Search

In the last years, our research has focused on the study of brown rot on stone fruit caused by Monilinia spp. in the ‘Valle del Ebro’ (Spain). The epidemiology of this disease was thoroughly investigated in the field and one of the main outcomes was the development of a prediction model that indicates the risk of infection. Furthermore, the epidemiology was also studied in the postharvest phase, providing a lot of information regarding the relevance of the main postharvest handling operations, the identification of fungal population in packing houses, the fruit infection risks and the influence of temperature and humidity on conidia survival. Additionally, many efforts have been orientated to the development of control strategies for both pre and postharvest periods. Traditionally, chemical fungicides have been used to preserve fruit quality over extended periods of storage or transportation. However, the growing public concern over health and environmental hazards associated with high levels of pesticides has resulted in restrictions imposed by legislation and also by distribution companies. In this context, our main goal has been the development of environmentally friendly alternative strategies to synthetic fungicides in order to control brown rot on stone fruit. Although many studies have demonstrated the efficacy of these alternative treatments, only a few of them are currently applied under commercial conditions. Biocontrol agents, natural compounds from different origins and physical means are the main approaches which have been studied, with different success levels. There are several reasons for the limited success of these treatments, such as the inconsistency of results, variability of the efficacy under commercial conditions, low persistence, a narrow spectrum range of activity, the difficulties in developing a shelf-stable formulated product that retains efficacy (in the case of biocontrol agents), and economical and regulatory limitations. Generally, i

[Background] To improve efficacies achieved through biocontrol agent application, there has been increasing interest in studying the efficacy of biocontrol agent with other existing technologies like chemicals or different biocontrol agent combinations. The compatibility of Penicillium frequentans (Pf909) with modern inputs in plant protection like fungicides (chemicals and biologicals) and insecticides is a pre-requisite for developing integrated brown rot management strategies. [Results] The combination of P. frequentans and Bacillus amyloliquefaciens, each with different mechanisms of action, competition and antibiosis, respectively, did not improve the biocontrol of brown rot when each treatment was applied individually. P. frequentans and B. amyloliquefaciens could not be combined together in the same tank, because bacteria inhibited the germination and growth of P. frequentans tubes in suspension. Furthermore, B. amyloliquefaciens also competes with P. frequentans once applied on fruit surfaces. Additionally, bacteria reduced the growth of P. frequentans CFUs on media, at different 25 °C and 30 °C throughout the incubation periods. However, 76% of fungicides and 90% of insecticides commonly used against stone fruit pests are compatible with P. frequentans. [Conclusions] Results indicated that the most commercial chemical pesticides could be simultaneously applied with Pf909 in integrated pest control, but not B. amyloliquefaciens.

[EN] Ampelomyces quisqualis CPA-9 was reported as a promising biocontrol fungus against powdery mildew on cucurbits, although its formulation had not been developed. The present work aimed to develop a fluidised-bed spray-dried formulation of CPA-9 that included film-forming compounds, which could improve the behaviour of the microorganism under practical conditions. Film forming compounds, efficacy, shelf life, physical stability as a function of aw, and solubility in water were determined. A film-forming formulation based on native potato starch and pregelatinised potato starch as carrier and binder was obtained, and sucrose plus skimmed milk were used as protective compounds. Dehydrated conidia maintained the efficacy of fresh cells against Podosphaera xanthii on zucchini leaves, and powdery mildew was significantly reduced compared with control treatment. Despite of results obtained for the glassy state guaranteed the stability of the powder against sticking and caking processes, the shelf life of the product was limited to 4 months of storage at 4 degrees C. On the other hand, the formulation was mainly dispersible, due to the high concentration of native potato starch which reduces the solubility of the product. Results obtained in this work, such as the high efficacy of the developed product, the low cost of the employed substances, the film-forming ability of the rehydrated powder, and finally the effectiveness of the dehydration process, confirmed the potential of the product. Further studies to verify the improvement of CPA-9 behaviour under practical conditions or to improve the storage conditions to enlarge the shelf life of the BCA should be done to consider the developed product as a biocontrol-based product for cucurbits against powdery mildew.

Different treatments based on the biocontrol agent (BCA) Bacillus amyloliquefaciens CPA-8 to control brown rot under field conditions were evaluated as alternative to chemical applications. As part of a well-designed disease program that enables the integration of BCAs into cropping systems, testing of the sensitivity of Monilinia laxa and Monilinia fructicola at different doses of CPA-8 were conducted in stone fruit under laboratory conditions. CPA-8 dose of 107 CFU mL−1 reduced more than 60.0 and 75.5% of brown rot incidence and severity, respectively. Once in the orchard, different degree of biocontrol activity was obtained depending on the inoculum pressure, which was mainly associated with meteorological conditions. Under drastic disease pressure, neither CPA-8 treatment nor the chemicals controlled the disease at harvest and only the chemical treatment reduced postharvest brown rot incidence. However, when Monilinia spp. incidence was close to the standard levels recorded in the area, treatments based on CPA-8 formulations proved to be efficacious. At harvest, BA3, BA4 treatments (CPA-8 optimised products) and PF + BA3 treatment (Penicillium frequentans combined with CPA- 8) reduced Monilinia spp. incidence compared to the control (54.7–64.1%) and similar to the chemicals. At postharvest, almost all CPA-8-based treatments (except PF + BA3) controlled the pathogen with BA4 treatment being as much effective as the chemicals (50.3% of disease reduction). Finally, the population dynamics of CPA-8 on treated fruit surface remained after treatment application, at harvest and at postharvest shelf-life (>104 CFU cm−2). This study highlights the potential of B. amyloliquefaciens CPA-8 as alternative or complementary strategies to control Monilinia spp. © 2017 Elsevier Ltd

[EN] Multiple formulations of known biocontrol agent (BCA) Candida sake, containing different coatingforming polymers and surfactants were tested at different polymer:BCA ratios, in order to improve control of Botrytis cinerea on grapes. The BCA cell viability on the grape surface was analyzed and reduction in disease incidence and severity was determined. Coating-forming solids improved the survival and effi- cacy of C. sake as a BCA against B. cinerea, depending on the polymer type and ratio. The incorporation of surfactants did not improve survival or disease control, although they promoted a better cell dispersion on the grape surface. Cell growth of the antagonist during incubation led to the formation of aggregates, even when surfactants were present. Sodium caseinate and starch were the most suitable polymers to formulate C. sake preparations to obtain coating-forming systems with this BCA and to increase its survival and efficacy at the minimum economic cost of the ingredients. 2016 Elsevier Inc. All rights reserved

Penicillium digitatum and Penicillium expansum are responsible for green and blue molds in citrus and pome fruits, respectively, which result in major monetary losses worldwide. In order to study their infection process in fruits, we successfully introduced a green fluorescent protein (GFP) encoding gene into wild type P. digitatum and P. expansum isolates, using Agrobacterium tumefaciens-mediated transformation (ATMT), with hygromycin B resistance as the selectable marker. To our knowledge, this is the first report describing the transformation of these two important postharvest pathogens with GFP and the use of transformed strains to study compatible and non-host pathogen interactions. Transformation did not affect the pathogenicity or the ecophysiology of either species compared to their respective wild type strains. The GFP-tagged strains were used for in situ analysis of compatible and non-host pathogen interactions on oranges and apples. Knowledge of the infection process of apples and oranges by these pathogens will facilitate the design of novel strategies to control these postharvest diseases and the use of the GFP-tagged strains will help to determine the response of P. digitatum and P. expansum on/in plant surface and tissues to different postharvest treatments.