ABSTRACT Perennial ryegrass is one of the most important food sources in animal production. However, several pests affect this crop, and one of the primary control strategies is the symbiotic relationships between ryegrass endophyte fungi. This fungus produces alkaloids that exhibit toxic activity against arthropods. Furthermore, the effect of fungi may extend to higher trophic levels, including predators (spiders and/or insects), decreasing their abundance and diversity. Given the importance of spiders and insects as predators, whether the symbiotic interaction between perennial ryegrass and endophyte fungus reduces the abundance and diversity of predators pose an important question. To address this question, natural enemies in perennial ryegrass were collected and analyzed over a year, and the percentage of endophyte fungus was evaluated by the presence of hyphae from two ryegrass cultivars, Jumbo (E-) and Alto AR1 (E+). We observed an 80% endophyte infection rate for (E+) and 0% for (E-). Moreover, 222 individual spiders corresponding to 10 families were identified in both perennial ryegrasses, including 209 individuals for (E-) and 13 for (E+). The most abundant spider family was Lycosidae, representing 71.17% of the total spiders. In addition, 65 insects were collected, corresponding to 6 families, with Carabidae being the most abundant. Furthermore, the Simpson index indicated the dominance of the family Lycosidae. Overall, spider and insect abundance and diversity were reduced in (E+), suggesting a negative effect of the endophyte on predator populations.

Abstract Flavonoid biosynthesis may be affected by plant domestication, with flavonoid production being reduced in proportion to the degree of domestication. In this context, kaempferol (3,4´,5,7-tetrahydroxyflavone) has been identified in the leaves of wild and cultivated Ugni molinae, a berry endemic to Chile. The biosynthetic pathway of kaempferol production begins with naringenin (4´,5,7-trihydroxyflavanone), which is converted to dihydrokaempferol (3,4´,5,7-tetrahydroxyflavanone), catalyzed by flavanone 3ß-hydroxylase (FHT) and then converted to kaempferol by a bifunctional enzyme called flavonol synthase (FLS). Therefore, our study aims to evaluate how FLS activity is affected in murtilla plants that are subjected to the domestication process. Kaempferol was quantified from methanolic extracts of leaf samples collected from both cultivated and wild U. molinae plants using high-performance liquid chromatography, and enzyme extraction was performed to determine FLS activity. The results showed that kaempferol concentration in wild plants from the Soloyo (0.14 μg g-1), Mehuín (0.18 μg g-1) and Queule (0.25 μg g-1) sampling areas was higher than in their cultivated counterparts. Our data are consistent with the FLS activity detected in samples obtained from Manzanal Alto (134.79 pKatal, Soloyo (96.48 pKatal), and Mehuín (119.97 pKatal). These samples also exhibited higher enzymatic activity than their cultivated counterparts. Together, these data suggest that FLS activity is negatively affected by the domestication process.

Resumen La biosíntesis de flavonoides puede verse afectada por la domesticación vegetal, reduciéndose la producción en relación al grado de domesticación. En este contexto, ha sido identificado el kaempferol (3,4´, 5,7-tetrahidroxiflavona) en hojas de Ugni molinae silvestre y cultivada. La ruta biosintética del kaempferol comienza con naringenina (4´, 5,7-trihidroxiflavanona), que se convierte en dihidrokaempferol (3,4´, 5,7-tetrahidroxiflavanona), catalizada por la flavanona ٣ß-hidroxilasa (FHT). Finalmente, se convierte en kaempferol por una enzima bifuncional llamada flavonol sintasa (FLS). Por lo tanto, nuestro estudio tiene como objetivo evaluar cómo se ve afectada la actividad de FLS en plantas de murtilla que están sujetas a un proceso de domesticación. El kaempferol fue cuantificado a partir de los extractos metanólicos de hojas recogidas en plantas cultivadas y silvestres de U. molinae mediante cromatografía líquida de alta resolución. Además, se realizó una extracción de enzimas para determinar la actividad de FLS. Los resultados mostraron que la concentración de kaempferol en las áreas de muestreo silvestre de Soloyo (0.14 μg g-1), Mehuín (0.18 μg g-1) y Queule (0.25 μg g-1) fue mayor que en sus contrapartes cultivadas. Estos datos fueron consistentes con la actividad de FLS determinada en muestras obtenidas de Manzanal Alto (134,79 pKatal), Soloyo (96,48 pKatal) y Mehuín (119,97 pKatal). Estos también exhibieron una mayor actividad enzimática que sus contrapartes cultivadas. Por lo tanto, nuestros datos sugieren que la actividad de FLS se ve reducida por la domesticación vegetal.

The Argentine stem weevil, Listronotus bonariensis, is an important pest in pastures, especially ryegrass (Lolium sp.), compromising grassland persistence and productivity. Both, adult and specially larvae cause severe damage, action that generate the death of the tiller and reducing forage production. Chemical control has limited effectiveness in adult’s insect and negative effects on the environmental and toxicity to pollinators when applied close to the flowering period is observed. Therefore, the main currently strategy is based on the use of cultivars of Lolium endophyte fungus-infected with Neothypodium genus. However, over a decade that there are no reports on the presence of this weevil in southern Chile. Hence, the aim of this research was report the actual incidence of L. bonariensis in ryegrass pastures. The results of two years of prospecting showed that this weevil is currently causing damage to the ryegrass pastures indicating that the effectiveness of species infected with the fungus must be evaluated.

Plant domestication is a process in which plants’ chemical defenses that help them cope with herbivores might decline. Consequences of this process could be reflected in an increase in insect pests. Therefore, we carried out a survey to contrast the diversity, damage indexes and insect assemblages between cultivated and wild "Murtilla" (Ugni molinae) plants. The main scientific question put forward in this paper is as follows: Is there a decrease in diversity and an increase in both insect assemblages and damage indexes associated with the domestication process in U. molinae plants? The objective of this report was to compare the structure of a taxonomic assemblage collected in both wild and cultivated plants and their temporal variation over the year. Seven ecotypes and their respective wild populations were selected for these studies. The results showed higher insect assemblages in wild parents (77.35%) vs. cultivated (22.7%). The damage indexes were also higher in wild plants (0.23). The diversity indexes according to Margalef (12.98), the Shannon index (5.15) and the Simpson index (19.04) were higher in wild plants. Moreover, approximately 60 species were determínate. We detected changes in insect assemblages, damage and diversity indexes that could suggest that murtilla domestication has altered insect assemblages.

M. Chacón-Fuentes, M. Lizama, L. Parra, I Seguel y A. Quiroz. 2016. Diversidad, composición de la comunidad de insectos e índice de daño, en murtillas silvestres y cultivadas. Cien. Inv. Agr. 43(1):57-67. La domesticación vegetal es un proceso en el cual las plantas pueden disminuir su nivel de defensas químicas para combatir herbívoros. Algunas de las posibles consecuencias podrían reflejarse en un incremento en el número y en la diversidad de insectos. Por lo tanto, se realizó un "survey" para comparar el índice de daño y la diversidad de insectos en plantas cultivadas y silvestres de murtilla (Ugni molinae). La principal interrogante de este trabajo fue: ¿La domesticación en plantas de murtilla, disminuirá la diversidad e incrementará el índice de daño y la comunidad de insectos? El objetivo de esta investigación fue comparar la población de insectos asociada a plantas silvestres y cultivadas además de analizar su variación durante el año. Siete ecotipos y sus correspondientes contrapartes silvestres fueron seleccionadas para este estudio. Los resultados mostraron un mayor número de insectos en plantas silvestres (77,35%) vs. (22,7%) cultivadas. El índice de daño fue mayor en plantas silvestres (0,23) comparada con plantas cultivadas. El índice de Margalef mostró la mayor riqueza de insectos en plantas cultivadas. Sin embargo, el índice de Shannon fue mayor (5.15) en plantas silvestres. Además, cerca de 60 especies de insectos fueron determinadas. Se detectaron cambios en la comunidad de insectos, índices de daño y diversidad que podrían sugerir que la domesticación en murtilla ha alterado la comunidad de insectos.

Two trials were established to evaluate the in vitro and ex vitro rooting of selected microshoots of two selected clones of Ugni molinae. In the in vitro experiment, nine treatments were compared, combination of three concentrations of Murashige and Skoog medium (MS) (1, ½ and ¼ strength macronutrients) and three concentration of indole 3-butyric acid (IBA, at 0, 1 and 2 µM). In the ex vitro assay we established for IBA four concentrations (0; 4.9; 9.8 and 19.7 mM). After 30 days total survival, rooting percentage, root length and number of roots per explants were evaluated. Both clones responded differently to the treatments used. Under in vitro conditions, Clone 1 had an average of 97% rooting at ½ MS + 1 µM IBA, whereas Clone 2 obtained a 93% of rooting with ¼ MS + 2 µM IBA. Our results indicate that the decrease in the strength of MS medium and the addition of IBA promotes rooting. With regard to ex vitro conditions, the Clone 1 did not respond to IBA applications, while in Clone 2, IBA promoted the rooting at the concentration of 19.7 mM, with percentages of 90 and 85%, respectively. Considering these results, it is generally recommended to use the ex vitro rooting system because both clones were able to obtain the same high percentage rooting (above 85%) in this system. Ex vitro propagation will also safe costs.

Con el objetivo de evaluar el enraizamiento in vitro y ex vitro de microtallos de dos clones de Ugni molinae, se establecieron dos ensayos. En el experimento de enraizamiento in vitro se contrastaron nueve tratamientos producto de la combinación de tres concentraciones de medio Murashige y Skoog (MS) (1, ½ y ¼ de los macronutrientes diluidos) y tres de ácido 3-indolbutírico (AIB) (0, 1 y 2 µM). En tanto, para el ensayo de enraizamiento ex vitro se contrastaron cuatro tratamientos correspondientes a las concentraciones de 0; 4,9; 9,8 y 19,7 mM de AIB. Después de 30 días se evaluó la supervivencia, enraizamiento, longitud de raíz y número de raíces por explante. El tratamiento que logró el mayor enraizamiento difirió entre ambos clones. En condiciones in vitro el Clon 1 obtuvo un promedio de enraizamiento de 97% con ½ MS + 1 µM de AIB, mientras que el Clon 2 obtuvo un 93% con ¼ de MS + 2 µM de AIB. Nuestros resultados indican que la disminución de sales en el medio de cultivo MS y la adición de AIB favorece el enraizamiento. Con respecto a las condiciones ex vitro, el Clon 1 no respondió a las aplicaciones de AIB, en cambio en el Clon 2, el AIB promovió el enraizamiento a la concentración de 19,7 mM, alcanzando porcentajes de 90 y 85%, respectivamente. En virtud de los resultados obtenidos, se sugiere un enraizamiento ex vitro para los clones 1 y 2, para evitar los costos asociados al enraizamiento in vitro, ya que independiente del sistema utilizado, ambos clones obtuvieron enraizamientos iguales o superiores a un 85% en el mejor de sus tratamientos.

The objective of this study was to stimulate the production of calluses, to observe its typologies and organogenic response under different induction and differentiation mediums in explants from cotyledons, hypocotyls and leaves of Ugni molinae. Higher callus production was obtained with cotyledons in a Murashige and Skoog (MS) medium supplemented with 0.5 mg L-1 of naphthaleneacetic acid (NAA), similar also in explants of hypocotyls with 1.0 and 5.0 mg L-1 ofNAA, and in the control (62, 62, 74 and 64 %, respectively). Larger sizes of calluses were observed in cotyledons and smaller in leaves. Later, calluses were transferred to subcultures without growth regulators, where they were observed green, friable, compact and phenolic, obtaining the higher survival rates in calluses induced with 0.5 mg L-1 of NAA for cotyledon and hypocotyl, and 1.0 mg L-1 of NAA in leaf. Finally, the calluses derived from these treatments were cultivated in a differentiation medium with different concentrations of NAA plus 6-benzylaminopurine (BAP). The best organogenic responses were achieved with cotyledons in a medium without growth regulators and the rhizogenic in leaves with 0.1 mg L-1 of NAA + 1.0 mg L-1 of BAP. These results indicate that it is possible to stimulate different types of calluses and to obtain roots or shoots in U. molinae, which makes it possible to potentially use this technique for the conservation, micro-propagation and plants breeding of an endemic plant species.

El objetivo de este estudio fue estimular la producción de callos, observar su tipología y respuesta organogénica bajo diferentes medios de inducción y de diferenciación en explantes de cotiledón, hipocótilo y hoja de Ugni molinae. Los mayores porcentajes de callos se lograron con cotiledones en un medio Murashige y Skoog (MS) suplementado con 0,5 mg L-1 de ácido naftalenacético (ANA), como también en explantes de hipocótilo con 1,0 y 5,0 mg L-1 de ANA, además del testigo (62, 62, 74 y 64 %, respectivamente). El mayor tamaño de callos se observó en cotiledones y el menor en hojas. Posteriormente, los callos fueron transferidos a un subcultivo sin reguladores de crecimiento, donde se observaron callos verdes, friables, compactos y fenólicos, obteniendo la mayor sobrevivencia en callos inducidos con 0,5 mg L-1 de ANA para cotiledón e hipocótilo y 1,0 mg L-1 de ANA en hoja. Finalmente, los callos derivados de estos tratamientos fueron cultivados en un medio de diferenciación con distintas concentraciones de ANA más 6-bencilaminopurina (BAP). La mayor respuesta caulogénica se logró en callos de cotiledones en un medio sin reguladores de crecimiento y la rizogénica en callos de hojas con 0,1 mg L-1 de ANA + 1,0 mg L-1 de BAP. Estos resultados indican que es posible estimular distintos tipos de callos y obtener raíces o brotes en U. molinae, lo que posibilita su uso potencial para la conservación, micropropagación y mejoramiento vegetal de una especie endémica.