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Outbreak response to quarantine pathogens and pests in the European Union (EU) is regulated by the EU Plant Health Law, but the performance of outbreak management plans in terms of their effectiveness and efficiency has been quantified only to a limited extent. As a case study, the disease dynamics of almond leaf scorch, caused by Xylella fastidiosa , in the affected area of Alicante, Spain, were approximated using an individual-based spatial epidemiological model. The emergence of this outbreak was dated based on phylogenetic studies, and official surveys were used to delimit the current extent of the disease. Different survey strategies and disease control measures were compared to determine their effectiveness and efficiency for outbreak management in relation to a baseline scenario without interventions. One-step and two-step survey approaches were compared with different confidence levels, buffer zone sizes, and eradication radii, including those set by the EU legislation for X. fastidiosa . The effect of disease control interventions was also considered by decreasing the transmission rate in the buffer zone. All outbreak management plans reduced the number of infected trees (effectiveness), but large differences were observed in the number of susceptible trees not eradicated (efficiency). The two-step survey approach, high confidence level, and the reduction in the transmission rate increased the efficiency. Only the outbreak management plans with the two-step survey approach removed infected trees completely, but they required greater survey efforts. Although control measures reduced disease spread, surveillance was the key factor in the effectiveness and efficiency of the outbreak management plans. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license., Competing Interests: The author(s) declare no conflict of interest.

We compared the effects of milk-feeding in 288 Holstein calves (72 per group) which were fed twice (2F) or thrice (3F) daily, with or without the addition of hydrogenated fat-embedded calcium gluconate (G) supplemented in the starter food and in the daily diet up to the age of 9 months, on the calves' metabolism, growth, health, and reproductive efficiency up to first pregnancy. The calves received 6 L of milk replacer (130 g/L) and had ad libitum access to water and textured calf starter with or without gluconate. Gluconate supplementation promoted a "catch-up" in growth in supplemented calves compared to their counterparts that did not receive gluconate. Gluconate appeared to reduce animal metabolic stress during key events, such as weaning and transfer into open-door pens, reducing fructosamine (352.61 vs. 303.06 in 3FG and 3F, respectively; p = 0.028) and urea (3F revealed the highest values compared with the other three groups: 19.06 for 3F vs. 13.9 (2F), 13.7 (2FG), and 14.3 (3FG), respectively, p = 0.002) from weaning onwards. The feeding of dairy calves with milk replacer three rather than two times per day tended to be associated with better health from weaning to 4 months old; parameters such as ultrasound lung score and calf health score improved over time ( p < 0.001). Thrice-daily feeding with milk replacer tended to reduce the number of artificial inseminations per pregnancy in heifers by 0.2 points ( p = 0.092). We confirmed significant correlations between early health and growth parameters and reproductive efficiency and a positive correlation between body weight and average daily weight gain and the thickness of the back fat layer in young heifers ( r = 0.245; p < 0.0001; r = 0.214; p < 0.0001 respectively). Our study was conducted on a commercial farm with reasonably effective animal management, so baseline welfare was likely satisfactory.

Red leaf blotch (RLB) of almond, caused by the ascomycete Polystigma amygdalinum , is a severe foliar disease endemic in the Mediterranean Basin and Middle East. Airborne ascospores of P. amygdalinum were monitored from 2019 to 2021 in two almond orchards in Lleida, Spain, and a Bayesian beta regression was used to model its seasonal dynamics. The selected model incorporated accumulated degree-days (ADD), ADD considering both vapor pressure deficit and rainfall as fixed effects, and a random effect for the year and location. The performance of the model was evaluated in 2022 to optimize RLB fungicide programs by comparing the use of model predictions and action thresholds with the standard program. Two variants were additionally considered in each program to set the frequency between applications, based on (i) a fixed frequency of 21 days or (ii) specific meteorological criteria (spraying within 7 days after rainfalls greater than 10 mm, with daily mean temperatures between 10 and 20°C, and with a minimum frequency of 21 days between applications). Programs were evaluated in terms of RLB incidence and number of applications. The program based on the model with periodic fungicide applications was similarly effective as the standard program, resulting only in a 2.6% higher RLB incidence but with fewer applications (three to four, compared with seven in the standard program). When setting the frequency between applications by using the meteorological criteria, a higher reduction in the number of applications (two to three) was observed, while RLB incidence increased by roughly 16% in both programs. Therefore, the model developed in this study may represent a valuable tool toward a more sustainable fungicide schedule for the control of almond RLB in northeast Spain., Competing Interests: The author(s) declare no conflict of interest.

Huanglongbing (HLB) is one of the most devastating citrus diseases worldwide. It is associated with the non-culture bacteria Candidatus Liberibacter spp., which can be transmitted by grafting and/or the psyllid vectors Diaphorina citri (ACP) and Trioza erytreae (AfCP). Although HLB has not been reported in the Mediterranean Basin to date, both vectors are present, and thus represent a serious threat to the citrus industry in this region. Resistant citrus cultivars or effective therapeutic treatments are not currently available for HLB. Nevertheless, area-wide pest management via coordinated management efforts over large areas has been implemented in Brazil, China and the USA for HLB control. This study proposes an open access flexible methodology to address area-wide management of both HLB vectors in the Mediterranean Basin. Based on a risk-based approach which considers climatic information and other variables that may influence vector introduction and spread, such as conventional, organic, abandoned and residential citrus areas as well as transportation corridors, an area-wide management division in pest management areas (PMAs) is proposed. The size and location of these PMAs were estimated by means of a hierarchical clustering algorithm with spatial constraints whose performance was assessed under different configuration scenarios. This proposal may assist policymakers and the citrus industry of the citrus-growing areas of the Mediterranean Basin in risk management planning in the case of the spread of HLB vectors or a possible introduction of the disease. Additionally, it may be a valuable resource to inform opinion dynamic models, enabling the identification of pivotal factors for the success of control measures., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Galvañ, Bassanezi, Luo, Vanaclocha, Vicent and Lázaro.)

World trade has greatly increased in recent decades, together with a higher risk of introducing non-indigenous pests. Introduction trends show no sign of saturation, and it seems likely that many more species will enter and establish in new territories in the future. A key challenge in analysing pest invasion patterns is the paucity of historical data on pest introductions. A comprehensive dataset of pests' introductions in the EU, including their spatial occurrences, is not currently available and information is scattered across different sources or buried in the scientific literature. Therefore, we collected pests' introduction information (e.g., year, host) from online scientific databases and literature; we then gathered primary spatial data related to the site of first introductions. Finally, we identified the potential pathways of entry for each pest. The dataset contains expert-revised data on 278 pests introduced in the EU between 1999 and 2019, alongside their spatial occurrence and potential pathways of entry, providing a basis to better understand the factors associated with the likelihood of pest introduction. It is important to note that this dataset does not contain the current distribution of the introduced pests, but only records of their first introduction in the EU., (© 2023. Springer Nature Limited.)

In this study, the effect of hydroxypropyl methylcellulose (HPMC) and gum Arabic (GA) edible coatings amended with 0.2% geraniol (GE) were evaluated for the control of brown rot, caused by Monilinia fructicola , on artificially inoculated plums ( Prunus salicina Lindl., cv. Angeleno) stored for 5 weeks at 1 °C. Brown rot is the most important pre- and postharvest fungal disease of stone fruits, causing severe economic losses worldwide. Geraniol is an important constituent of many essential oils that can be obtained as a byproduct from different industrial procedures, such as those of the juice industry. Fruit postharvest quality was also evaluated after 5 and 8 weeks of storage at 1 °C, followed by 3 days at 7 °C plus 5 days at 20 °C, simulating packinghouse, transport, and retail shelf-life conditions, respectively. HPMC coatings containing 0.2% GE reduced the incidence and severity of brown rot by 37.5 and 64.8%, respectively, compared to uncoated fruit after 5 weeks of storage at 1 °C. HPMC-coated plums, with and without GE, showed the highest level of firmness, the lowest change in external peel color parameters (L*, a*, b*, C*, hue), and the lowest flesh bleeding compared to uncoated control and GA-coated samples throughout the entire storage period, which correlated with a higher gas barrier of these coatings without negatively affecting sensory quality. Furthermore, the HPMC-0.2% GE coating provided the highest gloss to coated plums, showing the potential of this coating as a safe and environmentally friendly alternative to conventional fungicides and waxes for brown rot control and quality maintenance of cold-stored plums.

A high intramuscular fat content characterizes Wagyu (WY) cattle breed. Our objective was to compare beef from WY, WY-by-Angus, or Wangus (WN) steers with European, Angus-by-Charolais-Limousine crossbred steers (ACL), considering metabolic biomarkers pre-slaughtering and nutritional characteristics, including health-related indexes of the lipid fraction. The fattening system with olein-rich diets and no exercise restriction included 82 steers, 24 WY, 29 WN, and 29 ACL. The slaughter ages and weights were (median and interquartile range) 38.4 mo.-old (34.9-40.3 mo.) and 840 kg (785-895 kg) for WY; for WN, 30.6 mo. (26.9-36.5 mo.) and 832 kg (802-875 kg), and for ACL steers, 20.3 mo.-old (19.0-22.7 mo.) and 780 kg (715-852 kg). Blood lipid-related metabolites, except for non-esterified fatty acids (NEFA) and low-density level cholesterol (LDL), were higher in WY and WN than in ACL, while glucose was lower in WY and WN. Leptin was higher in WN than in ACL. Pre-slaughtering values of plasma HDL underscored as a possible metabolic biomarker directly related to beef quality. The amino-acid content in beef did not differ among experimental groups, except for more crude protein in ACL. Compared to ACL, WY steers showed higher intramuscular fat in sirloin (51.5 vs. 21.9%) and entrecote (59.6 vs. 27.6%), more unsaturated fatty acids in entrecote (55.8 vs. 53.0%), and more oleic acid in sirloin (46 vs. 41.3%) and entrecote (47.5 vs. 43.3%). Compared to ACL entrecote, WY and WN showed better atherogenic (0.6 and 0.55 vs. 0.69), thrombogenicity (0.82 and 0.92 vs. 1.1), and hypocholesterolemic/hypercholesterolemic index (1.9 and 2.1 vs. 1.7). Therefore, beef's nutritional characteristics depend on breed/crossbred, slaughtering age and cut, with WY and WN entrecote samples showing a healthier lipid fraction.

Citrus black spot (CBS), caused by the fungus Phyllosticta citricarpa, is associated with serious yield and quality losses. The climate suitability of the Mediterranean Basin for CBS development has been long debated. However, CBS has been described in Tunisia. In this study, a generic model was used to simulate potential infections by ascospores and pycnidiospores together with a degree-day model to predict the onset of ascospore release. High-resolution climatic data were retrieved from the ERA5-Land dataset for the citrus-growing regions in the Mediterranean Basin and other locations where CBS is present. In general, the onset of ascospore release was predicted to occur late in spring, but there is no agreement on the adequacy of this empirical model for extrapolation to the Mediterranean Basin. The generic model indicated that infections by ascospores and pycnidiospores would be concentrated mainly in autumn, as well as in spring for pycnidiospores. In contrast to previous studies, the percentage of hours suitable for infection was higher for pycnidiospores than for ascospores. The values obtained with the generic infection model for Tunisia and several CBS-affected locations worldwide were similar to those for other citrus-growing regions in Europe and Northern Africa. These results support previous work indicating that the climate of the Mediterranean Basin is suitable for CBS development., (© 2022. The Author(s).)

Japanese Black (Wagyu) cattle produce high-quality beef. However, whether Wagyu steers can be profitably raised under conditions different than the traditional Japanese ones remains unclear. From 2018 to 2020, we raised 262 Wagyu purebred steers, 103 Wagyu-by-Angus (Wangus) crossbred steers, and 43 Angus-by-European (ACL) crossbred steers on a Spanish farm with high welfare standards and a locally sourced, high-olein diet. Factors and factors' interactions impacting steer growth were analyzed using generalized linear models. ACL steers grew faster than the other two groups, with Wangus showing intermediate fattening and muscle development. Average daily weight gains (kg/day) were 0.916 for Wagyu, 1.046 for Wangus, and 1.293 for ACL during the weaning to growing period, and 0.628 for Wagyu, 0.64 for Wangus, and 0.802 for ACL during the growing to fattening phase. ACL showed the lowest marbling rates. Wagyu and Wangus usually showed higher cholesterol, triglycerides, and high-density lipoprotein than ACL. ACL calves may experience greater stress at weaning, as suggested by higher glucose, lactate, and β-hydroxybutyrate than the other groups. The results suggest that Wagyu and Wangus steers showed adequate growth, health, and metabolic development in this type of production system, with Wagyu purebreds probably being more profitable than Wangus crossbreeds.

Spatial species distribution models often assume isotropy and stationarity, implying that spatial dependence is direction-invariant and uniform throughout the study area. However, these assumptions are violated when dispersal barriers are present. Despite this, the issue of nonstationarity has been little explored in the context of plant health. The objective of this study was to evaluate the influence of barriers in the distribution of Xylella fastidiosa in the demarcated area in Alicante, Spain. Occurrence data from 2018 were analyzed through spatial Bayesian hierarchical models. The stationary model, illustrating a scenario without control interventions or geographical features, was compared with three nonstationary models: a model with mountains as physical barriers, and two models with a continuous and discontinuous perimeter barrier representing hypothetical control interventions. In the stationary model, the posterior mean of the spatial range, as the distance where two observations are uncorrelated, was 4,030 m 95% credible interval (2,907 to 5,564). This distance can be used to define the buffer zone in the demarcated area. The predicted probability of X . fastidiosa presence in the area outside the barrier was 0.46 with the stationary model, whereas it was reduced to 0.29 and 0.36 with the continuous and discontinuous barrier models, respectively. Differences between the discontinuous and continuous barrier models showed that breaks, where no control interventions were implemented, resulted in a higher predicted probability of X . fastidiosa presence in the areas with low sampling intensity. These results may help authorities prioritize the areas for surveillance and disease control.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Pomegranate ( Punica granatum L.) is an emerging crop in Italy and particularly in southern regions, such as Apulia, Basilicata, and Sicily, due to favorable climatic conditions. The crop is affected by several pathogenic fungi, primarily in the field, but also during postharvest phases. The most important postharvest fungal diseases in pomegranate are gray and blue molds, black heart and black spot, anthracnose, dry rot, and various soft rots. The limited number of fungicides allowed for treatment in the field and the lack of postharvest fungicides make it difficult to control latent, quiescent, and incipient fungal infections. Symptomatic pomegranates from southern Italy were sampled and isolated fungi were morphologically and molecularly characterized. The data obtained revealed that various species of Penicillium sensu lato (including Talaromyces genus), Alternaria spp., Coniella granati , and Botrytis cinerea were the principal etiological agents of postharvest pomegranate fruit diseases; other relevant pathogens, although less represented, were ascribable to Aspergillus sect. nigri , Colletotrichum acutatum sensu stricto , and Cytospora punicae . About two thirds of the isolated pathogens were responsible for latent infections. The results obtained may be useful in planning phytosanitary control strategies from the field to storage, so as to reduce yield losses.

Novel pectin-based, antifungal, edible coatings (ECs) were formulated by the addition of natural extracts or essential oils (EOs), and their ability to control green mold (GM), caused by Penicillium digitatum , and preserve postharvest quality of 'Valencia' oranges was evaluated. Satureja montana , Cinnamomum zeylanicum (CN), Commiphora myrrha (MY) EOs, eugenol (EU), geraniol (GE), vanillin, and propolis extract were selected as the most effective antifungal agents against P. digitatum in in vitro assays. Pectin-beeswax edible coatings amended with these antifungals were applied to artificially inoculated oranges to evaluate GM control. ECs containing GE (2 g/kg), EU (4 and 8 g/kg), and MY EO (15 g/kg) reduced disease incidence by up to 58% after 8 days of incubation at 20 °C, while CN (8 g/kg) effectively reduced disease severity. Moreover, ECs formulated with EU (8 g/kg) and GE (2 g/kg) were the most effective on artificially inoculated cold-stored oranges, with GM incidence reductions of 56 and 48% after 4 weeks at 5 °C. Furthermore, ECs containing EU and MY reduced weight loss and maintained sensory and physicochemical quality after 8 weeks at 5 °C followed by 7 days at 20 °C. Overall, ECs with EU were the most promising and could be a good natural, safe, and eco-friendly commercial treatment for preserving orange postharvest quality.

Multiple ovulation and embryo transfer (MOET) systems have been intensively implemented in Japanese Black cattle in Japan and to create Japanese Black herds out of these areas. Environmental conditions influence MOET efficiency. Thus, we describe results of 137 in vivo, non-surgical embryo flushings performed between 2016-2020, in a full-blood Japanese Black herd kept in Spain and the possible effects of heat, year, bull, donor genetic value, and metabolic condition. Additionally, 687 embryo transfers were studied for conception rate (CR) and recipient related factors. A total of 71.3% of viable embryos (724/1015) were obtained (5.3 ± 4.34/flushing). Donor metabolites did not affect embryo production ( p > 0.1), although metabolite differences were observed over the years, and by flushing order, probably related to the donor age. CR was not affected by embryo type (fresh vs. frozen), recipient breed, and whether suckling or not suckling ( p > 0.1). CR decreased significantly with heat (44.3 vs. 49.2%; ( p = 0.042)) and numerically increased with recipient parity and ET-number. Pregnant recipients showed significantly higher levels of cholesterol-related metabolites, glucose, and urea ( p < 0.05). Therefore, adequate MOET efficiency can be achieved under these conditions, and heat stress should be strongly avoided during Japanese Black embryo transfers. Moreover, recipients' metabolites are important to achieve pregnancy, being probably related to better nutrient availability during pregnancy.

Background: Two edible coating (EC) emulsions based on potato starch (F6 and F10) alone or formulated with sodium benzoate (SB, 2% w/w) (F6/SB and F10/SB) were evaluated to maintain postharvest quality of cold-stored 'Fino' lemons and control sour rot on lemons artificially inoculated with Geotrichum citri-aurantii. Previous research showed the potential of these ECs to improve the storability of 'Orri' mandarins and reduce citrus green and blue molds caused by Penicillum digitatum and Penicillium italicum, respectively., Results: The coatings F6/SB and F10/SB significantly reduced sour rot incidence and severity compared to uncoated control samples on lemons incubated at 28 °C for 4 and 7 days. The F6/SB coating reduced weight loss and gas exchange compared to uncoated fruit after 2 and 4 weeks of storage at 12 °C plus a shelf life of 1 week at 20 °C, without adversely affecting the lemon physicochemical quality., Conclusion: Overall, the F6/SB coating formulation, composed of pregelatinized potato starch, glyceryl monostearate, glycerol, emulsifiers and SB, with a total solid content of 5.5%, showed the best results in reducing citrus sour rot and maintaining the postharvest quality of cold-stored 'Fino' lemons. Therefore, it showed potential as a new cost-effective postharvest treatment suitable to be included in integrated disease management programs for citrus international markets with zero tolerance to chemical residues. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Androdioecy (the coexistence of males and hermaphrodites) is a rare mating system for which the evolutionary dynamics are poorly understood. Here, we investigate the cottony cushion scale, Icerya purchasi, one of only three reported cases of androdioecy in insects. In this species, female-like hermaphrodites have been shown to produce sperm and self-fertilize. However, males are ocassionally observed as well. In a large genetic analysis, we show for the first time that, although self-fertilization appears to be the primary mode of reproduction, rare outbreeding events do occur in natural populations, supporting the hypothesis that hermaphrodites mate with males and hence androdioecy is the mating system of I. purchasi. Thus, this globally invasive pest insect appears to enjoy the colonization advantages of a selfing organism while also benefitting from periodic reintroduction of genetic variation through outbreeding with males., (© 2021 The Authors. Evolution published by Wiley Periodicals LLC on behalf of The Society for the Study of Evolution.)

The sulfur-containing salts, classified as food additives, sodium metabisulfite (SMBS), potassium metabisulfite (PMBS), aluminum sulfate (AlS), and aluminum potassium sulfate (AlPS), were evaluated for their activity against Monilinia fructicola , Rhizopus stolonifer , and Geotrichum candidum , the most economically important fungal pathogens causing postharvest disease of stone fruit. In in vitro tests with potato dextrose agar (PDA) Petri dishes amended with different concentrations of the salts (0, 10, 20, 30, 50, and 100 mM), SMBS and PMBS at all concentrations, AlS above 20 mM, and AlPS above 30 mM, completely inhibited the mycelial growth of the three fungi after incubation at 25 °C for up to 10 days. In in vivo primary screenings with artificially inoculated nectarines, aqueous solutions of the four salts reduced the incidence and severity of brown rot (BR) at concentrations of 10 and 50 mM, whereas only AlS and AlPS reduced Rhizopus rot (RR), and none of the salts was effective against sour rot (SR). Solutions at 100 mM were phytotoxic and injured the fruit peel. In small-scale trials, 1 min dip treatments at 20 °C in SMBS or PMBS at 10 mM significantly reduced the incidence and severity of BR after incubation at 20 °C for up to 8 days. Conversely, dips in AlS and AlPS reduced neither BR nor RR. Results highlight the potential of SMBS and PMBS as new nonpolluting tools for the integrated control of BR, but not RR and SR, on stone fruit.

Circular leaf spot (CLS), caused by Plurivorosphaerella nawae , is a serious disease affecting persimmon ( Diospyros kaki ) that is characterized by necrotic lesions on leaves, defoliation, and fruit drop. Under Mediterranean conditions, P. nawae forms pseudothecia in the leaf litter in winter, and ascospores are released in spring, infecting susceptible leaves. Persimmon growers are advised to apply fungicides for CLS control during the period of inoculum availability, which was previously defined based on ascospore counts under the microscope. A model of inoculum availability of P. nawae was developed and evaluated as an alternative to ascospore counts. Leaf litter samples were collected weekly in L'Alcúdia (Spain) from 2010 to 2015. Leaves were soaked and placed in a wind tunnel, and the released ascospores of P. nawae were counted. Hierarchical Bayesian beta regression methods were used to model the dynamics of ascospore production in the leaf litter. The selected model included accumulated degree-days (ADDs) and ADDs taking into account the vapor pressure deficit (ADDvpd) as fixed effects and year as random effect. This model had a mean absolute error of 0.042 and a root mean square error of 0.062. The beta regression model was evaluated in four orchards from 2010 to 2015. Higher accuracy was obtained at the beginning and the end of the ascospore production period, which are the events of interest to schedule fungicide sprays for CLS control in Spain. This same modeling framework can be extended to other fungal plant pathogens whose inoculum dynamics are expressed as proportion data.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Sodium metabisulfite (SMBS), potassium metabisulfite (PMBS), aluminum sulfate (AlS) and aluminum potassium sulfate (AlPS), common sulfur-containing salts used as food additives, were evaluated for their antifungal activity against Penicillium digitatum, Penicillium italicum and Geotrichum citri-aurantii, the most economically important pathogens causing postharvest diseases of citrus fruits. In vitro radial mycelial growth was measured on potato dextrose agar (PDA) Petri dishes amended with five different concentrations of the salts (10, 20, 30, 50, 100 mM) after 7 d of incubation at 25 °C. SMBS and PMBS at all concentrations, and AIS and AIPS above 20 mM, completely inhibited the growth of these fungi. The curative antifungal activity of the four salts to control citrus green (GM) and blue (BM) molds and sour rot (SR) was evaluated on 'Valencia' oranges artificially inoculated in rind wounds with P. digitatum, P. italicum and G. citri-aurantii, respectively. In vivo primary screenings showed no significant antifungal activity of AlS and AlPS to control the three diseases at any dose tested, but SMBS and PMBS reduced the incidence and severity of GM, BM and SR at various concentrations. Effective salts and concentrations were selected for in vivo dip treatments in small-scale trials. Dips at room temperature (20 °C) in SMBS and PMBS at 20 and 50 mM for 60 or 120 s significantly reduced the incidence and severity of GM and BM, with PMBS at 50 mM for 120 s the most effective treatment. Conversely, dips in SMBS and PMBS at 50 mM for 60 or 120 s did not reduce SR incidence and severity. SMBS and PMBS treatments are potentially new tools to be included in reduced-risk non-polluting strategies to control Penicillium diseases, but not SR, on citrus fruits., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The incidence of peach powdery mildew (PPM) on fruit was monitored in commercial peach orchards to i) describe the disease progress in relation to several environmental parameters and ii) establish an operating threshold to initiate a fungicide spray program based on accumulated degree-day (ADD) data. A beta-regression model for disease incidence showed a substantial contribution of the random effects orchard and year, whereas relevant fixed effects corresponded to ADD, wetness duration, and ADD considering vapor pressure deficit and rain. When beta-regression models were fitted for each orchard and year considering only ADD, disease onset was observed at 242 ± 13 ADD and symptoms did not develop further after 484 ± 42 ADD. An operating threshold to initiate fungicide applications was established at 220 ADD, coinciding with a PPM incidence in fruit around 0.05. A validation was further conducted by comparing PPM incidence in i) a standard, calendar-based program, ii) a program with applications initiated at 220 ADD, and iii) a nontreated control. A statistically relevant reduction in disease incidence in fruit was obtained with both fungicide programs, from 0.244 recorded in the control to 0.073 with the 220-ADD alert program, and 0.049 with the standard program. The 220-ADD alert program resulted in 33% reduction in fungicide applications.

The plant-pathogenic bacterium Xylella fastidiosa was first reported in Europe in 2013, in the province of Lecce, Italy, where extensive areas were affected by the olive quick decline syndrome, caused by the subsp. pauca . In Alicante, Spain, almond leaf scorch, caused by X. fastidiosa subsp. multiplex , was detected in 2017. The effects of climatic and spatial factors on the geographic distribution of X. fastidiosa in these two infested regions in Europe were studied. The presence/absence data of X. fastidiosa in the official surveys were analyzed using Bayesian hierarchical models through the integrated nested Laplace approximation (INLA) methodology. Climatic covariates were obtained from the WorldClim v.2 database. A categorical variable was also included according to Purcell's minimum winter temperature thresholds for the risk of occurrence of Pierce's disease of grapevine, caused by X. fastidiosa subsp. fastidiosa . In Alicante, data were presented aggregated on a 1 km grid (lattice data), where the spatial effect was included in the model through a conditional autoregressive structure. In Lecce, data were observed at continuous locations occurring within a defined spatial domain (geostatistical data). Therefore, the spatial effect was included via the stochastic partial differential equation approach. In Alicante, the pathogen was detected in all four of Purcell's categories, illustrating the environmental plasticity of the subsp. multiplex . Here, none of the climatic covariates were retained in the selected model. Only two of Purcell's categories were represented in Lecce. The mean diurnal range ( bio2 ) and the mean temperature of the wettest quarter ( bio8 ) were retained in the selected model, with a negative relationship with the presence of the pathogen. However, this may be due to the heterogeneous sampling distribution having a confounding effect with the climatic covariates. In both regions, the spatial structure had a strong influence on the models, but not the climatic covariates. Therefore, pathogen distribution was largely defined by the spatial relationship between geographic locations. This substantial contribution of the spatial effect in the models might indicate that the current extent of X. fastidiosa in the study regions had arisen from a single focus or from several foci, which have been coalesced., (Copyright © 2020 Cendoya, Martínez-Minaya, Dalmau, Ferrer, Saponari, Conesa, López-Quílez and Vicent.)

Xylella fastidiosa is the causal agent of plant diseases that cause massive economic damage. In 2013, a strain of the bacterium was, for the first time, detected in the European territory (Italy), causing the Olive Quick Decline Syndrome. We simulate future spread of the disease based on climatic-suitability modeling and radial expansion of the invaded territory. An economic model is developed to compute impact based on discounted foregone profits and losses in investment. The model projects impact for Italy, Greece, and Spain, as these countries account for around 95% of the European olive oil production. Climatic suitability modeling indicates that, depending on the suitability threshold, 95.5 to 98.9%, 99.2 to 99.8%, and 84.6 to 99.1% of the national areas of production fall into suitable territory in Italy, Greece, and Spain, respectively. For Italy, across the considered rates of radial range expansion the potential economic impact over 50 y ranges from 1.9 billion to 5.2 billion Euros for the economic worst-case scenario, in which production ceases after orchards die off. If replanting with resistant varieties is feasible, the impact ranges from 0.6 billion to 1.6 billion Euros. Depending on whether replanting is feasible, between 0.5 billion and 1.3 billion Euros can be saved over the course of 50 y if disease spread is reduced from 5.18 to 1.1 km per year. The analysis stresses the necessity to strengthen the ongoing research on cultivar resistance traits and application of phytosanitary measures, including vector control and inoculum suppression, by removing host plants., Competing Interests: The authors declare no competing interest., (Copyright © 2020 the Author(s). Published by PNAS.)

Background: Small RNAs regulate a wide variety of processes in plants, from organ development to both biotic and abiotic stress response. Being master regulators in genetic networks, their biogenesis and action is a fundamental aspect to characterize in order to understand plant growth and development. Three main gene families are critical components of RNA silencing: DICER-LIKE (DCL), ARGONAUTE (AGO) and RNA-DEPENDENT RNA POLYMERASE (RDR). Even though they have been characterized in other plant species, there is no information about these gene families in Citrus sinensis, one of the most important fruit species from both economical and nutritional reasons. While small RNAs have been implicated in the regulation of multiple aspects of plant growth and development, their role in the abscission process has not been characterized yet., Results: Using genome-wide analysis and a phylogenetic approach, we identified a total of 13 AGO, 5 DCL and 7 RDR genes. We characterized their expression patterns in root, leaf, flesh, peel and embryo samples using RNA-seq data. Moreover, we studied their role in fruit abscission through gene expression analysis in fruit rind compared to abscission zone from samples obtained by laser capture microdissection. Interestingly, we determined that the expression of several RNA silencing factors are down-regulated in fruit abscission zone, being particularly represented gene components of the RNA-dependent DNA Methylation pathway, indicating that repression of this process is necessary for fruit abscission to take place in Citrus sinensis., Conclusions: The members of these 3 families present characteristic conserved domains and distinct expression patterns. We provide a detailed analysis of the members of these families and improved the annotation of some of these genes based on RNA-seq data. Our data suggests that the RNA-dependent DNA Methylation pathway is involved in the important fruit abscission process in C. sinensis.

A large amount of GRAS (generally recognized as safe) salts and concentrations were evaluated in in vitro tests (inhibition of mycelial growth on PDA dishes) against Lasiodiplodia theobromae, the causal agent of citrus Diplodia stem-end rot. Ammonium carbonate (AC, 0.2%), potassium sorbate (PS, 2.0%), potassium carbonate (PC, 0.2%), sodium methylparaben (SMP, 0.1%), sodium ethylparaben (SEP, 0.1%), sodium benzoate (SB, 2.0%), and potassium silicate (PSi, 2.0%) were selected as the most effective. Disease control ability of edible composite coatings formulated with hydroxypropyl methylcellulose (HPMC), beeswax (BW), and these selected antifungal GRAS salts was assessed in in vivo experiments with 'Ortanique' mandarins and 'Barnfield' oranges artificially inoculated with L. theobromae. Coatings containing 2% PS, 0.1% SEP, or 2% SB were the most effective reducing disease severity (up to 50% reduction) and were also applied to non-inoculated and cold-stored 'Barnfield' oranges to determine their effect on postharvest fruit quality. After periods of 21 and 42 d at 5 °C followed by 7 d of shelf life at 20 °C, coatings containing SEP and SB significantly reduced weight loss and did not adversely affect the physicochemical quality attributes (firmness, soluble solid content, titratable acidity, and ethanol and acetaldehyde content) and sensory flavor with respect to uncoated control fruit. Although the internal gas concentration (CO 2 level) of coated fruit increased, the coatings did not induce off-flavors., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The red palm weevil (RPW) Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) is threatening the palm family worldwide, causing important economic losses. Current tactics to manage the weevil are largely based on chemical control, although the use of pesticides is hampered by several environmental constraints. Since the first introduction of RPW in Spain in 1996 and during its progressive spread around the Mediterranean basin, the number of reports of natural infection of RPW populations by entomopathogenic fungi (EPF) has been rising for 15 years, and this rise could support a pest-mediated EPF spread. To challenge this hypothesis, we assessed the usefulness of the region of elongation factor 1-alpha (EF1-α), Bloc nuclear intergenic region (Bloc) and inter simple sequence repeat (ISSR) markers, alone or in combination, to infer the relationships among Mediterranean Beauveria and Metarhizium strains isolated from the RPW. Second, the effect of abiotic factors, such as temperature, humidity and UV-B radiation, on the germination and growth of these EPFs strains as a function of their genealogy and geographic origin were determined. Finally, the pathogenicity of strains from different genetic clades was evaluated against larvae and adults of R. ferrugineus. The phylogenetic analysis based on the EF-1α gene identified eight different sequences among 24 fungal isolates of four fungal species. Similar clades were clustered when Bloc and ISSR analyses were performed. The results showed that strains of different origins were clustered in the same clade, and this outcome could be explained by an RPW-mediated EPF spread that was also influenced by time, geographical and other RPW related factors. Neither the response to abiotic factors nor virulence to RPW larvae and adults were related to the sequence type, with all B. bassiana strains well adapted to Mediterraneam climatic conditions. Taken together, these findings may help to select the best strain for RPW management., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: A new Beauveria bassiana-based attract and infect device (AID) to control Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) was developed. The virulence and persistence of the fungal formulation used in the AID were evaluated in the laboratory. Semi-field and field trials were carried out to validate the results and establish the potential of this device as a control tool., Results: In laboratory conditions, a 50% lethal time (LT 50 ) of 4.33 days was obtained when adults (7-10 days old) were exposed to the inoculation tunnel (IT) containing 1 × 10 10 conidia g -1 in an oil-based fungal formulation. This formulation maintained conidium viability at 50% for up to 2 months. Moreover, when adults were exposed to 2.5-month field-aged ITs, mortality still reached 50% 40 days after exposure. In addition, no differences were observed between ITs aged in early spring and those aged in summer, suggesting that the fungal formulation is not strongly affected by environmental factors in Mediterranean basin conditions. Semi-field assays showed that the device allowed an easy transit of weevils through the IT, which were effectively attracted and infected. Using the AIDs in 4-ha plot field trials, a reduction of >50% in the percentage of infested sentinel palms was obtained., Conclusion: Based on the results obtained in terms of the efficacy and persistence of this new AID in the field and its potential in reducing R. ferrugineus populations and palm infestation, this device could become a key tool for the management of R. ferrugineus. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)