The complete genomic sequence of a variant of the recently reported maize-associated totivirus (MATV) from China was obtained from commercial maize in Ecuador. The genome of MATV-Ec (Ecuador) (4,998 bp) is considerably longer than that of MATV-Ch (China) (3,956 bp), the main difference due to a ≈ 1-kb-long capsid-protein-encoding fragment that is completely absent from the Chinese genome. Sequence alignments between MATV-Ec and MATV-Ch showed an overall identity of 82% at the nucleotide level, whereas at the amino acid level, the viruses exhibited 95% and 94% identity for the putative capsid protein and the RNA-dependent RNA polymerase (RdRp), respectively. Phylogenetic analysis of the viral RdRp domain indicated that MATV-Ec and MATV-Ch share a common ancestor with other plant-associated totiviruses, with Panax notoginseng virus A as the closest relative. MATV-Ec was detected in 46% (n = 80) of maize plants tested in this study, but not in endophytic fungi isolated from plants positive for the virus.
The root-knot nematode, Meloidogyne spp., is an endoparasite that infects plants' root system and causes yield losses in several important crops. Meloidogyne is one of the most devastating pests, so searching for effective biological agents is needed to mitigate its damage. In this study, the predatory activity of Arthrobotrys oligosporus Fresen strain C-2197, obtained from a tropical dry forest of Ecuador, was evaluated as a biocontrol alternative for root-knot caused by Meloidogyne spp. Our results showed that A. oligosporus C-2197 has predatory activity against juvenile nematodes, 72.31%, and 79% efficacy, for in vitro and greenhouse conditions. Besides, the studied strain showed growth-promoting activity, increasing leaf and root area of inoculated plants. Growth promoting activity was also observed in field tests. The present study validates the potential use of A. oligosporus as a biocontrol of Meloidogyne spp. in tomato production systems under greenhouse. It also presents useful information on the use of different cultivation media and substrates for massive A. oligosporus spore concentrates.
One of the main problems in the production of cacao in Ecuador is the disease caused by the fungus Moniliophthora roreri (frosty pod rot) which affects the pods. Here, we evaluate the genetic diversity of this fungus in Ecuador, and its sensitivity to the fungicide azoxystrobin. We evaluated 76 monosporic cultures from the Amazon and the Pacific coast regions. In vitro sensitivity assays tested several doses of the fungicide azoxystrobin to determine the percent of growth inhibition and the IC50. Concentrations of 1 to 0.1 µg mL−1 inhibited the growth of at least 91% of the isolates. Three isolates were the less sensitive (IC50 = 0.0220–0.0364 µg mL−1), two from Guayas (Pacific coast) and one from Sucumbíos (Amazon) provinces. However, M. roreri is highly sensitive, and it could be used in integrated management of the disease. Genetic analyses were carried out by amplifying microsatellite markers (SSR). All the genetic diversity statistics show a higher diversity in the Amazon compared to samples of the coast region; however, the molecular variance was low (FST = 0.11). Discriminant analysis clearly distinguishes three clusters concurrent with the provinces (Sucumbíos, Orellana and El Oro) and a group with the rest of the provinces. Minimum spanning networks shows, unexpectedly, that M. roreri from the coast were derived from at least three independent introductions from the Amazon. Findings are discussed in light of previous Pan-American genetic studies and available historical reports.
La adaptabilidad de los hongos endófitos a sus hospedantes, los beneficios ecológicos que le brinda y los diversos mecanismos antagónicos contra plagas que poseen los convierten en una alternativa para el control biológico de enfermedades. Se determinó el potencial de 17 cepas de hongos endofíticos foliares (FEF) obtenidas de tejido sano de Theobroma cacao como candidatas para el control biológico de Moniliophthora roreri (MR) y M. perniciosa (MP). Se evaluaron: i) el micoparasitismo de los FEF frente a colonias de Moniliophthora spp., ii) la acción de los metabolitos crudos de los FEF en el crecimiento, y iii) la habilidad para recolonizar hojas sanas del hospedante mediante ensayos de hojas sueltas. Tres cepas de Lasiodiplodia theobromae fueron las más promisorias: Ec098, Ec151 and Ec157. Estas cepas inhibieron el crecimiento de MR y MP, tanto en el enfrentamiento de las colonias como mediante sus metabolitos y, adicionalmente, recolonizaron el hospedante entre el 80-100 % de las veces. Otras cepas mostraron valores destacados en un indicador, y no deseables en otros. Por ejemplo, la Ec035 (L. theobromae) mostró los niveles más altos de micoparasitismo contra ambos patógenos en la interacción de las colonias, y el segundo mejor por sus metabolitos, pero no pudo reinfectar el hospedante. La cepa Ec059 (Xylaria feejeensis) reinfectó 100 %, pero no mostró los atributos deseados de antagonismo. Por su parte, los metabolitos de Ec107 (Colletotrichum gloeosporioides s.l.) inhibieron a MR en un 60 %, pero también estimularon el crecimiento de MP. Ninguna cepa logró todas las características deseables para un agente de control biológico.
Frosty pod rot (FPR) of cacao is caused by Moniliophthora roreri (MR). Effective management must include chemical or biological control, in addition to agronomic tactics. Flutolanil has been effective in controlling FPR. The objective of this research was to determine the response to flutolanil of MR isolates from the Amazon and Coast regions of Ecuador. Percentage of mycelial growth inhibition (PGI), and medium inhibitory concentration (IC50) against three concentrations of the fungicide were determined. One µg mL-1 of flutolanil in the culture medium inhibited completely the growth of the 76 MR isolates. At 0.1 μg mL-1, 74/76 were inhibited between 70-97 % compared to the control. Conversely, at the lower concentrations (0.01-0.001 μg mL-1) the results of inhibition were only of 22 % and 47 %, respectively. Four groups were identified: one included the most sensitive (66/76) to different degrees, and three groups representing 10 isolates that were stimulated at low concentrations of flutolanil. IC50 values were low, indicating very high sensitivity in the MR population. IC50max were 0.1342 and 0.1457 in two isolates from the Coast and the Amazon regions, respectively. Isolates from the Coast were significantly less sensitive to flutolanil than those from the Amazon ( IC50 = 0.046 ± 0.03 and IC50 = 0.030 ± 0.02, respectively), however, the differences were minimal. There were no significant differences when comparing the provinces. The most stimulated isolates were found in the provinces of Orellana and Los Ríos. It is concluded that flutolanil seems effective against the causal agent of FPR in Ecuador, both Coast and Amazon regions.
Ecuador is one of the major cocoa producers worldwide, but its productivity has lately been affected by diseases. Endophytic biocontrol agents have been used to minimize pathogenic effects; however, compounds produced by endophytes are minimally understood. This work presents the chemical characterization of the Trichoderma species extracts that proved inhibition against cocoa pathogens. Solid–liquid extraction was performed as a partitioning method using medium with the fungal mycelia of Trichoderma reesei (C2A), Trichoderma sp. (C3A), Trichoderma harzianum (C4A), and Trichoderma spirale (C10) in ethyl acetate individually. The extract of T. spirale (C10) exhibited the growth inhibition (32.97–47.02%) of Moniliophthora perniciosa at 10 µg/mL, while a slight stimulation of Moniliophthora roreri was shown by the extracts of T. reesei (C2A) and T. harzianum (C4A) at higher concentrations. The inhibitory activity could be related to alkaloids, lactones, quinones, flavonoids, triterpenes, and sterols, as indicated by chemical screening and antifungal compounds, such as widdrol, β-caryophyllene, tyrosol, butyl isobutyrate, sorbic acid, palmitic acid, palmitelaidic acid, linoleic acid, and oleic acid, which were identified by gas chromatography–mass spectrometry (GC-MS). The results showed that the extracts, particularly T. spirale (C10), have the potential as biocontrol agents against witches’ broom disease; however, further studies are needed to confirm their effectiveness.
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