Thirteen new peptaibols with antimicrobial activities from Trichoderma sp / 中国天然药物

From the fungus Trichoderma sp., we isolated seven novel 18-residue peptaibols, neoatroviridins E-K (1-7), and six new 14-residue peptaibols, harzianins NPDG J-O (8-13). Additionally, four previously characterized 18-residue peptaibols neoatroviridins A-D (14-17) were also identified. The structural configurations of the newly identified peptaibols (1-13) were determined by comprehensive nuclear magnetic resonance (NMR) and high-resolution electrospray ionization tandem mass spectrometry (HR-ESI-MS/MS) data. Their absolute configurations were further determined using Marfey's method. Notably, compounds 12 and 13 represent the first 14-residue peptaibols containing an acidic amino acid residue. In antimicrobial assessments, all 18-residue peptaibols (1-7, 14-17) exhibited moderate inhibitory activities against Staphylococcus aureus 209P, with minimum inhibitory concentration (MIC) values ranging from 8-32 μg·mL-1. Moreover, compound 9 exhibited moderate inhibitory effect on Candida albicans FIM709, with a MIC value of 16 μg·mL-1.

Trichoderma asperellum isolated from salinity soil using rice straw waste as biocontrol agent for cowpea plant pathogens.

This study discusses isolation and identification new fungal isolate from salinity soil for controlling soil borne diseases. Among sixteen fungal, a potent isolate coded SRBP_ZSHSG1 was isolated from Sugar beet rhizospher samples collected from Al-Hosainia localities-El-Sharkia-Egypt. Traditional methods consistent with phylogenetic analysis of 18S rRNA sequences showed SRBP_ZSHSG1 has 100% similarity with Trichoderma strains and the most closest is Trichoderma asperellum. Thus, it proposed name Trichoderma asperellum SRBP_ZSHSG1 (ID: KP336489). Results proved SRBP_ZSHSG1 followed by T. roseum and Chaetomium globosum were highly inhibitors to the tested pathogens. These results were confirmed by field experiments. SRBP_ZSHSG1 was able to grow on rice straw (biostraw) and produce most active compounds. The biostaw extract was the most effective bioagent and recorded highest reduction in pathogen numbers. GC/MS analysis of ethyl acetate extract revealed the presence of 9 compounds. These compounds were determined 4 volatile alcohols (1-4) and fatty acid esters (5-9).

Seasonal effects and antifungal activity from bark chemical constituents of Sterculia apetala (Malvaceae) at Pantanal of Miranda, Mato Grosso do Sul, Brazil / Efeitos sazonais e atividade antifúngica dos constituintes químicos das cascas de Sterculia apetala (Malvaceae) no Pantanal de Miranda, Mato Grosso do Sul, Brasil

In the Southern Pantanal, the hyacinth macaw (Anodorhynchus hyacinthinus), an endangered species, often chooses the manduvi tree (Sterculia apetala) as a nesting site, because of its physical properties. In addition, the chemical composition of the wood may also contribute to a nesting selection by the hyacinth macaws. The objective of this study was to determine the main chemical components of S. apetala bark for two seasons, and evaluate its fungicidal potential. Bark samples from S. apetala trees with and without nests of A. hyacinthinus were collected in January (wet season) and August (dry season) of 2012. The inhibition of mycelium growth (MGI) from tree samples with and without nests were assessed using a phytochemical analysis to evaluate their antifungal activity against Trichoderma sp. Phytochemical analysis confirmed the presence of phenolic compounds and flavonoids. In both seasons, samples obtained from nested trees had higher content of total phenols than those collected from non-nested trees. The average content of total flavonoids was higher in January for samples with nest and in August for samples without nest. All selected samples showed antifungal activity, and those with nest collected in August (peak of hyacinth macaw breeding) resulted in an MGI of 51.3%. Therefore, this percentage, related to the content of flavonoids and the presence of coumarins, may influence the reproductive success of hyacinth macaws and other species of birds, in this region. This is the first chemical study report with the stem bark of S. apetala.

No Pantanal Sul, as araras-azuis (Anodorhynchus hyacinthinus), espécie em extinção, escolhem preferencialmente o manduvi (Sterculia apetala) como local de nidificação devido principalmente às características físicas da árvore. Porém, a composição química da madeira também pode interferir nesta seleção. Os objetivos deste trabalho foram determinar as principais classes de substâncias químicas presentes nas cascas de S. apetala em duas épocas do ano e seu potencial antifúngico. As cascas de árvores, com e sem a presença de ninhos de A. hyacinthinus, foram coletadas em janeiro (período de chuvas) e agosto (período de seca) de 2012. Por meio das análises fitoquímicas e quantificação de flavonóides, foram selecionadas amostras de uma árvore com ninho e outra sem ninho para determinar seu potencial antifúngico frente à Trichoderma sp. através da porcentagem de inibição do crescimento micelial (PIC). A investigação fitoquímica do extrato etanólico das cascas revelou a predominância de compostos fenólicos e flavonóides. O teor médio de fenóis totais foi superior para as amostras com ninho, em relação às sem ninho, nos dois períodos de coleta e, para flavonóides, os valores foram superiores em janeiro para a amostra com ninho e em agosto, para sem ninho. As amostras selecionadas apresentaram potencial antifúngico, sendo que com ninho, coletada em agosto (auge do período de reprodução das araras-azuis), resultou em um PIC de 51,3%, valor relacionado aos flavonóides e as cumarinas, fator que pode influenciar o sucesso reprodutivo da arara azul e outras espécies de aves nesta região. Este trabalho foi o primeiro a identificar os componentes químicos da casca do tronco de S. apetala.

Efficiency of treatments for controlling Trichoderma spp during spawning in cultivation of lignicolous mushrooms

Trichoderma spp is the cause of the green mold disease in mushroom cultivation production. Many disinfection treatments are commonly applied to lignocellulose substrates to prevent contamination. Mushroom growers are usually worried about the contaminations that may occur after these treatments during handling or spawning. The aim of this paper is to estimate the growth of the green mold Trichoderma sp on lignocellulose substrates after different disinfection treatments to know which of them is more effective to avoid contamination during spawning phase. Three different treatments were assayed: sterilization (121 ºC), immersion in hot water (60 and 80 ºC), and immersion in alkalinized water. Wheat straw, wheat seeds and Eucalyptus or Populus sawdust were used separately as substrates. After the disinfection treatments, bagged substrates were sprayed with 3 mL of suspension of conidia of Trichoderma sp (10(5) conidia/mL) and then separately spawned with Pleurotus ostreatus or Gymnopilus pampeanus. The growth of Trichoderma sp was evaluated based on a qualitative scale. Trichoderma sp could not grow on non-sterilized substrates. Immersions in hot water treatments and immersion in alkalinized water were also unfavorable treatments for its growth. Co- cultivation with mushrooms favored Trichoderma sp growth. Mushroom cultivation disinfection treatments of lignocellulose substrates influence on the growth of Trichoderma sp when contaminations occur during spawning phase. The immersion in hot water at 60 ºC for 30 min or in alkalinized water for 36 h, are treatments which better reduced the contaminations with Trichoderma sp during spawning phase for the cultivation of lignicolous species.

Bioethanol production from sweet potato (Ipomoea batatas L.) flour using co-culture of Trichoderma sp. and Saccharomyces cerevisiae in solid-state fermentation

The aim of this work was to study the optimiation of co-culturing of Trichoderma sp. and Saccharomyces cerevisiae (1:4 ratio) on sweet potato (Ipomoea batatas L.) flour (SPF) for the production of bio-ethanol in solid-state fermentation (SSF). Maximum ethanol (172 g/kg substrate) was produced in a medium containing 80% moisture, ammonium sulphate 0.2%, pH 5.0, inoculuted with 10% inoculum size and fermented at 30ºC for 72h. .Concomitant with highest ethanol concentration, maximum ethanol productivity (2.8 g/kg substrate/h), microbial biomass (23×10(8) CFU/ g substrate), ethanol yield (47 g/100g sugar consumed) and fermentation efficiency (72%) were also obtained under these conditions. Cell interaction was observed familiar between the viable cells of Trichoderma sp. and S. cerevisiae when co-cultured. Ethanol production ability by the co-culture was 65 % higher than the single culture of S. cerevisiae from un-saccharified SPF.

Evaluación de la estabilidad de Trichoderma sp. y Azotobacter sp. conservados por diferentes métodos / Evaluation of the stability of Trichoderma sp. and Azotobacter sp. preserved by different methods

En el Laboratorio de Microbiología Aplicada de la Universidad Francisco de Paula Santander (Cúcuta, Colombia) constantemente ingresan al Banco de Cepas, cultivos microbianos de interés biotecnológico, especialmente del sector agrícola, los cuales son utilizados en las actividades de docencia e investigación. Por esta razón, existe el interés de mantener viables a través del tiempo estos cultivos microbianos, para lo cual se realizó en esta investigación, la evaluación de la estabilidad de los aislados de Trichoderma sp. y Azotobacter sp., utilizando las técnicas de conservación en viales con solución salina estéril (0,85% NaCl) en refrigeración (4°C) y temperatura ambiente (30°C), suelo estéril en refrigeración (4°C), comparados con la metodología de repiques sucesivos como tratamiento control. Los resultados no mostraron diferencias significativas según el Test de Duncan (P≤0,05) en la tasa de supervivencia microbiana entre los métodos de conservación. Sin embargo, se observó en las técnicas de viales con solución salina estéril y suelo estéril mantenidos en refrigeración, mayor estabilidad en la concentración celular durante los cuatros meses de evaluación, sin registrar contaminación en los cultivos. Así mismo, se registró un óptimo crecimiento macroscópico de los cultivos microbianos y sus características microscópicas se mantuvieron estables en estos dos métodos de conservación. Por esta razón, se seleccionaron como técnicas de conservación de estos microorganismos, teniendo en cuenta además, ventajas como la facilidad de la técnica, disponibilidad de equipos, materiales y personal con los que se cuenta en el laboratorio.

In the Laboratory of Applied Microbiology of the University Francisco de Paula Santander (Cúcuta, Colombia), strains of microbial cultures of biotechnological interest are constantly entering, particularly from the agricultural sector, which are used for teaching and research activities. There is an interest in keeping them viable over time, which was the main reason of this research: To evaluate the stability of the isolates of Trichoderma sp. and Azotobacter sp., using different conservation techniques: vials with saline solution (0,85% NaCl) in two temperature conditions: refrigeration (4°C) and room temperature (30°C), sterile soil under refrigeration (4°C), and successive peals methodology as the control treatment. Results showed no significant differences according to Duncan test (P ≤ 0.05) in the microbial survival ratio between preservation methods. However, a greater stability in cell concentration was observed with the techniques of vials with sterile saline solution and sterile soil -both under refrigeration-, during the four-month evaluation, with no register of contamination on cultures. Also, there was an optimum growth of microbial cultures macroscopically and the microscopic characteristics were stable in these two methods of preservation for both strains. For this reason, we suggest these conservation techniques for the two microorganisms, considering further advantages such as ease of technology, availability of equipment, materials and personnel available in the laboratory.

Molecular identification and biodegradation of 3-chloropropionic acid (3CP) by filamentous fungi-Mucor and Trichoderma species isolated from UTM agricultural land

Aims: This study was carried out to further characterize fungal species that could degrade 3-chloropropionic acid (3CP) as sole source of carbon and energy. Methodology and Results: Both fungi were able to grow on 3CP after 10 days on solid minimal media. Based on sequencing of its segment of 18S rRNA these isolates were identified as Mucor sp. SP1 and Trichoderma sp. SP2. The isolated strains were not able to grow on media plates containing 10 mM of 2,2-dichloropropionate (2,2DCP) as sole source of carbon. 3CP degradation was observed in liquid minimal medium containing 10 mM 3CP after 18 days culture period. The chloride ion released was detected in both growth medium containing Mucor sp. SP1 and Trichoderma sp. SP2. At least 80% of 10 mM 3CP was utilized in the growth medium. Conclusion, significance and impact of study: Dehalogenase enzyme that can degrade α-chloro-substituted haloalkanoic acids for example 2,2DCP is well studied up to protein crystallization. Very few reports on the degradation of β-chloro-substituted haloalkanoic acids such as 3CP and none from fungi. This study is considered important because it can be compared to that of well-documented α-chloro-substituted haloalkanoic acids degradation. This is the first study to indicate fungal growth on 3CP as sole carbon and energy sources.

EVALUACIÓN DEL ANTAGONISMO Y MULTIPLICACIÓN DE Trichodermasp. EN SUSTRATO DE PLÁTANO EN MEDIO LÍQUIDO ESTáTICO / Evaluation Of Antagonism And Multiplication The Trichodermasp. In The Middle Static Liquid Banana Substrate

Se evaluó la capacidad antagónica de nueve aislados del hongo Trichoderma sp. proveniente de la colección de cepas del Laboratorio de Fitopatología de la Universidad de Córdoba, Colombia y su producción de esporas en medios de cultivo líquido estático, a través de pruebas in vitro de antagonismo, utilizando dos hongos fitopatógenos: Fusarium sp. y Rhizoctonia sp.; esta prueba permitió seleccionar el aislado con la mayor actividad antagónica, para así ajustar la producción de esporas mediante un proceso de fermentación en estado líquido en condición estática mediante procesos de buferización y sin buferización con fosfato 0,2N, utilizándose como sustrato principal harina de plátano (HP) 5%, además de complementos nutricionales que constituyeron los tratamientos: HPM (solución de melaza 5%), HPL (solución de levadura granulada al 2%), HPU (solución de urea 0,1%), y como testigo PGL (papa glucosa líquida). En los ensayos de antagonismo se seleccionó el aislado C2 de Trichoderma sp. por presentar la mayor capacidad antagónica frente al hongo Fusarium sp., con un porcentaje de inhibición del 78,30% y reportando un índice 4 de antagonismo, en los procesos de producción de esporas el tratamiento HPL (harina de plátano más levadura sin buffer) se destacó por su favorable producción de conidias, con una concentración de 1,1 x 10(9) conidias/mL. Los procesos de buferización se diferenciaron significativamente, demostrando que la variable buffer influyó en la producción de conidias, los medios que se amortiguaron mostraron una producción de 1,01 x 10(9) conidias/mL, con respecto a los que no se amortiguaron 6,3 x 10(8) conidias/mL.

The antagonistisc ability of nine isolated natives fungi Trichoderma sp from the collection of strains at Laboratory of phytopathology at University of Cordoba, Colombia was assessed so as its production of spores in crop liquid static medium, cross the in vitro proof antagonism using two phytopathogenic fungi: Fusarium sp. and Rhizoctonia sp., this proof let select the isolated with the major antagonistic activity and adjust the production of spores through fermentation process in liquid static by means of buffer and without buffer with phosphate 0,2 N, being used as principal substrate plantain flour (HP) 5%, besides of nutritional supplements that formed the trataments: HPM (molasses solution 5%), HPL (granulated yeast solution at 2%), HPU (urea solution 0.1%), and as witness PGL (potato glucose liquid). In the test of antagonism was selected the isolated C2 of Trichoderma sp. by present the major antagonistic ability in front of the fungi Fusarium sp., with a inhibition percentage at 78,30% and reporting a index 4 of antagonism, in the process of spores production, the tratament HPL (plantain flour plus yeast without buffer) stressed for the favorable production of conidias, with a concentration of 1,1 x 10(9) conidias/mL. The buffer process were significant differentiated, showing that the variable buffer influenced in the production of conidias, the mediums that shocked in the production show 1,01 x 10(9) conidias/mL, Ïwith respect at don’t shocked 6,3 x 10(8) conidias/mL.

Effect of Fermented Sawdust on Pleurotus Spawn

A method of spawn making procedures through the application of fermented sawdust for the purpose of avoiding contamination by undesirable fungi in the course of Pleurotus ostreatus mycelial growth was evaluated. Of three kinds of supplements, rice bran was the most effective to raise temperature up to 70degrees C. Mycelial activity and density was more considerably improved in the case of using fermented sawdust supplemented with rice bran than the case of non-fermented sawdust. Primordia of Pleurotus ostreatus were formed on fermented sawdust. The substrate of fermented sawdust showed potential to prevent the growth of Trichoderma sp. which caused a symptom on mushroom mycelium, whereas there was nothing to inhibit the growth of Trichoderma sp. during 30 days after inoculation in non-fermented sawdust.