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Silent effect of the fungicide pyraclostrobin on the larval exposure of the non-target organism Africanized Apis mellifera and its interaction with the pathogen Nosema ceranae in adulthood Texto completo
2020
Tadei, Rafaela | Menezes-Oliveira, Vanessa B. | Silva-Zacarin, Elaine C.M.
The frequent exposure of bees to a wide variety of fungicides, on crops where they forage, can be considered a stressor factor for these pollinators. The organisms are exposed both to the fungicide active ingredients and to the adjuvants of commercial formulations. All these ingredients are brought to the hive by bee foragers through contaminated pollen and nectar, thus exposing also immature individuals during larval phase. This work aimed to compare the effects of larval exposure to the fungicide pyraclostrobin (active ingredient and commercial formulation) and its influence on the cytotoxicity to midguts in adults, which were inoculated with the Nosema ceranae spores in the post-emergence stage. Under laboratory conditions, Apis mellifera larvae received an artificial diet containing fungicide solution from the third to the sixth day of the feeding phase. One-day-old adult workers ingested 100,000 infectious N. ceranae spores mixed in sucrose solution. Effects on midgut were evaluated through cellular biomarkers of stress and cell death. The exposure to the fungicide (active ingredient and commercial formulation) did not affect the larval post-embryonic development and survival of adult bees. However, this exposure induced cytotoxicity in the cells of the midgut, showed by the increase in DNA fragmentation and alteration in the HSP70 immunolabeling pattern. Without the pathogen, the midgut cytotoxic effects and HSP70 immunolabeling of the organisms exposed to the commercial formulation were lower when compared to the exposure to its active ingredient. However, in the presence of the pathogen, the cytotoxic effects of the commercial formulation to the adult bees’ midgut were potentialized. The pathogen N. ceranae increased the damage to the intestinal epithelium of adult bees. Thus, realistic doses of pyraclostrobin present in beebread consumed by larvae can affect the health and induce physiological implications to the midgut functions of the adult bees.
Mostrar más [+] Menos [-]Liver-derived exosome-laden lncRNA MT1DP aggravates cadmium-induced nephrotoxicity Texto completo
2020
Gao, Ming | Dong, Zheng | Sun, Jinfang | Liu, Wei | Xu, Ming | Li, Changying | Zhu, Pan | Yang, Xingfeng | Shang, Xiaohong | Wu, Yongning | Liu, Sijin
Cadmium (Cd) is a well-characterized toxic heavy metal which could cause severe kidney injury. However, currently the knowledge of Cd toxicity towards kidney is still insufficient. Our previous data has identified that MT1DP (metallothionein 1D pseudogene) could promote Cd-induced detrimental effects on hepatocytes. Herein, we further found that MT1DP was also an important intermediate to aggravate Cd-induced nephrotoxicity. Through analyzing the data of 100 residents from Cd-contaminated area in Southern China, we found that the blood MT1DP levels correlated to the urine Cd content and the extent of nephrotoxicity. Although MT1DP was predominantly induced by hepatocytes in the liver, liver-secreted MT1DP was found to be packaged into extracellular cargoes: exosomes, by which MT1DP was delivered into circulation and thereafter targeted kidney cells. Furthermore, exosome-laden MT1DP worsened Cd-induced nephrotoxicity, as evidenced in both Cd-poisoned individuals and in vitro cells. Moreover, MT1DP was found to reinforce Cd-induced toxicity in kidney cells by indirectly breaking the equilibrium between the pro-apoptotic and anti-apoptotic effects conducted by BAX and Bcl-xL, respectively. Collectively, our data unveiled that hepatocyte-derived MT1DP depends on the delivery of exosomes to wreak considerable havoc in Cd nephrotoxicity. This study offers new insights into the molecular mechanisms of Cd-induced kidney injury.
Mostrar más [+] Menos [-]Effective treatment of levofloxacin wastewater by an electro-Fenton process with hydrothermal-activated graphite felt as cathode Texto completo
2020
Liu, Jia-Ming | Ji, Zhi-Yong | Shi, Ya-Bin | Yuan, Peng | Guo, Xiao-Fu | Zhao, Li-Ming | Li, Shuming | Li, Hong | Yuan, Jun-Sheng
The performance of the cathode significantly affects the ability of the electro-Fenton (EF) process to degrade chemicals. In this study, a simple method to modify the graphite felt (GF) cathode was proposed, i.e. oxidizing GF by hydrothermal treatment in nitric acid. The surface physical and electrochemical properties of modified graphite felt were characterized by several techniques: scanning electron microscope (SEM), water contact angle, X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and linear scanning voltammetry (LSV). Compared with an unmodified GF (GF-0), the oxygen reduction reaction (ORR) activity of a modified GF was significantly improved due to the introduction of more oxygen-containing functional groups (OGs). Furthermore, the results showed that GF was optimally modified after 9 h (GF-9) of treatment. As an example, the H₂O₂ generation by GF-9 was 2.26 times higher than that of GF-0. After optimizing the process parameters, which include the initial Fe²⁺ concentration and current density, the apparent degradation rate constant of levofloxacin (LEV) could reach as high as 0.40 min⁻¹. Moreover, the total organic carbon (TOC) removal rate and mineralization current efficiency (MCE) of the modified cathode were much higher than that of the GF-0. Conclusively, GF-9 is a promising cathode for the future development in organic pollutant removal via EF.
Mostrar más [+] Menos [-]Incorporation of solar-thermal energy into a gasification process to co-produce bio-fertilizer and power Texto completo
2020
Ansari, Shahid H. | Ahmed, Ashfaq | Razzaq, Abdul | Hildebrandt, Diane | Liu, Xinying | Park, Young-Kwon
Incorporation of solar-thermal energy into a gasification process to co-produce bio-fertilizer and power Texto completo
2020
Ansari, Shahid H. | Ahmed, Ashfaq | Razzaq, Abdul | Hildebrandt, Diane | Liu, Xinying | Park, Young-Kwon
Biomass integrated gasification combined cycle (IGCC) is attracting increased interest because it can achieve high system energy efficiency (>50%), which is predicted to increase with the increase in the solar share in biomass IGCC. This study evaluated the potential of crop residues numerically for the co-production of power and bio-fertilizer using ASPEN Plus® simulation software. The results showed that the gas yield increases with increasing temperature and decreasing pressure while the yield of bio-fertilizer is dependent on the biomass composition. The biomass with a low ash content produces high bio-fertilizer at the designated gasification temperature. The IGCC configuration conserves more energy than a directly-fired biomass power plant. In addition, the solar-assisted IGCC attains a higher net electricity output per unit of crop residue feed and achieves net thermal efficiencies of around 53%. The use of such hybrid systems offer the potential to produce 0.55 MW of electricity per unit of solar-thermal energy at a relatively low cost. The ASPEN Plus model predicted that the solar biomass-based IGCC set up is more efficient in increasing the power generation capacity than any other conversion system. The results showed that a solar to electricity efficiency of approximately 55% is achievable with potential improvements. This work will contribute for the sustainable bioenergy production as the relationship between energy production and biomass supplies very important to ensure the food security and environmental sustainability.
Mostrar más [+] Menos [-]Incorporation of solar-thermal energy into a gasification process to co-produce bio-fertilizer and power Texto completo
2020
Ansari, Shahid H | Ahmed, Ashfaq | Razzaq, Abdul | Hildebrandt, Diane | Liu, Xinying | Park, Young-Kwon
Gene expression response of the alga Fucus virsoides (Fucales, Ochrophyta) to glyphosate solution exposure Texto completo
2020
Gerdol, Marco | Visintin, Andrea | Kaleb, Sara | Spazzali, Francesca | Pallavicini, Alberto | Falace, Annalisa
Fucus virsoides is an ecologically important canopy-forming brown algae endemic to the Adriatic Sea. Once widespread in marine coastal areas, this species underwent a rapid population decline and is now confined to small residual areas. Although the reasons behind this progressive disappearance are still a matter of debate, F. virsoides may suffer, like other macroalgae, from the potential toxic effects of glyphosate-based herbicides.Here, through a transcriptomic approach, we investigate the molecular basis of the high susceptibility of this species to glyphosate solution, previously observed at the morphological and eco-physiological levels. By simulating runoff event in a factorial experiment, we exposed F. virsoides to glyphosate (Roundup® 2.0), either alone or in association with nutrient enrichment, highlighting significant alterations of gene expression profiles that were already visible after three days of exposure. In particular, glyphosate exposure determined the near-complete expression shutdown of several genes involved in photosynthesis, protein synthesis and stress response molecular pathways. Curiously, these detrimental effects were partially mitigated by nutrient supplementation, which may explain the survival of relict population in confined areas with high nutrient inputs.
Mostrar más [+] Menos [-]High temporal resolution measurements of ammonia emissions following different nitrogen application rates from a rice field in the Taihu Lake Region of China Texto completo
2020
Yang, Wenliang | Que, Huali | Wang, Shuwei | Zhu, Anning | Zhang, Yujun | He, Ying | Xin, Xiuli | Zhang, Xianfeng | Ding, Shijie
Ammonia emission is one of the dominant pathways of nitrogen fertilizer loss from rice fields in China. It is difficult to measure ammonia emissions by high-frequency sampling with the chamber methods widely used in China, which is of great significance for investigating the environmental effects on the ammonia emissions. The chamber methods also can not accurately determine the ammonia emissions. In this study, the backward Lagrangian stochastic dispersion model, with ammonia concentrations continuously measured by the open-path tunable diode laser absorption spectroscopy technique, was used to determine ammonia emissions from a rice field after fertilizer application at excessive (270 kg N ha⁻¹) and appropriate (210 kg N ha⁻¹) rates in the Taihu Lake Region of China. High temporal resolution measurements of ammonia emissions revealed that high intraday fluctuations of ammonia emissions were significantly affected by the meteorological conditions. Multiple regression analysis showed a dominant solar radiation dependence of intraday ammonia emission cycles, especially during the rice panicle formation stage. The NH₄⁺-N concentrations of the surface water of the rice field were found to be the decisive factor that influenced interday dynamics of ammonia emissions. Accurate quantifications of ammonia emissions indicated that the total ammonia losses under appropriate nitrogen application rate were 27.4 kg N ha⁻¹ during the rice tillering stage and 11.2 kg N ha⁻¹ during the panicle formation stage, which were 29.4% and 17.0% less than those under traditional excessive nitrogen application rate used by the local farmers, respectively. The ammonia loss proportions during the rice panicle formation stage were significantly lower than those of the tillering stage, which might be due to different nitrogen application rates and environmental effects during the two stages. This study indicated that the open-path tunable diode laser absorption spectroscopy technique could facilitate the investigation of high temporal resolution dynamic of ammonia emissions from farmland and the environmental influence on the ammonia emissions.
Mostrar más [+] Menos [-]The interference of nonylphenol with bacterial cell-to-cell communication Texto completo
2020
Jayaprada, Thilini | Hu, Jingming | Zhang, Yunyun | Feng, Huajun | Shen, Dongsheng | Geekiyanage, Sudarshanee | Yao, Yanlai | Wang, Meizhen
The interference of nonylphenol (NP) with humans and animals, especially in hormone systems, has been well-studied. There is rarely any record of its effect on bacteria, which dominate in various environments. In our study, we employed Pseudomonas aeruginosa PAO1 as a model microorganism and took its common lifestyle biofilm, mainly regulated by quorum sensing (QS), as a cut-in point to investigate the effect of NP (1, 5, 10 mg L⁻¹) on bacteria. The results showed that more than 5 mg L⁻¹ of NP did interfere with biofilm formation and affected bacterial QS. In detail, the LasI/R circuit, but not the RhlI/R circuit, was considerably obstructed. The decrease in lasI and lasR expression resulted in a significant reduction in N-3-oxo-dodecanoyl homoserine lactone (3OC₁₂-HSL) signals and the downstream production of elastases. Docking results indicated the binding of NP with LasR protein, simulating the binding of 3OC₁₂-HSL with LasR protein, which explained the obstruction of the LasIR circuit. We concluded that NP competed with 3OC₁₂-HSL and blocked 3OC₁₂-HSL binding with the LasR protein, resulting in a direct interference in bacterial biofilm formation. This is the first report of NP interference with bacterial signaling, which is not only helpful to understand the effect of NP on various ecosystems, but is also beneficial to enrich our knowledge of inter-kingdom communication.
Mostrar más [+] Menos [-]Heavy metal accumulation and genotoxic effects in levant vole (Microtus guentheri) collected from contaminated areas due to mining activities Texto completo
2020
Turna Demir, Fatma | Yavuz, Mustafa
Heavy metal contamination is a serious environmental problem commonly monitored in various organisms. Small wild rodents are ideal biological monitors to show the extent of environmental pollution. The aim of this study was to evaluate the adverse effects of marble and stone quarries on the Levant vole, Microtus guentheri, inhabiting some polluted sites. In this context, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was used to analyze distribution of thirteen heavy metals (Fe, Al, Zn, Cu, Cr, Mn, Ni, B, Pb, As, Co, Cd, and Hg) in the organs (skins, bones, muscles, livers and kidneys) of the biological specimens, and the comet assay revealed DNA damage in blood lymphocytes for the first time. This study was conducted at close to the marble and stone quarries at Korkuteli, Antalya-Turkey during spring, summer, autumn (2017) and winter (2018) seasons. In spring and summer, genetic damage in blood lymphocytes from all polluted sites (sites 1–5) was significantly higher than that of controls, while in autumn it was higher in samples from three sites (sites 3–5). In terms of heavy metal distribution in organs, we found depositions of Fe, Al, Zn, Ni, Mn, Cr, Co, As and Pb primarily in the skin with its derivatives, Cu and Cd deposits in the kidney, Cu, Cd and B deposits in the liver, and As and Pb depositions in the bones. The study shows that certain organs (especially skin with its derivatives) and blood lymphocytes of Levant vole can be used as ideal indicators of heavy metal pollution. Our results suggest that the Korkuteli area could already be under the threat of heavy metal pollution.
Mostrar más [+] Menos [-]Health risk assessments based on polycyclic aromatic hydrocarbons in freshwater fish cultured using food waste-based diets Texto completo
2020
Man, Yu Bon | Mo, Wing Yin | Zhang, Feng | Wong, Ming Hung
Two farmed freshwater fish species Nile tilapia (Oreochromis niloticus) and jade perch (Scortum barcoo) were cultured with food waste-based diets and compared with commercial formulated control diet for a period of six months. Sixteen priority polycyclic aromatic hydrocarbons (PAHs) in the diets and cultured fish meat were tested by gas chromatography–mass spectrometry. No significant differences of ∑PAHs were observed between Nile tilapia and jade perch fed with food waste-based diets and control diet (p > 0.05). However, there were significantly higher concentration of ∑PAHs in market fish compared with the same species of fish fed by food waste-based diets (p < 0.05). Thus, the food waste-based diets have a potential to lower the PAH concentrations in farmed fish when compared with market fish. Based on the PAH concentrations, a human health risk assessment was made. The results indicated there were no non-cancer and very low cancer risks of consuming fish cultured with food waste-based diets at the 95th centile (Nile tilapia: hazard index (HI adult) = 0.343 × 10−3, HI children = 0.614 × 10−3 and cancer risk value = 0.943 × 10−6; jade perch: HI adult = 0.456 × 10−3, HI children = 0.814 × 10−3 and cancer risk value = 0.291 × 10−6). In general, the fish fed with food waste-based diets were unlikely to cause adverse health effects, based on the concentrations of PAHs. There is great potential for using food waste-based diets as an alternative to commercial feeds for cultivating freshwater fish.
Mostrar más [+] Menos [-]IPM-recommended insecticides harm beneficial insects through contaminated honeydew Texto completo
2020
Calvo-Agudo, Miguel | González-Cabrera, Joel | Sadutto, Daniele | Picó, Yolanda | Urbaneja, Alberto | Dicke, Marcel | Tena, Alejandro
IPM-recommended insecticides harm beneficial insects through contaminated honeydew Texto completo
2020
Calvo-Agudo, Miguel | González-Cabrera, Joel | Sadutto, Daniele | Picó, Yolanda | Urbaneja, Alberto | Dicke, Marcel | Tena, Alejandro
The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insecticides pymetrozine and flonicamid, which are commonly used in Integrated Pest Management programs, were present in honeydew excreted by the mealybug Planococcus citri. However, only pymetrozine was detected in honeydew excreted by the whitefly Aleurothixus floccosus. Toxicological studies demonstrated that honeydew excreted by mealybugs feeding on trees treated either with flonicamid or pymetrozine increased the mortality of the hoverfly Sphaerophoria rueppellii, but did not affect the parasitic wasp Anagyrusvladimiri. Honeydew contaminated with flonicamid was more toxic for the hoverfly than that contaminated with pymetrozine. Collectively, our data demonstrate that systemic insecticides commonly used in IPM programs can contaminate honeydew and kill beneficial insects that feed on it, with their toxicity being dependent on the active ingredient and hemipteran species that excretes the honeydew.
Mostrar más [+] Menos [-]IPM-recommended insecticides harm beneficial insects through contaminated honeydew
2020
Calvo-Agudo, Miguel | González-Cabrera, Joel | Sadutto, Daniele | Picó, Yolanda | Urbaneja, Alberto | Dicke, Marcel | Tena, Alejandro
The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insecticides pymetrozine and flonicamid, which are commonly used in Integrated Pest Management programs, were present in honeydew excreted by the mealybug Planococcus citri. However, only pymetrozine was detected in honeydew excreted by the whitefly Aleurothixus floccosus. Toxicological studies demonstrated that honeydew excreted by mealybugs feeding on trees treated either with flonicamid or pymetrozine increased the mortality of the hoverfly Sphaerophoria rueppellii, but did not affect the parasitic wasp Anagyrus vladimiri. Honeydew contaminated with flonicamid was more toxic for the hoverfly than that contaminated with pymetrozine. Collectively, our data demonstrate that systemic insecticides commonly used in IPM programs can contaminate honeydew and kill beneficial insects that feed on it, with their toxicity being dependent on the active ingredient and hemipteran species that excretes the honeydew. Insecticides recommended in Integrated Pest Management programs reach honeydew and kill beneficial insects that feed on it.
Mostrar más [+] Menos [-]IPM-recommended insecticides harm beneficial insects through contaminated honeydew Texto completo
2020
Calvo-Agudo, Miguel | González-Cabrera, Joel | Sadutto, Daniele | Picó, Yolanda | Urbaneja, Alberto | Dicke, Marcel | Tena, Alejandro
The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insecticides pymetrozine and flonicamid, which are commonly used in Integrated Pest Management programs, were present in honeydew excreted by the mealybug Planococcus citri. However, only pymetrozine was detected in honeydew excreted by the whitefly Aleurothixus floccosus. Toxicological studies demonstrated that honeydew excreted by mealybugs feeding on trees treated either with flonicamid or pymetrozine increased the mortality of the hoverfly Sphaerophoria rueppellii, but did not affect the parasitic wasp Anagyrus vladimiri. Honeydew contaminated with flonicamid was more toxic for the hoverfly than that contaminated with pymetrozine. Collectively, our data demonstrate that systemic insecticides commonly used in IPM programs can contaminate honeydew and kill beneficial insects that feed on it, with their toxicity being dependent on the active ingredient and hemipteran species that excretes the honeydew.
Mostrar más [+] Menos [-]IPM-recommended insecticides harm beneficial insects through contaminated honeydew Texto completo
2020
Calvo-Agudo, Miguel | González-Cabrera, Joel | Sadutto, Daniele | Picó, Yolanda | Urbaneja, Alberto | Dicke, Marcel | Tena, Alejandro
The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insecticides pymetrozine and flonicamid, which are commonly used in Integrated Pest Management programs, were present in honeydew excreted by the mealybug Planococcus citri. However, only pymetrozine was detected in honeydew excreted by the whitefly Aleurothixus floccosus. Toxicological studies demonstrated that honeydew excreted by mealybugs feeding on trees treated either with flonicamid or pymetrozine increased the mortality of the hoverfly Sphaerophoria rueppellii, but did not affect the parasitic wasp Anagyrus vladimiri. Honeydew contaminated with flonicamid was more toxic for the hoverfly than that contaminated with pymetrozine. Collectively, our data demonstrate that systemic insecticides commonly used in IPM programs can contaminate honeydew and kill beneficial insects that feed on it, with their toxicity being dependent on the active ingredient and hemipteran species that excretes the honeydew. Insecticides recommended in Integrated Pest Management programs reach honeydew and kill beneficial insects that feed on it.
Mostrar más [+] Menos [-]IPM-recommended insecticides harm beneficial insects through contaminated honeydew Texto completo
2020
Calvo-Agudo, Miguel | González-Cabrera, Joel | Sadutto, D. | Picó, Yolanda | Urbaneja, Alberto | Dicke, Marcel | Tena, Alejandro | CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) | Generalitat Valenciana | Ministerio de Economía y Competitividad (España)
The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insecticides pymetrozine and flonicamid, which are commonly used in Integrated Pest Management programs, were present in honeydew excreted by the mealybug Planococcus citri. However, only pymetrozine was detected in honeydew excreted by the whitefly Aleurothixus floccosus. Toxicological studies demonstrated that honeydew excreted by mealybugs feeding on trees treated either with flonicamid or pymetrozine increased the mortality of the hoverfly Sphaerophoria rueppellii, but did not affect the parasitic wasp Anagyrus vladimiri. Honeydew contaminated with flonicamid was more toxic for the hoverfly than that contaminated with pymetrozine. Collectively, our data demonstrate that systemic insecticides commonly used in IPM programs can contaminate honeydew and kill beneficial insects that feed on it, with their toxicity being dependent on the active ingredient and hemipteran species that excretes the honeydew. | This research was partially funded by an Instituto Nacional de Investigaciones Agrarias (INIA) (Project RTA2017-00095) and the Conselleria d’Agricultura, Pesca i Alimentació de la Generalitat Valenciana. J.G.C was supported by the Spanish Ministry of Economy and Competitiveness, Ramón y Cajal Program (RYC-2013-13834) and M.C.A was recipient of grant from INIA (CPD2016-0085).
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