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Background Andean lupin (Lupinus mutabilis Sweet) is an important leguminous crop from South America with a high protein content. In Ecuador, lupin yields are severely affected by the infestation of Delia platura larvae on germinating seeds. The application of elicitor molecules with activity against herbivorous insects to control Delia platura infestation constitutes an unexplored and promising alternative for chemical insecticides. In this study, Methyl Jasmonate (MeJA), Hexanoic acid (Hxa), Menadione Sodium Bisulfite (MSB), and DL-β-aminobutyric acid (BABA) were evaluated for their ability to induce resistance against D. platura in three commercial lupin cultivars. Results Only seeds pre-treated with MeJA significantly impaired insect performance during choice and no-choice assays. Additionally, fitness indicators such as seed germination and growth were not affected by MeJA treatment. To investigate the molecular mechanisms behind the MeJA-mediated resistance, RT-qPCR assays were performed. First, RT-qPCR reference genes were validated, showing that LmUBC was the most stable reference gene. Next, expression analysis over time revealed that MeJA application upregulated the activity of the Jasmonic Acid biosynthetic genes LmLOX2 and LmAOS, together with other jasmonate-related defense genes, such as LmTPS1, LmTPS4, LmPI2, LmMBL, LmL/ODC, LmCSD1, and LmPOD. Conclusion This study indicates that MeJA can be used as an environmentally friendly elicitor molecule to protect Andean lupin from D. platura attack without fitness cost. MeJA application induces plant defense responses to insects in Andean lupin that may be modulated by the onset of terpenoid biosynthesis, proteinase inhibitors, lectins, polyamines, and antioxidative enzymes. This article is protected by copyright. All rights reserved.

The plant hormones salicylic acid (SA) and jasmonic acid (JA) regulate defense mechanisms capable of overcoming different plant stress conditions and constitute distinct but interconnected signaling pathways. Interestingly, several other molecules are reported to trigger stress-specific defense responses to biotic and abiotic stresses. In this study, we investigated the effect of 14 elicitors against diverse but pivotal types of abiotic (drought) and biotic (the chewing insect Ascia monuste, the hemibiotrophic bacterium Pseudomonas syringae DC 3000 and the necrotrophic fungus Alternaria alternata) stresses on broccoli and Arabidopsis. Among the main findings, broccoli pre-treated with SA and chitosan showed the highest drought stress recovery in a dose-dependent manner. Several molecules led to increased drought tolerance over a period of three weeks. The enhanced drought tolerance after triggering the SA pathway was associated with stomata control. Moreover, methyl jasmonate (MeJA) reduced A. monuste insect development and plant damage, but unexpectedly, other elicitors increased both parameters. GUS reporter assays indicated expression of the SA-dependent PR1 gene in plants treated with nine elicitors, whereas the JA-dependent LOX2 gene was only expressed upon MeJA treatment. Overall, elicitors capable of tackling drought and biotrophic pathogens mainly triggered the SA pathway, but adversely also induced systemic susceptibility to chewing insects. These findings provide directions for potential future in-depth characterization and utilization of elicitors and induced resistance in plant protection.

Understanding the major drivers that influence soil bacterial and fungal communities is essential to mitigate the impacts of human activity on vulnerable ecosystems, like those found on the Galápagos Islands. Located ~1000 km off the coast of Ecuador, the volcanically formed islands are situated within distinct oceanic currents, which provide seasonal weather patterns and unique microclimates within small spatial scales across the islands. Although much is known about the impacts of human activity, such as climate change and invasive plant species, on above ground biodiversity of the Galápagos Islands, little is known about the resident soil microbial communities and the drivers that shape these communities. Here, our goal was to investigate the bacterial and fungal communities found in soil located in three distinct microclimates: Mirador (arid), Cerro Alto (transition zone), and El Junco (humid), and associated with native and invasive plant types. At each site, we collected soil at three depths (rhizosphere, 5 cm, and 15 cm) associated with the invasive plant, Psidium guajava (guava), and native plant types. We determined that the sampling location (microclimate) was the strongest driver of both bacterial and fungal communities (74 and 38%, respectively), with additional minor but significant impacts from plant type and soil depth. This study highlights the continued need to explore microbial communities across diverse environments and demonstrates the weight of different abiotic and biotic factors impacting soil microbial communities across San Cristóbal Island in the Galápagos archipelago.IMPORTANCE/SIGNIFICANCEHuman activity such as climate change, pollution, introduction of invasive species, and deforestation, poses a huge threat to biodiverse environments. Soil microbiota are an essential component to maintaining healthy ecosystems. However, a greater understanding of factors that alter these microbial communities is needed in order to find ways to mitigate and reverse the impacts imposed by human activity. The Galápagos Islands are a unique real-world laboratory, in that the islands’ biogeography and physical locations in the Pacific Ocean provide distinct microclimates within small geographic distances. Harnessing these distinct environments allowed us to investigate the influence of microclimates, soil depth, and vegetation cover on bacterial and fungal community composition.

Biological treatment using sulfate-reducing bacteria (SRB) is a promising approach to remediate acid rock drainage (ARD). Our purpose was to assess the performance of a sequential system consisting of a limestone bed filter followed by a sulfate-reducing bioreactor treating synthetic ARD for 375 days and to evaluate changes in microbial composition. The treatment system was effective in increasing the pH of the ARD from 2.7 to 7.5 and removed total Cu(II) and Zn(II) concentrations by up to 99.8% and 99.9%, respectively. The presence of sulfate in ARD promoted sulfidogenesis and changed the diversity and structure of the microbial communities. Methansarcina spp. was the most abundant amplicon sequence variant (ASV); however, methane production was not detected. Biodiversity indexes decreased over time with the bioreactor operation, whereas SRB abundance remained stable. Desulfobacteraceae, Desulfocurvus, Desulfobulbaceae and Desulfovibrio became more abundant, while Desulfuromonadales, Desulfotomaculum and Desulfobacca decreased. Geobacter and Syntrophobacter were enriched with bioreactor operation time. At the beginning, ASVs with relative abundance

Anthocyanins are plants metabolites that are recognized by its red/purple coloration produced in flowers, seeds and leaves. These molecules are potentially important to the industry for its antioxidant capacity, disease prevention and as a natural dye. Currently, the production of anthocyanins is carried out using in vitro culture of Vitis vinifera and its yield is increased by using elicitors or stress factors. Zea mays is relevant due to its high content of cyanidin-3-β-glucoside anthocyanin. In the present study the production of cyanidin-3-β-glucoside was evaluated with different mechanisms of elicitation using in vivo and in vitro culture of purple and white maize varieties. The highest callus induction (85%) for white maize was obtained in MS medium supplemented with 2 mg/L of 2,4-Dichlorophenoxyacetic acid, while for purple maize (93%) was obtained in N6 medium with 2 mg/L of 2,4-Dichlorophenoxyacetic acid, using germinated seed as explant for both varieties. Methyl jasmonate was evaluated as an elicitation tool, however no cyanidin-3-β-glucoside was found to be accumulated or produced in vitro. In contrast, using germinated seeds and radicle tissue, elicitation using phosphorus deficiency treatment produced the highest cyanidin-3-β-glucoside accumulation (0.06 mg g–1) in white maize. No elicitation and further production of anthocyanins was found when purple maize were used using this method. Therefore, in vivo elicitation in white maize is a potential method to produce a stable anthocyanin that could be optimized for future applications.

Since the introduction of glyphosate-resistant crops, glyphosate has become the most common and widely used herbicide around the world. Due to its intensive use and ability to bind to soil particles, it can be found at low concentrations in the environment. The effect of these remnants of glyphosate in plants has not been broadly studied; however, glyphosate 1,000 to 100,000 times less concentrated than the recommended field dose promoted growth in several species in laboratory and greenhouse experiments. However, this effect is rarely observed in agricultural fields, where complex communities of microbes have a central role in the way plants respond to external cues. Our study reveals how root-associated bacteria modulate the responses of Arabidopsis to low doses of glyphosate, shifting between growth promotion and growth inhibition., Glyphosate is a commonly used herbicide with a broad action spectrum. However, at sublethal doses, glyphosate can induce plant growth, a phenomenon known as hormesis. Most glyphosate hormesis studies have been performed under microbe-free or reduced-microbial-diversity conditions; only a few were performed in open systems or agricultural fields, which include a higher diversity of soil microorganisms. Here, we investigated how microbes affect the hormesis induced by low doses of glyphosate. To this end, we used Arabidopsis thaliana and a well-characterized synthetic bacterial community of 185 strains (SynCom) that mimics the root-associated microbiome of Arabidopsis. We found that a dose of 3.6 × 10−6 g acid equivalent/liter (low dose of glyphosate, or LDG) produced an ∼14% increase in the shoot dry weight (i.e., hormesis) of uninoculated plants. Unexpectedly, in plants inoculated with the SynCom, LDG reduced shoot dry weight by ∼17%. We found that LDG enriched two Firmicutes and two Burkholderia strains in the roots. These specific strains are known to act as root growth inhibitors (RGI) in monoassociation assays. We tested the link between RGI and shoot dry weight reduction in LDG by assembling a new synthetic community lacking RGI strains. Dropping RGI strains out of the community restored growth induction by LDG. Finally, we showed that individual RGI strains from a few specific phyla were sufficient to switch the response to LDG from growth promotion to growth inhibition. Our results indicate that glyphosate hormesis was completely dependent on the root microbiome composition, specifically on the presence of root growth inhibitor strains. IMPORTANCE Since the introduction of glyphosate-resistant crops, glyphosate has become the most common and widely used herbicide around the world. Due to its intensive use and ability to bind to soil particles, it can be found at low concentrations in the environment. The effect of these remnants of glyphosate in plants has not been broadly studied; however, glyphosate 1,000 to 100,000 times less concentrated than the recommended field dose promoted growth in several species in laboratory and greenhouse experiments. However, this effect is rarely observed in agricultural fields, where complex communities of microbes have a central role in the way plants respond to external cues. Our study reveals how root-associated bacteria modulate the responses of Arabidopsis to low doses of glyphosate, shifting between growth promotion and growth inhibition.

In Ecuador, strawberry production is located in the Andean region with an area of 1000 ha. Albion is the most popular cultivar due to its conical fruit shape, fruit size, bright red color, and swee...

Ancient societies first attempted to domesticate wild plants for food production, which gave rise to present-day agriculture. Nowadays, more than a third of agricultural production is lost due to abiotic or biotic stress, such as drought, salinity, pests and diseases. Current predictions indicating an expanding world population until the end of the century, one of humanity's greatest challenges should be how to feeding the world in a sustainable way, as future increases in crop yields should be achieved with fewer input of fertilizers and pesticides. These challenges have raised awareness of the importance of the plant root microbiome to improve agricultural and horticultural practices. Plant roots are colonized by a surprising number of microorganisms, revealing in most cases an intimate symbiotic relationship which facilitates nutrient uptake and provides the host plant with higher resistance against attackers. In this context, plants can be seen as "superorganisms" that depend on their root microbiome for important functions. But the impact of plant domestication on the functional diversity and beneficial activities of this root microbiome is still largely unknown. Recent studies showed taxonomic differences in the root microbiome between wild relatives and modern cultivars, mainly in root architecture and root exudation. This leads to the hypothesis that present-day cultivars might have lost traits to recruit and activate host-specific beneficial root microbiota. Our research project investigates tomato species and native soils in the Andes to explore the taxonomic and functional diversity of their root microbiomes. Next generation sequencing and ‘omics technologies, combined with classic microbiological techniques are being used to obtain insight in the diversity of root-associated microbial communities of tomatoes. We hypothesize that wild tomatoes grown in their native soils harbour unique and higher frequencies of beneficial root microbiota, compared to modern cultivars.

El objetivo del presente estudio fue construir y operar una cámara anaeróbica no costosa para el cultivo de bacterias sulfato reductoras (BSRs) en muestras ambientales de lodos. Una vez construida la cámara, las condiciones anaeróbicas al interior de la cámara fueron evaluadas a través del monitoreo de la concentración de oxígeno y la temperatura. En la cámara anaeróbica se alcanzaron las condiciones de anaerobiosis (< 0,1% O2) y la temperatura requerida (30ºC) para el cultivo de las BSRs. A continuación, se cultivó en placa (en un medio selectivo) cuatro muestras diferentes de lodo que fueron incubadas, contadas y microscópicamente observadas. Los resultados por cultivo en placa fueron comparados con los de contaje directo en cámara de Neubauer. Habiéndose demostrado que las condiciones ambientales en la cámara son las adecuadas para el cultivo de microorganismos anaeróbicos estrictos, las BSRs también fueron observadas mediante microscopía. En base a los resultados obtenidos por cultivo en placa y por recuento en cámara de Neubauer se determinó que no hubo diferencias significativas en la muestra de suspensión bacteriana, a diferencia de las muestras de lodos. Si bien es cierto que el cultivo de microorganismos demanda tiempo, esta técnica sigue siendo útil y confiable para el conteo microbiano. Finalmente, la cámara anaeróbica construida permite el cultivo de microorganismos anaeróbicos estrictos como las BSRs, responsables de la bioprecipitación de metales.

Botrytis cinerea is a fungal pathogen that limits rose production and commercialization worldwide. Therefore, we evaluated a novel postharvest treatment against Botrytis cinerea in roses (Rosa sp. cv Vendela) using coating bases and antifungal agents of natural origin. Aloe vera pulp, cassava starch and gelatin were used as coating bases. Oregano essential oil (Origanum vulgare), thyme essential oil (Thymus vulgaris) and chitosan were used as natural antifungal agents. The coating bases were evaluated in different concentrations to observe effects of toxicity and opening diameter in rose buds. Gelatin and cassava starch coatings inhibited rose opening and showed petal damage in all concentrations tested. However, Aloe vera pulp at 25% allowed normal buds’ opening and no damage was observed, indicating that Aloe vera could be an ideal coating base for rose postharvest treatments. During in vitro assays, natural antifungal agents efficiently inhibited Botrytis cinerea growth in the concentrations tested. Further, mixture treatments of Aloe vera pulp (25%) with oregano essential oil (1%), thyme essential oil (0.1%) and chitosan (0.1%) showed independently neither damage nor opening inhibition in rose buds. Selected combinations of Aloe vera pulp and natural antifungal agents were applied in roses infected with Botrytis cinerea to evaluate their control of this pathogen. Unfortunately, the selected combinations did not reduce pathogen growth during postharvest treatments since they were similar to untreated controls. Further research has to be performed to find ideal combinations with Aloe vera that could inhibit B. cinerea during postharvest treatments in roses.

Memorias del Tercer Simposio en Fitopatología, Control Biológico e Interacciones Planta-Patógeno

Tapeworms Taenia solium and Taenia saginata are the causative agents of taeniasis/cysticercosis. These are diseases with high medical and veterinary importance due to their impact on public health and rural economy in tropical countries. The re-emergence of T. solium as a result of human migration, the economic burden affecting livestock industry, and the large variability of symptoms in several human cysticercosis, encourage studies on genetic diversity, and the identification of these parasites with molecular phylogenetic tools. Samples collected from the Ecuadorian provinces: Loja, Guayas, Manabi, Tungurahua (South), and Imbabura, Pichincha (North) from 2000 to 2012 were performed under Maximum Parsimony analyses and haplotype networks using partial sequences of mitochondrial DNA, cytochrome oxidase subunit I (COI) and NADH subunit I (NDI), from Genbank and own sequences of Taenia solium and Taenia saginata from Ecuador. Both species have shown reciprocal monophyly, which confirms its molecular taxonomic identity. The COI and NDI genes results suggest phylogenetic structure for both parasite species from south and north of Ecuador. In T. solium, both genes gene revealed greater geographic structure, whereas in T. saginata, the variability for both genes was low. In conclusion, COI haplotype networks of T. solium suggest two geographical events in the introduction of this species in Ecuador (African and Asian lineages) and occurring sympatric, probably through the most common routes of maritime trade between the XV-XIX centuries. Moreover, the evidence of two NDI geographical lineages in T. solium from the north (province of Imbabura) and the south (province of Loja) of Ecuador derivate from a common Indian ancestor open new approaches for studies on genetic populations and eco-epidemiology.

Quinoa (Chenopodium quinoa Willd.) is a highly nutritious pseudocereal with an outstanding protein, vitamin, mineral and nutraceutical content. The leaves, flowers and seed coat of quinoa contain triterpenoid saponins, which impart bitterness to the grain and make them unpalatable without postharvest removal of the saponins. In this study, we quantified saponin content in quinoa leaves from Ecuadorian sweet and bitter genotypes and assessed the expression of saponin biosynthetic genes in leaf samples elicited with methyl jasmonate. We found saponin accumulation in leaves after MeJA treatment in both ecotypes tested. As no reference genes were available to perform qPCR in quinoa, we mined publicly available RNA-Seq data for orthologs of 22 genes known to be stably expressed in Arabidopsis thaliana using geNorm, NormFinder and BestKeeper algorithms. The quinoa ortholog of At2g28390 (Monensin Sensitivity 1, MON1) was stably expressed and chosen as a suitable reference gene for qPCR analysis. Candidate saponin biosynthesis genes were screened in the quinoa RNA-Seq data and subsequent functional characterization in yeast led to the identification of CqbAS1, CqCYP716A78 and CqCYP716A79. These genes were found to be induced by MeJA, suggesting this phytohormone might also modulate saponin biosynthesis in quinoa leaves. Knowledge of the saponin biosynthesis and its regulation in quinoa may aid the further development of sweet cultivars that do not require postharvest processing.

Las rosas son las plantas ornamentales de exportación más cultivadas en el Ecuador. En esta investigación llevada a cabo en las zonas de Machachi y La Leticia, provincia de Pichincha, Ecuador, se evaluó el efecto de ambientes diferenciados por el manejo de cortinas tanto en los días a cosecha como en la calidad en rosa de exportación (Rosa sp variedad Freedom) cultivada en invernadero. Se evaluaron dos tratamientos donde se diferenció el manejo de cortinas de los invernaderos: 1. cortinas completamente cerradas, y 2. manejo combinado abertura-cierre. Para determinar los días a cosecha se usaron dos métodos, uno predictivo denominado TEMPSUM (temperatura sumada en °C acumulados requerida para esta cosecha) y otro directo usando los datos reales desde el “pinch”. Los resultados de este análisis indicaron que los datos reales no se correlacionan con los estimados en TEMPSUM. Los parámetros de calidad evaluados fueron largo de tallo, largo y ancho del botón y número de pétalos. Como resultados se observa solo una tendencia en el tratamiento de cortinas completamente cerradas, que favorece la reducción en los días a la cosecha, e incremento de la calidad respecto al largo de tallos en una de las zonas evaluadas. No hubo diferencias significativas en ninguna de las variables evaluadas.

Antagonism between the defense hormones salicylic acid (SA) and jasmonic acid (JA) plays a central role in the modulation of the plant immune signaling network, but the molecular mechanisms underlying this phenomenon are largely unknown. Here, we demonstrate that suppression of the JA pathway by SA functions downstream of the E3 ubiquitin-ligase Skip-Cullin-F-box complex SCFCOI1, which targets JASMONATE ZIM-domain transcriptional repressor proteins (JAZs) for proteasome-mediated degradation. In addition, neither the stability nor the JA-induced degradation of JAZs was affected by SA. In silico promoter analysis of the SA/JA crosstalk transcriptome revealed that the 1-kb promoter regions of JA-responsive genes that are suppressed by SA are significantly enriched in the JA-responsive GCC-box motifs. Using GCC:GUS lines carrying four copies of the GCC-box fused to the β-glucuronidase reporter gene, we showed that the GCC-box motif is sufficient for SA-mediated suppression of JA-responsive gene expression. Using plants overexpressing the GCC-box binding APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factors ERF1 or ORA59, we found that SA strongly reduces the accumulation of ORA59 but not that of ERF1. Collectively, these data indicate that the SA pathway inhibits JA signaling downstream of the SCFCOI1-JAZ complex by targeting GCC-box motifs in JA-responsive promoters via a negative effect on the transcriptional activator ORA59.