scholarly journals Evaluación de la capacidad antagónica in vitro de cepas de Trichoderma spp frente a Phytophthora cinnamomi, fitopatógeno de Persea americana (Palta)

2020 ◽  
Vol 23 (1) ◽  
pp. 65-70
Author(s):  
Ebeling M. Morales T. ◽  
Mirko D. Lino N. ◽  
Eddy Ortega R. ◽  
Pedro L. Castellanos S.
Keyword(s):  
Phytophthora Cinnamomi ◽  
Persea Americana ◽  
Trichoderma Asperellum ◽  
Trichoderma Spp

Phytophthora cinnamomi es un fitopatógeno reportado como el agente causal de la pudrición radicular del palto en el Perú. En este estudio se evaluó la capacidad antagónica de cepas nativas de Trichoderma spp aisladas de la rizósfera de un cultivo de paltos del distrito de Quilmaná, Cañete. Para la evaluación in vitro de la capacidad antagónica de las cepas aisladas se utilizó la técnica de cultivo dual en donde se midió los porcentajes de inhibición del crecimiento radial (PICR) de P. cinnamomi. Del total de cepas aisladas, 3 (P1.1, P4.9 y P5.3) presentaron mayores PICR con 53.86, 56,24 y 60,49 % respectivamente. La identificación molecular indicó que la cepas P4.9 y P1.1 presentan un 99,66 y 100% de similitud con Trichoderma asperellum mientras que la cepa P5.3 un 98,17% de similitud con Trichoderma spp, según el análisis BLAST. En base a los resultados obtenidos estas cepas de Trichoderma asperellum representan un potencial para ser utilizadas como agentes biocontroladores de P. cinnamomi. por lo que se recomienda realizar ensayo en campo.

scholarly journals Control of root rot (Phytophthora cinnamomi) in avocado (Persea Americana) with bioagents

2020 ◽  
Vol 46 (3) ◽  
pp. 205-211
Author(s):  
Ciro Hideki Sumida ◽  
Lucas Henrique Fantin ◽  
Karla Braga ◽  
Marcelo Giovanetti Canteri ◽  
Martin Homechin
Keyword(s):  
Biological Control ◽  
Root Rot ◽  
Phytophthora Cinnamomi ◽  
Persea Americana ◽  
Field Conditions ◽  
In Vitro Tests ◽  
Trichoderma Spp ◽  
Maize Cultivation ◽  
Antagonistic Potential

ABSTRACT Despite the favorable edaphoclimatic conditions for avocado production in Brazil, diseases such as root rot caused by the pathogen Phytophthora cinnamomi compromise the crop. With the aim of managing root rot in avocado, the present study aimed to evaluate chemical and biological control with isolates of Trichoderma spp. and Pseudomonas fluorescens. Thus, three assays were conducted to assess: (i) mycelial inhibition of P. cinnamomi by isolates of Trichoderma spp. and P. fluorescens from different crop systems; (ii) effect of autoclaved and non-autoclaved metabolites of P. fluorescens, and (iii) chemical or biological treatment of avocado seedlings on the control of root rot under field conditions. The isolates of Trichoderma spp. from maize cultivation soil and the commercial products formulated with Trichoderma presented greater antagonism (p <0.05) to the pathogen P. cinnamomi in the in vitro tests. Similarly, non-autoclaved metabolites of P. fluorescens presented antagonistic potential to control P. cinnamomi. Under field conditions, the fungicide metalaxyl and the bioagents showed effectiveness in controlling P. cinnamomi, as well as greater root length and mass. Results demonstrated potential for the biological control of avocado root rot with Trichoderma spp. and P. fluorescens.

scholarly journals Endophytic Trichoderma Species Isolated from Persea americana and Cinnamomum verum Roots Reduce Symptoms Caused by Phytophthora cinnamomi in Avocado

Plants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1220
Author(s):  
Petra Andrade-Hoyos ◽  
Hilda Victoria Silva-Rojas ◽  
Omar Romero-Arenas
Keyword(s):  
Root Rot ◽  
Phytophthora Cinnamomi ◽  
Disease Incidence ◽  
Persea Americana ◽  
Dual Culture ◽  
Internal Transcribed Spacer Region ◽  
Trichoderma Spp ◽  
Trichoderma Species ◽  
Potato Dextrose Agar Medium

Avocado root rot caused by the oomycete Phytophthora cinnamomi is a severe disease that affects avocado production in Mexico and worldwide. The use of biological control agents such as Trichoderma species isolated from places where the disease is always present, represents an efficient alternative to reduce losses. Thus, the objective of this research was to evaluate the biocontrol ability of 10 endophytic Trichoderma spp. strains against P. cinnamomi tested both in vitro and in the greenhouse. The endophytic Trichoderma spp. were recovered from Persea americana and Cinnamomum verum roots, isolated and purified on potato–dextrose–agar medium. Ten strains were identified by phylogenetic reconstruction of the internal transcribed spacer region of rDNA sequences as T. asperellum (T-AS1, T-AS2, T-AS6, and T-AS7), T. harzianum (T-H3, T-H4, and T-H5), T. hamatum (T-A12), T. koningiopsis (T-K8 and T-K11), and P. cinnamomi (CPO-PCU). In vitro dual-culture assay, the percentage of inhibition of radial growth (PIRG) between Trichoderma spp. and P. cinnamomi strains was measured according to the Bell’s scale. PIRG results indicated that T-AS2 reached the highest value of 78.32%, and T-H5 reached the lowest value of 38.66%. In the greenhouse, the infection was evaluated according to the percentage of disease incidence. Plants with the lowest incidence of dead by avocado root rot were those whose seedlings were inoculated with T-AS2 and T-AS7, resulting in only 5% death by root rot caused by P. cinnamomi. The disease incidence of seedlings with wilt symptoms and death decreased more than 50% in the presence of Trichoderma spp. Relying on the results, we conclude that T. asperellum and T. harzianum contribute to the biocontrol of soil-borne pathogenic oomycete P. cinnamomi.

scholarly journals OPTIMIZING THE PRODUCTION OF A FUNCTIONAL TYPE A RECOMBINANT ENDOCHITINASE FROM Trichoderma asperellum IN Escherichia coli

2021 ◽  
Vol 9 (6) ◽  
pp. 871-880
Author(s):  
Nguyen Ngoc Luong ◽  
Nguyen Quang Duc Tien ◽  
Phung Thi Bich Hoa ◽  
Nguyen Hoang Tue ◽  
Mai Thi Thu Hien ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Soluble Protein ◽  
Total Soluble Protein ◽  
Scale Production ◽  
Trichoderma Asperellum ◽  
Freezing And Thawing ◽  
Trichoderma Spp ◽  
Iptg Induction ◽  
Preparative Scale

Chitinases from the genus Trichoderma fungi are mainly responsible for their anti-fungal activities, which allow them to become the most widely used fungal biocontrol. Therefore, several Trichoderma chitinases have been cloned and expressed to facilitate their production and applications. A previous study of the same authors has characterized an endochitinase from a relatively novel Trichoderma spp., Trichoderma asperellum. To produce this enzyme more economically and efficiently, we reported the synthesis and expression of its synthetic encoding gene in the Escherichia coli M15 strain and established the optimal conditions for preparative scale production of the enzyme in its functional form. By lowering the induction temperatures, we observed substantial improvement in the expression levels of the active enzyme.  At 30 oC and 0.5 mM IPTG induction, 1 L of cells yielded approximately 80 - 100 mg of soluble protein, accounting for about 9-11 % of total soluble protein. This figure may be an underestimation of the actual yield, as deduced from the SDS-PAGE data. The recombinant enzyme can be retrieved by simple repeated freezing and thawing cycles and purified to near homogeneity using Ni-NTA chromatography. The purified enzyme showed in vitro colloidal chitin hydrolysis activity. These results could be scaled up to produce soluble 42 kDa chitinase in E. coli. The study demonstrated an economical method to produce chitinases for various agricultural and environmental applications.

scholarly journals AVALIAÇÃO IN VITRO DA INIBIAÇÃO DO CRESCIMENTO MICELIAL DO FITOPATÓGENO SCLEROTINIA SCLEROTIORUM POR ISOLADOS DE TRICHODERMA SPP.

2021 ◽  
Author(s):  
Kamila Lourrane Carvalho Alencar Rocha ◽  
Kárita Cristine Rodrigues Dos Santos ◽  
Maraiza Castro Bezerra ◽  
Vanice Conceição Do Nascimento
Keyword(s):  
Sclerotinia Sclerotiorum ◽  
Trichoderma Harzianum ◽  
Trichoderma Asperellum ◽  
Trichoderma Spp

Introdução: O fitopatógeno Sclerotinia sclerotiorum, causador da doença conhecida popularmente como mofo-branco, acomete diversas culturas agrícolas em todo o mundo, sendo a soja uma das principais monoculturas mais afetadas. Seu alto grau de patogenicidade está associado à sua capacidade de formar escleródios que atuam como mecanismo de resistência e sobrevivência, podendo permanecer viáveis no solo por mais de dez anos e, em muitos casos, são resistentes aos agroquímicos. Considerando a necessidade de métodos biotecnológicos que contribuam no desenvolvimento agrícola sem promover danos ao ambiente e a população, os agentes de controle biológico (ACBs) são hoje vistos como um excelente artifício. Fungos do gênero Trichoderma podem ser considerados ACBs sagazes por apresentar inúmeros mecanismos com alto potencial antagônico a diversos fitopatógenos, que prejudicam a germinação de esporos, o crescimento das hifas e o desenvolvimento de escleródios. Objetivo: Avaliar o potencial antagônico de isolados de Trichoderma em inibir o crescimento micelial do fitopatógeno S. sclerotiorum através da liberação de metabólitos voláteis. Material e Métodos: Para análise da produção de metabólitos voláteis, os isolados Trichoderma harzianum ALL-42 e o Trichoderma asperellum TR-356 e o fitopatógeno S. sclerotiorum, foram dispostos em fundos de placas de Petri com meio Batata Dextrose Ágar (BDA), em face opostas ao outro, onde, no inferior estava o isolado de Trichoderma e no superior o fitopatógeno. Uma placa controle contendo apenas o fitopatógeno foi feita para comparação dos resultados. Resultados: Ambos isolados de Trichoderma apresentaram resultados positivos na inibição do crescimento micelial de S. sclerotiorum. Conclusão: A produção de metabólitos voláteis é uma ação secundária do gênero Trichoderma, ou seja, sua sobrevivência não depende desse mecanismo, ainda assim, esses metabólitos agem contra os antagonistas presentes no meio. Com isso, podemos verificar o quão eficiente é esse gênero como agente de controle biológico, entretanto, outros isolados precisam ser testados no intuito de encontrarmos agentes mais especialistas na defesa contra o fitopatógeno S. sclerotiorum, e dessa forma instigar os benefícios do emprego de agentes biotecnológicos no campo, visando a redução da aplicação de produtos químicos na agricultura.

scholarly journals Control orgánico in vitro de Phytophthora cinnamomi con aceites esenciales de orégano y clavo

2019 ◽  
Vol 10 (4) ◽  
pp. 961-968
Author(s):  
Yisa María Ochoa Fuentes ◽  
Anselmo Hernández Pérez ◽  
Juan Carlos Delgado Ortiz ◽  
Ernesto Cerna Chavez ◽  
Luis Alberto Aguirre Uribe ◽  
...  
Keyword(s):  
Phytophthora Cinnamomi ◽  
Persea Americana ◽  
San Juan ◽  
Syzygium Aromaticum

  Michoacán es el principal estado productor de aguacate en el mundo; sin embargo, las enfermedades radiculares diezman y dañan los árboles ocasionando su muerte. El objetivo de la presente investigación fue evaluar el control orgánico del crecimiento in vitro de Phytophthora cinnamomi con aceites esenciales de orégano (Lippia berlandieri) y clavo (Syzygium aromaticum). En los meses de octubre y noviembre de 2016, se recolectaron muestras de raíces en árboles con síntomas de la enfermedad en aguacate (Persea americana Mill. var. Hass), en la huerta experimental del INIFAP ubicada en San Juan Nuevo Parangaricutiro, Michoacán. Los aislados se identificaron morfológica y molecularmente. Se evaluó el control de P. cinnamomi con aceites esenciales de orégano y clavo determinando la concentración media inhibitoria y sus límites fiduciales al 95% mediante una regresión Probit por el método de máximas verosimilitud. Los análisis se realizaron utilizando el programa estadístico R 3.4. De acuerdo con los resultados obtenidos, en relación con la inhibición del crecimiento hay una reducción en el crecimiento de P. cinnamomi. Los aceites esenciales de clavo (Syzygium aromaticum) y orégano (Lippia berlandieri) son una alternativa natural para el control del oomiceto P. cinnamomi por su actividad fungicida a bajas concentraciones y pueden incluirse en programas de manejo integrado de enfermedades.

scholarly journals In Vitro and in Planta Evaluation of Trichoderma asperellum TA as a Biocontrol Agent Against Phellinus noxius, the Cause of Brown Root Rot Disease of Trees

Plant Disease ◽  
2019 ◽  
Vol 103 (11) ◽  
pp. 2733-2741 ◽  
Author(s):  
Hao Chou ◽  
Yi-Ting Xiao ◽  
Jyh-Nong Tsai ◽  
Ting-Ting Li ◽  
Hung-Yi Wu ◽  
...  
Keyword(s):  
Root Rot ◽  
White Rot ◽  
Dual Culture ◽  
Trichoderma Asperellum ◽  
In Planta ◽  
Epidemic Disease ◽  
Trichoderma Spp ◽  
Brown Root Rot ◽  
Phellinus Noxius

Brown root rot (BRR), caused by the white rot fungus Phellinus noxius, is an epidemic disease of diverse broadleaved and coniferous tree species in many tropical and subtropical regions. Flooding and trenching control measures are difficult to implement, and chemical controls can have an adverse impact on ecosystems. Previous studies have provided in vitro evidence for the potential use of Trichoderma spp. for biocontrol of BRR. Here, we analyzed the in vitro antagonistic and mycoparasitic abilities of four Trichoderma spp. isolates against four P. noxius isolates in dual culture and Ficus microcarpa wood blocks. A convenient inoculation system based on root inoculation of a highly susceptible loquat (Eriobotrya japonica) with P. noxius-colonized wheat-oat grains was developed to examine the effect of Trichoderma treatment in planta. Preventive application of Trichoderma asperellum TA, the isolate showing high antagonistic activity in vitro, was effective in preventing and delaying the wilting of P. noxius-inoculated loquat cuttings in greenhouse trials. To understand the specific niche in which T. asperellum TA interacts with P. noxius, KOH-aniline blue fluorescence microscopy was used to investigate the colonization of loquat roots by P. noxius and/or T. asperellum TA. Dilution plating assays were also conducted to quantify Trichoderma populations in the rhizosphere and potting mix. T. asperellum TA was able to robustly establish in the rhizosphere and potting mix but with scarce root penetration limited to the superficial layer. We discuss the timing and strategy for applying antagonistic Trichodema sp. on living trees or in BRR-infested areas for BRR management.

Isolation and identification of Trichoderma asperellum, the novel causal agent of green mold disease in sweetpotato

Plant Disease ◽  
2020 ◽  
Author(s):  
Yi-Ling Yang ◽  
BoPing Fang ◽  
Shujie Feng ◽  
Zhang-Ying Wang ◽  
Zhongxia Luo ◽  
...  
Keyword(s):  
Postharvest Disease ◽  
Limiting Factor ◽  
Trichoderma Asperellum ◽  
Storage Roots ◽  
Isolation And Identification ◽  
Trichoderma Spp ◽  
Crop Species ◽  
Molecular Features ◽  
Green Mold

Postharvest disease is an important limiting factor for sweetpotato production. Recently, a new green mold disease was found in sweetpotato storage roots. To investigate the mechanism underlying the pathogenesis of the disease, the pathogen was isolated and identified based on morphological and molecular features, and its characteristics were further analyzed by pathogenic and antagonistic evaluations. The results showed that the isolated pathogen (CRI-Ta1) was identified as Trichoderma asperellum based on the similar growth and morphological features with Trichoderma spp., 99% homology of internal transcribed spacer (ITS) sequence and membership to the same phylogenetic group with the model strain of Trichoderma asperellum (CBS 433.97). The pathogenic analysis revealed that CRI-Ta1 could cause green mold disease through wound infection on the storage roots and the strains re-isolated from infected storage roots could cause disease in different sweetpotato varieties, which was fulfilled in Koch’s postulate. Moreover, CRI-Ta1 could also infect other common crop species, including chestnut, carrot, apple, pear, and others. It indicated that CRI-Ta1 was the pathogen to the storage roots of sweetpotato and had a wide host-range. Additionally, in vitro antagonistic evaluation showed that CRI-Ta1 effectively inhibited the growth of common sweetpotato pathogens, including Fusarium solani and Rhizopus nigricans. However, further research is needed on the potential of CRI-Ta1 to control sweetpotato diseases in vivo. Collectively, our findings provided valuable insights into the characteristics of the Trichoderma asperellum CRI-Ta1 in sweetpoatao and would be helpful to the prevention and control of sweetpotato green mold disease.

scholarly journals Dinámica del crecimiento in vitro de Phytophthora cinnamomi en medios de cultivo alternativos

2019 ◽  
pp. 331-338
Author(s):  
Anselmo Hernández Pérez ◽  
Yisa María Ochoa Fuentes ◽  
Ernesto Cerna Chávez ◽  
Juan Carlos Delgado Ortiz ◽  
Mariana Beltrán Beach ◽  
...  
Keyword(s):  
Phytophthora Cinnamomi ◽  
Persea Americana ◽  
San Juan

México es el principal país productor, consumidor y exportador de aguacate el en mundo. El estudio de la macrobiótica del suelo cobra mayor importancia en el complejo marchitez del aguacate debido que en esta patología pueden estar implicados agentes causales de tipo biótico y abiótico, siendo el Oomycete Phytophthora cinnamomi Rands la principal enfermedad radical del aguacate. El objetivo de la presente investigación fue evaluar la dinámica del crecimiento in vitro de Phytophthora cinnamomi en medios de cultivo alternativos. En los meses de octubre y noviembre de 2016, se recolectaron muestras de raíces en arboles de aguacate (Persea americana Mill. var. Hass) en la huerta experimental del INIFAP ubicada en San Juan Nuevo Parangaricutiro, Michoacán. Los aislados se identificaron morfológica y molecularmente. Se registró las medidas de diámetros de crecimiento de cada cepa in vitro en función al tiempo. Se efectúo una regresión lineal mediante modelos lineales generalizados (GLM) para obtener los parámetros de intercepto y pendiente, significancia de la regresión y el valor de R2 ajustada. Los análisis se realizaron con el programa estadístico R 3.4. De acuerdo con los resultados obtenidos, el medio de cultivo Centeno-agar fue el más eficiente para la proliferación de P. cinnamomi.

scholarly journals ANTAGONISMO DE Trichoderma spp. EN HONGOS ASOCIADOS AL DAÑO DE Diatraea saccharalis Fabricius. (LEPIDOPTERA : CRAMBIDAE ) EN MAIZ

2019 ◽  
Vol 34 (2) ◽  
pp. 17
Author(s):  
JOSE LUIS ARISPE VAZQUEZ ◽  
ABIEL SANCHEZ ARIZPE ◽  
MA. ELIZABETH GALINDO CEPEDA ◽  
MARIO ERNESTO VAZQUEZ BADILLO ◽  
ARNOLDO OYERVIDES GARCIA ◽  
...  
Keyword(s):  
Trichoderma Harzianum ◽  
Trichoderma Asperellum ◽  
Trichoderma Spp ◽  
Fusarium Equiseti ◽  
Trichoderma Longibrachiatum ◽  
Diatraea Saccharalis ◽  
Alternaria Arborescens

El objetivo del presente trabajo, fue evaluar in vitro, mediante cultivo dual la capacidad antagónica de las cepas de Trichoderma asperellum T11, Trichoderma harzianum T1 4, y Trichoderma longibrachiatum T1 40 sobre hongos asociados; Alternaria arborescens, Bipolaris shoemakeri, Bipolaris victoriae, Epicocum sorghinum, Exserohilum longirostratum, Fusarium brevicatenulatum, Penicillium polonicum, Phaeocytostroma ambiguum y Fusarium equiseti, el muestreo se realizó en Tepalcingo, Morelos en tallos de maíz y el experimento se estableció en el laboratorio de Fitopatología de la Universidad Autónoma Agraria Antonio Narro en el mes de abril de 2018. La evaluación se realizó bajo un diseño factorial AxB, con nueve niveles en A y tres en B con cuatro repeticiones por tratamiento, siendo A las cepas de hongos fitopatógenos y B las tres cepas de Trichoderma, se colocó en el extremo de la placa de Petri un explante de PDA con micelio de Trichoderma spp. de 5 mm de diámetro, y en el extremo opuesto un explante del hongo fitopatógeno, las siembras fueron incubadas a 25 ± 2 ºC por 120 h, evaluándose las medias del Porcentaje de Inhibición en el Crecimiento del Micelio (PICM) mediante la fórmula de Fakhrunnisa modificada y se determinó el área de desarrollo de los hongos mediante el software GeoGebra Classic versión 5.0.473.0-d, los resultados se analizaron con el programa (FAUANL) versión 2.5, mediante Tukey con nivel de significancia de 0.05, con efectos de antagonismo mayores del 67.74%.

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