Estudio de los factores del estilo de vida, genéticos y epigenéticos, que influyen en la obesidad y enfermedades relacionadas en población mediterránea

La obesidad ha sido catalogada como la epidemia del siglo XXI, cuya prevalencia a nivel mundial ha ido aumentado en las últimas décadas. La obesidad se define como una acumulación anormal o excesiva de grasa que puede ser perjudicial para la salud. Además de ser considerada una enfermedad, es un factor de riesgo para otras patologías. El origen de la obesidad es multifactorial, y se produce como resultado de la combinación de los efectos genéticos, ambientales y sus interacciones. Los factores ambientales son factores modificables que pueden considerarse de estilo de vida, y los factores genéticos son considerados no modificables teniendo un papel importante, entre el 40-80% en la susceptibilidad del desarrollo de obesidad. Por este motivo el objetivo de esta tesis doctoral fue estudiar la influencia de los factores ambientales y genéticos en la obesidad y enfermedades relacionadas en una población de casos de obesidad y controles del estudio OBENUTIC (Obesidad, Nutrición y Tecnologías de la Información y Comunicación) y en población de riesgo cardiovascular, estudio PREDIMED (PREvención Dieta MEDiterránea). Así como analizar el nivel de expresión de genes relacionados con la obesidad y el sueño tras la temporalidad en pacientes de población general. Los resultados más relevantes mostraron fuertes asociaciones entre el estilo de vida y la obesidad, siendo la realización de actividad física, el patrón de sueño y la calidad de la dieta los factores más importantes en la obesidad y parámetros asociados. Respecto a los factores genéticos, se observó que polimorfismos en genes receptores del sabor se relacionaron con la percepción de determinados sabores y con el consumo de alimentos característicos. A su vez, estos polimorfismos genéticos, como el rs713598 del gen TAS2R38 y el rs35874116 del gen TAS1R2 mostraron asociación con parámetros de obesidad. Por otra parte, genes asociados con el ciclo circadiano (rs4580704 del gen CLOCK y rs1982350 del gen BMAL1) mostraron asociación con el IMC y el nivel de glucosa plasmático. Variantes genéticas de genes implicados en la ingesta dietética, metabolismo lipídico y glucídico como el rs9939606 del gen FTO, rs3802177 del gen SLC3048 o rs708272 del gen CETP, se asociaron con fenotipos de obesidad. Además, en pacientes del estudio PREDIMED, polimorfismos en microRNAs (rs895819 en el pre-miR-27a, rs11614913 en miRNA196a2 y rs2292832 en miRNA-149) han mostrado una asociación con el consumo de alimentos. Algunos de los polimorfismos analizados, presentaron modulaciones ambientales en relación con la obesidad o parámetros asociados. Por otra parte, se observaron cambios en el nivel de expresión tras la temporalidad y se observó mayores niveles de expresión en los genes FTO, CLOCK, PER1, CD36 y VRK2 en los individuos que habían mostrado una pérdida de peso significativa. Estos resultados, ponen de manifiesto la importancia del estudio de los factores del estilo de vida, factores genéticos y de expresión génica, así como su interacción, cuyo conocimiento nos permite establecer estrategias de prevención personalizada más eficaces.Obesity has been classified as the epidemic of the 21st century, whose prevalence worldwide has increased in recent decades. Obesity is defined as an abnormal or excessive accumulation of fat that can be harmful to health. Besides being considered a disease, it is a risk factor for other pathologies. The origin of obesity is multifactorial, and occurs as a result of the combination of genetic, environmental effects and their interactions. Environmental factors are modifiable factors that can be considered lifestyle, and genetic factors are considered non-modifiable, having an important role, between 40-80% in the susceptibility of the development of obesity. For this reason the objective of this doctoral thesis was to study the influence of environmental and genetic factors in obesity and related diseases in a population of obesity cases and controls of the OBENUTIC study(Obesity, Nutrition and Information and Communication Technologies) and in a population at cardiovascular risk, PREDIMED study (MEDITERRANEAN Diet Prevention). As well as analyzing the level of gene expression related to obesity and sleep after temporality in patients of general population. The most relevant results showed strong associations between lifestyle and obesity, being the performance of physical activity, sleep pattern and diet quality the most important factors in obesity and associated parameters. Regarding the genetic factors, it was observed that polymorphisms in taste receptors genes were related to the perception of certain flavors and to the consumption of characteristic foods. In turn, these genetic polymorphisms, such as rs713598 of the TAS2R38 gene and rs35874116 of the TAS1R2 gene showed association with parameters of obesity. On the other hand, genes associated with the circadian cycle (rs4580704 of the CLOCK gene and rs1982350 of the BMAL1 gene) showed association with BMI and plasma glucose level. Genetic variants of genes involved in dietary intake, lipid and carbohydrate metabolism such as rs9939606 of the FTO gene, rs3802177 of the gene SLC3048 or rs708272 of the CETP gene, were associated with obesity phenotypes. Furthermore, in patients in the PREDIMED study, polymorphisms in microRNAs (rs895819 in the pre-miR-27a, rs11614913 in miRNA196a2 and rs2292832 in miRNA-149) have shown an association with food consumption. Some of the polymorphisms analyzed presented environmental modulations related to obesity or associated parameters. On the other hand, changes in the expression level after temporality were observed and higher expression levels were observed in the FTO, CLOCK, PER1, CD36 and VRK2 genes in the individuals who had shown a significant weight loss. These results highlight the importance of the study of lifestyle factors, genetic factors and gene expression, as well as their interaction, whose knowledge allows us to establish more effective personalized prevention strategies

Metabolites, genomics, epigenomics, exposomics and health: Focus on serum bilirubin concentrations in subjects with metabolic syndrome from a Mediterranean population

Pòster presentat al congrés "Understanding Human Diseases Through Metabolomics: Interactions Among the Genome, Proteome, Gut Microbiome and Nutrition", Metabolomics and Human Health, Gordon Research Conference (February 3 - 8, 2019 Ventura, CA, United States)Although metabolomics aims at the measurement of small molecules (metabolites) in a biological sample, this knowledge requires additional information on the related genetic variants, epigenetic regulators and environmental factors (diet, smoking, physical activity, etc.) in order to translate the knowledge into actionable therapeutic or preventive evidence for complex disease outcomes. We focused on serum bilirubin, a metabolite generated when heme oxygenase catalyzes the degradation of heme (Figure 1). This produces biliverdin, which is converted into bilirubin by biliverdinreductase. Bilirubin is further processed in hepatocytes, where unconjugated bilirubin is conjugated by uridine diphosphate-glucuronosyltransferase (UDP-GT) to a water-soluble form for excretion. For decades, increased serum bilirubin concentrations were considered a threatening sign of underlying liver disease and had been associated with neonatal jaundice. However, data from recent years show that bilirubin is a powerful antioxidant and suggest that slightly increased serum bilirubin concentrations are protective against oxidative stress-related diseases

Nutrigenetics, nutrigenomics and Mediterranean diet: a new vision for gastronomy

Tanto la nutrigenética como la nutrigenómica son disciplinas dentro de la denominada genómica nutricional, que, en sentido amplio, proporciona el marco de integración de las distintas ómicas con las ciencias de la alimentación y nutrición. Tras décadas de estudios nutrigenéticos y nutrigenómicos, se dispone de una cantidad relevante de conocimientos para plantear su aplicación en la denominada nutrición de precisión. Esta nueva disciplina plantea que hay que tener en cuenta las características particulares de la persona para proporcionar la mejor dieta para prevenir o tratar la enfermedad. Los marcadores ómicos se consideran relevantes en dicha personalización. Existen muchos alimentos, nutrientes y patrones de dieta que se han investigado en nutrigenética y nutrigenómica; entre ellos, podemos mencionar el patrón de dieta mediterránea. A pesar de la heterogeneidad en la definición de dieta mediterránea, existen varios estudios que muestran que la dieta mediterránea puede interaccionar con el genoma, disminuyendo el riesgo de enfermedad en las personas genéticamente más susceptibles. Paralelamente, algunos estudios están mostrando los mecanismos por los que la dieta mediterránea puede ejercer este efecto protector. Conocer con más detalle la susceptibilidad genética, los mecanismos epigenéticos, la influencia del metaboloma y de otras ómicas puede ser relevante en gastronomía, entendida como la práctica del arte de elegir, cocinar y comer los alimentos. Esta influencia ómica no solo podemos encontrarla en los fenotipos de salud-enfermedad, sino también en la percepción del sabor y del olor de los alimentos (las preferencias por determinadas comidas). Todo ello, bien integrado, puede contribuir al incremento del disfrute a la vez que se sigue una alimentación saludable.Both nutrigenetics and nutrigenomics are disciplines that form part of what is known as Nutritional Genomics, which, in the widest sense, provides the framework for integrating different omics with food and nutrition sciences. After decades of nutrigenetic and nutrigenomic studies, there is a large enough amount of knowledge to consider its application in so-called precision nutrition. This new discipline seeks to take into account the particular characteristics of the individual in order to provide the best diet for preventing or treating a disease. Omic markers are considered to be of importance to that personalization. There are many foods, nutrients and dietary patterns that have been researched in nutrigenetics and nutrigenomics, including the Mediterranean Diet pattern. Despite heterogeneity in defining the Mediterranean Diet, there are various studies that show that the Mediterranean Diet can interact with the genome, so reducing the risk of disease in the most genetically susceptible individuals. Likewise, several studies have recently been revealing the mechanisms through which the Mediterranean Diet may exercise this protective effect. Understanding genetic susceptibility, epigenetic mechanisms, the influence of the metabolome and other omics in more detail may be important in gastronomy, understood as the practice of selecting, cooking and eating food. This omic influence can not only be found in health-disease phenotypes, but also in food taste and smell perception and preferences for certain dishes. Considering all of these together may contribute to an increase in enjoying and at the same time pursuing healthy eating

Crosstalk between smoking and the genome in older subjects with metabolic syndrome through genomics, epigenomics and transcriptomics

Pòster presentat a EMBO - EMBL Symposia Multiomics to Mechanisms - Challenges in Data Integration. September (11th – 13th 2019 European Molecular Biology Laboratory. Heidelberg, Germany)Tobacco smoking (Figure 1) is a major cause of cardiovascular diseases (CVD), and appears to have a multiplicative interaction with the other major CVD risk factors (lipids, hypertension, diabetes and others present in the metabolic syndrome (MetS). Several omics have analyzed the separate effects of tobacco smoking on the genome, epigenome, transcriptome, metabolome, etc. However an integrated omics approach can help to better understand the crosstalk between tobacco smoking and the genome

Genome-wide association analyses of weight loss in a randomized controlled trial of lifestyle intervention, and combined transcriptome-wide associations in a Mediterranean population

Pòster presentat a EMBO - EMBL Symposia Multiomics to Mechanisms - Challenges in Data Integration. September (11th – 13th 2019 European Molecular Biology Laboratory. Heidelberg, Germany)Although large-scale genome-wide association studies (GWAS) for obesity traits have identified more than 400 associated loci from observational studies (Figure 1), we highlight the fact that currently the number of GWAS for intentional weight change in randomized controlled trials (RCT) of lifestyle interventions is very scarce. Only a few RCT on weight loss have been carried out and recently a GWAS including 2 populations (a Canadian RCT and the Diogenes RCT) was been published (Valsesia et al, Nat Communications, 2019). Likely, at the transcriptome level, there is a scarcity of transcriptome-wide association studies (TWAS) of weight loss in RCT. Moreover, this scarcity is higher for studies including subjects from the Mediterranean countries. Therefore, our first aim was to undertake a GWAS in overweight/obese subjects from a Mediterranean population (Spain) after 1-year lifestyle intervention (including an energy restricted Mediterranean diet plus physical activity) in a RCT to identify genetic variants associated with weight loss and related outcomes. In addition, as a second aim, we carried out a TWAS in a subsample of subjects for the same intervention to identify changes in gene expression and related pathways

Circulating adiponectin and Its association with metabolic traits and Type 2 Diabetes: gene-diet interactions focusing on selected gene variants and at the genome-wide level in high-cardiovascular risk mediterranean subjects

Adiponectin is gaining renewed interest since, in addition to its possible protective role against insulin resistance and arteriosclerosis, recent studies suggest other additional favorable effects. However, the influence of gene-diet interactions on plasma adiponectin levels is still little understood. We analyzed the association between plasma adiponectin levels and various metabolic traits in a high-cardiovascular risk Mediterranean population, as well as the genetic effect of four candidate single-nucleotide polymorphisms (SNPs) in the adiponectin gene (ADIPOQ) and their interactions with the Mediterranean dietary pattern. Additionally, we explored, at the genome-wide level, the SNPs most associated with plasma adiponectin levels, as well as gene–diet interactions with the Mediterranean diet. In the 954 participants studied (aged 55–80 years), plasma adiponectin levels were strongly associated with plasma HDL-C concentrations (p = 6.6 × 10−36) and inversely related to triglycerides (p = 4.7 × 10−18), fasting glucose (p = 3.5 × 10−16) and type 2 diabetes (p = 1.4 × 10−7). Of the four pre-selected ADIPOQ candidate SNPs, the one most associated with plasma adiponectin was the −11391G > A (rs17300539) promoter SNP (p = 7.2 × 10−5, in the multivariable adjusted model). No significant interactions with the Mediterranean diet pattern were observed for these SNPs. Additionally, in the exploratory genome-wide association study (GWAS), we found new SNPs associated with adiponectin concentrations at the suggestive genome-wide level (p < 1 × 10−5) for the whole population, including the lead SNP rs9738548 (intergenic) and rs11647294 in the VAT1L (Vesicle Amine Transport 1 Like) gene. We also found other promising SNPs on exploring different strata such as men, women, diabetics and non-diabetics (p = 3.5 × 10−8 for rs2850066). Similarly, we explored gene–Mediterranean diet interactions at the GWAS level and identified several SNPs with gene–diet interactions at p < 1 × 10−5. A remarkable gene–diet interaction was revealed for the rs2917570 SNP in the OPCML (Opioid Binding Protein/Cell Adhesion Molecule Like) gene, previously reported to be associated with adiponectin levels in some populations. Our results suggest that, in this high-cardiovascular risk Mediterranean population, and even though adiponectin is favorably associated with metabolic traits and lower type 2 diabetes, the gene variants more associated with adiponectin may be population-specific, and some suggestive gene–Mediterranean diet interactions were detected

Influence of Demographic and Lifestyle Variables on Plasma Magnesium Concentrations and Their Associations with Cardiovascular Risk Factors in a Mediterranean Population

Several studies have shown that a low magnesium (Mg) intake in the diet is associated with greater cardiovascular risk and greater risk of diabetes. However, the results are not consistent in all populations. To minimize the biases derived from diet measurement, more objective biomarkers of magnesium status have been proposed. Although there is still no ideal biomarker for Mg, several studies have shown that plasma Mg concentrations could be a relatively acceptable biomarker for cardiovascular risk assessment. However, further studies are required to better characterize this marker in different populations. Our aim was to analyze the association between plasma Mg concentrations (measured through inductively coupled plasma mass spectrometry (ICP-MS)) methods, and cardiovascular risk factors in individuals from a general Mediterranean population (aged 18–80 years). The influence of demographic and lifestyle variables, including adherence to the Mediterranean diet, on plasma Mg concentrations was analyzed. The mean Mg level of the population studied was 0.77 ± 0.08 mmol/L, the prevalence of hypomagnesemia (<0.70 mmol/L) being 18.6%. We did not find any statistically significant differences between plasma Mg concentrations and sex, age, tobacco smoking and total adherence to the Mediterranean diet (p > 0.05). We found a statistically significant association between plasma Mg concentrations and the prevalence of type-2 diabetes (0.77 ± 0.08 mmol/L in non-diabetics versus 0.73 ± 0.13 mmol/L in diabetics; p = 0.009). Despite the low prevalence of type-2 diabetes in this population (11.24% in subjects with hypomagnesemia versus 3.91%, in normomagnesemia; p = 0.005), hypomagnesemia was associated with greater odds of being diabetic in comparison with normomagnesemia (OR = 3.36; p = 0.016, even after adjustment for sex, age, obesity, and medications). On the other hand, no statistically significant association of plasma Mg concentrations with obesity, hypertension, fasting triglycerides, HDL-cholesterol or uric acid was found. However, in contrast to what was initially expected, a statistically significant association was found between plasma Mg concentrations (basically in the highest quartile) and greater total cholesterol (p < 0.05) and LDL-cholesterol concentrations (p < 0.05). In conclusion, our results contribute to increasing the evidence gathered by numerous studies on the inverse association between hypomagnesemia and type-2 diabetes, as well as to the observation, previously reported in some studies, of a direct association with hypercholesterolemia. This paradoxical link should be deeply investigated in further studies.This study was partially funded, by the Spanish Ministry of Health (Instituto de Salud Carlos III) and the Ministerio de Economía y Competitividad-Fondo Europeo de Desarrollo Regional (FEDER) (grants CIBER 06/03, SAF2016–80532-R); the Junta de Andalucía (AGR145 research group); the University Jaume I (grant P1–1B2013–54); the Fundació La Marató de TV3 (grant 538/U/2016) and the Generalitat Valenciana (grants PROMETEO2017/017, and APOSTD/2019/136)

Influence of DNA-Polymorphisms in Selected Circadian Clock Genes on Clock Gene Expression in Subjects from the General Population and Their Association with Sleep Duration

Background and Objectives: Circadian rhythms have an important implication in numerous physiological and metabolic processes, including the sleep/wake cycle. Inter-individual differences in factors associated with circadian system may be due to gene differences in gene expression. Although several studies have analyzed the association between DNA polymorphisms and circadian variables, the influence on gene expression has been poorly analyzed. Our goal was to analyze the association of genetic variations in the clock genes and the gene expression level. Materials and Methods: We carried out a cross-sectional study of 102 adults (50.9% women). RNA and DNA were isolated from blood and single-nucleotide polymorphisms (SNPs), and the main circadian clock genes were determined. Gene expression of CLOCK, PER1, and VRK2 genes was measured by Reverse-transcription polymerase chain reaction (RT-PCR). The association between the DNA-SNPs and gene expression was analyzed at the gene level. In addition, a polygenic risk score (PRS), including all the significant SNPs related to gene expression, was created for each gene. Multivariable model analysis was performed. Results: Sex-specific differences were detected in PER1 expression, with these being higher in women (p = 0.034). No significant differences were detected in clock genes expression and lifestyle variables. We observed a significant association between the ARNTL-rs7924734, ARNTL-rs10832027, VRK2- rs2678902 SNPs, and CLOCK gene expression; the PER3-rs228642 and PER3-rs10127838 were related to PER1 expression, and the ARNTL-rs10832027, ARNTL-rs11022778, and MNTR1B-rs10830963 were associated with VRK2 gene expression (p < 0.05). The specific PRS created was significantly associated with each of the gene expressions analyzed (p < 0.001). Finally, sleep duration was associated with PER3-rs238666 (p = 0.008) and CLOCK-rs4580704 (p = 0.023). Conclusion: We detected significant associations between DNA-SNPs in the clock genes and their gene expression level in leukocytes and observed some differences in gene expression per sex. Moreover, we reported for the first time an association between clock gene polymorphisms and CLOCK, PER1, and VRK2 gene expression. These findings need further investigation

Chronological age interacts with the circadian melatonin receptor 1b gene variation, determining fasting glucose concentrations in mediterranean populations. Additional analyses on type-2 diabetes risk

Gene-age interactions have not been systematically investigated on metabolic phenotypes and this modulation will be key for a better understanding of the temporal regulation in nutrigenomics. Taking into account that aging is typically associated with both impairment of the circadian system and a decrease in melatonin secretion, we focused on the melatonin receptor 1B (MTNR1B)-rs10830963 C>G variant that has been associated with fasting glucose concentrations, gestational diabetes, and type-2 diabetes. Therefore, our main aim was to investigate whether the association between the MTNR1B-rs10830963 polymorphism and fasting glucose is age dependent. Our secondary aims were to analyze the polymorphism association with type-2 diabetes and explore the gene-pregnancies interactions on the later type-2 diabetes risk. Three Mediterranean cohorts (n = 2823) were analyzed. First, a cross-sectional study in the discovery cohort consisting of 1378 participants (aged 18 to 80 years; mean age 41 years) from the general population was carried out. To validate and extend the results, two replication cohorts consisting of elderly individuals were studied. In the discovery cohort, we observed a strong gene-age interaction (p = 0.001), determining fasting glucose in such a way that the increasing effect of the risk G-allele was much greater in young (p = 5.9 × 10−10) than in elderly participants (p = 0.805). Consistently, the association of the MTNR1B-rs10830963 polymorphism with fasting glucose concentrations in the two replication cohorts (mean age over 65 years) did not reach statistical significance (p > 0.05 for both). However, in the elderly cohorts, significant associations between the polymorphism and type-2 diabetes at baseline were found. Moreover, in one of the cohorts, we obtained a statistically significant interaction between the MTNR1B polymorphism and the number of pregnancies, retrospectively assessed, on the type-2 diabetes risk. In conclusion, the association of the MTNR1B-rs10830963 polymorphism with fasting glucose is age-dependent, having a greater effect in younger people. However, in elderly subjects, associations of the polymorphism with type-2 diabetes were observed and our exploratory analysis suggested a modulatory effect of the number of past pregnancies on the future type-2 diabetes genetic risk

Genome-Wide Association Study for Serum Omega-3 and Omega-6 Polyunsaturated Fatty Acids: Exploratory Analysis of the Sex-Specific Effects and Dietary Modulation in Mediterranean Subjects with Metabolic Syndrome.

Many early studies presented beneficial effects of polyunsaturated fatty acids (PUFA) on cardiovascular risk factors and disease. However, results from recent meta-analyses indicate that this effect would be very low or nil. One of the factors that may contribute to the inconsistency of the results is that, in most studies, genetic factors have not been taken into consideration. It is known that fatty acid desaturase (FADS) gene cluster in chromosome 11 is a very important determinant of plasma PUFA, and that the prevalence of the single nucleotide polymorphisms (SNPs) varies greatly between populations and may constitute a bias in meta-analyses. Previous genome-wide association studies (GWAS) have been carried out in other populations and none of them have investigated sex and Mediterranean dietary pattern interactions at the genome-wide level. Our aims were to undertake a GWAS to discover the genes most associated with serum PUFA concentrations (omega-3, omega-6, and some fatty acids) in a scarcely studied Mediterranean population with metabolic syndrome, and to explore sex and adherence to Mediterranean diet (MedDiet) interactions at the genome-wide level. Serum PUFA were determined by NMR spectroscopy. We found strong robust associations between various SNPs in the FADS cluster and omega-3 concentrations (top-ranked in the adjusted model: FADS1-rs174547, p = 3.34 × 10-14; FADS1-rs174550, p = 5.35 × 10-14; FADS2-rs1535, p = 5.85 × 10-14; FADS1-rs174546, p = 6.72 × 10-14; FADS2-rs174546, p = 9.75 × 10-14; FADS2- rs174576, p = 1.17 × 10-13; FADS2-rs174577, p = 1.12 × 10-12, among others). We also detected a genome-wide significant association with other genes in chromosome 11: MYRF (myelin regulatory factor)-rs174535, p = 1.49 × 10-12; TMEM258 (transmembrane protein 258)-rs102275, p = 2.43 × 10-12; FEN1 (flap structure-specific endonuclease 1)-rs174538, p = 1.96 × 10-11). Similar genome-wide statistically significant results were found for docosahexaenoic fatty acid (DHA). However, no such associations were detected for omega-6 PUFAs or linoleic acid (LA). For total PUFA, we observed a consistent gene*sex interaction with the DNTTIP2 (deoxynucleotidyl transferase terminal interacting protein 2)-rs3747965 p = 1.36 × 10-8. For adherence to MedDiet, we obtained a relevant interaction with the ME1 (malic enzyme 1) gene (a gene strongly regulated by fat) in determining serum omega-3. The top-ranked SNP for this interaction was ME1-rs3798890 (p = 2.15 × 10-7). In the regional-wide association study, specifically focused on the FADS1/FASD2/FADS3 and ELOVL (fatty acid elongase) 2/ELOVL 5 regions, we detected several statistically significant associations at p < 0.05. In conclusion, our results confirm a robust role of the FADS cluster on serum PUFA in this population, but the associations vary depending on the PUFA. Moreover, the detection of some sex and diet interactions underlines the need for these associations/interactions to be studied in all specific populations so as to better understand the complex metabolism of PUFA.This study was partially funded, by the Spanish Ministry of Health (Instituto de Salud Carlos III) and the Ministerio de Economía y Competitividad-Fondo Europeo de Desarrollo Regional (FEDER) (grants CIBER 06/03, PI06/1326, PI13/00728, PI16/00366, SAF2016–80532-R and FPU 18/01703); the University Jaume I (grants P1–1B2013–54 and COGRUP/2016/06); the Rei Jaume I Award for Medical Research 2018; the Fundació La Marató de TV3 (grant 538/U/2016); and the Generalitat Valenciana (grants PROMETEO2017/017, AEST/2018/044 and APOSTD/2019/136) and the US Department of Agriculture, Agriculture Research Service (grant 8050–51000--098--00D).N