Abstract Root-knot nematodes are detrimental to agricultural crops and hard to control once established in the field. The objective of this research was to evaluate the growth, sporulation, and capture in vitro of asexual fungi against Meloidogyne sp. Molecular identification of the asexual fungi included in the study was performed by sequencing the ITS1 region of rDNA. The growth and sporulation of Arthrobotrys sp., Dactylellina sp. and Dactylaria sp. was evaluated in five culture media and two types of substrates (rice husks and ground corn). For the evaluation of attraction and capture of the nematodes, second stage juveniles of Meloidogyne sp. All the evaluated fungal isolates grew and sporulated in the culture media and substrates. The fungi Arthrobotrys sp. (C19-1-1) and Dactylellina sp. (C19-48) showed greater effectiveness in attracting and capturing Meloidogyne sp. The evaluated fungal isolates have the potential to colonize different culture media and substrates. In addition, they can form modified and specialized hyphae that control juvenile Meloidogyne sp. nematodes. Therefore, future studies should focus on evaluating fungal isolates against root-knot nematodes under field conditions.

Resumen Los nematodos formadores de agallas son perjudiciales para los cultivos agrícolas. El objetivo de esta investigación fue evaluar el crecimiento, esporulación y captura de hongos asexuales contra Meloidogyne sp. en condiciones in vitro. La identificación molecular de los hongos asexuales se realizó con PCR utilizando secuencias de la región ITS del ADNr. Se evaluó el crecimiento y esporulación de Arthrobotrys, Dactylellina y Dactylaria en cinco medios de cultivos y dos tipos de substratos (cascarilla de arroz y maíz molido). Para la evaluación de atracción y captura de los nematodos se emplearon juveniles de segunda etapa de Meloidogyne sp. Todos los aislamientos fúngicos evaluados crecieron y esporularon en los medios de cultivos y substratos. Los hongos Arthrobotrys sp. (C19-1-1) y Dactylellina sp. (C19-48) mostraron mayor efectividad en la atracción y captura de Meloidogyne sp. Los aislamientos fúngicos evaluados tienen el potencial de colonizar diferentes medios de cultivos y substratos. Además, pueden formar hifas modificadas y especializadas que controlan nematodos juveniles de Meloidogyne sp. Por lo tanto, los futuros estudios se deberían centrar en evaluar los aislados fúngicos contra los nematodos formadores de agallas en condiciones de campo.

ABSTRACT: Ecuador is responsible for up to 70 % of the world’s production of fine flavor cocoa. Although fungal diseases have reduced cocoa production, eco–friendly alternatives have been implemented to control the spread of phytopathogens. This study aimed to explore the potential of endophytic strains isolated from fine flavor Nacional cocoa trees to synthetize compounds with antimicrobial and surface active properties. It was determined that under in vitro conditions, seven strains (DS03, DS07, DS18, DS23, DS31, DS34 and DS50) exhibited antifungal activity that inhibited the mycelial growth of Moniliophthora roreri and Moniliophthora perniciosa. The strains were identified based on their morphological (pattern forming Bacilli) and genetic characteristics using the 16S SSU rRNA gene sequences. Furthermore, we determined that the endophytic strains contained genes that encoded the lipopeptides Fengycin (fenA, fenB, fenC, fenE), Iturin (ituB, ituC, ituD), and Bacyllomicin D (bamC, bamD). The production of biosurfactant compounds was determined by a reduction in surface tension in the mineral medium. Additionally, the biosurfactant crude extract (BCE) was tested for its inhibitory activity against M. perniciosa and M. roreri. The minimal inhibitory concentration (MIC) was determined to be 0.07 and 0.035 mg mL−1, respectively, in both cases in a fungistatic action mode. The swelling granulation and fragmentation of both phytopathogens’ hyphae were observed using the BCE treatment. These results suggested the biocontrol capacity of the endophytic strains, which under in vitro conditions, have the ability to synthesize antifungal and surface active compounds that might be used as an ecological alternative in cocoa plants to prevent infection by phytopathogens.

RESUMEN Los rizobios han sido estudiados por la simbiosis que establecen con las raíces de las leguminosas. Sin embargo, la colonización y promoción del crecimiento en plantas no leguminosas también ha sido demostrada. Este trabajo tuvo como objetivo la identificación bioquímica y molecular de rizobios rizosféricos presentes en la rizosfera de dos cultivares comerciales de maíz. Se obtuvieron aislamientos cultivables en medio Levadura-Manitol-Agar (YMA) a partir del suelo rizosférico y rizoplano de las plantas. Se determinaron las características culturales del cultivo (tamaño, color, mucosidad, etc.) y morfo-tintoriales (forma celular, respuesta a la tinción y esporulación), así como la respuesta de los aislados a ocho pruebas bioquímicas (producción ácido-base, citrato, oxidasa, catalasa, producción de H2S, ureasa, gelatinasa y ensayo oxidativo-fermentativo) con valor predictivo para la identificación de rizobios. Se determinó el género mediante la secuenciación del gen ADNr-16S. Se obtuvieron 81 aislados totales de los cuales el 30.86% mostró las características culturales, morfológicas y tintoriales esperadas para los rizobios y solamente el 20% de estos correspondió al género Rhizobium.

ABSTRACT Rhizobia have been studied for the symbiosis that they establish with the roots of legumes. However, the colonization and promotion of growth in non-leguminous plants has also been demonstrated. The aim of this work was the biochemical and molecular identification of rhizosphere rhizobia present in the rhizosphere of two commercial maize cultivars. Cultivable isolates were obtained in yeast-mannitol-agar (YMA) medium from rhizospheric soil and the rhizoplane. The cultural (size, color, mucus, etc.), morphological, and staining (cell shape, response to staining and sporulation) characteristics were determined as well as isolate responses to eight biochemical tests (acid-base production, citrate, oxidase, catalase, H2S production, urease, gelatinase and the oxidative-fermentative assay) that are valuable for rhizobia identification. The genus was determined by 16S rDNA gene sequencing. We obtained 81 total isolates of which 30.86% showed the cultural, morphological and staining characteristics expected for rhizobia and only 20% of these corresponded to the genus Rhizobium.