A standard method for the assessment of cell viability has been developed for marine phytoplankton using an inexpensive stain, fluorescein diacetate (FDA), at .75 microM for 10 min. A flow cytometer was used as the fluorescence detector, providing an assessment of viability for each individual particle. Cell size and chlorophyll fluorescence per cell were assessed simultaneously, permitting an assignment of viability to specific subpopulations, thus increasing the power of the technique. A reasonable correspondence between FDA mean fluorescence intensity per cell and an independent metabolic indicator, photosynthetic capacity measured by 14C, was found. Both FDA mean fluorescence intensity and photosynthetic capacity vary as a function of cell volume. Recovery after extended periods of darkness indicate that cells that are FDA negative may not be dead, but merely quiescent or inactive.

Many Trichoderma species are well-known for their ability to promote plant growth and defense. We study how the interaction of bean plants with R. solani and/or Trichoderma affect the plants growth and the level of expression of defense-related genes. Trichoderma isolates were evaluated in vitro for their potential to antagonize R. solani. Bioassays were performed in climatic chambers and development of the plants was evaluated. The effect of Trichoderma treatment and/or R. solani infection on the expression of bean defense-related genes was analyzed by real-time PCR and the production of ergosterol and squalene was quantified. In vitro growth inhibition of R. solani was between 86 and 58%. In in vivo assays, the bean plants treated with Trichoderma harzianum T019 always had an increased size respect to control and the plants treated with this isolate did not decrease their size in presence of R. solani. The interaction of plants with R. solani and/or Trichoderma affects the level of expression of seven defense-related genes. Squalene and ergosterol production differences were found among the Trichoderma isolates, T019 showing the highest values for both compounds. T. harzianum T019 shows a positive effect on the level of resistance of bean plants to R. solani. This strain induces the expression of plant defense-related genes and produces a higher level of ergosterol, indicating its ability to grow at a higher rate in the soil, which would explain its positive effects on plant growth and defense in the presence of the pathogen.

Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents (BCAs) that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defense response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defense-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from the 48 bean genes initially analyzed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34) or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected. As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defense, which respond to the presence of a BCA or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defense genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani.

Xylotrechus arvicola is an important pest in vineyards (Vitis vinifera) in the main Iberian wine-producing regions, and Acanthoscelides obtectus causes severe post-harvest losses in the common bean (Phaseolus vulgaris). Under laboratory conditions with a spray tower, the susceptibility of the immature stages of X. arvicola and A. obtectus against the entomopathogenic fungi Beauveria bassiana and four strains of Trichoderma spp. was evaluated. Both insect pests T. harzianum and B. bassiana showed a good inhibitory activity, accumulating an inhibition on the eggs of values above 85 and 82%, respectively. T. atroviride and T. citrinoviride had a lower inhibitory activity, with inhibition values of 74.1 and 73.3% respectively. These fungi can be considered a highly effective tool for the control during the immature stages of these species.

Xylotrechus arvicola(Olivier) is an emerging pest in vineyards (Vitis vinifera) in the Iberian Peninsula. Its reproductive biology has been studied in the laboratory in order to develop pest-control measures. Obtained results show that the oviposition period is significantly longer in wild females (16.6 ± 2.9 d) than in laboratory females (9.7 ± 1.3 d), but there are no significant differences in the egg-hatching periods between the wild and the laboratory females. Fecundity is significantly higher in laboratory females (93.9 ± 12.1 eggs per female) than in wild females (56.8 ± 9.2 eggs per female). On the other hand, viability (percentage of viable eggs per female) is significantly higher in wild females (53.7 ± 6.6%) than in laboratory females (22.9 ± 3.6%). Wild females lay fewer eggs over a longer period, while laboratory females concentrate on laying in the first 6 d.X. arvicolafemales (wild and laboratory) lay the highest number of eggs on the sixth day. The fecundity and viability of eggs are extended over a longer period in wild females than in laboratory females. SoX. arvicolais a species in which larvae feed directly affects reproductive patterns. These results suggest that, when larvae feed directly from the natural host, the fecundity period and the viability of eggs increase. So the selection of a host plant could be influenced by the composition of the wood. This behavior of the species can also help to advance knowledge of the biology and ecology of this pest, to be applied in integrated control.

<p>The Spanish National Plant Genetic Resource Center’s core collection of bean germplasm includes 202 accessions selected from more than 3000 accessions in function of passport data, seed phenotype, genetic background, and agronomic traits. To acquire more useful information about these accessions, we cultivated and characterized them for sensory and culinary traits. We found considerable variation for culinary and sensory traits of the cooked beans (mean coefficients of variation: 41% for the sensory traits and 40% for the culinary traits). The large dataset enabled us to study correlations between sensory and culinary traits and among these traits and geographic origin, seed color, and growth habit. Greater proportion of white in the seed coat correlated positively with brightness and negatively with mealiness (r=0.60, r=-0.60, <em>p</em>&lt;0.001, respectively). Mealiness correlated negatively with seed-coat roughness and rate of water absorption (r=-0.60, r=-0.53, <em>p</em>&lt;0.001, respectively). Materials of Andean origin had lower seed-coat brightness (<em>p</em>&lt;0.01) and seed-coat roughness, and greater seed-coat perceptibility, mealiness, flavor, and aroma (<em>p</em>&lt;0.001) than materials of Mesoamerican origin. Growth habit failed to correlate with culinary or sensory traits. Breeders can benefit from the information about this core collection available at <a href="http://wwwsp.inia.es/Investigacion/centros/crf/BasesDatos/Bases">www.crf.inia.es/crfesp/paginaprincipaljudia</a>.asp.</p>

Background and Aims An important factor for the success of a mass capture strategy to control cerambycids involves the selection of an effective trap–lure combination. Therefore, the aim was the evaluation of traps with lures and their efficacy for monitoring and mass trapping of Xylotrechus arvicola. Methods and Results Three trap types, Crosstrap, Delta and Screen‐adhesive, with lures baited with ethanol, were evaluated during 4 years in vineyards of two cultivars. The Crosstrap obtained the greatest catches during 2013 in Tempranillo (29.6 adults/trap) and Prieto Picudo (12.9 adults/trap). All trap types showed similar periods of greatest catches in both cultivars. Conclusions The Crosstrap achieved the greatest number of adults captured in both cultivars. The Delta and Screen‐adhesive had few catches. The Tempranillo has proved to be more susceptible to attack by this insect than Prieto Picudo. The capture period varies from 30 to 50 days in the same cultivar in different years. Low temperature during spring delays the appearance of adults. Significance of the Study The Crosstrap baited with ethanol captured the greatest number of X. arvicola adults. This trap can be used to improve monitoring of adult emergence and for controlling this pest by mass trapping.

Xylotrechus arvicola (Olivier) (Coleoptera: Cerambycidae) is an important pest in vineyards (Vitis vinifera) in the main Iberian wine‐producing regions. Larvae were reared with Semi‐Synthetic Iglesias (SSI) diet over 27 months and two generations in the laboratory. Larval mortality was highest during the first (49.49 %) and second (9.38 %) month of rearing, increasing to 50.52 % during the first month if F2 reared larvae were obtained from an F1 adult female obtained in laboratory. The diet had sufficient nutrients to enable the pest to complete its life cycle within nine months, with F1 larval viability ranging from 23.49 % to 27.97 % and F2 larval viability reduced to 2.07 %. However, the diet did not allow for the completion of additional life cycles and generations (F3, F4,…). Larval mortality increased as the months of rearing (66.13 %, 69.51 % and 89.50 %) and generations (59.10 % and 76.93 % in F1 and F2, respectively) progressed in the laboratory. The larva–adult period of females obtained in the laboratory was longer than for males. In the laboratory, the life cycle was shortened in relation to the life cycle in the field because larvae did not require a cold period to break diapause and start pupation. This indicates that X. arvicola has the potential to complete its life cycle inside grape wood in vineyards of wine‐producing regions with warmer winters.