Influence of microbial consortia on the incidence of grey mold (Botrytis cinerea) in strawberry (Monterey variety)

Botrytis cinerea, the causal agent of grey mold disease, is one of the most destructive pathogens of strawberry crops, both in vegetative development and postharvest. The control of this pathogen is complex due to its aggressiveness and ability to attack and infect various plant tissues and is mainly based on chemical control; however, the incorrect use of pesticides, mainly due to overdosing, causes the presence of traces of these agrochemicals in the fruits, as well as the selection of pathogen resistance to fungicides, making it a risk to human health and the environment. The objective of the study was to use biological regulation strategies, with the application of microbial consortia made up of mycorrhizal fungi, antagonistic bacteria and Trichoderma harzianum, as an alternative for the management of grey mold in strawberry crops (Monterey variety) under field conditions. Treatments T4 (mycorrhizal fungi), T8 (mycorrhizal fungi, antagonistic bacteria and T. harzianum) and T2 (T. harzianum) presented the lowest incidence of the pathogen with 2.6, 3.1 and 3.6 %, respectively, compared to control plants with 16.6%. The influence of all biological treatments on the regulation of B. cinerea was greater than the control.

Botrytis cinerea, el agente causal de la enfermedad del moho gris, es uno de los patógenos más destructivos del cultivo de fresa, tanto en el desarrollo vegetativo como en poscosecha. El control de este patógeno es complejo, debido a su agresividad y capacidad de atacar e infectar diversos tejidos de la planta y se basa, principalmente, en el control químico; sin embargo, el uso incorrecto de plaguicidas, principalmente por sobredosificación, provoca la presencia de trazas de estos agroquímicos en los frutos, así como la selección de resistencia del patógeno a los fungicidas, convirtiéndolo en un riesgo para la salud humana y el ambiente. El objetivo del estudio fue utilizar estrategias de regulación biológica, con la aplicación de consorcios microbianos, conformados por hongos micorrícicos, bacterias antagonistas y Trichoderma harzianum, como alternativa para el manejo del moho gris, en cultivos de fresa (variedad Monterey), en condiciones de campo. Los tratamientos T4 (hongos micorrízicos), T8 (hongos micorrízicos, bacterias antagonistas y T. harzianum) y T2 (T. harzianum) presentaron la menor incidencia del patógeno, con 2,6, 3,1 y 3,6 %, respectivamente, en comparación con las plantas control, con 16,6 %. La influencia de todos los tratamientos biológicos en la regulación de B. cinerea fue mayor respecto al control.

Growth of Schizolobium parahyba var. amazonicum seedlings by strains of Trichoderma spp. under boron rates

Brazil has many important native species, which includes Schizolobium parahyba var. amazonicum (Huber ex Ducke) Barneby, however, the growth of this species is inhibited in soils with low boron contents, or excess boron, which causes phytotoxicity. Applications of strains of Trichoderma spp. increase the plants' tolerance to abiotic stresses, including tolerance to low or excess levels of nutrients such as boron (B). Thus, the objective of this work was to evaluate plant development and the effect of strains of Trichoderma spp. in Schizolobium parahyba var. amazonicum seedlings grown under different boron rates. A randomized block experimental design was used, with a 4×5 factorial scheme, consisting of 4 strains of Trichoderma spp. and 5 B rates (0, 0.5, 1.0, 1.5, and 2.0 mg dm-3), with seven replications, in greenhouse. Plant height, stem diameter, fresh and dry mass, leaf, stem and total were evaluated at 120 days after emergence. The stains of Trichoderma spp. and B rates presented significant interaction for all evaluated variables, decreasing phytotoxicity. The strain IBLF006WP (T. harzianum) showed a higher capacity of increasing the plants' tolerance to boron, followed by URM5911 (T. asperellum). However, the beneficial effect of increasing this tolerance with the application of these strains is only feasible for soils with high contents of this micronutrient in the soil.

Suppression and management of Meloidogyne incognita in soil using Trichoderma harzianum NFCF160 and Trichoderma virens Isf-77

Aims@#Meloidogyne incognita adversely affects numerous crop plants worldwide. Therefore, the modern world has been moving towards biocontrol methods to prevent nematode attacks. This study was aimed to (i) investigate the potential use of Trichoderma harzianum NFCF160 and T. virens Isf-77 in managing M. incognita in soil and (ii) identify trapping mechanisms employed by both Trichoderma strains to suppress M. incognita.@*Methodology and results@#Three weeks old, Basella alba L. plants were subjected to five different treatments. The above and below ground growth parameters and the galling indices of these plants were measured every four weeks for three sampling times. Trapping mechanisms employed by Trichoderma strains were examined following plate assays. Plants treated with T. harzianum NFCF160 and T. virens Isf-77 had significantly higher values for the total number of leaves (34 ± 2.84) and (27 ± 2.61), fresh weight of the shoot (81 ± 9.51 g) and (91 ± 9.70 g), dry weight of the shoot (71 ± 5.24 g) and (62 ± 5.81 g), respectively eight weeks after inoculation of M. incognita. Significantly low galling indices (2 and 2) were recorded in B. alba treated with Trichoderma strains. Both Trichoderma strains exhibited various nematode-trapping mechanisms, such as non-constricting rings and adhesive spores.@*Conclusion, significance and impact of study@#This investigation highlighted the potential of both Trichoderma strains as biocontrol agents to control M. incognita effect in sustainable agriculture.

Evaluation on the effectiveness of combination of biocontrol agents in managing Ganoderma boninense of oil palm

Aims@#The development of an effective biocontrol formulation for inhibition of Ganoderma boninense, a well-known destructive pathogen in oil palm plantation is important to prolong the palm’s lifespan and reduce the losses due to this disease. In this paper, we present some new bioformulations with combination of different types of biocontrol agents in managing basal stem rot (BSR) disease. @*Methodology@#The effectiveness of the treatments designed as T1 (Trichoderma harzianum + Lecanicillium spp. + Streptomyces sundarbansensis + Pseudomonas aeruginosa), T2 (Penicillium simplicissimum + Lecanicillium sp. + S. sundarbansensis + P. aeruginosa), T3 (P. simplicissimum + P. aeruginosa) and T4 (LEStani®) was evaluated through treatment on the oil palm seedlings artificial infected by G. boninense and the results were expressed in disease severity index (DSI), bole severity index (BSI) and ergosterol content.@*Conclusion, significance and impact of study@#All tested treatments (T1-T4) managed to control the severity of the Ganoderma infection from continuously increasing when the treatments were applied either one month before or after artificial inoculation. The disease severity of infected seedlings without treatments had increased for almost 2-fold at the end of the trial. Moreover, T1 had the greatest inhibition of G. boninense with the lowest ergosterol content (a bioindicator of Ganoderma colonization) detected (676.67 g/mL), which is about 1.9-fold lower than control (1273.33 ug/mL) without treatments and a BSI score of 1. Based on the effectiveness among the four tested biocontrol formulations, T1 is the most promising formulation to be further evaluated in the field for control of BSR disease. However, more research is needed in the future to estimate the effective amount for application in open environment.

Antimicrobial activity screening of endophytic fungi extracts isolated from brown algae Padina sp.

This study aims to evaluate the antimicrobial activities of endophytic fungi from marine brown algae Padina sp.collected from Nirwana Beach, Padang, West Sumatera, Indonesia. The isolation of endophytic fungi was conductedusing dilution method with Sabouraud Dextrose Agar (SDA) + Chloramphenicol as a growth medium. Nine fungistrains have been isolated from this alga. Purely isolated fungi were cultivated using rice as a medium at roomtemperature for 3–4 weeks. The secondary metabolite produced by fungi was extracted using ethyl acetate (EtOAc)as a solvent. The antimicrobial activity of EtOAc extracts was tested against Staphylococcus aureus, Escherichiacoli, and Candida albicans by using the agar diffusion method. In this research, nine endophytic fungi were isolatedfrom the brown marine algae Padina sp. The results of antimicrobial activity screening showed that one fungal isolate(Nita3) was selected as the most active against S. aureus, E. coli, and C. albicans with a diameter of inhibition zone of20.98 ± 1.56 mm, 17.98 ± 6.58 mm, and 13.60 ± 0 mm, respectively. This selected fungus was identified molecularlyas Trichoderma harzianum. We conclude that T. harzianum can be a source of antimicrobial compounds. However,continuous research is needed to prove its bioactive action

Antimicrobial activity screening of endophytic fungi extracts isolated from brown algae Padina sp.

This study aims to evaluate the antimicrobial activities of endophytic fungi from marine brown algae Padina sp.collected from Nirwana Beach, Padang, West Sumatera, Indonesia. The isolation of endophytic fungi was conductedusing dilution method with Sabouraud Dextrose Agar (SDA) + Chloramphenicol as a growth medium. Nine fungistrains have been isolated from this alga. Purely isolated fungi were cultivated using rice as a medium at roomtemperature for 3–4 weeks. The secondary metabolite produced by fungi was extracted using ethyl acetate (EtOAc)as a solvent. The antimicrobial activity of EtOAc extracts was tested against Staphylococcus aureus, Escherichiacoli, and Candida albicans by using the agar diffusion method. In this research, nine endophytic fungi were isolatedfrom the brown marine algae Padina sp. The results of antimicrobial activity screening showed that one fungal isolate(Nita3) was selected as the most active against S. aureus, E. coli, and C. albicans with a diameter of inhibition zone of20.98 ± 1.56 mm, 17.98 ± 6.58 mm, and 13.60 ± 0 mm, respectively. This selected fungus was identified molecularlyas Trichoderma harzianum. We conclude that T. harzianum can be a source of antimicrobial compounds. However,continuous research is needed to prove its bioactive action.

Secondary Metabolites of Endophytic Fungus Trichoderma harzianum Isolated from Physalis angulata / 中国实验方剂学杂志

Objective:To study the secondary metabolites of the endophytic fungus Trichoderma harzianum,which was isolated from Physalis angulata. Method:The strains were cultured in a big-scale by oscillating incubator. Then the obtained culture liquid and mycelia were extracted by EtOAC and actone,respectively. The extracts were integrated after recovery of solvents, and then the active secondary metabolites were isolated and purified by comprehensive use of open ODS flash column,sephadex LH-20,HPLC,and LC-MS analysis techniques. Their structures were identified according to their physico-chemical properties and nuclear magnetic resonance (NMR) data. The cytotoxicities of the compounds were determined by methyl thiazolyl tetrazolium(MTT) assay. Result:Ten compounds isolated from T. harzianum were identified as destruxin A2(1),destruxin B2(2),3-isobutyl-pyrrolopiperazine-2,5-dione[cyclo (leu-pro dipeptide)](3),cyclo (Phen-pro) dipeptide(4),cyclonerodiol(5),brevianamide F(6),N-acetyltryptamine(7),9-hydroxyl-(2-methylpropyl) isobutyl phthalate(8),5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone(9),and phenylpropionic acid(10). Compounds 1-10 didn't exhibit significant cytotoxicity to lung cancer cell line A549 in the MTT assay (IC50 ≥ 20 mg·L-1). Conclusion:All compounds were isolated from T. harzianum for the first time.

Integrated Control against Rice Blast and Leaf Spot by Trichoderma harzianum and Two Fungicides

Integrated pest management is performed at intervals of 10 days, during all the season of rice cultivation, by the application of Trichoderma harzianum at a concentration of 108 spores/ml, in alternation with the mancozeb at 1000 ppm against rice blast and rice leaf spot and the pyrazophos at 750 ppm against blast. The assessment of symptoms is performed at the beginning of the panicles appearance, by estimating the incidence of the disease and the symptoms severity. Thus, at the end of treatment programs, the alternation of pyrazophos and T. harzianum reduced blast at a rate similar to that noted when pyrazophos is used alone (i.e. respectively 90.5 and 89.1%). This percentage is better than that recorded following treatment by T. harzianum alone (78.4%). Mancozeb alternated with T. harzianum reduced blast at a rate of (83.49%) compared with the fungicide or the antagonist alone (77 and 78.4%). The application of mancozeb alone reduced the leaf spot at a rate similar to that noted following its alternation with T. harzianum (79.2 and 75.64%) and better than that obtained after treatment with T. harzianum alone (69.5%).

Selectivity of different biological products to the egg parasitoid Telenomus remus (Hymenoptera: Platygastridae)

ABSTRACT The selectivity of five entomopathogens and a chemical insecticide (positive control) to pupae and adults of the egg parasitoid Telenomus remus (Nixon) was evaluated in the laboratory under controlled environmental conditions according to protocols established by the International Organization for Biological Control. Baculovirus anticarsia (Baculovirus AEE®), Bacillus thuringiensis var. kurstaki (Thuricide®), Bacillus thuringiensis var. aizawai (Agree®), Beauveria bassiana (Boveril®), Metarhizium anisopliae (Metarril®) and Trichoderma harzianum (Trichodermil®) are harmless to T. remus pupae, and adults. Thus, our results suggest that the insect control strategies applied here are compatible since entomopathogens were classified as harmless to T. remus in most examined cases and therefore facilitate a joint application to control different pests. Bacillus thuringiensis var. kurstaki (Dipel®), despite being classified as slightly harmful in some of the evaluations, can still be considered compatible for use together with T. remus, especially when compared with chemical insecticides such as chlorpyrifos that might be considered harmful to the parasitoid survival.

Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum / Avaliações das propriedades catalíticas e cinéticas da beta-glucosidase isoladas de um fungo antagonísta altamente eficiente Trichoderma harzianum

Due to the toxicity and inefficiency of chemical fungicides to control infestation of Macrophomina phaseolina (Tassi) Goid which causes charcoal rot in plants, a biotechnological approach using - glucosidase (EC.3.2.1) as the alternative bioactive ingredient in fungicide is hereby, proposed. The extracellular enzyme was isolated from a highly efficient fungal antagonist, Trichoderma harzianum T12. The highly similar molecular masses obtained using SDS-PAGE (96 kDa) and MALDI-TOF mass spectrometry (98.3 kDa) affirmed that the -glucosidase was purified to homogeneity. Consequently, optimum catalytic parameters that rendered the highest enzyme activity were found to be: 45°C, pH 7, inoculum size of 10 % (w/v), supplementation with metal ions Zn2+ and Mn2+ ions, and Tween 80. Addition of wheat bran and (NH4)2SO4 as carbon and nitrogen sources also improved enzyme activity. BLASTn showed the sequence of -glucosidase T12 was highly identical to other -glucosidases viz. T. harzianum strain IOC-3844 (99%), T. gamsii and T. virens bgl1 (86 %) as well as T. reesei strain SJVTR and T. viride strain AS 3.3711 (84 %). Kinetic assessment showed that -glucosidase T12 catalyzes hydrolytic activity is characterized by a Km of 0.79 mM and Vmax of 8.45 mM min-1 mg-1 protein, with a corresponding kcat of 10.69 s-1.

Devido à toxicidade e ineficiência dos fungicidas químicos para controlar a infestação de Macrophomina phaseolina (Tassi) Goid que causa o apodrecimento das plantas, uma abordagem biotecnológica usando - glicosidase (EC.3.2.1) como o ingrediente bioativo alternativo do fungicida é por este meio, proposto. A enzima extracelular foi isolada de um antagonista fúngico altamente eficiente, o Trichoderma harzianum T12. As massas moleculares altamente similares obtidas usando SDS-PAGE (96 kDa) e espectrometria de massa MALDI-TOF (98,3 kDa) afirmaram que a -glicosidase foi purificada até a homogeneidade. Consequentemente, os parâmetros catalíticos ótimos que apresentaram a maior atividade enzimática foram: 45°C, pH 7, tamanho do inóculo de 10% (p / v), suplementação com íons de metais Zn2+ e Mn2+, e Tween 80. Adição de farelo de trigo e (NH4) 2SO4 como fontes de carbono e nitrogênio também melhoraram a atividade enzimática. O BLASTn mostrou que a sequência da -glicosidase T12 era altamente idêntica a outras -glicosidase viz. A estirpe T. harzianum IOC-3844 (99%), T. gamsii e T. virens bgl1 (86%) assim como a estirpe T. reesei SJVTR e a estirpe T. viride AS 3.3711 (84%). A avaliação cinética mostrou que -glicosidase T12 catalisa a actividade hidrolítica caracterizada por um Km de 0,79 mM e Vmax de 8,45 mM min-1 mg-1 de proteína, com um correspondente kcat de 10,69 s-1.

New Report of Three Unrecorded Species in Trichoderma harzianum Species Complex in Korea

The genus Trichoderma (Hypocreaceae, Ascomycota) consists of globally distributed fungi. Among them, T. harzianum, one of the most commonly collected Trichoderma species, had been known as a polyphyletic or aggregate species. However, a total of 19 species were determined from the polyphyletic groups of T. harzianum. Thus, we explored Korean “T. harzianum” specimens that were collected in 2013–2014. These specimens were re-examined based on a recent study with translate elongation factor 1-alpha (EF1α) sequences to reveal cryptic Trichoderma species in Korea. As a result, four different species, T. afroharzianum, T. atrobruneum, T. pyramidale, and T. harzianum, were identified. Except T. harzianum, the other three species have not been reported in Korea. In this work, we describe these species and provide figures.

Laboratory efficacy of mycoparasitic fungi (Aspergillus tubingensis and Trichoderma harzianum) against tropical bed bugs (Cimex hemipterus) (Hemiptera: Cimicidae)

Objective:To test the effectiveness of conidial spore formulations [Aspergillus tubingensis (A.tubingensis) and Trichoderma harzianum (T.harzianum)] against tropical bed bugs,Clnex hemipterus.Methods:Spore formulations were made from two fungal strains,T.harzianum and A.tubingensis.The bed bugs were exposed to the conidial spores placed soaked onto a fabric cloth for 1 h and the mortality counts were recorded daily until 14 days.Results:Mean survival times based on Kaplan-Meier survival analysis showed no significant differences between all the concentrations in both the fungal isolates:T.harzianum and A.tubingensis.However,the evaluation of both the isolates in terms of virulence resulted in low lethal hours in all the concentrations except for the high concentration ofA.tubingensis (LT50 =44.629 h) at the conidial exposure of 1 × 106 spores/mL.Rapid mortality of the bed bugs was observed from Day 6 to Day 12,ranging from 13％ to 90％ in all three concentrations of A.tubingensis.With reference to the T.harzianum exposure,the concentration of 1 × 104 spores/mL displayed a gradual increase in the percentage mortality of 90 on Day 14.Conclusions:Approaches to the bed bugs treatment should be explored in-depth using a natural biological agent like fungus especially A.tubingensis to reduce this pest population,in order to replace chemical methods.

Optimization of Fungal Enzyme Production by Trichoderma harzianum KUC1716 through Surfactant-Induced Morphological Changes

The morphological optimization of Trichoderma harzianum was carried out using several surfactants to achieve increased cellulase production. Addition of the surfactants to the culture medium successfully modified the fungal morphology from an aggregated form to a dispersed form. Optimization of the fungal morphology increased cellulase activity up to 177%. The morphologically optimized conditions enhanced the accessibility of the fungus to substrates and thus promoted cellulase production.

Laboratory efficacy of mycoparasitic fungi (Aspergillus tubingensis and Trichoderma harzianum) against tropical bed bugs (Cimex hemipterus) (Hemiptera: Cimicidae)

Objective To test the effectiveness of conidial spore formulations [Aspergillus tubingensis (A. tubingensis) and Trichoderma harzianum (T. harzianum)] against tropical bed bugs, Cimex hemipterus. Methods Spore formulations were made from two fungal strains, T. harzianum and A. tubingensis. The bed bugs were exposed to the conidial spores placed soaked onto a fabric cloth for 1 h and the mortality counts were recorded daily until 14 days. Results Mean survival times based on Kaplan–Meier survival analysis showed no significant differences between all the concentrations in both the fungal isolates: T. harzianum and A. tubingensis. However, the evaluation of both the isolates in terms of virulence resulted in low lethal hours in all the concentrations except for the high concentration of A. tubingensis (LT

The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens

Abstract The antagonistic potential of Trichoderma strains was assayed by studying the effect of their culture filtrate on the radial growth of Sclerotium rolfsii, the causal agent of chickpea collar rot. Trichoderma harzianum-1432 (42.2%) and Trichoderma atroviride (40.3%) were found to be strong antagonists. To enhance their antagonistic potential, mutagenesis of these two selected strains was performed. Two mutants, Th-m1 and T. atroviride m1, were found to be more effective than their parent strains. The enzymatic activities of the selected parent and mutant strains were assayed, and although both mutants were found to have enhanced enzymatic activities compared to their respective parent strains, Th-m1 possessed the maximum cellulase (5.69 U/mL) and β-1,3-glucanase activity (61.9 U/mL). Th-m1 also showed high competitive saprophytic ability (CSA) among all of the selected parent and mutant strains, and during field experiments, Th-m1 was found to successfully possess enhanced disease control (82.9%).

Chemical and biological control of Sclerotinia stem rot in the soybean crop / Controle químico e biológico da podridão branca da haste na cultura da soja

It was evaluated the effect of fungicides and the microbial control agent Trichoderma harzianum on the inhibition of the carpogenic and ascospore germination of Sclerotinia sclerotiorum. This study also evaluated the chemical, fungicidal and microbial control of white mold or Sclerotinia stem rot of soybean in the field. Three experiments were conducted, as follows: 1) inhibition of carpogenic germination of sclerotia, 2) inhibition of ascospore germination, and 3) control of Sclerotinia stem rot in a soybean crop under field conditions. The treatments evaluated were fluazinam, procymidone, iprodione, thiophanate-methyl, carbendazim, benzalkonium chloride + fluazinam, and T. harzianum. Procymidone resulted in an inhibition of 13.5% and benzalkonium chloride in an inhibition of 13.9% in an ascospore germination test. Fluazinam and procymidone were the most effective in reducing the production of ascospores/apothecium, representing 65.6% and 82.4% of inhibition. Procymidone and fluazinam if combined or not with benzalkonium chloride were the most effective in controlling sclerotinia stem rot under field conditions when applied at the onset of flowering and 15 days later. In the 2009-10 harvest, these two fungicides reduced the incidence of Sclerotinia stem rot by 73.1 and 71.6% and in the 2010-11 harvest by 75.7 and 77.6%, respectively.

Avaliaram-se o efeito de fungicidas e agente de controle biológico Trichoderma harzianum na inibição da germinação carpogênica e de ascósporos de Sclerotinia sclerotiorum. Também foram avaliados os controles químico e biológico do mofo branco ou podridão branca da haste na cultura da soja em condições de campo. Foram desenvolvidos três ensaios, sendo: 1) inibição da germinação carpogênica; 2) inibição da germinação de ascósporos; e 3) controle da podridão branca da haste na cultura da soja em condições de campo. Os tratamentos avaliados foram fluazinam, procimidona, iprodione, tiofanato metílico, carbendazim, cloreto de benzalcônio + fluazinam e T. harzianum. O procimidona resultou em inibição de 13,5% e o cloreto de benzalcônio + fluazinam em 13,9% de inibição no teste de inibição da germinação dos ascósporos. Fluazinam e procimidona foram os mais eficientes na redução da produção de ascósporos/apotécio representando 65,6% e 82,4% de inibição. O procimidona e fluazinam se combinados com cloreto de benzalcônio ou não foram os mais eficientes no controle da podridão branca da haste em condições de campo, quando aplicados no início do florescimento e 15 dias após. Na safra 2009-10, esses dois fungicidas reduziram a incidência da podridão branca da haste, em 73,1 e 71,6% e, na safra 2010-11, reduziram em 75,7 e 77,6%, respectivamente.

Evaluation of Biological Efficacy of Trichoderma Species Isolates Against Alternaria Leaf Spot Disease of Sesame.

Alternaria leaf spot disease is a major threat to sesame (Sesamum orientale L.) caused by Alternari asesami. Induced resistance is an alternative to systemic disease resistance response of plants. The present study aims to evaluate Trichoderma species efficacy as biocontrol via induction of resistance against A. sesami in sesame species. During in vitro bio control test, T. harzianum colonize and parallely inhibit the growth of the fungal pathogen. Expression of various defence related enzymes observed in sesame induce resistance against the pathogen infection in the host. T. harzianum coupled with inoculation of A. sesami enhance the remarkable induction of defence enzyme such as peroxidase (POX), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL) and also the phenolic content compared with the control. The enzyme activity increased from 48 h of sampling and peaked at 72 h and then decreased after 72 h. In greenhouse and field experiments, soil treatment with a powder formulation of T. harzianum two weeks before planting or at the time of planting reduced significantly the incidence of diseases on both the wild and cultivar Thilarani.The results demonstrate that T. harzianumcan be successfully applied as a biological control against Alternaria leaf spot disease in sesame.

Susceptibility of Malaysian rice varieties to Fusarium fujikuroi and in vitro activity of Trichoderma harzianum as biocontrol agent

Aims: Bakanae disease on rice has been widely distributed in all countries where rice is grown commercially, especially in Asian countries including Malaysia. As an alternative measure in controlling Fusarium fujikuroi, two approaches have to be adapted i.e. by using resistant varieties and biocontrol agents as reported in the present study. Methodology and results: A total of 31 Malaysian rice varieties were used in screening and results showed that variety MR211 was the most susceptible and MR220 was slightly susceptible. Out of 60 isolates of Trichoderma harzianum isolated from soils in Malaysia and tested against the pathogen under in vitro condition, 13 isolates showed high percentage of inhibition (PIRG > 60%). All isolates of T. harzianum showed that the PIRGs were significantly different at p≤0.05 with those of control plates. Conclusion, significance and impact of study: Biocontrol agent and resistant variety are better alternative for controlling plant diseases. We found a variety MR220 was slightly susceptible, but none of tested varieties is resistant towards pathogen of bakanae disease. T. harzianum has the ability to inhibit the growth of F. fujikuroi (T3068P) under in vitro condition. The findings of the Malaysian susceptible/resistant variety and potential T. harzianum isolate as a biocontrol agent of bakanae are important for future tests in the plant house and field trials.

Manejo da podridão-de-Sclerotium em pimentão em um argisolo no Amazonas / Southern blight management of bell pepper in an argissol in the Amazonas State

Southern blight is a common disease in plants belonging to the Solanaceae family in the Amazon. To compare southern blight management strategies for bell pepper (Capsicum annuum, L. Solanaceae) a field experiment was conducted in a split plot design with main plots arranged in randomized blocks, with six replicates for each treatment, in a red-yellow Argisol (Ultisol) artificially infested with Sclerotium rolfsii. The main factor was soil cover (sawdust mulch or bare soil) The secondary factors were the incorporation to the soil: 1) compost (3 L by planting hole), 2) rice colonized by Trichoderma harzianum (90 g by planting hole, with ≈ 1.4 x 10(9) conidia g-1) and 3) compost and T. harzianum in the same proportion described previously; 4) control. All plants were fertilized with 1.5 L of compost by planting hole except those in the treatments with 3 L of compost by hole. The main plot had three rows with ten bell pepper plants (0.50 x 1.0 m) and the subplot, three rows with five plants. Southern blight incidence was accessed twice a week. The sawdust mulch increased the disease incidence significantly. In the mulched plots the disease intensity, expressed as the area under disease progress curve (AUDPC), increase 35.5% compared with bare soil. Trichoderma harzianum or the increase in the amount of compost (1.5 L to 3 L by planting hole) reduced the AUDPC by 38.1% and of 37.5%, respectively. Furthermore T. harzianum or the increase in the amount of compost, even with sawdust mulch added, reduced the AUDPC by 52.8% and in 55.1%, respectively, in comparison with mulch alone. These results suggested that the utilization of T. harzianum or the increase in the amount of compost by planting hole are viable strategies to southern blight management in bell pepper. The sawdust surface mulch is not recommended in an S. rolfsii infested area.

A podridão-de-Sclerotium é uma doença comum em plantas da família Solanaceae na Amazônia. Visando avaliar estratégias de manejo para esta doença em pimentão (Capsicum annuum, L. Solanaceae), foi conduzido experimento em campo em blocos casualizados com parcelas subdivididas e seis repetições, em Argissolo Vermelho-Amarelo artificialmente infestado com Sclerotium rolfsii. O tratamento principal foi a cobertura do solo (cobertura do solo com serragem ou solo nu). Os tratamentos secundários consistiram na adição ao solo de: 1) composto vegetal (3 L por cova), 2) arroz colonizado com Trichoderma harzianum (90 g por cova contendo ≈ 1,4 x 10(9) conídios g-1), 3) composto vegetal e T. harzianum nas mesmas proporções descritas anteriormente e 4) testemunha. Todas as plantas receberam apenas adubação orgânica com composto vegetal na proporção de 1,5 L por cova, exceto as dos tratamentos com 3 L de composto por cova. A parcela principal foi constituída de três fileiras com dez plantas de pimentão (0,50 x 1,0 m) e cada subparcela continha três fileiras com cinco plantas. A incidência da podridão-de-Sclerotium foi avaliada duas vezes por semana. A cobertura morta favoreceu significativamente a ocorrência da doença. Nas parcelas com esse tratamento o aumento da intensidade da doença, expressa em área abaixo da curva de progresso da doença (AACPD), foi 35,5% maior, em comparação com as parcelas sem cobertura morta. A aplicação de T. harzianum ou o incremento na quantidade de composto (de 1,5 para 3 L por cova) reduziu a AACPD em 38,1% e 37,5%, respectivamente. A aplicação de T. harzianum ou o incremento na quantidade de composto, mesmo nos tratamentos com cobertura morta, reduziu significativamente a AACPD em 52,8% e em 55,1%, respectivamente, em comparação com o tratamento apenas com cobertura morta. Esses resultados sugerem que a utilização de T. harzianum e o aumento na quantidade de composto por cova são estratégias eficientes de manejo da podridão-de-Sclerotium em...

Sugarcane bagasse as feedstock for cellulase production by Trichoderma harzianum in optimized culture medium

This work aimed at the production of cellulases from pretreated sugarcane bagasse by the filamentous fungus Trichoderma harzianum IOC 3844 and their application in the hydrolysis of this same substrate, for a future use in second-generation ethanol production. The production of cellulases was optimized, which resulted in high enzymatic activities after 42 hrs of process in an instrumented bioreactor (CMCase 27,017 U x L-1; FPase 1,225 U x L-1; and β-glucosidase 609 U x L-1). The enzymatic extract was concentrated by using a hollow fiber membrane filtration system. The concentrated extract was applied in the hydrolysis of pretreated sugarcane bagasse, after 28 hrs of enzymatic reaction, displaying a similar catalytic performance of that attained with a commercial enzymatic preparation (hydrolysis efficiency of roughly 50%). Finally, the enzymatic extract was partially characterized in terms of the molecular weights of the main activities of the enzymatic pool. Electrophoretic analysis identified eleven protein bands; six of them were related to CMCase activity and revealing molecular weights that varied from 48 to 78 kDa, and two bands were associated with β-glucosidase activity and having molecular weights of 75 and 85 kDa.