wall protection.

PubMed

Gomes, Eriston Vieira; Costa, Mariana do Nascimento; de Paula, Renato Graciano; de Azevedo, Rafael Ricci; da Silva, Francilene Lopes; Noronha, Eliane F; Ulhoa, Cirano José; Monteiro, Valdirene Neves; Cardoza, Rosa Elena; Gutiérrez, Santiago; Silva, Roberto Nascimento

2015-12-09

Trichoderma harzianum species are well known as biocontrol agents against important fungal phytopathogens. Mycoparasitism is one of the strategies used by this fungus in the biocontrol process. In this work, we analyzed the effect of Epl-1 protein, previously described as plant resistance elicitor, in expression modulation of T. harzianum genes involved in mycoparasitism process against phytopathogenic fungi; self cell wall protection and recognition; host hyphae coiling and triggering expression of defense-related genes in beans plants. The results indicated that the absence of Epl-1 protein affects the expression of all mycoparasitism genes analyzed in direct confrontation assays against phytopathogen Sclerotinia sclerotiorum as well as T. harzianum itself; the host mycoparasitic coiling process and expression modulation of plant defense genes showing different pattern compared with wild type strain. These data indicated the involvement T. harzianum Epl-1 in self and host interaction and also recognition of T. harzianum as a symbiotic fungus by the bean plants.

Transcriptome profile of Trichoderma harzianum IOC-3844 induced by sugarcane bagasse.

PubMed

Horta, Maria Augusta Crivelente; Vicentini, Renato; Delabona, Priscila da Silva; Laborda, Prianda; Crucello, Aline; Freitas, Sindélia; Kuroshu, Reginaldo Massanobu; Polikarpov, Igor; Pradella, José Geraldo da Cruz; Souza, Anete Pereira

2014-01-01

Profiling the transcriptome that underlies biomass degradation by the fungus Trichoderma harzianum allows the identification of gene sequences with potential application in enzymatic hydrolysis processing. In the present study, the transcriptome of T. harzianum IOC-3844 was analyzed using RNA-seq technology. The sequencing generated 14.7 Gbp for downstream analyses. De novo assembly resulted in 32,396 contigs, which were submitted for identification and classified according to their identities. This analysis allowed us to define a principal set of T. harzianum genes that are involved in the degradation of cellulose and hemicellulose and the accessory genes that are involved in the depolymerization of biomass. An additional analysis of expression levels identified a set of carbohydrate-active enzymes that are upregulated under different conditions. The present study provides valuable information for future studies on biomass degradation and contributes to a better understanding of the role of the genes that are involved in this process.

Effects of Trichothecene Production on the Plant Defense Response and Fungal Physiology: Overexpression of the Trichoderma arundinaceum tri4 Gene in T. harzianum.

PubMed

Cardoza, R E; McCormick, S P; Malmierca, M G; Olivera, E R; Alexander, N J; Monte, E; Gutiérrez, S

2015-09-01

Trichothecenes are fungal sesquiterpenoid compounds, the majority of which have phytotoxic activity. They contaminate food and feed stocks, resulting in potential harm to animals and human beings. Trichoderma brevicompactum and T. arundinaceum produce trichodermin and harzianum A (HA), respectively, two trichothecenes that show different bioactive properties. Both compounds have remarkable antibiotic and cytotoxic activities, but in addition, trichodermin is highly phytotoxic, while HA lacks this activity when analyzed in vivo. Analysis of Fusarium trichothecene intermediates led to the conclusion that most of them, with the exception of the hydrocarbon precursor trichodiene (TD), have a detectable phytotoxic activity which is not directly related to the structural complexity of the intermediate. In the present work, the HA intermediate 12,13-epoxytrichothec-9-ene (EPT) was produced by expression of the T. arundinaceum tri4 gene in a transgenic T. harzianum strain that already produces TD after transformation with the T. arundinaceum tri5 gene. Purified EPT did not show antifungal or phytotoxic activity, while purified HA showed both antifungal and phytotoxic activities. However, the use of the transgenic T. harzianum tri4 strain induced a downregulation of defense-related genes in tomato plants and also downregulated plant genes involved in fungal root colonization. The production of EPT by the transgenic tri4 strain raised levels of erg1 expression and reduced squalene accumulation while not affecting levels of ergosterol. Together, these results indicate the complex interactions among trichothecene intermediates, fungal antagonists, and host plants. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum.

PubMed

Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Coelho, Alexandre Siqueira Guedes; Miller, Robert Neil Gerard; Pappas, Georgios Joannis; Ulhoa, Cirano José; Noronha, Eliane Ferreira

2014-03-18

The species of T. harzianum are well known for their biocontrol activity against plant pathogens. However, few studies have been conducted to further our understanding of its role as a biological control agent against S. sclerotiorum, a pathogen involved in several crop diseases around the world. In this study, we have used RNA-seq and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum gene expression during growth on cell wall of S. sclerotiorum (SSCW) or glucose. RT-qPCR was also used to examine genes potentially involved in biocontrol, during confrontation between T. harzianum and S. sclerotiorum. Data obtained from six RNA-seq libraries were aligned onto the T. harzianum CBS 226.95 reference genome and compared after annotation using the Blast2GO suite. A total of 297 differentially expressed genes were found in mycelia grown for 12, 24 and 36 h under the two different conditions: supplemented with glucose or SSCW. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on SSCW or glucose. We identified various genes of biotechnological value encoding proteins with functions such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. To validate the expression profile, RT-qPCR was performed using 20 randomly chosen genes. RT-qPCR expression profiles were in complete agreement with the RNA-Seq data for 17 of the genes evaluated. The other three showed differences at one or two growth times. During the confrontation assay, some genes were up-regulated during and after contact, as shown in the presence of SSCW which is commonly used as a model to mimic this interaction. The present study is the first initiative to use RNA-seq for identification of differentially expressed genes in T. harzianum strain TR274, in response to the phytopathogenic fungus S. sclerotiorum. It provides insights into the mechanisms of

Effect of trichothecene production on the plant defense response and fungal physiology: overexpression of Trichoderma arundinaceum tri4 gene in T. harzianum

USDA-ARS?s Scientific Manuscript database

Trichothecenes are fungal sesquiterpenoid compounds, the majority of which have phytotoxic activity. They contaminate food and feed stocks, resulting in potential harm to animals and human beings. Trichoderma brevicompactum and T. arundinaceum produce trichodermin and harzianum A (HA), respectively,...

[Characterization of a carbendazim-degrading Trichoderma sp. T2-2 and its application in bioremediation].

PubMed

Tian, Liansheng; Chen, Fei

2009-07-01

To obtain carbendazim-degrading microbial strains, and to use them for bioremediation of contaminated soil. A carbendazim-degrading bacterium T2-2 was isolated from the screening of drug-tolerated mutants Trichoderma strains. High-pressure liquid chromatography-mass spectrometry (HPLC-MS) analysis showed the presence of the metabolites after shake incubation of the Trichoderma T2-2 at temperature 25 degrees C, 200 r/min in mineral salt medium that contained 100 mg/ L carbendazim. We prepared T2-2 bioremediation agents from crop straw through solid fermentation. By inoculating T2-2 in soil, we performed a bioremediation test of sterilized soil and original soil at 0.1 mg/g dry soil of carbendazim concentration and 10(7) cfu/g dry soil of inoculating amount. In addition, we also conducted a control effect experiment of T2-2 against fusarium wilt of cucumber. The metabolites detected by HPLC-MS were 2-aminobenzimidazole, benzimidazole, and 2-aminobenxinitrile in the culture filtrate after 2 days of incubation. Carbendazim and metabolites could no longer be detected through the High-pressure liquid chromatography (HPLC) analysis in the culture filtrate after 5 days of incubation. In the soil bioremediation test, carbendazim in the sterilized soil was degraded completely after 6 days of inoculation, whereas the process only needed 4 days in original soil. It showed crop straw could function as co-metabolic substrate and promote co-metabolism of T2-2 and indigenous microorganisms. Moreover, the efficiency of T2-2 against cucumber fusarium wilt might reach 81.7%, which is superior to chemical pesticide. T2-2 could degrade carbendazim in soil and thus control plant disease.

Identification of differentially expressed genes from Trichoderma harzianum during growth on cell wall of Fusarium solani as a tool for biotechnological application.

PubMed

Vieira, Pabline Marinho; Coelho, Alexandre Siqueira Guedes; Steindorff, Andrei Stecca; de Siqueira, Saulo José Linhares; Silva, Roberto do Nascimento; Ulhoa, Cirano José

2013-03-15

The species of T. harzianum are well known for their biocontrol activity against many plant pathogens. However, there is a lack of studies concerning its use as a biological control agent against F. solani, a pathogen involved in several crop diseases. In this study, we have used subtractive library hybridization (SSH) and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum genes expression during growth on cell wall of F. solani (FSCW) or glucose. RT-qPCR was also used to examine the regulation of 18 genes, potentially involved in biocontrol, during confrontation between T. harzianum and F. solani. Data obtained from two subtractive libraries were compared after annotation using the Blast2GO suite. A total of 417 and 78 readable EST sequence were annotated in the FSCW and glucose libraries, respectively. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on FSCW or glucose. We identified various genes of biotechnological value encoding to proteins which function such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. Fifteen genes were up-regulated and sixteen were down-regulated at least at one-time point during growth of T. harzianum in FSCW. During the confrontation assay most of the genes were up-regulated, mainly after contact, when the interaction has been established. This study demonstrates that T. harzianum expressed different genes when grown on FSCW compared to glucose. It provides insights into the mechanisms of gene expression involved in mycoparasitism of T. harzianum against F. solani. The identification and evaluation of these genes may contribute to the development of an efficient biological control agent.

The Trichoderma harzianum demon: complex speciation history resulting in coexistence of hypothetical biological species, recent agamospecies and numerous relict lineages

PubMed Central

2010-01-01

Background The mitosporic fungus Trichoderma harzianum (Hypocrea, Ascomycota, Hypocreales, Hypocreaceae) is an ubiquitous species in the environment with some strains commercially exploited for the biological control of plant pathogenic fungi. Although T. harzianum is asexual (or anamorphic), its sexual stage (or teleomorph) has been described as Hypocrea lixii. Since recombination would be an important issue for the efficacy of an agent of the biological control in the field, we investigated the phylogenetic structure of the species. Results Using DNA sequence data from three unlinked loci for each of 93 strains collected worldwide, we detected a complex speciation process revealing overlapping reproductively isolated biological species, recent agamospecies and numerous relict lineages with unresolved phylogenetic positions. Genealogical concordance and recombination analyses confirm the existence of two genetically isolated agamospecies including T. harzianum sensu stricto and two hypothetical holomorphic species related to but different from H. lixii. The exact phylogenetic position of the majority of strains was not resolved and therefore attributed to a diverse network of recombining strains conventionally called 'pseudoharzianum matrix'. Since H. lixii and T. harzianum are evidently genetically isolated, the anamorph - teleomorph combination comprising H. lixii/T. harzianum in one holomorph must be rejected in favor of two separate species. Conclusions Our data illustrate a complex speciation within H. lixii - T. harzianum species group, which is based on coexistence and interaction of organisms with different evolutionary histories and on the absence of strict genetic borders between them. PMID:20359347

Cadmium induces cadmium-tolerant gene expression in the filamentous fungus Trichoderma harzianum.

PubMed

Cacciola, Santa O; Puglisi, Ivana; Faedda, Roberto; Sanzaro, Vincenzo; Pane, Antonella; Lo Piero, Angela R; Evoli, Maria; Petrone, Goffredo

2015-11-01

The filamentous fungus Trichoderma harzianum, strain IMI 393899, was able to grow in the presence of the heavy metals cadmium and mercury. The main objective of this research was to study the molecular mechanisms underlying the tolerance of the fungus T. harzianum to cadmium. The suppression subtractive hybridization (SSH) method was used for the characterization of the genes of T. harzianum implicated in cadmium tolerance compared with those expressed in the response to the stress induced by mercury. Finally, the effects of cadmium exposure were also validated by measuring the expression levels of the putative genes coding for a glucose transporter, a plasma membrane ATPase, a Cd(2+)/Zn(2+) transporter protein and a two-component system sensor histidine kinase YcbA, by real-time-PCR. By using the aforementioned SSH strategy, it was possible to identify 108 differentially expressed genes of the strain IMI 393899 of T. harzianum grown in a mineral substrate with the addition of cadmium. The expressed sequence tags identified by SSH technique were encoding different genes that may be involved in different biological processes, including those associated to primary and secondary metabolism, intracellular transport, transcription factors, cell defence, signal transduction, DNA metabolism, cell growth and protein synthesis. Finally, the results show that in the mechanism of tolerance to cadmium a possible signal transduction pathway could activate a Cd(2+)/Zn(2+) transporter protein and/or a plasma membrane ATPase that could be involved in the compartmentalization of cadmium inside the cell.

Mycoparasitism studies of Trichoderma harzianum against Sclerotinia sclerotiorum: evaluation of antagonism and expression of cell wall-degrading enzymes genes.

PubMed

Troian, Rogério Fraga; Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Arruda, Walquiria; Ulhoa, Cirano José

2014-10-01

Trichoderma spp. are known for their biocontrol activity against several plant pathogens. A specific isolate of Trichoderma harzianum, 303/02, has the potential to inhibit the growth of Sclerotinia sclerotiorum, an important agent involved in several crop diseases. In this study, the interaction between T. harzianum 303/02 and mycelia, sclerotia and apothecia of S. sclerotiorum was studied by scanning electron microscopy. RT-qPCR was used to examine the expression of 11 genes potentially involved in biocontrol. T. harzianum 303/02 parasitizes S. sclerotiorum by forming branches that coil around the hyphae. The fungus multiplied abundantly at the sclerotia and apothecia surface, forming a dense mycelium that penetrated the inner surface of these structures. The levels of gene expression varied according to the type of structure with which T. harzianum was interacting. The data also showed the presence of synergistic action between the cell-wall degrading enzymes.

mRNA Expression of EgCHI1, EgCHI2, and EgCHI3 in Oil Palm Leaves (Elaeis guineesis Jacq.) after Treatment with Ganoderma boninense Pat. and Trichoderma harzianum Rifai

PubMed Central

Naher, Laila; Tan, Soon Guan; Ho, Chai Ling; Yusuf, Umi Kalsom; Ahmad, Siti Hazar; Abdullah, Faridah

2012-01-01

Background. Basal stem rot (BSR) disease caused by the fungus Ganoderma boninense is the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agent Trichoderma harzianum can protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensis Jacq.) chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated with G. boninense or T. harzianum alone or both. Methods. The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR. Results. Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated with G. boninense alone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated with T. harzianum and G. boninense when compared to those of control and other treated plants. Conclusion. The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment. PMID:22919345

Identification of differentially expressed genes from Trichoderma harzianum during growth on cell wall of Fusarium solani as a tool for biotechnological application

PubMed Central

2013-01-01

Background The species of T. harzianum are well known for their biocontrol activity against many plant pathogens. However, there is a lack of studies concerning its use as a biological control agent against F. solani, a pathogen involved in several crop diseases. In this study, we have used subtractive library hybridization (SSH) and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum genes expression during growth on cell wall of F. solani (FSCW) or glucose. RT-qPCR was also used to examine the regulation of 18 genes, potentially involved in biocontrol, during confrontation between T. harzianum and F. solani. Results Data obtained from two subtractive libraries were compared after annotation using the Blast2GO suite. A total of 417 and 78 readable EST sequence were annotated in the FSCW and glucose libraries, respectively. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on FSCW or glucose. We identified various genes of biotechnological value encoding to proteins which function such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. Fifteen genes were up-regulated and sixteen were down-regulated at least at one-time point during growth of T. harzianum in FSCW. During the confrontation assay most of the genes were up-regulated, mainly after contact, when the interaction has been established. Conclusions This study demonstrates that T. harzianum expressed different genes when grown on FSCW compared to glucose. It provides insights into the mechanisms of gene expression involved in mycoparasitism of T. harzianum against F. solani. The identification and evaluation of these genes may contribute to the development of an efficient biological control agent. PMID:23497274

Inoculation of tomato seedlings with Trichoderma Harzianum and Arbuscular Mycorrhizal Fungi and their effect on growth and control of wilt in tomato seedlings.

PubMed

Mwangi, Margaret W; Monda, Ethel O; Okoth, Sheila A; Jefwa, Joyce M

2011-04-01

A green house study was conducted to investigate the ability of an isolate of Trichoderma harzianum (P52) and arbuscular mycorrhizal fungi (AMF) in enhancing growth and control of a wilt pathogen caused by Fusarium oxysporum f. sp. lycopersici in tomato seedlings. The plants were grown in plastic pots filled with sterilized soils. There were four treatments applied as follows; P52, AMF, AMF + P52 and a control. A completely randomized design was used and growth measurements and disease assessment taken after 3, 6 and 9 weeks. Treatments that significantly (P < 0.05) enhanced heights and root dry weights were P52, AMF and a treatment with a combination of both P52 and AMF when compared the control. The treatment with both P52 and AMF significantly (P < 0.05) enhanced all growth parameters (heights; shoot and root dry weight) investigated compared to the control. Disease severity was generally lower in tomato plants grown with isolate P52 and AMF fungi either individually or when combined together, though the effect was not statistically significant (P≥ 0.05). A treatment combination of P52 + AMF had less trend of severity as compared to each individual fungus. T. harzianum and AMF can be used to enhance growth in tomato seedlings.

Systemic Resistance to Powdery Mildew in Brassica napus (AACC) and Raphanus alboglabra (RRCC) by Trichoderma harzianum TH12

PubMed Central

Alkooranee, Jawadayn Talib; Yin, Yongtai; Aledan, Tamarah Raad; Jiang, Yingfen; Lu, Guangyuan; Wu, Jiangsheng; Li, Maoteng

2015-01-01

Trichoderma harzianum TH12 is a microbial pesticide for certain rapeseed diseases. The mechanism of systemic resistance induced by TH12 or its cell-free culture filtrate (CF) in Brassica napus (AACC) and Raphanus alboglabra (RRCC) to powdery mildew disease caused by ascomycete Erysiphe cruciferarum was investigated. In this study, we conducted the first large-scale global study on the cellular and molecular aspects of B. napus and R. alboglabra infected with E. cruciferarum. The histological study showed the resistance of R. alboglabra to powdery mildew disease. The growth of fungal colonies was not observed on R. alboglabra leaves at 1, 2, 4, 6, 8, and 10 days post-inoculation (dpi), whereas this was clearly observed on B. napus leaves after 6 dpi. In addition, the gene expression of six plant defense-related genes, namely, PR-1, PR-2 (a marker for SA signaling), PR-3, PDF 1.2 (a marker for JA/ET signaling), CHI620, and CHI570, for both genotypes were analyzed in the leaves of B. napus and R. alboglabra after treatment with TH12 or CF and compared with the non-treated ones. The qRT-PCR results showed that the PR-1 and PR-2 expression levels increased in E. cruciferarum-infected leaves, but decreased in the TH12-treated leaves compared with leaves treated with CF. The expression levels of PR-3 and PDF1.2 decreased in plants infected by E. cruciferarum. However, expression levels increased when the leaves were treated with TH12. For the first time, we disclosed the nature of gene expression in B. napus and R. alboglabra to explore the resistance pathways in the leaves of both genotypes infected and non-infected by powdery mildew and inoculated or non-inoculated with elicitor factors. Results suggested that R. alboglabra exhibited resistance to powdery mildew disease, and the application of T. harzianum and its CF are a useful tool to facilitate new protection methods for resist or susceptible plants. PMID:26540161

Systemic Resistance to Powdery Mildew in Brassica napus (AACC) and Raphanus alboglabra (RRCC) by Trichoderma harzianum TH12.

PubMed

Alkooranee, Jawadayn Talib; Yin, Yongtai; Aledan, Tamarah Raad; Jiang, Yingfen; Lu, Guangyuan; Wu, Jiangsheng; Li, Maoteng

2015-01-01

Trichoderma harzianum TH12 is a microbial pesticide for certain rapeseed diseases. The mechanism of systemic resistance induced by TH12 or its cell-free culture filtrate (CF) in Brassica napus (AACC) and Raphanus alboglabra (RRCC) to powdery mildew disease caused by ascomycete Erysiphe cruciferarum was investigated. In this study, we conducted the first large-scale global study on the cellular and molecular aspects of B. napus and R. alboglabra infected with E. cruciferarum. The histological study showed the resistance of R. alboglabra to powdery mildew disease. The growth of fungal colonies was not observed on R. alboglabra leaves at 1, 2, 4, 6, 8, and 10 days post-inoculation (dpi), whereas this was clearly observed on B. napus leaves after 6 dpi. In addition, the gene expression of six plant defense-related genes, namely, PR-1, PR-2 (a marker for SA signaling), PR-3, PDF 1.2 (a marker for JA/ET signaling), CHI620, and CHI570, for both genotypes were analyzed in the leaves of B. napus and R. alboglabra after treatment with TH12 or CF and compared with the non-treated ones. The qRT-PCR results showed that the PR-1 and PR-2 expression levels increased in E. cruciferarum-infected leaves, but decreased in the TH12-treated leaves compared with leaves treated with CF. The expression levels of PR-3 and PDF1.2 decreased in plants infected by E. cruciferarum. However, expression levels increased when the leaves were treated with TH12. For the first time, we disclosed the nature of gene expression in B. napus and R. alboglabra to explore the resistance pathways in the leaves of both genotypes infected and non-infected by powdery mildew and inoculated or non-inoculated with elicitor factors. Results suggested that R. alboglabra exhibited resistance to powdery mildew disease, and the application of T. harzianum and its CF are a useful tool to facilitate new protection methods for resist or susceptible plants.

Saprotrophic and Mycoparasitic Components of Aggressiveness of Trichoderma harzianum Groups toward the Commercial Mushroom Agaricus bisporus

PubMed Central

Williams, Josie; Clarkson, John M.; Mills, Peter R.; Cooper, Richard M.

2003-01-01

We examined the mycoparasitic and saprotrophic behavior of isolates representing groups of Trichoderma harzianum to establish a mechanism for the aggressiveness towards Agaricus bisporus in infested commercial compost. Mycoparasitic structures were infrequently observed in interaction zones on various media, including compost, with cryoscanning electron microscopy. T. harzianum grows prolifically in compost in the absence or presence of A. bisporus, and the aggressive European (Th2) and North American (Th4) isolates produced significantly higher biomasses (6.8- and 7.5-fold, respectively) in compost than did nonaggressive, group 1 isolates. All groups secreted depolymerases that could attack the cell walls of A. bisporus and of wheat straw, and some were linked to aggressiveness. Growth on mushroom cell walls in vitro resulted in rapid production of chymoelastase and trypsin-like proteases by only the Th2 and Th4 isolates. These isolates also produced a dominant protease isoform (pI 6.22) and additional chitinase isoforms. On wheat straw, Th4 produced distinct isoforms of cellulase and laminarinase, but there was no consistent association between levels or isoforms of depolymerases and aggressiveness. Th3's distinctive profiles confirmed its reclassification as Trichoderma atroviride. Proteases and glycanases were detected for the first time in sterilized compost colonized by T. harzianum. Xylanase dominated, and some isoforms were unique to compost, as were some laminarinases. We hypothesize that aggressiveness results from competition, antagonism, or parasitism but only as a component of, or following, extensive saprotrophic growth involving degradation of wheat straw cell walls. PMID:12839799

Genome Sequencing and Comparative Analysis of the Biocontrol Agent Trichoderma harzianum sensu stricto TR274

DOE Office of Scientific and Technical Information (OSTI.GOV)

Steindorff, Andrei S.; Noronha, Elilane F.; Ulhoa, Cirano J.

2015-03-17

Biological control is a complex process which requires many mechanisms and a high diversity of biochemical pathways. The species of Trichoderma harzianum are well known for their biocontrol activity against many plant pathogens. To gain new insights into the biocontrol mechanism used by T. harzianum, we sequenced the isolate TR274 genome using Illumina. The assembly was performed using AllPaths-LG with a maximum coverage of 100x. The assembly resulted in 2282 contigs with a N50 of 37033bp. The genome size generated was 40.8 Mb and the GC content was 47.7%, similar to other Trichoderma genomes. Using the JGI Annotation Pipeline wemore » predicted 13,932 genes with a high transcriptome support. CEGMA tests suggested 100% genome completeness and 97.9% of RNA-SEQ reads were mapped to the genome. The phylogenetic comparison using orthologous proteins with all Trichoderma genomes sequenced at JGI, corroborates the Trichoderma (T. asperellum and T. atroviride), Longibrachiatum (T. reesei and T. longibrachiatum) and Pachibasium (T. harzianum and T. virens) section division described previously. The comparison between two Trichoderma harzianum species suggests a high genome similarity but some strain-specific expansions. Analyses of the secondary metabolites, CAZymes, transporters, proteases, transcription factors were performed. The Pachybasium section expanded virtually all categories analyzed compared with the other sections, specially Longibrachiatum section, that shows a clear contraction. These results suggests that these proteins families have an important role in their respective phenotypes. Future analysis will improve the understanding of this complex genus and give some insights about its lifestyle and the interactions with the environment.« less

Thctf1 transcription factor of Trichoderma harzianum is involved in 6-pentyl-2H-pyran-2-one production and antifungal activity.

PubMed

Rubio, M Belén; Hermosa, Rosa; Reino, José Luis; Collado, Isidro G; Monte, Enrique

2009-01-01

We describe the cloning and characterization of the Trichoderma harzianum Thctf1 gene, which shows high sequence identity with a transcription factor gene of Fusarium solani f. sp. pisi. In T. harzianum, disruption of the Thctf1 gene by homologous recombination gave rise to transformants that in plate experiments did not show the yellow pigmentation observed in the wild-type strain. In several Trichoderma spp. a yellow pigmentation and a coconut aroma have been related to the production of 6-pentyl-2H-pyran-2-one (6PP) compounds. Prompted by this, we explored whether the loss of pigmentation in the Thctf1 null mutants of T. harzianum might be related to the synthesis of 6PP. Chromatographic and spectroscopic analyses revealed that the disruptants did not produce two secondary metabolites, derived from 6PP and not previously described in the Trichoderma genus, that are present in wild-type culture filtrates. Since 6PP is a recognized antifungal compound, this ability was analyzed in both the disruptants and wild-type, observing that the Thctf1 null mutants of T. harzianum had reduced antimicrobial capacity. Our results point to the significant role of THCTF1 in the production of secondary metabolites and in the antifungal activity of T. harzianum.

Understanding the cellulolytic system of Trichoderma harzianum P49P11 and enhancing saccharification of pretreated sugarcane bagasse by supplementation with pectinase and α-L-arabinofuranosidase.

PubMed

Delabona, Priscila da Silva; Cota, Júnio; Hoffmam, Zaira Bruna; Paixão, Douglas Antonio Alvaredo; Farinas, Cristiane Sanchez; Cairo, João Paulo Lourenço Franco; Lima, Deise Juliana; Squina, Fábio Marcio; Ruller, Roberto; Pradella, José Geraldo da Cruz

2013-03-01

Supplementation of cellulase cocktails with accessory enzymes can contribute to a higher hydrolytic capacity in releasing fermentable sugars from plant biomass. This study investigated which enzymes were complementary to the enzyme set of Trichoderma harzianum in the degradation of sugarcane bagasse. Specific activities of T. harzianum extract on different substrates were compared with the extracts of Penicillium echinulatum and Trichoderma reesei, and two commercial cellulase preparations. Complementary analysis of the secretome of T. harzianum was also used to identify which enzymes were produced during growth on pretreated sugarcane bagasse. These analyses enabled the selection of the enzymes pectinase and α-L-arabinofuranosidase (AF) to be further investigated as supplements to the T. harzianum extract. The effect of enzyme supplementation on the efficiency of sugarcane bagasse saccharification was evaluated using response surface methodology. The supplementation of T. harzianum enzymatic extract with pectinase and AF increased the efficiency of hydrolysis by up to 116%. Copyright © 2012 Elsevier Ltd. All rights reserved.

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.

PubMed

Mercado, José A; Barceló, Marta; Pliego, Clara; Rey, Manuel; Caballero, José L; Muñoz-Blanco, Juan; Ruano-Rosa, David; López-Herrera, Carlos; de Los Santos, Berta; Romero-Muñoz, Fernando; Pliego-Alfaro, Fernando

2015-12-01

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.

Impact of salicylic acid- and jasmonic acid-regulated defences on root colonization by Trichoderma harzianum T-78.

PubMed

Martínez-Medina, Ainhoa; Appels, Freek V W; van Wees, Saskia C M

2017-08-03

We recently found that the beneficial fungus Trichoderma harzianum T-78 primes tomato plants for salicylic acid (SA)- and jasmonic acid (JA)-regulated defenses, resulting in enhanced resistance against the root knot nematode Meloidogyne incognita. By using SA- and JA-impaired mutant lines and exogenous hormonal application, here we investigated whether the SA- and JA-pathways also have a role in T-78 root colonization of Arabidopsis thaliana. Endophytic colonization by T-78 was faster in the SA-impaired mutant sid2 than in the wild type. Moreover, elicitation of SA-dependent defenses by SA application reduced T-78 colonization, indicating that the SA-pathway affects T-78 endophytism. In contrast, elicitation of the JA-pathway, which antagonized SA-dependent defenses, resulted in enhanced endophytic colonization by T-78. These findings are in line with our previous observation that SA-dependent defenses are repressed by T-78, which likely aids colonization by the endophytic fungus.

Molecular characterization of intergeneric hybrid between Aspergillus oryzae and Trichoderma harzianum by protoplast fusion.

PubMed

Patil, N S; Patil, S M; Govindwar, S P; Jadhav, J P

2015-02-01

Protoplast fusion between Aspergillus oryzae and Trichoderma harzianum and application of fusant in degradation of shellfish waste. The filamentous chitinolytic fungal strains A. oryzae NCIM 1272 and T. harzianum NCIM 1185 were selected as parents for protoplast fusion. Viable protoplasts were released from fungal mycelium using enzyme cocktail containing 5 mg ml(-1) lysing enzymes from T. harzianum, 0.06 mg ml(-1) β-glucuronidase from Helix pomatia and 1 mg ml(-1) purified Penicillium ochrochloron chitinase in 0.8 mol l(-1) sorbitol as an osmotic stabilizer. Intergeneric protoplast fusion was carried out using 60% polyethylene glycol as a fusogen. At optimum conditions, the regeneration frequency of the fused protoplasts on colloidal chitin medium and fusion frequency were calculated. Fusant showed higher rate of growth pattern, chitinase activity and protein content than parents. Fusant formation was confirmed by morphological markers, viz. colony morphology and spore size and denaturation gradient gel electrophoresis (DGGE). This study revealed protoplast fusion between A. oryzae and T. harzianum significantly enhanced chitinase activity which ultimately provides potential strain for degradation of shellfish waste. Consistency in the molecular characterization results using DGGE is the major outcome of this study which can be emerged as a fundamental step in fusant identification. Now it is need to provide attention over effective chitin degradation to manage shrimp processing issues. In this aspect, ability of fusant to degrade shellfish waste efficiently in short incubation time revealed discovery of potential strain in the reclamation of seafood processing crustacean bio-waste. © 2014 The Society for Applied Microbiology.

Isolation and functional analysis of Thmfs1, the first major facilitator superfamily transporter from the biocontrol fungus Trichoderma harzianum.

PubMed

Liu, Mu; Liu, Jun; Wang, Wei Min

2012-10-01

A novel major facilitator superfamily (MFS) transporter gene, Thmfs1, was isolated from Trichoderma harzianum (T. harzianum). A Thmfs1 over-expressing mutant displayed enhanced antifungal activity and fungicide tolerance, while the Thmfs1 disruption mutant showed the opposite trend. Trichodermin production in Thmfs1 disruption group (185 mg l(-1)) was decreased by less than 17 % compared to the parental strain, suggesting that Thmfs1 is not mainly responsible for trichodermin secretion. Real-time PCR showed that Thmfs1 transcript level could be induced by a certain range of trichodermin concentrations, while expression of Tri5, encoding a trichodiene synthase, was strongly inhibited under these conditions. To our knowledge, Thmfs1 is the first MFS transporter gene identified in T. harzianum.

Induction of SA-signaling pathway and ethylene biosynthesis in Trichoderma harzianum-treated tomato plants after infection of the root-knot nematode Meloidogyne incognita.

PubMed

Leonetti, Paola; Zonno, Maria Chiara; Molinari, Sergio; Altomare, Claudio

2017-04-01

Salicylic acid-signaling pathway and ethylene biosynthesis were induced in tomato treated with Trichoderma harzianum when infected by root-knot nematodes and limited the infection by activation of SAR and ethylene production. Soil pre-treatment with Trichoderma harzianum (Th) strains ITEM 908 (T908) and T908-5 decreased susceptibility of tomato to Meloidogyne incognita, as assessed by restriction in nematode reproduction and development. The effect of T. harzianum treatments on plant defense was detected by monitoring the expression of the genes PR-1/PR-5 and JERF3/ACO, markers of the SA- and JA/ET-dependent signaling pathways, respectively. The compatible nematode-plant interaction in absence of fungi caused a marked suppression of PR-1, PR-5, and ACO gene expressions, either locally or systemically, whilst expression of JERF3 gene resulted unaffected. Conversely, when plants were pre-treated with Th-strains, over-expression of PR-1, PR-5, and ACO genes was observed in roots 5 days after nematode inoculation. JERF3 gene expression did not change in Th-colonized plants challenged with nematodes. In the absence of nematodes, Trichoderma-root interaction was characterized by the inhibition of both SA-dependent signaling pathway and ET biosynthesis, and, in the case of PR-1 and ACO genes, this inhibition was systemic. JERF3 gene expression was systemically restricted only at the very early stages of plant-fungi interaction. Data presented indicate that Th-colonization primed roots for Systemic Acquired Resistance (SAR) against root-knot nematodes and reacted to nematode infection more efficiently than untreated plants. Such a response probably involves also activation of ET production, through an augmented transcription of the ACO gene, which encodes for the enzyme catalyzing the last step of ET biosynthesis. JA signaling and Induced Systemic Resistance (ISR) do not seem to be involved in the biocontrol action of the tested Th-strains against RKNs.

Analysis of genomic regions of Trichoderma harzianum IOC-3844 related to biomass degradation.

PubMed

Crucello, Aline; Sforça, Danilo Augusto; Horta, Maria Augusta Crivelente; dos Santos, Clelton Aparecido; Viana, Américo José Carvalho; Beloti, Lilian Luzia; de Toledo, Marcelo Augusto Szymanski; Vincentz, Michel; Kuroshu, Reginaldo Massanobu; de Souza, Anete Pereira

2015-01-01

Trichoderma harzianum IOC-3844 secretes high levels of cellulolytic-active enzymes and is therefore a promising strain for use in biotechnological applications in second-generation bioethanol production. However, the T. harzianum biomass degradation mechanism has not been well explored at the genetic level. The present work investigates six genomic regions (~150 kbp each) in this fungus that are enriched with genes related to biomass conversion. A BAC library consisting of 5,760 clones was constructed, with an average insert length of 90 kbp. The assembled BAC sequences revealed 232 predicted genes, 31.5% of which were related to catabolic pathways, including those involved in biomass degradation. An expression profile analysis based on RNA-Seq data demonstrated that putative regulatory elements, such as membrane transport proteins and transcription factors, are located in the same genomic regions as genes related to carbohydrate metabolism and exhibit similar expression profiles. Thus, we demonstrate a rapid and efficient tool that focuses on specific genomic regions by combining a BAC library with transcriptomic data. This is the first BAC-based structural genomic study of the cellulolytic fungus T. harzianum, and its findings provide new perspectives regarding the use of this species in biomass degradation processes.

Biocontrol potential of Trichoderma harzianum isolate T-aloe against Sclerotinia sclerotiorum in soybean.

PubMed

Zhang, Fuli; Ge, Honglian; Zhang, Fan; Guo, Ning; Wang, Yucheng; Chen, Long; Ji, Xiue; Li, Chengwei

2016-03-01

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum (Lib.) de Bary is a major disease of soybean (Glycine max (L.) Merr.). At present, we revealed the three-way interaction between Trichoderma harzianum T-aloe, pathogen S. sclerotiorum and soybean plants in order to demonstrate biocontrol mechanism and evaluate biocontrol potential of T-aloe against S. sclerotiorum in soybean. In our experiments, T-aloe inhibited the growth of S. sclerotiorum with an efficiency of 56.3% in dual culture tests. T-aloe hyphae grew in parallel or intertwined with S. sclerotiorum hyphae and produced hooked contact branches, indicating mycoparasitism. Plate tests showed that T-aloe culture filtrate inhibited S. sclerotiorum growth with an inhibition efficiency of 51.2% and sclerotia production. T-aloe pretreatment showed growth-promoting effect on soybean plants. The activities of peroxidase, superoxide dismutase, and catalase increased, and the hydrogen peroxide (H2O2) as well as the superoxide radical (O2(-)) content in soybean leaves decreased after T-aloe pretreatment in response to S. sclerotiorum pathogen challenge. T-aloe treatment diminished damage caused by pathogen stress on soybean leaf cell membrane, and increased chlorophyll as well as total phenol contents. The defense-related genes PR1, PR2, and PR3 were expressed in the leaves of T-aloe-treated plants. In summary, T-aloe displayed biocontrol potential against S. sclerotiorum. This is the first report of unraveling biocontrol potential of Trichoderma Spp. to soybean sclerotinia stem rot from the three-way interaction between the biocontrol agent, pathogen S. sclerotiorum and soybean plants. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Abiotic stresses affect Trichoderma harzianum T39-induced resistance to downy mildew in grapevine.

PubMed

Roatti, Benedetta; Perazzolli, Michele; Gessler, Cesare; Pertot, Ilaria

2013-12-01

Enhancement of plant defense through the application of resistance inducers seems a promising alternative to chemical fungicides for controlling crop diseases but the efficacy can be affected by abiotic factors in the field. Plants respond to abiotic stresses with hormonal signals that may interfere with the mechanisms of induced systemic resistance (ISR) to pathogens. In this study, we exposed grapevines to heat, drought, or both to investigate the effects of abiotic stresses on grapevine resistance induced by Trichoderma harzianum T39 (T39) to downy mildew. Whereas the efficacy of T39-induced resistance was not affected by exposure to heat or drought, it was significantly reduced by combined abiotic stresses. Decrease of leaf water potential and upregulation of heat-stress markers confirmed that plants reacted to abiotic stresses. Basal expression of defense-related genes and their upregulation during T39-induced resistance were attenuated by abiotic stresses, in agreement with the reduced efficacy of T39. The evidence reported here suggests that exposure of crops to abiotic stress should be carefully considered to optimize the use of resistance inducers, especially in view of future global climate changes. Expression analysis of ISR marker genes could be helpful to identify when plants are responding to abiotic stresses, in order to optimize treatments with resistance inducers in field.

Elucidation of biocontrol mechanisms of Trichoderma harzianum against different plant fungal pathogens: Universal yet host specific response.

PubMed

Sharma, Vivek; Salwan, Richa; Sharma, Prem N; Kanwar, S S

2017-02-01

In the present study, different transcripts of Trichoderma harzianum ThHP-3 were evaluated for their response against four fungal pathogens Fusarium oxysporum, Colletotrichum capsici, Colletotrichum truncatum and Gloesercospora sorghi using RT-qPCR. The time course study of T. harzianum transcripts related to signal transduction, lytic enzymes, secondary metabolites and various transporters revealed variation in expression against four fungal pathogens. In a broader term, the transcripts were upregulated at various time intervals but the optimum expression of cyp3, abc, nrp, tga1, pmk, ech42 and glh20 varied with respect to host fungi. Additionally, the expression of transcripts related to transporters/cytochromes was also observed against Fusarium oxysporum after 96h whereas transcripts related to secondary metabolites and lytic enzymes showed significant difference in expression against Colletotrichum spp. from 72 to 96h. This is first study on transcriptomic response of T. harzianum against pathogenic fungi which shows their host specific response. Copyright © 2016 Elsevier B.V. All rights reserved.

A novel L-amino acid oxidase from Trichoderma harzianum ETS 323 associated with antagonism of Rhizoctonia solani.

PubMed

Yang, Chia-Ann; Cheng, Chi-Hua; Lo, Chaur-Tsuen; Liu, Shu-Ying; Lee, Jeng-Woei; Peng, Kou-Cheng

2011-05-11

Trichoderma spp. are used as biocontrol agents against phytopathogens such as Rhizoctonia solani, but their biocontrol mechanisms are poorly understood. A novel L-amino oxidase (Th-LAAO) was identified from the extracellular proteins of Trichoderma harzianum ETS 323. Here, we show a FAD-binding glycoprotein with the best substrate specificity constant for L-phenylalanine. Although the amino acid sequence of Th-LAAO revealed limited homology (16-24%) to other LAAO members, a highly conserved FAD-binding motif was identified in the N-terminus. Th-LAAO was shown to be a homodimeric protein, but the monomeric form was predominant when grown in the presence of deactivated Rhizoctonia solani. Furthermore, in vitro assays demonstrated that Th-LAAO had an antagonistic effect against Rhizoctonia solani and a stimulatory one on hyphal density and sporulation in T. harzianum ETS 323. These findings further our understanding of T. harzianum as a biocontrol agent and provide insight into the biological function of l-amino acid oxidase.

Analysis of Genomic Regions of Trichoderma harzianum IOC-3844 Related to Biomass Degradation

PubMed Central

Crucello, Aline; Sforça, Danilo Augusto; Horta, Maria Augusta Crivelente; dos Santos, Clelton Aparecido; Viana, Américo José Carvalho; Beloti, Lilian Luzia; de Toledo, Marcelo Augusto Szymanski; Vincentz, Michel; Kuroshu, Reginaldo Massanobu; de Souza, Anete Pereira

2015-01-01

Trichoderma harzianum IOC-3844 secretes high levels of cellulolytic-active enzymes and is therefore a promising strain for use in biotechnological applications in second-generation bioethanol production. However, the T. harzianum biomass degradation mechanism has not been well explored at the genetic level. The present work investigates six genomic regions (~150 kbp each) in this fungus that are enriched with genes related to biomass conversion. A BAC library consisting of 5,760 clones was constructed, with an average insert length of 90 kbp. The assembled BAC sequences revealed 232 predicted genes, 31.5% of which were related to catabolic pathways, including those involved in biomass degradation. An expression profile analysis based on RNA-Seq data demonstrated that putative regulatory elements, such as membrane transport proteins and transcription factors, are located in the same genomic regions as genes related to carbohydrate metabolism and exhibit similar expression profiles. Thus, we demonstrate a rapid and efficient tool that focuses on specific genomic regions by combining a BAC library with transcriptomic data. This is the first BAC-based structural genomic study of the cellulolytic fungus T. harzianum, and its findings provide new perspectives regarding the use of this species in biomass degradation processes. PMID:25836973

Characterization of the cellulolytic secretome of Trichoderma harzianum during growth on sugarcane bagasse and analysis of the activity boosting effects of swollenin.

PubMed

A L Rocha, Vanessa; N Maeda, Roberto; Pereira, Nei; F Kern, Marcelo; Elias, Luisa; Simister, Rachael; Steele-King, Clare; Gómez, Leonardo D; McQueen-Mason, Simon J

2016-03-01

This study demonstrates the production of an active enzyme cocktail produced by growing Trichoderma harzianum on sugarcane bagasse. The component enzymes were identified by LCMS-MS. Glycosyl hydrolases were the most abundant class of proteins, representing 67% of total secreted protein. Other carbohydrate active enzymes involved in cell wall deconstruction included lytic polysaccharide mono-oxygenases (AA9), carbohydrate-binding modules, carbohydrate esterases and swollenin, all present at levels of 1%. In total, proteases and lipases represented 5 and 1% of the total secretome, respectively, with the rest of the secretome being made up of proteins of unknown or putative function. This enzyme cocktail was efficient in catalysing the hydrolysis of sugarcane bagasse cellulolignin to fermentable sugars for potential use in ethanol production. Apart from mapping the secretome of T. harzianum, which is a very important tool to understand the catalytic performance of enzyme cocktails, the gene coding for T. harzianum swollenin was expressed in Aspergillus niger. This novel aspect in this work, allowed increasing the swollenin concentration by 95 fold. This is the first report about the heterologous expression of swollenin from T. harzianum, and the findings are of interest in enriching enzyme cocktail with this important accessory protein which takes part in the cellulose amorphogenesis. Despite lacking detectable glycoside activity, the addition of swollenin of T. harzianum increased by two-fold the hydrolysis efficiency of a commercial cellulase cocktail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:327-336, 2016. © 2016 American Institute of Chemical Engineers.

Microencapsuling aerial conidia of Trichoderma harzianum through spray drying at elevated temperatures

USDA-ARS?s Scientific Manuscript database

Trichoderma conidia are mostly produced by solid fermentation systems. Inoculum is produced by liquid culturing, and then transferred to solid substrate for aerial conidial production. Aerial conidia of T. harzianum are hydrophilic in nature, and it is difficult to separate them from the solid subst...

Analysis of Phaseolus vulgaris Response to Its Association with Trichoderma harzianum (ALL-42) in the Presence or Absence of the Phytopathogenic Fungi Rhizoctonia solani and Fusarium solani

PubMed Central

Pereira, Jackeline L.; Queiroz, Rayner M. L.; Charneau, Sébastien O.; Felix, Carlos R.; Ricart, Carlos A. O.; da Silva, Francilene Lopes; Steindorff, Andrei Stecca; Ulhoa, Cirano J.; Noronha, Eliane F.

2014-01-01

The present study was carried out to evaluate the ability of Trichoderma harzianum (ALL 42-isolated from Brazilian Cerrado soil) to promote common bean growth and to modulate its metabolism and defense response in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani using a proteomic approach. T. harzianum was able to promote common bean plants growth as shown by the increase in root/foliar areas and by size in comparison to plants grown in its absence. The interaction was shown to modulate the expression of defense-related genes (Glu1, pod3 and lox1) in roots of P. vulgaris. Proteomic maps constructed using roots and leaves of plants challenged or unchallenged by T. harzianum and phytopathogenic fungi showed differences. Reference gels presented differences in spot distribution (absence/presence) and relative volumes of common spots (up or down-regulation). Differential spots were identified by peptide fingerprinting MALDI-TOF mass spectrometry. A total of 48 identified spots (19 for leaves and 29 for roots) were grouped into protein functional classes. For leaves, 33%, 22% and 11% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively. For roots, 17.2%, 24.1% and 10.3% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively. PMID:24878929

Analysis of Phaseolus vulgaris response to its association with Trichoderma harzianum (ALL-42) in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani.

PubMed

Pereira, Jackeline L; Queiroz, Rayner M L; Charneau, Sébastien O; Felix, Carlos R; Ricart, Carlos A O; da Silva, Francilene Lopes; Steindorff, Andrei Stecca; Ulhoa, Cirano J; Noronha, Eliane F

2014-01-01

The present study was carried out to evaluate the ability of Trichoderma harzianum (ALL 42-isolated from Brazilian Cerrado soil) to promote common bean growth and to modulate its metabolism and defense response in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani using a proteomic approach. T. harzianum was able to promote common bean plants growth as shown by the increase in root/foliar areas and by size in comparison to plants grown in its absence. The interaction was shown to modulate the expression of defense-related genes (Glu1, pod3 and lox1) in roots of P. vulgaris. Proteomic maps constructed using roots and leaves of plants challenged or unchallenged by T. harzianum and phytopathogenic fungi showed differences. Reference gels presented differences in spot distribution (absence/presence) and relative volumes of common spots (up or down-regulation). Differential spots were identified by peptide fingerprinting MALDI-TOF mass spectrometry. A total of 48 identified spots (19 for leaves and 29 for roots) were grouped into protein functional classes. For leaves, 33%, 22% and 11% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively. For roots, 17.2%, 24.1% and 10.3% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively.

Thc6 protein, isolated from Trichoderma harzianum, can induce maize defense response against Curvularia lunata.

PubMed

Fan, Lili; Fu, Kehe; Yu, Chuanjin; Li, Yingying; Li, Yaqian; Chen, Jie

2015-05-01

Mutant T66 was isolated from 450 mutants (constructed with Agrobacterium tumefaciens-mediated transformation method) of Trichoderma harzianum. Maize seeds coated with T66 were more susceptible to Curvularia lunata when compared with those coated with wild-type (WT) strain. The disease index of maize treated with T66 and WT were 62.5 and 42.1%, respectively. Further research showed T-DNA has inserted into the ORF of one gene, which resulted in the functional difference between WT and T66. The gene was cloned and named Thc6, which encodes a novel 327 amino acid protein. To investigate its function, we obtained knockout, complementation, and overexpression mutants of Thc6. Challenge inoculation studies suggested that the Thc6 overexpression mutant can reduce the disease index of maize inbred line Huangzao 4 against the leaf spot pathogen (C. lunata). Meanwhile, The Thc6 mutants were found to affect the resistance of maize inbred line Huangzao 4 against C. lunata by enhancing the activation of jasmonate-responsive genes expression. Liquid chromatography-mass spectrometry (LC-MS) data further confirmed that the concentration of jasmonate in the induced maize exhibits a parallel change tendency with the expression level of defense-related genes. Hence, the Thc6 gene could be participated in the induced resistance of maize inbred line Huangzao 4 against C. lunata infection through a jasmonic acid-dependent pathway. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Mechanism of triphenylmethane Cresol Red degradation by Trichoderma harzianum M06.

PubMed

Nor, Nurafifah Mohd; Hadibarata, Tony; Zubir, Meor Mohd Fikri Ahmad; Lazim, Zainab Mat; Adnan, Liyana Amalina; Fulazzaky, Mohamad Ali

2015-11-01

Cresol Red belongs to the triphenylmethane (TPM) class of dyes which are potentially carcinogenic or mutagenic. However, very few studies on biodegradation of Cresol Red were investigated as compared to other type dyes such as azo and anthraquinone dye. The aim of this work is to evaluate triphenylmethane dye Cresol Red degradation by fungal strain isolated from the decayed wood in Johor Bahru, Malaysia. Detailed taxonomic studies identified the organisms as Trichoderma species and designated as strain Trichoderma harzianum M06. In this study, Cresol Red was decolorized up to 88% within 30 days under agitation condition by Trichoderma harzianum M06. Data analysis revealed that a pH value of 3 yielded a highest degradation rate among pH concentrations (73%), salinity concentrations of 100 g/L (73%), and a volume of 0.1 mL of Tween 80 (79%). Induction in the enzyme activities of manganese peroxidase, lignin peroxidase, laccase, 1,2- and 2,3-dioxygenase indicates their involvement in Cresol Red removal. Various analytical studies such as Thin-Layer Chromatography (TLC), UV-Vis spectrophotometer, and Gas chromatography mass spectrometry (GC-MS) confirmed the biotransformation of Cresol Red by the fungus. Two metabolites were identified in the treated medium: 2,4-dihydroxybenzoic acid (t R 7.3 min and m/z 355) and 2-hydroxybenzoic acid (t R 8.6 min and m/z 267). Based on these products, a probable pathway has been proposed for the degradation of Cresol Red by Trichoderma harzianum M06.

The potentiality of Trichoderma harzianum in alleviation the adverse effects of salinity in faba bean plants.

PubMed

Abd El-Baki, G K; Mostafa, Doaa

2014-12-01

The interaction between sodium chloride and Trichoderma harzianum (T24) on growth parameters, ion contents, MDA content, proline, soluble proteins as well as SDS page protein profile were studied in Vicia faba Giza 429. A sharp reduction was found in fresh and dry mass of shoots and roots with increasing salinity. Trichoderma treatments promoted the growth criteria as compared with corresponding salinized plants. The water content and leaf area exhibited a marked decrease with increasing salinity. Trichoderma treatments induced a progressive increase in both parameters. Both proline and MDA contents were increased progressively as the salinity rose in the soil. Trichoderma treatments considerably retarded the accumulation of both parameters in shoots and roots. Both Na+ and K+ concentration increased in both organs by enhancing salinity levels. The treatment with Trichoderma harzianum enhanced the accumulation of both ions. Exposure of plants to different concentrations of salinity, or others treated with Trichoderma harzianum produced marked changes in their protein pattern. Three types of alterations were observed: the synthesis of certain proteins declined significantly, specific synthesis of certain other proteins were markedly observed and synthesis of a set specific protein was induced de novo in plant treated with Trichoderma harzianum.

Trichoderma harzianum strain SQR-T37 and its bio-organic fertilizer could control Rhizoctonia solani damping-off disease in cucumber seedlings mainly by the mycoparasitism.

PubMed

Huang, Xinqi; Chen, Lihua; Ran, Wei; Shen, Qirong; Yang, Xingming

2011-08-01

Damping-off disease is caused by Rhizoctonia solani and leads to serious loss in many crops. Biological control is an efficient and environmentally friendly way to prevent damping-off disease. Optical micrographs, scanning electron micrographs, and the determination of hydrolytic enzymes were used to investigate the antagonism of Trichoderma harzianum SQR-T37 (SQR-T37) against R. solani. Experiments were performed in pots to assess the in vivo disease-control efficiency of SQR-T37 and bio-organic fertilizer. The results indicate that the mycoparasitism was the main mechanism accounting for the antagonistic activity of SQR-T37. In one experiment, the population of R. solani was decreased from 10(6) internal transcribed spacer (ITS) copies per gram soil to 10(4) ITS copies per gram soil by the presence of the antagonist. In this experiment, 45% of the control efficiency was obtained when 8 g of SQR-T37 hyphae per gram soil was applied. In a second experiment, as much as 81.82% of the control efficiency was obtained when bio-organic fertilizer (SQR-T37 fermented organic fertilizer, BIO) was applied compared to only 27.27% of the control efficiency when only 4 g of SQR-T37 hyphae per gram soil was applied. Twenty days after incubation, the population of T. harzianum was 4.12 × 10(7) ITS copies per gram soil in the BIO treatment, which was much higher than that in the previous treatment (8.77 × 10(5) ITS copies per gram soil), where only SQR-T37 was applied. The results indicated that SQR-T37 was a potent antagonist against R. solani in a mycoparasitic way that decreased the population of the pathogen. Applying BIO was more efficient than SQR-T37 application alone because it stabilized the population of the antagonist.

Trichoderma harzianum containing 1-aminocyclopropane-1-carboxylate deaminase and chitinase improved growth and diminished adverse effect caused by Fusarium oxysporum in soybean.

PubMed

Zhang, Fuli; Chen, Can; Zhang, Fan; Gao, Lidong; Liu, Jidong; Chen, Long; Fan, Xiaoning; Liu, Chang; Zhang, Ke; He, Yuting; Chen, Chen; Ji, Xiue

2017-03-01

An isolate, named Trichoderma harzianum T-soybean, showed growth-promoting for soybean seedlings and induced resistance to Fusarium oxysporum under greenhouse. Compared to control soybean seedlings, fresh weight, dry weight, lateral root number, chlorophyll content, root activity and soluble protein of plants pretreated with T-soybean increased, but initial pod height reduced. Furthermore, we found that T-soybean inhibited the growth of F. oxysporum by parasitic function. In addition, plate test results showed that culture filtrates of T-soybean also inhibited significantly F. oxysporum growth. Meanwhile, T-soybean treatment obviously reduced disease severity and induced quickly the H 2 O 2 and O 2 - burst as well as pathogenesis related protein gene (PR3) expression after F. oxysporum inoculation, and subsequently diminished the cell damage in soybean caused by the pathogen challenge. Reactive oxygen species (ROS) scavenging enzymes activity analysis showed that the activities of peroxidase (POD), polyphenol oxidase (PPO) and superoxide dismutase (SOD) increased significantly in T-soybean pretreated plants. These results suggested that T-soybean treatment induced resistance in soybean seedlings to F. oxysporum by companying the production of ROS and the increasing of ROS scavenging enzymes activity as well as PR3 expression. Copyright © 2016 Elsevier GmbH. All rights reserved.

Antagonistic studies and hyphal interactions of the new antagonist Aspergillus piperis against some phytopathogenic fungi in vitro in comparison with Trichoderma harzianum.

PubMed

El-Debaiky, Samah A

2017-12-01

The present study represents, for the first time, the detailed studies about the hyphal interactions of Aspergillus piperis, as a new antagonist, against some isolated plant pathogenic fungi (Alternaria alternata, Alternaria solani, Botrytis cinerea, Sclerotium cepivorum and Sclerotinia sclerotiorum) in vitro. The bio-controlling capability of A. piperis against the tested phytopathogens was tested using the dual culture method. This experiment revealed that A. piperis had antagonistic activity and reduced the growth of the tested phytopathogens and grew over their mycelia in the paired plates. Also, several antagonistic mechanisms were recorded, in this study, between A. piperis and the tested phytopathogens using the microscopic examination. The bio-controlling activity and the antagonistic mechanisms exhibited by the new antagonist, A. piperis were compared with those obtained by the common antagonist, Trichoderma harzianum against the same phytopathogens. The obtained results showed that, A. piperis was more effective than T. harzianum in inhibiting all the tested species in the dual culture plates. The best result was 81.85% inhibition percentage against S. sclerotiorum by A. piperis while, T. harzianum exhibits only 45.18%. Moreover, several antagonistic mechanisms and hyphal interactions were investigated among the hyphae of both A.piperis and T. harzianum and the hyphae of the tested phytopathogens. These mechanisms were summarized as; mycoparasitism (coiling and penetration of the hyphae) and antibiosis in the form of lysis of the hyphal cells and spores, denaturation and breaking of the hyphae. The indirect interaction (antibiosis) and the direct mycoparasitism were observed by A. piperis against all the tested phytopathogens, but it attacked the hyphae and conidiophores of A. alternata by only the antibiosis interaction. The microscopic examination revealed also that T. harzianum attacked the tested phytopathogens by both antibiosis and mycoparasitism

An Antifungal Exo-α-1,3-Glucanase (AGN13.1) from the Biocontrol Fungus Trichoderma harzianum

PubMed Central

Ait-Lahsen, Hassane; Soler, Andrés; Rey, Manuel; de la Cruz, Jesús; Monte, Enrique; Llobell, Antonio

2001-01-01

Trichoderma harzianum secretes α-1,3-glucanases when it is grown on polysaccharides, fungal cell walls, or autoclaved mycelium as a carbon source (simulated antagonistic conditions). We have purified and characterized one of these enzymes, named AGN13.1. The enzyme was monomeric and slightly basic. AGN13.1 was an exo-type α-1,3-glucanase and showed lytic and antifungal activity against fungal plant pathogens. Northern and Western analyses indicated that AGN13.1 is induced by conditions that simulated antagonism. We propose that AGN13.1 contributes to the antagonistic response of T. harzianum. PMID:11722942

Generation, annotation and analysis of ESTs from Trichoderma harzianum CECT 2413

PubMed Central

Vizcaíno, Juan Antonio; González, Francisco Javier; Suárez, M Belén; Redondo, José; Heinrich, Julian; Delgado-Jarana, Jesús; Hermosa, Rosa; Gutiérrez, Santiago; Monte, Enrique; Llobell, Antonio; Rey, Manuel

2006-01-01

Background The filamentous fungus Trichoderma harzianum is used as biological control agent of several plant-pathogenic fungi. In order to study the genome of this fungus, a functional genomics project called "TrichoEST" was developed to give insights into genes involved in biological control activities using an approach based on the generation of expressed sequence tags (ESTs). Results Eight different cDNA libraries from T. harzianum strain CECT 2413 were constructed. Different growth conditions involving mainly different nutrient conditions and/or stresses were used. We here present the analysis of the 8,710 ESTs generated. A total of 3,478 unique sequences were identified of which 81.4% had sequence similarity with GenBank entries, using the BLASTX algorithm. Using the Gene Ontology hierarchy, we performed the annotation of 51.1% of the unique sequences and compared its distribution among the gene libraries. Additionally, the InterProScan algorithm was used in order to further characterize the sequences. The identification of the putatively secreted proteins was also carried out. Later, based on the EST abundance, we examined the highly expressed genes and a hydrophobin was identified as the gene expressed at the highest level. We compared our collection of ESTs with the previous collections obtained from Trichoderma species and we also compared our sequence set with different complete eukaryotic genomes from several animals, plants and fungi. Accordingly, the presence of similar sequences in different kingdoms was also studied. Conclusion This EST collection and its annotation provide a significant resource for basic and applied research on T. harzianum, a fungus with a high biotechnological interest. PMID:16872539

Fusarium oxysporum induces the production of proteins and volatile organic compounds by Trichoderma harzianum T-E5.

PubMed

Zhang, Fengge; Yang, Xingming; Ran, Wei; Shen, Qirong

2014-10-01

Trichoderma species have been used widely as biocontrol agents for the suppression of soil-borne pathogens. However, some antagonistic mechanisms of Trichoderma are not well characterized. In this study, a series of laboratory experiments were designed to characterize the importance of mycoparasitism, exoenzymes, and volatile organic compounds (VOCs) by Trichoderma harzianum T-E5 for the control of Fusarium oxysporum f. sp. cucumerinum (FOC). We further tested whether these mechanisms were inducible and upregulated in presence of FOC. The results were as follows: T-E5 heavily parasitized FOC by coiling and twisting the entire mycelium of the pathogen in dual cultures. T-E5 growing medium conditioned with deactivated FOC (T2) showed more proteins and higher cell wall-degrading enzyme activities than T1, suggesting that FOC could induce the upregulation of exoenzymes. The presence of deactivated FOC (T2') also resulted in the upregulation of VOCs that five and eight different types T-E5-derived VOCs were identified from T1' and T2', respectively. Further, the excreted VOCs in T2' showed significantly higher antifungal activities against FOC than T1'. In conclusion, mycoparasitism of T-E5 against FOC involved mycelium contact and the production of complex extracellular substances. Together, these data provide clues to help further clarify the interactions between these fungi. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Novel mutations in β-tubulin gene in Trichoderma harzianum mutants resistant to methyl benzimidazol-2-yl carbamate.

PubMed

Li, M; Zhang, H Y; Liang, B

2013-01-01

Twelve-low resistant (LR) mutants of Trichoderma harzianum with the capability of grow fast at 0.8 μg/mL methyl benzimidazol-2-yl carbamate (MBC) were obtained using UV mutagenesis. MR and HR mutants which could grow fast at 10 and 100 μg/mL MBC, respectively, were isolated by step-up selection protocols in which UV-treated mutants were induced and mycelial sector screening was made in plates with growth medium. Subsequently, β-tubulin genes of 14 mutants were cloned to describe-the molecular lesion likely to be responsible-for MBC resistance. Comparison of the β-tubulin sequences of the mutant and sensitive strains of T. harzianum revealed 2 new MBC-binding sites differed from those in other plant pathogens. A single mutation at-amino acid 168, having Phe (TTC) instead of Ser (TCC)', was demonstrated for the HR mutant; a double mutation in amino acid 13 resulting in the substitution of Gly (GGC) by Val (GTG) was observed in β-tubulin gene of MR mutant. On the other hand, no substitutions were identified in the β-tubulin gene and its 5'-flanking regions in 12 LR mutants of T. harzianum.

Oxidative damage induced by heat stress could be relieved by nitric oxide in Trichoderma harzianum LTR-2.

PubMed

Yu, Yang; Yang, Zijun; Guo, Kai; Li, Zhe; Zhou, Hongzi; Wei, Yanli; Li, Jishun; Zhang, Xinjian; Harvey, Paul; Yang, Hetong

2015-04-01

Trichoderma harzianum is an important commercial biocontrol fungal agent. The temperature has been shown to be an important parameter and strain-specific to the mycelia growth of fungi, but less report makes the known of the mechanisms in T. harzianum. In our study, a 6-h treatment of heat increased the thiobarbituric acid reactive substances (TBARS) and nitric oxide (NO) concentration in mycelia to 212 and 230 % the level of the control, respectively. The exogenous NO donor sodium nitroprusside (150 μM) reduced the TBARS concentration to 53 % of that under heat stress (HS). At the same time, the NO-specific scavenger at 250 μM, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-1-oxyl-3-oxide, prevented the exogenous NO-relieved TBARS accumulation under HS. The increased NO concentration under HS was reduced 41 % by the NO synthase (NOS) inhibitor L-N(G)-nitroarginine methyl ester, but not the nitrate reductase (NR) inhibitor tungstate. Our study exhibited that NO can protect the mycelia of T. harzianum from HS and reduce the oxidative damage by enhancing the activity of NOS and NR.

Secretomic survey of Trichoderma harzianum grown on plant biomass substrates.

PubMed

Gómez-Mendoza, Diana Paola; Junqueira, Magno; do Vale, Luis Henrique Ferreira; Domont, Gilberto Barbosa; Ferreira Filho, Edivaldo Ximenes; Sousa, Marcelo Valle de; Ricart, Carlos André Ornelas

2014-04-04

The present work aims at characterizing T. harzianum secretome when the fungus is grown in synthetic medium supplemented with one of the four substrates: glucose, cellulose, xylan, and sugarcane bagasse (SB). The characterization was done by enzymatic assays and proteomic analysis using 2-DE/MALDI-TOF and gel-free shotgun LC-MS/MS. The results showed that SB induced the highest cellulolytic and xylanolytic activities when compared with the other substrates, while remarkable differences in terms of number and distribution of protein spots in 2-DE gels were also observed among the samples. Additionally, treatment of the secretomes with PNGase F revealed that most spot trails in 2-DE gels corresponded to N-glycosylated proteoforms. The LC-MS/MS analysis of the samples identified 626 different protein groups, including carbohydrate-active enzymes and accessory, noncatalytic, and cell-wall-associated proteins. Although the SB-induced secretome displayed the highest cellulolytic and xylanolytic activities, it did not correspond to a higher proteome complexity because CM-cellulose-induced secretome was significantly more diverse. Among the identified proteins, 73% were exclusive to one condition, while only 5% were present in all samples. Therefore, this study disclosed the variation of T. harzianum secretome in response to different substrates and revealed the diversity of the fungus enzymatic toolbox.

The Variations of Glycolysis and TCA Cycle Intermediate Levels Grown in Iron and Copper Mediums of Trichoderma harzianum.

PubMed

Tavsan, Zehra; Ayar Kayali, Hulya

2015-05-01

The efficiency of optimal metabolic function by microorganism depends on various parameters, especially essential metal supplementation. In the present study, the effects of iron and copper metals on metabolism were investigated by determination of glycolysis and tricarboxylic acid (TCA) cycle metabolites' levels with respect to the metal concentrations and incubation period in Trichoderma harzianum. The pyruvate and citrate levels of T. harzianum increased up to 15 mg/L of copper via redirection of carbon flux though glycolysis by suppression of pentose phosphate pathway (PPP). However, the α-ketoglutarate levels decreased at concentration higher than 5 mg/L of copper to overcome damage of oxidative stress. The fumarate levels correlated with the α-ketoglutarate levels because of substrate limitation. Besides, in T. harzianum cells grown in various concentrations of iron-containing medium, the intracellular pyruvate, citrate, and α-ketoglutarate levels showed positive correlation with iron concentration due to modifying of expression of glycolysis and TCA cycle enzymes via a mechanism involving cofactor or allosteric regulation. However, as a result of consuming of prior substrates required for fumarate production, its levels rose up to 10 mg/L.

Preparation, characterisation and viability of encapsulated Trichoderma harzianum UPM40 in alginate-montmorillonite clay.

PubMed

Adzmi, Fariz; Meon, Sariah; Musa, Mohamed Hanafi; Yusuf, Nor Azah

2012-01-01

Microencapsulation is a process by which tiny parcels of an active ingredient are packaged within a second material for the purpose of shielding the active ingredient from the surrounding environment. This study aims to determine the ability of the microencapsulation technique to improve the viability of Trichoderma harzianum UPM40 originally isolated from healthy groundnut roots as effective biological control agents (BCAs). Alginate was used as the carrier for controlled release, and montmorillonite clay (MMT) served as the filler. The encapsulated Ca-alginate-MMT beads were characterised using Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and scanning electron microscopy (SEM). The FTIR results showed the interaction between the functional groups of alginate and MMT in the Ca-alginate-MMT beads. Peaks at 1595, 1420 and 1020 cm(-1) characterised alginate, and peaks at 1028 and 453 cm(-1) characterised MMT; both sets of peaks appeared in the Ca-alginate-MMT FTIR spectrum. The TGA analysis showed an improvement in the thermal stability of the Ca-alginate-MMT beads compared with the alginate beads alone. SEM analysis revealed a homogeneous distribution of the MMT particles throughout the alginate matrix. T. harzianum UPM40 was successfully encapsulated in the Ca-alginate-MMT beads. Storage analysis of the encapsulated T. harzianum UPM40 showed that the low storage temperature of 5°C resulted in significantly (p < 0.05) better storage compared with room temperature (30°C).

Cellulase from Trichoderma harzianum interacts with roots and triggers induced systemic resistance to foliar disease in maize

PubMed Central

Saravanakumar, Kandasamy; Fan, Lili; Fu, Kehe; Yu, Chuanjin; Wang, Meng; Xia, Hai; Sun, Jianan; Li, Yaqian; Chen, Jie

2016-01-01

Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize. PMID:27830829

Cellulase from Trichoderma harzianum interacts with roots and triggers induced systemic resistance to foliar disease in maize.

PubMed

Saravanakumar, Kandasamy; Fan, Lili; Fu, Kehe; Yu, Chuanjin; Wang, Meng; Xia, Hai; Sun, Jianan; Li, Yaqian; Chen, Jie

2016-11-10

Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize.

Fungal Biomass Protein Production from Trichoderma harzianum Using Rice Polishing

PubMed Central

Mustafa, Ghulam; Arshad, Muhammad

2017-01-01

Industrially important enzymes and microbial biomass proteins have been produced from fungi for more than 50 years. High levels of crude protein as much as 45% are present in fungal biomass with balanced essential amino acids. The aim of this study was to access the potential of Trichoderma harzianum to produce fungal biomass protein from rice polishings. Maximum biomass yield was obtained at 5% (w/v) rice polishings after 72 h of incubation at 28°C at pH 4. Carbon and nitrogen ratio of 20 : 1 gave significantly higher production of fungal biomass protein. The FBP in the 75 L fermenter contained 49.50% crude protein, 32.00% true protein, 19.45% crude fiber, 9.62% ash, 11.5% cellulose content, and 0.325% RNA content. The profile of amino acids of final FBP exhibited that all essential amino acids were present in great quantities. The FBP produced by this fungus has been shown to be of good nutritional value for supplementation to poultry. The results presented in this study have practical implications in that the fungus T. harzianum could be used successfully to produce fungal biomass protein using rice polishings. PMID:28367444

Fungal Biomass Protein Production from Trichoderma harzianum Using Rice Polishing.

PubMed

Ahmed, Sibtain; Mustafa, Ghulam; Arshad, Muhammad; Rajoka, Muhammad Ibrahim

2017-01-01

Industrially important enzymes and microbial biomass proteins have been produced from fungi for more than 50 years. High levels of crude protein as much as 45% are present in fungal biomass with balanced essential amino acids. The aim of this study was to access the potential of Trichoderma harzianum to produce fungal biomass protein from rice polishings. Maximum biomass yield was obtained at 5% (w/v) rice polishings after 72 h of incubation at 28°C at pH 4. Carbon and nitrogen ratio of 20 : 1 gave significantly higher production of fungal biomass protein. The FBP in the 75 L fermenter contained 49.50% crude protein, 32.00% true protein, 19.45% crude fiber, 9.62% ash, 11.5% cellulose content, and 0.325% RNA content. The profile of amino acids of final FBP exhibited that all essential amino acids were present in great quantities. The FBP produced by this fungus has been shown to be of good nutritional value for supplementation to poultry. The results presented in this study have practical implications in that the fungus T. harzianum could be used successfully to produce fungal biomass protein using rice polishings.

Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production.

PubMed

Rahnama, Nooshin; Foo, Hooi Ling; Abdul Rahman, Nor Aini; Ariff, Arbakariya; Md Shah, Umi Kalsom

2014-12-12

Rice straw has shown to be a promising agricultural by-product in the bioconversion of biomass to value-added products. Hydrolysis of cellulose, a main constituent of lignocellulosic biomass, is a requirement for fermentable sugar production and its subsequent bioconversion to biofuels such as biobutanol. The high cost of commercial enzymes is a major impediment to the industrial application of cellulases. Therefore, the use of local microbial enzymes has been suggested. Trichoderma harzianum strains are potential CMCase and β-glucosidase producers. However, few researches have been reported on cellulase production by T. harzianum and the subsequent use of the crude cellulase for cellulose enzymatic hydrolysis. For cellulose hydrolysis to be efficiently performed, the presence of the whole set of cellulase components including exoglucanase, endoglucanase, and β-glucosidase at a considerable concentration is required. Biomass recalcitrance is also a bottleneck in the bioconversion of agricultural residues to value-added products. An effective pretreatment could be of central significance in the bioconversion of biomass to biofuels. Rice straw pretreated using various concentrations of NaOH was subjected to enzymatic hydrolysis. The saccharification of rice straw pretreated with 2% (w/v) NaOH using crude cellulase from local T. harzianum SNRS3 resulted in the production of 29.87 g/L reducing sugar and a yield of 0.6 g/g substrate. The use of rice straw hydrolysate as carbon source for biobutanol fermentation by Clostridium acetobutylicum ATCC 824 resulted in an ABE yield, ABE productivity, and biobutanol yield of 0.27 g/g glucose, 0.04 g/L/h and 0.16 g/g glucose, respectively. As a potential β-glucosidase producer, T. harzianum SNRS3 used in this study was able to produce β-glucosidase at the activity of 173.71 U/g substrate. However, for cellulose hydrolysis to be efficient, Filter Paper Activity at a considerable concentration is also required to initiate the

Biodegradation of carbamazepine and clarithromycin by Trichoderma harzianum and Pleurotus ostreatus investigated by liquid chromatography - high-resolution tandem mass spectrometry (FTICR MS-IRMPD).

PubMed

Buchicchio, Alessandro; Bianco, Giuliana; Sofo, Adriano; Masi, Salvatore; Caniani, Donatella

2016-07-01

In this study, the capability of pharmaceutical biodegradation of fungus Trichoderma harzianum was evaluated through the comparison with the well-known biodegradation capability of white-rot fungus Pleurotus ostreatus. The study was performed in aqueous phase under aerobic conditions, using two of the most frequently detected drugs in water bodies: carbamazepine and clarithromycin, with concentrations commonly found in treated wastewater (4μg/l and 0.03μg/l respectively). For the first time, we demonstrated that T. harzianum is able to remove carbamazepine and clarithromycin. The analyses were performed by reversed-phase liquid chromatography/mass spectrometry, using high-resolution Fourier-transform ion cyclotron resonance mass spectrometry upon electrospray ionization in positive ion mode. The high selectivity and mass accuracy provided by high-resolution mass spectrometry, allowed us to identify some unknown metabolites. On the basis of our study, the major metabolites detected in liquid culture treated by T. harzianum were: 14-hydroxy-descladinosyl- and descladinosyl-clarithromycin, which are pharmacologically inactive products not dangerous for the environment. Copyright © 2016 Elsevier B.V. All rights reserved.

A novel method to prepare concentrated conidial biomass formulation of Trichoderma harzianum for seed application.

PubMed

Singh, P C; Nautiyal, C S

2012-12-01

To prepare concentrated formulation of Trichoderma harzianum MTCC-3841 (NBRI-1055) with high colony forming units (CFU), long shelf life and efficient in root colonization by a simple scrapping method. NBRI-1055 spores scrapped from potato dextrose agar plates were used to prepare a concentrated formulation after optimizing carrier material, moisture content and spore harvest time. The process provides an advantage of maintaining optimum moisture level by the addition of water rather than dehydration. The formulation had an initial 11-12 log(10) CFU g(-1). Its concentrated form reduces its application amount by 100 times (10 g 100 kg(-1) seed) and provides 3-4 log(10) CFU seed(-1). Shelf life of the product was experimentally determined at 30 and 40 °C and predicted at other temperatures following Arrhenius equation. The concentrated formulation as compared to similar products provides an extra advantage of smaller packaging for storage and transportation, cutting down product cost. Seed application of the formulation recorded significant increase in plant growth promotion. Stable and effective formulation of Trichoderma harzianum NBRI-1055 was obtained by a simple scrapping method. A new method for the production of concentrated, stable, effective and cost efficient formulation of T. harzianum has been validated for seed application. © 2012 The Society for Applied Microbiology.

Multi-cropping edible truffles and sweet chestnuts: production of high-quality Castanea sativa seedlings inoculated with Tuber aestivum, its ecotype T. uncinatum, T. brumale, and T. macrosporum.

PubMed

Álvarez-Lafuente, Amaya; Benito-Matías, Luis F; Peñuelas-Rubira, Juan L; Suz, Laura M

2018-01-01

The plantation and management of sweet chestnut (Castanea sativa Mill.) orchards is a common and traditional land use system in many areas of Europe that offers the advantage of simultaneous production of nuts and timber. During the last decades, sweet chestnut has declined dramatically in many regions because of the profound social changes in rural areas coupled with pathogen attacks. Truffles, the hypogeous ascocarps of the ectomycorrhizal genus Tuber, are currently cultivated using host trees inoculated with these fungi for improving production in truffle orchards. The production of good forestry quality chestnut seedlings inoculated with European truffles in nurseries is essential for multi-cropping plantation establishment, but so far, it has not been implemented in agroforestry practices. Moreover, it is necessary to assess the physiological condition of the seedlings due to the high calcium amendment needed for the growth of Tuber spp. mycelium that can become toxic for the host plants. In this study, seedlings of C. sativa were inoculated with Tuber aestivum and its ecotypes T. uncinatum, T. brumale, and T. macrosporum and were grown in a greenhouse using culture conditions favorable for the production of high-quality plants for forestry purposes. At the end of the assay, levels of root colonization and morphological and physiological parameters of the seedlings were measured. The colonization of C. sativa with T. aestivum, its ecotype T. uncinatum, and T. brumale was successful, and the seedlings showed normal growth. Inoculation protocols with T. macrosporum need to be improved. Tuber species formed well-developed ectomycorrhizae on C. sativa in nursery conditions.

Transgenic expression of the Trichoderma harzianum hsp70 gene increases Arabidopsis resistance to heat and other abiotic stresses.

PubMed

Montero-Barrientos, Marta; Hermosa, Rosa; Cardoza, Rosa E; Gutiérrez, Santiago; Nicolás, Carlos; Monte, Enrique

2010-05-15

The ability of some Trichoderma strains, a biological control agent, to overcome extreme environmental conditions has previously been reported and related to heat-shock proteins (HSPs). These proteins are induced environmentally and are involved in important processes, acting as molecular chaperones in all organisms. In a previous study, we demonstrated, by overexpression, that the Trichoderma harzianum hsp70 gene conferred tolerance to heat and other abiotic stresses to this fungus. In this work, we investigate the function of the T. harzianum T34 hsp70 gene in Arabidopsis thaliana. We analyze transgenic plant responses under adverse environmental conditions and the expression levels of a set of seven stress genes, using quantitative RT-PCR. As expected, transgenic plants expressing the T. harzianum hsp70 gene exhibited enhanced tolerance to heat stress. In addition, they did not show growth inhibition and, after heat pre-treatment, transgenic seedlings were more tolerant to osmotic, salt and oxidative stresses with respect to the wild-type behavior. Transgenic lines also had increased transcript levels of the Na(+)/H(+) exchanger 1 (SOS1) and ascorbate peroxidase 1 (APX1) genes, involved in salt and oxidative stress responses, respectively. However, the heat-shock factor (HSF) and four HSP genes tested were down-regulated in 35S:hsp70 plants. Overall, our results indicate that hsp70 confers tolerance to heat and other abiotic stresses and that the fungal HSP70 protein acts as a negative regulator of the HSF transcriptional activity in Arabidopsis. (c) 2009 Elsevier GmbH. All rights reserved.

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

USDA-ARS?s Scientific Manuscript database

Trichothecenes are phytotoxic sesquiterpenoid compounds of fungal origin which can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in the biosynthesis of HA is the conversion of farnesyl diphosphate t...

Sequence analysis and gene expression of putative oil palm chitinase and chitinase-like proteins in response to colonization of Ganoderma boninense and Trichoderma harzianum.

PubMed

Yeoh, K-A; Othman, A; Meon, S; Abdullah, F; Ho, C-L

2013-01-01

Chitinases are glycosyl hydrolases that cleave the β-1,4-glycosidic linkages between N-acetylglucosamine residues in chitin which is a major component of fungal cell wall. Plant chitinases hydrolyze fungal chitin to chitin oligosaccharides that serve as elicitors of plant defense system against fungal pathogens. However, plants synthesize many chitinase isozymes and some of them are not pathogenesis-related. In this study, three full-length cDNA sequences encoding a putative chitinase (EgChit3-1) and two chitinase-like proteins (EgChit1-1 and EgChit5-1) have been cloned from oil palm (Elaeis guineensis) by polymerase chain reaction (PCR). The abundance of these transcripts in the roots and leaves of oil palm seedlings treated with Ganoderma boninense (a fungal pathogen) or Trichoderma harzianum (an avirulent symbiont), and a combination of both fungi at 3, 6 and 12 weeks post infection were profiled by real time quantitative reverse-transcription (qRT)-PCR. Our findings showed that the gene expression of EgChit3-1 increased significantly in the roots of oil palm seedlings treated with either G. boninense or T. harzianum and a combination of both; whereas the gene expression of EgChit1-1 in the treated roots of oil palm seedlings was not significantly higher compared to those of the untreated oil palm roots. The gene expression of EgChit5-1 was only higher in the roots of oil palm seedlings treated with T. harzianum compared to those of the untreated oil palm roots. In addition, the gene expression of EgChit1-1 and EgChit3-1 showed a significantly higher gene expression in the leaf samples of oil palm seedlings treated with either G. boninense or T. harzianum.

Physiological and biochemical characterization of Trichoderma harzianum, a biological control agent against soilborne fungal plant pathogens.

PubMed Central

Grondona, I; Hermosa, R; Tejada, M; Gomis, M D; Mateos, P F; Bridge, P D; Monte, E; Garcia-Acha, I

1997-01-01

Monoconidial cultures of 15 isolates of Trichoderma harzianum were characterized on the basis of 82 morphological, physiological, and biochemical features and 99 isoenzyme bands from seven enzyme systems. The results were subjected to numerical analysis which revealed four distinct groups. Representative sequences of the internal transcribed spacer 1 (ITS 1)-ITS 2 region in the ribosomal DNA gene cluster were compared between groups confirming this distribution. The utility of the groupings generated from the morphological, physiological, and biochemical data was assessed by including an additional environmental isolate in the electrophoretic analysis. The in vitro antibiotic activity of the T. harzianum isolates was assayed against 10 isolates of five different soilborne fungal plant pathogens: Aphanomyces cochlioides, Rhizoctonia solani, Phoma betae, Acremonium cucurbitacearum, and Fusarium oxysporum f. sp. radicis lycopersici. Similarities between levels and specificities of biological activity and the numerical characterization groupings are both discussed in relation to antagonist-specific populations in known and potential biocontrol species. PMID:9251205

Identification of differentially expressed genes in response to mercury I and II stress in Trichoderma harzianum.

PubMed

Puglisi, Ivana; Faedda, Roberto; Sanzaro, Vincenzo; Lo Piero, Angela R; Petrone, Goffredo; Cacciola, Santa O

2012-09-15

Filamentous fungi are very promising organisms in both the control and the reduction of the amount of heavy metal released by human and industrial activities. In particular, Trichoderma harzianum demonstrated to be tolerant towards different heavy metals, such as mercury and cadmium, even though the mechanism underlying this tolerance is not fully understood. By using a particular strategy of the suppression subtractive hybridization technique, we were able to identify in the strain IMI 393899 of T. harzianum eight different genes up-regulated in the presence of mercury II with respect to cadmium. Among the genes identified, a possible role in the tolerance mechanism could be envisaged for hydrophobin, due to its ability to dissolve hydrophobic molecules into aqueous media. We also show that IMI 393899 grows at the same rate of control culture in the presence of mercury I and that all eight genes isolated were also up-regulated in this condition. Copyright © 2012 Elsevier B.V. All rights reserved.

Gene expression analysis of the biocontrol fungus Trichoderma harzianum in the presence of tomato plants, chitin, or glucose using a high-density oligonucleotide microarray.

PubMed

Samolski, Ilanit; de Luis, Alberto; Vizcaíno, Juan Antonio; Monte, Enrique; Suárez, M Belén

2009-10-13

It has recently been shown that the Trichoderma fungal species used for biocontrol of plant diseases are capable of interacting with plant roots directly, behaving as symbiotic microorganisms. With a view to providing further information at transcriptomic level about the early response of Trichoderma to a host plant, we developed a high-density oligonucleotide (HDO) microarray encompassing 14,081 Expressed Sequence Tag (EST)-based transcripts from eight Trichoderma spp. and 9,121 genome-derived transcripts of T. reesei, and we have used this microarray to examine the gene expression of T. harzianum either alone or in the presence of tomato plants, chitin, or glucose. Global microarray analysis revealed 1,617 probe sets showing differential expression in T. harzianum mycelia under at least one of the culture conditions tested as compared with one another. Hierarchical clustering and heat map representation showed that the expression patterns obtained in glucose medium clustered separately from the expression patterns observed in the presence of tomato plants and chitin. Annotations using the Blast2GO suite identified 85 of the 257 transcripts whose probe sets afforded up-regulated expression in response to tomato plants. Some of these transcripts were predicted to encode proteins related to Trichoderma-host (fungus or plant) associations, such as Sm1/Elp1 protein, proteases P6281 and PRA1, enchochitinase CHIT42, or QID74 protein, although previously uncharacterized genes were also identified, including those responsible for the possible biosynthesis of nitric oxide, xenobiotic detoxification, mycelium development, or those related to the formation of infection structures in plant tissues. The effectiveness of the Trichoderma HDO microarray to detect different gene responses under different growth conditions in the fungus T. harzianum strongly indicates that this tool should be useful for further assays that include different stages of plant colonization, as well as

Gene expression analysis of the biocontrol fungus Trichoderma harzianum in the presence of tomato plants, chitin, or glucose using a high-density oligonucleotide microarray

PubMed Central

2009-01-01

Background It has recently been shown that the Trichoderma fungal species used for biocontrol of plant diseases are capable of interacting with plant roots directly, behaving as symbiotic microorganisms. With a view to providing further information at transcriptomic level about the early response of Trichoderma to a host plant, we developed a high-density oligonucleotide (HDO) microarray encompassing 14,081 Expressed Sequence Tag (EST)-based transcripts from eight Trichoderma spp. and 9,121 genome-derived transcripts of T. reesei, and we have used this microarray to examine the gene expression of T. harzianum either alone or in the presence of tomato plants, chitin, or glucose. Results Global microarray analysis revealed 1,617 probe sets showing differential expression in T. harzianum mycelia under at least one of the culture conditions tested as compared with one another. Hierarchical clustering and heat map representation showed that the expression patterns obtained in glucose medium clustered separately from the expression patterns observed in the presence of tomato plants and chitin. Annotations using the Blast2GO suite identified 85 of the 257 transcripts whose probe sets afforded up-regulated expression in response to tomato plants. Some of these transcripts were predicted to encode proteins related to Trichoderma-host (fungus or plant) associations, such as Sm1/Elp1 protein, proteases P6281 and PRA1, enchochitinase CHIT42, or QID74 protein, although previously uncharacterized genes were also identified, including those responsible for the possible biosynthesis of nitric oxide, xenobiotic detoxification, mycelium development, or those related to the formation of infection structures in plant tissues. Conclusion The effectiveness of the Trichoderma HDO microarray to detect different gene responses under different growth conditions in the fungus T. harzianum strongly indicates that this tool should be useful for further assays that include different stages of

Biodegradation of Mordant orange-1 using newly isolated strain Trichoderma harzianum RY44 and its metabolite appraisal.

PubMed

Hadibarata, Tony; Syafiuddin, Achmad; Al-Dhabaan, Fahad A; Elshikh, Mohamed Soliman; Rubiyatno

2018-05-01

Herein, we systematically reported the capability of T. harzianum RY44 for decolorization of Mordant orange-1. The fungi strains were isolated from the Universiti Teknologi Malaysia tropical rain forest. For initial screening, the decolorization was conducted using 50 strains of the fungi for 20 days incubation time and the best performance was selected. Then, the decolorization capability and fungal biomass were evaluated using different dye concentrations, namely, 0, 50, 75 and 100 ppm. Effects of the carbon sources (fructose, glucose, and galactose), nitrogen sources (ammonium nitrate, ammonium sulfate and yeast extract), surfactant (tween 80), aromatic compounds (benzoic acid, catechol and salicylic acid), and pH on the decolorization efficiency were examined. This study has found that the employed carbon sources, nitrogen sources, and aromatic compounds strongly enhance the decolorization efficiency. In addition, increasing the surfactant volume and pH generally decreased the decolorization efficiencies from 19.5 to 9.0% and 81.7 to 60.5%, respectively. In the mechanism philosophy, the present work has found that Mordant orange-1 were initially degraded by T. harzianum RY44 to benzoic acid and finally transformed into salicylic acid.

Contribution of the saprobic fungi Trametes versicolor and Trichoderma harzianum and the arbuscular mycorrhizal fungi Glomus deserticola and G. claroideum to arsenic tolerance of Eucalyptus globulus.

PubMed

Arriagada, C; Aranda, E; Sampedro, I; Garcia-Romera, I; Ocampo, J A

2009-12-01

The presence of high concentrations of arsenic (As) decreased the shoot and root dry weight, chlorophyll and P and Mg content of Eucalyptus globulus colonized with the arbuscular mycorrhizal (AM) fungi Glomus deserticola or G. claroideum, but these parameters were higher than in non-AM plants. As increased the percentage of AM length colonization and succinate dehydrogenase (SDH) activity in the root of E. globulus. Trichoderma harzianum, but not Trametes versicolor, increased the shoot and root dry weight, chlorophyll content, the percentage of AM root length colonization and SDH activity of E. globulus in presence of all As concentrations applied to soil when was inoculated together with G. claroideum. AM fungi increased shoot As and P concentration of E. globulus to higher level than the non-AM inoculated controls. The contribution of the AM and saprobe fungi to the translocation of As from root to shoot of E. globulus is discussed.

Bio-protective microbial agents from rhizosphere eco-systems trigger plant defense responses provide protection against sheath blight disease in rice (Oryza sativa L.).

PubMed

Singh, Udai B; Malviya, Deepti; Wasiullah; Singh, Shailendra; Pradhan, Jatindra K; Singh, Bhanu P; Roy, Manish; Imram, Mohd; Pathak, Neelam; Baisyal, B M; Rai, Jai P; Sarma, B K; Singh, Rajiv K; Sharma, P K; Kaur, Saman Deep; Manna, M C; Sharma, Sushil K; Sharma, Arun K

2016-11-01

Sheath blight of rice (Oryza sativa L.) caused by Rhizoctonia solani is a major disease and attempts are being made to develop microbe based technologies for biocontrol of this pathogen. However, the mechanisms of biocontrol are not fully understood and still require indepth study in the backdrop of emerging concepts in biological systems. The present investigation was aimed at deciphering the mechanisms of biocontrol of sheath blight of rice employing Pseudomonas fluorescens and Trichoderma harzianum as model agents for biocontrol. Initially 25, 5 and 5 strains of P. fluorescens, T. viride and T. harzianum, respectively, were screened for their biocontrol potential. Out of which, six strains with higher value of percent inhibition of fungal mycelium in dual plate assay were selected. The role of P. fluorescens, T. viride and T. harzianum were investigated in induction and bioaccumulation of natural antioxidants, defence-related biomolecules and other changes in plant which lead not only to growth promotion but also protection from pathogenic stress conditions in rice. The two most promising strains, P. fluorescens PF-08 and T. harzianum UBSTH-501 selected on the basis of in planta evaluation, when applied individually or in combination, significantly enhanced the accumulation of defence-related biomolecules, enzymes and exhibited biocontrol potential against R. solani. A modified/newly developed delivery system was applied for the first time in the experiments involving inoculation of plants with both bioagents, viz. P. fluorescens PF-08 and T. harzianum UBSTH-501. Results suggested that application of P. fluorescens PF-08 and T. harzianum UBSTH-501 alone or in combination, not only helps in control of the disease but also increases plant growth along with reduction in application of toxic chemical pesticides. Copyright © 2016 Elsevier GmbH. All rights reserved.

Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

PubMed

Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

2016-03-01

Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

Evaluation of Trichoderma spp. for biocontrol of tomato sudden caused by Pythium aphanidermatum following flooding in tropical hot season.

PubMed

Le, H T T; Black, L L; Sikora, R A

2003-01-01

Tomato sudden death is a major problem in tomato production in tropical lowland areas. The plant wilts and dies following artificial or natural flooding for 48-72 hrs in the summer season. Occurrence of this disease is related to aggressiveness of Pythium aphanidermatum on tomato at high soil temperature (>30 degrees C). Several methods such as using biological control agents, fungicides and other cultural practices were applied in attempts to control tomato sudden death. Three Trichoderma harzianum and two Trichoderma virens isolates were evaluated for biocontrol of the disease in the greenhouse and in the field T. harzianum and T. virens isolates were separately used to treat the seed, potting medium and also incorporated into the soil before transplanting. Field soil was naturally infested with P. aphanidermatum, while greenhouse soil was inoculated with the pathogen 10 days after transplanting. All treatments were flooded for 48 hrs at 32 degrees C soil temperature. Results from this study show that most tested T. harzianum and T. virens isolates have little promise for control of tomato sudden death following flooding. The percentage of tomato plants that wilted after growing in soil treated with either T. harzianum or T. virens and P. aphanidermatum was not significantly different when compared to the soil treated with P. aphanidermatum alone. No wilted plants were observed in the control (non treated soil).

Recombinant Trichoderma harzianum endoglucanase I (Cel7B) is a highly acidic and promiscuous carbohydrate-active enzyme.

PubMed

Pellegrini, Vanessa O A; Serpa, Viviane Isabel; Godoy, Andre S; Camilo, Cesar M; Bernardes, Amanda; Rezende, Camila A; Junior, Nei Pereira; Franco Cairo, João Paulo L; Squina, Fabio M; Polikarpov, Igor

2015-11-01

Trichoderma filamentous fungi have been investigated due to their ability to secrete cellulases which find various biotechnological applications such as biomass hydrolysis and cellulosic ethanol production. Previous studies demonstrated that Trichoderma harzianum IOC-3844 has a high degree of cellulolytic activity and potential for biomass hydrolysis. However, enzymatic, biochemical, and structural studies of cellulases from T. harzianum are scarce. This work reports biochemical characterization of the recombinant endoglucanase I from T. harzianum, ThCel7B, and its catalytic core domain. The constructs display optimum activity at 55 °C and a surprisingly acidic pH optimum of 3.0. The full-length enzyme is able to hydrolyze a variety of substrates, with high specific activity: 75 U/mg for β-glucan, 46 U/mg toward xyloglucan, 39 U/mg for lichenan, 26 U/mg for carboxymethyl cellulose, 18 U/mg for 4-nitrophenyl β-D-cellobioside, 16 U/mg for rye arabinoxylan, and 12 U/mg toward xylan. The enzyme also hydrolyzed filter paper, phosphoric acid swollen cellulose, Sigmacell 20, Avicel PH-101, and cellulose, albeit with lower efficiency. The ThCel7B catalytic domain displays similar substrate diversity. Fluorescence-based thermal shift assays showed that thermal stability is highest at pH 5.0. We determined kinetic parameters and analyzed a pattern of oligosaccharide substrates hydrolysis, revealing cellobiose as a final product of C6 degradation. Finally, we visualized effects of ThCel7B on oat spelt using scanning electron microscopy, demonstrating the morphological changes of the substrate during the hydrolysis. The acidic behavior of ThCel7B and its considerable thermostability hold a promise of its industrial applications and other biotechnological uses under extremely acidic conditions.

Integrated effect of microbial antagonist, organic amendment and fungicide in controlling seedling mortality (Rhizoctonia solani) and improving yield in pea (Pisum sativum L.).

PubMed

Akhter, Wasira; Bhuiyan, Mohamed Khurshed Alam; Sultana, Farjana; Hossain, Mohamed Motaher

2015-01-01

The study evaluated the comparative performance of a few microbial antagonists, organic amendments and fungicides and their integration for the management of seedling mortality (Rhizoctonia solani Kühn) and yield improvement in pea (Pisum sativum L.). Before setting the experiment in field microplots, a series of in vitro and in vivo experiments were conducted to select a virulent isolate of R. solani, an effective antagonistic isolate of Trichoderma harzianum, a fungitoxic organic amendment and an appropriate fungicide. A greenhouse pathogenicity test compared differences in seedling mortality in pea inoculated by four isolates of R. solani and identified the isolate RS10 as the most virulent one. Among the 20 isolates screened in dual culture assay on PDA, T. harzianum isolate T-3 was found to show the highest (77.22%) inhibition of the radial growth of R. solani. A complete inhibition (100.00%) of colony growth of R. solani was observed when fungicide Bavistin 50 WP and Provax-200 at the rate of 100 and 250 ppm, respectively, were used, while Provax-200 was found to be highly compatible with T. harzianum. Mustard oilcake gave maximum inhibition (60.28%) of the radial growth of R. solani at all ratios, followed by sesame oilcake and tea waste. Integration of soil treatment with T. harzianum isolate T-3 and mustard oilcake and seed treatment with Provax-200 appeared to be significantly superior in reducing seedling mortality and improving seed yield in pea in comparison to any single or dual application of them in the experimental field. The research results will help growers develop integrated disease management strategies for the control of Rhizoctonia disease in pea. The research results show the need for an integrating selective microbial antagonist, organic amendment and fungicide to achieve appropriate management of seedling mortality (R. solani) and increase of seed yield in pea. Copyright © 2014 Académie des sciences. Published by Elsevier SAS. All

Effectiveness of T. harzianum and Humate Amendment in Soil Salinity Restoration

NASA Astrophysics Data System (ADS)

Apostolakis, Antonios; Daliakopoulos, Ioannis; Tsanis, Ioannis

2017-04-01

Soil salinity is a major soil degradation threat, especially for the water stressed parts of the Mediterranean region, where it hinders soil fertility and thus agricultural productivity. Soil salinity management can be complex and expensive, often resorting to the use of chemical amendments thus risking soil and aquifer pollution. This study quantifies the beneficial effects of (a) a commercial strain of the beneficial fungus Trichoderma harzianum (TH), and (b) a commercial humate fertilizer enhancer (HFE) approved for organic farming, against soil salinization. The treatments are tested in the context of a Solanum lycopersicum (tomato) greenhouse simulation of the cultivation conditions typical for the semi-arid coastal Timpaki basin in south-central Crete. 20 vigorous 20-day-old Solanum lycopersicum L. cv Elpida seedlings are treated either with TH or HFE, using soil substrates and irrigation treatments of two degradation states. 20 additional plants serve either as controls or guard rows. All plants are transplanted into 35 L pots under greenhouse conditions. Preliminary analysis of soil salinity and crop yield indicators suggest that both treatments are beneficial for the soil-plant system, each to a different extent depending on initial soil conditions.

Biological Role of Trichoderma harzianum-Derived Platelet-Activating Factor Acetylhydrolase (PAF-AH) on Stress Response and Antagonism

PubMed Central

Yu, Chuanjin; Fan, Lili; Wu, Qiong; Fu, Kehe; Gao, Shigang; Wang, Meng; Gao, Jinxin; Li, Yaqian; Chen, Jie

2014-01-01

We investigated the properties of platelet-activating factor acetylhydrolase (PAF-AH) derived from Trichoderma harzianum. The enzyme, comprised of 572 amino acids, shares high homology with PAF-AH proteins from T. koningii and other microbial species. The optimum enzymatic activity of PAF-AH occurred at pH 6 in the absence of Ca2+ and it localized in the cytoplasm, and we observed the upregulation of PAF-AH expression in response to carbon starvation and strong heat shock. Furthermore, PAF-AH knockout transformant growth occurred more slowly than wild type cells and over-expression strains grown in SM medium at 37°C and 42°C. In addition, PAF-AH expression significantly increased under a series of maize root induction assay. Eicosanoic acid and ergosterol levels decreased in the PAF-AH knockouts compared to wild type cells, as revealed by GC/MS analysis. We also determined stress responses mediated by PAF-AH were related to proteins HEX1, Cu/Zn superoxide dismutase, and cytochrome c. Finally, PAF-AH exhibited antagonistic activity against Rhizoctonia solani in plate confrontation assays. Our results indicate PAF-AH may play an important role in T. harzianum stress response and antagonism under diverse environmental conditions. PMID:24964161

Biological role of Trichoderma harzianum-derived platelet-activating factor acetylhydrolase (PAF-AH) on stress response and antagonism.

PubMed

Yu, Chuanjin; Fan, Lili; Wu, Qiong; Fu, Kehe; Gao, Shigang; Wang, Meng; Gao, Jinxin; Li, Yaqian; Chen, Jie

2014-01-01

We investigated the properties of platelet-activating factor acetylhydrolase (PAF-AH) derived from Trichoderma harzianum. The enzyme, comprised of 572 amino acids, shares high homology with PAF-AH proteins from T. koningii and other microbial species. The optimum enzymatic activity of PAF-AH occurred at pH 6 in the absence of Ca2+ and it localized in the cytoplasm, and we observed the upregulation of PAF-AH expression in response to carbon starvation and strong heat shock. Furthermore, PAF-AH knockout transformant growth occurred more slowly than wild type cells and over-expression strains grown in SM medium at 37°C and 42°C. In addition, PAF-AH expression significantly increased under a series of maize root induction assay. Eicosanoic acid and ergosterol levels decreased in the PAF-AH knockouts compared to wild type cells, as revealed by GC/MS analysis. We also determined stress responses mediated by PAF-AH were related to proteins HEX1, Cu/Zn superoxide dismutase, and cytochrome c. Finally, PAF-AH exhibited antagonistic activity against Rhizoctonia solani in plate confrontation assays. Our results indicate PAF-AH may play an important role in T. harzianum stress response and antagonism under diverse environmental conditions.

Functional Analysis of the Trichoderma harzianum nox1 Gene, Encoding an NADPH Oxidase, Relates Production of Reactive Oxygen Species to Specific Biocontrol Activity against Pythium ultimum▿†

PubMed Central

Montero-Barrientos, M.; Hermosa, R.; Cardoza, R. E.; Gutiérrez, S.; Monte, E.

2011-01-01

The synthesis of reactive oxygen species (ROS) is one of the first events following pathogenic interactions in eukaryotic cells, and NADPH oxidases are involved in the formation of such ROS. The nox1 gene of Trichoderma harzianum was cloned, and its role in antagonism against phytopathogens was analyzed in nox1-overexpressed transformants. The increased levels of nox1 expression in these transformants were accompanied by an increase in ROS production during their direct confrontation with Pythium ultimum. The transformants displayed an increased hydrolytic pattern, as determined by comparing protease, cellulase, and chitinase activities with those for the wild type. In confrontation assays against P. ultimum the nox1-overexpressed transformants were more effective than the wild type, but not in assays against Botrytis cinerea or Rhizoctonia solani. A transcriptomic analysis using a Trichoderma high-density oligonucleotide (HDO) microarray also showed that, compared to gene expression for the interaction of wild-type T. harzianum and P. ultimum, genes related to protease, cellulase, and chitinase activities were differentially upregulated in the interaction of a nox1-overexpressed transformant with this pathogen. Our results show that nox1 is involved in T. harzianum ROS production and antagonism against P. ultimum. PMID:21421791

Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme.

PubMed

Nascimento, Alessandro S; Muniz, Joao Renato C; Aparício, Ricardo; Golubev, Alexander M; Polikarpov, Igor

2014-09-01

Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. β-Mannosidases are distributed between glycoside hydrolase (GH) families 1, 2, and 5, and only a handful of the enzymes have been structurally characterized to date. The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. No structures of eukaryotic mannosidases of this family are currently available. To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. © 2014 FEBS.

Wetting properties of fungi mycelium alter soil infiltration and soil water repellency in a γ-sterilized wettable and repellent soil.

PubMed

Chau, Henry Wai; Goh, Yit Kheng; Vujanovic, Vladimir; Si, Bing Cheng

2012-12-01

Soil water repellency (SWR) has a drastic impact on soil quality resulting in reduced infiltration, increased runoff, increased leaching, reduced plant growth, and increased soil erosion. One of the causes of SWR is hydrophobic fungal structures and exudates that change the soil-water relationship. The objective of this study was to determine whether SWR and infiltration could be manipulated through inoculation with fungi. The effect of fungi on SWR was investigated through inoculation of three fungal strains (hydrophilic -Fusarium proliferatum, chrono-amphiphilic -Trichoderma harzianum, and hydrophobic -Alternaria sp.) on a water repellent soil (WR-soil) and a wettable soil (W-soil). The change in SWR and infiltration was assessed by the water repellency index and cumulative infiltration respectively. F. proliferatum decreased the SWR on WR-soil and slightly increased SWR in W-soil, while Alternaria sp. increased SWR in both the W-soil and the WR-soil. Conversely T. harzianum increased the SWR in the W-soil and decreased the SWR in the WR-soil. All strains showed a decrease in infiltration in W-soil, while only the F. proliferatum and T. harzianum strain showed improvement in infiltration in the WR-soil. The ability of fungi to alter the SWR and enmesh soil particles results in changes to the infiltration dynamics in soil. Copyright © 2012 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Structure and properties of native and unfolded lysing enzyme from T. harzianum: Chemical and pH denaturation.

PubMed

Bey, Houda; Gtari, Wala; Aschi, Adel; Othman, Tahar

2016-11-01

The effect of chemical denaturants and pH on the change of the conformation of the protein Lysing Enzyme from Trichoderma Harzianum has been investigated by dynamic light scattering (DLS) and turbidimetry. Chemical denaturants are frequently used to describe the mechanisms of folding and transition states. We have analyzed the pH effect on the properties and particle size of the protein. The compaction factor CI has shown that the protein is weakly disordered. The molecular dynamics simulations confirm, at neutral pH, that the protein has a low net charge and high hydrophobicity. Copyright © 2016 Elsevier B.V. All rights reserved.

Experimental mixture design as a tool to enhance glycosyl hydrolases production by a new Trichoderma harzianum P49P11 strain cultivated under controlled bioreactor submerged fermentation.

PubMed

Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; Lima, Deise Juliana da Silva; Pradella, José Geraldo da Cruz

2013-03-01

This work investigates the glycosyl hydrolase (GH) profile of a new Trichoderma harzianum strain cultivated under controlled bioreactor submerged fermentation. The influence of different medium components (delignified steam-exploded sugarcane bagasse, sucrose, and soybean flour) on GH biosynthesis was assessed using experimental mixture design (EMD). Additionally, the effect of increased component concentrations in culture media selected from the EMD was studied. It was found that that a mixed culture medium could significantly maximize GH biosynthesis rate, especially for xylanase enzymes which achieved a 2-fold increment. Overall, it was demonstrated that T. harzianumP49P11 enzymes have a great potential to be used in the deconstruction of biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens.

PubMed

Vinale, F; Marra, R; Scala, F; Ghisalberti, E L; Lorito, M; Sivasithamparam, K

2006-08-01

Trichoderma harzianum strains T22 and T39 are two micro-organisms used as active agents in a variety of commercial biopesticides and biofertilizers and widely applied amongst field and greenhouse crops. The production, isolation, biological and chemical characterization of the main secondary metabolites produced by these strains are investigated. Of the three major compounds produced by strain T22, one is a new azaphilone that shows marked in vitro inhibition of Rhizoctonia solani, Pythium ultimum and Gaeumannomyces graminis var. tritici. In turn, filtrates from strain T39 were demonstrated to contain two compounds previously isolated from other T. harzianum strains and a new butenolide. The production of the isolated metabolites was also monitored by liquid chromatography/mass spectrometry during in vitro interaction with R. solani. This paper reports the isolation and characterization of the main secondary metabolites obtained from culture filtrates of two T. harzianum strains and their production during antagonistic interaction with the pathogen R. solani. This is the first work on secondary metabolites produced by the commercially applied strains T22 and T39. Our results provide a better understanding of the metabolism of these fungi, which are both widely used as biopesticides and/or biofertilizers in biocontrol.

Characterization of a chitinase and an endo-beta-1,3-glucanase from Trichoderma harzianum Rifai T24 involved in control of the phytopathogen Sclerotium rolfsii.

PubMed

El-Katatny, M H; Gudelj, M; Robra, K H; Elnaghy, M A; Gübitz, G M

2001-07-01

Of 24 Trichoderma isolates, T harzianum Rifai (T24) showed a potential for control of the phytopathogenic basidiomycete Sclerotium rolfsii. When T24 was grown on different carbon sources, growth inhibition of S. rolfsii by the T24 culture filtrate correlated with the activity of extracellular chitinase and beta-1,3-glucanase. The 43-kilodalton (kDa) chitinase and the 74-kDa beta-1,3-glucanase were purified from the T24 culture filtrate in two and three steps, respectively, using ammonium sulphate precipitation followed by hydrophobic interaction chromatography (phenyl-Sepharose) and gel filtration (beta-1,3-glucanase). Km and Kcat were 3.8 g l(-1) and 0.71 s(-1) for the chitinase (chitin) and 1.1 g(-1) and 52 s(-1) for the beta-1,3-glucanase (laminarin). The chitinase showed higher activity on chitin than on less-acetylated substrate analogues (chitosan), while the beta-1,3-glucanase was specific for beta-1,3-linkages in polysaccharides. Both enzymes were stable at 30 degrees C, while at 60 degrees C the chitinase and the beta-1,3-glucanase were rapidly inactivated, showing half-lives of 15 and 20 min, respectively. The enzymes inhibited growth of S. rolfsii in an additive manner showing a promising ED50 (50% effective dose) value of 2.7 microg/ml.

Enzyme Diffusion from Trichoderma atroviride (= T. harzianum P1) to Rhizoctonia solani Is a Prerequisite for Triggering of Trichoderma ech42 Gene Expression before Mycoparasitic Contact

PubMed Central

Kullnig, Cornelia; Mach, Robert L.; Lorito, Matteo; Kubicek, Christian P.

2000-01-01

A plate confrontation experiment is commonly used to study the mechanism by which Trichoderma spp. antagonize and parasitize other fungi. Previous work with chitinase gene expression (ech42) during the precontact period of this process in which cellophane and dialysis membranes separated Trichoderma harzianum and its host Rhizoctonia solani resulted in essentially opposite results. Here, we show that cellophane membranes are permeable to proteins up to at least 90 kDa in size but that dialysis membranes are not. ech42 was expressed during the precontact stage of the confrontation between Trichoderma atroviride and its host only if the cellophane was placed between the two fungi. These results are consistent with enzyme diffusion from T. atroviride to R. solani generating the trigger of ech42 gene expression. PMID:10788407

Trichoderma-Based Biostimulants Modulate Rhizosphere Microbial Populations and Improve N Uptake Efficiency, Yield, and Nutritional Quality of Leafy Vegetables

PubMed Central

Fiorentino, Nunzio; Ventorino, Valeria; Woo, Sheridan L.; Pepe, Olimpia; De Rosa, Armando; Gioia, Laura; Romano, Ida; Lombardi, Nadia; Napolitano, Mauro; Colla, Giuseppe; Rouphael, Youssef

2018-01-01

Microbial inoculants such as Trichoderma-based products are receiving great interest among researchers and agricultural producers for their potential to improve crop productivity, nutritional quality as well as resistance to plant pathogens/pests and numerous environmental stresses. Two greenhouse experiments were conducted to assess the effects of Trichoderma-based biostimulants under suboptimal, optimal and supraoptimal levels of nitrogen (N) fertilization in two leafy vegetables: Iceberg lettuce (Lactuca sativa L.) and rocket (Eruca sativa Mill.). The yield, nutritional characteristics, N uptake and mineral composition were analyzed for each vegetable crop after inoculation with Trichoderma strains T. virens (GV41) or T. harzianum (T22), and results were compared to non-inoculated plants. In addition, the effect of the Trichoderma-based biostimulants on microbes associated with the rhizosphere in terms of prokaryotic and eukaryotic composition and concentration using DGGE was also evaluated. Trichoderma-based biostimulants, in particular GV41, positively increased lettuce and rocket yield in the unfertilized plots. The highest marketable lettuce fresh yield was recorded with either of the biostimulant inoculations when plants were supplied with optimal levels of N. The inoculation of rocket with GV41, and to a lesser degree with T22, elicited an increase in total ascorbic acid under both optimal and high N conditions. T. virens GV41 increased N-use efficiency of lettuce, and favored the uptake of native N present in the soil of both lettuce and rocket. The positive effect of biostimulants on nutrient uptake and crop growth was species-dependent, being more marked with lettuce. The best biostimulation effects from the Trichoderma treatments were observed in both crops when grown under low N availability. The Trichoderma inoculation strongly influenced the composition of eukaryotic populations in the rhizosphere, in particularly exerting different effects with low

Identification and Characterization of Trichoderma Species Damaging Shiitake Mushroom Bed-Logs Infested by Camptomyia Pest.

PubMed

Kim, Jun Young; Kwon, Hyuk Woo; Yun, Yeo Hong; Kim, Seong Hwan

2016-05-28

The shiitake mushroom industry has suffered from Camptomyia (gall midges) pest, which feeds on the mycelium of shiitake mushroom during its cultivation. It has been postulated that fungal damage of shiitake bed-logs is associated with infestation by the insect pest, but this is not well understood. To understand the fungal damage associated with Camptomyia pest, various Trichoderma species were isolated, identified, and characterized. In addition to two previously known Trichoderma species, T. citrinoviride and T. deliquescens, two other Trichoderma species, T. harzianum and T. atroviride, were newly identified from the pestinfested bed-log samples obtained at three mushroom farms in Cheonan, Korea. Among these four species, T. harzianum was the most evident. The results of a chromogenic media-based assay for extracellular enzymes showed that these four species have the ability to produce amylase, carboxyl-methyl cellulase, avicelase, pectinase, and β-glucosidase, thus indicating that they can degrade wood components. A dual culture assay on PDA indicated that T. harzianum, T. atroviride, and T. citrinoviride were antagonistic against the mycelial growth of a shiitake strain (Lentinula edodes). Inoculation tests on shiitake bed-logs revealed that all four species were able to damage the wood of bed-logs. Our results provide evidence that the four green mold species are the causal agents involved in fungal damage of shiitake bed-logs infested by Camptomyia pest.

Application of Trichoderma harzianum SQR-T037 bio-organic fertiliser significantly controls Fusarium wilt and affects the microbial communities of continuously cropped soil of cucumber.

PubMed

Chen, Li-Hua; Huang, Xin-Qi; Zhang, Feng-Ge; Zhao, Di-Kun; Yang, Xing-Ming; Shen, Qi-Rong

2012-09-01

The reduction in diversity of the soil microbial community causes the disorder of continuous cropping. The aim of this study was to determine the effects of applying Trichoderma harzianum SQR-T037 bio-organic fertiliser (BIO) on the microbial community in continuously cropped cucumber soil. Four treatments were set: (1) control, where neither seedling nursery soil (N) nor transplanted soil (T) was amended with BIO; (2) N treatment, where nursery soil was amended with BIO (1% w/w) but transplanted soil was not; (3) N + T treatment, where BIO was added to both nursery soil (1% w/w) and transplanted soil (0.5% w/w); (4) uncropped soil, where soil was left uncropped consistently. A disease index of 72.2% was found for the control treatment, while the N and N + T treatments had disease indices of only 25 and 15% respectively. Analysis of the denaturing gradient gel electrophoresis (DGGE) profiles showed that the bacterial communities of the N and N + T treatments were similar to those of the uncropped soil but distinct from those of the control soil. The fungal communities of the N and N + T treatments differed from those of both the uncropped soil and the control. Addition of BIO to both the nursery soil and the transplanted soil can diversify the microbial community in continuously cropped cucumber soil and thus effectively control Fusarium wilt of cucumber plants. Copyright © 2012 Society of Chemical Industry.

Re-inoculation strategies enhance the degradation of emerging pollutants in fungal bioaugmentation of sewage sludge.

PubMed

Rodríguez-Rodríguez, Carlos E; Lucas, Daniel; Barón, Enrique; Gago-Ferrero, Pablo; Molins-Delgado, Daniel; Rodríguez-Mozaz, Sara; Eljarrat, Ethel; Díaz-Cruz, M Silvia; Barceló, Damià; Caminal, Glòria; Vicent, Teresa

2014-09-01

The use of Trametes versicolor has been partially successful in the removal of some pharmaceuticals from sewage sludge in laboratory-scale biopile systems. The application of two strategies for the re-inoculation of biomass was assessed during the fungal bioaugmentation of non-sterile sludge (42-d treatment) as an approach to improve the elimination of pharmaceuticals and other groups of emerging pollutants. Globally, the re-inoculation of biopiles with blended mycelium exerted a major effect on the removal of pharmaceuticals (86%), brominated-flame-retardants (81%) and UV filters (80%) with respect to the re-inoculation with additional lignocellulosic substrate colonized by the fungus (69-67-22%). The performance was better than that of the analogous non-re-inoculated systems that were assayed previously for the removal of pharmaceuticals. The results demonstrate the ability of T. versicolor to remove a wide spectrum of emerging micropollutants under non-sterile conditions, while re-inoculation appears to be a useful step to improve the fungal treatment of sludge. Copyright © 2014 Elsevier Ltd. All rights reserved.

Construction and functional analysis of Trichoderma harzianum mutants that modulate maize resistance to the pathogen Curvularia lunata.

PubMed

Fan, Lili; Fu, Kehe; Yu, Chuanjin; Ma, Jia; Li, Yaqian; Chen, Jie

2014-01-01

Agrobacterium tumefaciens-mediated transformation (ATMT) was used to generate an insertional mutant library of the mycelial fungus Trichoderma harzianum. From a total of 450 mutants, six mutants that showed significant influence on maize resistance to C. lunata were analyzed in detail. Maize coated with these mutants was more susceptible to C. lunata compared with those coated with a wild-type (WT) strain. Similar to other fungal ATMT libraries, all six mutants were single copy integrations, which occurred preferentially in noncoding regions (except two mutants) and were frequently accompanied by the loss of border sequences. Two mutants (T66 and T312) that were linked to resistance were characterized further. Maize seeds coated with T66 and T312 were more susceptible to C. lunata than those treated with WT. Moreover, the mutants affected the resistance of maize to C. lunata by enhancing jasmonate-responsive gene expression. T66 and T312 induced maize resistance to C. lunata infection through a jasmonic acid-dependent pathway.

Production of Lytic Enzymes by Trichoderma Isolates during in vitro Antagonism with Aspergillus Niger, The Causal Agent of Collar ROT of Peanut

PubMed Central

Gajera, H. P.; Vakharia, D. N.

2012-01-01

Twelve isolates of Trichoderma (six of T. harzianum, five of T. viride, one of T. virens), which reduced variably the incidence of collar rot disease caused in peanut by Aspergillus niger Van Tieghem, were evaluated for their potential to produce lytic enzymes during in vitro antagonism. T. viride 60 inhibited highest (86.2%) growth of test fungus followed by T. harzianum 2J (80.4%) at 6 days after inoculation (DAI) on PDA media. The specific activities of chitinase, β-1,3-glucanase and protease were 11, 3.46 and 9 folds higher in T6 antagonist (T. viride 60 and A. niger interactions) followed by 8.72, 2.85 and 9 folds in T8antagonist (T. harzianum 2J and A. niger interactions), respectively, compared to the activity produced by control petri plate T13 (A. niger alone) at 6 DAI. Activity of these lytic enzymes induced in antagonists’ plates comprises the growth of Trichoderma isolates. However, cellulase and poly galacturonase were found least amount in these antagonists treatment. A significant positive correlation (p=0.01) between percentage growth inhibition of test fungus and lytic enzymes – (chitinase, β-1,3-glucanase and protease) in the culture medium of antagonist treatment established a relationship to inhibit growth of fungal pathogen by increasing the levels of these enzymes. Among the Trichoderma isolates, T. viride 60 was found best strain to be used in biological control of plant pathogen A. niger. PMID:24031802

Overexpression of an aquaglyceroporin gene from Trichoderma harzianum improves water-use efficiency and drought tolerance in Nicotiana tabacum.

PubMed

Vieira, Pabline Marinho; Santos, Mirella Pupo; Andrade, Cristiana Moura; Souza-Neto, Otacílio Antônio; Ulhoa, Cirano José; Aragão, Francisco José Lima

2017-12-01

Aquaporins (AQPs) and aquaglyceroporins (AQGPs) are integral membrane proteins that mediate the transport of water and solutes, such as glycerol and urea, across membranes. AQP and AQGP genes represent a valuable tool for biotechnological improvement of plant tolerance to environmental stresses. We previously isolated a gene encoding for an aquaglyceroporin (ThAQGP), which was up-regulated in Trichoderma harzianum during interaction with the plant pathogen Fusarium solani. This gene was introduced into Nicotiana tabacum and plants were physiologically characterized. Under favorable growth conditions, transgenic progenies did not had differences in both germination and growth rates when compared to wild type. However, physiological responses under drought stress revealed that transgenic plants presented significantly higher transpiration rate, stomatal conductance, photosynthetic efficiency and faster turgor recovery than wild type. Quantitative RT-PCR analysis demonstrated the presence of ThAQGP transcripts in transgenic lines, showing the cause-effect relationship between the observed phenotype and the expression of the transgene. Our results underscore the high potential of T. harzianum as a source of genes with promising applications in transgenic plants tolerant to drought stress. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Can Tomato Inoculation with Trichoderma Compensate Yield and Soil Health Deficiency due to Soil Salinity?

NASA Astrophysics Data System (ADS)

Wagner, Karl; Apostolakis, Antonios; Daliakopoulos, Ioannis; Tsanis, Ioannis

2016-04-01

Soil salinity is a major soil degradation threat, especially for arid coastal environments where it hinders agricultural production and soil health. Protected horticultural crops in the Mediterranean region, typically under deficit irrigation and intensive cultivation practices, have to cope with increasing irrigation water and soil salinization. This study quantifies the beneficial effects of the Trichoderma harzianum (TH) on the sustainable production of Solanum lycopersicum (tomato), a major greenhouse crop of the RECARE project Case Study in Greece, the semi-arid coastal Timpaki basin in south-central Crete. 20 vigorous 20-day-old Solanum lycopersicum L. cv Elpida seedlings are treated with TH fungi (T) or without (N) and transplanted into 35 L pots under greenhouse conditions. Use of local planting soil with initial Electrical Conductivity (ECe) 1.8 dS m-1 and local cultivation practices aim to simulate the prevailing conditions at the Case Study. In order to simulate seawater intrusion affected irrigation, plants are drip irrigated with two NaCl treatments: slightly (S) saline (ECw = 1.1 dS m-1) and moderately (M) saline water (ECw = 3.5 dS m-1), resulting to very high and excessively high ECe, respectively. Preliminary analysis of below and aboveground biomass, soil quality, salinity, and biodiversity indicators, suggest that TH pre-inoculation of tomato plants at both S and M treatments improve yield, soil biodiversity and overall soil health.

De novo transcriptome analyses of host-fungal interactions in oil palm (Elaeis guineensis Jacq.).

PubMed

Ho, Chai-Ling; Tan, Yung-Chie; Yeoh, Keat-Ai; Ghazali, Ahmad-Kamal; Yee, Wai-Yan; Hoh, Chee-Choong

2016-01-19

Basal stem rot (BSR) is a fungal disease in oil palm (Elaeis guineensis Jacq.) which is caused by hemibiotrophic white rot fungi belonging to the Ganoderma genus. Molecular responses of oil palm to these pathogens are not well known although this information is crucial to strategize effective measures to eradicate BSR. In order to elucidate the molecular interactions between oil palm and G. boninense and its biocontrol fungus Trichoderma harzianum, we compared the root transcriptomes of untreated oil palm seedlings with those inoculated with G. boninense and T. harzianum, respectively. Differential gene expression analyses revealed that jasmonate (JA) and salicylate (SA) may act in an antagonistic manner in affecting the hormone biosynthesis, signaling, and downstream defense responses in G. boninense-treated oil palm roots. In addition, G. boninense may compete with the host to control disease symptom through the transcriptional regulation of ethylene (ET) biosynthesis, reactive oxygen species (ROS) production and scavenging. The strengthening of host cell walls and production of pathogenesis-related proteins as well as antifungal secondary metabolites in host plants, are among the important defense mechanisms deployed by oil palm against G. boninense. Meanwhile, endophytic T. harzianum was shown to improve the of nutrition status and nutrient transportation in host plants. The findings of this analysis have enhanced our understanding on the molecular interactions of G. boninense and oil palm, and also the biocontrol mechanisms involving T. harzianum, thus contributing to future formulations of better strategies for prevention and treatment of BSR.

Isolation and identification of Trichoderma harzianum from groundwater: An effective biosorbent for defluoridation of groundwater.

PubMed

Koshle, Shalini; Mahesh, S; Swamy, S Nanjunda

2016-01-01

The ability of non-viable form of Trichoderma harzianum, isolated from fluoride rich groundwater, was investigated as biosorbent for defluoridation of groundwater. Biosorption experiments were carried out at laboratory scale for removal of fluoride from groundwater. Significant effect of operational parameters on fluoride biosorption using Trichoderma harzianum as biosorbent was evaluated by varying operational parameters such as: initial fluoride concentration (2-8 mgl(-1)), biosorbent dose (0.4-1.6g/100ml), groundwater pH (6-10), temperature (30-50 degrees C) and biosorption time (30-120 min). The fluoride adsorption isotherms were modeled by Langmuir and Freundlich isotherms. Our result showed that fluoride biosorption, significantly increased with increase in groundwater pH, biosorbent dose, temperature and biosorption time, whereas increase in initial fluoride concentration reduced fluoride removal. The fluoride biosorption was rapid and maximum fluoride uptake was attained with 1.6g 100ml(-1) biosorbent within 60 min. Optimal pH 10 and temperature 50 degrees C gave maximum defluoridation efficiency. Freundlich isotherm fits well for defluoridation of groundwater using Trichoderma harzianum as biosorbent which indicated that biosorbent surface sites were heterogeneous in nature and fitted into heterogeneous site binding model.

Progress towards mapping the constitutional t(11:22) breakpoint

DOE Office of Scientific and Technical Information (OSTI.GOV)

Barnoski, B.L.; Emanuel, B.S.; Bell, C.J.

1994-09-01

The reciprocal t(11;22)(q23;q11) is the most frequent, recurrent, non-Robertsonian, constitutional translocation in humans. Balanced carriers of this rearrangement are phenotypically normal, but are at risk for producing abnormal offspring with the Supernumerary der(22)t(11;22) Syndrome. Further, a recent report of association between t(11;22) balanced translocation carriers and breast cancer, suggests the involvement of genes on 11q and/or 22q in breast cancer tumorigenesis. Studies are in progress to examine the similarity between 11q23 and 22q11 breakpoints in multiple families with the constitutional t(11;22). A 750 kb YAC, which contains markers known to flank the 11q23 breakpoint, was identified in CEPH/Genethon database. FISHmore » with this YAC to two independent t(11;22) cell lines demonstrates signal on both derivative chromosomes. Numerous YACs containing BCRL2, the closest marker proximal to the breakpoint, were identified. Analysis of these YACs to determine which contain the actual breakpoint sequences is complicated by the presence of a duplicated segment of 22q11 which contains a GGTL and a BCRL locus. Sequences homologous to these loci are present at several other locations in 22q11. The BCRL positive YACs were analyzed by Southern hybridization under conditions which distinguish the four members of the BCR/BCRL family. FISH of total yeast DNA plus YAC DNA labeled by nick translation, or biotin-labeled inter-Alu PCR products confirmed the localization of these YACs to 22q11. Additional FISH with these YACS to metaphase spreads prepared from balanced t(11;22) carriers confirm that these clones span the breakpoint, and will allow rapid isolation and definition of the genetic region adjacent to the t(11;22) breakpoint.« less

Use of a new Trichoderma harzianum strain isolated from the Amazon rainforest with pretreated sugar cane bagasse for on-site cellulase production.

PubMed

Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; da Silva, Mateus Ribeiro; Azzoni, Sindelia Freitas; Pradella, José Geraldo da Cruz

2012-03-01

The on-site production of cellulases is an important strategy for the development of sustainable second-generation ethanol production processes. This study concerns the use of a specific cellulolytic enzyme complex for hydrolysis of pretreated sugar cane bagasse. Glycosyl hydrolases (FPase, xylanase, and β-glucosidase) were produced using a new strain of Trichoderma harzianum, isolated from the Amazon rainforest and cultivated under different conditions. The influence of the carbon source was first investigated using shake-flask cultures. Selected carbon sources were then further studied under different pH conditions using a stirred tank bioreactor. Enzymatic activities up to 121 FPU/g, 8000 IU/g, and 1730 IU/g of delignified steam-exploded bagasse+sucrose were achieved for cellulase, xylanase and β-glucosidase, respectively. This enzymatic complex was used to hydrolyze pretreated sugar cane bagasse. A comparative evaluation, using an enzymatic extract from Trichoderma reesei RUTC30, indicated similar performance of the T. harzianum enzyme complex, being a potential candidate for on-site production of enzymes. Copyright © 2011 Elsevier Ltd. All rights reserved.

Effect of Paecilomyces lilacinus, Trichoderma harzianum and Trichoderma virens fungal extracts on the hatchability of Ancylostoma eggs.

PubMed

Hofstätter, Bianca Delgado Menezes; Oliveira da Silva Fonseca, Anelise; de Souza Maia Filho, Fernando; de Souza Silveira, Julia; Persici, Beatriz Maroneze; Pötter, Luciana; Silveira, Andressa; Antoniolli, Zaida Inês; Brayer Pereira, Daniela Isabel

Ancylostoma species have demanded attention due to their zoonotic potential. The use of anthelmintics is the usual method to prevent environmental contamination by Ancylostoma eggs and larvae. Nematophagous fungi have been widely used in their biological control due to the fungus ability to capture and digest free nematode forms. The aim of this study was to evaluate the effect of four different fungal extracts of Paecilomyces lilacinus (n=2), Trichoderma harzianum (n=1) and Trichoderma virens (n=1) isolates on the hatchability of Ancylostoma eggs. Fungal extracts consisted of fungal broth culture supernatant without filtration (crude extract) and filtered broth (filtered extract), macerated mycelium (crude macerate), and macerated mycelium submitted to filtration (filtered macerate). The Ancylostoma eggs were obtained from the feces of naturally infected dogs. In vitro assays were performed in five replicates and consisted of four treatments and one control group. The activity of the fungal extracts of each evaluated fungus differed (p<0.05) from those of the control group, showing significant ovicidal activity. The hatching of the eggs suffered reduction percentages of 68.43% and 47.05% with P. lilacinus, and 56.43% with T. harzianum, when crude macerate extract was used. The reduction with the macerate extract of T. virens was slightly lower (52.25%) than that for the filtered macerate (53.64%). The results showed that all extracts were effective in reducing the hatchability of Ancylostoma eggs. The ovicidal effect observed is likely to have been caused by the action of hydrolytic enzymes secreted by the fungi. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

Salicylic acid prevents Trichoderma harzianum from entering the vascular system of roots.

PubMed

Alonso-Ramírez, Ana; Poveda, Jorge; Martín, Ignacio; Hermosa, Rosa; Monte, Enrique; Nicolás, Carlos

2014-10-01

Trichoderma is a soil-borne fungal genus that includes species with a significant impact on agriculture and industrial processes. Some Trichoderma strains exert beneficial effects in plants through root colonization, although little is known about how this interaction takes place. To better understand this process, the root colonization of wild-type Arabidopsis and the salicylic acid (SA)-impaired mutant sid2 by a green fluorescent protein (GFP)-marked Trichoderma harzianum strain was followed under confocal microscopy. Trichoderma harzianum GFP22 was able to penetrate the vascular tissue of the sid2 mutant because of the absence of callose deposition in the cell wall of root cells. In addition, a higher colonization of sid2 roots by GFP22 compared with that in Arabidopsis wild-type roots was detected by real-time polymerase chain reaction. These results, together with differences in the expression levels of plant defence genes in the roots of both interactions, support a key role for SA in Trichoderma early root colonization stages. We observed that, without the support of SA, plants were unable to prevent the arrival of the fungus in the vascular system and its spread into aerial parts, leading to later collapse. © 2014 BSPP AND JOHN WILEY & SONS LTD.

In vitro antagonism of cotton seedlings fungi and characterization of chitinase isozyme activities in Trichoderma harzianum.

PubMed

Asran-Amal, A; Moustafa-Mahmoud, S M; Sabet, K K; El Banna, O H

2010-04-01

The antagonistic fungus Trichoderma harzianum is widely recognized as a potential biocontrol agent against several soil-borne plant pathogens. T. harzinum is rich source of chitinoltic enzymes. In vitro screening of 5 isolates of T. harzinum, one isolate of Chaetomium globosum and one isolate of Conetherium mentance, revealed that all of them had reduced growth area of Macrophomina phaseolina, Fusarium solani and Rhizoctonia solani on PDA medium, significantly. The inhibition percentage ranged from 77.9 % to 55.9% for M. phaseolina and 59.2% to 40.4% for R. solani by T. harzinum and C. mentance, respectively. Inhibition for F. solani ranged from 76.5% to 55.7% by T. harzinum and C. globosum, respectively. Isozyme gel electrophoresis was used to assess chitinase activity secreted by selected isolates of T. harzinum under different pH degrees and temperatures. Obtained results indicated that activity of chitinase isozyme produced at 30 °C was higher than 15-20 °C for all tested isolates and activity of chitinase produced by isolates No. 4 and 5 of T. harzinum at pH (7-7.5) was higher than at pH 6, respectively.

Differential Response of Extracellular Proteases of Trichoderma Harzianum Against Fungal Phytopathogens.

PubMed

Sharma, Vivek; Salwan, Richa; Sharma, Prem N

2016-09-01

In the present study, production of extracellular proteases by Trichoderma harzianum was evaluated based on the relative gene expression and spectrophotometric assay. The fungal isolates were grown in Czapek Dox Broth medium supplemented with deactivated mycelium of plant fungal pathogens such as Fusarium oxysporum, Colletotrichum capsici, Gloeocercospora sorghi, and Colletotrichum truncatum. The maximum protease activity was detected after 48 h of incubation against Colletotrichum spp. Similarly in qRT-PCR, the relative gene expression of four proteases varied from 48 to 96 h against host pathogens in a time-independent manner. Among proteases, statistically significant upregulation of asp, asp, and srp was observed against Colletotrichum spp., followed by F. oxysporum. But in the case of pepM22, maximum upregulation was observed against F. oxysporum. The variation in enzyme assay and qRT-PCR of proteases at different time intervals against various fungal phytopathogens could be due to the limitation of using casein as a substrate for all types of proteases or protease-encoding transcripts selected for qRT-PCR, which may not be true representative of total protease activity.

Purification and characterization of an endo-beta-1,6-glucanase from Trichoderma harzianum that is related to its mycoparasitism.

PubMed Central

de la Cruz, J; Pintor-Toro, J A; Benítez, T; Llobell, A

1995-01-01

The enzymes from Trichoderma species that degrade fungal cell walls have been suggested to play an important role in mycoparasitic action against fungal plant pathogens. The mycoparasite Trichoderma harzianum produces at least two extracellular beta-1,6-glucanases, among other hydrolases, when it is grown on chitin as the sole carbon source. One of these extracellular enzymes was purified to homogeneity after adsorption to its substrate, pustulan, chromatofocusing, and, finally, gel filtration. The apparent molecular mass was 43,000, and the isoelectric point was 5.8. The first 15 amino acids from the N terminus of the purified protein have been sequenced. The enzyme was specific for beta-1,6 linkages and showed an endolytic mode of action on pustulan. Further characterization indicated that the enzyme by itself releases soluble sugars and produces hydrolytic halli on yeast cell walls. When combined with other T. harzianum cell wall-degrading enzymes such as beta-1,3-glucanases and chitinases, it hydrolyzes filamentous fungal cell walls. The enzyme acts cooperatively with the latter enzymes, inhibiting the growth of the fungi tested. Antibodies against the purified protein also indicated that the two identified beta-1,6-glucanases are not immunologically related and are probably encoded by two different genes. PMID:7896713

Quantification of the biocontrol agent Trichoderma harzianum with real-time TaqMan PCR and its potential extrapolation to the hyphal biomass.

PubMed

López-Mondéjar, Rubén; Antón, Anabel; Raidl, Stefan; Ros, Margarita; Pascual, José Antonio

2010-04-01

The species of the genus Trichoderma are used successfully as biocontrol agents against a wide range of phytopathogenic fungi. Among them, Trichoderma harzianum is especially effective. However, to develop more effective fungal biocontrol strategies in organic substrates and soil, tools for monitoring the control agents are required. Real-time PCR is potentially an effective tool for the quantification of fungi in environmental samples. The aim of this study consisted of the development and application of a real-time PCR-based method to the quantification of T. harzianum, and the extrapolation of these data to fungal biomass values. A set of primers and a TaqMan probe for the ITS region of the fungal genome were designed and tested, and amplification was correlated to biomass measurements obtained with optical microscopy and image analysis, of the hyphal length of the mycelium of the colony. A correlation of 0.76 between ITS copies and biomass was obtained. The extrapolation of the quantity of ITS copies, calculated based on real-time PCR data, into quantities of fungal biomass provides potentially a more accurate value of the quantity of soil fungi. Copyright 2009 Elsevier Ltd. All rights reserved.

Trichoharzianol, a new antifungal from Trichoderma harzianum F031.

PubMed

Jeerapong, Chotika; Phupong, Worrapong; Bangrak, Phuwadol; Intana, Warin; Tuchinda, Patoomratana

2015-04-15

A new decalin derivative, trichoharzianol (1), together with three known compounds, eujavanicol A (2), 5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone (3), and 4,6-dihydroxy-5-methylphthalide (4), were isolated from Trichoderma harzianum F031. For the first time, compounds 2-4 were reported from the Trichoderma species. Their structures were characterized by spectroscopic methods. Trichoharzianol (1) showed the highest antifungal activity against Colletotrichum gloeosporioides, with a minimum inhibitory concentration (MIC) of 128 μg/mL.

Impact of a native Streptomyces flavovirens from mushroom compost on green mold control and yield of Agaricus bisporus.

PubMed

Šantrić, Ljiljana; Potočnik, Ivana; Radivojević, Ljiljana; Umiljendić, Jelena Gajić; Rekanović, Emil; Duduk, Bojan; Milijašević-Marčić, Svetlana

2018-05-18

Thirty-five actinobacterial isolates, obtained from button mushroom (Agaricus bisporus) substrates (i.e., compost in different phases of composting, black peat or casing layer) in Serbia in 2014-2016 were tested in vitro against the causal agents of green mold in cultivated mushroom. Out of six most promising isolates, A06 induced 42.4% in vitro growth inhibition of Trichoderma harzianum T54, and 27.6% inhibition of T. aggressivum f. europaeum T77. The novel strain A06 was identified as Streptomyces flavovirens based on macroscopic and cultural characteristics and 16S rDNA sequence and used in mushroom growing room experiments. Actinobacteria had no negative influence on mycelial growth of the cultivated mushroom in compost in situ. Isolate S. flavovirens A06 enhanced mushroom yield significantly, up to 31.5%. The A06 isolate was more efficient in enhancing yield after inoculation with the compost mold T. aggressivum (26.1%), compared to casing mold T. harzianum (8%). Considering disease incidence, actinobacteria significantly prevented green mold in compost caused by T. aggressivum (6.8%). However, fungicide prochloraz-Mn had a more significant role in reducing symptoms of casing mold, T. harzianum, in comparison with actinobacteria (24.2 and 11.8%, respectively). No significant differences between efficacies of S. flavovirens A06 and the fungicide prochloraz-Mn against T. aggressivum were revealed. These results imply that S. flavovirens A06 can be used to increase mushroom yield and contribute to disease control against the aggressive compost green mold disease caused by Trichoderma aggressivum.

Improvement of growth of Eucalyptus globulus and soil biological parameters by amendment with sewage sludge and inoculation with arbuscular mycorrhizal and saprobe fungi.

PubMed

Arriagada, C; Sampedro, I; Garcia-Romera, I; Ocampo, J

2009-08-15

Sewage sludge is widely used as an organic soil amendment to improve soil fertility. We investigated the effects of sewage sludge (SS) application on certain biological parameters of Eucalyptus globulus Labill. The plant was either uninoculated or inoculated with saprobe fungi (Coriolopsis rigida and Trichoderma harzianum) or arbuscular mycorrhizal (AM) fungi (Glomus deserticola and Gigaspora rosea). Sewage sludge was applied to the surface of experimental plots at rates of 0, 2, 4, 6 and 8 g 100 g(-1) of soil. Inoculation with both AM and saprobe fungi in the presence of SS was essential for the promotion of plant growth. The AM, saprobe fungi and SS significantly increased dry shoot weight. The AM fungi induced a significant increase in Fluorescein diacetate (FDA) activity but did not increase beta-glucosidase activity. Addition of SS to AM-inoculated soil did not affect either FDA or alpha-glucosidase activities in plants from soil that was either uninoculated or inoculated with the saprobe fungi. SS increased beta-glucosidase activity when it was applied at 4 g 100 g(-1). SS negatively affected AM colonization as well as the mycelium SDH activity for both mycorrhizal fungi. SS increased Eucalyptus shoot biomass and enhanced its nutrient status. Inoculation of the soil with G. deserticola stimulated significant E. globulus growth and increases in shoot tissue content of N, P, K, Ca, Mg and Fe. Dual inoculation with G. deserticola and either of the saprobe fungi had positive effects on K, Ca, Mg and Fe contents. The application of 8 g 100 g(-1) of SS had no positive effects on plant nutrition. The experimental setup provided a suitable tool for evaluating SS in combination with saprobe and AM fungi as a biological fertiliser for its beneficial effects on E. globulus plant growth.

An integrated approach for the reduction of aflatoxin contamination in chilli (Capsicum annuum L.).

PubMed

Sudha, S; Naik, M K; Ajithkumar, K

2013-02-01

An integrated approach for management of aflatoxin contamination in chilli was undertaken by evaluating the fungicides, bioagents and plant extracts against Aspergillus flavus under both in vitro and field condition. Maximum inhibition of radial growth (91.1%) was observed with 0.3% mancozeb followed by captan (85.2%). Carbendazim (73%) was effective and superior over other systemic fungicides. A complete inhibition (100%) of A. flavus was observed in neem seed kernel extract (NSKE), nimbicidin and pongamia oil at 5%. An indigenous Pseudomonas fluorescens bioagent isolate inhibited (74.9%) the growth of A. flavus over Trichoderma harzianum (70.4%). The superior performing fungicides, plant extracts and bioagents identified under in vitro were used for challenge inoculation on chilli fruits and so also for field evaluation. The captan treated fruits recorded the least infection of A. flavus (1.6%) followed by P. fluorescens (2.0%), NSKE (2.2%) and nimbicidin treated fruits (7.8%) as against control (38.3%). As regards to field evaluation, the least incidence was recorded in NSKE sprayed chilli plot (1.6%) and was on par with captan (2.2%), P. fluorescens (2.4%) and T. harzianum (2.6%) compared to control (7.4%). Hence, a pre-harvest spray of NSKE (5%) or mancozeb (0.3%) or P. fluorescens (1 × 10(8) cfu/ml) 10 days before harvest of chilli is recommended for field level management of aflatoxin.

Production of 6-pentyl-α-pyrone by trichoderma harzianum in solid-state fermentation

PubMed Central

de Souza Ramos, Aline; Fiaux, Sorele Batista; Leite, Selma Gomes Ferreira

2008-01-01

Many Trichoderma species are able to produce 6-pentyl-α-pyrone (6-PP), a lactone with coconut-like aroma. In the present work, several culture parameters were studied to enhance the production of 6-PP by Trichoderma harzianum 4040 in solid-state fermentation. Green coir powder added to a nutrient solution was used as support material for fermentation. A Plackett-Burman screening technique was applied, followed by a fractionary factorial design. The best culture conditions within the experimental domain studied were (100 g support)−1: sucrose, 3 g; NaNO3, 0.24 g; (NH4)2SO4, 0.18 g; KH2PO4, 0.1 g; inoculum concentration, 2.2 × 106 spores; moisture level, 55%. The temperature established was 28°C. The fermentation under the selected conditions led to a 6-PP production six times higher (5.0 mg/g dry matter) than the initial one (0.8 mg/g dry matter) after seven days of cultivation. PMID:24031295

Production of 6-pentyl-α-pyrone by trichoderma harzianum in solid-state fermentation.

PubMed

de Souza Ramos, Aline; Fiaux, Sorele Batista; Leite, Selma Gomes Ferreira

2008-10-01

Many Trichoderma species are able to produce 6-pentyl-α-pyrone (6-PP), a lactone with coconut-like aroma. In the present work, several culture parameters were studied to enhance the production of 6-PP by Trichoderma harzianum 4040 in solid-state fermentation. Green coir powder added to a nutrient solution was used as support material for fermentation. A Plackett-Burman screening technique was applied, followed by a fractionary factorial design. The best culture conditions within the experimental domain studied were (100 g support)(-1): sucrose, 3 g; NaNO3, 0.24 g; (NH4)2SO4, 0.18 g; KH2PO4, 0.1 g; inoculum concentration, 2.2 × 10(6) spores; moisture level, 55%. The temperature established was 28°C. The fermentation under the selected conditions led to a 6-PP production six times higher (5.0 mg/g dry matter) than the initial one (0.8 mg/g dry matter) after seven days of cultivation.

Deciphering the hormonal signalling network behind the systemic resistance induced by Trichoderma harzianum in tomato

PubMed Central

Martínez-Medina, Ainhoa; Fernández, Iván; Sánchez-Guzmán, María J.; Jung, Sabine C.; Pascual, Jose A.; Pozo, María J.

2013-01-01

Root colonization by selected Trichoderma isolates can activate in the plant a systemic defense response that is effective against a broad-spectrum of plant pathogens. Diverse plant hormones play pivotal roles in the regulation of the defense signaling network that leads to the induction of systemic resistance triggered by beneficial organisms [induced systemic resistance (ISR)]. Among them, jasmonic acid (JA) and ethylene (ET) signaling pathways are generally essential for ISR. However, Trichoderma ISR (TISR) is believed to involve a wider variety of signaling routes, interconnected in a complex network of cross-communicating hormone pathways. Using tomato as a model, an integrative analysis of the main mechanisms involved in the systemic resistance induced by Trichoderma harzianum against the necrotrophic leaf pathogen Botrytis cinerea was performed. Root colonization by T. harzianum rendered the leaves more resistant to B. cinerea independently of major effects on plant nutrition. The analysis of disease development in shoots of tomato mutant lines impaired in the synthesis of the key defense-related hormones JA, ET, salicylic acid (SA), and abscisic acid (ABA), and the peptide prosystemin (PS) evidenced the requirement of intact JA, SA, and ABA signaling pathways for a functional TISR. Expression analysis of several hormone-related marker genes point to the role of priming for enhanced JA-dependent defense responses upon pathogen infection. Together, our results indicate that although TISR induced in tomato against necrotrophs is mainly based on boosted JA-dependent responses, the pathways regulated by the plant hormones SA- and ABA are also required for successful TISR development. PMID:23805146

Trichoderma-Inoculated Miscanthus Straw Can Replace Peat in Strawberry Cultivation, with Beneficial Effects on Disease Control.

PubMed

Debode, Jane; De Tender, Caroline; Cremelie, Pieter; Lee, Ana S; Kyndt, Tina; Muylle, Hilde; De Swaef, Tom; Vandecasteele, Bart

2018-01-01

Peat based growing media are not ecologically sustainable and often fail to support biological control. Miscanthus straw was (1) tested to partially replace peat; and (2) pre-colonized with a Trichoderma strain to increase the biological control capacity of the growing media. In two strawberry pot trials (denoted as experiment I & II), extruded and non-extruded miscanthus straw, with or without pre-colonization with T. harzianum T22, was used to partially (20% v/v) replace peat. We tested the performance of each mixture by monitoring strawberry plant development, nutrient content in the leaves and growing media, sensitivity of the fruit to the fungal pathogen Botrytis cinerea , rhizosphere community and strawberry defense responses. N immobilization by miscanthus straw reduced strawberry growth and yield in experiment II but not in I. The pre-colonization of the straw with Trichoderma increased the post-harvest disease suppressiveness against B. cinerea and changed the rhizosphere fungal microbiome in both experiments. In addition, defense-related genes were induced in experiment II. The use of miscanthus straw in growing media will reduce the demand for peat and close resource loops. Successful pre-colonization of this straw with biological control fungi will optimize crop cultivation, requiring fewer pesticide applications, which will benefit the environment and human health.

Test wells T21, T22, and T25, White Sands Missile Range, Dona Ana County, New Mexico

USGS Publications Warehouse

Myers, R.G.

1983-01-01

Three test wells, T21, T22, and T25, were drilled at White Sands Missile Range in south-central New Mexico as part of a joint military program sponsored by the U.S. Army in September 1982. T21 and T22 were drilled as observation wells for two old landfills. T25 was drilled as an exploratory hole to obtain lithologic and borehole-geophysical data in the vicinity of the proposed replacement well for Supply Well 15. Information obtained from these wells includes borehole-geophysical and driller's logs.

Trichoderma harzianum elicits induced resistance in sunflower challenged by Rhizoctonia solani.

PubMed

Singh, B N; Singh, A; Singh, B R; Singh, H B

2014-03-01

To investigate the efficacy of Trichoderma harzianum NBRI-1055 (denoted as 'T-1055') in suppression of seedling blight of sunflower caused by Rhizoctonia solani Kühn and their impact on host defence responses. T-1055 was applied as seed treatment, soil application and combined application (seed treatment + soil application). Higher protection afforded by combined application of T-1055 was associated with the marked induction of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), peroxidase (PO) and cinnamyl alcohol dehydrogenase (CAD) activities. The activities of PAL and PPO reached maximum at 10 days after sowing (DAS), while PO and CAD levels reached maximum at 12 DAS. This was further supported by the accumulation of total phenolic content that showed an increase up to threefold at 14 DAS. In addition, HPLC analysis revealed that the contents of ferulic and p-coumaric acids increased by 6·3 and 4·6 times, respectively, at 14 DAS. Amount of gallic acid was also little more than double. Lignin deposition in sunflower root increased by 2·7, 3·4 and 3·7 times through combined application of T-1055 at 16, 18 and 20 DAS, respectively. Combined application also increased the accumulation of PR-2 and PR-3 proteins by 3·3 and 3·9 times, respectively, at 12 DAS in followed by seed treatment alone. The combined application of T-1055 triggered defence responses in an enhanced level in sunflower than the soil and seed alone and provided better protection against Rhizoctonia seedling blight. Rhizospheric fungal bioagent 'T-1055' can enhance protection in sunflower against the R. solani pathogen through augmented elicitation of host defence responses. © 2013 The Society for Applied Microbiology.

Bacterial Compatibility in Combined Inoculations Enhances the Growth of Potato Seedlings.

PubMed

Santiago, Christine D; Yagi, Shogo; Ijima, Motoaki; Nashimoto, Tomoya; Sawada, Maki; Ikeda, Seishi; Asano, Kenji; Orikasa, Yoshitake; Ohwada, Takuji

2017-03-31

The compatibility of strains is crucial for formulating bioinoculants that promote plant growth. We herein assessed the compatibility of four potential bioinoculants isolated from potato roots and tubers (Sphingomonas sp. T168, Streptomyces sp. R170, Streptomyces sp. R181, and Methylibium sp. R182) that were co-inoculated in order to improve plant growth. We screened these strains using biochemical tests, and the results obtained showed that R170 had the highest potential as a bioinoculant, as indicated by its significant ability to produce plant growth-promoting substances, its higher tolerance against NaCl (2%) and AlCl 3 (0.01%), and growth in a wider range of pH values (5.0-10.0) than the other three strains. Therefore, the compatibility of R170 with other strains was tested in combined inoculations, and the results showed that the co-inoculation of R170 with T168 or R182 synergistically increased plant weight over un-inoculated controls, indicating the compatibility of strains based on the increased production of plant growth promoters such as indole-3-acetic acid (IAA) and siderophores as well as co-localization on roots. However, a parallel test using strain R181, which is the same Streptomyces genus as R170, showed incompatibility with T168 and R182, as revealed by weaker plant growth promotion and a lack of co-localization. Collectively, our results suggest that compatibility among bacterial inoculants is important for efficient plant growth promotion, and that R170 has potential as a useful bioinoculant, particularly in combined inoculations that contain compatible bacteria.

Production of a recombinant swollenin from Trichoderma harzianum in Escherichia coli and its potential synergistic role in biomass degradation.

PubMed

Santos, Clelton A; Ferreira-Filho, Jaire A; O'Donovan, Anthonia; Gupta, Vijai K; Tuohy, Maria G; Souza, Anete P

2017-05-16

Fungal swollenins (SWOs) constitute a class of accessory proteins that are homologous to canonical plant expansins. Expansins and expansin-related proteins are well known for acting in the deagglomeration of cellulose structure by loosening macrofibrils. Consequently, SWOs can increase the accessibility and efficiency of the other enzymes involved in the saccharification of cellulosic substrates. Thus, SWOs are promising targets for improving the hydrolysis of plant biomass and for use as an additive to enhance the efficiency of an enzyme cocktail designed for the production of biofuels. Here, we report the initial characterization of an SWO from Trichoderma harzianum (ThSwo) that was successfully produced using Escherichia coli as a host. Initially, transcriptome and secretome data were used to compare swo gene expression and the amount of secreted ThSwo. The results from structural modeling and phylogenetic analysis of the ThSwo protein showed that ThSwo does preserve some structural features of the plant expansins and family-45 glycosyl hydrolase enzymes, but it evolutionarily diverges from both of these protein classes. Recombinant ThSwo was purified at a high yield and with high purity and showed secondary folding similar to that of a native fungal SWO. Bioactivity assays revealed that the purified recombinant ThSwo created a rough and amorphous surface on Avicel and displayed a high synergistic effect with a commercial xylanase from T. viride, enhancing its hydrolytic performance up to 147 ± 7%. Many aspects of the structure and mechanism of action of fungal SWOs remain unknown. In the present study, we produced a recombinant, active SWO from T. harzianum using a prokaryotic host and confirmed its potential synergistic role in biomass degradation. Our work paves the way for further studies evaluating the structure and function of this protein, especially regarding its use in biotechnology.

Comparative studies on the induction of Trichoderma harzianum mutanase by α-(1→3)-glucan-rich fruiting bodies and mycelia of Laetiporus sulphureus.

PubMed

Wiater, Adrian; Pleszczyńska, Małgorzata; Szczodrak, Janusz; Janusz, Grzegorz

2012-01-01

Mutanase (α-(1→3)-glucanase) is a little-known inductive enzyme that is potentially useful in dentistry. Here, it was shown that the cell wall preparation (CWP) obtained from the fruiting body or vegetative mycelium of polypore fungus Laetiporus sulphureus is rich in α-(1→3)-glucan and can be successfully used for mutanase induction in Trichoderma harzianum. The content of this biopolymer in the CWP depended on the age of fruiting bodies and increased along with their maturation. In the case of CWP prepared from vegetative mycelia, the amount of α-(1→3)-glucan depended on the mycelium age and also on the kind of medium used for its cultivation. All CWPs prepared from the individually harvested fruiting body specimens induced high mutanase activity (0.53-0.82 U/mL) in T. harzianum after 3 days of cultivation. As for the CWPs obtained from the hyphal mycelia of L. sulpureus, the maximal enzyme productivity (0.34 U/mL after 3 days of incubation) was recorded for CWP prepared from the 3 week-old mycelium cultivated in Sabouraud medium. Statistically, a high positive correlation was found between the total percentage content of α-(1→3)-glucan in the CWP and the mutanase activity.

Colonization of onions by endophytic fungi and their impacts on the biology of Thrips tabaci.

PubMed

Muvea, Alexander M; Meyhöfer, Rainer; Subramanian, Sevgan; Poehling, Hans-Michael; Ekesi, Sunday; Maniania, Nguya K

2014-01-01

Endophytic fungi, which live within host plant tissues without causing any visible symptom of infection, are important mutualists that mediate plant-herbivore interactions. Thrips tabaci (Lindeman) is one of the key pests of onion, Allium cepa L., an economically important agricultural crop cultivated worldwide. However, information on endophyte colonization of onions, and their impacts on the biology of thrips feeding on them, is lacking. We tested the colonization of onion plants by selected fungal endophyte isolates using two inoculation methods. The effects of inoculated endophytes on T. tabaci infesting onion were also examined. Seven fungal endophytes used in our study were able to colonize onion plants either by the seed or seedling inoculation methods. Seed inoculation resulted in 1.47 times higher mean percentage post-inoculation recovery of all the endophytes tested as compared to seedling inoculation. Fewer thrips were observed on plants inoculated with Clonostachys rosea ICIPE 707, Trichoderma asperellum M2RT4, Trichoderma atroviride ICIPE 710, Trichoderma harzianum 709, Hypocrea lixii F3ST1 and Fusarium sp. ICIPE 712 isolates as compared to those inoculated with Fusarium sp. ICIPE 717 and the control treatments. Onion plants colonized by C. rosea ICIPE 707, T. asperellum M2RT4, T. atroviride ICIPE 710 and H. lixii F3ST1 had significantly lower feeding punctures as compared to the other treatments. Among the isolates tested, the lowest numbers of eggs were laid by T. tabaci on H. lixii F3ST1 and C. rosea ICIPE 707 inoculated plants. These results extend the knowledge on colonization of onions by fungal endophytes and their effects on Thrips tabaci.

Colonization of Onions by Endophytic Fungi and Their Impacts on the Biology of Thrips tabaci

PubMed Central

Muvea, Alexander M.; Meyhöfer, Rainer; Subramanian, Sevgan; Poehling, Hans-Michael; Ekesi, Sunday; Maniania, Nguya K.

2014-01-01

Endophytic fungi, which live within host plant tissues without causing any visible symptom of infection, are important mutualists that mediate plant–herbivore interactions. Thrips tabaci (Lindeman) is one of the key pests of onion, Allium cepa L., an economically important agricultural crop cultivated worldwide. However, information on endophyte colonization of onions, and their impacts on the biology of thrips feeding on them, is lacking. We tested the colonization of onion plants by selected fungal endophyte isolates using two inoculation methods. The effects of inoculated endophytes on T. tabaci infesting onion were also examined. Seven fungal endophytes used in our study were able to colonize onion plants either by the seed or seedling inoculation methods. Seed inoculation resulted in 1.47 times higher mean percentage post-inoculation recovery of all the endophytes tested as compared to seedling inoculation. Fewer thrips were observed on plants inoculated with Clonostachys rosea ICIPE 707, Trichoderma asperellum M2RT4, Trichoderma atroviride ICIPE 710, Trichoderma harzianum 709, Hypocrea lixii F3ST1 and Fusarium sp. ICIPE 712 isolates as compared to those inoculated with Fusarium sp. ICIPE 717 and the control treatments. Onion plants colonized by C. rosea ICIPE 707, T. asperellum M2RT4, T. atroviride ICIPE 710 and H. lixii F3ST1 had significantly lower feeding punctures as compared to the other treatments. Among the isolates tested, the lowest numbers of eggs were laid by T. tabaci on H. lixii F3ST1 and C. rosea ICIPE 707 inoculated plants. These results extend the knowledge on colonization of onions by fungal endophytes and their effects on Thrips tabaci. PMID:25254657

Attempts to control Fusarium root rot of bean by seed dressing.

PubMed

Gilardi, G; Baudino, M; Gullino, M L; Garibaldi, A

2008-01-01

In summer 2006, a root rot caused by Fusarium oxysporum was observed in commercial farms on common bean (Phaseolus vulgaris) on the cv Billò and Borlotto. A study was undertaken in order to evaluate the efficacy of different biological control agents applied as seed dressing. In the presence of a medium-high disease incidence, among the biocontrol agents tested, Trichoderma harzianum T 22, Bacillus subtilis QST 713, followed by Pseudomonas chlororaphis, provided generally the best control. Their efficacy was also consistent in the different trials. Also the mixture of T. harzianum + T. viride provide a good disease control. Streptomyces griseoviridis and the 3 strains of Fusarim oxysporum, although less effective, provided a partial control of the disease. The fungicide mancozeb provided only a partial disease control.

Effect of mycosynthesized silver nanoparticles from filtrate of Trichoderma harzianum against larvae and pupa of dengue vector Aedes aegypti L.

PubMed

Sundaravadivelan, Chandran; Padmanabhan, Madanagopal Nalini

2014-03-01

Mosquitoes transmit dreadful diseases, causing millions of deaths every year. Therefore, screening for larvicidal and pupicidal activity of microbial extracts attributes could lead to development of new and improved mosquito control methods that are economical and safe for nontarget organisms and are ecofriendly. Synthetic chemical insecticides occupy predominant position in control strategies. These hazardous chemicals exert unwarranted toxicity and lethal effects on nontarget organisms, develop physiological resistance in target, and cause adverse environmental effect. For vector control, fungal-mediated natural products have been a priority in this area at present. In the current study, effective larvicidal and pupicidal effect of mycosynthesized silver nanoparticles (Ag NPs) using an entomopathogenic fungi Trichoderma harzianum against developmental stages of the dengue vector Aedes aegypti was investigated. An attractive possibility of green nanotechnology is to use microorganisms in the synthesis of nanosilver especially Ag NPs. The mycosynthesized Ag NPs were characterized to find their unique properties through UV-visible spectrophotometer, X-ray diffraction analysis, Fourier transform infrared, and surface characteristics by scanning electron microscopy. To analyze the bioefficacy, different test concentrations for extracellular filtrate (0.2, 0.4, 0.6, 0.8, and 1.0 %) and Ag NPs (0.05, 0.10, 0.15, 0.20, and 0.25 %) were prepared to a final volume of 200 mL using deionized water; 20 larvae of each instars (I-IV) and pupa were exposed to each test concentration separately which included a set of control (distilled water) group with five replicates. Characterization of the synthesized Ag NPs were about 10-20 nm without aggregation. Susceptibility of larval instars to synthesized Ag NPs was higher than the extracellular filtrate of T. harzianum alone after 24-h exposure, where the highest mortality was recorded as 92 and 96 % for first and second instars and

Impacts of biocontrol products on Rhizoctonia disease of potato and soil microbial communities, and their persistence in soil

USDA-ARS?s Scientific Manuscript database

Four commercial biocontrol formulations (Bacillus subtilis GB03, Burkholderia ambifaria type Wisconsin isolate J82, Trichoderma virens Gl-21, and Trichoderma harzianum strain T-22), a chemical seed treatment (Topsin, mancozeb, and cymoxanil mixture, TMC), and a combination chemical/biological treatm...

Combination of Trichoderma harzianum endochitinase and a membrane-affecting fungicide on control of Alternaria leaf spot in transgenic broccoli plants.

PubMed

Mora, A; Earle, E D

2001-04-01

Progeny from transgenic broccoli (cv. Green Comet) expressing a Trichoderma harzianum endochitinase gene were used to assess the interaction between endochitinase and the fungicide Bayleton in the control of Alternaria brassicicola. In vitro assays have shown synergistic effects of endochitinase and fungicides on fungal pathogens. Our study examined the in planta effects of endochitinase and Bayleton, individually and in combination. Two month old transgenic and non-transgenic plants were sprayed with ED50 levels of Bayleton and/or inoculated with an A. brassicicola spore suspension. Disease levels in non-sprayed transgenic plants were not statistically different from sprayed transgenic plants nor from sprayed non-transgenic controls. Thus endochitinase-transgenic plants alone provided a significant reduction of disease severity, comparable to the protection by fungicide on non-transgenic plants. Comparison of the expected additive and observed effects revealed no synergism between endochitinase and Bayleton (at ED50 level), and usually less than an additive effect. Some transgenic lines sprayed with fungicide at doses higher than ED50 showed resistance similar to the non-sprayed transgenic lines, again suggesting no synergistic effect. Lack of synergism may be due to incomplete digestion of the cell wall by endochitinase, so that the effect of Bayleton at the cell membrane is not enhanced.

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants.

PubMed

Janoušková, Martina; Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation.

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants

PubMed Central

Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation. PMID:28738069

Evaluation of Series T22 Wet Tantalum Capacitors

NASA Technical Reports Server (NTRS)

Teverovsky, Alexander

2017-01-01

Several types of advanced wet tantalum capacitors, and series T22 in particular, are designed without internal Teflon sealing that is used for military grade, CLR style capacitors. This raises concerns regarding hermeticity of the single seal parts and their capability to withstand high internal gas pressures that might develop during operation in space. To address these issues, T22 series capacitors rated to 50 V and 125 V were subjected to highly accelerated life testing (HALT) at 125 C and rated voltage and step stress random vibration testing (RVT). To simulate conditions of storage or operation under increased internal gas pressure, the parts were stored at temperature of 150 C for 2500 hr (HTS150). Electrical characteristics of the parts were measured through the storage testing and the hermeticity leak rate was tested before and after HTS150. To assess thermo-mechanical robustness of the part, capacitors were manually soldered onto printed wired boards (PWB) and stressed by 1000 temperature cycles between -55 C and +125 C. The effect of temperature cycling was assessed by additional HALT at different temperatures. Results show that T22 series capacitors have robust design and can satisfy requirements for space applications.

Renal cell carcinoma and a constitutional t(11;22)(q23;q11.2): case report and review of the potential link between the constitutional t(11;22) and cancer.

PubMed

Doyen, Jérôme; Carpentier, Xavier; Haudebourg, Juliette; Hoch, Benjamin; Karmous-Benailly, Houda; Ambrosetti, Damien; Fabas, Thibault; Amiel, Jean; Lambert, Jean-Claude; Pedeutour, Florence

2012-11-01

We observed a t(11;22)(q23-24;q11.2-12) and monosomy 3 in renal tumor cells from a 72-year-old man. The hypothesis of a primitive peripheral neuroectodermal tumor (PPNET) located in the kidney was promptly excluded: Histologically, the tumor was a clear cell renal cell carcinoma (RCC) and we did not observe an EWSR1 gene rearrangement. The constitutional origin of this alteration was established. We report on the second case of RCC in a patient with a constitutional t(11;22). The t(11;22)(q23;q11.2) is the main recurrent germline translocation in humans. Unbalanced translocation can be transmitted to the progeny and can cause Emanuel syndrome. Our observation alerts cancer cytogeneticists to the fortuitous discovery of the constitutional t(11;22) in tumor cells. This translocation appears grossly similar to the t(11;22)(q24;q12) of PPNET and should be evoked if present in all cells of a tumor other than PPNET. This is important when providing appropriate genetic counseling. Moreover, the potential oncogenic role of the t(11;22) and its predisposing risk of cancer are under debate. The family history of the patient revealed a disabled brother who died at an early age from colon cancer and a sister with breast cancer. This observation reopens the issue of a link between the constitutional t(11;22) and cancer, and the utility of cancer prevention workups for t(11;22) carriers. Copyright © 2012 Elsevier Inc. All rights reserved.

Effective production of biologically active water-soluble β-1,3-glucan by a coupled system of Agrobacterium sp. and Trichoderma harzianum.

PubMed

Liang, Ying; Zhu, Li; Gao, Minjie; Wu, Jianrong; Zhan, Xiaobei

2018-05-28

Water-soluble β-1,3-glucan (w-glucan) prepared from curdlan is reported to possess various bioactive and medicinal properties. To develop an efficient and cost-effective microbial fermentation method for the direct production of w-glucan, a coupled fermentation system of Agrobacterium sp. and Trichoderma harzianum (CFS-AT) was established. The effects of Tween-80, glucose flow rate, and the use of a dissolved oxygen (DO) control strategy on w-glucan production were assessed. The addition of 10 g L -1 Tween-80 to the CFS-AT enhanced w-glucan production, presumably by loosening the curdlan ultrastructure and increasing the efficiency of curdlan hydrolysis. A two-stage glucose and DO control strategy was optimal for w-glucan production. At the T. harzianum cell growth stage, the optimal glucose flow rate and agitation speed were 2.0 g L -1 hr -1 and 600 rpm, respectively, and at the w-glucan production stage, they were 0.5 g L -1 hr -1 and 400 rpm, respectively. W-glucan production reached 17.31 g L -1 , with a degree of polymerization of 19-25. Furthermore, w-glucan at high concentrations exhibited anti-tumor activity against MCF-7, HepG2, and Hela cancer cells in vitro. This study provides a novel, cost-effective, eco-friendly, and efficient microbial fermentation method for the direct production of biologically active w-glucan.

Differential response of kabuli and desi chickpea genotypes toward inoculation with PGPR in different soils

PubMed Central

Imran, Asma; Mirza, Muhammad S.; Shah, Tariq M.; Malik, Kauser A.; Hafeez, Fauzia Y.

2015-01-01

Pakistan is among top three chickpea producing countries but the crop is usually grown on marginal lands without irrigation and fertilizer application which significantly hampers its yield. Soil fertility and inoculation with beneficial rhizobacteria play a key role in nodulation and yield of legumes. Four kabuli and six desi chickpea genotypes were, therefore, evaluated for inoculation response with IAA-producing Ochrobactrum ciceri Ca-34T and nitrogen fixing Mesorhizobium ciceri TAL-1148 in single and co-inoculation in two soils. The soil type 1 was previously unplanted marginal soil having low organic matter, P and N contents compared to soil type 2 which was a fertile routinely legume-cultivated soil. The effect of soil fertility status was pronounced and fertile soil on average, produced 31% more nodules, 62% more biomass and 111% grain yield than marginal soil. Inoculation either with O. ciceri alone or its co-inoculation with M. ciceri produced on average higher nodules (42%), biomass (31%), grains yield (64%) and harvest index (72%) in both chickpea genotypes over non-inoculated controls in both soils. Soil 1 showed maximum relative effectiveness of Ca-34T inoculation for kabuli genotypes while soil 2 showed for desi genotypes except B8/02. Desi genotype B8/02 in soil type 1 and Pb-2008 in soil type 2 showed significant yield increase as compared to respective un-inoculated controls. Across bacterial inoculation treatments, grain yield was positively correlated to growth and yield contributing parameters (r = 0.294* to 0.838*** for desi and r = 0.388* to 0.857** for kabuli). PCA and CAT-PCA analyses clearly showed a site-specific response of genotype x bacterial inoculation. Furthermore, the inoculated bacterial strains were able to persist in the rhizosphere showing colonization on root and within nodules. Present study shows that plant growth promoting rhizobacteria (PGPR) inoculation should be integrated with national chickpea breading program in

Phosphate dissolving fungi: Mechanism and application in alleviation of salt stress in wheat.

PubMed

Gaind, Sunita

2016-12-01

The present investigation reveals the solubilization efficiency of tri-calcium phosphate (TCP), Udaipur rock phosphate (URP), aluminium phosphate (AP) and ferric phosphate (FP) by Aspergillus niger (ITCC 6719) and Trichoderma harzianum (ITCC 6721) as function of carbon concentrations. Increasing glucose concentration from 1 to 7% in the growth medium, though improved the phosphorus (P) solubilization significantly but each fungal strain preferred different optimum carbon concentrations for mediating solubilization of different P sources. The two fungi employed different mechanisms to reduce medium pH for release of P from TCP, AP and FP. However, URP was solubilized solely through fungal production of citric, succinic, propionic, malic and acetic acid. A linear increase in citric acid production with increasing carbon concentration was recorded during FP solubilization by T. harzianum. The cell free culture filtrate of A. niger detected high phytase and low acid phosphatase activity titre whereas results were vice versa for T. harzianum. Both the fungal strains possessed plant growth promoting attributes such as auxin and sidreophore production and could solubilize Zn. In hydroponic system (with 60mM of sodium chloride concentration), supplementation with culture filtrate from each fungal strain increased the shoot growth of wheat seedlings significantly compared to non culture filtrate control. Use of A.niger as bio-inoculant could be a sustainable approach to improve soil P availability, promote plant growth and alleviate adverse effect of salt stress. Copyright © 2016 Elsevier GmbH. All rights reserved.

Observations on root disease of container whitebark pine seedlings treated with biological controls

Treesearch

R. Kasten Dumroese

2008-01-01

I observed that whitebark pine (Pinus albicaulis Engelm. [Pinaceae]) germinants treated with biological controls, one commercially available (Trichoderma harzianum strain T-22), and the other being studied for potential efficacy (Fusarium oxysporum isolate Q12), experienced less seedling mortality caused by root disease than did a...

Maturation of green waste compost as affected by inoculation with the white-rot fungi Trametes versicolor and Phanerochaete chrysosporium.

PubMed

Gong, Xiaoqiang; Li, Suyan; Sun, Xiangyang; Zhang, Lu; Zhang, Tao; Wei, Le

2017-04-01

Green waste was separately inoculated on day 0 and day 14 with either Trametes versicolor or Phanerochaete chrysosporium to determine their effects on composting time and compost quality. Inoculation with T. versicolor and P. chrysosporium caused more rapid and higher increases in compost temperatures, increased the duration of the thermophilic temperature stage, and reduced the maturity time. Inoculation with T. versicolor and P. chrysosporium greatly increased the quality of the final composts in terms of pH, electrical conductivity, organic matter concentration, C/N ratio, germination index, and nutrient content. Inoculation with T. versicolor and P. chrysosporium also significantly increased the degradation of lignin by 7.1% and 8.2%, respectively, and increased the degradation of cellulose by 10.6% and 13.6%, respectively.

Solid state production of polygalacturonase and xylanase by Trichoderma species using cantaloupe and watermelon rinds.

PubMed

Mohamed, Saleh A; Al-Malki, Abdulrahman L; Khan, Jalaluddin A; Kabli, Saleh A; Al-Garni, Saleh M

2013-10-01

Different solid state fermentation (SSF) sources were tested such as cantaloupe and watermelon rinds, orange and banana peels, for the production of polygalacturonase (PG) and xylanase (Xyl) by Trichoderma harzianum and Trichoderma virens. The maximum production of both PG and Xyl were obtained by T. harzianum and T. virnes grown on cantaloupe and watermelon rinds, respectively. Time course, moisture content, temperature, pH, supplementation with carbon and nitrogen sources were optimized to achieve the maximum production of both PG and Xyl of T. harzianum and T. virens using cantaloupe and watermelon rinds, respectively. The maximum production of PG and Xyl of T. harzianum and T. virens was recorded at 4-5 days of incubation, 50-66% moisture, temperature 28-35°C and pH 6-7. The influence of supplementary carbon and nitrogen sources was studied. For T. harzianum, lactose enhanced PG activity from 87 to 120 units/g solid, where starch and maltose enhanced Xyl activity from 40 to 55-60 units/g solid for T. virnes. Among the nitrogen sources, ammonium sulphate, ammonium nitrate, yeast extract and urea increased PG activity from 90 to 110-113 units/g solid for T. harzianum. Similarly, ammonium chloride, ammonium sulphate and yeast extract increased Xyl activity from 45 to 55-70 units/g solid for T. virens.

Fungal Screening on Olive Oil for Extracellular Triacylglycerol Lipases: Selection of a Trichoderma harzianum Strain and Genome Wide Search for the Genes

PubMed Central

Canseco-Pérez, Miguel Angel; Castillo-Avila, Genny Margarita; Islas-Flores, Ignacio; Apolinar-Hernández, Max M.; Rivera-Muñoz, Gerardo; Gamboa-Angulo, Marcela; Couoh-Uicab, Yeny

2018-01-01

A lipolytic screening with fungal strains isolated from lignocellulosic waste collected in banana plantation dumps was carried out. A Trichoderma harzianum strain (B13-1) showed good extracellular lipolytic activity (205 UmL−1). Subsequently, functional screening of the lipolytic activity on Rhodamine B enriched with olive oil as the only carbon source was performed. The successful growth of the strain allows us to suggest that a true lipase is responsible for the lipolytic activity in the B13-1 strain. In order to identify the gene(s) encoding the protein responsible for the lipolytic activity, in silico identification and characterization of triacylglycerol lipases from T. harzianum is reported for the first time. A survey in the genome of this fungus retrieved 50 lipases; however, bioinformatic analyses and putative functional descriptions in different databases allowed us to choose seven lipases as candidates. Suitability of the bioinformatic screening to select the candidates was confirmed by reverse transcription polymerase chain reaction (RT-PCR). The gene codifying 526309 was expressed when the fungus grew in a medium with olive oil as carbon source. This protein shares homology with commercial lipases, making it a candidate for further applications. The success in identifying a lipase gene inducible with olive oil and the suitability of the functional screening and bioinformatic survey carried out herein, support the premise that the strategy can be used in other microorganisms with sequenced genomes to search for true lipases, or other enzymes belonging to large protein families. PMID:29370083

Biological Control of Alternaria Fruit Rot of Chili by Trichoderma Species under Field Conditions

PubMed Central

Rahman, M. A.; Alam, M. Firoz

2010-01-01

Trichoderma strains were evaluated under field conditions to assay their efficacy in suppressing Alternaria fruit rot disease and promoting chili plant growth. The experiment was conducted at the Botanical Garden, Rajshahi University, Bangladesh from July 2006 to March 2007. Application of Trichoderma harzianum IMI 392432 significantly (p = 0.05) suppressed the disease compared to Alternaria tenuis (T2) treatment and improved both growth and yield. The treatment T4 (T. harzianum IMI-392432 + A. tenuis) was most effective in reducing disease percentage (72.27%) compared to A. tenuis (T1) treatment. The highest seed germination rate (85.56%) and the highest growth and yield (12.5 g/plant) was also recorded in the same treatment (T4), followed by T5 (T. harzianum IMI-392433 000000 + A. tenuis), T6 (T. harzianum IMI-392434 +A. tenuis), T2 (T. virens IMI-392430 + A. tenuis), and T3 (T. pseudokoningii IMI-392431 +A. tenuis) treatment, while single treatment with A. tenuis significantly decreased these values. PMID:23956637

T Cell Inactivation by Poxviral B22 Family Proteins Increases Viral Virulence

PubMed Central

Alzhanova, Dina; Hammarlund, Erika; Reed, Jason; Meermeier, Erin; Rawlings, Stephanie; Ray, Caroline A.; Edwards, David M.; Bimber, Ben; Legasse, Alfred; Planer, Shannon; Sprague, Jerald; Axthelm, Michael K.; Pickup, David J.; Lewinsohn, David M.; Gold, Marielle C.; Wong, Scott W.; Sacha, Jonah B.; Slifka, Mark K.; Früh, Klaus

2014-01-01

Infections with monkeypox, cowpox and weaponized variola virus remain a threat to the increasingly unvaccinated human population, but little is known about their mechanisms of virulence and immune evasion. We now demonstrate that B22 proteins, encoded by the largest genes of these viruses, render human T cells unresponsive to stimulation of the T cell receptor by MHC-dependent antigen presentation or by MHC-independent stimulation. In contrast, stimuli that bypass TCR-signaling are not inhibited. In a non-human primate model of monkeypox, virus lacking the B22R homologue (MPXVΔ197) caused only mild disease with lower viremia and cutaneous pox lesions compared to wild type MPXV which caused high viremia, morbidity and mortality. Since MPXVΔ197-infected animals displayed accelerated T cell responses and less T cell dysregulation than MPXV US2003, we conclude that B22 family proteins cause viral virulence by suppressing T cell control of viral dissemination. PMID:24832205

T cell inactivation by poxviral B22 family proteins increases viral virulence.

PubMed

Alzhanova, Dina; Hammarlund, Erika; Reed, Jason; Meermeier, Erin; Rawlings, Stephanie; Ray, Caroline A; Edwards, David M; Bimber, Ben; Legasse, Alfred; Planer, Shannon; Sprague, Jerald; Axthelm, Michael K; Pickup, David J; Lewinsohn, David M; Gold, Marielle C; Wong, Scott W; Sacha, Jonah B; Slifka, Mark K; Früh, Klaus

2014-05-01

Infections with monkeypox, cowpox and weaponized variola virus remain a threat to the increasingly unvaccinated human population, but little is known about their mechanisms of virulence and immune evasion. We now demonstrate that B22 proteins, encoded by the largest genes of these viruses, render human T cells unresponsive to stimulation of the T cell receptor by MHC-dependent antigen presentation or by MHC-independent stimulation. In contrast, stimuli that bypass TCR-signaling are not inhibited. In a non-human primate model of monkeypox, virus lacking the B22R homologue (MPXVΔ197) caused only mild disease with lower viremia and cutaneous pox lesions compared to wild type MPXV which caused high viremia, morbidity and mortality. Since MPXVΔ197-infected animals displayed accelerated T cell responses and less T cell dysregulation than MPXV US2003, we conclude that B22 family proteins cause viral virulence by suppressing T cell control of viral dissemination.

Comparison of Inoculation with the InoqulA and WASP Automated Systems with Manual Inoculation

PubMed Central

Croxatto, Antony; Dijkstra, Klaas; Prod'hom, Guy

2015-01-01

The quality of sample inoculation is critical for achieving an optimal yield of discrete colonies in both monomicrobial and polymicrobial samples to perform identification and antibiotic susceptibility testing. Consequently, we compared the performance between the InoqulA (BD Kiestra), the WASP (Copan), and manual inoculation methods. Defined mono- and polymicrobial samples of 4 bacterial species and cloudy urine specimens were inoculated on chromogenic agar by the InoqulA, the WASP, and manual methods. Images taken with ImagA (BD Kiestra) were analyzed with the VisionLab version 3.43 image analysis software to assess the quality of growth and to prevent subjective interpretation of the data. A 3- to 10-fold higher yield of discrete colonies was observed following automated inoculation with both the InoqulA and WASP systems than that with manual inoculation. The difference in performance between automated and manual inoculation was mainly observed at concentrations of >106 bacteria/ml. Inoculation with the InoqulA system allowed us to obtain significantly more discrete colonies than the WASP system at concentrations of >107 bacteria/ml. However, the level of difference observed was bacterial species dependent. Discrete colonies of bacteria present in 100- to 1,000-fold lower concentrations than the most concentrated populations in defined polymicrobial samples were not reproducibly recovered, even with the automated systems. The analysis of cloudy urine specimens showed that InoqulA inoculation provided a statistically significantly higher number of discrete colonies than that with WASP and manual inoculation. Consequently, the automated InoqulA inoculation greatly decreased the requirement for bacterial subculture and thus resulted in a significant reduction in the time to results, laboratory workload, and laboratory costs. PMID:25972424

Toxoplasma gondii: infection natural congenital in cattle and an experimental inoculation of gestating cows with oocysts.

PubMed

Costa, Gustavo Henrique Nogueira; da Costa, Alvimar José; Lopes, Welber Daniel Zanetti; Bresciani, Katia Denise Saraiva; dos Santos, Thais Rabelo; Esper, César Roberto; Santana, Aureo Evangelista

2011-01-01

Two studies, of a natural infection and an experimental infection, were performed in order to study congenital transmission of Toxoplasma gondii in cattle. In the first study, 50 fetuses were harvested from gestating cows that were eutanasied at a municipal slaughterhouse in Jaboticabal, São Paulo state, Brazil. In the second study, 11 gestating cows were divided into four groups for inoculation with T. gondii: GI consisted of three cows inoculated with 1.0 × 10(5) oocysts during their first trimester of gestation; GII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their second trimester of gestation; GIII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their last trimester of gestation; and GIV consisted of two control cows, one during its first and the other during its second trimester of gestation. In both studies, the presence of T. gondii was confirmed both indirectly by immunofluorescence assay (IFAT). In the natural infection experiment, 18% (9/50) of the gestating cows were confirmed to have specific antibodies (IFAT--1:64) against T. gondii. The bioassay was able to diagnose the presence of T. gondii in the tissue samples from three calves. In the second experiment, the nine cows from groups I, II and III presented with specific antibodies (IFAT) against T. gondii. In contrast, T. gondii could not be detected by IFAT, histopathological examination or the bioassay in any of the nine calves born to cows experimentally infected with T. gondii oocysts. Based on the results from both studies, we conclude that congenital infection of T. gondii in cattle, while infrequent, does occur naturally. The pathogenicity of the strain of T. gondii may influence the likelihood of this route of transmission. Copyright © 2010 Elsevier Inc. All rights reserved.

Co-composting of spent coffee ground with olive mill wastewater sludge and poultry manure and effect of Trametes versicolor inoculation on the compost maturity.

PubMed

Hachicha, Ridha; Rekik, Olfa; Hachicha, Salma; Ferchichi, Mounir; Woodward, Steve; Moncef, Nasri; Cegarra, Juan; Mechichi, Tahar

2012-07-01

The co-composting of spent coffee grounds, olive mill wastewater sludge and poultry manure was investigated on a semi-industrial scale. In order to reduce the toxicity of the phenolic fraction and to improve the degree of composting humification, composts were inoculated with the white-rot fungus Trametes versicolor in the early stages of the maturation phase. During composting, a range of physico-chemical parameters (temperature and both organic matter and C/N reduction), total organic carbon, total nitrogen, elemental composition, lignin degradation and spectroscopic characteristics of the humic acids (HAs) were determined; impacts of the composting process on germination index of Hordeum vulgare and Lactuca sativa were assessed. The coffee waste proved to be a highly compostable feedstock, resulting in mature final compost with a germination index of 120% in less than 5 months composting. In addition, inoculation with T. versicolor led to a greater degree of aromatization of HA than in the control pile. Moreover, in the inoculated mixture, lignin degradation was three times greater and HA increased by 30% (P<0.05), compared to the control pile. In the T. versicolor inoculated mixture, the averages of C and N were significantly enhanced in the HA molecules (P<0.05), by 26% and 22%, respectively. This improvement in the degree of humification was confirmed by the ratio of optical densities of HA solutions at 465 and 665 nm which was lower for HA from the treated mixture (4.5) than that from the control pile (5.4). Copyright © 2012 Elsevier Ltd. All rights reserved.

Correlation of structure, function and protein dynamics in GH7 cellobiohydrolases from Trichoderma atroviride, T. reesei and T. harzianum.

PubMed

Borisova, Anna S; Eneyskaya, Elena V; Jana, Suvamay; Badino, Silke F; Kari, Jeppe; Amore, Antonella; Karlsson, Magnus; Hansson, Henrik; Sandgren, Mats; Himmel, Michael E; Westh, Peter; Payne, Christina M; Kulminskaya, Anna A; Ståhlberg, Jerry

2018-01-01

The ascomycete fungus Trichoderma reesei is the predominant source of enzymes for industrial conversion of lignocellulose. Its glycoside hydrolase family 7 cellobiohydrolase (GH7 CBH) Tre Cel7A constitutes nearly half of the enzyme cocktail by weight and is the major workhorse in the cellulose hydrolysis process. The orthologs from Trichoderma atroviride ( Tat Cel7A) and Trichoderma harzianum ( Tha Cel7A) show high sequence identity with Tre Cel7A, ~ 80%, and represent naturally evolved combinations of cellulose-binding tunnel-enclosing loop motifs, which have been suggested to influence intrinsic cellobiohydrolase properties, such as endo-initiation, processivity, and off-rate. The Tat Cel7A, Tha Cel7A, and Tre Cel7A enzymes were characterized for comparison of function. The catalytic domain of Tat Cel7A was crystallized, and two structures were determined: without ligand and with thio-cellotriose in the active site. Initial hydrolysis of bacterial cellulose was faster with Tat Cel7A than either Tha Cel7A or Tre Cel7A. In synergistic saccharification of pretreated corn stover, both Tat Cel7A and Tha Cel7A were more efficient than Tre Cel7A, although Tat Cel7A was more sensitive to thermal inactivation. Structural analyses and molecular dynamics (MD) simulations were performed to elucidate important structure/function correlations. Moreover, reverse conservation analysis (RCA) of sequence diversity revealed divergent regions of interest located outside the cellulose-binding tunnel of Trichoderma spp. GH7 CBHs. We hypothesize that the combination of loop motifs is the main determinant for the observed differences in Cel7A activity on cellulosic substrates. Fine-tuning of the loop flexibility appears to be an important evolutionary target in Trichoderma spp., a conclusion supported by the RCA data. Our results indicate that, for industrial use, it would be beneficial to combine loop motifs from Tat Cel7A with the thermostability features of Tre Cel7A. Furthermore

Inoculating for smallpox.

PubMed

Greene, Jan

2003-04-01

In California, one hospital decided the time was right to inoculate staff volunteers with the smallpox vaccine. Weighing the same pros and cons--civic responsibility, health risk, cost and reluctance of staff--other hospitals opted out of inoculations, at least for now. What led these organizations to such divergent decisions?

Swine dysentery: inoculation of gnotobiotic pigs with Treponema hyodysenteriae and Vibrio coli and a Peptostreptococcus.

PubMed Central

Brandenburg, A C; Miniats, O P; Geissinger, H D; Ewert, E

1977-01-01

Pure cultures of Treponema hyodysenteriae given orally to conventional pigs resulted in the development of swine dysentery, whereas identical cultures given to gnotobiotic pigs did not produce the disease. Oral inoculation of gnotobiotic pigs with Vibrio coli and/or a peptostreptococcus in addition to T. hyodysenteriae did not result in dysentery. Neutralization of gastric secretions with NaHCO3 immediately prior to inoculation with T. hyodysenteriae increased the period during which treponemes were evident in the feces, as did the inoculation of this organism via the intracecal route. None of the gnotobiotic pigs with a persistent fecal Treponema population developed signs of dysentery. Factors other than those investigated in this work must play a part in the etiology of swine dysentery. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:907906

Potential of Trichoderma harzianum for control of banana leaf fungal pathogens when applied with a food source and an organic adjuvant.

PubMed

Samuelian, Suren

2016-06-01

Trichoderma isolates were obtained from diseased leaves and fruit collected from plantations in the main banana production area in Northern Queensland. Phylogenetic analyses identified the Trichoderma isolates as T. harzianum and T. virens. The Trichoderma spp. were found to be antagonistic against the banana leaf pathogens Mycosphaerella musicola, Cordana musae, and Deightoniella torulosa in vitro. Several products used by the banana industry to increase production, including molasses, Fishoil and Seasol, were tested as food source for the Trichoderma isolates. The optimal food substrate was found to be molasses at a concentration of 5 %, which when used in combination with a di-1-p-menthene spreader-sticker enhanced the survivability of Trichoderma populations under natural conditions. This formulation suppressed D. torulosa development under glasshouse conditions. Furthermore, high sensitivity was observed towards the protectant fungicide Mancozeb but Biopest oil ® , a paraffinic oil, only marginally suppressed the growth of Trichoderma isolates in vitro. Thus, this protocol represents a potential to manage banana leaf pathogens as a part of an integrated disease approach.

Antitumor HPV E7-specific CTL activity elicited by in vivo engineered exosomes produced through DNA inoculation.

PubMed

Di Bonito, Paola; Chiozzini, Chiara; Arenaccio, Claudia; Anticoli, Simona; Manfredi, Francesco; Olivetta, Eleonora; Ferrantelli, Flavia; Falcone, Emiliana; Ruggieri, Anna; Federico, Maurizio

2017-01-01

We recently proved that exosomes engineered in vitro to deliver high amounts of HPV E7 upon fusion with the Nef mut exosome-anchoring protein elicit an efficient anti-E7 cytotoxic T lymphocyte immune response. However, in view of a potential clinic application of this finding, our exosome-based immunization strategy was faced with possible technical difficulties including industrial manufacturing, cost of production, and storage. To overcome these hurdles, we designed an as yet unproven exosome-based immunization strategy relying on delivery by intramuscular inoculation of a DNA vector expressing Nef mut fused with HPV E7. In this way, we predicted that the expression of the Nef mut /E7 vector in muscle cells would result in a continuous source of endogenous (ie, produced by the inoculated host) engineered exosomes able to induce an E7-specific immune response. To assess this hypothesis, we first demonstrated that the injection of a Nef mut /green fluorescent protein-expressing vector led to the release of fluorescent exosomes, as detected in plasma of inoculated mice. Then, we observed that mice inoculated intramuscularly with a vector expressing Nef mut /E7 developed a CD8 + T-cell immune response against both Nef and E7. Conversely, no CD8 + T-cell responses were detected upon injection of vectors expressing either the wild-type Nef isoform of E7 alone, most likely a consequence of their inefficient exosome incorporation. The production of immunogenic exosomes in the DNA-injected mice was formally demonstrated by the E7-specific CD8 + T-cell immune response we detected in mice inoculated with exosomes isolated from plasma of mice inoculated with the Nef mut /E7 vector. Finally, we provide evidence that the injection of Nef mut /E7 DNA led to the generation of effective antigen-specific cytotoxic T lymphocytes whose activity was likely part of the potent, therapeutic antitumor effect we observed in mice implanted with TC-1 tumor cells. In summary, we established

Mitigation of salt stress in white clover (Trifolium repens) by Azospirillum brasilense and its inoculation effect.

PubMed

Khalid, Muhammad; Bilal, Muhammad; Hassani, Danial; Iqbal, Hafiz M N; Wang, Hang; Huang, Danfeng

2017-12-01

Salinity is one of the increasingly serious environmental problems worldwide for cultivating agricultural crops. The present study was aimed to ascertain the potential of beneficial soil bacterium Azospirillum brasilense to alleviate saline stress in Trifolium repens. Experimental plants (white clover) were grown from seeds and inoculated with or without A. brasilense bacterial strain supplemented with 0, 40, 80, or 120 mM NaCl into soil. The growth attributes including, shoot heights, root lengths, fresh and dry weights, leaf area and chlorophyll content were significantly enhanced in T. repens plants grown in A. brasilense inoculated soil than un-inoculated controls, particularly under elevated salinity conditions (40, 80 and 120 mM NaCl). Malondialdehyde content of leaf was recorded to be declined under saline conditions. Moreover, the K + /Na + ratio was also improved in bacterium-inoculated plants, since A. brasilense significantly reduced the root and shoot Na + level under high salty environment. Results revealed that soil inoculation with A. brasilense could significantly promote T. repens growth under both non-saline and saline environments, and this study might be extended to other vegetables and crops for the germination and growth enhancement.

Media optimization for laccase production by Trichoderma harzianum ZF-2 using response surface methodology.

PubMed

Gao, Huiju; Chu, Xiang; Wang, Yanwen; Zhou, Fei; Zhao, Kai; Mu, Zhimei; Liu, Qingxin

2013-12-01

Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from Shandong, China, and showed dye decolorization activities. The objective of this study was to optimize its culture conditions using a statistical analysis of its laccase production. The interactions between different fermentation parameters for laccase production were characterized using a Plackett-Burman design and the response surface methodology. The different media components were initially optimized using the conventional one-factor-at-a-time method and an orthogonal test design, and a Plackett-Burman experiment was then performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal, and CuSO4 were all found to have a significant influence on laccase production, and the optimal concentrations of these three factors were then sequentially investigated using the response surface methodology with a central composite design. The resulting optimal medium components for laccase production were determined as follows: wheat straw powder 7.63 g/l, soybean meal 23.07 g/l, (NH4)2SO4 1 g/l, CuSO4 0.51 g/l, Tween-20 1 g/l, MgSO4 1 g/l, and KH2PO4 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased 59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized medium. This is the first report on the statistical optimization of laccase production by Trichoderma harzianum ZF-2.

Effects of letrozole on breast cancer micro-metastatic tumor growth in bone and lung in mice inoculated with murine 4T1 cells.

PubMed

Wang, Wendan; Belosay, Aashvini; Yang, Xujuan; Hartman, James A; Song, Huaxin; Iwaniec, Urszula T; Turner, Russell T; Churchwell, Mona I; Doerge, Daniel R; Helferich, William G

2016-06-01

Breast cancer (BC) is the leading cancer in women worldwide. Metastasis occurs in stage IV BC with bone and lung being common metastatic sites. Here we evaluate the effects of the aromatase inhibitor letrozole on BC micro-metastatic tumor growth in bone and lung metastasis in intact and ovariectomized (OVX) mice with murine estrogen receptor negative (ER-) BC cells inoculated in tibia. Forty-eight BALB/c mice were randomly assigned to one of four groups: OVX, OVX + Letrozole, Intact, and Intact + Letrozole, and injected with 4T1 cells intra-tibially. Letrozole was subcutaneously injected daily for 23 days at a dose of 1.75 µg/g body weight. Tumor progression was monitored by bioluminescence imaging (BLI). Following necropsy, inoculated tibiae were scanned via µCT and bone response to tumor was scored from 0 (no ectopic mineralization/osteolysis) to 5 (extensive ectopic mineralization/osteolysis). OVX mice had higher tibial pathology scores indicative of more extensive bone destruction than intact mice, irrespective of letrozole treatment. Letrozole decreased serum estradiol levels and reduced lung surface tumor numbers in intact animals. Furthermore, mice receiving letrozole had significantly fewer tumor colonies and fewer proliferative cells in the lung than OVX and intact controls based on H&E and Ki-67 staining, respectively. In conclusion, BC-inoculated OVX animals had higher tibia pathology scores than BC-inoculated intact animals and letrozole reduced BC metastases to lungs. These findings suggest that, by lowering systemic estrogen level and/or by interacting with the host organ, the aromatase inhibitor letrozole has the potential to reduce ER- BC metastasis to lung.

γδ T cell receptors recognize the non-classical major histocompatibility complex (MHC) molecule T22 via conserved anchor residues in a MHC peptide-like fashion.

PubMed

Sandstrom, Andrew; Scharf, Louise; McRae, Gabrielle; Hawk, Andrew J; Meredith, Stephen C; Adams, Erin J

2012-02-17

The molecular mechanisms by which γδ T cells recognize ligand remain a mystery. The non-classical MHC molecule T22 represents the best characterized ligand for murine γδ T cells, with a motif (W … EGYEL) present in the γδ T cell receptor complementary-determining region 3δ (CDR3δ) loop mediating γδ T cell recognition of this molecule. Produced through V(D)J recombination, this loop is quite diverse, with different numbers and chemical types of amino acids between Trp and EGYEL, which have unknown functional consequences for T22 recognition. We have investigated the biophysical and structural effects of CDR3δ loop diversity, revealing a range of affinities for T22 but a common thermodynamic pattern. Mutagenesis of these CDR3δ loops defines the key anchor residues involved in T22 recognition as W … EGYEL, similar to those found for the G8 CDR3δ loop, and demonstrates that spacer residues modulate but are not required for T22 recognition. Comparison of the location of these residues in the T22 interface reveals a striking similarity to peptide anchor residues in classically presented MHC peptides, with the key Trp residue of the CDR3δ motif completing the deficient peptide-binding groove of T22. This suggests that γδ T cell recognition of T22 utilizes the conserved ligand-presenting nature of the MHC fold.

Identification of melatonin in Trichoderma spp. and detection of melatonin content under controlled-stress growth conditions from T. asperellum.

PubMed

Liu, Tong; Zhao, Fengzhou; Liu, Zhen; Zuo, Yuhu; Hou, Jumei; Wang, Yanjie

2016-07-01

T. koningii, T. harzianum, T. asperellum, T. longibrachiatum, and T. viride were analyzed using liquid chromatography-tandem mass spectrometry to determine whether melatonin is present. Results showed that there were abundant amounts of endogenous melatonin in five Trichoderma species, but no melatonin was found in any of the culture filtrates. T. asperellum had the highest amount of melatonin (27.588 ± 0.326 μg g(-1) dry mass), followed by T. koningii, T. harzianum, T. longibrachiatum, and T. viride. The endogenous melatonin content of T. asperellum in controlled-stress growth conditions was also detected. The data showed that chemical stressors (CdCl2 , CuSO4 , and H2 O2 ) provoked an increase in endogenous melatonin levels. CdCl2 had the highest stimulatory effect on melatonin production, as the product reached reaching up to three times the melatonin content of the control. NaCl stimulated a decrease of melatonin. Acidic conditions (pH 3 and pH 5) as well as slightly alkaline conditions (pH 9) resulted in an increase in the melatonin content, whereas pH11 resulted in a significant decrease in the melatonin content, only 12.276 ± 0.205 μg g(-1) dry mass. The current study is first to report melatonin content and the change of melatonin content under different stress situations in Trichoderma spp. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Combined Effects of Inoculation Level and Sequence on Biochemical and Microbiological Characteristics of Probiotic Doogh

PubMed Central

Rahmati Roudsari, Mohammad; Sohrabvandi, Sara; Homayouni Rad, Aziz; Mortazavian, Amir Mohammad

2013-01-01

The combined effects of inoculation level (4 or 8-fold compared to standard inoculation) and sequence (standard inoculation before fermentation and 3-fold inoculation at the end of fermentation=1+3, Two-fold inoculation before fermentation and the same at the end of fermentation=2+2, 2+6, 4-fold before fermentation=4, 4+4, and 8) of culture inoculum containing probiotics on biochemical and microbiological characteristics of probiotic Doogh during fermentation and over 21 days of refrigerated storage (4˚C) were investigated. The probiotic microorganisms were L. acidophilus LA-5 and Bifidobacterium lactis BB-12. Overall, the treatments 8, 4 and 4+4 resulted in the highest viability at the end of fermentation as well as at early days of refrigerated storage. During the second half of cold storage period, the greatest viability of probiotics was related to the treatment 2+6. The treatment ‘8’ showed the shortest incubation time as well as the highest pH drop rate and acidity increase rate during fermentation and over the storage period. PMID:24250636

17 CFR 240.11a2-2(T) - Transactions effected by exchange members through other members.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Transactions effected by exchange members through other members. 240.11a2-2(T) Section 240.11a2-2(T) Commodity and Securities... Regulation (rule 11a-1) § 240.11a2-2(T) Transactions effected by exchange members through other members. (a...

17 CFR 240.11a2-2(T) - Transactions effected by exchange members through other members.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 17 Commodity and Securities Exchanges 3 2011-04-01 2011-04-01 false Transactions effected by exchange members through other members. 240.11a2-2(T) Section 240.11a2-2(T) Commodity and Securities... Regulation (rule 11a-1) § 240.11a2-2(T) Transactions effected by exchange members through other members. (a...

Evaluation of seed coating formulations of Trichoderma harzianum on cucumber seeds against pre- and post-emergence damping-off caused by Pythium ultimum

USDA-ARS?s Scientific Manuscript database

Seed coating formulations of Trichoderma harzianum were evaluated on cucumber seeds to control pre- and post-emergence damping-off caused by Pythium ultimum in greenhouse studies. Results showed that coating formulation H reduced the disease incidence significantly, and had the potential for commer...

Control of epithelial cell function by interleukin-22-producing RORγt+ innate lymphoid cells

PubMed Central

Sanos, Stephanie L; Vonarbourg, Cedric; Mortha, Arthur; Diefenbach, Andreas

2011-01-01

It is rapidly emerging that the defence system of innate lymphocytes is more diverse than previously recognized. In addition to natural killer (NK) cells, lymphoid tissue inducer (LTi) cells, and natural helper cells have now been identified. LTi cells are developmentally dependent on the orphan transcription factor RORγt and instruct lymph node development during embryogenesis. More recently, it has become evident, that in addition to their role for lymph organ development, LTi cells are also potent producers of cytokines such as interleukin-22 (IL-22) and IL-17 in adult mice. In addition to LTi cells, another RORγt-dependent innate lymphocyte subset co-expressing RORγt and NK cell receptors (NKRs) has been identified. These NKR+ RORγt+ cells are also potent producers of IL-22 but it is unclear whether they are part of the NK cell or LTi cell lineage. This review will highlight recent progress in understanding development and function of innate IL-22-producing lymphocyte subsets. PMID:21391996

Correlation of structure, function and protein dynamics in GH7 cellobiohydrolases from Trichoderma atroviride, T. reesei and T. harzianum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Borisova, Anna S.; Eneyskaya, Elena V.; Jana, Suvamay

The ascomycete fungus Trichoderma reesei is the predominant source of enzymes for industrial conversion of lignocellulose. Its glycoside hydrolase family 7 cellobiohydrolase (GH7 CBH) TreCel7A constitutes nearly half of the enzyme cocktail by weight and is the major workhorse in the cellulose hydrolysis process. The orthologs from Trichoderma atroviride (TatCel7A) and Trichoderma harzianum (ThaCel7A) show high sequence identity with TreCel7A, ~ 80%, and represent naturally evolved combinations of cellulose-binding tunnel-enclosing loop motifs, which have been suggested to influence intrinsic cellobiohydrolase properties, such as endo-initiation, processivity, and off-rate. The TatCel7A, ThaCel7A, and TreCel7A enzymes were characterized for comparison of function. Themore » catalytic domain of TatCel7A was crystallized, and two structures were determined: without ligand and with thio-cellotriose in the active site. Initial hydrolysis of bacterial cellulose was faster with TatCel7A than either ThaCel7A or TreCel7A. In synergistic saccharification of pretreated corn stover, both TatCel7A and ThaCel7A were more efficient than TreCel7A, although TatCel7A was more sensitive to thermal inactivation. Structural analyses and molecular dynamics (MD) simulations were performed to elucidate important structure/function correlations. Moreover, reverse conservation analysis (RCA) of sequence diversity revealed divergent regions of interest located outside the cellulose-binding tunnel of Trichoderma spp. GH7 CBHs. We hypothesize that the combination of loop motifs is the main determinant for the observed differences in Cel7A activity on cellulosic substrates. Fine-tuning of the loop flexibility appears to be an important evolutionary target in Trichoderma spp., a conclusion supported by the RCA data. Our results indicate that, for industrial use, it would be beneficial to combine loop motifs from TatCel7A with the thermostability features of TreCel7A. Furthermore, one region

Correlation of structure, function and protein dynamics in GH7 cellobiohydrolases from Trichoderma atroviride, T. reesei and T. harzianum

DOE PAGES

Borisova, Anna S.; Eneyskaya, Elena V.; Jana, Suvamay; ...

2018-01-13

The ascomycete fungus Trichoderma reesei is the predominant source of enzymes for industrial conversion of lignocellulose. Its glycoside hydrolase family 7 cellobiohydrolase (GH7 CBH) TreCel7A constitutes nearly half of the enzyme cocktail by weight and is the major workhorse in the cellulose hydrolysis process. The orthologs from Trichoderma atroviride (TatCel7A) and Trichoderma harzianum (ThaCel7A) show high sequence identity with TreCel7A, ~ 80%, and represent naturally evolved combinations of cellulose-binding tunnel-enclosing loop motifs, which have been suggested to influence intrinsic cellobiohydrolase properties, such as endo-initiation, processivity, and off-rate. The TatCel7A, ThaCel7A, and TreCel7A enzymes were characterized for comparison of function. Themore » catalytic domain of TatCel7A was crystallized, and two structures were determined: without ligand and with thio-cellotriose in the active site. Initial hydrolysis of bacterial cellulose was faster with TatCel7A than either ThaCel7A or TreCel7A. In synergistic saccharification of pretreated corn stover, both TatCel7A and ThaCel7A were more efficient than TreCel7A, although TatCel7A was more sensitive to thermal inactivation. Structural analyses and molecular dynamics (MD) simulations were performed to elucidate important structure/function correlations. Moreover, reverse conservation analysis (RCA) of sequence diversity revealed divergent regions of interest located outside the cellulose-binding tunnel of Trichoderma spp. GH7 CBHs. We hypothesize that the combination of loop motifs is the main determinant for the observed differences in Cel7A activity on cellulosic substrates. Fine-tuning of the loop flexibility appears to be an important evolutionary target in Trichoderma spp., a conclusion supported by the RCA data. Our results indicate that, for industrial use, it would be beneficial to combine loop motifs from TatCel7A with the thermostability features of TreCel7A. Furthermore, one region

Integrated options for the management of black root rot of strawberry caused by Rhizoctonia solani Kuhn.

PubMed

Asad-Uz-Zaman, Md; Bhuiyan, Mohammad Rejwan; Khan, Mohammad Ashik Iqbal; Alam Bhuiyan, Md Khurshed; Latif, Mohammad Abdul

2015-02-01

An investigation was made to manage strawberry black root rot caused by Rhizoctonia solani (R. solani) through the integration of Trichoderma harzianum (T. harzianum) isolate STA7, mustard oil cake and Provax 200. A series of preliminary experiments were conducted to select a virulent isolate of R. solani, an effective isolate of T. harzianum, a suitable organic amendment, and a suitable fungicide before setting the experiment for integration. The pathogenicity of the selected four isolates of R. solani was evaluated against strawberry and isolate SR1 was selected as the test pathogen due to its highest virulent (95.47% mortality) characteristics. Among the 20 isolates of T. harzianum, isolate STA7 showed maximum inhibition (71.97%) against the test pathogen (R. solani). Among the fungicides, Provax-200 was found to be more effective at lowest concentration (100 ppm) and highly compatible with Trichoderma isolates STA7. In the case of organic amendments, maximum inhibition (59.66%) of R. solani was obtained through mustard oil cake at the highest concentration (3%), which was significantly superior to other amendments. Minimum percentages of diseased roots were obtained with pathogen (R. solani)+Trichoderma+mustard oil cake+Provax-200 treatment, while the highest was observed with healthy seedlings with a pathogen-inoculated soil. In the case of leaf and fruit rot diseases, significantly lowest infected leaves as well as fruit rot were observed with a pathogen+Trichoderma+mustard oil cake+Provax-200 treatment in comparison with the control. A similar trend of high effectiveness was observed by the integration of Trichoderma, fungicide and organic amendments in controlling root rot and fruit diseases of strawberry. Single application of Trichoderma isolate STA7, Provax 200 or mustard oil cake did not show satisfactory performance in terms of disease-free plants, but when they were applied in combination, the number of healthy plants increased significantly. The

Acquisition of Xyllela fastidiosa causes changes to the inoculation behavior (EPG X wave) of an efficient sharpshooter vector

USDA-ARS?s Scientific Manuscript database

Xylella fastidiosa (Xf) is a foregut-borne bacterium that is inoculated into xylem cells of a healthy plant during feeding by sharpshooter vectors. Inoculation occurs during salivation and egestion behaviors that are likely represented by the sharpshooter X wave. The objective of this study was to t...

Ectopic expression of a novel CD22 splice-variant regulates survival and proliferation in malignant T cells from cutaneous T cell lymphoma (CTCL) patients

PubMed Central

Bagdonaite, Ieva; Wandall, Hans H.; Litvinov, Ivan V.; Nastasi, Claudia; Becker, Jürgen C.; Dabelsteen, Sally; Geisler, Carsten; Bonefeld, Charlotte M.; Zhang, Qian; Wasik, Mariusz A.; Zhou, Youwen; Sasseville, Denis; Ødum, Niels; Woetmann, Anders

2015-01-01

CD22 is a member of the Sialic acid-binding Ig-like lectin (Siglec) family of lectins described to be exclusively present in B lymphocytes and B cell-derived neoplasms. Here, we describe a novel splice form of CD22 (designated CD22ΔN), which lacks the N-terminal domain as demonstrated by exon-specific RT-PCR and differential recognition by anti-CD22 antibodies. Importantly, CD22ΔN mRNA is expressed in skin lesions from 39 out of 60 patients with cutaneous T cell lymphoma (CTCL), whereas few patients (6 out of 60) expresses full-length, wild type CD22 (CD22wt). In addition, IHC staining of tumor biopsies confirmed the expression of CD22 in CD4+ T cells. Moreover, four out of four malignant T cell lines express CD22: Two cell lines express CD22ΔN (MyLa2059 and PB2B) and two express CD22wt (MAC-1 and MAC-2A). siRNA-mediated silencing of CD22 impairs proliferation and survival of malignant T cells, demonstrating a functional role for both CD22ΔN and CD22wt in these cells. In conclusion, we provide the first evidence for an ectopic expression of CD22 and a novel splice variant regulating malignant proliferation and survival in CTCL. Analysis of expression and function of CD22 in cutaneous lymphomas may form the basis for development of novel targeted therapies for our patients. PMID:25957418

Ectopic expression of a novel CD22 splice-variant regulates survival and proliferation in malignant T cells from cutaneous T cell lymphoma (CTCL) patients.

PubMed

Bagdonaite, Ieva; Wandall, Hans H; Litvinov, Ivan V; Nastasi, Claudia; Becker, Jürgen C; Dabelsteen, Sally; Geisler, Carsten; Bonefeld, Charlotte M; Zhang, Qian; Wasik, Mariusz A; Zhou, Youwen; Sasseville, Denis; Ødum, Niels; Woetmann, Anders

2015-06-10

CD22 is a member of the Sialic acid-binding Ig-like lectin (Siglec) family of lectins described to be exclusively present in B lymphocytes and B cell-derived neoplasms. Here, we describe a novel splice form of CD22 (designated CD22∆N), which lacks the N-terminal domain as demonstrated by exon-specific RT-PCR and differential recognition by anti-CD22 antibodies. Importantly, CD22∆N mRNA is expressed in skin lesions from 39 out of 60 patients with cutaneous T cell lymphoma (CTCL), whereas few patients (6 out of 60) expresses full-length, wild type CD22 (CD22wt). In addition, IHC staining of tumor biopsies confirmed the expression of CD22 in CD4+ T cells. Moreover, four out of four malignant T cell lines express CD22: Two cell lines express CD22∆N (MyLa2059 and PB2B) and two express CD22wt (MAC-1 and MAC-2A). siRNA-mediated silencing of CD22 impairs proliferation and survival of malignant T cells, demonstrating a functional role for both CD22∆N and CD22wt in these cells.In conclusion, we provide the first evidence for an ectopic expression of CD22 and a novel splice variant regulating malignant proliferation and survival in CTCL. Analysis of expression and function of CD22 in cutaneous lymphomas may form the basis for development of novel targeted therapies for our patients.

Lead-resistant strain KQBT-3 inoculants of Tricholoma lobayensis Heim that enhance remediation of lead-contaminated soil.

PubMed

Li, Ying; Qin, Chui-Xin; Gao, Biyu; Hu, Yuanjia; Xu, Heng

2015-01-01

To enhance lead-detoxifying efficiency of Tricholoma lobayensis Heim, one lead-resistant strain KQBT-3 (Bacillus thuringiensis) was applied owing to its excellent ability to tolerate Pb. KQBT-3 domesticated in liquid medium with increasing lead concentrations could tolerate Pb(NO3)2 up to a concentration of 800 mg L(-1). Pot experiments showed that the KQBT-3 not only could promote the growth of T. lobayensis, but also could enhance its Pb accumulation ability under heavy metal stress. Biomass and accumulation of Pb increased 47.3% and 33.2%, respectively. In addition, after inoculation of KQBT-3, the significant decrease of malondialdehyde indicated KQBT-3 could alleviate lipid peroxidation in T. lobayensis. What is interesting is that superoxide dismutase and peroxidase activities in T. lobayensis inoculated with KQBT-3 were increased, and the maximum increasing rate was 121.71% and 117.29%, respectively. However, the catalase activity increased slightly. This revealed that inoculating KQBT-3 further induced oxidative response in T. lobayensis due to Pb accumulation. Therefore, the present work showed that KQBT-3 made a major contribution to promote growth and lead uptake of T. lobayensis and alleviate the oxidative stress. This kind of auxiliary effect on macrofungi can be developed into a novel bioremediation strategy.

EOMES-positive CD4+ T cells are increased in PTPN22 (1858T) risk allele carriers.

PubMed

Chemin, Karine; Ramsköld, Daniel; Diaz-Gallo, Lina-Marcela; Herrath, Jessica; Houtman, Miranda; Tandre, Karolina; Rönnblom, Lars; Catrina, Anca; Malmström, Vivianne

2018-04-01

The presence of the PTPN22 risk allele (1858T) is associated with several autoimmune diseases including rheumatoid arthritis (RA). Despite a number of studies exploring the function of PTPN22 in T cells, the exact impact of the PTPN22 risk allele on T-cell function in humans is still unclear. In this study, using RNA sequencing, we show that, upon TCR-activation, naïve human CD4 + T cells homozygous for the PTPN22 risk allele overexpress a set of genes including CFLAR and 4-1BB, which are important for cytotoxic T-cell differentiation. Moreover, the protein expression of the T-box transcription factor Eomesodermin (EOMES) was increased in T cells from healthy donors homozygous for the PTPN22 risk allele and correlated with a decreased number of naïve CD4 + T cells. There was no difference in the frequency of other CD4 + T-cell subsets (Th1, Th17, Tfh, Treg). Finally, an accumulation of EOMES + CD4 + T cells was observed in synovial fluid of RA patients with a more pronounced production of Perforin-1 in PTPN22 risk allele carriers. Altogether, we propose a novel mechanism of action of PTPN22 risk allele through the generation of cytotoxic CD4 + T cells and identify EOMES + CD4 + T cells as a relevant T-cell subset in RA pathogenesis. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Inoculating against reactance to persuasive health messages.

PubMed

Richards, Adam S; Banas, John A

2015-01-01

This investigation examined the possibility of decreasing psychological reactance to health campaigns through the use of inoculation messages. It was hypothesized that an inoculation message, which forewarned of the potential of subsequent reactance, would decrease participants' likelihood of reacting negatively to a freedom-threatening message aimed to reduce excessive alcohol consumption. Participants (N = 275) who were inoculated against potential reactance felt less threatened and experienced less reactance compared to those who did not read an inoculation message. Structural equation modeling showed that inoculation indirectly predicted lower intention to drink alcohol via the theorized mediated reactance process. This research suggests that it is possible to inoculate against self-generated cognitions that might otherwise lead toward negative health behaviors.

Parallel formation and synergism of hydrolytic enzymes and peptaibol antibiotics, molecular mechanisms involved in the antagonistic action of Trichoderma harzianum against phytopathogenic fungi.

PubMed Central

Schirmböck, M; Lorito, M; Wang, Y L; Hayes, C K; Arisan-Atac, I; Scala, F; Harman, G E; Kubicek, C P

1994-01-01

Chitinase, beta-1,3-glucanase, and protease activities were formed when Trichoderma harzianum mycelia, grown on glucose as the sole carbon source, were transferred to fresh medium containing cell walls of Botrytis cinerea. Chitobiohydrolase, endochitinase, and beta-1,3-glucanase activities were immunologically detected in culture supernatants by Western blotting (immunoblotting), and the first two were quantified by enzyme-linked immunosorbent assay. Under the same conditions, exogenously added [U-14C]valine was incorporated in acetone-soluble compounds with an apparent M(r) of < 2,000. These compounds comigrated with the peptaibols trichorzianines A1 and B1 in thin-layer chromatography and released [U-14C]valine after incubation in 6N HCl. Incorporation of radioactive valine into this material was stimulated by the exogenous supply of alpha-aminoisobutyric acid, a rare amino acid which is a major constituent of peptaibols. The obtained culture supernatants inhibited spore germination as well as hyphal elongation of B. cinerea. Culture supernatants from mycelia placed in fresh medium without cell walls of B. cinerea did not show hydrolase activities, incorporation of [U-14C]valine into peptaibol-like compounds, and inhibition of fungal growth. Purified trichorzianines A1 and B1 as well as purified chitobiohydrolase, endochitinase, or beta-1,3-glucanase inhibited spore germination and hyphal elongation, but at concentrations higher than those observed in the culture supernatants. However, when the enzymes and the peptaibols were tested together, an antifungal synergistic interaction was observed and the 50% effective dose values obtained were in the range of those determined in the culture supernatants. Therefore, the parallel formation and synergism of hydrolytic enzymes and antibiotics may have an important role in the antagonistic action of T. harzianum against fungal phytopathogens. Images PMID:7811076

Phosphate Solubilization Potential and Phosphatase Activity Of Rhizospheric Trichoderma Spp.

PubMed Central

Anil, Kapri; Lakshmi, Tewari

2010-01-01

Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P) solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP) broth containing tricalcium phosphate (TCP) as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 εg.ml-1), followed by the standard culture of T. harzianum (386.42 εg.ml-1) after 96 h of incubation at 30±10C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP). High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml-1) followed by the standard culture (13.41 U.ml-1) at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum) growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP). Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg-1 soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth. PMID:24031556

Effect of plant growth-promoting rhizobacteria inoculation on cadmium (Cd) uptake by Eruca sativa.

PubMed

Kamran, Muhammad Aqeel; Syed, Jabir Hussain; Eqani, Syed Ali Musstjab Akber Shah; Munis, Muhammad Farooq Hussain; Chaudhary, Hassan Javed

2015-06-01

Microbe-assisted phyto-remediation approach is widely applied and appropriate choice to reduce the environmental risk of heavy metals originated from contaminated soils. The present study was designed to screen out the nested belongings of Eruca sativa plants and Pseudomonas putida (ATCC 39213) at varying cadmium (Cd) levels and their potential to deal with Cd uptake from soils. We carried out pot trial experiment by examining the soil containing E. sativa seedlings either treated with P. putida and/or untreated plants subjected to three different levels (ppm) of Cd (i.e., 150, 250, and 500). In all studied cases, we observed an increase in Cd uptake for E. sativa plants inoculated with P. putida than those of un-inoculated plants. Cd toxicity was assessed by recording different parameters including stunted shoot growth, poor rooting, and Cd residual levels in the plants that were not inoculated with P. putida. Significant difference (p < 0.05) of different growth parameters for inoculated vs non-inoculated plants was observed at all given treatments. However, among the different treatments, E. sativa exhibited increased values for different growth parameters (except proline contents) at lower Cd levels than those of their corresponding higher levels, shoot length (up to 27 %), root length (up to 32 %), whole fresh plant (up to 40 %), dry weight (up to 22 %), and chlorophyll contents (up to 26 %). Despite the hyperaccumulation of Cd in whole plant of E. sativa, P. putida improved the plant growth at varying levels of Cd supply than those of associated non-inoculated plants. Present results indicated that inoculation with P. putida enhanced the Cd uptake potential of E. sativa and favors the healthy growth under Cd stress.

Chemical consequences of three commercial strains of Oenococcus oeni co-inoculated with Torulaspora delbrueckii in durian wine fermentation.

PubMed

Lu, Yuyun; Chua, Jian-Yong; Huang, Dejian; Lee, Pin-Rou; Liu, Shao-Quan

2017-01-15

This work evaluated for the first time the chemical consequences of three commercial strains of Oenococcus oeni co-inoculated with Torulaspora delbrueckii in durian wine fermentation. Compared with the control (yeast only, 5.70% v/v ethanol produced), samples co-inoculated with T. delbrueckii and O. oeni PN4 improved ethanol production (6.06% v/v), which was significantly higher than samples co-inoculated with Viniflora (4.78% v/v) or Enoferm Beta (5.01% v/v). Wines co-fermented with the respective latter two oenococci contained excessive levels of ethyl acetate (>80mg/L) that were likely to affect negatively wine aroma. In addition, they led to significantly higher acetic and lactic acid production relative to PN4. O. oeni PN4 seemed to be the most suitable strain to co-inoculate with T. delbrueckii for simultaneous alcoholic and malolactic fermentation in durian wine by contributing moderately increased concentrations of higher alcohols, acetate esters and ethyl esters that would have positive sensory impacts. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inhibition of aflatoxin B production of Aspergillus flavus, isolated from soybean seeds by certain natural plant products.

PubMed

Krishnamurthy, Y L; Shashikala, J

2006-11-01

The inhibitory effect of cowdung fumes, Captan, leaf powder of Withania somnifera, Hyptis suaveolens, Eucalyptus citriodora, peel powder of Citrus sinensis, Citrus medica and Punica granatum, neem cake and pongamia cake and spore suspension of Trichoderma harzianum and Aspergillus niger on aflatoxin B(1) production by toxigenic strain of Aspergillus flavus isolated from soybean seeds was investigated. Soybean seed was treated with different natural products and fungicide captan and was inoculated with toxigenic strain of A. flavus and incubated for different periods. The results showed that all the treatments were effective in controlling aflatoxin B(1) production. Captan, neem cake, spore suspension of T. harzianum, A. niger and combination of both reduced the level of aflatoxin B(1) to a great extent. Leaf powder of W. somnifera, H. suaveolens, peel powder of C. sinensis, C. medica and pongamia cake also controlled the aflatoxin B(1) production. All the natural product treatments applied were significantly effective in inhibiting aflatoxin B(1) production on soybean seeds by A. flavus. These natural plant products may successfully replace chemical fungicides and provide an alternative method to protect soybean and other agricultural commodities from aflatoxin B(1) production by A. flavus.

Peptaibol, secondary-metabolite, and hydrophobin pattern of commercial biocontrol agents formulated with species of the Trichoderma harzianum complex.

PubMed

Degenkolb, Thomas; Fog Nielsen, Kristian; Dieckmann, Ralf; Branco-Rocha, Fabiano; Chaverri, Priscila; Samuels, Gary J; Thrane, Ulf; von Döhren, Hans; Vilcinskas, Andreas; Brückner, Hans

2015-04-01

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. Copyright © 2015 Verlag Helvetica Chimica Acta AG, Zürich.

Rhizostabilization of metals in soils using Lupinus luteus inoculated with the metal resistant rhizobacterium Serratia sp. MSMC541.

PubMed

El Aafi, N; Brhada, F; Dary, M; Maltouf, A Filali; Pajuelo, E

2012-03-01

The aim of this work was to test Lupinus luteus plants, inoculated with metal resistant rhizobacteria, in order to phytostabilise metals in contaminated soils. The resistance to heavy metals of strains isolated from nodules of Lupinus plants was evaluated. The strain MSMC541 showed multi-resistance to several metals (up to 13.3 mM As, 2.2 mM Cd, 2.3 mM Cu, 9 mM Pb and 30 mM Zn), and it was selected for further characterization. Furthermore, this strain was able to biosorb great amounts of metals in cell biomass. 16S rDNA sequencing positioned this strain within the genus Serratia. The presence of arsenic resistance genes was confirmed by southern blot and PCR amplification. A rhizoremediation pot experiment was conducted using Lupinus luteus grown on sand supplemented with heavy metals and inoculated with MSMC541. Plant growth parameters and metal accumulation were determined in inoculated vs. non-inoculated Lupinus luteus plants. The results showed that inoculation with MSMC541 improved the plant tolerance to metals. At the same time, metal translocation to the shoot was significantly reduced upon inoculation. These results suggest that Lupinus luteus plants, inoculated with the metal resistant strain Serratia sp. MSMC541, have a great potential for phytostabilization of metal contaminated soils.

Mineral Soils as Carriers for Rhizobium Inoculants

PubMed Central

Chao, W.-L.; Alexander, Martin

1984-01-01

Mineral soil-based inoculants of Rhizobium meliloti and Rhizobium phaseoli survived better at 4°C than at higher temperatures, but ca. 15% of the cells were viable at 37°C after 27 days. Soil-based inoculants of R. meliloti, R. phaseoli, Rhizobium japonicum, and a cowpea Rhizobium sp. applied to seeds of their host legumes also survived better at low temperatures, but the percent survival of such inoculants was higher than peat-based inoculants at 35°C. Survival of R. phaseoli, R. japonicum, and cowpea rhizobia was not markedly improved when the cells were suspended in sugar solutions before drying them in soil. Nodulation was abundant on Phaseolus vulgaris derived from seeds that had been coated with a soil-based inoculant and stored for 165 days at 25°C. The increase in yield and nitrogen content of Phaseolus angularis grown in the greenhouse was the same with soil-and peat-based inoculants. We suggest that certain mineral soils can be useful and readily available carriers for legume inoculants containing desiccation-resistant Rhizobium strains. PMID:16346460

Soil inoculation with symbiotic microorganisms promotes plant growth and nutrient transporter genes expression in durum wheat

PubMed Central

Saia, Sergio; Rappa, Vito; Ruisi, Paolo; Abenavoli, Maria Rosa; Sunseri, Francesco; Giambalvo, Dario; Frenda, Alfonso S.; Martinelli, Federico

2015-01-01

In a field experiment conducted in a Mediterranean area of inner Sicily, durum wheat was inoculated with plant growth-promoting rhizobacteria (PGPR), with arbuscular mycorrhizal fungi (AMF), or with both to evaluate their effects on nutrient uptake, plant growth, and the expression of key transporter genes involved in nitrogen (N) and phosphorus (P) uptake. These biotic associations were studied under either low N availability (unfertilized plots) and supplying the soil with an easily mineralizable organic fertilizer. Regardless of N fertilization, at the tillering stage, inoculation with AMF alone or in combination with PGPR increased the aboveground biomass yield compared to the uninoculated control. Inoculation with PGPR enhanced the aboveground biomass yield compared to the control, but only when N fertilizer was added. At the heading stage, inoculation with all microorganisms increased the aboveground biomass and N. Inoculation with PGPR and AMF+PGPR resulted in significantly higher aboveground P compared to the control and inoculation with AMF only when organic N was applied. The role of microbe inoculation in N uptake was elucidated by the expression of nitrate transporter genes. NRT1.1, NRT2, and NAR2.2 were significantly upregulated by inoculation with AMF and AMF+PGPR in the absence of organic N. A significant down-regulation of the same genes was observed when organic N was added. The ammonium (NH4+) transporter genes AMT1.2 showed an expression pattern similar to that of the NO3- transporters. Finally, in the absence of organic N, the transcript abundance of P transporters Pht1 and PT2-1 was increased by inoculation with AMF+PGPR, and inoculation with AMF upregulated Pht2 compared to the uninoculated control. These results indicate the soil inoculation with AMF and PGPR (alone or in combination) as a valuable option for farmers to improve yield, nutrient uptake, and the sustainability of the agro-ecosystem. PMID:26483827

Overexpression of erg1 gene in Trichoderma harzianum CECT 2413: effect on the induction of tomato defence-related genes.

PubMed

Cardoza, R E; Malmierca, M G; Gutiérrez, S

2014-09-01

To investigate the effect of the overexpression of erg1 gene of Trichoderma harzianum CECT 2413 (T34) on the Trichoderma-plant interactions and in the biocontrol ability of this fungus. Transformants of T34 strain overexpressing erg1 gene did not show effect on the ergosterol level, although a drastic decrease in the squalene level was observed in the transformants at 96 h of growth. During interaction with plants, the erg1 overexpression resulted in a reduction of the priming ability of several tomato defence-related genes belonging to the salicylate pathway, and also of the TomLoxA gene, which is related to the jasmonate pathway. Interestingly, other jasmonate-related genes, such as PINI and PINII, were slightly induced. The erg1 overexpressed transformants also showed a reduced ability to colonize tomato roots. The ergosterol biosynthetic pathway might play an important role in regulating Trichoderma-plant interactions, although this role does not seem to be restricted to the final product; instead, other intermediates such as squalene, whose role in the Trichoderma-plant interaction has not been characterized, would also play an important role. The functional analysis of genes involved in the synthesis of ergosterol could provide additional strategies to improve the ability of biocontrol of the Trichoderma strains and their interaction with plants. © 2014 The Society for Applied Microbiology.

Simultaneous occurrence of t(9;22)(q34;q11.2) and t(16;16)(p13;q22) in a patient with chronic myeloid leukemia in blastic phase.

PubMed

Zámecníkova, Adriana; Al Bahar, Soad; Ramesh, Pandita

2008-06-01

Coexistence of two specific chromosomal translocations in the same clone is an infrequent phenomenon and has only rarely been reported in hematological malignancies. We report a combination of t(16;16)(p13;q22), the Philadelphia translocation t(9;22)(q34;q11.2), and deletion of the long arm of chromosome 7 in a patient with chronic myeloid leukemia in blast phase. Monotherapy treatment with imatinib mesylate resulted in the disappearance of the Ph-positive clone, but with persistence of t(16;16) and del(7) in all of the metaphases examined. The case illustrates that, although imatinib mesylate can be an effective treatment in eradication of the BCR-ABL fusion gene cells, the occurrence of additional specific abnormalities in Philadelphia-positive leukemias may pose a significant therapeutic challenge. (c) 2008 Elsevier Inc.

Intrauterine inoculation of minipigs with Chlamydia trachomatis during diestrus establishes a longer lasting infection compared to vaginal inoculation during estrus.

PubMed

Lorenzen, Emma; Follmann, Frank; Secher, Jan O; Goericke-Pesch, Sandra; Hansen, Mette S; Zakariassen, Hannah; Olsen, Anja W; Andersen, Peter; Jungersen, Gregers; Agerholm, Jørgen S

2017-06-01

Advanced animal models, such as minipigs, are needed for the development of a globally requested human Chlamydia vaccine. Previous studies have shown that vaginal inoculation of sexually mature Göttingen minipigs with Chlamydia trachomatis resulted in an infection lasting only 3-5 days. The aim of this study was to evaluate the effect of targeting the upper porcine genital tract by transcervical and transabdominal intrauterine inoculation, compared to previously performed vaginal inoculation. Furthermore, we investigated the effect of the hormonal cycle, estrus vs. diestrus, on the establishment of a C. trachomatis infection in the minipig. Targeting the upper genital tract (transcervical inoculation) resulted in a longer lasting infection (at least 7 days) compared to vaginal inoculation (3-5 days). When comparing intrauterine inoculation during estrus and diestrus, inoculation during diestrus resulted in a longer lasting infection (at least 10 days) compared to estrus (3-5 days). Furthermore, we found a significant C. trachomatis specific IFN-γ response in pigs inoculated during estrus correlating with the accelerated clearance of infection in these pigs. These findings suggest that for implementation of an optimal model of C. trachomatis in minipigs, inoculation should bypass the cervix and preferable be performed during diestrus. Copyright © 2017 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.

A dry-inoculation method for nut kernels.

PubMed

Blessington, Tyann; Theofel, Christopher G; Harris, Linda J

2013-04-01

A dry-inoculation method for almonds and walnuts was developed to eliminate the need for the postinoculation drying required for wet-inoculation methods. The survival of Salmonella enterica Enteritidis PT 30 on wet- and dry-inoculated almond and walnut kernels stored under ambient conditions (average: 23 °C; 41 or 47% RH) was then compared over 14 weeks. For wet inoculation, an aqueous Salmonella preparation was added directly to almond or walnut kernels, which were then dried under ambient conditions (3 or 7 days, respectively) to initial nut moisture levels. For the dry inoculation, liquid inoculum was mixed with sterilized sand and dried for 24 h at 40 °C. The dried inoculated sand was mixed with kernels, and the sand was removed by shaking the mixture in a sterile sieve. Mixing procedures to optimize the bacterial transfer from sand to kernel were evaluated; in general, similar levels were achieved on walnuts (4.8-5.2 log CFU/g) and almonds (4.2-5.1 log CFU/g). The decline of Salmonella Enteritidis populations was similar during ambient storage (98 days) for both wet-and dry-inoculation methods for both almonds and walnuts. The dry-inoculation method mimics some of the suspected routes of contamination for tree nuts and may be appropriate for some postharvest challenge studies. Copyright © 2012 Elsevier Ltd. All rights reserved.

Screening of bioagents against root rot of mung bean caused by Rhizoctonia solani.

PubMed

Singh, Surender; Chand, Hari

2006-01-01

A laboratory and green house experiment was carried out on the comparative antagonistic performance of four different bioagents (Aspergillus sp. Gliocladium virens, Trichoderma harzianum and T. viride) isolated from soil against Rhizoctonia solani. Under laboratory conditions, T. harzianum exhibited maximum (75.55%) mycelial growth inhibition of R. solani. This was followed by T. viride, which showed 65.93% mycelial growth inhibition of the pathogen. Gliocladium virens was also found to be effective antagonists, which exhibited 57.77% mycelial growth inhibition. While Aspergillus sp exhibited minimum growth inhibition (45.74%) in comparison to other bioagents. Under green house conditions, T. harzianum gave maximum protection of the disease (72.72%) followed by T. viride, which exhibited 54.54% disease control. However, G. virens and Aspergillus sp were found least effective in controlling root rot of mungbean.

PTPN22 1858C > T polymorphism and susceptibility to systemic lupus erythematosus: a meta-analysis update.

PubMed

de Lima, Suelen Cristina; Adelino, José Eduardo; Crovella, Sergio; de Azevedo Silva, Jaqueline; Sandrin-Garcia, Paula

2017-11-01

Studies performed in the past years showed PTNP22 1858 C > T (rs2476601) polymorphism as associated with systemic lupus erythematosus susceptibility, although conflicting findings are still found. In this context, a powerful statistical study, such as meta-analysis, is necessary to establish a consensus. The aim of this study was to evaluate association studies between the PTPN22 1858 C > T polymorphism and SLE by a meta-analysis update, including three recently published studies in the last three years. A total of 3868 SLE patients and 7458 healthy individuals were considered herein, enclosing 19 studies from Asian, American, European and Latin ethnic groups. Odds ratio (OR) was performed for allelic, dominant and recessive genetic models. Statistically significant association was found between the PTPN22 1858 C > T polymorphism and susceptibility to SLE in all inheritance models. Allelic genetic model data (OR = 1.54, 95% confidence interval (CI) = 1.38-1.72, p value=.000) shows that T allele confers increased SLE susceptibility. As well as recessive genetic model (OR = 2.04, 95% CI = 1.09-3.82, p value = .030) for T/T genotype. Instead, dominant genetic model shows that C/C genotype confers lower susceptibility for SLE development (OR = 0.62, 95% CI = 0.54-0.72, p value = .000). In addition, we provided an ethnicity-derived meta-analysis. The results showed association in Caucasian (OR = 1.47, p value = .000) and Latin (OR = 2.41, p value = .000) ethnic groups. However, rs2476601 polymorphism is not associated nor in Asian (OR= 1.31; p value = .54) and African (OR = 2.04; p value=.22) populations. In conclusion, present meta-analysis update confirms that T allele and T/T genotype in PTPN22 1858 C > T polymorphism confers SLE susceptibility, particular in Caucasian and Latin groups, suggesting PTPN22 1858 C > T as a potential genetic marker in SLE susceptibility.

Cotransformation of Trichoderma harzianum with β-Glucuronidase and Green Fluorescent Protein Genes Provides a Useful Tool for Monitoring Fungal Growth and Activity in Natural Soils†

PubMed Central

Bae, Yeoung-Seuk; Knudsen, Guy R.

2000-01-01

Trichoderma harzianum was cotransformed with genes encoding green fluorescent protein (GFP), β-glucuronidase (GUS), and hygromycin B (hygB) resistance, using polyethylene glycol-mediated transformation. One cotransformant (ThzID1-M3) was mitotically stable for 6 months despite successive subculturing without selection pressure. ThzID1-M3 morphology was similar to that of the wild type; however, the mycelial growth rate on agar was reduced. ThzID1-M3 was formed into calcium alginate pellets and placed onto buried glass slides in a nonsterile soil, and its ability to grow, sporulate, and colonize sclerotia of Sclerotinia sclerotiorum was compared with that of the wild-type strain. Wild-type and transformant strains both colonized sclerotia at levels above those of indigenous Trichoderma spp. in untreated controls. There were no significant differences in colonization levels between wild-type and cotransformant strains; however, the presence of the GFP and GUS marker genes permitted differentiation of introduced Trichoderma from indigenous strains. GFP activity was a useful tool for nondestructive monitoring of the hyphal growth of the transformant in a natural soil. The green color of cotransformant hyphae was clearly visible with a UV epifluorescence microscope, while indigenous fungi in the same samples were barely visible. Green-fluorescing conidiophores and conidia were observed within the first 3 days of incubation in soil, and this was followed by the formation of terminal and intercalary chlamydospores and subsequent disintegration of older hyphal segments. Addition of 5-bromo-4-chloro-3-indolyl-β-d-glucuronic acid (X-Gluc) substrate to recovered glass slides confirmed the activity of GUS as well as GFP in soil. Our results suggest that cotransformation with GFP and GUS can provide a valuable tool for the detection and monitoring of specific strains of T. harzianum released into the soil. PMID:10653755

Effect of Inoculant Alloy Selection and Particle Size on Efficiency of Isomorphic Inoculation of Ti-Al

PubMed Central

Rouat, Bernard; Daloz, Dominique; Bouzy, Emmanuel

2018-01-01

The process of isomorphic inoculation relies on precise selection of inoculant alloys for a given system. Three alloys, Ti-10Al-25Nb, Ti-25Al-10Ta, and Ti-47Ta (at %) were selected as potential isomorphic inoculants for a Ti-46Al alloy. The binary Ti-Ta alloy selected was found to be ineffective as an inoculant due to its large density difference with the melt, causing the particles to settle. Both ternary alloys were successfully implemented as isomorphic inoculants that decreased the equiaxed grain size and increased the equiaxed fraction in their ingots. The degree of grain refinement obtained was found to be dependent on the number of particles introduced to the melt. Also, more new grains were formed than particles added to the melt. The grains/particle efficiency varied from greater than one to nearly twenty as the size of the particle increased. This is attributed to the breaking up of particles into smaller particles by dissolution in the melt. For a given particle size, Ti-Al-Ta and Ti-Al-Nb particles were found to have a roughly similar grain/particle efficiency. PMID:29693591

Detecting the Hormonal Pathways in Oilseed Rape behind Induced Systemic Resistance by Trichoderma harzianum TH12 to Sclerotinia sclerotiorum.

PubMed

Alkooranee, Jawadayn Talib; Aledan, Tamarah Raad; Ali, Ali Kadhim; Lu, Guangyuan; Zhang, Xuekun; Wu, Jiangsheng; Fu, Chunhua; Li, Maoteng

2017-01-01

Plants have the ability to resist pathogen attack after infection or treatment with biotic and abiotic elicitors. In oilseed rape plant Brassica napus AACC and in the artificially synthesized Raphanus alboglabra RRCC, the root-colonizing Trichoderma harzianum TH12 fungus triggers induced systemic resistance (ISR), and its culture filtrate (CF) triggers a systemic acquired resistance (SAR) response against infection by the Sclerotinia sclerotiorum. Salicylic acid (SA) and jasmonic acid/ethylene (JA/ET) are plant hormone signals that play important roles in the regulation of ISR and SAR. In this study, at six different time points (1, 2, 4, 6, 8 and 10 days post-infection [dpi]), six resistance genes were used as markers of signaling pathways: JA/ET signaling used AOC3, PDF1.2 and ERF2 genes, while PR-1, TGA5 and TGA6 genes were used as markers of SA signaling. The results of quantitative real-time polymerase chain reaction (qRT-PCR) showed that AOC3, PDF1.2 and ERF2 expression levels in infected leaves of AACC and RRCC increase at 1 and 2 dpi with S. sclerotiorum or inoculation with TH12. PR-1, TGA5 and TGA6 expression levels increased at 8 and 10 dpi in infected leaves. PR-1, TGA5 and TGA6 expression levels increased early in plants treated with CF in both of the healthy genotypes. Furthermore, induction of SA- and JA/ET-dependent defense decreased disease symptoms in infected leaves at different times. The results suggest that the RRCC genotype exhibits resistance to disease and that the ability of TH12 and its CF to induce systemic resistance in susceptible and resistant oilseed rape genotypes exists. In addition, the results indicate for the first time that in RRCC the SA signaling pathway is involved in resistance to necrotrophic pathogens.

Changes in rumen bacterial community composition following feeding of silage inoculated with a commercial silage inoculant

USDA-ARS?s Scientific Manuscript database

Some silage inoculants yield an increase in milk production without increasing fiber digestibility, possibly through altering the rumen microflora. We hypothesized that silage treated with a commercial inoculant (Lactobacillus plantarum, LP) would improve milk production and would alter rumen bacter...

Meiotic Recombination and Spatial Proximity in the Etiology of the Recurrent t(11;22)

PubMed Central

Ashley, Terry; Gaeth, Ann P.; Inagaki, Hidehito; Seftel, Allen; Cohen, Maimon M.; Anderson, Lorinda K.; Kurahashi, Hiroki; Emanuel, Beverly S.

2006-01-01

Although balanced translocations are among the most common human chromosomal aberrations, the constitutional t(11;22)(q23;q11) is the only known recurrent non-Robertsonian translocation. Evidence indicates that de novo formation of the t(11;22) occurs during meiosis. To test the hypothesis that spatial proximity of chromosomes 11 and 22 in meiotic prophase oocytes and spermatocytes plays a role in the rearrangement, the positions of the 11q23 and 22q11 translocation breakpoints were examined. Fluorescence in situ hybridization with use of DNA probes for these sites demonstrates that 11q23 is closer to 22q11 in meiosis than to a control at 6q26. Although chromosome 21p11, another control, often lies as close to 11q23 as does 22q11 during meiosis, chromosome 21 rarely rearranges with 11q23, and the DNA sequence of chromosome 21 appears to be less susceptible than 22q11 to double-strand breaks (DSBs). It has been suggested that the rearrangement recurs as a result of the palindromic AT-rich repeats at both 11q23 and 22q11, which extrude hairpin structures that are susceptible to DSBs. To determine whether the DSBs at these sites coincide with normal hotspots of meiotic recombination, immunocytochemical mapping of MLH1, a protein involved in crossing over, was employed. The results indicate that the translocation breakpoints do not coincide with recombination hotspots and therefore are unlikely to be the result of meiotic programmed DSBs, although MRE11 is likely to be involved. Previous analysis indicated that the DSBs appear to be repaired by a mechanism similar to nonhomologous end joining (NHEJ), although NHEJ is normally suppressed during meiosis. Taken together, these studies support the hypothesis that physical proximity between 11q23 and 22q11—but not typical meiotic recombinational activity in meiotic prophase—plays an important role in the generation of the constitutional t(11;22) rearrangement. PMID:16909390

Assessment of the rhizosphere competency and pentachlorophenol-metabolizing activity of a pesticide-degrading strain of Trichoderma harzianum introduced into the root zone of corn seedlings.

PubMed

Rigot, Jerĵme; Matsumura, Fumio

2002-05-01

To develop a dependable approach to introduce laboratory selected, pesticide-degrading microorganisms into soil environments for the purpose of in situ bioremediation, we tested the possibility of utilizing plant rhizospheres as the vehicle. We first established the rhizosphere competency of a strain of the soil fungus Trichoderma harzianum, a biocontrol species well studied by plant pathologists to colonize plant rhizosphere in many parts of the world. The strain we chose, T.h.2023 is resistant to many fungicides, and it has been shown to metabolize several pesticides. Second, we found that it readily metabolized pentachlorophenol (PCP), which is quickly and stochiometrically converted to pentachloroanisole (PCA) in liquid culture. Taking advantage of this specific feature, we have developed a sensitive metabolic marker approach that allowed us to monitor for the continuous presence and activity of this fungal strain in the corn rhizosphere soil in situ over time.

Chitinase Expression Due to Reduction in Fusaric Acid Level in an Antagonistic Trichoderma harzianum S17TH.

PubMed

Sharma, Vivek; Bhandari, Pamita; Singh, Bikram; Bhatacharya, Amita; Shanmugam, Veerubommu

2013-06-01

To study the effect of reduction in phytotoxin level on fungal chitinases, antagonistic Trichoderma spp. were screened for their ability to reduce the level of fusaric acid (FA), the phytotoxin produced by Fusarium spp. A T. harzianum isolate S17TH was able to tolerate high levels of FA (up to 500 ppm) but was unable to reduce the toxin to a significant level (non-toxic) added to minimal synthetic broth (MSB). However, the isolate was able to reduce 400 ppm FA in the liquid medium after 7 days to a non-toxic level and displayed similar level of antagonism over the control (without FA). In studies of the effect of the reduction in FA (400 ppm) level on chitinase gene expression in PCR assays, nag1 was significantly repressed but ech42 expression was only slightly repressed. Chitinase activity was either reduced or absent in the extracellular proteins of MSB supplemented with 400 ppm FA, which could be attributed to the effect of residual FA or its breakdown products through unknown mechanisms. Selection of S17TH as a toxin insensitive isolate that could commensurate the negative effect on chitinase activity makes it a potential antagonist against Fusarium spp.

Mechanisms of Rapid Induction of Interleukin-22 in Activated T Cells and Its Modulation by Cyclosporin A*

PubMed Central

Rudloff, Ina; Bachmann, Malte; Pfeilschifter, Josef; Mühl, Heiko

2012-01-01

IL-22 is an immunoregulatory cytokine displaying pathological functions in models of autoimmunity like experimental psoriasis. Understanding molecular mechanisms driving IL-22, together with knowledge on the capacity of current immunosuppressive drugs to target this process, may open an avenue to novel therapeutic options. Here, we sought to characterize regulation of human IL22 gene expression with focus on the established model of Jurkat T cells. Moreover, effects of the prototypic immunosuppressant cyclosporin A (CsA) were investigated. We report that IL-22 induction by TPA/A23187 (T/A) or αCD3 is inhibited by CsA or related FK506. Similar data were obtained with peripheral blood mononuclear cells or purified CD3+ T cells. IL22 promoter analysis (−1074 to +156 bp) revealed a role of an NF-AT (−95/−91 nt) and a CREB (−194/−190 nt) binding site for gene induction. Indeed, binding of CREB and NF-ATc2, but not c-Rel, under the influence of T/A to those elements could be proven by ChIP. Because CsA has the capability to impair IκB kinase (IKK) complex activation, the IKKα/β inhibitor IKKVII was evaluated. IKKVII likewise reduced IL-22 induction in Jurkat cells and peripheral blood mononuclear cells. Interestingly, transfection of Jurkat cells with siRNA directed against IKKα impaired IL22 gene expression. Data presented suggest that NF-AT, CREB, and IKKα contribute to rapid IL22 gene induction. In particular the crucial role of NF-AT detected herein may form the basis of direct action of CsA on IL-22 expression by T cells, which may contribute to therapeutic efficacy of the drug in autoimmunity. PMID:22170067

Cast Iron Inoculation Enhanced by Supplementary Oxy-sulfides Forming Elements

NASA Astrophysics Data System (ADS)

Riposan, Iulian; Stan, Stelian; Uta, Valentin; Stefan, Ion

2017-09-01

Inoculation is one of the most important metallurgical treatments applied to the molten cast iron immediately prior to casting, to promote solidification without excessive eutectic undercooling, which favors carbides formation usually with undesirable graphite morphologies. The paper focused on the separate addition of an inoculant enhancer alloy [S, O, oxy-sulfides forming elements] with a conventional Ca-FeSi alloy, in the production of gray and ductile cast irons. Carbides formation tendency decreased with improved graphite characteristics as an effect of the [Ca-FeSi + Enhancer] inoculation combination, when compared to other Ca/Ca, Ba/Ca, RE-FeSi alloy treatments. Adding an inoculant enhancer greatly enhances inoculation, lowers inoculant consumption up to 50% or more and avoids the need to use more costly inoculants, such as a rare earth bearing alloy. The Inoculation Specific Factor [ISF] was developed as a means to more realistically measure inoculant treatment efficiency. It compares the ratio between the improved characteristic level and total inoculant consumption for this effect. Addition of any of the commercial inoculants plus the inoculant enhancer offered outstanding inoculation power [increased ISF] even at higher solidification cooling rates, even though the total enhancer addition was at a small fraction of the amount of commercial inoculant used.

Induction of the immune response suppression in mice inoculated with Candida albicans.

PubMed

Valdez, J C; Mesón, D E; Sirena, A; de Petrino, S F; Eugenia, M; de Jorrat, B B; de Valdex, M G

1986-03-01

There is a controversy in respect to the immunological response (humoral or cellular) concerning the defense against Candida albicans. Candidosis would induce sub-populations of suppressor cells in the host cell-immune response. This report tries to show the effect of different doses of C. albicans (alive or heat-killed) on the expression of cell-mediated and humoral immunity. The effect upon cell immunity was determined by inoculating different lots of singeneic mice, doses of varied concentration of C. albicans and checking for delayed-type hipersensitivity (D.T.H.). D.T.H. was also controlled in syngeneic normal mice which had previously been injected with inoculated mice spleen cells. Humoral immunity was assayed by measuring the induced blastogenesis by Pokeweed Mitogen on spleen mononuclear cells with different doses of C. albicans. Results obtained show that the different doses gave origin to: Suppression of humoral and cell response (10(8) alive); Suppression of only humoral response (10(6) alive); Suppression of cell response and increase of humoral response (10(9) dead); Increase of both responses (10(8) dead).

Genetic diversity of symbiotic Bradyrhizobium elkanii populations recovered from inoculated and non-inoculated Acacia mangium field trials in Brazil.

PubMed

Perrineau, M M; Le Roux, C; de Faria, S M; de Carvalho Balieiro, F; Galiana, A; Prin, Y; Béna, G

2011-07-01

Acacia mangium is a legume tree native to Australasia. Since the eighties, it has been introduced into many tropical countries, especially in a context of industrial plantations. Many field trials have been set up to test the effects of controlled inoculation with selected symbiotic bacteria versus natural colonization with indigenous strains. In the introduction areas, A. mangium trees spontaneously nodulate with local and often ineffective bacteria. When inoculated, the persistence of inoculants and possible genetic recombination with local strains remain to be explored. The aim of this study was to describe the genetic diversity of bacteria spontaneously nodulating A. mangium in Brazil and to evaluate the persistence of selected strains used as inoculants. Three different sites, several hundred kilometers apart, were studied, with inoculated and non-inoculated plots in two of them. Seventy-nine strains were isolated from nodules and sequenced on three housekeeping genes (glnII, dnaK and recA) and one symbiotic gene (nodA). All but one of the strains belonged to the Bradyrhizobium elkanii species. A single case of housekeeping gene transfer was detected among the 79 strains, suggesting an extremely low rate of recombination within B. elkanii, whereas the nodulation gene nodA was found to be frequently transferred. The fate of the inoculant strains varied depending on the site, with a complete disappearance in one case, and persistence in another. We compared our results with the sister species Bradyrhizobium japonicum, both in terms of population genetics and inoculant strain destiny. Copyright © 2011 Elsevier GmbH. All rights reserved.

Accessing inoculation methods of maize and wheat with Azospirillum brasilense.

PubMed

Fukami, Josiane; Nogueira, Marco Antonio; Araujo, Ricardo Silva; Hungria, Mariangela

2016-03-01

The utilization of inoculants containing Azospirillum is becoming more popular due to increasing reports of expressive gains in grain yields. However, incompatibility with pesticides used in seed treatments represents a main limitation for a successful inoculation. Therefore, in this study we searched for alternatives methods for seed inoculation of maize and wheat, aiming to avoid the direct contact of bacteria with pesticides. Different doses of inoculants containing Azospirillum brasilense were employed to perform inoculation in-furrow, via soil spray at sowing and via leaf spray after seedlings had emerged, in comparison to seed inoculation. Experiments were conducted first under greenhouse controlled conditions and then confirmed in the field at different locations in Brazil. In the greenhouse, most parameters measured responded positively to the largest inoculant dose used in foliar sprays, but benefits could also be observed from both in-furrow and soil spray inoculation. However, our results present evidence that field inoculation with plant-growth promoting bacteria must consider inoculant doses, and point to the need of fine adjustments to avoid crossing the threshold of growth stimulation and inhibition. All inoculation techniques increased the abundance of diazotrophic bacteria in plant tissues, and foliar spray improved colonization of leaves, while soil inoculations favored root and rhizosphere colonization. In field experiments, inoculation with A. brasilense allowed for a 25 % reduction in the need for N fertilizers. Our results have identified alternative methods of inoculation that were as effective as the standard seed inoculation that may represent an important strategy to avoid the incompatibility between inoculant bacteria and pesticides employed for seed treatment.

Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches' broom disease

PubMed Central

De Marco, Janice L; Felix, Carlos Roberto

2002-01-01

Background Several Trichoderma strains have been reported to be effective in controlling plant diseases, and the action of fungal hydrolytic enzymes has been considered as the main mechanism involved in the antagonistic process. However, although Trichoderma strains were found to impair development of Crinipellis perniciosa, the causal agent of cocoa plant witches' broom disease, no fungal strain is available for effective control of this disease. We have then undertaken a program of construction of hydrolytic enzyme-overproducing Trichoderma strains aiming improvement of the fungal antagonistic capacity. The protease of an indian Trichoderma isolate showing antagonistic activity against C. perniciosa was purified to homogeneity and characterized for its kinetic properties and action on the phytopathogen cell wall. Results A protease produced by the Trichoderma harzianum isolate 1051 was purified to homogeneity by precipitation with ammonium sulfate followed by hydrophobic chromatography. The molecular mass of this protease as determined by SDS-polyacrylamide gel electrophoresis was about 18.8 kDa. Its N-terminal amino acid sequence shares no homology with any other protease. The purified enzyme substantially affected the cell wall of the phytopathogen C. perniciosa. Western-blotting analysis showed that the enzyme was present in the culture supernatant 24 h after the Trichoderma started to grow in casein-containing liquid medium. Conclusions The capacity of the Trichoderma harzianum protease to hydrolyze the cell wall of C. perniciosa indicates that this enzyme may be actually involved in the antagonistic process between the two fungi. This fact strongly suggest that hydrolytic enzyme over-producing transgenic fungi may show superior biocontrol capacity. PMID:11835696

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation.

PubMed

Li, Ya-Qian; Song, Kai; Li, Ya-Chai; Chen, Jie

2016-08-01

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×10(8) spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×10(8) spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents.

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation*

PubMed Central

Li, Ya-qian; Song, Kai; Li, Ya-chai; Chen, Jie

2016-01-01

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×108 spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×108 spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents. PMID:27487807

Topical Administration Is a Promising Inoculating Route versus Intramuscular Inoculation for the Nanoparticle-Carried DNA Vaccine to Prevent Corneal Infections.

PubMed

Hu, Kai; Malla, Tejsu; Zhai, Yujia; Dong, Lili; Tang, Ru

2015-01-01

To evaluate the comparative effect of topical versus intramuscular administration of nanoparticle-carried DNA vaccine in preventing corneal herpes simplex virus type 1 (HSV-1) infection. Nanoparticle [polyethylenimine (PEI)-Fe3O4]-carried DNA vaccine (PEI-Fe3O4-pRSC-gD-IL-21) or DNA vaccine (pRSC-gD-IL-21) alone were topically versus intramuscularly inoculated into one eye each of mice on days 0, 14 and 28. Three weeks after the final immunization, the specific immune responses and clinical degrees of primary herpes simplex keratitis were evaluated. Topical inoculation of nanoparticle-carried DNA vaccine induced mice to generate similar levels of specific HSV-1-neutralizing antibody, IFN-γ and IL-4 in serum and specific killing (cytotoxicity) and proliferative activities of the splenic lymphocytes, but a significantly higher level of secretory IgA in tears compared to those of intramuscular inoculation. More importantly, the mice inoculated topically showed a significantly decreased herpes simplex keratitis severity than the mice inoculated intramuscularly after HSV-1 challenge on the corneas of the mice. Topical inoculation of nanoparticle-carried DNA vaccine elicits a stronger specific local immune response and more effectively inhibits herpes simplex keratitis as compared to intramuscular inoculation in an HSV-1 ocular challenge mouse model. Thus, topical administration may be a promising inoculating route for the nanoparticle-carried DNA vaccine to prevent corneal infections. © 2015 S. Karger AG, Basel.

Removal of Calcium from Scheelite Leaching Solution by Addition of CaSO4 Inoculating Crystals

NASA Astrophysics Data System (ADS)

Liu, Wenting; Li, Yongli; Zeng, Dewen; Li, Jiangtao; Zhao, Zhongwei

2018-04-01

In this work, the solubility behaviors of gypsum and anhydrite in the H2SO4-H3PO4-H2O system were investigated over the temperature range T = 30-80°C, and the results showed that the solubility of anhydrite was considerably lower than that of gypsum. On the basis of the differential solubilities of gypsum and anhydrite, a method was developed to remove calcium from the scheelite leaching solution by adding anhydrite as an inoculating crystal. The effects of the reaction time, concentration of the CaSO4 inoculating crystals, and temperature were investigated. With an addition of CaSO4 inoculating crystals at a concentration of 60 g/L, the Ca2+ concentration of the scheelite leaching solution decreased to a low level of approximately 0.76 g/L after 10 h at 70°C.

Skin-infiltrating, interleukin-22-producing T cells differentiate pediatric psoriasis from adult psoriasis.

PubMed

Cordoro, Kelly M; Hitraya-Low, Maria; Taravati, Keyon; Sandoval, Priscila Munoz; Kim, Esther; Sugarman, Jeffrey; Pauli, Mariela L; Liao, Wilson; Rosenblum, Michael D

2017-09-01

Evidence from adult psoriasis studies implicates an imbalance between regulatory and effector T cells, particularly T H -17-producing T cells, in the pathogenesis of psoriasis. Little is known about the immunopathology of psoriasis in children. We sought to functionally characterize the inflammatory cell profiles of psoriatic plaques from pediatric patients and compare them with healthy, age-matched controls and adult psoriasis patients. Skin samples from pediatric psoriasis patients and healthy controls were analyzed by multiparameter flow cytometry to determine the dominant immune cell subsets present and cytokines produced. Lesional tissue from pediatric psoriasis patients had significantly increased interleukin (IL) 22 derived from CD4 + and CD8 + cells compared with the tissues from healthy pediatric controls and adult psoriasis patients. Tissue from pediatric psoriasis patients had significantly less elevation of IL-17 derived from CD4 + and CD8 + cells compared with the tissue from adult psoriasis patients. In contrast with the lesions from adult patients, lesional skin in pediatric patients with psoriasis did not have increases in regulatory T cells. This is a pilot study, thus the sample size is small. Significant differences in IL-17 and IL-22 expression were observed in the pediatric psoriasis patients compared with pediatric healthy controls and adult psoriasis patients. IL-22 might be relevant in the pathogenesis of pediatric psoriasis and represents a potential treatment target unique to pediatric psoriasis. Copyright © 2017 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.

Biological control of Botrytis gray mould on tomato cultivated in greenhouse.

PubMed

Fiume, F; Fiume, G

2006-01-01

Research was carried out to evaluate the effectiveness of the biological control of the Botrytis gray mould, caused by Botrytis cinerea Pers., one of the most important fungal diseases of the tomato (Lycopersicon esculentum Mill.). Biological control was performed by using Trichoderma harzianum Rifai, an antagonist that is a naturally occurring fungus found on some plants and in the soil worldwide. Trichoderma spp. are fungi diffused in nearly all agricultural soils and in other environments such as decaying wood. The object of this research is to find control strategies to reduce chemical treatments that cause damage to the environment and increase the pathogen resistance, applying the biological control by using T. harzianum against B. cinerea. A commercial product containing a natural isolate of T. harzianum is trichodex (Makhteshim Chemical Works, LTD). The research was performed in laboratory and in greenhouse. In laboratory, radial growth reduction of B. cinerea, in presence of T. harzianum, was calculated in relation to the growth of the pathogen control, by using a specific formula that measures the percentage of the inhibition of the radial mycelial growth. In greenhouse, starting from the tomato fruit setting, the research was carried out comparing, by a randomized complete block experiment design, replicated four times, the following treatments:1) untreated control; 2) pyrimethanil (400 g/L of a.i.), at 200 cc/hL of c.i. (pyrimidine fungicides); 3) trichodex at 100g/hL (1 kg/ha); 4) trichodex at 200 g/hL (2 kg/ha); 5) trichodex at 400 g/hL (4 kg/ha). Before fruit setting, the plots were all treated against Botrytis gray mould with iprodione 50% (100 g/hL), procymidone 50% (100 g/hL) and switch (Novartis plant protection) at 80 g/hL. In dual culture, the inhibition of B. cinerea radial mycelial growth was 76%. No inhibition halo was observed between B. cinerea and T. harzianum colonies but, after 3 days, the pathogen colony radius resulted no more than 1

Utp22p acts in concert with Utp8p to channel aminoacyl-tRNA from the nucleolus to the nuclear tRNA export receptor Los1p but not Msn5p.

PubMed

Eswara, Manoja B K; Clayton, Ashley; Mangroo, Dev

2012-12-01

Utp8p is an essential nucleolar protein that channels aminoacyl-tRNAs from aminoacyl-tRNA synthetases in the nucleolus to the nuclear tRNA export receptors located in the nucleoplasm and nuclear pore complex in Saccharomyces cerevisiae. Utp8p is also part of the U3 snoRNA-associated protein complex involved in 18S rRNA biogenesis in the nucleolus. We report that Utp22p, which is another member of the U3 snoRNA-associated protein complex, is also an intranuclear component of the nuclear tRNA export machinery. Depletion of Utp22p results in nuclear retention of mature tRNAs derived from intron-containing and intronless precursors. Moreover, Utp22p copurifies with the nuclear tRNA export receptor Los1p, the aminoacyl-tRNA synthetase Tys1p and Utp8p, but not with the RanGTPase Gsp1p and the nuclear tRNA export receptor Msn5p. Utp22p interacts directly with Utp8p and Los1p in a tRNA-independent manner in vitro. Utp22p also interacts directly with Tys1p, but this binding is stimulated when Tys1p is bound to tRNA. However, Utp22p, unlike Utp8p, does not bind tRNA saturably. These data suggest that Utp22p recruits Utp8p to aminoacyl-tRNA synthetases in the nucleolus to collect aminoacyl-tRNA and then accompanies the Utp8p-tRNA complex to deliver the aminoacyl-tRNAs to Los1p but not Msn5p. It is possible that Nrap/Nol6, the mammalian orthologue of Utp22p, plays a role in channelling aminoacyl-tRNA to the nuclear tRNA export receptor exportin-t.

Does co-inoculation of Lactuca serriola with endophytic and arbuscular mycorrhizal fungi improve plant growth in a polluted environment?

PubMed

Ważny, Rafał; Rozpądek, Piotr; Jędrzejczyk, Roman J; Śliwa, Marta; Stojakowska, Anna; Anielska, Teresa; Turnau, Katarzyna

2018-04-01

Phytoremediation of polluted sites can be improved by co-inoculation with mycorrhizal and endophytic fungi. In this study, the effects of single- and co-inoculation of Lactuca serriola with an arbuscular mycorrhizal (AM) fungus, Rhizoglomus intraradices, and endophytic fungi, Mucor sp. or Trichoderma asperellum, on plant growth, vitality, toxic metal accumulation, sesquiterpene lactone production and flavonoid concentration in the presence of toxic metals were evaluated. Inoculation with the AM fungus increased biomass yield of the plants grown on non-polluted and polluted substrate. Co-inoculation with the AM fungus and Mucor sp. resulted in increased biomass yield of plants cultivated on the polluted substrate, whereas co-inoculation with T. asperellum and the AM fungus increased plant biomass on the non-polluted substrate. In the presence of Mucor sp., mycorrhizal colonization and arbuscule richness were increased in the non-polluted substrate. Co-inoculation with the AM fungus and Mucor sp. increased Zn concentration in leaves and roots. The concentration of sesquiterpene lactones in plant leaves was decreased by AM fungus inoculation in both substrates. Despite enhanced host plant costs caused by maintaining symbiosis with numerous microorganisms, interaction of wild lettuce with both mycorrhizal and endophytic fungi was more beneficial than that with a single fungus. The study shows the potential of double inoculation in unfavourable environments, including agricultural areas and toxic metal-polluted areas.

Changes in ruminal bacterial community composition following feeding of alfalfa silage inoculated with a commercial silage inoculant

USDA-ARS?s Scientific Manuscript database

Some silage inoculants promote an increase in milk production, possibly through altering the rumen microflora. In this study, dairy cows fed alfalfa silage treated with the inoculant, Lactobacillus plantarum MTD/1 (LPS), were compared to cows fed untreated silage (Ctrl) with the objectives: 1) to de...

Effects of inoculation with organic-phosphorus-mineralizing bacteria on soybean (Glycine max) growth and indigenous bacterial community diversity.

PubMed

Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Li, Yang; Duan, Man-Li

2017-05-01

Three different organic-phosphorus-mineralizing bacteria (OPMB) strains were inoculated to soil planted with soybean (Glycine max), and their effects on soybean growth and indigenous bacterial community diversity were investigated. Inoculation with Pseudomonas fluorescens Z4-1 and Brevibacillus agri L7-1 increased organic phosphorus degradation by 22% and 30%, respectively, compared with the control at the mature stage. Strains P. fluorescens Z4-1 and B. agri L7-1 significantly improved the soil alkaline phosphatase activity, average well color development, and the soybean root activity. Terminal restriction fragment length polymorphism analysis demonstrated that P. fluorescens Z4-1 and B. agri L7-1 could persist in the soil at relative abundances of 2.0%-6.4% throughout soybean growth. Thus, P. fluorescens Z4-1 and B. agri L7-1 could potentially be used in organic-phosphorus-mineralizing biofertilizers. OPMB inoculation altered the genetic structure of the soil bacterial communities but had no apparent influence on the carbon source utilization profiles of the soil bacterial communities. Principal components analysis showed that the changes in the carbon source utilization profiles of bacterial community depended mainly on the plant growth stages rather than inoculation with OPMB. The results help to understand the evolution of the soil bacterial community after OPMB inoculation.

Dry Transfer Inoculation of Low-Moisture Spices Containing Antimicrobial Compounds.

PubMed

Hildebrandt, Ian M; Hu, Chuxuan; Grasso-Kelley, Elizabeth M; Ye, Peiran; Anderson, Nathan M; Keller, Susanne E

2017-02-01

Inoculation of a food product for use in subsequent validation studies typically makes use of a high concentration cocktail of microorganisms suspended in aqueous media. However, this inoculation method may prove difficult particularly when the food product is a low-moisture food containing antimicrobial compounds, such as some dried spices. In this study, a dry transfer method for inoculation of clove powder, oregano leaves, ginger powder, and ground black pepper with a five-serovar cocktail of Salmonella was developed and compared with a traditional aqueous inoculation procedure. Spices were inoculated at three levels, 10, 8, and 6 log CFU/g, by using both an aqueous suspension of Salmonella and a dry transfer of Salmonella from previously inoculated silica beads. At the highest inoculation level, the dry transfer method resulted in a significantly higher microbial load (P < 0.05) for ground cloves and oregano, but not for ginger and ground black pepper. At the intermediate inoculation level, differences were apparent only for ginger and black pepper. Inoculation levels of 6 log CFU/g resulted in recoveries below detection limits for both methods of inoculation. Additional examination on the survival of Salmonella on silica beads after inoculation and in clove powder after dry transfer from silica beads showed linear rates of decline, with a rate of -0.011 log CFU/g/day for beads and -0.015 log CFU/g/day for clove powder. The results suggest that dry transfer of Salmonella via inoculated silica beads is a viable alternative when traditional aqueous inoculation is not feasible.

Harzianolide, a novel plant growth regulator and systemic resistance elicitor from Trichoderma harzianum.

PubMed

Cai, Feng; Yu, Guanghui; Wang, Ping; Wei, Zhong; Fu, Lin; Shen, Qirong; Chen, Wei

2013-12-01

A detailed understanding of the effect of natural products on plant growth and protection will underpin new product development for plant production. The isolation and characterization of a known secondary metabolite named harzianolide from Trichoderma harzianum strain SQR-T037 were described, and the bioactivity of the purified compound as well as the crude metabolite extract in plant growth promotion and systemic resistance induction was investigated in this study. The results showed that harzianolide significantly promoted tomato seedling growth by up to 2.5-fold (dry weight) at a concentration of 0.1 ppm compared with the control. The result of root scan suggested that Trichoderma secondary metabolites may influence the early stages of plant growth through better root development for the enhancement of root length and tips. Both of the purified harzianolide and crude metabolite extract increased the activity of some defense-related enzymes to response to oxidative stress. Examination of six defense-related gene expression by real-time reverse transcription-PCR analysis revealed that harzianolide induces the expression of genes involved in the salicylic acid (PR1 and GLU) and jasmonate/ethylene (JERF3) signaling pathways while crude metabolite extract inhibited some gene expression (CHI-II and PGIP) related to basal defense in tomato plants. Further experiment showed that a subsequent challenge of harzianolide-pretreated plants with the pathogen Sclerotinia sclerotiorum resulted in higher systemic resistance by the reduction of lesion size. These results indicate that secondary metabolites of Trichoderma spp., like harzianolide, may play a novel role in both plant growth regulation and plant defense responses. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Inoculation message treatments for curbing noncommunicable disease development.

PubMed

Mason, Alicia M; Miller, Claude H

2013-07-01

To study the effect of various types of inoculation message treatments on resistance to persuasive and potentially deceptive health- and nutrition-related (HNR) content claims of commercial food advertisers. A three-phase experiment was conducted among 145 students from a Midwestern U.S. university. Quantitative statistical analyses were used to interpret the results. RESULTS provide clear evidence that integrating regulatory focus/fit considerations enhances the treatment effectiveness of inoculation messages. Inoculation messages that employed a preventative, outcome focus with concrete language were most effective at countering HNR advertising claims. The findings indicate that inoculation fosters resistance equally across the most common types of commercially advertised HNR product claims (e.g., absolute, general, and structure/function claims). As the drive to refine the inoculation process model continues, further testing and application of this strategy in a public health context is needed to counter ongoing efforts by commercial food advertisers to avoid government regulations against deceptive practices such as dubious health/nutrition claims. This research advances inoculation theory by providing evidence that 1) good regulatory fit strengthens the effect of refutational preemption and 2) an inoculation approach is highly effective at fostering resistance to commercial advertisers' HNR content claims. This macro approach appears far superior to education or information-based promotional health campaigns targeted solely at specific populations demonstrating rising rates of noncommunicable disease.

Klebsiella pneumoniae inoculants for enhancing plant growth

DOEpatents

Triplett, Eric W [Middleton, WI; Kaeppler, Shawn M [Oregon, WI; Chelius, Marisa K [Greeley, CO

2008-07-01

A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

A CD22-reactive TCR from the T-cell allorepertoire for the treatment of acute lymphoblastic leukemia by TCR gene transfer

PubMed Central

Jahn, Lorenz; Hagedoorn, Renate S.; van der Steen, Dirk M.; Hombrink, Pleun; Kester, Michel G.D.; Schoonakker, Marjolein P.; de Ridder, Daniëlle; van Veelen, Peter A.; Falkenburg, J.H. Frederik; Heemskerk, Mirjam H.M.

2016-01-01

CD22 is currently evaluated as a target-antigen for the treatment of B-cell malignancies using chimeric antigen receptor (CAR)-engineered T-cells or monoclonal antibodies (mAbs). CAR- and mAbs-based immunotherapies have been successfully applied targeting other antigens, however, occurrence of refractory disease to these interventions urges the identification of additional strategies. Here, we identified a TCR recognizing the CD22-derived peptide RPFPPHIQL (CD22RPF) presented in human leukocyte antigen (HLA)-B*07:02. To overcome tolerance to self-antigens such as CD22, we exploited the immunogenicity of allogeneic HLA. CD22RPF-specific T-cell clone 9D4 was isolated from a healthy HLA-B*07:02neg individual, efficiently produced cytokines upon stimulation with primary acute lymphoblastic leukemia and healthy B-cells, but did not react towards healthy hematopoietic and nonhematopoietic cell subsets, including dendritic cells (DCs) and macrophages expressing low levels of CD22. Gene transfer of TCR-9D4 installed potent CD22-specificity onto recipient CD8+ T-cells that recognized and lysed primary B-cell leukemia. TCR-transduced T-cells spared healthy CD22neg hematopoietic cell subsets but weakly lysed CD22low-expressing DCs and macrophages. CD22-specific TCR-engineered T-cells could form an additional immunotherapeutic strategy with a complementary role to CAR- and antibody-based interventions in the treatment of B-cell malignancies. However, CD22 expression on non-B-cells may limit the attractiveness of CD22 as target-antigen in cellular immunotherapy. PMID:27689397

A CD22-reactive TCR from the T-cell allorepertoire for the treatment of acute lymphoblastic leukemia by TCR gene transfer.

PubMed

Jahn, Lorenz; Hagedoorn, Renate S; van der Steen, Dirk M; Hombrink, Pleun; Kester, Michel G D; Schoonakker, Marjolein P; de Ridder, Daniëlle; van Veelen, Peter A; Falkenburg, J H Frederik; Heemskerk, Mirjam H M

2016-11-01

CD22 is currently evaluated as a target-antigen for the treatment of B-cell malignancies using chimeric antigen receptor (CAR)-engineered T-cells or monoclonal antibodies (mAbs). CAR- and mAbs-based immunotherapies have been successfully applied targeting other antigens, however, occurrence of refractory disease to these interventions urges the identification of additional strategies. Here, we identified a TCR recognizing the CD22-derived peptide RPFPPHIQL (CD22RPF) presented in human leukocyte antigen (HLA)-B*07:02. To overcome tolerance to self-antigens such as CD22, we exploited the immunogenicity of allogeneic HLA. CD22RPF-specific T-cell clone 9D4 was isolated from a healthy HLA-B*07:02neg individual, efficiently produced cytokines upon stimulation with primary acute lymphoblastic leukemia and healthy B-cells, but did not react towards healthy hematopoietic and nonhematopoietic cell subsets, including dendritic cells (DCs) and macrophages expressing low levels of CD22. Gene transfer of TCR-9D4 installed potent CD22-specificity onto recipient CD8+ T-cells that recognized and lysed primary B-cell leukemia. TCR-transduced T-cells spared healthy CD22neg hematopoietic cell subsets but weakly lysed CD22low-expressing DCs and macrophages. CD22-specific TCR-engineered T-cells could form an additional immunotherapeutic strategy with a complementary role to CAR- and antibody-based interventions in the treatment of B-cell malignancies. However, CD22 expression on non-B-cells may limit the attractiveness of CD22 as target-antigen in cellular immunotherapy.

Success rates of intrauterine inoculations of layers via the vagina.

PubMed

Landman, W J M; Matthijs, M G R; van Eck, J H H

2013-02-01

Intrauterine inoculation of layer hens has been documented previously in the literature; however, its efficiency has only been assessed on a very small scale. Attempts were therefore made to inoculate 14 experimental groups each consisting of 10 commercial table egg-producing hens intrauterinely with methylene blue. The effect of four variables-oviposition (natural, hormonally induced or not), position of the hen at inoculation (vertical/horizontal), technique to access the uterus (three methods) and inoculation device (four types)-on the success rate of intrauterine inoculation of layers of different breed and age was studied. Immediately after inoculation, hens were euthanized and the presence of methylene blue and perforations in the oviduct were examined. Successful inoculation rates ranged from 0/10 to 10/10. The 100% success rate was obtained in birds without an egg in the uterus, which were restrained in a horizontal position and inoculated with a hard catheter after exposure of the uterovaginal junction. The second best score was obtained in hens inoculated shortly after natural oviposition, maintained in a vertical position and using a corkscrew-shaped saliva ejector after exposure of the vaginal orifice (7/10). In all other groups the success rate was 5/10 or less. Vaginal perforation occurred in none to eight birds per group. It is concluded that intrauterine inoculation of laying hens is only reliable in birds without an egg in the uterus after exposure of the uterovaginal junction. All other methods proved unreliable and often caused vaginal perforations. However, intravaginal inoculation can be performed with 100% accuracy providing the vaginal orifice is exposed.

Effect of Microbial inoculation in combating the aluminium toxicity effect on growth of Zea mays.

PubMed

Arora, P; Singh, G; Tiwari, A

2017-07-31

The present study is aimed at improving the aluminium tolerance in maize crop employing the potential of microbial inoculants in conferring resistance to these toxicities via production of certain chelating compounds like siderophores, exopolysachharides and organic acids. Acid soils have now-a-days become one of the key factors for limiting growth of many agriculturally important crops. Aluminium  is one of the major elements present in acid soils and is mainly responsible for toxicity in the soil. This aluminium is rapidly soluble in soil water and hence absorbed by plant roots under conditions where soil pH is below 5. This toxicity leads to severe root growth inhibition, thereby limiting the production of maize crops. It was observed that use of microbial inoculums can be helpful in elimination of these toxic compounds and prevent the inhibition of root growth . It was found that the soils contaminated with aluminium toxicity decreased the root length of maize plant significantly by 65% but Bacillus and Burkholderia inoculation increased this root length significantly by 1.4- folds and 2- folds respectively thereby combating the effect of aluminium toxicity. Aluminium concentration was found maximum in roots of plants which were grown under aluminium stress condition. But this aluminium accumulation decreased ̴ 2-folds when Burkholderia was used as seed inoculants under aluminium stress conditions. Also, at 60mM aluminium accumulation, phosphorus solubilisation in roots was found to be increased upto 30% on Burkholderia inoculation. However, Bacillus inoculation didn't show any significant difference in either of the case. Thus, the inoculation of seeds with Burkholderia isolates could prove to be a boon in sequestering aluminium toxicity in Zea mays.

Automatic inoculating apparatus. [includes movable carraige, drive motor, and swabbing motor

NASA Technical Reports Server (NTRS)

Wilkins, J. R.; Mills, S. M. (Inventor)

1974-01-01

An automatic inoculating apparatus for agar trays is described and using a simple inoculating element, such as a cotton swab or inoculating loop. The apparatus includes a movable carriage for supporting the tray to be inoculated, a drive motor for moving the tray along a trackway, and a swabbing motor for automatically swabbing the tray during the movement. An actuator motor controls lowering of the inoculating element onto the tray and lifting of the inoculating element. An electrical control system, including limit microswitches, enables automatic control of the actuator motor and return of the carriage to the initial position after inoculating is completed.

Increased protein content of chickpea (Cicer arietinum L.) inoculated with arbuscular mycorrhizal fungi and nitrogen-fixing bacteria under water deficit conditions.

PubMed

Oliveira, Rui S; Carvalho, Patrícia; Marques, Guilhermina; Ferreira, Luís; Nunes, Mafalda; Rocha, Inês; Ma, Ying; Carvalho, Maria F; Vosátka, Miroslav; Freitas, Helena

2017-10-01

Chickpea (Cicer arietinum L.) is a widely cropped pulse and an important source of proteins for humans. In Mediterranean regions it is predicted that drought will reduce soil moisture and become a major issue in agricultural practice. Nitrogen (N)-fixing bacteria and arbuscular mycorrhizal (AM) fungi have the potential to improve plant growth and drought tolerance. The aim of the study was to assess the effects of N-fixing bacteria and AM fungi on the growth, grain yield and protein content of chickpea under water deficit. Plants inoculated with Mesorhizobium mediterraneum or Rhizophagus irregularis without water deficit and inoculated with M. mediterraneum under moderate water deficit had significant increases in biomass. Inoculation with microbial symbionts brought no benefits to chickpea under severe water deficit. However, under moderate water deficit grain crude protein was increased by 13%, 17% and 22% in plants inoculated with M. mediterraneum, R. irregularis and M. mediterraneum + R. irregularis, respectively. Inoculation with N-fixing bacteria and AM fungi has the potential to benefit agricultural production of chickpea under water deficit conditions and to contribute to increased grain protein content. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Differential display of abundantly expressed genes of Trichoderma harzianum during colonization of tomato-germinating seeds and roots.

PubMed

Mehrabi-Koushki, Mehdi; Rouhani, Hamid; Mahdikhani-Moghaddam, Esmat

2012-11-01

The identification of Trichoderma genes whose expression is altered during early stages of interaction with developing roots of germinated seeds is an important step toward understanding the rhizosphere competency of Trichoderma spp. The potential of 13 Trichoderma strains to colonize tomato root and promote plant growth has been evaluated. All used strains successfully propagated in spermosphere and continued their growth in rhizoplane simultaneously root enlargement while the strains T6 and T7 were the most abundant in the apical segment of roots. Root colonization in most strains associated with promoting the roots and shoots growth while they significantly increased up to 43 and 40 % roots and shoots dry weights, respectively. Differential display reverse transcriptase-PCR (DDRT-PCR) has been developed to detect differentially expressed genes in the previously selected strain, Trichoderma harzianum T7, during colonization stages of tomato-germinating seeds and roots. Amplified DDRT-PCR products were analyzed on gel agarose and 62 differential bands excised, purified, cloned, and sequenced. Obtained ESTs were submit-queried to NCBI database by BLASTx search and gene ontology hierarchy. Most of transcripts (29 EST) corresponds to known and hypothetical proteins such as secretion-related small GTPase, 40S ribosomal protein S3a, 3-hydroxybutyryl-CoA dehydrogenase, DNA repair protein rad50, lipid phosphate phosphatase-related protein type 3, nuclear essential protein, phospholipase A2, fatty acid desaturase, nuclear pore complex subunit Nup133, ubiquitin-activating enzyme, and 60S ribosomal protein L40. Also, 13 of these sequences showed no homology (E > 0.05) with public databases and considered as novel genes. Some of these ESTs corresponded to genes encodes enzymes potentially involved in nutritional support of microorganisms which have obvious importance in the establishment of Trichoderma in spermosphere and rhizosphere, via potentially functioning in

Molecular evolution and phylogenetic analysis of biocontrol genes acquired from SCoT polymorphism of mycoparasitic Trichoderma koningii inhibiting phytopathogen Rhizoctonia solani Kuhn.

PubMed

Gajera, H P; Hirpara, Darshna G; Katakpara, Zinkal A; Patel, S V; Golakiya, B A

2016-11-01

The biocontrol agent Trichoderma (T. harzianum, T. viride, T. virens, T. hamantum, T. koningii, T. pseudokoningii and Trichoderma species) inhibited variably (15.32 - 88.12%) the in vitro growth of Rhizoctonia solani causing root rot in cotton. The T. koningii MTCC 796 evidenced highest (88.12%) growth inhibition of test pathogen followed by T. viride NBAII Tv23 (85.34%). Scanning electron microscopic study confirmed mycoparasitism for MTCC 796 and Tv23 which were capable of completely overgrowing on R. solani by degrading mycelia, coiling around the hyphae with hook-like structures. The antagonists T. harzianum NBAII Th1 and, T. virens NBAII Tvs12 exhibited strong antibiosis and formed 2-4 mm zone of inhibition for 70.28% and 46.62%, respectively growth inhibition of test pathogen. Mycoparasitism is a strong mode of action for biocontrol activity compared with antibiosis. The antagonists Trichoderma strains were performed for start codon targeted (SCoT) polymorphism to acquire biocontrol genes from potent antagonist. The six unique SCoT fragments amplified by genomic DNA of best mycoparasitic antagonist MTCC 796 strain are subjected to DNA sequencing resulted to confirm two functional sequences for activity related to biocontrol genes. The phylogenetic and molecular evolution of functional 824 bp of SCoT-3 (920) and 776 bp of SCoT-6 (806) fragments signify sequence homology with biocontrol genes endochitinase (partial cds of 203 amino acids) and novel hmgR genes (partial cds of 239 amino acids), respectively and the same were annotated and deposited in NCBI GenBank database. The hmgR gene is liable to be express hmg - CoA reductase which is a key enzyme for regulation of terpene biosynthesis and mycoparasitic strains produced triterpenes during antagonism to inhibit growth of fungal pathogen as evidenced with GC-MS profile. The biocontrol genes are found in best antagonist T. koningii MTCC 796 for mycoparasitic activity to restrain the growth of test pathogen R

Differentiation of Trypanosoma brucei, T. rhodesiense, and T. gambiense by the indirect fluorescent antibody test

PubMed Central

Latif, B. M. A.; Adam, Katherine M. G.

1973-01-01

Epidemiological studies, if they are to lead to appropriate preventive procedures, require knowledge of the host distribution of the parasite. Progress in the epidemiology of African trypanosomiasis is restricted by the lack of a reliable and simple method of differentiating Trypanosoma brucei, T. rhodesiense, and T. gambiense. The recently introduced blood inoculation infectivity test promises to fulfil this need by distinguishing T. brucei from T. rhodesiense, but it would not be suitable for separating T. brucei from T. gambiense, since rats and mice are frequently refractory to infection by fresh isolates of T. gambiense. Previous studies had indicated that the indirect fluorescent antibody test might differentiate not only the subgenera of the salivarian trypanosome species but also members of the subgenus Trypanozoon. A method of performing the test is described that enables T. brucei, T. rhodesiense, and T. gambiense to be differentiated by the titre of the sera. The method might be used in conjunction with the blood inoculation infectivity test to distinguish between new isolates of the subgenus Trypanozoon in East Africa, and also to search for possible animal reservoirs of T. gambiense in West Africa. PMID:4587481

CD22 Promotes B-1b Cell Responses to T Cell-Independent Type 2 Antigens.

PubMed

Haas, Karen M; Johnson, Kristen L; Phipps, James P; Do, Cardinal

2018-03-01

CD22 (Siglec-2) is a critical regulator of B cell activation and survival. CD22 -/- mice generate significantly impaired Ab responses to T cell-independent type 2 (TI-2) Ags, including haptenated Ficoll and pneumococcal polysaccharides, Ags that elicit poor T cell help and activate BCR signaling via multivalent epitope crosslinking. This has been proposed to be due to impaired marginal zone (MZ) B cell development/maintenance in CD22 -/- mice. However, mice expressing a mutant form of CD22 unable to bind sialic acid ligands generated normal TI-2 Ab responses, despite significantly reduced MZ B cells. Moreover, mice treated with CD22 ligand-binding blocking mAbs, which deplete MZ B cells, had little effect on TI-2 Ab responses. We therefore investigated the effects of CD22 deficiency on B-1b cells, an innate-like B cell population that plays a key role in TI-2 Ab responses. B-1b cells from CD22 -/- mice had impaired BCR-induced proliferation and significantly increased intracellular Ca 2+ concentration responses following BCR crosslinking. Ag-specific B-1b cell expansion and plasmablast differentiation following TI-2 Ag immunization was significantly impaired in CD22 -/- mice, consistent with reduced TI-2 Ab responses. We generated CD22 -/- mice with reduced CD19 levels (CD22 -/- CD19 +/- ) to test the hypothesis that augmented B-1b cell BCR signaling in CD22 -/- mice contributes to impaired TI-2 Ab responses. BCR-induced proliferation and intracellular Ca 2+ concentration responses were normalized in CD22 -/- CD19 +/- B-1b cells. Consistent with this, TI-2 Ag-specific B-1b cell expansion, plasmablast differentiation, survival, and Ab responses were rescued in CD22 -/- CD19 +/- mice. Thus, CD22 plays a critical role in regulating TI-2 Ab responses through regulating B-1b cell signaling thresholds. Copyright © 2018 by The American Association of Immunologists, Inc.

Limited susceptibility of pigeons experimentally inoculated with H5N1 highly pathogenic avian influenza viruses.

PubMed

Yamamoto, Yu; Nakamura, Kikuyasu; Yamada, Manabu; Mase, Masaji

2012-02-01

An experimental infection study was performed using pigeons reared for racing or meat production in Japan and clade 2.2 and 2.3.2 isolates of H5N1 highly pathogenic avian influenza virus to evaluate the possible role of pigeons in virus transmission to poultry. In experiment 1, when 20 pigeons were intranasally inoculated with high or low viral doses, no inoculated pigeon exhibited clinical signs for 14 days. Drinking water and almost all swab samples were negative for virus isolation. Virus isolation was positive in 3 oral swab samples from 2 pigeons from day 2 through 4 postinoculation, but viral titers of positive samples were extremely low. Immunohistochemical analysis for virus detection was negative in all tissue samples. Along with seroconversion in a limited number of pigeons postinoculation, these results suggest that pigeons have limited susceptibility to the virus used for experimental infection. In experiment 2, when uninoculated chickens were housed with virus-inoculated pigeons, all pigeons and contact chickens survived for 14 days without exhibiting any clinical signs. According to serological analysis, the chickens did not exhibit seroconversion after close contact with inoculated pigeons. Our data suggest that the risk posed by pigeons with respect to the transmission of the H5N1 highly pathogenic avian influenza virus to poultry would be less than that for other susceptible avian species.

Solid-state fermentation of agro-industrial wastes to produce bioorganic fertilizer for the biocontrol of Fusarium wilt of cucumber in continuously cropped soil.

PubMed

Chen, Lihua; Yang, Xingming; Raza, Waseem; Luo, Jia; Zhang, Fengge; Shen, Qirong

2011-02-01

Agro-industrial wastes of cattle dung, vinegar-production residue and rice straw were solid-state fermented by inoculation with Trichoderma harzianum SQR-T037 (SQR-T037) for production of bioorganic fertilizers containing SQR-T037 and 6-pentyl-α-pyrone (6PAP) to control Fusarium wilt of cucumber in a continuously cropped soil. Fermentation days, temperature, inoculum and vinegar-production residue demonstrated significant effects on the SQR-T037 biomass and the yield of 6PAP, based on fractional factorial design. Three optimum conditions for producing the maximum SQR-T037 biomass and 6PAP yield were predicted by central composite design and validated. Bioorganic fertilizer containing 8.46 log(10) ITS copies g(-1) dry weight of SQR-T037 and 1291.73 mg kg(-1) dry weight of 6PAP, and having the highest (p<0.05) biocontrol efficacy, was achieved at 36.7 fermentation days, 25.9°C temperature, 7.6% inoculum content, 41.0% vinegar-production residue, 20.0% rice straw and 39.0% cattle dung. This is a way to offer a high value-added use for agro-industrial wastes. Copyright © 2010 Elsevier Ltd. All rights reserved.

The Combination of Trichoderma harzianum and Chemical Fertilization Leads to the Deregulation of Phytohormone Networking, Preventing the Adaptive Responses of Tomato Plants to Salt Stress.

PubMed

Rubio, M B; Hermosa, Rosa; Vicente, Rubén; Gómez-Acosta, Fabio A; Morcuende, Rosa; Monte, Enrique; Bettiol, Wagner

2017-01-01

Plants have evolved effective mechanisms to avoid or reduce the potential damage caused by abiotic stresses. In addition to biocontrol abilities, Trichoderma genus fungi promote growth and alleviate the adverse effects caused by saline stress in plants. Morphological, physiological, and molecular changes were analyzed in salt-stressed tomato plants grown under greenhouse conditions in order to investigate the effects of chemical and biological fertilizations. The application of Trichoderma harzianum T34 to tomato seeds had very positive effects on plant growth, independently of chemical fertilization. The application of salt stress significantly changed the parameters related to growth and gas-exchange rates in tomato plants subject to chemical fertilization. However, the gas-exchange parameters were not affected in unfertilized plants under the same moderate saline stress. The combined application of T34 and salt significantly reduced the fresh and dry weights of NPK-fertilized plants, while the opposite effects were detected when no chemical fertilization was applied. Decaying symptoms were observed in salt-stressed and chemically fertilized plants previously treated with T34. This damaged phenotype was linked to significantly higher intercellular CO 2 and slight increases in stomatal conductance and transpiration, and to the deregulation of phytohormone networking in terms of significantly lower expression levels of the salt overlay sensitivity 1 ( SOS1 ) gene, and the genes involved in signaling abscisic acid-, ethylene-, and salicylic acid-dependent pathways and ROS production, in comparison with those observed in salt-challenged NPK-fertilized plants.

Ionizing radiation and autoimmunity: Induction of autoimmune disease in mice by high dose fractionated total lymphoid irradiation and its prevention by inoculating normal T cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sakaguchi, N.; Sakaguchi, S.; Miyai, K.

1992-11-01

Ionizing radiation can functionally alter the immune system and break self-tolerance. High dose (42.5 Gy), fractionated (2.5 Gy 17 times) total lymphoid irradiation (TLI) on mice caused various organ-specific autoimmune diseases, such as gastritis, thyroiditis, and orchitis, depending on the radiation dosages, the extent of lymphoid irradiation, and the genetic background of the mouse strains. Radiation-induced tissue damage is not the primary cause of the autoimmune disease because irradiation of the target organs alone failed to elicit the autoimmunity and shielding of the organs from irradiation was unable to prevent it. In contrast, irradiation of both the thymus and themore » peripheral lymphoid organs/tissues was required for efficient induction of autoimmune disease by TLI. TLI eliminated the majority of mature thymocytes and the peripheral T cells for 1 mo, and inoculation of spleen cell, thymocyte, or bone marrow cell suspensions (prepared from syngeneic nonirradiated mice) within 2 wk after TLI effectively prevented the autoimmune development. Depletion of T cells from the inocula abrogated the preventive activity. CD4[sup +] T cells mediated the autoimmune prevention but CD8[sup +] T cells did not. CD4[sup +] T cells also appeared to mediate the TLI-induced autoimmune disease because CD4[sup +] T cells from disease-bearing TLI mice adoptively transferred the autoimmune disease to syngeneic naive mice. Taken together, these results indicate that high dose, fractionated ionizing radiation on the lymphoid organs/tissues can cause autoimmune disease by affecting the T cell immune system, rather than the target self-Ags, presumably by altering T cell-dependent control of self-reactive T cells. 62 refs., 9 figs., 2 tabs.« less

Developing microbial inoculants for native Hawaiian trees

Treesearch

Kim H. Wilkinson

2002-01-01

We use certain microbial inoculants in the nursery as a part of supporting the objectives of accelerating rehabilitation of degraded land and ecosystem function, as well as reducing costs in establishment and maintenance of forest plantings. Microbial inoculants re-create natural partnerships between plants and some of the beneficial microorganisms that support plants...

Observations on the removal of brood inoculated with Tropilaelaps mercedesae (Mesostigmata: Laelapidae) and the mite’s reproductive success in Apis mellifera colonies

USDA-ARS?s Scientific Manuscript database

This study assessed the response of Apis mellifera to brood deliberately infested with Tropilaelaps mercedesae. The reproductive success of T. mercedesae in mite-inoculated and naturally infested brood was also compared. The presence of T. mercedesae inside brood cells significantly affected brood ...

Improvement of hydrogen fermentation of galactose by combined inoculation strategy.

PubMed

Sivagurunathan, Periyasamy; Anburajan, Parthiban; Kumar, Gopalakrishnan; Arivalagan, Pugazhendhi; Bakonyi, Péter; Kim, Sang-Hyoun

2017-03-01

This study evaluated the feasibility of anaerobic hydrogen fermentation of galactose, a red algal biomass sugar, using individual and combined mixed culture inocula. Heat-treated (90°C, 30 min) samples of granular sludge (GS) and suspended digester sludge (SDS) were used as inoculum sources. The type of mixed culture inoculum played an important role in hydrogen production from galactose. Between two inocula, granular sludge showed higher hydrogen production rate (HPR) and hydrogen yield (HY) of 2.2 L H 2 /L-d and 1.09 mol H 2 /mol galactose added , respectively. Combined inoculation (GS + SDS) led to an elevated HPR and HY of 3.1 L H 2 /L-d and 1.28 mol H 2 /mol galactose added , respectively. Acetic and butyric acids are the major organic acids during fermentation. Quantitative polymerase chain reaction (qPCR) revealed that the mixed culture generated using the combined inoculation contained a higher cluster I Clostridium abundance than the culture produced using the single inoculum. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Inducing Resistance to Conspiracy Theory Propaganda: Testing Inoculation and Metainoculation Strategies

ERIC Educational Resources Information Center

Banas, John A.; Miller, Gregory

2013-01-01

This investigation examined the boundaries of inoculation theory by examining how inoculation can be applied to conspiracy theory propaganda as well as inoculation itself (called metainoculation). A 3-phase experiment with 312 participants compared 3 main groups: no-treatment control, inoculation, and metainoculation. Research questions explored…

Evolution of infection in mice inoculated by the oral route with different developmental forms of Trypanosoma cruzi I and II.

PubMed

Dias, Greicy Brisa Malaquias; Gruendling, Ana Paula; Araújo, Silvana Marques; Gomes, Mônica Lúcia; Toledo, Max Jean de Ornelas

2013-11-01

Oral infection has become the most important transmission mechanism of Chagas disease in Brazil. For this study, the development of Trypanosoma cruzi infection in mice, induced by the oral and intraperitoneal (IP) routes, was compared. Four groups of Swiss mice were used to evaluate the influence of parasite genetics, number of parasites, inoculation volume and developmental stages on the development of the orally induced infection: 1 - blood trypomastigotes (BT) via oral; 2 - BT via IP; 3 - culture metacyclic trypomastigotes (MT) via oral; and 4 - culture MT via IP. Animals inoculated orally showed levels of parasitemia, as well as infectivity and mortality rates, lower than animals inoculated via IP, regardless of DTU (discrete typing unit) and inoculum. Animals infected with TcII showed higher levels of these parameters than did animals infected with TcI. The larger volume of inoculum showed a greater capacity to cause an infection when administered via the oral route. BT infection was more virulent than culture MT infection for both routes (oral and IP). However, mice inoculated orally with BT showed lower levels than via IP, while mice inoculated orally with culture MT showed similar levels of infection to those inoculated via IP. Mice inoculated with culture MT showed more histopathological changes than those inoculated with BT, regardless of the inoculation route. These results indicate that this alternative experimental model is useful for evaluating infection by T. cruzi isolates with subpatent parasitemia and low virulence, such as those belonging to the TcI and TcIV DTUs, which are prevalent in outbreaks of orally transmitted Chagas disease. Copyright © 2013 Elsevier Inc. All rights reserved.

Human herpes virus type 6 can cause skin lesions at the BCG inoculation site similar to Kawasaki Disease.

PubMed

Kakisaka, Yosuke; Ohara, Tomoichiro; Katayama, Saori; Suzuki, Tasuku; Sasai, Shu; Hino-Fukuyo, Naomi; Kure, Shigeo

2012-12-01

Kawasaki Disease (KD) is acute, febrile, multisystem vasculitis of early childhood, the detailed mechanism of which is still unclear. Skin symptoms occur in KD, such as edema of the hands and feet with subsequent desquamation and redness at the inoculation site of bacillus Calmette-Guerin (BCG). The change at the BCG inoculation site has been considered as a specific feature of KD, although its mechanism is not fully understood. We present an 11-month-old boy who developed fever with redness of the BCG site due to infection with human herpes virus type 6 (HHV6). At the age of 3 months, the patient received BCG. His fever remitted 7 days after the onset of skin redness, with sequential desquamation at the BCG site and extremities, which is not a common feature of HHV6 infection that typically lasts for 3 days. The final diagnosis was exanthema subitum. Characteristically, the HHV6 infection in our patient appeared to be associated with the invigoration of the T cell system, as represented by the elevated serum levels of soluble interleukin-2 receptor (3,490 U/ml vs. normal range 145-519 U/ml). This patient clearly showed redness and crusting at the BCG inoculation site, suggesting that HHV6 infection might cause skin changes similar to those of KD via an unknown mechanism. In addition, we suggest that the activation of the T cell system may account for the skin lesions in KD, characterized by redness and subsequent crusting of the BCG inoculation site and desquamation of the extremities.

Production of bioethanol by direct bioconversion of oil-palm industrial effluent in a stirred-tank bioreactor.

PubMed

Alam, Md Zahangir; Kabbashi, Nassereldeen A; Hussin, S Nahdatul I S

2009-06-01

The purpose of this study was to evaluate the feasibility of producing bioethanol from palm-oil mill effluent generated by the oil-palm industries through direct bioconversion process. The bioethanol production was carried out through the treatment of compatible mixed cultures such as Thrichoderma harzianum, Phanerochaete chrysosporium, Mucor hiemalis, and yeast, Saccharomyces cerevisiae. Simultaneous inoculation of T. harzianum and S. cerevisiae was found to be the mixed culture that yielded the highest ethanol production (4% v/v or 31.6 g/l). Statistical optimization was carried out to determine the operating conditions of the stirred-tank bioreactor for maximum bioethanol production by a two-level fractional factorial design with a single central point. The factors involved were oxygen saturation level (pO(2)%), temperature, and pH. A polynomial regression model was developed using the experimental data including the linear, quadratic, and interaction effects. Statistical analysis showed that the maximum ethanol production of 4.6% (v/v) or 36.3 g/l was achieved at a temperature of 32 degrees C, pH of 6, and pO(2) of 30%. The results of the model validation test under the developed optimum process conditions indicated that the maximum production was increased from 4.6% (v/v) to 6.5% (v/v) or 51.3 g/l with 89.1% chemical-oxygen-demand removal.

Cancer cell-derived IL-1α induces CCL22 and the recruitment of regulatory T cells

PubMed Central

Wiedemann, Gabriela Maria; Knott, Max Martin Ludwig; Vetter, Viola Katharina; Rapp, Moritz; Haubner, Sascha; Fesseler, Julia; Kühnemuth, Benjamin; Layritz, Patrick; Thaler, Raffael; Kruger, Stephan; Ormanns, Steffen; Mayr, Doris; Endres, Stefan; Anz, David

2016-01-01

ABSTRACT In cancer patients, immunosuppression through regulatory T cells (Treg) is a crucial component of tumor immune evasion and contributes to disease progression. Tumor-infiltrating Treg in particular suppress local effector T cell responses and are associated with poor prognosis in tumors such as human pancreatic cancer or hepatocellular carcinoma (HCC). The chemokine CCL22 is known to recruit Treg into the tumor tissue and many types of human tumors are known to express high levels of CCL22. The mechanisms leading to intratumoral secretion of CCL22 are so far unknown. We demonstrate here that intratumoral CCL22 is induced in tumor-infiltrating immune cells through cancer cell-derived interleukin-1 (IL-1α). In pancreatic cancer and HCC, CCL22 is produced by intratumoral dendritic cells, while the cancer cells themselves do not secrete CCL22 in vitro and in vivo. Incubation of human peripheral blood mononuclear cells (PBMC) or murine splenocytes with tumor cells or tumor cell supernatants strongly induced CCL22 secretion in vitro. Tumor cell supernatants contained IL-1 and CCL22 induction in PBMC could be specifically prevented by the IL-1 receptor antagonist anakinra or by transfection of tumor cell lines with IL-1 siRNA, leading to a suppression of Treg migration. In conclusion, we identify here tumor cell-derived IL-1α as a major inducer of the Treg attracting chemokine CCL22 in human cancer cells. Therapeutic blockade of the IL-1 pathway could represent a promising strategy to inhibit tumor-induced immunosuppression. PMID:27757295

Changes in selected enzyme activities during growth of pure and mixed cultures of the white-rot decay fungus Trametes versicolor and the potential biocontrol fungus Trichoderma harzianum.

PubMed

Freitag, M; Morrell, J J

1992-04-01

Two filamentous fungi, the white-rot fungus Trametes versicolor and the soil fungus and potential biocontrol organism Trichoderma harzianum, have been grown in pure and mixed cultures on low-N (0.4 mM) and high-N (4 mM) defined synthetic media to determine the activities of selected wood-degrading enzymes such as cellobiase, cellulase, laccase, and peroxidases. Growth characteristics and enzyme activities were examined for potential correlations. Such correlations would allow the use of simple enzyme assays for measuring biomass development and would facilitate predictions about competitiveness of species in mixed fungal cultures. Our results show that while laccase and Poly Red-478 peroxidase activities indicate survival of the decay fungus, none of the monitored extracellular enzymes can serve as a quantitative indicator for biomass accumulation. As expected, the level of available nitrogen affected the production of the enzymes monitored: in low-N media, specific cellobiase, specific cellulase, and peroxidase activities were enhanced, while laccase activities were reduced. Most importantly, laccase activities of Trametes versicolor, and to a smaller extent, cellobiase activities of both fungi, were significantly induced in mixed cultures of Trametes versicolor and Trichoderma harzianum.

Application of gamma irradiation for inactivation of three pathogenic bacteria inoculated into meatballs

NASA Astrophysics Data System (ADS)

Gumus, Tuncay; Şukru Demirci, A.; Murat Velioglu, H.; Velioglu, Serap D.; Yilmaz, Ismail; Sagdic, Osman

2008-09-01

In this research, the effect of gamma irradiation on the inactivation of Escherichia coli O157:H7 (ATCC 33150), Staphylococcus aureus (ATCC 2392) and Salmonella typhimurium (NRRL 4463) inoculated into Tekirdag meatballs was investigated. The meatball samples were inoculated with pathogens and irradiated at the absorbed doses of 1, 2.2, 3.2, 4.5 and 5.2 kGy. E. coli O157:H7 count in 1 kGy irradiated meatballs stored in the refrigerator for 7 days was detected to be 4 log cfu/g lower than the count in nonirradiated samples ( p<0.05). S. aureus counts were decreased to 4 log cfu/g after being exposed to irradiation at a dose of 1 kGy. Although it was ineffective on elimination of S. typhimurium, irradiation at a dose of 3.2 kGy reduced E. coli O157:H7 and S. aureus counts under detectable values in the meatballs. However, none of the test organisms were detected in the samples after irradiation with 4.5 kGy doses.

Inoculation effect in prevention of increased verbal aggression in schools.

PubMed

Rosenberg, Steven

2004-12-01

This paper presents an argument for the use of intervention programs as inoculation agents in adolescents' social behavior, specifically the prevention of abusive and antisocial verbal aggression. The theoretical substance of the intervention is social-cognitive and eclectic, borrowing from several strategies known to be effective. The bridge between theory and curriculum is an inoculation-like process. Inoculation is not a strategy to change beliefs but rather to reinforce prosocial attitudes and assist students in overcoming influences that might lessen their prosocial stance. Results must include a control group to assess the effect of inoculation.

Efficacy of different fungicides against Rhizoctonia brown patch and Pythium blight on turfgrass in Italy.

PubMed

Mocioni, M; Titone, P; Garibaldi, A; Gullino, M L

2003-01-01

Brown patch, incited by Rhizoctonia solani Kuhn, and Pythium blight, caused by Pythium spp. are two of the diseases most frequently observed on turfgrass in high maintenance stands, as on golf courses. In such conditions the control strategies, based on chemicals, are particularly difficult due to the scarcity of fungicides registered for turf in Italy. The results obtained in experimental trials carried out to evaluate the efficacy of chemical and biological products against brown patch and Pythium blight are reported. On mature turfgrass, maintained under fairway conditions, azoxystrobin, and trifoxystrobin, not yet registered on turf, were very effective against brown patch. Tebuconazole, applied in three different formulations, was very effective against R. solani, while Trichoderma spp. and azadiractine did not control the pathogen. In greenhouse conditions on Agrostis stolonifera, in the presence of severe disease incidence, due to artificial inoculation, benalaxyl-M satisfactorily controlled Pythium blight; Trichoderma spp. as well as a commercial formulation of T. harzianum, applied one week before the inoculation, were not effective. Among the fungicides not yet registered for use on turfgrass in Italy, metalaxyl-M + mancozeb was effective against Pythium blight.

Short- and Long-Term Clinical Outcomes in Rhesus Monkeys Inoculated with a Highly Pathogenic Chimeric Simian/Human Immunodeficiency Virus

PubMed Central

Endo, Yasuyuki; Igarashi, Tatsuhiko; Nishimura, Yoshiaki; Buckler, Charles; Buckler-White, Alicia; Plishka, Ronald; Dimitrov, Dimiter S.; Martin, Malcolm A.

2000-01-01

A highly pathogenic simian/human immunodeficiency virus (SHIV), SHIVDH12R, isolated from a rhesus macaque that had been treated with anti-human CD8 monoclonal antibody at the time of primary infection with the nonpathogenic, molecularly cloned SHIVDH12, induced marked and rapid CD4+ T cell loss in all rhesus macaques intravenously inoculated with 1.0 50% tissue culture infective dose (TCID50) to 4.1 × 105 TCID50s of virus. Animals inoculated with 650 TCID50s of SHIVDH12R or more experienced irreversible CD4+ T lymphocyte depletion and developed clinical disease requiring euthanasia between weeks 12 and 23 postinfection. In contrast, the CD4+ T-cell numbers in four of five monkeys receiving 25 TCID50s of SHIVDH12R or less stabilized at low levels, and these surviving animals produced antibodies capable of neutralizing SHIVDH12R. In the fifth monkey, no recovery from the CD4+ T cell decline occurred, and the animal had to be euthanized. Viral RNA levels, subsequent to the initial peak of infection but not at peak viremia, correlated with the virus inoculum size and the eventual clinical course. Both initial infection rate constants, k, and decay constants, d, were determined, but only the latter were statistically correlated to clinical outcome. The attenuating effects of reduced inoculum size were also observed when virus was inoculated by the mucosal route. Because the uncloned SHIVDH12R stock possessed the genetic properties of a lentivirus quasispecies, we were able to assess the evolution of the input virus swarm in animals surviving the acute infection by monitoring the emergence of neutralization escape viral variants. PMID:10888632

Effect of High Dietary Manganese on the Immune Responses of Broilers Following Oral Salmonella typhimurium Inoculation.

PubMed

Pan, Shuqin; Zhang, Keying; Ding, Xuemei; Wang, Jianping; Peng, Huanwei; Zeng, Qiufeng; Xuan, Yue; Su, Zuowei; Wu, Bing; Bai, Shiping

2018-02-01

Manganese (Mn) is an essential nutrient for both host and pathogen. Recent studies have demonstrated the nutritional immunity of Mn against Salmonella infection in mammals. To investigate the effect of high dietary Mn on immune responses of broilers following Salmonella challenge, 144 1-day-old male broilers were fed a basal diet (containing 20.04 mg Mn/kg) plus an additional 40 (the control group) or 400 mg Mn/kg (the H-Mn group) for 7 days. The 72 broilers in each group were then orally inoculated with 5 × 10 7 CFUs of Salmonella typhimurium (ATCC#14028) or phosphate-buffered saline. Peripheral blood, spleens, cecal tonsils, and bursa of Fabricius were collected from Salmonella-inoculated and Salmonella-noninoculated broilers (n = 6) at 2 days post inoculation (2 DPI) and 7 days post inoculation (7 DPI). Peripheral blood lymphocyte subpopulations were determined by flow cytometry. The messenger RNA (mRNA) abundance of genes was determined by quantitative real-time polymerase chain reaction. Salmonella counts were higher (P < 0.05) in the H-Mn group than that in the control group at 2 DPI in the cecal contents of Salmonella-inoculated broilers. High dietary Mn increased CD3 + CD4 + and CD3 + CD8 + percentages in the peripheral blood of Salmonella-inoculated broilers at 2 DPI. Salmonella inoculation increased interleukin (IL)-6 mRNA expression in spleens and bursa of Fabricius at 2 DPI and increased IL-1β and IL-6 mRNA expression in cecal tonsils at 7 DPI in the H-Mn group. These changes were not observed in the control group. High dietary Mn increased interferon-γ (IFN-γ) in spleens and decreased IFN-γ and IL-12 mRNA expression in cecal tonsils of Salmonella-inoculated broilers at 2 DPI. High dietary Mn decreased IL-17 mRNA expression in the bursa of Fabricius at 7 DPI, but increased this expression in cecal tonsils at 2 and 7 DPI in Salmonella-inoculated broilers. These results suggested that dietary Mn level affected T helper (Th) 1-cytokine

Biogenic silver nanoparticles based on trichoderma harzianum: synthesis, characterization, toxicity evaluation and biological activity

NASA Astrophysics Data System (ADS)

Guilger, Mariana; Pasquoto-Stigliani, Tatiane; Bilesky-Jose, Natália; Grillo, Renato; Abhilash, P. C.; Fraceto, Leonardo Fernandes; Lima, Renata De

2017-03-01

White mold is an agricultural disease caused by the fungus Sclerotinia sclerotiorum, which affects important crops. There are different ways of controlling this organism, but none provides inhibition of its resistance structures (sclerotia). Nanotechnology offers promising applications in agricultural area. Here, silver nanoparticles were biogenically synthesized using the fungus Trichoderma harzianum and characterized. Cytotoxicity and genotoxicity were evaluated, and the nanoparticles were initially tested against white mold sclerotia. Their effects on soybean were also investigated with no effects observed. The nanoparticles showed potential against S. sclerotiorum, inhibiting sclerotia germination and mycelial growth. Nanoparticle characterization data indicated spherical morphology, satisfactory polydispersity and size distribution. Cytotoxicity and genotoxicity assays showed that the nanoparticles caused both the effects, although, the most toxic concentrations were above those applied for white mold control. Given the potential of the nanoparticles against S. sclerotiorum, we conclude that this study presents a first step for a new alternative in white mold control.

Intracellular CXCR4+ cell targeting with T22-empowered protein-only nanoparticles

PubMed Central

Unzueta, Ugutz; Céspedes, María Virtudes; Ferrer-Miralles, Neus; Casanova, Isolda; Cedano, Juan; Corchero, José Luis; Domingo-Espín, Joan; Villaverde, Antonio; Mangues, Ramón; Vázquez, Esther

2012-01-01

Background Cell-targeting peptides or proteins are appealing tools in nanomedicine and innovative medicines because they increase the local drug concentration and reduce potential side effects. CXC chemokine receptor 4 (CXCR4) is a cell surface marker associated with several severe human pathologies, including colorectal cancer, for which intracellular targeting agents are currently missing. Results Four different peptides that bind CXCR4 were tested for their ability to internalize a green fluorescent protein-based reporter nanoparticle into CXCR4+ cells. Among them, only the 18 mer peptide T22, an engineered segment derivative of polyphemusin II from the horseshoe crab, efficiently penetrated target cells via a rapid, receptor-specific endosomal route. This resulted in accumulation of the reporter nanoparticle in a fully fluorescent and stable form in the perinuclear region of the target cells, without toxicity either in cell culture or in an in vivo model of metastatic colorectal cancer. Conclusion Given the urgent demand for targeting agents in the research, diagnosis, and treatment of CXCR4-linked diseases, including colorectal cancer and human immunodeficiency virus infection, T22 appears to be a promising tag for the intracellular delivery of protein drugs, nanoparticles, and imaging agents. PMID:22923991

Nitrogen Metabolism and Growth Enhancement in Tomato Plants Challenged with Trichoderma harzianum Expressing the Aspergillus nidulans Acetamidase amdS Gene

PubMed Central

Domínguez, Sara; Rubio, M. Belén; Cardoza, Rosa E.; Gutiérrez, Santiago; Nicolás, Carlos; Bettiol, Wagner; Hermosa, Rosa; Monte, Enrique

2016-01-01

Trichoderma is a fungal genus that includes species that are currently being used as biological control agents and/or as biofertilizers. In addition to the direct application of Trichoderma spp. as biocontrol agents in plant protection, recent studies have focused on the beneficial responses exerted on plants, stimulating the growth, activating the defenses, and/or improving nutrient uptake. The amdS gene, encoding an acetamidase of Aspergillus, has been used as a selectable marker for the transformation of filamentous fungi, including Trichoderma spp., but the physiological effects of the introduction of this gene into the genome of these microorganisms still remains unexplored. No evidence of amdS orthologous genes has been detected within the Trichoderma spp. genomes and the amdS heterologous expression in Trichoderma harzianum T34 did not affect the growth of this fungus in media lacking acetamide. However, it did confer the ability for the fungus to use this amide as a nitrogen source. Although a similar antagonistic behavior was observed for T34 and amdS transformants in dual cultures against Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum, a significantly higher antifungal activity was detected in amdS transformants against F. oxysporum, compared to that of T34, in membrane assays on media lacking acetamide. In Trichoderma-tomato interaction assays, amdS transformants were able to promote plant growth to a greater extent than the wild-type T34, although compared with this strain the transformants showed similar capability to colonize tomato roots. Gene expression patterns from aerial parts of 3-week-old tomato plants treated with T34 and the amdS transformants have also been investigated using GeneChip Tomato Genome Arrays. The downregulation of defense genes and the upregulation of carbon and nitrogen metabolism genes observed in the microarrays were accompanied by (i) enhanced growth, (ii) increased carbon and nitrogen levels, and (iii) a

Nitrogen Metabolism and Growth Enhancement in Tomato Plants Challenged with Trichoderma harzianum Expressing the Aspergillus nidulans Acetamidase amdS Gene.

PubMed

Domínguez, Sara; Rubio, M Belén; Cardoza, Rosa E; Gutiérrez, Santiago; Nicolás, Carlos; Bettiol, Wagner; Hermosa, Rosa; Monte, Enrique

2016-01-01

Trichoderma is a fungal genus that includes species that are currently being used as biological control agents and/or as biofertilizers. In addition to the direct application of Trichoderma spp. as biocontrol agents in plant protection, recent studies have focused on the beneficial responses exerted on plants, stimulating the growth, activating the defenses, and/or improving nutrient uptake. The amdS gene, encoding an acetamidase of Aspergillus, has been used as a selectable marker for the transformation of filamentous fungi, including Trichoderma spp., but the physiological effects of the introduction of this gene into the genome of these microorganisms still remains unexplored. No evidence of amdS orthologous genes has been detected within the Trichoderma spp. genomes and the amdS heterologous expression in Trichoderma harzianum T34 did not affect the growth of this fungus in media lacking acetamide. However, it did confer the ability for the fungus to use this amide as a nitrogen source. Although a similar antagonistic behavior was observed for T34 and amdS transformants in dual cultures against Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum, a significantly higher antifungal activity was detected in amdS transformants against F. oxysporum, compared to that of T34, in membrane assays on media lacking acetamide. In Trichoderma-tomato interaction assays, amdS transformants were able to promote plant growth to a greater extent than the wild-type T34, although compared with this strain the transformants showed similar capability to colonize tomato roots. Gene expression patterns from aerial parts of 3-week-old tomato plants treated with T34 and the amdS transformants have also been investigated using GeneChip Tomato Genome Arrays. The downregulation of defense genes and the upregulation of carbon and nitrogen metabolism genes observed in the microarrays were accompanied by (i) enhanced growth, (ii) increased carbon and nitrogen levels, and (iii) a

Atmospheric pressure plasma jet treatment of Salmonella Enteritidis inoculated eggshells.

PubMed

Moritz, Maike; Wiacek, Claudia; Koethe, Martin; Braun, Peggy G

2017-03-20

Contamination of eggshells with Salmonella Enteritidis remains a food safety concern. In many cases human salmonellosis within the EU can be traced back to raw or undercooked eggs and egg products. Atmospheric pressure plasma is a novel decontamination method that can reduce a wide range of pathogens. The aim of this work was to evaluate the possibility of using an effective short time cold plasma treatment to inactivate Salmonella Enteritidis on the eggshell. Therefore, artificially contaminated eggshells were treated with an atmospheric pressure plasma jet under different experimental settings with various exposure times (15-300s), distances from the plasma jet nozzle to the eggshell surface (5, 8 or 12mm), feed gas compositions (Ar, Ar with 0.2, 0.5 or 1.0% O 2 ), gas flow rates (5 and 7slm) and different inoculations of Salmonella Enteritidis (10 1 -10 6 CFU/cm 2 ). Atmospheric pressure plasma could reduce Salmonella Enteritidis on eggshells significantly. Reduction factors ranged between 0.22 and 2.27 log CFU (colony-forming units). Exposure time and, particularly at 10 4 CFU/cm 2 inoculation, feed gas had a major impact on Salmonella reduction. Precisely, longer exposure times led to higher reductions and Ar as feed gas was more effective than ArO 2 mixtures. Copyright © 2017 Elsevier B.V. All rights reserved.

Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species

PubMed Central

Chen, Jing; Mao, Li-Juan; Feng, Xiao-Xiao; Zhang, Chu-Long; Lin, Fu-Cheng

2016-01-01

We surveyed the Trichoderma (Hypocreales, Ascomycota) biodiversity in agricultural fields in four major agricultural provinces of East China. Trichoderma strains were identified based on molecular approaches and morphological characteristics. In three sampled seasons (spring, summer and autumn), 2078 strains were isolated and identified to 17 known species: T. harzianum (429 isolates), T. asperellum (425), T. hamatum (397), T. virens (340), T. koningiopsis (248), T. brevicompactum (73), T. atroviride (73), T. fertile (26), T. longibrachiatum (22), T. pleuroticola (16), T. erinaceum (16), T. oblongisporum (2), T. polysporum (2), T. spirale (2), T. capillare (2), T. velutinum (2), and T. saturnisporum (1). T. harzianum, T. asperellum, T. hamatum, and T. virens were identified as the dominant species with dominance (Y) values of 0.057, 0.052, 0.048, and 0.039, respectively. The species amount, isolate numbers and the dominant species of Trichoderma varied between provinces. Zhejiang Province has shown the highest diversity, which was reflected in the highest species amount (14) and the highest Shannon–Wiener diversity index of Trichoderma haplotypes (1.46). We observed that relative frequencies of T. hamatum and T. koningiopsis under rice soil were higher than those under wheat and maize soil, indicating the preference of Trichoderma to different crops. Remarkable seasonal variation was shown, with summer exhibiting the highest biodiversity of the studied seasons. These results show that Trichoderma biodiversity in agricultural fields varies by region, crop, and season. Zhejiang Province (the southernmost province in the investigated area) had more T. hamatum than Shandong Province (the northernmost province), not only in isolate amounts but also in haplotype amounts. Furthermore, at haplotype level, only T. hamatum showed a gradient distribution from south to north in correspondence analysis among the four dominant species. The above results would contribute to the

Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species.

PubMed

Jiang, Yuan; Wang, Jin-Liang; Chen, Jing; Mao, Li-Juan; Feng, Xiao-Xiao; Zhang, Chu-Long; Lin, Fu-Cheng

2016-01-01

We surveyed the Trichoderma (Hypocreales, Ascomycota) biodiversity in agricultural fields in four major agricultural provinces of East China. Trichoderma strains were identified based on molecular approaches and morphological characteristics. In three sampled seasons (spring, summer and autumn), 2078 strains were isolated and identified to 17 known species: T. harzianum (429 isolates), T. asperellum (425), T. hamatum (397), T. virens (340), T. koningiopsis (248), T. brevicompactum (73), T. atroviride (73), T. fertile (26), T. longibrachiatum (22), T. pleuroticola (16), T. erinaceum (16), T. oblongisporum (2), T. polysporum (2), T. spirale (2), T. capillare (2), T. velutinum (2), and T. saturnisporum (1). T. harzianum, T. asperellum, T. hamatum, and T. virens were identified as the dominant species with dominance (Y) values of 0.057, 0.052, 0.048, and 0.039, respectively. The species amount, isolate numbers and the dominant species of Trichoderma varied between provinces. Zhejiang Province has shown the highest diversity, which was reflected in the highest species amount (14) and the highest Shannon-Wiener diversity index of Trichoderma haplotypes (1.46). We observed that relative frequencies of T. hamatum and T. koningiopsis under rice soil were higher than those under wheat and maize soil, indicating the preference of Trichoderma to different crops. Remarkable seasonal variation was shown, with summer exhibiting the highest biodiversity of the studied seasons. These results show that Trichoderma biodiversity in agricultural fields varies by region, crop, and season. Zhejiang Province (the southernmost province in the investigated area) had more T. hamatum than Shandong Province (the northernmost province), not only in isolate amounts but also in haplotype amounts. Furthermore, at haplotype level, only T. hamatum showed a gradient distribution from south to north in correspondence analysis among the four dominant species. The above results would contribute to the

Expression profiles of putative defence-related proteins in oil palm (Elaeis guineensis) colonized by Ganoderma boninense.

PubMed

Tan, Yung-Chie; Yeoh, Keat-Ai; Wong, Mui-Yun; Ho, Chai-Ling

2013-11-01

Basal stem rot (BSR) is a major disease of oil palm caused by a pathogenic fungus, Ganoderma boninense. However, the interaction between the host plant and its pathogen is not well characterized. To better understand the response of oil palm to G. boninense, transcript profiles of eleven putative defence-related genes from oil palm were measured by quantitative reverse-transcription (qRT)-PCR in the roots of oil palms treated with G. boninense from 3 to 12 weeks post infection (wpi). These transcripts encode putative Bowman-Birk serine protease inhibitors (EgBBI1 and 2), defensin (EgDFS), dehydrin (EgDHN), early methionine-labeled polypeptides (EgEMLP1 and 2), glycine-rich RNA binding protein (EgGRRBP), isoflavone reductase (EgIFR), metallothionein-like protein (EgMT), pathogenesis-related-1 protein (EgPRP), and type 2 ribosome-inactivating protein (EgT2RIP). The transcript abundance of EgBBI2 increased in G. boninense-treated roots at 3 and 6wpi compared to those of controls; while the transcript abundance of EgBBI1, EgDFS, EgEMLP1, EgMT, and EgT2RIP increased in G. boninense-treated roots at 6 or 12wpi. Meanwhile, the gene expression of EgDHN was up-regulated at all three time points in G. boninense-treated roots. The expression profiles of the eleven transcripts were also studied in leaf samples upon inoculation of G. boninense and Trichoderma harzianum to identify potential biomarkers for early detection of BSR. Two candidate genes (EgEMLP1 and EgMT) that have different profiles in G. boninense-treated leaves compared to those infected by T. harzianum may have the potential to be developed as biomarkers for early detection of G. boninense infection. Copyright © 2013 Elsevier GmbH. All rights reserved.

Polymorphisms in Protein Tyrosine Phosphatase Non-receptor Type 2 and 22 (PTPN2/22) Are Linked to Hyper-Proliferative T-Cells and Susceptibility to Mycobacteria in Rheumatoid Arthritis

PubMed Central

Sharp, Robert C.; Beg, Shazia A.; Naser, Saleh A.

2018-01-01

A shared genetic pre-disposition, chronic inflammation, and treatment with similar biologics between Rheumatoid arthritis (RA) and Crohn's disease (CD) have intrigued us to investigate whether the two disorders share trigger association or possible causation. We hypothesized earlier that Single Nucleotide Polymorphisms (SNPs) in the negative regulators Protein Tyrosine Phosphatase Non-receptor type 2 and 22 (PTPN2/22) lead to a dysregulated immune response, susceptibility to environmental triggers, and continued apoptosis as seen in chronic inflammation in RA and CD. To test the hypothesis, peripheral leukocytes samples from 132 consented subjects were genotyped for 9 SNPs in PTPN2/22 using TaqMan™ genotyping. The effect of the SNPs on PTPN2/22 and IFN-γ expression was determined using real time PCR. T-cell proliferation and response to phytohematoagglutonin (PHA) mitogen and mycobacterial antigens were determined by BrdU proliferation assay. Blood samples were also analyzed for the Mycobacterium avium subspecies paratuberculosis (MAP) IS900 gene by nPCR. Out of 9 SNPs examined, heterozygous (TC) or minor (CC) alleles of PTPN2:rs478582 occurred in 79% RA compared to 60% healthy controls (p-values ≤ 0.05; OR = 2.28). Similarly, heterozygous (GA) or minor (AA) alleles of PTPN22:rs2476601 occurred in 29% RA compared to 6% healthy controls (p-values ≤ 0.05; OR = 5.90). PTPN2/22 expression in RA was decreased by 1.2-fold compared to healthy controls. PTPN2:rs478582 upregulated IFN-γ in RA by 1.5-fold. Combined PTPN2:rs478582 and PTPN22:rs2476601 increased T-cell proliferation by 2.7-fold when treated with PHA. Surprisingly, MAP DNA was detected in 34% of RA samples compared to 8% healthy controls, (p-values ≤ 0.05, OR = 5.74). RA samples with PTPN2:rs478582 and/or PTPN22:rs2476601 were more positive for MAP than samples without polymorphisms. Combined occurrence of PTPN2:rs478582 and PTPN22:rs2476601 in association with the presence of MAP has significantly

Plant growth-promoting bacteria as inoculants in agricultural soils

PubMed Central

de Souza, Rocheli; Ambrosini, Adriana; Passaglia, Luciane M.P.

2015-01-01

Abstract Plant-microbe interactions in the rhizosphere are the determinants of plant health, productivity and soil fertility. Plant growth-promoting bacteria (PGPB) are bacteria that can enhance plant growth and protect plants from disease and abiotic stresses through a wide variety of mechanisms; those that establish close associations with plants, such as the endophytes, could be more successful in plant growth promotion. Several important bacterial characteristics, such as biological nitrogen fixation, phosphate solubilization, ACC deaminase activity, and production of siderophores and phytohormones, can be assessed as plant growth promotion (PGP) traits. Bacterial inoculants can contribute to increase agronomic efficiency by reducing production costs and environmental pollution, once the use of chemical fertilizers can be reduced or eliminated if the inoculants are efficient. For bacterial inoculants to obtain success in improving plant growth and productivity, several processes involved can influence the efficiency of inoculation, as for example the exudation by plant roots, the bacterial colonization in the roots, and soil health. This review presents an overview of the importance of soil-plant-microbe interactions to the development of efficient inoculants, once PGPB are extensively studied microorganisms, representing a very diverse group of easily accessible beneficial bacteria. PMID:26537605

Cross inoculation of anthracnose pathogens infecting various tropical fruits

NASA Astrophysics Data System (ADS)

Suparman; Rahmiyah, M.; Pujiastuti, Y.; Gunawan, B.; Arsi

2018-01-01

Anthracnose disease is very important disease of tropical fruits causing significant yield losses. The disease is caused by Colletotrichum spp. and infects almost all tropical fruit species, especially the succulent ones. Various species of Colletotrichum infect various tropical fruits and there are possibilities for cross inoculation to occur among tropical fruits which might cause severe infection. An experimental research was conducted to examine the effect of cross inoculation of anthracnose pathogen among papaya, eggplant, chili and common bean on the infection development and severity of the disease on each inoculated fruit species. Colletotrichum spp. were isolated from naturally infected papaya, eggplant, chili and common bean. Each fungal isolate was purified and identified to determine the species name. The spores of each isolate were then used to separately inoculate healthy and sterilized papaya, eggplant, chili and common bean. The results showed that cross infection developed on chili, eggplant and papaya but not on bean. Chili showed the highest susceptibility to all Colletotrichum isolates and significantly different from eggplant and papaya. The anthracnose pathogen isolated from common bean showed no pathogenicity to other hosts and might be used as cross protection inoculant to the disease in the other hosts.

More on Inoculating Against Reactance to Persuasive Health Messages: The Paradox of Threat.

PubMed

Richards, Adam S; Banas, John A; Magid, Yoav

2017-07-01

This research examined the efficacy of inoculation as a strategy to mitigate psychological reactance based on the level of threat communicated in the forewarning and subsequent persuasive health appeal. Two 2 (inoculation) × 2 (freedom-threatening language) experiments were conducted. The first (N = 181) used elaborated inoculation designed to enhance the threat of impending reactance to a message advocating for responsible alcohol consumption. The second (N = 159) used limited inoculation designed to minimize the threat of impending reactance to a message advocating for responsible soft drink consumption. Results showed that elaborated inoculation increased reactance, whereas limited inoculation decreased reactance but only when the subsequent appeal used less freedom-threatening language. These findings suggest that inoculation has the potential to facilitate or buffer reactance depending on the level of threat communicated in inoculation forewarnings and in subsequent persuasive health appeals.

Increase of fruity aroma during mixed T. delbrueckii/S. cerevisiae wine fermentation is linked to specific esters enhancement.

PubMed

Renault, Philippe; Coulon, Joana; de Revel, Gilles; Barbe, Jean-Christophe; Bely, Marina

2015-08-17

The aim of this work was to study ester formation and the aromatic impact of Torulaspora delbrueckii when used in association with Saccharomyces cerevisiae during the alcoholic fermentation of must. In order to evaluate the influence of the inoculation procedure, sequential and simultaneous mixed cultures were carried out and compared to pure cultures of T. delbrueckii and S. cerevisiae. Our results showed that mixed inoculations allowed the increase, in comparison to S. cerevisiae pure culture, of some esters specifically produced by T. delbrueckii and significantly correlated to the maximal T. delbrueckii population reached in mixed cultures. Thus, ethyl propanoate, ethyl isobutanoate and ethyl dihydrocinnamate were considered as activity markers of T. delbrueckii. On the other hand, isobutyl acetate and isoamyl acetate concentrations were systematically increased during mixed inoculations although not correlated with the development of either species but were rather due to positive interactions between these species. Favoring T. delbrueckii development when performing sequential inoculation enhanced the concentration of esters linked to T. delbrueckii activity. On the contrary, simultaneous inoculation restricted the growth of T. delbrueckii, limiting the production of its activity markers, but involved a very important production of numerous esters due to more important positive interactions between species. These results suggest that the ester concentrations enhancement via interactions during mixed modalities was due to S. cerevisiae production in response to the presence of T. delbrueckii. Finally, sensory analyses showed that mixed inoculations between T. delbrueckii and S. cerevisiae allowed to enhance the complexity and fruity notes of wine in comparison to S. cerevisiae pure culture. Furthermore, the higher levels of ethyl propanoate, ethyl isobutanoate, ethyl dihydrocinnamate and isobutyl acetate in mixed wines were found responsible for the increase of

Experimental inoculation of equine coronavirus into Japanese draft horses.

PubMed

Nemoto, Manabu; Oue, Yasuhiro; Morita, Yoshinori; Kanno, Toru; Kinoshita, Yuta; Niwa, Hidekazu; Ueno, Takanori; Katayama, Yoshinari; Bannai, Hiroshi; Tsujimura, Koji; Yamanaka, Takashi; Kondo, Takashi

2014-12-01

Recently, outbreaks associated with equine coronavirus (ECoV) have occurred in Japan and the United States. While ECoV is likely to be pathogenic to horses, it has not been shown that experimental inoculation of horses with ECoV produces clinical signs of disease. In this study, we inoculated three Japanese draft horses with an ECoV-positive diarrheic fecal sample to confirm infection after inoculation and to investigate the clinical course and virus shedding patterns of ECoV. Virus neutralization tests showed that all three horses became infected with ECoV. Two of the three horses developed clinical signs similar to those observed during ECoV outbreaks, including fever, anorexia, and gastrointestinal dysfunction. All horses excreted a large amount of virus into their feces for more than 9 days after inoculation regardless of the presence or absence of clinical signs, which suggests that feces are an important source of ECoV infection. ECoV was also detected in nasal swabs from all horses, suggesting that respiratory transmission of ECoV may occur. Both symptomatic horses developed viremia, while the asymptomatic horse did not. White blood cell counts and serum amyloid A concentrations changed relative to the clinical condition of the inoculated horses; these may be useful markers for monitoring the clinical status of horses infected with ECoV. This is the first report of induction of clinical signs of ECoV infection in horses by experimental inoculation. These clinical and virological findings should aid further investigation of the pathogenesis of ECoV.

Inoculation of soil native cyanobacteria to restore arid degraded soils

NASA Astrophysics Data System (ADS)

Raúl Román Fernández, José; Roncero Ramos, Beatriz; Chamizo de la Piedra, Sonia; Rodríguez Caballero, Emilio; Ángeles Muñoz Martín, M.; Mateo, Pilar; Cantón Castilla, Yolanda

2017-04-01

Restoration projects in semiarid lands often yield poor results. Water scarcity, low soil fertility, and poor soil structure strongly limit the survival and growth of planted seedlings in these areas. Under these conditions, a previous stage that improves edaphic conditions would turn out to a successful plant restoration. By successfully colonizing arid soils, cyanobacteria naturally provide suitable edaphic conditions, enhancing water availability, soil fertility and soil stability. Furthermore, cyanobacteria can be easily isolated and cultured ex-situ to produce high quantities of biomass, representing a potential tool to restore large areas efficiently. The objective of this study was to test the effect of inoculated cyanobacteria on degraded soils at three different semiarid areas from southeast Spain: Tabernas badlands, a limestone quarry located in Gádor, and grazed grassland in Las Amoladeras (Cabo de Gata). Soil native cyanobacteria belonging to three representative N-fixing genera (Nostoc, Scytonema and Tolypothrix) were isolated from such soils and cultured in BG110 medium. Each strain was inoculated (6 g m-2), separately and mixed (all in the same proportion), on Petri dishes with 80 g of each soil. Biocrust development was monitored during 3 months in these soils under laboratory conditions, at a constant temperature of 25oC. During the experiment, two irrigation treatments were applied simulating a dry (180 mm) and a wet (360 mm) rainfall year (average recorded in the study sites). After 3 months, net CO2 flux, spectral response and soil surface microtopography (1 mm spatial resolution) of inoculated and control soils was measured under wet conditions, all of them as a surrogate of biocrust development. Samples of the surface crust were collected in order to determine total soil organic carbon (SOC) content. The inoculated soils showed positive values of net CO2 flux, thus indicating a net CO2 uptake, whereas control soils showed CO2 fluxes closed to

The phosphatase JKAP/DUSP22 inhibits T-cell receptor signalling and autoimmunity by inactivating Lck.

PubMed

Li, Ju-Pi; Yang, Chia-Yu; Chuang, Huai-Chia; Lan, Joung-Liang; Chen, Der-Yuan; Chen, Yi-Ming; Wang, Xiaohong; Chen, Alice J; Belmont, John W; Tan, Tse-Hua

2014-04-09

JNK pathway-associated phosphatase (JKAP, also known as DUSP22 or JSP-1) is a JNK activator. The in vivo role of JKAP in immune regulation remains unclear. Here we report that JKAP directly inactivates Lck by dephosphorylating tyrosine-394 residue during T-cell receptor (TCR) signalling. JKAP-knockout T cells display enhanced cell proliferation and cytokine production. JKAP-knockout mice show enhanced T-cell-mediated immune responses and are more susceptible to experimental autoimmune encephalomyelitis (EAE). In addition, the recipient mice that are adoptively transferred with JKAP-knockout T cells show exacerbated EAE symptoms. Aged JKAP-knockout mice spontaneously develop inflammation and autoimmunity. Thus, our results indicate that JKAP is an important phosphatase that inactivates Lck in the TCR signalling turn-off stage, leading to suppression of T-cell-mediated immunity and autoimmunity.

Integrated control of wood destroying basidiomycetes combining Cu-based wood preservatives and Trichoderma spp.

PubMed Central

2017-01-01

The production of new generation of wood preservatives (without addition of a co-biocide) in combination with an exchange of wood poles on identical sites with high fungal inoculum, has resulted in an increase of premature failures of wood utility poles in the last decades. Wood destroying basidiomycetes inhabiting sites where poles have been installed, have developed resistance against wood preservatives. The objective of the in vitro studies was to identify a Trichoderma spp. with a highly antagonistic potential against wood destroying basidiomycetes that is capable of colonizing Cu-rich environments. For this purpose, the activity of five Trichoderma spp. on Cu-rich medium was evaluated according to its growth and sporulation rates. The influence of the selected Trichoderma spp. on wood colonization and degradation by five wood destroying basidiomycetes was quantitatively analyzed by means of dry weight loss of wood specimens. Furthermore, the preventative effect of the selected Trichoderma spp. in combination with four Cu-based preservatives was also examined by mass loss and histological changes in the wood specimens. Trichoderma harzianum (T-720) was considered the biocontrol agent with higher antagonistic potential to colonize Cu-rich environments (up to 0.1% CuSO4 amended medium). T. harzianum demonstrated significant preventative effect on wood specimens against four wood destroying basidiomycetes. The combined effect of T. harzianum and Cu-based wood preservatives demonstrated that after 9 months incubation with two wood destroying basidiomycetes, wood specimens treated with 3.8 kg m-3 copper-chromium had weight losses between 55–65%, whereas containers previously treated with T. harzianum had significantly lower weight losses (0–25%). Histological studies on one of the wood destroying basidiomycetes revealed typical decomposition of wood cells by brown-rot fungi in Cu-impregnated samples, that were notably absent in wood specimens previously exposed

Integrated control of wood destroying basidiomycetes combining Cu-based wood preservatives and Trichoderma spp.

PubMed

Ribera, Javier; Fink, Siegfried; Bas, Maria Del Carmen; Schwarze, Francis W M R

2017-01-01

The production of new generation of wood preservatives (without addition of a co-biocide) in combination with an exchange of wood poles on identical sites with high fungal inoculum, has resulted in an increase of premature failures of wood utility poles in the last decades. Wood destroying basidiomycetes inhabiting sites where poles have been installed, have developed resistance against wood preservatives. The objective of the in vitro studies was to identify a Trichoderma spp. with a highly antagonistic potential against wood destroying basidiomycetes that is capable of colonizing Cu-rich environments. For this purpose, the activity of five Trichoderma spp. on Cu-rich medium was evaluated according to its growth and sporulation rates. The influence of the selected Trichoderma spp. on wood colonization and degradation by five wood destroying basidiomycetes was quantitatively analyzed by means of dry weight loss of wood specimens. Furthermore, the preventative effect of the selected Trichoderma spp. in combination with four Cu-based preservatives was also examined by mass loss and histological changes in the wood specimens. Trichoderma harzianum (T-720) was considered the biocontrol agent with higher antagonistic potential to colonize Cu-rich environments (up to 0.1% CuSO4 amended medium). T. harzianum demonstrated significant preventative effect on wood specimens against four wood destroying basidiomycetes. The combined effect of T. harzianum and Cu-based wood preservatives demonstrated that after 9 months incubation with two wood destroying basidiomycetes, wood specimens treated with 3.8 kg m-3 copper-chromium had weight losses between 55-65%, whereas containers previously treated with T. harzianum had significantly lower weight losses (0-25%). Histological studies on one of the wood destroying basidiomycetes revealed typical decomposition of wood cells by brown-rot fungi in Cu-impregnated samples, that were notably absent in wood specimens previously exposed to T

Metabolites of Trichoderma species isolated from damp building materials.

PubMed

McMullin, David R; Renaud, Justin B; Barasubiye, Tharcisse; Sumarah, Mark W; Miller, J David

2017-07-01

Buildings that have been flooded often have high concentrations of Trichoderma spores in the air while drying. Inhaled spores and spore and mycelial fragments contain large amounts of fungal glucan and natural products that contribute to the symptoms associated with indoor mould exposures. In this study, we considered both small molecules and peptaibol profiles of T. atroviride, T. koningiopsis, T. citrinoviride, and T. harzianum strains obtained from damp buildings in eastern Canada. Twenty-residue peptaibols and sorbicillin-derived metabolites (1-6) including a new structure, (R)-vertinolide (1), were characterized from T. citrinoviride. Trichoderma koningiopsis produced several koninginins (7-10), trikoningin KA V, and the 11-residue lipopeptaibols trikoningin KB I and trikoningin KB II. Trichoderma atroviride biosynthesized a mixture of 19-residue trichorzianine-like peptaibols, whereas T. harzianum produced 18-residue trichokindin-like peptaibols and the 11-residue harzianin HB I that was subsequently identified from the studied T. citrinoviride strain. Two α-pyrones, 6-pentyl-pyran-2-one (11) and an oxidized analog (12), were produced by both T. atroviride and T. harzianum. Aside from exposure to low molecular weight natural products, inhalation of Trichoderma spores and mycelial fragments may result in exposure to membrane-disrupting peptaibols. This investigation contributes to a more comprehensive understanding of the biologically active natural products produced by fungi commonly found in damp buildings.

Survival of Campylobacter jejuni inoculated into ground beef.

PubMed Central

Stern, N J; Kotula, A W

1982-01-01

Ground beef was inoculated with mixed cultures of Campylobacter jejuni, and the samples were subjected to various cooking and cold-storage temperatures. When samples were heated in an oven at either 190 or 218 degrees C, approximately 10(7) cells of C. jejuni per g were inactivated (less than 30 cells per g) in less than 10 min after the ground beef reached an internal temperature of 70 degrees C. When the samples were held at -15 degrees C over 14 days of storage, the numbers of C. jejuni declined by 3 log10. When inoculated samples were stored with an equal amount of Cary-Blair diluent at 4 degrees C, no changes in viability were observed over 14 days of storage. Twenty-five times as much C. jejuni was recovered from inoculated ground beef when either 10% glycerol or 10% dimethyl sulfoxide was added to an equal amount of ground beef before freezing as was recovered from peptone-diluted ground beef. Twice as much inoculated C. jejuni was recovered from ground beef plus Cary-Blair diluent as was recovered from ground beef plus peptone diluent. PMID:7181502

Outcome of oral infection in mice inoculated with Trypanosoma cruzi IV of the Western Brazilian Amazon.

PubMed

Margioto Teston, Ana Paula; de Abreu, Ana Paula; Abegg, Camila Piva; Gomes, Mônica Lúcia; de Ornelas Toledo, Max Jean

2017-02-01

A new epidemiological view of American trypanosomiasis or Chagas disease has been formulated in recent decades. Oral transmission of the etiological agent of Chagas disease, Trypanosoma cruzi, has been the most common form of transmission. The T. cruzi discrete typing units TcI and TcIV have been involved in tens outbreaks of acute cases of Chagas disease in the Brazilian Amazon region. We investigated the intensity of infection in mice that were orally inoculated (OR group) with four strains of TcIV that were isolated from two outbreaks of acute Chagas disease that was orally acquired in the state of Amazonas, Brazil. We compared the OR group with mice that were intraperitoneally inoculated (IP group). Blood samples were analyzed by fresh blood examination, hemoculture, and conventional and qualitative real-time polymerase chain reaction (PCR). Samples of different tissues were analyzed by quantitative real-time PCR. The OR group exhibited a higher maximum peak of parasitemia, greater rates of positivity, and higher parasite loads in different tissues during acute infection compared with the IP group, indicating a greater intensity of orally acquired infection. Mice that were orally inoculated with TcIV strains that were obtained from two outbreaks of orally acquired Chagas disease in Amazonas, Brazil, exhibited a more intense course of infection compared with intraperitoneally inoculated mice, reflected by higher levels of parasitemia and parasite loads. Copyright © 2016. Published by Elsevier B.V.

HFB7 - A novel orphan hydrophobin of the Harzianum and Virens clades of Trichoderma, is involved in response to biotic and abiotic stresses.

PubMed

Przylucka, Agnes; Akcapinar, Gunseli Bayram; Chenthamara, Komal; Cai, Feng; Grujic, Marica; Karpenko, Juriy; Livoi, Miriam; Shen, Qirong; Kubicek, Christian P; Druzhinina, Irina S

2017-05-01

Hydrophobins are small secreted cysteine-rich proteins exclusively found in fungi. They are able to self-assemble in single molecular layers at hydrophobic-hydrophilic interfaces and can therefore be directly involved in establishment of fungi in their habitat. The genomes of filamentous mycotrophic fungi Trichoderma encode a rich diversity of hydrophobins, which are divided in several groups based on their structure and evolution. Here we describe a new member of class II hydrophobins, HFB7, that has a taxonomically restricted occurrence in Harzianum and Virens clades of Trichoderma. Evolutionary analysis reveals that HFB7 proteins form a separate clade distinct from other Trichoderma class II hydrophobins and that genes encoding them evolve under positive selection pressure. Homology modelling of HFB7 structure in comparison to T. reesei HFB2 reveals that the two large hydrophobic patches on the surface of the protein are remarkably conserved between the two hydrophobins despite significant difference in their primary structures. Expression of hfb7 gene in T. virens increases at interactions with other fungi and a plant and in response to a diversity of abiotic stress conditions, and is also upregulated during formation of aerial mycelium in a standing liquid culture. This upregulation significantly exceeds that of expression of hfb7 under a strong constitutive promoter, and T. virens strains overexpressing hfb7 thus display only changes in traits characterized by low hfb7 expression, i.e. faster growth in submerged liquid culture. The hfb7 gene is not expressed in conidia. Our data allow to conclude that this protein is involved in defence of Trichoderma against a diversity of stress factors related to the oxidative stress. Moreover, HFB7 likely helps in the establishment of the fungus in wetlands or other conditions related to high humidity. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Temperature control strategy to enhance the activity of yeast inoculated into compost raw material for accelerated composting.

PubMed

Nakasaki, Kiyohiko; Hirai, Hidehira

2017-07-01

The effects of inoculating the mesophilic yeast Pichia kudriavzevii RB1, which is able to degrade organic acids, on organic matter degradation in composting were elucidated. When model food waste with high carbohydrate content (C/N=22.3) was used, fluctuation in the inoculated yeast cell density was observed, as well as fluctuation in the composting temperature until day 5 when the temperature rose to 60°C, which is lethal for the yeast. After the decrease in yeast, acetic acid accumulated to levels as high as 20mg/g-ds in the composting material and vigorous organic matter degradation was inhibited. However, by maintaining the temperature at 40°C for 2days during the heating phase in the early stage of composting, both the organic acids originally contained in the raw material and acetic acid produced during the heating phase were degraded by the yeast. The concentration of acetic acid was kept at a relatively low level (10.1mg/g-ds at the highest), thereby promoting the degradation of organic matter by other microorganisms and accelerating the composting process. These results indicate that temperature control enhances the effects of microbial inoculation into composts. Copyright © 2017 Elsevier Ltd. All rights reserved.

Drop of egg production in chickens by experimental infection with an avian metapneumovirus strain PLE8T1 derived from swollen head syndrome and the application to evaluate vaccine.

PubMed

Sugiyama, Miki; Koimaru, Hiroyuki; Shiba, Masahiro; Ono, Eriko; Nagata, Tadashi; Ito, Toshihiro

2006-08-01

Decreases in egg production and increased incidence of abnormal eggs due to malformation of egg shells were observed in specific pathogen free (SPF) 173-day-old laying hens inoculated intravenously with an avian metapneumovirus (aMPV) strain PLE8T1. This strain was derived from an isolate from broiler birds exhibiting swollen head syndrome (SHS). Some SPF birds inoculated with the virus showed, slight diarrhea without any respiratory symptoms. Thus, the PLE8T1 strain was used as a challenge virus to evaluate efficacy of aMPV vaccines. SPF chickens which received a live attenuated aMPV vaccine (NEMOVAC; Merial) at 7 or 77 days old and an inactivated aMPV vaccine (OVO-4; Merial) at 105 days old were protected against poor egg production caused by the challenge with the PLE8T1 strain. Thus, aMPV, the PLE8T1 strain passaged 22 times after isolation, from birds exhibiting SHS, could induce a drop in egg production in laying hens accompanied by malformation of egg shells. It was suggested that this challenge system could be applied to evaluate the efficacy of aMPV vaccine.

Differential growth response of various crop species to arbuscular mycorrhizal inoculation.

PubMed

Eo, Ju-Kyeong; Eom, Ahn-Heum

2009-03-01

To investigate the growth response of various crop species to mycorrhizal inoculation, arbuscular mycorrhizal fungi were applied to Glycine max, Vigna angularis, Senna tora, Hordeum vulgare var. hexastichon. Zea mays, Sorghum bicolor, Allium tuberosum, Solanum melongena, and Capsicum annuum. The biomass of the inoculated crops was measured every two weeks for the 12-week growth period. By measuring biomass, we calculated the mycorrhizal responsiveness of the nine crop species. Among the nine crop species, four species showed a significant response to mycorrhizal inoculation. The shoot biomasses of V. angularis, C. annuum, A. tuberosum, and S. tora significantly increased with mycorrhizal inoculation.

Large numbers of interleukins-22- and -17A-producing T helper cells in cholangiocarcinoma related to liver fluke infection.

PubMed

Su, Si-Biao; Zhang, Jian-Feng; Huang, Fei-Fei; Cen, Yu; Jiang, Hai-Xing

2017-08-01

Cholangiocarcinoma (CCA) associated with liver fluke infection involves inflammatory and immune processes; however, whether these involve the proinflammatory cytokine IL-17A and proliferative cytokine IL-22 remains unclear. Here, numbers of IL-22- and IL-17A-producing Th cells and cytokine concentrations in 30 patients with CCA and long-term liver fluke infection, 40 patients with liver-fluke infection but not CCA, and 16 healthy controls were compared. Analyses were performed using immunohistochemistry, flow cytometry, ELISA and RT-PCR. Immunohistochemical staining showed weaker expression of IL-22 and IL-17A in patients with CCA with than in those without liver fluke infection (P < 0.01). Flow cytometry revealed significantly greater median proportions of IL-22-producing T helper cells in patients with CCA (2.2%) than in those without it (0.69%) or controls (0.4%, P < 0.001). Similar results were obtained for IL-17A-producing T helper cells. ELISA revealed plasma concentrations of IL-22 were 1.3-fold higher in patients with CCA than in those without it and 4.6-fold higher than in controls (P < 0.001). Plasma concentrations of IL-17A were 2.5-fold higher in patients with CCA than in those without it, and 21-fold higher than in controls (P < 0.001). Amounts of IL-22 and IL-17A mRNAs in blood were significantly higher in patients with CCA than in the other two groups. Proportions of CD4 + CD45RO + T cells producing IL-22 correlated with proportions producing IL-17A (r = 0.759; P < 0.001), and plasma concentrations of IL-22 correlated with those of IL-17A (r = 0.726; P < 0.001). These results suggest that both IL-17A and IL-22 affect development of CCA related to liver fluke infection. © 2017 The Societies and John Wiley & Sons Australia, Ltd.

Screening and identification of resistance related proteins from apple leaves inoculated with Marssonina coronaria (EII. & J. J. Davis)

PubMed Central

2014-01-01

Background Apple, an invaluable fruit crop worldwide, is often prone to infection by pathogenic fungi. Identification of potentially resistance-conferring apple proteins is one of the most important aims for studying apple resistance mechanisms and promoting the development of disease-resistant apple strains. In order to find proteins which promote resistance to Marssonina coronaria, a deadly pathogen which has been related to premature apple maturation, proteomes from apple leaves inoculated with M. coronaria were characterized at 3 and 6 days post-inoculation by two dimensional electrophoresis (2-DE). Results Overall, 59 differentially accumulated protein spots between inoculation and non-inoculation were successfully identified and aligned as 35 different proteins or protein families which involved in photosynthesis, amino acid metabolism, transport, energy metabolism, carbohydrate metabolism, binding, antioxidant, defense and stress. Quantitative real-time PCR (qRT-PCR) was also used to examine the change of some defense and stress related genes abundance under inoculated conditions. Conclusions In a conclusion, different proteins in response to Marssonina coronaria were identified by proteomic analysis. Among of these proteins, there are some PR proteins, for example class III endo-chitinase, beta-1,3-glucanase and thaumatine-like protein, and some antioxidant related proteins including aldo/keto reductase AKR, ascorbate peroxidase and phi class glutathione S-transferase protein that were associated with disease resistance. The transcription levels of class III endo-chitinase (L13) and beta-1, 3-glucanase (L17) have a good relation with the abundance of the encoded protein’s accumulation, however, the mRNA abundance of thaumatine-like protein (L22) and ascorbate peroxidase (L28) are not correlated with their protein abundance of encoded protein. To elucidate the resistant mechanism, the data in the present study will promote us to investigate further the

Inheritance of balanced translocation t(17; 22) from a Down syndrome mother to a phenotypically normal daughter.

PubMed

Liu, X Y; Jiang, Y T; Wang, R X; Luo, L L; Liu, Y H; Liu, R Z

2015-08-28

We report that a 30-year-old woman with mental retardation was referred for prenatal diagnoses during pregnancy. An ultrasound scan showed that the heart structure and function of the fetus were normal. Cytogenetic analysis showed that the female karyotype was 47,XX, t(17; 22) (q21; q11), +21. The woman's husband had a normal male karyotype and was phenotypically normal. During this first pregnancy, an amniocentesis, which was done at 19 weeks, revealed that the fetal karyotype was 46,XX, t(17; 22) (q21; q11). Fluorescence in situ hybridization testing of amniotic fluid gave a normal result for chromosome 21. The child was a phenotypically normal female baby.

Using common mycorrhizal networks for controlled inoculation of Quercus spp. with Tuber melanosporum: the nurse plant method.

PubMed

Pereira, Guillermo; Palfner, Götz; Chávez, Daniel; Suz, Laura M; Machuca, Angela; Honrubia, Mario

2013-07-01

The high cost and restricted availability of black truffle spore inoculum for controlled mycorrhiza formation of host trees produced for truffle orchards worldwide encourage the search for more efficient and sustainable inoculation methods that can be applied globally. In this study, we evaluated the potential of the nurse plant method for the controlled inoculation of Quercus cerris and Quercus robur with Tuber melanosporum by mycorrhizal networks in pot cultures. Pine bark compost, adjusted to pH 7.8 by liming, was used as substrate for all assays. Initially, Q. robur seedlings were inoculated with truffle spores and cultured for 12 months. After this period, the plants presenting 74 % mycorrhizal fine roots were transferred to larger containers. Nurse plants were used for two treatments of two different nursling species: five sterilized acorns or five 45-day-old, axenically grown Q. robur or Q. cerris seedlings, planted in containers around the nurse plant. After 6 months, colonized nursling plant root tips showed that mycorrhiza formation by T. melanosporum was higher than 45 % in the seedlings tested, with the most successful nursling combination being Q. cerris seedlings, reaching 81 % colonization. Bulk identification of T. melanosporum mycorrhizae was based on morphological and anatomical features and confirmed by sequencing of the internal transcribed spacer region of the ribosomal DNA of selected root tips. Our results show that the nurse plant method yields attractive rates of mycorrhiza formation by the Périgord black truffle and suggest that establishing and maintaining common mycorrhizal networks in pot cultures enables sustained use of the initial spore inoculum.

Inoculation Stress Hypothesis of Environmental Enrichment

PubMed Central

Crofton, Elizabeth J.; Zhang, Yafang; Green, Thomas A.

2014-01-01

One hallmark of psychiatric conditions is the vast continuum of individual differences in susceptibility vs. resilience resulting from the interaction of genetic and environmental factors. The environmental enrichment paradigm is an animal model that is useful for studying a range of psychiatric conditions, including protective phenotypes in addiction and depression models. The major question is how environmental enrichment, a non-drug and non-surgical manipulation, can produce such robust individual differences in such a wide range of behaviors. This paper draws from a variety of published sources to outline a coherent hypothesis of inoculation stress as a factor producing the protective enrichment phenotypes. The basic tenet suggests that chronic mild stress from living in a complex environment and interacting non-aggressively with conspecifics can inoculate enriched rats against subsequent stressors and/or drugs of abuse. This paper reviews the enrichment phenotypes, mulls the fundamental nature of environmental enrichment vs. isolation, discusses the most appropriate control for environmental enrichment, and challenges the idea that cortisol/corticosterone equals stress. The intent of the inoculation stress hypothesis of environmental enrichment is to provide a scaffold with which to build testable hypotheses for the elucidation of the molecular mechanisms underlying these protective phenotypes and thus provide new therapeutic targets to treat psychiatric/neurological conditions. PMID:25449533

The Application of Stress-Relaxation Test to Life Assessment of T911/T22 Weld Metal

NASA Astrophysics Data System (ADS)

Cao, Tieshan; Zhao, Jie; Cheng, Congqian; Li, Huifang

2016-03-01

A dissimilar weld metal was obtained through submerged arc welding of a T911 steel to a T22 steel, and its creep property was explored by stress-relaxation test assisted by some conventional creep tests. The creep rate information of the stress-relaxation test was compared to the minimum and the average creep rates of the conventional creep test. Log-log graph showed that the creep rate of the stress-relaxation test was in a linear relationship with the minimum creep rate of the conventional creep test. Thus, the creep rate of stress-relaxation test could be used in the Monkman-Grant relation to calculate the rupture life. The creep rate of the stress-relaxation test was similar to the average creep rate, and thereby the rupture life could be evaluated by a method of "time to rupture strain." The results also showed that rupture life which was assessed by the Monkman-Grant relation was more accurate than that obtained through the method of "time to rupture strain."

Transmission of a t(13q22q) chromosome observed in three generations with segregation of the translocation D1-trisomy syndrome.

PubMed

Abe, T; Morita, M; Kawai, K; Misawa, S; Kanai, H; Hirose, G; Fujita, H

1975-09-20

A case of an inherited type of D/G translocation D1-trisomy syndrome was described. A female proposita who had the clinical signs of D1-trisomy syndrome was found to have a chromosome complement of 46,XX,--G,+t(DqGq). examination of Q- and G-stained karyotypes revealed that the chromosomes involved in the translocation were members of Nos. 13 and 22, or t(13q22q) with breaks at p12 of both chromosomes. C-stained figures also showed a large heterochromatin block in its centromeric region. The t(13q22q) chromosome was transmitted from the paternal grandmother of the proposita through at least three generations.

Meta-analysis reveals PTPN22 1858C/T polymorphism confers susceptibility to rheumatoid arthritis in Caucasian but not in Asian population.

PubMed

Nabi, Gowher; Akhter, Naseem; Wahid, Mohd; Bhatia, Kanchan; Mandal, Raju Kumar; Dar, Sajad Ahmad; Jawed, Arshad; Haque, Shafiul

2016-01-01

The PTPN22 1858C/T polymorphism is associated with rheumatoid arthritis (RA). However, reports from the Asian populations are conflicting in nature and lacks consensus. The aim of our study was to evaluate the association between the PTPN22 1858C/T polymorphism and RA in Asian and Caucasian subjects by carrying out a meta-analysis of Asian and Caucasian data. A total of 27 205 RA cases and 27 677 controls were considered in the present meta-analysis involving eight Asian and 35 Caucasian studies. The pooled odds ratios (ORs) were performed for the allele, dominant, and recessive genetic model. No statistically significant association was found between the PTPN22 1858C/T polymorphism and risk of RA in Asian population (allele genetic model: OR = 1.217, 95% confidence interval (CI) = 0.99-1.496, p value 0.061; dominant genetic model: OR = 1.238, 95% CI = 0.982-1.562, p value 0.071; recessive genetic model: OR = 1.964, 95% CI = 0.678-5.693, p value 0.213). A significant association with risk of RA in Caucasian population suggesting that T-- allele does confer susceptibility to RA in this subgroup was observed (allele genetic model: OR = 1.638, 95% CI = 1.574-1.705, p value < 0.0001; dominant genetic model: OR = 1.67, 95% CI = 1.598-1.745, p value < 0.0001; recessive genetic model: OR = 2.65, 95% CI = 2.273-3.089, p value < 0.0001). The PTPN22 1858C/T polymorphism is not associated with RA risk in Asian populations. However, our meta-analysis confirms that the PTPN22 1858C/T polymorphism is associated with RA susceptibility in Caucasians.

The constitutional t(11;22): implications for a novel mechanism responsible for gross chromosomal rearrangements

PubMed Central

Kurahashi, H; Inagaki, H; Ohye, T; Kogo, H; Tsutsumi, M; Kato, T; Tong, M; Emanuel, BS

2012-01-01

The constitutional t(11;22)(q23;q11) is the most common recurrent non-Robertsonian translocation in humans. The breakpoint sequences of both chromosomes are characterized by several hundred base pairs of palindromic AT-rich repeats (PATRRs). Similar PATRRs have also been identified at the breakpoints of other nonrecurrent translocations, suggesting that PATRR-mediated chromosomal translocation represents one of the universal pathways for gross chromosomal rearrangement in the human genome. We propose that PATRRs have the potential to form cruciform structures through intrastrand-base pairing in single-stranded DNA, creating a source of genomic instability and leading to translocations. Indeed, de novo examples of the t(11;22) are detected at a high frequency in sperm from normal healthy males. This review synthesizes recent data illustrating a novel paradigm for an apparent spermatogenesis-specific translocation mechanism. This observation has important implications pertaining to the predominantly paternal origin of de novo gross chromosomal rearrangements in humans. PMID:20507342

CD22-targeted CAR T cells induce remission in B-ALL that is naive or resistant to CD19-targeted CAR immunotherapy.

PubMed

Fry, Terry J; Shah, Nirali N; Orentas, Rimas J; Stetler-Stevenson, Maryalice; Yuan, Constance M; Ramakrishna, Sneha; Wolters, Pamela; Martin, Staci; Delbrook, Cindy; Yates, Bonnie; Shalabi, Haneen; Fountaine, Thomas J; Shern, Jack F; Majzner, Robbie G; Stroncek, David F; Sabatino, Marianna; Feng, Yang; Dimitrov, Dimiter S; Zhang, Ling; Nguyen, Sang; Qin, Haiying; Dropulic, Boro; Lee, Daniel W; Mackall, Crystal L

2018-01-01

Chimeric antigen receptor (CAR) T cells targeting CD19 mediate potent effects in relapsed and/or refractory pre-B cell acute lymphoblastic leukemia (B-ALL), but antigen loss is a frequent cause of resistance to CD19-targeted immunotherapy. CD22 is also expressed in most cases of B-ALL and is usually retained following CD19 loss. We report results from a phase 1 trial testing a new CD22-targeted CAR (CD22-CAR) in 21 children and adults, including 17 who were previously treated with CD19-directed immunotherapy. Dose-dependent antileukemic activity was observed, with complete remission obtained in 73% (11/15) of patients receiving ≥1 × 10 6 CD22-CAR T cells per kg body weight, including 5 of 5 patients with CD19 dim or CD19 - B-ALL. Median remission duration was 6 months. Relapses were associated with diminished CD22 site density that likely permitted CD22 + cell escape from killing by CD22-CAR T cells. These results are the first to establish the clinical activity of a CD22-CAR in B-ALL, including leukemia resistant to anti-CD19 immunotherapy, demonstrating potency against B-ALL comparable to that of CD19-CAR at biologically active doses. Our results also highlight the critical role played by antigen density in regulating CAR function.

Growth and Yield Responses of Cowpea to Inoculation and Phosphorus Fertilization in Different Environments

PubMed Central

Kyei-Boahen, Stephen; Savala, Canon E. N.; Chikoye, David; Abaidoo, Robert

2017-01-01

Cowpea (Vigna unguiculata) is a major source of dietary protein and essential component of the cropping systems in semi-arid regions of Sub-Saharan Africa. However, yields are very low due to lack of improved cultivars, poor management practices, and limited inputs use. The objectives of this study were to assess the effects of rhizobia inoculant and P on nodulation, N accumulation and yield of two cowpea cultivars in Mozambique. Field study was conducted in three contrasting environments during the 2013/2014 and 2014/2015 seasons using randomized complete block design with four replications and four treatments. Treatments consisted of seed inoculation, application of 40 kg P2O5 ha-1, inoculation + P, and a non-inoculated control. The most probable number (MPN) technique was used to estimate the indigenous bradyrhizobia populations at the experimental sites. The rhizobia numbers at the sites varied from 5.27 × 102 to 1.07 × 103 cells g-1 soil. Inoculation increased nodule number by 34–76% and doubled nodule dry weight (78 to 160 mg plant-1). P application improved nodulation and interacted positively with the inoculant. Inoculation, P, and inoculant + P increased shoot dry weight, and shoot and grain N content across locations but increases in number of pods plant-1, seeds pod-1, and 100-seed weight were not consistent among treatments across locations. Shoot N content was consistently high for the inoculated plants and also for the inoculated + P fertilized plants, whereas the non-inoculated control plants had the lowest tissue N content. P uptake in shoot ranged from 1.72 to 3.77 g kg-1 and was higher for plants that received P fertilizer alone. Inoculation and P either alone or in combination consistently increased cowpea grain yield across locations with yields ranging from 1097 kg ha-1 for the non-inoculated control to 1674 kg ha-1 for the inoculant + P treatment. Grain protein concentration followed a similar trend as grain yield and ranged from 223 to

Hypothalamic-pituitary-adrenal axis physiology and cognitive control of behavior in stress inoculated monkeys.

PubMed

Parker, Karen J; Buckmaster, Christine L; Lindley, Steven E; Schatzberg, Alan F; Lyons, David M

2012-01-01

Monkeys exposed to stress inoculation protocols early in life subsequently exhibit diminished neurobiological responses to moderate psychological stressors and enhanced cognitive control of behavior during juvenile development compared to non-inoculated monkeys. The present experiments extended these findings and revealed that stress inoculated monkeys: (a) mount neurobiological responses equivalent to non-inoculated monkeys when the stressor is of sufficient intensity, and (b) continue to exhibit enhanced cognitive control as young adults compared to non-inoculated monkeys. These results suggest that stress inoculation protocols alter the appraisal of and response to moderate stressors as less threatening and permanently enhance cognitive control, at least through early adulthood. These data therefore support the notion that the stress inoculation phenotype reflects stress resilience rather than stress pathology.

Effect of actinobacteria agent inoculation methods on cellulose degradation during composting based on redundancy analysis.

PubMed

Zhao, Yue; Lu, Qian; Wei, Yuquan; Cui, Hongyang; Zhang, Xu; Wang, Xueqin; Shan, Si; Wei, Zimin

2016-11-01

In this study, actinobacteria agent including Streptomyces sp. and Micromonospora sp. were inoculated during chicken manure composting by different inoculation methods. The effect of different treatments on cellulose degradation and the relationship between inoculants and indigenous actinobacteria were investigated during composting. The results showed that inoculation in different stages of composting all improved the actinobacteria community diversity particularly in the cooling stage of composting (M3). Moreover, inoculation could distinctly accelerate the degradation of organic matters (OM) especially celluloses. Redundancy analysis indicated that the correlation between indigenous actinobacteria and degradation of OM and cellulose were regulated by inoculants and there were significant differences between different inoculation methods. Furthermore, synergy between indigenous actinobacteria and inoculants for degradation of OM and cellulose in M3 was better than other treatments. Conclusively, we suggested an inoculation method to regulate the indigenous actinobacteria based on the relationship between inoculants and indigenous actinobacteria and degradation content. Copyright © 2016 Elsevier Ltd. All rights reserved.

The phosphatase JKAP/DUSP22 inhibits t-cell receptor signalling and autoimmunity by inactivating Lck

USDA-ARS?s Scientific Manuscript database

JNK pathway-associated phosphatase (JKAP, also known as DUSP22 or JSP-1) is a JNK activator. The in vivo role of JKAP in immune regulation remains unclear. Here we report that JKAP directly inactivates Lck by dephosphorylating tyrosine-394 residue during T-cell receptor (TCR) signalling. JKAP-knocko...

Establishment and effectiveness of inoculated arbuscular mycorrhizal fungi in agricultural soils.

PubMed

Köhl, Luise; Lukasiewicz, Catherine E; van der Heijden, Marcel G A

2016-01-01

Arbuscular mycorrhizal fungi (AMF) are promoted as biofertilizers for sustainable agriculture. So far, most researchers have investigated the effects of AMF on plant growth under highly controlled conditions with sterilized soil, soil substrates or soils with low available P or low inoculum potential. However, it is still poorly documented whether inoculated AMF can successfully establish in field soils with native AMF communities and enhance plant growth. We inoculated grassland microcosms planted with a grass-clover mixture (Lolium multiflorum and Trifolium pratense) with the arbuscular mycorrhizal fungus Rhizoglomus irregulare. The microcosms were filled with eight different unsterilized field soils that varied greatly in soil type and chemical characteristics and indigenous AMF communities. We tested whether inoculation with AMF enhanced plant biomass and R. irregulare abundance using a species specific qPCR. Inoculation increased the abundance of R. irregulare in all soils, irrespective of soil P availability, the initial abundance of R. irregulare or the abundance of native AM fungal communities. AMF inoculation had no effect on the grass but significantly enhanced clover yield in five out of eight field soils. The results demonstrate that AMF inoculation can be successful, even when soil P availability is high and native AMF communities are abundant. © 2015 John Wiley & Sons Ltd.

Experimental Inoculation of Egyptian Fruit Bats (Rousettus aegyptiacus) with Ebola Virus

PubMed Central

Paweska, Janusz T.; Storm, Nadia; Grobbelaar, Antoinette A.; Markotter, Wanda; Kemp, Alan; Jansen van Vuren, Petrus

2016-01-01

Colonized Egyptian fruit bats (Rousettus aegyptiacus), originating in South Africa, were inoculated subcutaneously with Ebola virus (EBOV). No overt signs of morbidity, mortality, or gross lesions were noted. Bats seroconverted by Day 10–16 post inoculation (p.i.), with the highest mean anti-EBOV IgG level on Day 28 p.i. EBOV RNA was detected in blood from one bat. In 16 other tissues tested, viral RNA distribution was limited and at very low levels. No seroconversion could be demonstrated in any of the control bats up to 28 days after in-contact exposure to subcutaneously-inoculated bats. The control bats were subsequently inoculated intraperitoneally, and intramuscularly with the same dose of EBOV. No mortality, morbidity or gross pathology was observed in these bats. Kinetics of immune response was similar to that in subcutaneously-inoculated bats. Viral RNA was more widely disseminated to multiple tissues and detectable in a higher proportion of individuals, but consistently at very low levels. Irrespective of the route of inoculation, no virus was isolated from tissues which tested positive for EBOV RNA. Viral RNA was not detected in oral, nasal, ocular, vaginal, penile and rectal swabs from any of the experimental groups. PMID:26805873

Effect of post-inoculation relative humidity on peanut infection by Sclerotinia minor

USDA-ARS?s Scientific Manuscript database

Stems of six-week-old plants of the cv Okrun (susceptible to Sclerotinia blight) were inoculated with S. minor. Two post-inoculation humidity regimes of 100% RH were used. In the first RH regime, one inoculation chamber was kept open for the duration of experiment (DOE), and five were closed for d...

Effect of microbial inoculants on the quality and aerobic stability of bermudagrass round-bale haylage.

PubMed

Arriola, K G; Queiroz, O C M; Romero, J J; Casper, D; Muniz, E; Hamie, J; Adesogan, A T

2015-01-01

The objective of this study was to compare the efficacy of using 4 commercially available microbial inoculants to improve the fermentation and aerobic stability of bermudagrass haylage. We hypothesized that the microbial inoculants would increase the fermentation and aerobic stability of the haylages. Bermudagrass (4-wk regrowth) was harvested and treated with (1) deionized water (control); (2) Buchneri 500 (B500; Lallemand Animal Nutrition, Milwaukee, WI) containing 1×10(5) of Pediococcus pentosaceus and 4×10(5) of Lactobacillus buchneri 40788; (3) Biotal Plus II (BPII; Lallemand Animal Nutrition) containing 1.2×10(5) of P. pentosaceus and Propionibacteria freudenreichii; (4) Silage Inoculant II (SI; AgriKing Inc., Fulton, IL) containing 1×10(5) of Lactobacillus plantarum and P. pentosaceus; and (5) Silo King (SK; AgriKing Inc.), containing 1×10(5) of L. plantarum, Enterococcus faecium, and P. pentosaceus, respectively. Forty round bales (8 per treatment; 441±26kg; 1.2×1.2 m diameter) were made and each was wrapped with 7 layers of plastic. Twenty bales were stored for 112 d and the remaining 20 were stored for 30 d and sampled by coring after intermediary storage periods of 0, 3, 7, and 30 d. The pH of control and inoculated haylages sampled on d 3 did not differ. However, B500 and BPII had lower pH (5.77±0.04 vs. 6.16±0.04; 5.06±0.13 vs. 5.52±0.13) than other treatments by d 7 and 30, respectively. At final bale opening on d 112, all treatments had lower pH than the control haylage (4.77±0.07 vs. 5.37±0.07). The B500, BPII, and SI haylages had greater lactic acid and lactic-to-acetic acid ratios than SK and control haylages. No differences were detected in neutral detergent fiber digestibility, dry matter losses, dry matter, lactic and acetic acid concentrations, and yeast and coliform counts. The SK haylage had lower clostridia counts compared with the control (1.19±0.23 vs. 1.99±0.23 cfu/g). Treatments B500, BPII, SI, and SK tended to reduce

Field Trials Reveal Ecotype-Specific Responses to Mycorrhizal Inoculation in Rice.

PubMed

Diedhiou, Abdala Gamby; Mbaye, Fatou Kine; Mbodj, Daouda; Faye, Mathieu Ndigue; Pignoly, Sarah; Ndoye, Ibrahima; Djaman, Koffi; Gaye, Souleymane; Kane, Aboubacry; Laplaze, Laurent; Manneh, Baboucarr; Champion, Antony

2016-01-01

The overuse of agricultural chemicals such as fertilizer and pesticides aimed at increasing crop yield results in environmental damage, particularly in the Sahelian zone where soils are fragile. Crop inoculation with beneficial soil microbes appears as a good alternative for reducing agricultural chemical needs, especially for small farmers. This, however, requires selecting optimal combinations of crop varieties and beneficial microbes tested in field conditions. In this study, we investigated the response of rice plants to inoculation with arbuscular mycorrhizal fungi (AMF) and plant growth promoting bacteria (PGPB) under screenhouse and field conditions in two consecutive seasons in Senegal. Evaluation of single and mixed inoculations with AMF and PGPB was conducted on rice (Oryza sativa) variety Sahel 202, on sterile soil under screenhouse conditions. We observed that inoculated plants, especially plants treated with AMF, grew taller, matured earlier and had higher grain yield than the non-inoculated plants. Mixed inoculation trials with two AMF strains were then conducted under irrigated field conditions with four O. sativa varieties, two O. glaberrima varieties and two interspecific NERICA varieties, belonging to 3 ecotypes (upland, irrigated, and rainfed lowland). We observed that the upland varieties had the best responses to inoculation, especially with regards to grain yield, harvest index and spikelet fertility. These results show the potential of using AMF to improve rice production with less chemical fertilizers and present new opportunities for the genetic improvement in rice to transfer the ability of forming beneficial rice-microbe associations into high yielding varieties in order to increase further rice yield potentials.

Field Trials Reveal Ecotype-Specific Responses to Mycorrhizal Inoculation in Rice

PubMed Central

Diedhiou, Abdala Gamby; Mbaye, Fatou Kine; Mbodj, Daouda; Faye, Mathieu Ndigue; Pignoly, Sarah; Ndoye, Ibrahima; Djaman, Koffi; Gaye, Souleymane; Kane, Aboubacry; Laplaze, Laurent; Manneh, Baboucarr; Champion, Antony

2016-01-01

The overuse of agricultural chemicals such as fertilizer and pesticides aimed at increasing crop yield results in environmental damage, particularly in the Sahelian zone where soils are fragile. Crop inoculation with beneficial soil microbes appears as a good alternative for reducing agricultural chemical needs, especially for small farmers. This, however, requires selecting optimal combinations of crop varieties and beneficial microbes tested in field conditions. In this study, we investigated the response of rice plants to inoculation with arbuscular mycorrhizal fungi (AMF) and plant growth promoting bacteria (PGPB) under screenhouse and field conditions in two consecutive seasons in Senegal. Evaluation of single and mixed inoculations with AMF and PGPB was conducted on rice (Oryza sativa) variety Sahel 202, on sterile soil under screenhouse conditions. We observed that inoculated plants, especially plants treated with AMF, grew taller, matured earlier and had higher grain yield than the non-inoculated plants. Mixed inoculation trials with two AMF strains were then conducted under irrigated field conditions with four O. sativa varieties, two O. glaberrima varieties and two interspecific NERICA varieties, belonging to 3 ecotypes (upland, irrigated, and rainfed lowland). We observed that the upland varieties had the best responses to inoculation, especially with regards to grain yield, harvest index and spikelet fertility. These results show the potential of using AMF to improve rice production with less chemical fertilizers and present new opportunities for the genetic improvement in rice to transfer the ability of forming beneficial rice-microbe associations into high yielding varieties in order to increase further rice yield potentials. PMID:27907023

Transformation with green fluorescent protein of Trichoderma harzianum 1051, a strain with biocontrol activity against Crinipellis perniciosa, the agent of witches'-broom disease of cocoa.

PubMed

Inglis, Peter W.; Queiroz, Paulo R.; Valadares-Inglis, M. Cléria

1999-04-01

A plasmid vector for fungal expression of an enhanced, red-shifted variant of the Aequoria victoriae green fluorescent protein was constructed by fusion of the EGFP gene to the highly expressed Aspergillus nidulans gpd promoter and the A. nidulans trpC terminator. This construction was introduced by cotransformation, using benomyl selection, into Trichoderma harzianum strain 1051, a strain being evaluated for the biological control of witches'-broom disease of cocoa caused by Crinipellis perniciosa. Epifluorescence microscopy was used to monitor germination and attachment of stable transformant conidia on the surface of C. perniciosa hyphae.

Inoculation stress hypothesis of environmental enrichment.

PubMed

Crofton, Elizabeth J; Zhang, Yafang; Green, Thomas A

2015-02-01

One hallmark of psychiatric conditions is the vast continuum of individual differences in susceptibility vs. resilience resulting from the interaction of genetic and environmental factors. The environmental enrichment paradigm is an animal model that is useful for studying a range of psychiatric conditions, including protective phenotypes in addiction and depression models. The major question is how environmental enrichment, a non-drug and non-surgical manipulation, can produce such robust individual differences in such a wide range of behaviors. This paper draws from a variety of published sources to outline a coherent hypothesis of inoculation stress as a factor producing the protective enrichment phenotypes. The basic tenet suggests that chronic mild stress from living in a complex environment and interacting non-aggressively with conspecifics can inoculate enriched rats against subsequent stressors and/or drugs of abuse. This paper reviews the enrichment phenotypes, mulls the fundamental nature of environmental enrichment vs. isolation, discusses the most appropriate control for environmental enrichment, and challenges the idea that cortisol/corticosterone equals stress. The intent of the inoculation stress hypothesis of environmental enrichment is to provide a scaffold with which to build testable hypotheses for the elucidation of the molecular mechanisms underlying these protective phenotypes and thus provide new therapeutic targets to treat psychiatric/neurological conditions. Copyright © 2014 Elsevier Ltd. All rights reserved.

Inoculation of Torulaspora delbrueckii as a bio-protection agent in winemaking.

PubMed

Simonin, Scott; Alexandre, Hervé; Nikolantonaki, Maria; Coelho, Christian; Tourdot-Maréchal, Raphaëlle

2018-05-01

In oenology, bio-protection consists in adding bacteria, yeasts or a mixture of microorganisms on grape must before fermentation in order to reduce the use of chemical compounds such as sulphites. More particularly, non-Saccharomyces yeasts are used as a total or partial alternative to sulphites. However, scientific data capable of proving the effectiveness of adding these yeasts on grape must is lacking. This study reports the analysis of antimicrobial and antioxidant effects of one non-Saccharomyces yeast, Torulaspora delbrueckii, inoculated at the beginning of the white winemaking process in two Burgundian wineries as an alternative to sulphiting. The implantation of the T. delbrueckii strain was successful in both wineries and had no impact on fermentation kinetics. Adding T. delbrueckii reduced biodiversity during the pre-fermentation stages compared to sulphited controls and it also effectively limited the development of spoilage microorganisms in the same way as the addition of sulphites. T. delbrueckii could protect must and wine from oxidation as demonstrated by the analysis of colour and phenolic compounds. This is the first evidence that early addition of T. delbrueckii during winemaking can be a microbiogical and chemical alternative to sulphites. However, its contribution seems to be matrix dependent. Copyright © 2018 Elsevier Ltd. All rights reserved.

Lactating cow response to lucerne silage inoculated with Lactobacillus plantarum

USDA-ARS?s Scientific Manuscript database

It is unclear why bacterial silage inoculants improve milk production in lactating dairy cattle. However, recent in vitro results suggest that inoculated silage effects on milk production may be tied to greater production of rumen microorganisms. Our objective was to determine if alfalfa silage trea...

[Queries related to the technology of soybean seed inoculation with Bradyrhizobium spp].

PubMed

Lodeiro, Aníbal R

2015-01-01

With the aim of exploiting symbiotic nitrogen fixation, soybean crops are inoculated with selected strains of Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens or Bradyrhizobium elkanii (collectively referred to as Bradyrhizobium spp.). The most common method of inoculation used is seed inoculation, whether performed immediately before sowing or using preinoculated seeds or pretreated seeds by the professional seed treatment. The methodology of inoculation should not only cover the seeds with living rhizobia, but must also optimize the chances of these rhizobia to infect the roots and nodulate. To this end, inoculated rhizobia must be in such an amount and condition that would allow them to overcome the competition exerted by the rhizobia of the allochthonous population of the soil, which are usually less effective for nitrogen fixation and thus dilute the effect of inoculation on yield. This optimization requires solving some queries related to the current knowledge of seed inoculation, which are addressed in this article. I conclude that the aspects that require further research are the adhesion and survival of rhizobia on seeds, the release of rhizobia once the seeds are deposited in the soil, and the movement of rhizobia from the vicinity of the seeds to the infection sites in the roots. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Influenza Virus Respiratory Infection and Transmission Following Ocular Inoculation in Ferrets

PubMed Central

Belser, Jessica A.; Gustin, Kortney M.; Maines, Taronna R.; Pantin-Jackwood, Mary J.; Katz, Jacqueline M.; Tumpey, Terrence M.

2012-01-01

While influenza viruses are a common respiratory pathogen, sporadic reports of conjunctivitis following human infection demonstrates the ability of this virus to cause disease outside of the respiratory tract. The ocular surface represents both a potential site of virus replication and a portal of entry for establishment of a respiratory infection. However, the properties which govern ocular tropism of influenza viruses, the mechanisms of virus spread from ocular to respiratory tissue, and the potential differences in respiratory disease initiated from different exposure routes are poorly understood. Here, we established a ferret model of ocular inoculation to explore the development of virus pathogenicity and transmissibility following influenza virus exposure by the ocular route. We found that multiple subtypes of human and avian influenza viruses mounted a productive virus infection in the upper respiratory tract of ferrets following ocular inoculation, and were additionally detected in ocular tissue during the acute phase of infection. H5N1 viruses maintained their ability for systemic spread and lethal infection following inoculation by the ocular route. Replication-independent deposition of virus inoculum from ocular to respiratory tissue was limited to the nares and upper trachea, unlike traditional intranasal inoculation which results in virus deposition in both upper and lower respiratory tract tissues. Despite high titers of replicating transmissible seasonal viruses in the upper respiratory tract of ferrets inoculated by the ocular route, virus transmissibility to naïve contacts by respiratory droplets was reduced following ocular inoculation. These data improve our understanding of the mechanisms of virus spread following ocular exposure and highlight differences in the establishment of respiratory disease and virus transmissibility following use of different inoculation volumes and routes. PMID:22396651

Association of protein tyrosine phosphatase, non-receptor type 22 +1858C→T polymorphism and susceptibility to vitiligo: Systematic review and meta-analysis.

PubMed

Agarwal, Silky; Changotra, Harish

2017-01-01

Protein tyrosine phosphatase, non-receptor type 22 gene, which translates to lymphoid tyrosine phosphatase, is considered to be a susceptibility gene marker associated with several autoimmune diseases. Several studies have demonstrated the association of protein tyrosine phosphatase, non-receptor type 22 +1858C→T polymorphism with vitiligo. However, these studies showed conflicting results. Meta-analysis of the same was conducted earlier that included fewer number of publications in their study. We performed a meta-analysis of a total of seven studies consisting of 2094 cases and 3613 controls to evaluate the possible association of protein tyrosine phosphatase, non-receptor type 22 +1858C>T polymorphism with vitiligo susceptibility. We conducted a literature search in PubMed, Google Scholar and Dogpile for all published paper on protein tyrosine phosphatase, non-receptor type 22 +1858C→T polymorphism and vitiligo risk till June 2016. Data analysis was performed by RevMan 5.3 and comprehensive meta-analysis v3.0 software. Meta-analysis showed an overall significant association of protein tyrosine phosphatase, non- receptor type 22 +1858C→T polymorphism with vitiligo in all models (allelic model [T vs. C]: odds ratio = 1.50, 95% confidence interval [1.32-1.71], P< 0.001; dominant model [TT + CT vs. CC]: odds ratio = 1.61, 95% confidence interval [1.16-2.24], P = 0.004; recessive model [TT vs. CT + CC]: odds ratio = 4.82, 95% confidence interval [1.11-20.92], P = 0.04; homozygous model [TT vs. CC]: odds ratio = 5.34, 95% confidence interval [1.23-23.24], P = 0.03; co-dominant model [CT vs. CC]: odds ratio = 1.52, 95% confidence interval [1.09-2.13], P = 0.01). No publication bias was detected in the funnel plot study. Limited ethnic-based studies, unable to satisfy data by gender or vitiligo-type are some limitations of the present meta-analysis. Stratifying data by ethnicity showed an association of protein tyrosine phosphatase, non-receptor type 22 +1858C→T

Expression of retinoid-related orphan receptor (ROR)γt on NK22 cells in the peripheral blood and uterine endometrium of women with unexplained recurrent pregnancy loss and unexplained infertility.

PubMed

Fuchinoue, Kohei; Fukui, Atsushi; Chiba, Hitomi; Kamoi, Mai; Funamizu, Ayano; Taima, Ayako; Fukuhara, Rie; Mizunuma, Hideki

2016-11-01

Recently, NK22 cells, a subset of interleukin (IL)-22-producing natural killer (NK) cells, were identified. We have previously reported the higher percentage of NK22 cells in women suffering recurrent pregnancy loss (RPL). Moreover, we have also reported lower expression of NKp46, a kind of natural cytotoxicity receptor (NCR), on NK cells and the changes of NK cell producing cytokines in women who experience RPL. NK22 cells express NCRs, such as NKp44 or NKp46. Retinoid-related orphan receptor γt (RORγt) is known as a regulator of NK22 cells; however, in NK22 cells of peripheral blood (PB) and the uterine endometrium (UE), the relationship between NCRs and RORγt is unclear. We investigate RORγt expression NK22 cells in the PB and UE of women with unexplained infertility (uI) or unexplained RPL (uRPL). Lymphocytes were extracted from PB and UE, derived from women with uI or uRPL. Expression of RORγt and NCRs in NK cells and NK cell-produced cytokines were analyzed by flow cytometry. CD56 + /NKp46 + /RORγt + cells were positively correlated with CD56 + /IL-22 + cells in both PB and UE. CD56 bright /NKp46 bright /RORγt + cells were significantly higher in uRPL than in uI, and endometrial CD56 bright /NKp46 bright /RORγt + cells were positively correlated with PB. In UE, CD56 bright /RORγt + cells were negatively correlated with CD56 bright /interferon-γ + and CD56 bright /tumor necrosis factor-α + cells of uRPL. RORγt may be associated with NK22 cells in reproduction. Particularly, higher expression of RORγt may be associated with elevated NK22 cells in uRPL. © 2016 Japan Society of Obstetrics and Gynecology.

The effects of arbuscular mycorrhizal fungal inoculation at a roadside prairie restoration site.

PubMed

White, Jennifer A; Tallaksen, J; Charvat, I

2008-01-01

Arbuscular mycorrhizal fungi (AMF) may play an important role in ecological succession, but few studies have documented the effectiveness of mycorrhizal inoculation at restoration/reclamation sites. At a roadside prairie restoration in Shakopee, Minnesota, we compared AMF root colonization and resulting vegetative cover among four inoculation treatments. After 15 mo of growth, we found that AMF colonization was high in all treatments but was significantly higher in treatments that received AMF inoculum propagated from a local prairie site or commercially available inoculum than the uninoculated control. For the prairie inoculum, this increase in colonization occurred whether the inoculum was applied with seeds in furrows or broadcast with seeds on the soil surface. However, increased colonization did not discernibly affect the restored vegetation; neither total vegetative cover nor the proportion "desired" prairie vegetation differed among inoculation treatments. By the end of the third growing season (27 mo after planting) there were no longer differences in AMF colonization among the inoculation treatments nor were there differences in vegetative cover. It is likely that natural recolonization of the plots by remnant AMF populations at the site limited the duration of the inoculation effect. This natural recolonization, in combination with relatively high soil phosphorus levels, likely rendered inoculation unnecessary. In contrast to previous published studies of AMF inoculation in landscape restorations, this study shows that AMF inoculation may not be warranted under some circumstances.

Inoculation density is affecting growth conditions of Listeria monocytogenes on fresh cut lettuce.

PubMed

McManamon, Oisin; Scollard, Johann; Schmalenberger, Achim

2017-11-27

Listeria monocytogenes is a particular risk for the ready-to-eat food sector because of its ability to grow in various environmental conditions. In the literature, growth and survival of L. monocytogenes on food is tested using inoculation densities ranging from less than 10 2 to over 10 5  CFU g -1 . Inoculation densities on food have been rarely tested as a factor for growth. In this study, inoculation densities from 10 2 to 10 5 of L. monocytogenes were tested on iceberg lettuce (Lactuca sativa) in modified atmospheres and air in model packages at 4 and 8 °C to identify any potential inoculation density effects. On days 0, 2, 5 and 7, L. monocytogenes was extracted from the lettuce surface and enumerated via selective media. The resulting growth curves identified a significant inoculation density effect at 4 and 8 °C with significantly higher amounts of growth (1-2 logs) when lettuce was inoculated at 10 2  CFU g -1 as opposed to 10 4 and 10 5  CFU g -1 . In contrast, the use of different atmospheres had limited influence on growth of L. monocytogenes. In conclusion, greater emphasis on inoculation density of L. monocytogenes should be taken in inoculation experiments when confirmation of growth or the efficacies of growth inhibiting treatments are tested on ready-to-eat food such as lettuce.

Re-Thinking Anxiety: Using Inoculation Messages to Reduce and Reinterpret Public Speaking Fears.

PubMed

Jackson, Ben; Compton, Josh; Thornton, Ashleigh L; Dimmock, James A

2017-01-01

Inoculation theory offers a framework for protecting individuals against challenges to an existing attitude, belief, or state. Despite the prevalence and damaging effects of public speaking anxiety, inoculation strategies have yet to be used to help individuals remain calm before and during public speaking. We aimed to test the effectiveness of an inoculation message for reducing the onset of public speaking anxiety, and helping presenters interpret their speech-related anxiety more positively. Participants (Mage = 20.14, SD = 2.72) received either an inoculation (n = 102) or control (n = 128) message prior to engaging a public speaking task and reported a range of anxiety-related perceptions. Accounting for personality characteristics and perceptions of task importance, and relative to control participants, those who received the inoculation message reported significantly lower pre-task anxiety, and following the task, reported that they had experienced lower somatic anxiety, and that the inoculation message had caused them to view their nerves in a less debilitating light. Inoculation messages may be an effective strategy for helping participants reframe and reduce their apprehension about public speaking, and investigating their efficacy in other stress-inducing contexts may be worthwhile.

Re-Thinking Anxiety: Using Inoculation Messages to Reduce and Reinterpret Public Speaking Fears

PubMed Central

Jackson, Ben; Compton, Josh; Thornton, Ashleigh L.; Dimmock, James A.

2017-01-01

Inoculation theory offers a framework for protecting individuals against challenges to an existing attitude, belief, or state. Despite the prevalence and damaging effects of public speaking anxiety, inoculation strategies have yet to be used to help individuals remain calm before and during public speaking. We aimed to test the effectiveness of an inoculation message for reducing the onset of public speaking anxiety, and helping presenters interpret their speech-related anxiety more positively. Participants (Mage = 20.14, SD = 2.72) received either an inoculation (n = 102) or control (n = 128) message prior to engaging a public speaking task and reported a range of anxiety-related perceptions. Accounting for personality characteristics and perceptions of task importance, and relative to control participants, those who received the inoculation message reported significantly lower pre-task anxiety, and following the task, reported that they had experienced lower somatic anxiety, and that the inoculation message had caused them to view their nerves in a less debilitating light. Inoculation messages may be an effective strategy for helping participants reframe and reduce their apprehension about public speaking, and investigating their efficacy in other stress-inducing contexts may be worthwhile. PMID:28125618

Enhancing immune responses to inactivated porcine parvovirus oil emulsion vaccine by co-inoculating porcine transfer factor in mice.

PubMed

Wang, Rui-ning; Wang, Ya-bin; Geng, Jing-wei; Guo, Dong-hui; Liu, Fang; Chen, Hong-ying; Zhang, Hong-ying; Cui, Bao-an; Wei, Zhan-yong

2012-07-27

Inactivated porcine parvovirus (PPV) vaccines are available commercially and widely used in the breeding herds. However, inactivated PPV vaccines have deficiencies in induction of specific cellular immune response. Transfer factor (TF) is a material that obtained from the leukocytes, and is a novel immune-stimulatory reagent that as a modulator of the immune system. In this study, the immunogenicity of PPV oil emulsion vaccine and the immuno-regulatory activities of TF were investigated. The inactivated PPV oil emulsion vaccines with or without TF were inoculated into BALB/c mice by subcutaneous injection. Then humoral and cellular immune responses were evaluated by indirect enzyme-linked immunosorbent assays (ELISA), fluorescence-activated cell sorter analyses (FACS). The results showed that the PPV specific immune responses could be evoked in mice by inoculating with PPV oil emulsion vaccine alone or by co-inoculation with TF. The cellular immune response levels in the co-inoculation groups were higher than those groups receiving the PPV oil emulsion vaccine alone, with the phenomena of higher level of IFN-γ, a little IL-6 and a trace of IL-4 in serum, and a vigorous T-cell response. However, there was no significant difference in antibody titers between TF synergy inactivated vaccine and the inactivated vaccine group (P>0.05). In conclusion, these results suggest that TF possess better cellular immune-enhancing capability and would be exploited into an effective immune-adjuvant for inactivated vaccines. Copyright © 2012 Elsevier Ltd. All rights reserved.

Inoculation of female American black bears (Ursus americanus) with partially purified porcine zona pellucidae limits cub production.

PubMed

Lane, V M; Liu, I K M; Casey, K; vanLeeuwen, E M G; Flanagan, D R; Murata, K; Munro, C

2007-01-01

The present 2-year study investigated the feasibility of using porcine zona pellucidae (pZP) as antigen for immunocontraception in American black bears. Sows, 3-6 years of age, were administered either two doses of 250 microg pZP with Freund's adjuvant (n = 10) or adjuvant alone (n = 5), one in April and one in May, and were kept away from the boars until June. Serum samples were collected before injections and before denning (November). The presence of sows with cubs at side was observed during premature emergence from denning. First-year results indicated that anti-pZP antibody titres in vaccinated sows were 2.5-9.0-fold (range) higher compared with non-vaccinated sows and that the vaccinated sows were threefold less likely to become pregnant (P = 0.167). Control and vaccinated bears produced 1.6 and 0.2 cubs per sow, respectively (P = 0.06). The second-year study investigated the feasibility of using pZP sequestered in a controlled-release pellet and a water-soluble adjuvant (QS-21) to avoid regulatory problems associated with Freund's adjuvant. Sows in the treatment group (n = 22) were administered a single dose of an emulsion of 250 microg pZP and 150 microg QS-21 plus a pellet containing 70-90 microg pZP for delayed release as booster dose. Control sows (n = 5) received the QS-21 adjuvant in pellet alone. Serum samples were collected before inoculations (April) and before denning (November). Seven cubs were born to the five control sows, but none was born to the 22 vaccinated sows (P < 0.001). Anti-pZP antibody mean absorbance ratios in control sows remained at background levels, whereas vaccinated sows had ratios fourfold higher than controls. Two-dimensional polyacrylamide gel electrophoresis and immunohistochemical localisation confirmed immunoreactivity of sera from inoculated bears. We conclude that cub production in the American black bear can be effectively limited with either two injections of 250 microg pZP or a single inoculation of partially

Efficient Recreation of t(11;22) EWSR1-FLI1+ in Human Stem Cells Using CRISPR/Cas9.

PubMed

Torres-Ruiz, Raul; Martinez-Lage, Marta; Martin, Maria C; Garcia, Aida; Bueno, Clara; Castaño, Julio; Ramirez, Juan C; Menendez, Pablo; Cigudosa, Juan C; Rodriguez-Perales, Sandra

2017-05-09

Efficient methodologies for recreating cancer-associated chromosome translocations are in high demand as tools for investigating how such events initiate cancer. The CRISPR/Cas9 system has been used to reconstruct the genetics of these complex rearrangements at native loci while maintaining the architecture and regulatory elements. However, the CRISPR system remains inefficient in human stem cells. Here, we compared three strategies aimed at enhancing the efficiency of the CRISPR-mediated t(11;22) translocation in human stem cells, including mesenchymal and induced pluripotent stem cells: (1) using end-joining DNA processing factors involved in repair mechanisms, or (2) ssODNs to guide the ligation of the double-strand break ends generated by CRISPR/Cas9; and (3) all-in-one plasmid or ribonucleoprotein complex-based approaches. We report that the generation of targeted t(11;22) is significantly increased by using a combination of ribonucleoprotein complexes and ssODNs. The CRISPR/Cas9-mediated generation of targeted t(11;22) in human stem cells opens up new avenues in modeling Ewing sarcoma. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Effect of inoculum size, bacterial species, type of surfaces and contact time to the transfer of foodborne pathogens from inoculated to non-inoculated beef fillets via food processing surfaces.

PubMed

Gkana, E; Chorianopoulos, N; Grounta, A; Koutsoumanis, K; Nychas, G-J E

2017-04-01

The objective of the present study was to determine the factors affecting the transfer of foodborne pathogens from inoculated beef fillets to non-inoculated ones, through food processing surfaces. Three different levels of inoculation of beef fillets surface were prepared: a high one of approximately 10 7  CFU/cm 2 , a medium one of 10 5  CFU/cm 2 and a low one of 10 3  CFU/cm 2 , using mixed-strains of Listeria monocytogenes, or Salmonella enterica Typhimurium, or Escherichia coli O157:H7. The inoculated fillets were then placed on 3 different types of surfaces (stainless steel-SS, polyethylene-PE and wood-WD), for 1 or 15 min. Subsequently, these fillets were removed from the cutting boards and six sequential non-inoculated fillets were placed on the same surfaces for the same period of time. All non-inoculated fillets were contaminated with a progressive reduction trend of each pathogen's population level from the inoculated fillets to the sixth non-inoculated ones that got in contact with the surfaces, and regardless the initial inoculum, a reduction of approximately 2 log CFU/g between inoculated and 1st non-inoculated fillet was observed. S. Typhimurium was transferred at lower mean population (2.39 log CFU/g) to contaminated fillets than E. coli O157:H7 (2.93 log CFU/g), followed by L. monocytogenes (3.12 log CFU/g; P < 0.05). Wooden surfaces (2.77 log CFU/g) enhanced the transfer of bacteria to subsequent fillets compared to other materials (2.66 log CFU/g for SS and PE; P < 0.05). Cross-contamination between meat and surfaces is a multifactorial process strongly depended on the species, initial contamination level, kind of surface, contact time and the number of subsequent fillet, according to analysis of variance. Thus, quantifying the cross-contamination risk associated with various steps of meat processing and food establishments or households can provide a scientific basis for risk management of such products. Copyright © 2016 Elsevier Ltd

Effect of post-inoculation relative humidity (RH) on peanut infection by Sclerotinia sclerotiorum

USDA-ARS?s Scientific Manuscript database

Stems of six-week-old plants of the cv Okrun (susceptible to Sclerotinia blight) were inoculated with S. sclerotiorum, isolated from pumpkin. Two post-inoculation humidity regimes of 100% RH were used. In the first RH regime, one inoculation chamber was kept open for the duration of experiment (DO...

IL-22 promotes Fas expression in oligodendrocytes and inhibits FOXP3 expression in T cells by activating the NF-κB pathway in multiple sclerosis.

PubMed

Zhen, Jin; Yuan, Jun; Fu, Yongwang; Zhu, Runxiu; Wang, Meiling; Chang, Hong; Zhao, Yan; Wang, Dong; Lu, Zuneng

2017-02-01

Multiple sclerosis (MS) is characterized by an increase in interleukin-22 and Fas, and a decrease in FOXP3, among other factors. In this study, we examined patients with MS and healthy control subjects and used the experimental autoimmune encephalomyelitis (EAE) animal model to identify the effects of IL-22 on oligodendrocytes and T cells in MS development. In MS, the expression of Fas in oligodendrocytes and IL-22 in CD4 + CCR4 + CCR6 + CCR10 + T cells was enhanced. Ikaros and FOXP3 were both decreased in T cells. Depending on exogenous IL-22, Fas increased the phosphorylation of mitogen- and stress-activated protein kinase 1 and activated the nuclear factor-κB pathway in oligodendrocytes, leading to an increase in Fas and oligodendrocyte apoptosis. IL-22 decreased FOXP3 expression by activating NF-κB, and it further inhibited PTEN and Ikaros expression. Tregs reversed the functions of IL-22. Taken together, these findings help to elucidate the mechanisms of IL-22 in MS development. Copyright © 2016 Elsevier Ltd. All rights reserved.

Potential woodpecker nest trees through artificial inoculation of heart rots

Treesearch

Richard N. Conner; James G. Dickson; J. Howard Williamson

1983-01-01

We suggest that the fungus Spongipellis pachyodon might be used to artificially create suitable hardwood nest trees for woodpeckers in both young and older trees and when supplies of potential nest trees are limited. Sizes of trees suitable for inoculation, inoculation heights, and densities of snags are suggested for six species of woodpeckers.

Effects of water stress, organic amendment and mycorrhizal inoculation on soil microbial community structure and activity during the establishment of two heavy metal-tolerant native plant species.

PubMed

Fernández, D A; Roldán, A; Azcón, R; Caravaca, F; Bååth, E

2012-05-01

Our aim was to examine the effect of water stress on plant growth and development of two native plant species (Tetraclinis articulata and Crithmum maritimum) and on microbial community composition and activity in the rhizosphere soil, following the addition of an organic amendment, namely sugar beet residue (SBR), and/or the inoculation with an arbuscular mycorrhizal (AM) fungus, namely Glomus mosseae, in a non-sterile heavy metal-polluted soil. The AM inoculation did not have any significant effect on plant growth of both species. In T. articulata, SBR increased shoot growth, foliar P, total phospholipid fatty acids (PLFA), fungi-related PLFA, AM fungi-related neutral lipid fatty acid, bacterial gram-positive/gram-negative PLFA ratio and the β-glucosidase and dehydrogenase activities. SBR and AM inoculation increased phosphatase activity in T. articulata plants grown under drought conditions. In both plants, there was a synergistic effect between AM inoculation and SBR on mycorrhizal colonisation under drought conditions. In C. maritimum, the increase produced by the SBR on total amounts of PLFA, bacterial gram-positive-related PLFA and bacterial gram-negative-related PLFA was considerably higher under drought conditions. Our results suggest that the effectiveness of the amendment with regard to stimulating microbial communities and plant growth was largely limited by drought, particularly for plant species with a low degree of mycorrhizal colonisation.

Rapid Dissemination of SIV Follows Multisite Entry after Rectal Inoculation

PubMed Central

Prétet, Jean-Luc; Michel-Salzat, Alice; Messent, Valérie; Bogdanova, Anna; Couëdel-Courteille, Anne; Souil, Evelyne; Cheynier, Rémi; Butor, Cécile

2011-01-01

Receptive ano-rectal intercourse is a major cause of HIV infection in men having sex with men and in heterosexuals. Current knowledge of the mechanisms of entry and dissemination during HIV rectal transmission is scarce and does not allow the development of preventive strategies. We investigated the early steps of rectal infection in rhesus macaques inoculated with the pathogenic isolate SIVmac251 and necropsied four hours to nine days later. All macaques were positive for SIV. Control macaques inoculated with heat-inactivated virus were consistently negative for SIV. SIV DNA was detected in the rectum as early as four hours post infection by nested PCR for gag in many laser-microdissected samples of lymphoid aggregates and lamina propria but never in follicle-associated epithelium. Scarce SIV antigen positive cells were observed by immunohistofluorescence in the rectum, among intraepithelial and lamina propria cells as well as in clusters in lymphoid aggregates, four hours post infection and onwards. These cells were T cells and non-T cells that were not epithelial cells, CD68+ macrophages, DC-SIGN+ cells or fascin+ dendritic cells. DC-SIGN+ cells carried infectious virus. Detection of Env singly spliced mRNA in the mucosa by nested RT-PCR indicated ongoing viral replication. Strikingly, four hours post infection colic lymph nodes were also infected in all macaques as either SIV DNA or infectious virus was recovered. Rapid SIV entry and dissemination is consistent with trans-epithelial transport. Virions appear to cross the follicle-associated epithelium, and also the digestive epithelium. Viral replication could however be more efficient in lymphoid aggregates. The initial sequence of events differs from both vaginal and oral infections, which implies that prevention strategies for rectal transmission will have to be specific. Microbicides will need to protect both digestive and follicle-associated epithelia. Vaccines will need to induce immunity in lymph nodes

Early detection of tumor cells by innate immune cells leads to T(reg) recruitment through CCL22 production by tumor cells.

PubMed

Faget, Julien; Biota, Cathy; Bachelot, Thomas; Gobert, Michael; Treilleux, Isabelle; Goutagny, Nadège; Durand, Isabelle; Léon-Goddard, Sophie; Blay, Jean Yves; Caux, Christophe; Ménétrier-Caux, Christine

2011-10-01

In breast carcinomas, patient survival seems to be negatively affected by the recruitment of regulatory T cells (T(reg)) within lymphoid aggregates by CCL22. However, the mechanisms underpinning this process, which may be of broader significance in solid tumors, have yet to be described. In this study, we determined how CCL22 production is controlled in tumor cells. In human breast carcinoma cell lines, CCL22 was secreted at low basal levels that were strongly increased in response to inflammatory signals [TNF-α, IFN-γ, and interleukin (IL)-1β], contrasting with CCL17. Primary breast tumors and CD45(+) infiltrating immune cells appeared to cooperate in driving CCL22 secretion, as shown clearly in cocultures of breast tumor cell lines and peripheral blood mononuclear cells (PBMC) or their supernatants. We determined that monocyte-derived IL-1β and TNF-α are key players as monocyte depletion or neutralization of these cytokines attenuated secretion of CCL22. However, when purified monocytes were used, exogenous human IFN-γ was also required to generate this response suggesting a role for IFN-γ-producing cells within PBMCs. In this setting, we found that human IFN-γ could be replaced by the addition of (i) IL-2 or K562-activated natural killer (NK) cells or (ii) resting NK cells in the presence of anti-MHC class I antibody. Taken together, our results show a dialogue between NK and tumor cells leading to IFN-γ secretion, which in turn associates with monocyte-derived IL-1β and TNF-α to drive production of CCL22 by tumor cells and subsequent recruitment of T(reg). As one validation of this conclusion in primary breast tumors, we showed that NK cells and macrophages tend to colocalize within tumors. In summary, our findings suggest that at early times during tumorigenesis, the detection of tumor cells by innate effectors (monocytes and NK cells) imposes a selection for CCL22 secretion that recruits T(reg) to evade this early antitumor immune response.

Seasonal trends in response to inoculation of coast live oak with Phytophthora ramorum

Treesearch

Richard S. Dodd; Daniel Hüberli; Tamar Y. Harnik; Brenda O' Dell; Matteo Garbelotto

2006-01-01

We developed a branch cutting inoculation method to provide a controlled system for studying variation in response to inoculation of coast live oak (Quercus agrifolia) with Phytophthora ramorum. This method has advantages over inoculations of trees in the field, in containing the inoculum and in allowing high levels of replication...

Effects of inoculation of commercial starter cultures on the quality and histamine accumulation in fermented sausages.

PubMed

Wang, Xinhui; Ren, Hongyang; Wang, Wei; Zhang, Yin; Bai, Ting; Li, Junxia; Zhu, Wenyou

2015-02-01

To meet the requirements of high-quality safe products, starter cultures are used to produce fermented sausages. The effects of 3 commercial starter cultures, namely SM-194, T-SPX, and SM-181, on histamine accumulation and quality parameters including microbial quality, pH, water activity, and total volatile base nitrogen, as well as the color and texture properties, were evaluated during the fermentation and ripening of fermented sausages. Although initial counts of Escherichia coli, Enterobacteriaceae, and Pseudomonas were similar in the 4 batches, the growth of these microorganisms was significantly inhibited (P < 0.05) in batches SM-194, T-SPX, and SM-181 throughout the fermentation and ripening period. The counts of E. coli, Enterobacteriaceae, and Pseudomonas increased to maximum levels of 3.89, 4.41, and 5.15 log10 colony forming units/g in the control sausages, respectively. At the end of ripening, the levels of histamine were 8.85, 0.32, 7.82, and 3.18 mg/kg for batches C, SM-194, T-SPX, and SM-181, respectively. The results revealed that commercial starter cultures, particularly starter cultures SM-194 and SM-181, made a great contribution to histamine reduction. In addition, batches inoculated with starter cultures showed a stronger acidification and lower level of total volatile base nitrogen than the control sample during production (P < 0.05). In conclusion, it seems that the inoculation of commercial starter cultures, particularly starter cultures SM-194 and SM-181, contributes to improving microbial quality, hygienic quality and food safety of fermented sausages. © 2015 Institute of Food Technologists®

[Rhizosphere microbial impacts of alleviating faba bean Fusarium wilt with inoculating AM fungi].

PubMed

Dong, Yan; Dong, Kun; Yang, Zhi Xian; Tang, Li; Zheng, Yi

2016-12-01

Greenhouse pot trials were conducted to investigate the effects of arbuscular mycorrhizal fungus (Glomus mosseae, Glomus tortuosum, Glomus intraradices and Glomus etunicatum) inoculation on the seedling growth, occurance of Fusarium wilt, population of Fusarium oxysporum and rhizosphere microbial community functional diversity in faba bean rhizosphere soil. Results showed that after inoculation of G. mosseae, G. tortuosum, G. intraradices and G. etunicatum, the shoot and root fresh mass of faba bean seedlings increased significantly, the disease index of faba bean fusarium wilt decreased significantly by 94.0%, 60.0%, 64.0% and 94.0%, respectively, the amount of F. oxysporum of faba bean rhizosphere decreased significantly by 98.6%, 74.3%, 77.8% and 90.4%, respectively. The best inhibitory effects to Fusarium wilt were with G. mosseae and G. etunicatum treatments. Inoculation of G. mosseae, G. tortuosum and G. etunicatum significantly increased carbon sources utilization ability of carbohydrates, amino acids, carboxylic acids and phenolic acids, with the average well color development (AWCD) value being increased by 34.4%, 31.5% and 50.8% respectively, but such significant differences were not observed with inoculation of G. intraradice. Principal component analyses showed that inoculation of G. mosseae, G. tortuosum and G. etunicatum fungi changed the rhizospheric microbial community composition. Correlation analyses showed that the utilization of carbohydrates carbon sources (β-Methyl-D-glucoside, D-Galacturonic acid, D-Mannitol, N-Acetyl-D-Glucosamine, D-Cellobiose,) and carboxylic acids carbon sources (D-Galactonic acid-γ-Lactone) were significantly increased after inoculation of G. tortuosum, and the utilization of L-Arginine and 4-Hydroxy benzoic acid significantly increased after inoculation of G. mosseae and G. etunicatum. Carbohydrates, carboxylic acids were main carbon sources utilized by rhizosphere microbes after G. tortuosum and G. intraradices

Serological responses in chimpanzees inoculated with human immunodeficiency virus glycoprotein (gp120) subunit vaccine

DOE Office of Scientific and Technical Information (OSTI.GOV)

Arthur, L.O.; Pyle, S.W.; Nara, P.L.

1987-12-01

The major envelope glycoprotein of a human immunodeficiency virus (HIV) has been purified and was utilized as a prototype vaccine in chimpanzees. The 120,000-dalton glycoprotein (gp120) was purified from membranes of human T-lymphotropic virus (HTLV)-IIIB-infected cells and the final preparation contained low levels to no detectable HTLV-IIIB core antigen (p24) and low levels of endotoxin. Chimpanzees inoculated with gp120 responded by developing antibodies that precipitated radiolabeled gp120 and neutralized in vitro infection of HTLV-IIIB. Antibodies to HTLV-IIIB p24 were not detected in the gp120-immunized chimpanzees. Peripheral blood leukocytes from the vaccinated animals were examined for T4/sup +/ and T8/sup +/more » cells, and no decrease in the T4/T8 ratio was found, indicating that immunization with a ligand (gp120) that binds to T4 has not detectable adverse effect on the population of T4/sup +/ cells. The only current animal model that can be reproducibly infected with HIV is the chimpanzee. Immunization of chimpanzees with HIV proteins will provide an experimental system for testing the effectiveness of prototype vaccines for preventing HIV infection in vivo.« less

Striatal dopamine D2/3 receptor regulation by stress inoculation in squirrel monkeys.

PubMed

Lee, Alex G; Nechvatal, Jordan M; Shen, Bin; Buckmaster, Christine L; Levy, Michael J; Chin, Frederick T; Schatzberg, Alan F; Lyons, David M

2016-06-01

Intermittent mildly stressful situations provide opportunities to learn, practice, and improve coping in a process called stress inoculation. Stress inoculation also enhances cognitive control and response inhibition of impulsive motivated behavior. Cognitive control and motivation have been linked to striatal dopamine D2 and/or D3 receptors (DRD2/3) in rodents, monkeys, and humans. Here, we study squirrel monkeys randomized early in life to stress inoculation with or without maternal companionship and a no-stress control treatment condition. Striatal DRD2/3 availability in adulthood was measured in vivo by [ 11 C]raclopride binding using positron emission tomography (PET). DRD2/3 availability was greater in caudate and putamen compared to ventral striatum as reported in PET studies of humans and other non-human primates. DRD2/3 availability in ventral striatum was also consistently greater in stress inoculated squirrel monkeys compared to no-stress controls. Squirrel monkeys exposed to stress inoculation in the presence of their mother did not differ from squirrel monkeys exposed to stress inoculation without maternal companionship. Similar effects in different social contexts extend the generality of our findings and together suggest that stress inoculation increases striatal DRD2/3 availability as a correlate of cognitive control in squirrel monkeys.

Characterization of Malaysian Trichoderma isolates using random amplified microsatellites (RAMS).

PubMed

Siddiquee, Shafiquzzaman; Tan, Soon Guan; Yusuf, Umi Kalsom; Fatihah, Nur Hasan Nudin; Hasan, Md Mainul

2012-01-01

Trichoderma species are commercially applied as biocontrol agents against numerous plant pathogenic fungi due to their production of antifungal metabolites, competition for nutrients and space, and mycoparasitism. However, currently the identification of Trichoderma species from throughout the world based on micro-morphological descriptions is tedious and prone to error. The correct identification of Trichoderma species is important as several traits are species-specific. The Random Amplified Microsatellites (RAMS) analysis done using five primers in this study showed different degrees of the genetic similarity among 42 isolates of this genus. The genetic similarity values were found to be in the range of 12.50-85.11% based on a total of 76 bands scored in the Trichoderma isolates. Of these 76 bands, 96.05% were polymorphic, 3.95% were monomorphic and 16% were exclusive bands. Two bands (250 bp and 200 bp) produced by primer LR-5 and one band (250 bp) by primer P1A were present in all the Trichoderma isolates collected from healthy and infected oil palm plantation soils. Cluster analysis based on UPGMA of the RAMS marker data showed that T. harzianum, T. virens and T. longibrachiatum isolates were grouped into different clades and lineages. In this study we found that although T. aureoviride isolates were morphologically different when compared to T. harzianum isolates, the UPGMA cluster analysis showed that the majority isolates of T. aureoviride (seven from nine) were closely related to the isolates of T. harzianum.

IL13Rα2 expression identifies tissue-resident IL-22 producing PLZF+ innate T cells in the human liver.

PubMed

Paquin-Proulx, Dominic; Greenspun, Benjamin C; Pasquet, Lise; Strunz, Benedikt; Aleman, Soo; Falconer, Karolin; Terabe, Masaki; Berzofsky, Jay A; Sandberg, Johan K; Melum, Espen; Nixon, Douglas F; Björkström, Niklas K

2018-04-20

Innate lymphocytes are selectively enriched in the liver where they have important roles in liver immunology. Murine studies have shown that type I NKT cells can promote liver inflammation whereas type II NKT cells have an anti-inflammatory role. In humans, type II NKT cells were found to accumulate in the gut during inflammation and IL13Rα2 was proposed as a marker for these cells. In the human liver, less is known about type I and II NKT cells. Here, we studied the phenotype and function of human liver T cells expressing IL13Rα2. We found that IL13Rα2 was expressed by around 1% of liver resident memory T cells but not on circulating T cells. In support of their innate-like T cell character, the IL13Rα2 + T cells had higher expression of PLZF compared to IL13Rα2 - T cells and possessed the capacity to produce IL-22. However, only a minority of human liver sulfatide-reactive type II NKT cells expressed IL13Rα2. Collectively, these findings suggest that IL13Rα2 identifies tissue-resident intrahepatic T cells with innate characteristics and the capacity to produce IL-22. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Mathematical modeling of enzyme production using Trichoderma harzianum P49P11 and sugarcane bagasse as carbon source.

PubMed

Gelain, Lucas; da Cruz Pradella, José Geraldo; da Costa, Aline Carvalho

2015-12-01

A mathematical model to describe the kinetics of enzyme production by the filamentous fungus Trichoderma harzianum P49P11 was developed using a low cost substrate as main carbon source (pretreated sugarcane bagasse). The model describes the cell growth, variation of substrate concentration and production of three kinds of enzymes (cellulases, beta-glucosidase and xylanase) in different sugarcane bagasse concentrations (5; 10; 20; 30; 40 gL(-1)). The 10 gL(-1) concentration was used to validate the model and the other to parameter estimation. The model for enzyme production has terms implicitly representing induction and repression. Substrate variation was represented by a simple degradation rate. The models seem to represent well the kinetics with a good fit for the majority of the assays. Validation results indicate that the models are adequate to represent the kinetics for a biotechnological process. Copyright © 2015 Elsevier Ltd. All rights reserved.

Ectomycorrhizal inoculation of containerized Western conifer seedlings.

Treesearch

Randy. Molina

1980-01-01

Of 15 ectomycorrhizal fungi inoculated onto five container-grown conifer species (Larix occidentalis, Pinus contorta, P. ponderosa, Pseudotsuga menziesii, and Tsuga heterophylla), only Laccaria laccata and Cenococcum geophilum...

Non-anthocyanin polyphenolic transformation by native yeast and bacteria co-inoculation strategy during vinification.

PubMed

Devi, Apramita; Archana, Kodira Muthanna; Bhavya, Panikuttria Kuttappa; Anu-Appaiah, Konerira Aiyappaa

2018-02-01

Co-inoculation has been adapted by many wine-producing countries because it enhances the success of malolactic fermentation and reduces the fermentation cost, as well as time. However, wine phenolics have been sparsely highlighted during co-inoculation, even though polyphenols are an important parameter affecting wine colour, astringency and aroma. In the present study, we investigated the impact of co-inoculation on non-anthocyanin polyphenol profile for two different grape varieties. Co-inoculation of native yeast strain (AAV2) along with Oenococcus oeni was adapted for Cabernet Sauvignon and Shiraz wine. It was observed that the co-inoculation had minimal yet significant impact on the phenolic composition of wines for both the grape varieties. Color loss, as well as fruity aroma development, was observed in co-inoculated wines. The wines were on a par with the commercial wine, as well as wines without malolactic fermentation, in terms of phenolic compounds and overall organoleptic acceptance. Principal component analysis and hierarchical cluster analysis further suggested that the varietal influence on phenolic composition was dominating compared to inoculation strategies. Among the varieties, the inoculation strategies have significantly influenced the Cabernet wines compared to Shiraz wines. The results of the present study demonstrate that the phenolic compounds are not drastically affected by metabolic activities of malolactic bacteria during co-inoculation and, hence, are equally suitable for wine fermentation. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Enhancement of clover growth by inoculation of P-solubilizing fungi and arbuscular mycorrhizal fungi.

PubMed

Souchie, Edson L; Azcón, Rosario; Barea, Jose M; Silva, Eliane M R; Saggin-Júnior, Orivaldo J

2010-09-01

This study evaluated the synergism between several P-solubilizing fungi isolates and arbuscular mycorrhizal fungi to improve clover ( Trifolium pratense) growth in the presence of Araxá apatite. Clover was sown directly in plastic pots with 300g of sterilized washed sand, vermiculite and sepiolite 1:1:1 (v:v:v) as substrate, and grown in a controlled environment chamber. The substrate was fertilized with 3 g L(-1) of Araxá apatite. A completely randomized design, in 8×2 factorial scheme (eight P-solubilizing fungi treatments with or without arbuscular mycorrhizal fungi)and four replicates were used. The P-solubilizing fungi treatments consisted of five Brazilian P-solubilizing fungi isolates (PSF 7, 9, 20, 21 and 22), two Spanish isolates ( Aspergillus niger and the yeast Yarowia lipolytica) and control (non-inoculated treatment). The greatest clover growth rate was recorded when Aspergillus niger and PSF 21 were co-inoculated with arbuscular mycorrhizal fungi. Aspergillus niger, PSF 7 and PSF 21 were the most effective isolates on increasing clover growth in the presence of arbuscular mycorrhizal fungi. Greater mycorrhizal colonization resulted in greater clover growth rate in most PSF treatments. PSF 7 was the best isolate to improve the establishment of mycorrhizal and rhizobia symbiosis.

Rabies neutralizing antibody response to different schedules of serum and vaccine inoculations in non-exposed persons

PubMed Central

Atanasiu, P.; Bahmanyar, M.; Baltazard, M.; Fox, J. P.; Habel, K.; Kaplan, M. M.; Kissling, R. E.; Komarov, A.; Koprowski, H.; Lépine, P.; Gallardo, F. Pérez; Schaeffer, M.

1957-01-01

Further studies were made on groups of adult humans, previously unexposed to rabies and with no history of rabies vaccination, who were inoculated with different schedules of phenolized inactivated vaccine given subcutaneously and high egg passage (HEP) Flury strain vaccine given intradermally, with and without inoculation of antirabies serum. Serum specimens of the inoculated individuals were studied for antibody up to the 60th day after the first inoculation of the vaccines and serum. Studies were also made on the effect of “booster” doses of HEP Flury strain vaccine given 6 months after preparatory inoculations. The results can be summarized as follows: 1. Fourteen daily inoculations of phenolized vaccine produced a superior antibody response to that obtained with 3 inoculations given 5 days apart. 2. Three intradermal inoculations of HEP Flury vaccine given 5 days apart gave a low level of antibody response, but these individuals responded efficiently by producing antibody to a “booster” dose of the same vaccine given 6 months later. 3. Administration of phenolized vaccine or of HEP Flury vaccine alone did not produce detectable antibody in most individuals until between the 10th and the 15th day after the first inoculation of the vaccine. 4. Passive antibody following inoculation of antirabies serum persisted in some individuals for as long as 42 days. Two inoculations of serum administered 5 days apart did not give levels of antibody higher than those obtained with one inoculation. 5. One inoculation of serum completely suppressed antibody response to 3 inoculations of Flury vaccine given intradermally 5 days apart, and also prevented the preparation of the individuals to respond to a later “booster” dose of this vaccine. 6. Three inoculation of phenolized vaccine given 5 days apart acted efficiently in producing antibody by the 60th day. However, the interfering action of one and two inoculations of serum was clearly defined in this schedule. 7

A new strategy for co-composting dairy manure with rice straw: Addition of different inocula at three stages of composting.

PubMed

Zhou, Cheng; Liu, Zhang; Huang, Zhao-Lin; Dong, Ming; Yu, Xiao-Long; Ning, Ping

2015-06-01

In considering the impact of inoculation time and the characteristics of composting material and inoculants on the usefulness of inoculation, a new composting strategy has been proposed and studied, in which three inocula were inoculated at three stages of composting process respectively: inoculum A (Thermoactinomyces sp. GF1 and GF2) was inoculated before fermentation to increase or maintain high temperature of pile, inoculum B (Coprinus cinerea and Coprinus comatus) was inoculated after thermophilic phase to promote degradation of lignin, and inoculum C (Trichoderma harzianum and Rhizopus oryzae) was inoculated after 30-day fermentation to promote degradation of cellulose. The results showed that the inoculations could significantly enhance the temperature of pile and the degradation of lignocelluloses. When inocula A, B, and C were inoculated into pile, temperature increased from 25°C to 65°C, from 33°C to 39°C and from 33°C to 38°C respectively and 35% lignin and 43% cellulose had been degraded in inoculated pile compared to the degradation of 15% lignin and 25% cellulose in control pile. As a result, the C/N ratio dropped more rapidly degraded in the inoculated pile (reached 20 after 33-day fermentation) than that in the control pile (reached 21.7 after 45-day fermentation). In addition, the volume loss in inoculated pile (76.5%) was higher than that in control pile (53.2%). The study, therefore, indicated that inoculating proper microorganisms at appropriate time improved the composting process and our new composting strategy would be propitious to the co-composting dairy manure with rice straw. Copyright © 2015 Elsevier Ltd. All rights reserved.

Response of Sesbania grandiflora to Inoculation of Soil with Vesicular-Arbuscular Mycorrhizal Fungi.

PubMed

Habte, M; Aziz, T

1985-09-01

A greenhouse experiment was conducted to determine the influence of two tropical isolates of Glomus fasciculatum and Glomus mosseae on the nutrient uptake and growth of Sesbania grandiflora. Inoculation of sterile soil with the fungi significantly improved growth and nutrient uptake by S. grandiflora, but the response of the legume was markedly better when the soil was inoculated with G. fasciculatum than when it was inoculated with G. mosseae. Nutrient uptake and growth of S. grandiflora in nonsterile soil was also significantly stimulated by inoculation, but the legume did not respond differently to the two endophytes under this condition.

Milk production response to feeding alfalfa silage inoculated with Lactobacillus plantarum

USDA-ARS?s Scientific Manuscript database

In mini-silo trials, silages treated with a Lactobacillus plantarum silage inoculant (Ecosyl, Yorkshire, UK) had increased in vitro rumen microbial biomass production compared to untreated. Our objective was to determine if alfalfa silage treated with this inoculant could produce a milk production r...

IL-22 from T cells: better late than never.

PubMed

Honda, Kenya

2012-12-14

Interleukin-22 (IL-22) enhances mucosal barrier function and is important in antimicrobial host defense. In this issue of Immunity, Basu et al. (2012) reveal that Th22 cells are the critical adaptive source of IL-22 during late-phase infection by Citrobacter rodentium. Copyright © 2012 Elsevier Inc. All rights reserved.

Azolla filiculoides Nitrogenase Activity Decrease Induced by Inoculation with Chlamydomonas sp. †

PubMed Central

Habte, Mitiku

1986-01-01

Experiments were conducted to determine the influence of Chlamydomonas sp. on nitrogen fixation (C2H2 → C2H4) in Azolla filiculoides and on the nitrogen fixation and growth of free-living Anabaena azollae 2B organisms. Inoculation of azolla medium with Chlamydomonas sp. was associated with decreased nitrogenase activity in A. filiculoides and with increases in the density of a fungal population identified as Acremonium sp. Subsequent inoculation of azolla medium with this fungus was also accompanied by a significant decrease in nitrogenase activity of A. filiculoides. However, the extent of depression of nitrogenase activity was significantly higher when azolla medium was inoculated with Chlamydomonas sp. than when it was inoculated with Acremonium sp. Inoculation of nitrogen-free Stanier medium with either Acremonium sp. or Chlamydomonas sp. did not adversely affect the growth or nitrogenase activity of free-living A. azollae. Decreased nitrogenase activity in A. filiculoides is apparently related to the adverse influence of the green alga and the fungus on the macrosymbiont. The mechanisms that might be involved are discussed. PMID:16347211

Inoculation with arbuscular mycorrhizal fungi improves the nutritional value of tomatoes.

PubMed

Hart, Miranda; Ehret, David L; Krumbein, Angelika; Leung, Connie; Murch, Susan; Turi, Christina; Franken, Philipp

2015-07-01

Arbuscular mycorrhizal (AM) fungi can affect many different micronutrients and macronutrients in plants and also influence host volatile compound synthesis. Their effect on the edible portions of plants is less clear. Two separate studies were performed to investigate whether inoculation by AM fungi (Rhizophagus irregularis, Funneliformis mosseae, or both) can affect the food quality of tomato fruits, in particular common minerals, antioxidants, carotenoids, a suite of vitamins, and flavor compounds (sugars, titratable acids, volatile compounds). It was found that AM fungal inoculation increased the nutrient quality of tomato fruits for most nutrients except vitamins. Fruit mineral concentration increased with inoculation (particularly N, P, and Cu). Similarly, inoculated plants had fruit with higher antioxidant capacity and more carotenoids. Furthermore, five volatile compounds were significantly higher in AM plants compared with non-AM controls. Taken together, these results show that AM fungi represent a promising resource for improving both sustainable food production and human nutritional needs.

The roles of inoculants' carbon source use in the biocontrol of potato scab disease.

PubMed

Sun, Pingping; Zhao, Xinbei; Shangguan, Nini; Chang, Dongwei; Ma, Qing

2015-04-01

Despite the application of multiple strains in the biocontrol of plant diseases, multistrain inoculation is still constrained by its inconsistency in the field. Nutrients, especially carbons, play an important role in the biocontrol processes. However, little work has been done on the systematic estimation of inoculants' carbon source use on biocontrol efficacies in vivo. In the present study, 7 nonpathogenic Streptomyces strains alone and in different combinations were inoculated as biocontrol agents against the potato scab disease, under field conditions and greenhouse treatments. The influence of the inoculants' carbon source use properties on biocontrol efficacies was investigated. The results showed that increasing the number of inoculated strains did not necessarily result in greater biocontrol efficacy in vivo. However, single strains with higher growth rates or multiple strains with less carbon source competition had positive effects on the biocontrol efficacies. These findings may shed light on optimizing the consistent biocontrol of plant disease with the consideration of inoculants' carbon source use properties.

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.

PubMed

Malmierca, M G; McCormick, S P; Cardoza, R E; Monte, E; Alexander, N J; Gutiérrez, S

2015-11-01

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.

Antipathy of Trichoderma against Sclerotium rolfsii Sacc.: Evaluation of Cell Wall-Degrading Enzymatic Activities and Molecular Diversity Analysis of Antagonists.

PubMed

Hirpara, Darshna G; Gajera, Harsukh P; Hirpara, Hitesh Z; Golakiya, Balubhai A

2017-01-01

The fungus Trichoderma is a teleomorph of the Hypocrea genus and associated with biological control of plant diseases. The microscopic, biochemical, and molecular characterization of Trichoderma was carried out and evaluated for in vitro antagonistic activity against the fungal pathogen Sclerotium rolfsii causing stem rot disease in groundnut. In total, 11 isolates of Trichoderma were examined for antagonism at 6 and 12 days after inoculation (DAI). Out of 11, T. virens NBAII Tvs12 evidenced the highest (87.91%) growth inhibition of the test pathogen followed by T. koningii MTCC 796 (67.03%), T. viride NBAII Tv23 (63.74%), and T. harzianum NBAII Th1 (60.44%). Strong mycoparasitism was observed in the best antagonist Tvs12 strain during 6-12 DAI. The specific activity of cell wall-degrading enzymes - chitinase and β-1,3-glucanase - was positively correlated with growth inhibition of the test pathogen. In total, 18 simple sequence repeat (SSR) polymorphisms were reported to amplify 202 alleles across 11 Trichoderma isolates. The average polymorphism information content for SSR markers was found to be 0.80. The best antagonist Tvs 12 was identified with 7 unique SSR alleles amplified by 5 SSR markers. Clustering patterns of 11 Trichoderma strains showed the best antagonist T. virens NBAII Tvs 12 outgrouped with a minimum 3% similarity from the rest of Trichoderma. © 2017 S. Karger AG, Basel.

Synthesis and X-ray structures of dilithium complexes of the phosphonate anions [PhP(E)(N(t)Bu)(2)](2-) (E = O, S, Se, Te) and dimethylaluminum derivatives of [PhP(E)(N(t)Bu)(NH(t)Bu)](-) (E = S, Se).

PubMed

Briand, Glen G; Chivers, Tristram; Krahn, Mark; Parvez, Masood

2002-12-16

The dilithium salts of the phosphonate dianions [PhP(E)(N(t)Bu)(2)](2-) (E = O, S, Se) are generated by the lithiation of [PhP(E)(NH(t)Bu)(2)] with n-butyllithium. The formation of the corresponding telluride (E = Te) is achieved by oxidation of [Li(2)[PhP(N(t)Bu)(2)

Acetylene reduction (nitrogen fixation) associated with corn inoculated with Spirillum.

PubMed

Barber, L E; Tjepkema, J D; Russell, S A; Evans, H J

1976-07-01

Sorghum and corn breeding lines were grown in soil in field and greenhouse experiments with and without an inoculum of N2-fixing in Spirillum strains from Brazil. Estimated rates of N2 fixation associated with field-grown corn and sorghum plants were less than 4 g of N2/ha per day. The mean estimated N2-fixation rates determined on segments of roots from corn inoculated with Spirillum and grown in the greenhouse at 24 to 27 degrees C were 15 g of N2/ha per day (16 inbreds), 25 g of N2/ha per day (six hybrids), and 165 g of N2/ha per day for one hybird which was heavily inoculated. The corresponding mean rates determined from measurements of in situ cultures of the same series of corn plants (i.e., 16 inbreds, six hybrids, and one heavily inoculated hybrid) were 0.4, 2.3, and 1.1 g of N2/ha per day, respectively. Lower rates of C2H2 reduction were associated with control corn cultures which had been treated with autoclaved Spirillum than with cultures inoculated with live Spirillum. No C2H2 reduction was detected in plant cultures treated with ammonium nitrate. Numbers of nitrogen-fixing bacteria on excised roots of corn plants increased an average of about 30-fold during an overnight preincubation period, and as a result acetylene reduction assays of root samples after preincubation failed to serve as a valid basis for estimating N2 fixation by corn in pot cultures. Plants grown without added nitrogen either with or without inoculum exhibited severe symptoms of nitrogen deficiency and in most cases produced significantly less dry weight than those supplied with fixed nitrogen. Although substantial rates of C2H2 reduction by excised corn roots were observed after preincubation under limited oxygen, the yield and nitrogen content of inoculated plants and the C2H2-reduction rates by inoculated pot cultures of corn, in situ, provided no evidence of appreciable N2 fixation.

Use of Torulaspora delbrueckii and Saccharomyces cerevisiae in semi-industrial sequential inoculation to improve quality of Palomino and Chardonnay wines in warm climates.

PubMed

Puertas, B; Jiménez, M J; Cantos-Villar, E; Cantoral, J M; Rodríguez, M E

2017-03-01

We have evaluated for the first time the impact of two commercial yeast strains (Torulaspora delbrueckii TD291 and Saccharomyces cerevisiae QA23) inoculated sequentially in musts of Chardonnay and Palomino Fino grape varieties grown under warm climate (South-west of Spain). Semi-industrial scale alcoholic fermentations (AF) were performed during the 2011 and 2012 harvests. Implantation analyses demonstrated that T. delbrueckii is the predominant strain until the end of the AF phase. Wines with sequential inoculation (SI) resulted in the production of low levels of acetic acid (which gives wine an undesirable 'vinegary' character), low acetaldehyde in Chardonnay and high in Palomino wines. The most salient attributes that contribute to the quality of the Chardonnay and Palomino wines produced were aroma intensity, fresh and tropical fruit character. This study demonstrated that SI of T. delbrueckii and S. cerevisiae contribute significantly to the improvement of Chardonnay wine aromas and the creation of new styles of wine for Palomino. This study has generated new knowledge about the biotechnological potential of T. delbrueckii (TD219) and S. cerevisiae (QA23) for improving the organoleptic properties of Chardonnay and Palomino wines. © 2016 The Society for Applied Microbiology.

Structural basis of transport function in major facilitator superfamily protein from Trichoderma harzianum.

PubMed

Chaudhary, Nitika; Sandhu, Padmani; Ahmed, Mushtaq; Akhter, Yusuf

2017-02-01

Trichothecenes are the sesquiterpenes secreted by Trichoderma spp. residing in the rhizosphere. These compounds have been reported to act as plant growth promoters and bio-control agents. The structural knowledge for the transporter proteins of their efflux remained limited. In this study, three-dimensional structure of Thmfs1 protein, a trichothecene transporter from Trichoderma harzianum, was homology modelled and further Molecular Dynamics (MD) simulations were used to decipher its mechanism. Fourteen transmembrane helices of Thmfs1 protein are observed contributing to an inward-open conformation. The transport channel and ligand binding sites in Thmfs1 are identified based on heuristic, iterative algorithm and structural alignment with homologous proteins. MD simulations were performed to reveal the differential structural behaviour occurring in the ligand free and ligand bound forms. We found that two discrete trichothecene binding sites are located on either side of the central transport tunnel running from the cytoplasmic side to the extracellular side across the Thmfs1 protein. Detailed analysis of the MD trajectories showed an alternative access mechanism between N and C-terminal domains contributing to its function. These results also demonstrate that the transport of trichodermin occurs via hopping mechanism in which the substrate molecule jumps from one binding site to another lining the transport tunnel. Copyright © 2016 Elsevier B.V. All rights reserved.

Trichoderma harzianum Produces a New Thermally Stable Acid Phosphatase, with Potential for Biotechnological Application

PubMed Central

Souza, Amanda Araújo; Leitão, Vanessa Oliveira; Ramada, Marcelo Henrique; Mehdad, Azadeh; Georg, Raphaela de Castro; Ulhôa, Cirano José; de Freitas, Sonia Maria

2016-01-01

Acid phosphatases (ACPases) are produced by a variety of fungi and have gained attention due their biotechnological potential in industrial, diagnosis and bioremediation processes. These enzymes play a specific role in scavenging, mobilization and acquisition of phosphate, enhancing soil fertility and plant growth. In this study, a new ACPase from Trichoderma harzianum, named ACPase II, was purified and characterized as a glycoprotein belonging to the acid phosphatase family. ACPase II presents an optimum pH and temperature of 3.8 and 65°C, respectively, and is stable at 55°C for 120 min, retaining 60% of its activity. The enzyme did not require metal divalent ions, but was inhibited by inorganic phosphate and tungstate. Affinity for several phosphate substrates was observed, including phytate, which is the major component of phosphorus in plant foods. The inhibition of ACPase II by tungstate and phosphate at different pH values is consistent with the inability of the substrate to occupy its active site due to electrostatic contacts that promote conformational changes, as indicated by fluorescence spectroscopy. A higher affinity for tungstate rather than phosphate at pH 4.0was observed, in accordance with its highest inhibitory effect. Results indicate considerable biotechnological potential of the ACPase II in soil environments. PMID:26938873

Trichoderma harzianum Produces a New Thermally Stable Acid Phosphatase, with Potential for Biotechnological Application.

PubMed

Souza, Amanda Araújo; Leitão, Vanessa Oliveira; Ramada, Marcelo Henrique; Mehdad, Azadeh; Georg, Raphaela de Castro; Ulhôa, Cirano José; de Freitas, Sonia Maria

2016-01-01

Acid phosphatases (ACPases) are produced by a variety of fungi and have gained attention due their biotechnological potential in industrial, diagnosis and bioremediation processes. These enzymes play a specific role in scavenging, mobilization and acquisition of phosphate, enhancing soil fertility and plant growth. In this study, a new ACPase from Trichoderma harzianum, named ACPase II, was purified and characterized as a glycoprotein belonging to the acid phosphatase family. ACPase II presents an optimum pH and temperature of 3.8 and 65 °C, respectively, and is stable at 55 °C for 120 min, retaining 60% of its activity. The enzyme did not require metal divalent ions, but was inhibited by inorganic phosphate and tungstate. Affinity for several phosphate substrates was observed, including phytate, which is the major component of phosphorus in plant foods. The inhibition of ACPase II by tungstate and phosphate at different pH values is consistent with the inability of the substrate to occupy its active site due to electrostatic contacts that promote conformational changes, as indicated by fluorescence spectroscopy. A higher affinity for tungstate rather than phosphate at pH 4.0 was observed, in accordance with its highest inhibitory effect. Results indicate considerable biotechnological potential of the ACPase II in soil environments.

A patient presenting a 22q13 deletion associated with an apparently balanced translocation t(16;22): An illustrative case in the investigation of patients with low ARSA activity

PubMed Central

Artigalás, Osvaldo; Paskulin, Giorgio; Riegel, Mariluce; Burin, Maira; Saraiva-Pereira, Maria Luiza; Maluf, Sharbel; Kiss, Andrea; Schwartz, Ida Vanessa D.

2012-01-01

A 10-year-old speechless, mentally deficient male, with low arylsulfatase A (ARSA) activity, and presumably, methachromatic leukodystrophy, underwent genetic evaluation. As the clinical picture was not compatible with this diagnosisan ARSA gene and chromosome analysis were performed, showing the presence of a pseudodeficiency ARSA allele and a de novo apparently balanced t(16;22)(p11.2;q13) translocation. A deletion on the long arm of chromosome 22 encompassing the ARSA gene, as shown by FISH and array-CGH, indicated a 22q13 deletion syndrome. This case illustrates the importance of detailed cytogenetic investigation in patients presenting low arylsulfatase A activity and atypical/unspecific clinical features. PMID:22888290

Expressed sequence tags related to nitrogen metabolism in maize inoculated with Azospirillum brasilense.

PubMed

Pereira-Defilippi, L; Pereira, E M; Silva, F M; Moro, G V

2017-05-31

The relative quantitative real-time expression of two expressed sequence tags (ESTs) codifying for key enzymes in nitrogen metabolism in maize, nitrate reductase (ZmNR), and glutamine synthetase (ZmGln1-3) was performed for genotypes inoculated with Azospirillum brasilense. Two commercial single-cross hybrids (AG7098 and 2B707) and two experimental synthetic varieties (V2 and V4) were raised under controlled greenhouse conditions, in six treatment groups corresponding to different forms of inoculation and different levels of nitrogen application by top-dressing. The genotypes presented distinct responses to inoculation with A. brasilense. Increases in the expression of ZmNR were observed for the hybrids, while V4 only displayed a greater level of expression when the plants received nitrogenous fertilization by top-dressing and there was no inoculation. The expression of the ZmGln1-3EST was induced by A. brasilense in the hybrids and the variety V4. In contrast, the variety V2 did not respond to inoculation.

Inoculation of Scytalidium thermophilum in Button Mushroom Compost and Its Effect on Yield.

PubMed

Straatsma, G; Olijnsma, T W; Gerrits, J P; Amsing, J G; Op Den Camp, H J; Van Griensven, L J

1994-09-01

Scytalidium thermophilum isolates in culture, as well as the endogenous strain(s) in mushroom compost, were inactivated at 70 degrees C. This temperature was used to pasteurize composts for experiments. Of nine thermophilic fungal species, only S. thermophilum and Myriococcum thermophilum grew well on pasteurized compost in test tubes. The effect of both species on the crop yield of Agaricus bisporus mushrooms was studied. In solid-state fermentation rooms called tunnels, compost was pasteurized and inoculated. After incubation, the inoculated organisms were reisolated and counted, showing their successful colonization. The yield of mushrooms on inoculated composts was almost twice that on the pasteurized control. This result demonstrates the effectiveness of S. thermophilum in compost preparation. Inoculation is not necessary for traditional compost preparation. Naturally occurring strains of S. thermophilum, present in ingredients, readily colonize compost during preparation. Inoculation may be vital if compost is pretreated at a high temperature in tunnels. This finding is of relevance for the environmentally controlled production of high-yielding compost.

Inoculation of Scytalidium thermophilum in Button Mushroom Compost and Its Effect on Yield

PubMed Central

Straatsma, Gerben; Olijnsma, Tineke W.; Gerrits, Jan P. G.; Amsing, Jos G. M.; Op Den Camp, Huub J. M.; Van Griensven, Leo J. L. D.

1994-01-01

Scytalidium thermophilum isolates in culture, as well as the endogenous strain(s) in mushroom compost, were inactivated at 70°C. This temperature was used to pasteurize composts for experiments. Of nine thermophilic fungal species, only S. thermophilum and Myriococcum thermophilum grew well on pasteurized compost in test tubes. The effect of both species on the crop yield of Agaricus bisporus mushrooms was studied. In solid-state fermentation rooms called tunnels, compost was pasteurized and inoculated. After incubation, the inoculated organisms were reisolated and counted, showing their successful colonization. The yield of mushrooms on inoculated composts was almost twice that on the pasteurized control. This result demonstrates the effectiveness of S. thermophilum in compost preparation. Inoculation is not necessary for traditional compost preparation. Naturally occurring strains of S. thermophilum, present in ingredients, readily colonize compost during preparation. Inoculation may be vital if compost is pretreated at a high temperature in tunnels. This finding is of relevance for the environmentally controlled production of high-yielding compost. PMID:16349366

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system

PubMed Central

Rossi-Rodrigues, Bianca Caroline; Brochetto-Braga, Márcia Regina; Tauk-Tornisielo, Sâmia Maria; Carmona, Eleonora Cano; Arruda, Valeska Marques; Chaud Netto, José

2009-01-01

Trichoderma is one of the fungi genera that produce important metabolites for industry. The growth of these organisms is a consequence of the nutritional sources used as also of the physical conditions employed to cultivate them. In this work, the automated Bioscreen C system was used to evaluate the influence of different nutritional sources on the growth of Trichoderma strains (T. hamatum, T. harzianum, T. viride, and T. longibrachiatum) isolated from the soil in the Juréia-Itatins Ecological Station (JIES), São Paulo State - Brazil. The cultures were grown in liquid culture media containing different carbon- (2%; w/v) and nitrogen (1%; w/v) sources at 28ºC, pH 6.5, and agitated at 150 rpm for 72 h. The results showed, as expected, that glucose is superior to sucrose as a growth-stimulating carbon source in the Trichoderma strains studied, while yeast extract and tryptone were good growth-stimulating nitrogen sources in the cultivation of T. hamatum and T. harzianum. PMID:24031380

Effects of inoculation timing on symptom development in Ulmus americana L.

Treesearch

Garrett L. Beier; Benjamin W. Held; Chad P. Giblin; Robert A. Blanchette

2017-01-01

Field inoculation trials are an important component of screening American elms (Ulmus americana) for levels of resistance to Dutch elm disease. A major concern in screening is variability in disease ratings from year to year. Previous studies have demonstrated that timing of inoculation can have a significant impact on disease susceptibility. In...

Effect of thermo-tolerant actinomycetes inoculation on cellulose degradation and the formation of humic substances during composting.

PubMed

Zhao, Yi; Zhao, Yue; Zhang, Zhechao; Wei, Yuquan; Wang, Huan; Lu, Qian; Li, Yanjie; Wei, Zimin

2017-10-01

The inoculum containing four cellulolytic thermophilic actinomycetes was screened from compost samples, and was inoculated into co-composting during different inoculation phases. The effect of different inoculation phases on cellulose degradation, humic substances formation and the relationship between inoculation and physical-chemical parameters was determined. The results revealed that inoculation at different phases of composting improved cellulase activities, accelerated the degradation of cellulose, increased the content of humic substances and influenced the structure of actinomycetic community, but there were significant differences between different inoculation phases. Redundancy analysis showed that the different inoculation phases had different impacts on the relationship between exogenous actinobacteria and physical-chemical parameters. Therefore, based on the promoting effort of inoculation in thermophilic phase of composting for the formation of humic substances, we suggested an optimized inoculation strategy to increase the content of humic substances, alleviate CO 2 emission during composting. Copyright © 2017 Elsevier Ltd. All rights reserved.

The Impact of Inoculation Parameters on the Pathogenesis of Contact Lens–Related Infectious Keratitis

PubMed Central

Tam, Connie; Mun, James J.; Evans, David J.

2010-01-01

Purpose. Contact lens wear predisposes to Pseudomonas aeruginosa keratitis, but the mechanisms involved remain unclear. An in vivo model was used to study lens inoculation conditions enabling disease. Methods. Custom-made hydrogel contact lenses were fitted to rats after incubation in P. aeruginosa approximately 1011 cfu/mL (3 hours) or approximately 103 cfu/mL (24 hours). Another group was inadvertently inoculated with a suction pen previously used with high inocula, but rinsed in ethanol and stored dry (6 months). Some corneas were tissue paper–blotted to cause fluorescein staining before lens fitting. Contralateral eyes were untreated. Twenty-four hours after disease detection, lenses were transferred to naive rats or examined by confocal microscopy before homogenization to quantify viable bacteria. After lens removal, corneas were washed to collect nonadherent bacteria and were analyzed by immunohistochemistry. Results. All eyes challenged with unworn contaminated lenses developed keratitis after approximately 7 to 10 days. Disease delay and severity were unaffected by inoculum parameters or tissue blotting but occurred sooner with lenses transferred from infected eyes (∼2 days). Worn lenses and corneal washes contained infecting bacteria. Posterior, not anterior, lens surfaces harbored P. aeruginosa biofilms that penetrated the lens matrix. Diseased corneas showed an infiltration of phagocytes and T-lymphocytes. Conclusions. P. aeruginosa induces keratitis in this lens-wearing model after a single inoculation. Delayed disease onset was interesting considering the greater keratitis risk during extended wear. Infection did not require the disruption of corneal barrier function before lens wear and occurred without exposure to lens care solutions. The data suggest that keratitis involves biofilm formation or other bacterial adaptations in vivo. PMID:20130275

Inoculation testing of Apollo 12 materials

NASA Technical Reports Server (NTRS)

1969-01-01

E. Landrum Young, Brown and Root Northrop, injects a young Japanese quail with a suspension of pulvarized Apollo 12 lunar material within a quarantine cabinet in the Invertebrate, Aves and Fish Laboratory of the Lunar Receiving Laboratory, bldg 37, Manned Spacecraft Center. The bird is being inoculated in the abdominal cavity.

Rethinking the Inoculation Analogy: Effects on Subjects with Differing Preexisting Attitudes

ERIC Educational Resources Information Center

Wood, Michelle L. M.

2007-01-01

Inoculation messages employed in past studies have been consistently preventative. Yet, if inoculation strategies are to be used in mass media campaigns, researchers need to know what the effects will be on all audience members--not just those known to support a message sponsor's position. A 3-phase experiment was conducted involving 558…

C22-bronchial and T7-alveolar epithelial cell lines of the immortomouse are excellent murine cell culture model systems to study pulmonary peroxisome biology and metabolism.

PubMed

Karnati, Srikanth; Palaniswamy, Saranya; Alam, Mohammad Rashedul; Oruqaj, Gani; Stamme, Cordula; Baumgart-Vogt, Eveline

2016-03-01

In pulmonary research, temperature-sensitive immortalized cell lines derived from the lung of the "immortomouse" (H-2k(b)-tsA58 transgenic mouse), such as C22 club cells and T7 alveolar epithelial cells type II (AECII), are frequently used cell culture models to study CC10 metabolism and surfactant synthesis. Even though peroxisomes are highly abundant in club cells and AECII and might fulfill important metabolic functions therein, these organelles have never been investigated in C22 and T7 cells. Therefore, we have characterized the peroxisomal compartment and its associated gene transcription in these cell lines. Our results show that peroxisomes are highly abundant in C22 and T7 cells, harboring a common set of enzymes, however, exhibiting specific differences in protein composition and gene expression patterns, similar to the ones observed in club cells and AECII in situ in the lung. C22 cells contain a lower number of larger peroxisomes, whereas T7 cells possess more numerous tubular peroxisomes, reflected also by higher levels of PEX11 proteins. Moreover, C22 cells harbor relatively higher amounts of catalase and antioxidative enzymes in distinct subcellular compartments, whereas T7 cells exhibit higher levels of ABCD3 and plasmalogen synthesizing enzymes as well as nuclear receptors of the PPAR family. This study suggest that the C22 and T7 cell lines of the immortomouse lung are useful models to study the regulation and metabolic function of the peroxisomal compartment and its alterations by paracrine factors in club cells and AECII.

Survival and recovery of Listeria monocytogenes on ready-to-eat meats inoculated with a desiccated and nutritionally depleted dustlike vector.

PubMed

De Roin, Mark A; Foong, Sally C C; Dixon, Philip M; Dickson, James S

2003-06-01

Dust from construction was theorized to serve as a vector for L. monocytogenes transmission to ready-to-eat (RTE) meats after heat processing but before packaging. A five-strain Listeria monocytogenes culture including serotype 4b was continually stressed on a sand vector under four sets of nutritionally depleted and dry conditions to simulate postprocessing contamination by dustlike particulates. The stresses included that associated with sand stored at different temperatures (10 and 22 degrees C) and levels of humidity (40% relative humidity [RH], 88% RH, or complete desiccation). Irradiated RTE meats, including frankfurters, bologna, chopped ham, and deli-style roast beef, were inoculated with the L. monocytogenes-contaminated sand every 2 to 3 days over a period of 1 1/2 months. After inoculation, the RTE meats were vacuum packed and stored at 4 degrees C for 24 h. Populations of L. monocytogenes were enumerated by surface plating on nonselective and selective media to recover cells on the basis of the different stresses presented (osmotic or antibiotic). L. monocytogenes was demonstrated to be capable of surviving on the sand vector for > 151 days at 10 degrees C and 88% RH, 136 days at 10 degrees C and 0% RH, 73 days at 22 degrees C and 40% RH, and 82 days at 22 degrees C and 0% RH. These results show that under the most conservative scenario, the 73-day-old L. monocytogenes-contaminated sand was able to attach to and be recovered from the RTE meats. This study illustrated that dust contaminated with L. monocytogenes, once in contact with meat surfaces, can survive and grow, posing a health hazard to consumers.

Growth-dependent catalase localization in Exiguobacterium oxidotolerans T-2-2T reflected by catalase activity of cells.

PubMed

Hanaoka, Yoshiko; Takebe, Fumihiko; Nodasaka, Yoshinobu; Hara, Isao; Matsuyama, Hidetoshi; Yumoto, Isao

2013-01-01

A psychrotolerant and H2O2-resistant bacterium, Exiguobacterium oxidotolerans T-2-2(T), exhibits extraordinary H2O2 resistance and produces catalase not only intracellularly but also extracellularly. The intracellular and extracellular catalases exhibited the same enzymatic characteristics, that is, they exhibited the temperature-dependent activity characteristic of a cold-adapted enzyme, their heat stabilities were similar to those of mesophilic enzymes and very high catalytic intensity. In addition, catalase gene analysis indicated that the bacterium possessed the sole clade 1 catalase gene corresponding to intracellular catalase. Hence, intracellular catalase is secreted into the extracellular space. In addition to intracellular and extracellular catalases, the inner circumference of the cells showed the localization of catalase in the mid-stationary growth phase, which was observed by immunoelectron microscopy using an antibody against the intracellular catalase of the strain. The cells demonstrated higher catalase activity in the mid-stationary growth phase than in the exponential growth phase. The catalase localized in the inner circumference can be dissociated by treatment with Tween 60. Thus, the localized catalase is not tightly bound to the inner circumference of the cells and may play a role in the oxidative defense of the cells under low metabolic state.

Growth-Dependent Catalase Localization in Exiguobacterium oxidotolerans T-2-2T Reflected by Catalase Activity of Cells

PubMed Central

Hanaoka, Yoshiko; Takebe, Fumihiko; Nodasaka, Yoshinobu; Hara, Isao; Matsuyama, Hidetoshi; Yumoto, Isao

2013-01-01

A psychrotolerant and H2O2-resistant bacterium, Exiguobacterium oxidotolerans T-2-2T, exhibits extraordinary H2O2 resistance and produces catalase not only intracellularly but also extracellularly. The intracellular and extracellular catalases exhibited the same enzymatic characteristics, that is, they exhibited the temperature-dependent activity characteristic of a cold-adapted enzyme, their heat stabilities were similar to those of mesophilic enzymes and very high catalytic intensity. In addition, catalase gene analysis indicated that the bacterium possessed the sole clade 1 catalase gene corresponding to intracellular catalase. Hence, intracellular catalase is secreted into the extracellular space. In addition to intracellular and extracellular catalases, the inner circumference of the cells showed the localization of catalase in the mid-stationary growth phase, which was observed by immunoelectron microscopy using an antibody against the intracellular catalase of the strain. The cells demonstrated higher catalase activity in the mid-stationary growth phase than in the exponential growth phase. The catalase localized in the inner circumference can be dissociated by treatment with Tween 60. Thus, the localized catalase is not tightly bound to the inner circumference of the cells and may play a role in the oxidative defense of the cells under low metabolic state. PMID:24204687

Intake, digestibility, and composition of orchardgrass and alfalfa silages treated with cellulase, inoculant, and formic acid fed to lambs.

PubMed

Nadeau, E M; Russell, J R; Buxton, D R

2000-11-01

The objectives of this study were to determine the effect of a cellulase (from Trichoderma longibrachiatum) alone or combined with a bacterial inoculant (Lactobacillus plantarum and Pediococcus cerevisiae) or formic acid on composition, intake, and digestibility of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.) silages. Orchardgrass and alfalfa were harvested at the early heading stage and at the early bloom stage of maturity and wilted to approximately 22 and 32% DM, respectively. Forages were then ensiled in 100-L sealed barrels for at least 60 d before they were fed to lambs. Silage treated with cellulase had lower (P < .001) pH and lower (P < .001) acetic acid and NH3 N concentrations than untreated silage of both plant species and a higher (P = .004) lactic acid concentration than the control treatment of alfalfa silage. Fermentation characteristics of cellulase-treated silages, especially of alfalfa, were further enhanced by use of inoculant. Formic acid addition increased (P < .001), reducing sugar concentration of cellulase-treated orchardgrass and alfalfa silage by 90 and 154%, respectively, and decreased (P < .001) NH3 N concentration of cellulase-treated alfalfa silage by 19%. Averaged across plant species, cellulase, combined with inoculant or formic acid, resulted in 8 and 13% greater (P = .03) DMI, respectively, than the control silage. Extensive enzymatic cell-wall degradation during ensiling decreased (P = .003) NDF intake of cellulase-treated orchardgrass silage by 25% and decreased (P = .001) cellulose intake by 23%, when averaged across plant species. Addition of formic acid increased (P = .003) NDF intake of cellulase-treated orchardgrass silage by 19%. Averaged across species, cellulase application decreased (P < .05) silage NDF digestibility by 18%. Greater sugar and lower acetic acid, NH3 N, and NDF concentrations resulted in greater DMI of cellulase-treated silage than of control silage, when cellulase was combined

The Effect of Raffaelea quercus-mongolicae Inoculations on the Formation of Non-conductive Sapwood of Quercus mongolica.

PubMed

Torii, Masato; Matsuda, Yosuke; Seo, Sang Tae; Kim, Kyung Hee; Ito, Shin-Ichiro; Moon, Myung Jin; Kim, Seong Hwan; Yamada, Toshihiro

2014-06-01

In Korea, mass mortality of Quercus mongolica trees has become obvious since 2004. Raffaelea quercus-mongolicae is believed to be a causal fungus contributing the mortality. To evaluate the pathogenicity of the fungus to the trees, the fungus was multiple- and single-inoculated to the seedlings and twigs of the mature trees, respectively. In both the inoculations, the fungus was reisolated from more than 50% of inoculated twigs and seedlings. In the single inoculations, proportions of the transverse area of non-conductive sapwood at inoculation points and vertical lengths of discoloration expanded from the points were significantly different between the inoculation treatment and the control. In the multiple inoculations, no mortality was confirmed among the seedlings examined. These results showed that R. quercus-mongolicae can colonize sapwood, contribute to sapwood discoloration and disrupt sap flows around inoculation sites of Q. mongolica, although the pathogenicity of the fungus was not proven.

Stability of Bradyrhizobium japonicum Inoculants after Introduction into Soil

PubMed Central

Brunel, Brigitte; Cleyet-Marel, Jean-Claude; Normand, Philippe; Bardin, Rene

1988-01-01

Bradyrhizobium japonicum USDA 125-Sp, USDA 138, and USDA 138-Sm had been used as inoculants for soybean (Glycine max (L.) Merr.) in soils previously free of B. japonicum. At 8 to 13 years after their release, these strains were reisolated from soil samples. A total of 115 isolates were obtained through nodules, and seven colonies were obtained directly by a serological method. The stability of the inoculants was confirmed by comparing the reisolated cultures with their respective parental strains which had been preserved by being lyophilized or stored on a yeast extract-mannitol agar slant at 4°C. Comparisons were made on morphological and serological characters, carbon compound utilization (8 tested), intrinsic antibiotic resistance (9 tested), and enzymatic activity (19 tested). Mucous and nonmucous isolates of serogroup 125 were analyzed for symbiotic effectiveness and restriction fragment hybridization with a DNA probe. Our data suggest that the B. japonicum inoculants have survived for up to 13 years in the soils without significant mutation except for two reisolates with a slightly increased kanamycin resistance level. Images PMID:16347768

Beauveria bassiana and Metarhizium anisopliae endophytically colonize cassava roots following soil drench inoculation

PubMed Central

Greenfield, Melinda; Gómez-Jiménez, María I.; Ortiz, Viviana; Vega, Fernando E.; Kramer, Matthew; Parsa, Soroush

2016-01-01

We investigated the fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae to determine if endophytic colonization could be achieved in cassava. An inoculation method based on drenching the soil around cassava stem cuttings using conidial suspensions resulted in endophytic colonization of cassava roots by both entomopathogens, though neither was found in the leaves or stems of the treated cassava plants. Both fungal entomopathogens were detected more often in the proximal end of the root than in the distal end. Colonization levels of B. bassiana were higher when plants were sampled at 7–9 days post-inoculation (84%) compared to 47–49 days post-inoculation (40%). In contrast, the colonization levels of M. anisopliae remained constant from 7–9 days post-inoculation (80%) to 47–49 days post-inoculation (80%), which suggests M. anisopliae is better able to persist in the soil, or as an endophyte in cassava roots over time. Differences in colonization success and plant growth were found among the fungal entomopathogen treatments. PMID:27103778

The timing of TGF-β inhibition affects the generation of antigen-specific CD8+ T Cells

PubMed Central

2013-01-01

Background Transforming growth factor (TGF)-β is a potent immunosuppressive cytokine necessary for cancer growth. Animal and human studies have shown that pharmacologic inhibition of TGF-β slows the growth rate of established tumors and occasionally eradicates them altogether. We observed, paradoxically, that inhibiting TGF-β before exposing animals to tumor cells increases tumor growth kinetics. We hypothesized that TGF-β is necessary for the anti-tumor effects of cytotoxic CD8+ T lymphocytes (CTLs) during the early stages of tumor initiation. Methods BALB/c mice were pretreated with a blocking soluble TGF-β receptor (sTGF-βR, TGF-β-blockade group, n=20) or IgG2a (Control group, n=20) before tumor inoculation. Tumor size was followed for 6 weeks. In vivo lymphocyte assays and depletion experiments were then performed to investigate the immunological basis of our results. Lastly, animals were pretreated with either sTGF-βR (n=6) or IgG2a (n=6) prior to immunization with an adenoviral vector encoding the human papillomavirus E7 gene (Ad.E7). One week later, flow cytometry was utilized to measure the number of splenic E7-specific CD8+ T cells. Results Inhibition of TGF-β before the injection of tumor cells resulted in significantly larger average tumor volumes on days 11, 17, 22, 26 and 32 post tumor-inoculation (p < 0.05). This effect was due to the inhibition of CTLs, as it was not present in mice with severe combined immunodeficiency (SCID) or those depleted of CD8+ T cells. Furthermore, pretreatment with sTGF-βR inhibited tumor-specific CTL activity in a Winn Assay. Tumors grew to a much larger size when mixed with CD8+ T cells from mice pretreated with sTGF-βR than when mixed with CD8+ T cells from mice in the control group: 96 mm3 vs. 22.5 mm3, respectively (p < 0.05). In addition, fewer CD8+ T cells were generated in Ad.E7-immunized mice pretreated with sTGF-βR than in mice from the control group: 0.6% total CD8+ T cells vs. 1

Development and hydrology of biological soil crusts -- first results from a surface inoculation experiment

NASA Astrophysics Data System (ADS)

Mykhailova, Larysa; Raab, Thomas; Gypser, Stella; Fischer, Thomas

2016-04-01

Representing a set of various micro-biocoenoses, biocrusts often reside in adjacent patches, which not necessarily relate to structural elements of the habitat, like (micro-) topography or vegetational patterns. Such biocrust patches may become more stable through the formation of mutually dependent ecohydrological regimes. For example, algal patches inhibiting infiltration and generating runoff alternate with runoff-receiving moss patches possessing high water holding capacities. Here, we preliminarily report on a lysimeter field experiment where natural biocrust isolates were used for surface inoculation to (I) prove stochastic vs. deterministic biocrust development and (II) to quantitatively relate biocrust development to soil hydrology. Lysimeter sand was collected from 3-4 m below surface at natural dune outcrops in south-eastern Brandenburg, Germany (Glashütte (GLA) and Neuer Lugteich (LUG)), where biocrust samples were collected at the respective dune bases. The lysimeters were designed to prevent runoff. In a completely randomized full-factorial design, three factors were considered. (A) Inocolum in three treatments (bare control, mosses, algae), (B) mineral substrate texture in two treatments (GLA: 55% and LUG: 79% particles >630 μm), and (C) surface compaction in two treatments (control, 41.5 kN m-2 for 30 seconds). The samples were kept dry and re-moistened to -60 hPa two days before inoculation. After a species inventory, the inoculate was isolated by gently washing off sand particles from the biocrust samples. Algal/lichen crusts were dominated by Zygogonium ericetorum and Cladonia sp. at both sites. All moss crusts were dominated by Polytrichum piliferum and Ceratodon purpureus, whereas Brachythecium albicans was present at GLA only. 20 g of homogenized moist inoculate were spread over the surface of each lysimeter (Ø 19 cm, 22 cm depth). We performed autochthonous inoculation, i.e. biocrust isolates collected from GLA were used for inoculation of

Respiratory Francisella tularensis live vaccine strain infection induces Th17 cells and prostaglandin E2, which inhibits generation of gamma interferon-positive T cells.

PubMed

Woolard, Matthew D; Hensley, Lucinda L; Kawula, Thomas H; Frelinger, Jeffrey A

2008-06-01

Two key routes of Francisella tularensis infection are through the skin and airway. We wished to understand how the route of inoculation influenced the primary acute adaptive immune response. We show that an intranasal inoculation of the F. tularensis live vaccine strain (LVS) with a 1,000-fold-smaller dose than an intradermal dose results in similar growth kinetics and peak bacterial burdens. In spite of similar bacterial burdens, we demonstrate a difference in the quality, magnitude, and kinetics of the primary acute T-cell response depending on the route of inoculation. Further, we show that prostaglandin E(2) secretion in the lung is responsible for the difference in the gamma interferon (IFN-gamma) response. Intradermal inoculation led to a large number of IFN-gamma(+) T cells 7 days after infection in both the spleen and the lung. In contrast, intranasal inoculation induced a lower number of IFN-gamma(+) T cells in the spleen and lung but an increased number of Th17 cells in the lung. Intranasal infection also led to a significant increase of prostaglandin E(2) (PGE(2)) in the bronchoalveolar lavage fluid. Inhibition of PGE(2) production with indomethacin treatment resulted in increased numbers of IFN-gamma(+) T cells and decreased bacteremia in the lungs of intranasally inoculated mice. This research illuminates critical differences in acute adaptive immune responses between inhalational and dermal infection with F. tularensis LVS mediated by the innate immune system and PGE(2).

Importance of rhizobia in Agriculture: potential of the commercial inoculants and native strains for improving legume yields in different land-use systems

NASA Astrophysics Data System (ADS)

Lesueur, D.; Atieno, M.; Mathu, S.; Herrmann, L.

2012-04-01

such as cropping systems, N fertilization and application of crop residues affect the genetic diversity of native strains of rhizobia nodulating soybean in Kenya without any inoculation. Results showed that nodulation was not significantly affected by the different factors except N fertilization, regardless the season. Nodule occupancy revealed only 3 main profiles representing 93.6% and 92.5% of all the RFLP profiles obtained from 2008 and 2009 nodules respectively. This suggested a low diversity of native rhizobial strains capable to nodulate the promiscuous variety. The cropping system, Nitrogen and Residue applications didn't increase the diversity of the rhizobia but results indicated an effect on the distribution of the 3 profiles within the nodules of the plants. Within same treatments, significant differences were found between the two seasons in term of strains occupying the nodules. It could be explained by the shorter rainfall received in 2008 compared to 2009. Results suggest that cropping systems and both N and crop residues applications affect more specifically plant growth and grain yields than the diversity of the native rhizobia nodulating promiscuous soybean variety. Our work shows how diverse are the factors influencing the success of the field rhizobial inoculation of legumes.