Consejería de Agricultura, Pesca, Agua y Desarrollo Rural

Wine is one of the most consumed beverages around the world. It is composed of alcohols, sugars, acids, minerals, proteins and other compounds, such as organic acids and volatile and phenolic compounds (also called polyphenols). Polyphenols have been shown to be highly related to both (i) wine quality (color, flavor, and taste) and (ii) health-promoting properties (antioxidant and cardioprotective among others). Polyphenols can be grouped into two big families: (i) Flavonoids, including anthocyanidins, flavonols, flavanols, hydrolysable and condensed tannins, flavanones, flavones and chalcones; and (ii) Non-flavonoids, including hydroxycinnamic acids, hydroxybenzoic acids, stilbenes, tyrosol and hydroxytyrosol. Each group affects in some way the different properties of wine to a greater or a lesser extent. For that reason, the phenolic composition can be managed to obtain singular wines with specific, desirable characteristics. The current review presents a summary of the ways in which the phenolic composition of wine can be modulated, including (a) invariable factors such as variety, field management or climatic conditions; (b) pre-fermentative strategies such as maceration, thermovinification and pulsed electric field; (c) fermentative strategies such as the use of different yeasts and bacteria; and (d) post-fermentative strategies such as maceration, fining agents and aging. Finally, the different extraction methods and analytical techniques used for polyphenol detection and quantification have been also reviewed.

Field surveys were conducted in the autumn of 1997 in the main tomato (Lycopersicon esculentum)-growing regions of southern Spain following a severe tomato yellow leaf curl epidemic in tomato. Tomato yellow leaf curl virus (TYLCV)-Is was found to have spread to all regions and to coexist with TYLCV-Sr, which has been present since 1992. TYLCV-Is was also shown to be the causal agent of bean leaf crumple, a novel disease that has caused severe economic losses in fresh-market common bean (Phaseolus vulgaris) crops of southern Spain since September 1997. The disease was reproduced by infecting beans with cloned TYLCV-Is obtained from infected tomato plants collected in Almería. This is the first report of bean leaf crumple disease and the first report of a geminivirus in bean from Spain.

To identify Cd- and Zn-accumulating plants exhibiting a high growth rate, seeds from the halophyte species Mediterranean saltbush (Atriplex halimus L.) were collected on a heavy-metal-contaminated site in southeastern Spain (Llano del Beal, Cartagena). Seedlings from this ecotype were exposed for 3 wk to 0.1 mM Cd or Zn in a nutrient solution in a fully controlled environment. All plants remained alive and no significant growth inhibition was recorded until the end of the experiment. Mean Cd and Zn accumulation in aerial parts was 830 and 440 mg kg(-1), respectively, and the rate of metal translocation even increased with the duration of stress exposure. Resistance to heavy metals in this species may be partly linked to precipitation of Cd in oxalate crystals in the stems. A Cd-induced decrease in glutathione concentration also suggests that phytochelatins overproduction may occur in these conditions. We conclude that Mediterranean saltbush, which is able to produce up to 5 Mg dry matter ha(-1) yr(-1), may be an effective species for phytoextraction and should be tested for this purpose in field conditions.

Abstract. The impact of road dust emissions on PM10 and PM2.5 (atmospheric particulate matter with diameteer < 10 μm and 2.5 μm mass concentrations recorded from 2003 to 2010 at 11 locations (rural, urban and industrial) in southern Spain was estimated based on the chemical characterization of PM and the use of a constrained Positive Matrix Factorization, where the chemical profile of local road dust samples is used as a priori knowledge. Results indicate that road dust increased PM10 levels on average by 21–35% at traffic sites, 29–34% at urban background sites heavily affected by road traffic emissions, 17–22% at urban-industrial sites and 9–22% at rural sites. Road dust contributions to ambient PM levels show a marked seasonality with maxima in summer and minima in winter, likely due to the rainfall frequency. Decreasing concentration trends over the sampling years were found at some traffic and urban sites but in most cases the decreases were less significant than for vehicle exhaust emissions, while concentrations increased at industrial sites, probably due to local peculiarities. Concerning PM2.5, road dust contributions were lower than in PM10, as expected but still important (21–31%, 11–31%, 6–16% and 7% for traffic, urban background, urban-industrial and rural sites, respectively). In addition the three main sources of road dust (carbonaceous particles, brake wear and road wear/mineral) were identified and their contributions to road dust mass loadings estimated, supporting the idea that air quality managers should drive measures aimed at preventing the build-up of road dust particles on roads.

The increased frequency of harmful algal blooms (HABs) is a major environmental concern worldwide, resulting not only in increased treatment costs for drinking water but also in impacts on tourism, commercial fishing and aquaculture and risks to human and animal health. Traditional strategies with ship-based approaches based on field sampling and laboratory analysis have been adopted to assess HABs. However, these methods are labour intensive and costly and do not provide synoptic views of the bloom conditions. Here, we show that the Sentinel-2 twin satellite mission of the Copernicus programme, in combination with in situ data, is a powerful tool that can offer valuable spatiotemporal information about a bloom of the dinoflagellate Lingulodinium polyedra that occurred on the SW Iberian Peninsula. Using the robust ACOLITE atmospheric correction processor combined with the normalized difference chlorophyll index (NDCI), the enhanced mapping of small blooms can be performed at a 10 m spatial resolution, revealing surface patches and a heterogeneous distribution. This research also demonstrates the improved capabilities of Sentinel-2 compared to those of Landsat-8 and Sentinel-3 for continuous monitoring. The Sentinel-3 and Sentinel-2 missions provide ecosystem observations that allow the environmental community and water managers to evaluate changes in water quality and bloom distribution and that facilitate field-based measurements. Therefore, the value added by the Copernicus products in terms of frequency and synoptic observations is of paramount importance for ecological and management purposes at regional and national scales.

Current understanding of the postglacial colonization of Nearctic and Palearctic species relies heavily on inferences drawn from the phylogeographic analysis of contemporary generic variants. Modern postglacial populations are supposed to be representative of their Pleistocene ancestors, and their current distribution is assumed to reflect the different colonization success and dispersal patterns of refugial lineages. Yet, testing of phylogeographic models against ancestral genomes from glacial refugia has rarely been possible. Here we compare ND1 mitochondrial DNA variation in late Pleistocene (16,000-40,000 years before present), historical and contemporary Atlantic salmon (Salmo salar) populations from northern Spain and other regions of western Europe. Our study demonstrates the presence of Atlantic salmon in the Iberian glacial refugium during the last 40,000 years and points to the Iberian Peninsula as the likely source of the most common haplotype within the Atlantic lineage in Europe. However, our findings also suggest that there may have been significant changes in the genetic structure of the Iberian refugial stock since the last ice age, and question whether modern populations in refugial areas are representative of ice age populations. A common haplotype that persisted in the Iberian Peninsula during the Pleistocene last glacial maximum is now extremely rare or absent from European rivers, highlighting the need for caution when making phylogeographic inferences about the origin and distribution of modern genetic types.

Novel species of fungi described in this study include those from various countries as follows: Antartica , Cladosporium austrolitorale from coastal sea sand. Australia , Austroboletus yourkae on soil, Crepidotus innuopurpureus on dead wood, Curvularia stenotaphri from roots and leaves of Stenotaphrum secundatum and Thecaphora stajsicii from capsules of Oxalis radicosa. Belgium , Paraxerochrysium coryli (incl. Paraxerochrysium gen. nov.) from Corylus avellana. Brazil , Calvatia nordestina on soil, Didymella tabebuiicola from leaf spots on Tabebuia aurea, Fusarium subflagellisporum from hypertrophied floral and vegetative branches of Mangifera indica and Microdochium maculosum from living leaves of Digitaria insularis. Canada , Cuphophyllus bondii fromagrassland. Croatia , Mollisia inferiseptata from a rotten Laurus nobilis trunk. Cyprus , Amanita exilis oncalcareoussoil. Czech Republic , Cytospora hippophaicola from wood of symptomatic Vaccinium corymbosum. Denmark , Lasiosphaeria deviata on pieces of wood and herbaceousdebris. Dominican Republic , Calocybella goethei among grass on a lawn. France (Corsica) , Inocybe corsica onwetground. France (French Guiana) , Trechispora patawaensis on decayed branch of unknown angiosperm tree and Trechispora subregularis on decayed log of unknown angiosperm tree. Germany , Paramicrothecium sambuci (incl. Paramicrothecium gen. nov.)ondeadstemsof Sambucus nigra. India , Aureobasidium microtermitis from the gut of a Microtermes sp. termite, Laccaria diospyricola on soil and Phylloporia tamilnadensis on branches of Catunaregam spinosa . Iran , Pythium serotinoosporum from soil under Prunus dulcis. Italy , Pluteus brunneovenosus on twigs of broad leaved trees on the ground. Japan , Heterophoma rehmanniae on leaves of Rehmannia glutinosa f. hueichingensis. Kazakhstan , Murispora kazachstanica from healthy roots of Triticum aestivum. Namibia , Caespitomonium euphorbiae (incl. Caespitomonium gen. nov.)from stems of an Euphorbia sp. Netherlands , Alfaria junci, Myrmecridium junci, Myrmecridium juncicola, Myrmecridium juncigenum, Ophioceras junci, Paradinemasporium junci (incl. Paradinemasporium gen. nov.), Phialoseptomonium junci, Sporidesmiella juncicola, Xenopyricularia junci and Zaanenomyces quadripartis (incl. Zaanenomyces gen. nov.), fromdeadculmsof Juncus effusus, Cylindromonium everniae and Rhodoveronaea everniae from Evernia prunastri, Cyphellophora sambuci and Myrmecridium sambuci from Sambucus nigra, Kiflimonium junci, Saro cladium junci, Zaanenomyces moderatricis academiae and Zaanenomyces versatilis from dead culms of Juncus inflexus, Microcera physciae from Physcia tenella, Myrmecridium dactylidis from dead culms of Dactylis glomerata, Neochalara spiraeae and Sporidesmium spiraeae from leaves of Spiraea japonica, Neofabraea salicina from Salix sp., Paradissoconium narthecii (incl. Paradissoconium gen. nov.)from dead leaves of Narthecium ossifragum, Polyscytalum vaccinii from Vaccinium myrtillus, Pseudosoloacrosporiella cryptomeriae (incl. Pseudosoloacrosporiella gen. nov.)fromleavesof Cryptomeria japonica, Ramularia pararhabdospora from Plantago lanceolata, Sporidesmiella pini from needles of Pinus sylvestris and Xenoacrodontium juglandis (incl. Xenoacrodontium gen. nov. and Xenoacrodontiaceae fam. nov.)from Juglans regia . New Zealand , Cryptometrion metrosideri from twigs of Metrosideros sp., Coccomyces pycnophyllocladi from dead leaves of Phyllocladus alpinus, Hypoderma aliforme from fallen leaves Fuscopora solandri and Hypoderma subiculatum from dead leaves Phormium tenax. Norway , Neodevriesia kalakoutskii from permafrost and Variabilispora viridis from driftwood of Picea abies. Portugal , Entomortierella hereditatis from abio film covering adeteriorated limestone wall. Russia , Colpoma junipericola from needles of Juniperus sabina, Entoloma cinnamomeum on soil in grasslands, Entoloma verae on soil in grasslands, Hyphodermella pallidostraminea on a dry dead branch of Actinidia sp., Lepiota sayanensis onlitterinamixedforest, Papiliotrema horticola from Malus communis , Paramacroventuria ribis (incl. Paramacroventuria gen. nov.)fromleaves of Ribes aureum and Paramyrothecium lathyri from leaves of Lathyrus tuberosus. South Africa , Harzia combreti from leaf litter of Combretum collinum ssp. sulvense, Penicillium xyleborini from Xyleborinus saxesenii , Phaeoisaria dalbergiae from bark of Dalbergia armata, Protocreopsis euphorbiae from leaf litter of Euphorbia ingens and Roigiella syzygii from twigs of Syzygium chordatum . Spain , Genea zamorana on sandy soil, Gymnopus nigrescens on Scleropodium touretii, Hesperomyces parexochomi on Parexochomus quadriplagiatus, Paraphoma variabilis from dung, Phaeococcomyces kinklidomatophilus from a blackened metal railing of an industrial warehouse and Tuber suaveolens in soil under Quercus faginea. Svalbard and Jan Mayen , Inocybe nivea associated with Salix polaris. Thailand , Biscogniauxia whalleyi oncorticatedwood. UK , Parasitella quercicola from Quercus robur. USA , Aspergillus arizonicus from indoor air in a hospital, Caeliomyces tampanus (incl. Caeliomyces gen. nov.)fromoffice dust, Cippumomyces mortalis (incl. Cippumomyces gen. nov.)fromatombstone, Cylindrium desperesense from air in a store, Tetracoccosporium pseudoaerium from air sample in house, Toxicocladosporium glendoranum from air in a brick room, Toxicocladosporium losalamitosense from air in a classroom, Valsonectria portsmouthensis from airinmen'slockerroomand Varicosporellopsis americana from sludge in a water reservoir. Vietnam , Entoloma kovalenkoi on rotten wood, Fusarium chuoi </jats:

This paper reports the results on the agronomic performance of organic amendments in the EU 7th FP project “FERTIPLUS—reducing mineral fertilizers and agro-chemicals by recycling treated organic waste as compost and bio-char”. Four case studies on field-scale application of biochar, compost and biochar-blended compost were established and studied for three consecutive years in four distinct cropping systems and under different agro-climatic conditions in Europe. These included the following sites: olive groves in Murcia (Spain), greenhouse grown tomatoes in Almeria (Spain), an arable crop rotation in Oost-Vlaanderen (Merelbeke, Belgium), and three vineyards in Friuli Venezia Giulia (Italy). A slow pyrolysis oak biochar was applied, either alone or in combination with organic residues: compost from olive wastes in Murcia (Spain), sheep manure in Almeria (Spain), and compost from biowaste and green waste in Belgium and Italy. The agronomical benefits were evaluated based on different aspects of soil fertility (soil total organic carbon (TOC), pH, nutrient cycling and microbial activity) and crop nutritional status and productivity. All amendments were effective in increasing soil organic C in all the field trials. On average, the increase with respect to the control was about 11% for compost, 20% for biochar-blended compost, and 36% for biochar. The amendments also raised the pH by 0.15–0.50 units in acidic soils. Only biochar had a negligible fertilization effect. On the contrary, compost and biochar-blended compost were effective in enhancing soil fertility by increasing nutrient cycling (25% mean increase in extractable organic C and 44% increase in extractable N), element availability (26% increase in available K), and soil microbial activity (26% increase in soil respiration and 2–4 fold enhancement of denitrifying activity). In general, the tested amendments did not show any negative effect on crop yield and quality. Furthermore, in vineyards and greenhouse grown tomatoes cropping systems, compost and biochar-blended compost were also effective in enhancing key crop quality parameters (9% increase in grape must acidity and 16% increase in weight, 9% increase in diameter and 8% increase in hardness of tomato fruits) important for the quality and marketability of the crops. The overall results of the project suggest that the application of a mixture of biochar and compost can benefit crops. Therefore, biochar-blended compost can support and maintain soil fertility.

This paper describes the first isolations of African horse sickness virus (AHSV) from insects in Spain. Seven isolations of AHSV serotype 4 were made; four from Culicoides imicola a known vector of the virus elsewhere, two from mixed pools of Culicoides species not including C. imicola and one from blood engorged mosquitoes. Three further isolations of AHSV serotype 4 were also made from horses kept adjacent to the insect collecting sites. This work presents the first definitive identification of the vectors of AHSV in Spain during the 1987, 88 and 89 epizootics. Suggestions are also made concerning the significance of these findings with regard to the epidemiology of African horse sickness in Spain.

In September 2012, a novel disease syndrome was observed in zucchini (Cucurbita pepo L.) crops in Murcia Province (southeastern Spain). Symptoms included curling, vein swelling, and severe mosaic in young leaves, short internodes, and fruit skin roughness, resembling begomovirus infection. Similar symptoms were observed in May 2013 in Almería Province (southern Spain). DNA was isolated from 8 and 7 symptomatic leaf samples collected in Murcia and Almería, respectively, and analyzed by PCR with primers GemCP-V-5' and GemCP-C-3' designed to detect begomoviruses by amplifying the core of coat protein gene (CP) (3). DNA fragments of the expected size (~600 bp) were amplified supporting a begomovirus infection. The DNA sequences obtained from four samples were identical. BLAST analysis showed the highest nucleotide identity (98%) with partial CP gene sequences from isolates of Tomato leaf curl New Delhi virus (ToLCNDV) infecting cucumber in India (GenBank Accession No. KC846817). ToLCNDV, a bipartite begomovirus first reported from tomato, also infects other solanaceous and cucurbitaceous crops in India and neighboring countries (1). DNA from two samples from Murcia and three from Almería was used for rolling-circle amplification using ϕ29 DNA polymerase (TempliPhi kit, GE Healthcare, Little Chalfont, UK) and digested with a set of restriction endonucleases. All five samples yielded amplification products with identical restriction patterns. Two samples from Murcia (MU-8.1 and MU-11.1) and one from Almería (AL-661) were selected to clone the putative DNA-A and DNA-B begomovirus genome components by using single BamHI or NcoI sites. Inserts of two clones from each sample, one corresponding to DNA-A and one to DNA-B, were completely sequenced. The cloned genomes exhibited the typical organization of Old World bipartite begomoviruses (1). Sequences were aligned with begomovirus sequences available in databases using MUSCLE and pairwise identity scores were calculated with SDT (species demarcation tool [4]). DNA-A sequences obtained from Murcia (2,738 nt, KF749224 and KF749225) and Almería (2,738 nt, KF749223) shared >99% nucleotide identity, with the highest nucleotide identity (91.3 to 91.5%) with that of an Indian ToLCNDV isolate from chilli (HM007120). DNA-B sequences (2,684 nt, KF749226, KF749227, and KF749228) shared >99% nucleotide identity, and showed the highest nucleotide identity (83.1 to 83.3%) with that of a Pakistani ToLCNDV isolate from Solanum nigrum (AJ620188). Nucleotide sequence identity of DNA-A with the most closely related begomoviruses was above the 91% threshold for species demarcation (2), thus confirming that the begomoviruses found infecting zucchini in Spain are isolates of ToLCNDV. In fall 2013, the disease was widespread in zucchini both in Murcia and Almería, and ToLCNDV has also been found infecting melon and cucumber crops. To our knowledge, this is the first report of a bipartite begomovirus in Spain and Europe. References: (1) J. K. Brown et al. Page 351 in: Virus Taxonomy. Ninth Report of the ICTV. A. M. Q. King et al., eds. Elsevier/Academic Press, London, 2012. (2) ICTV Geminiviridae Study Group. New species and revised taxonomy proposal for the genus Begomovirus (Geminiviridae). ICTV. Retrieved from http://talk.ictvonline.org/files/proposals/ taxonomy_proposals_plant1/m/plant04/4720.aspx , 10 October 2013. (3) H. Lecoq and C. Desbiez. Adv. Virus Res. 84:67, 2012. (4) B. Muhire et al. Arch. Virol. 158:1411, 2013.

Gray mold, caused by the necrotrophic fungus Botrytis cinerea., is one of the most economically important diseases of strawberry. Gray mold control involves the application of fungicides throughout the strawberry growing season; however, B. cinerea isolates resistant to multiple classes of site-specific fungicides have been recently reported in the Spanish gray mold population. Succinate dehydrogenase inhibitors (SDHI) constitute a relatively novel class of fungicides registered for gray mold control representing new alternatives for strawberry growers. In the present study, 37 B. cinerea isolates previously characterized for their sensitivity to boscalid and amino acid changes in the SdhB protein were used to determine the effective concentration that reduces mycelial growth by 50% (EC 50 ) to fluopyram, fluxapyroxad, and penthiopyrad. The present study was also conducted to obtain discriminatory doses to monitor SDHI fungicide resistance in 580 B. cinerea isolates collected from 27 commercial fields in Spain during 2014, 2015, and 2016. The EC 50 values ranged from 0.01 to &gt;100 μg/ml for fluopyram, &lt;0.01 to 4.19 μg/ml for fluxapyroxad, and, finally, &lt;0.01 to 59.65 μg/ml for penthiopyrad. Based on these results, as well as findings from a previous publication, the discriminatory doses chosen to examine sensitivities to boscalid, fluopyram, fluxapyroxad, and penthiopyrad were 100, 15, 1, and 6 μg/ml, respectively. Over the course of the 3-year monitoring period, the overall frequencies of resistance to the four SDHI were 56.9, 6.9, 12.9, and 24.6%, respectively. The frequency of boscalid-resistant isolates decreased from 73 to 41% over the years; however, the fluopyram-resistant isolates increased from 5 to 10% after 1 year of registration. Four SDHI resistance patterns were observed in our population, which included patterns I (30%; resistance to boscalid), II (13.8%; resistance to boscalid and penthiopyrad), III (5.7%; boscalid, fluxapyroxad, and penthiopyrad), and IV (7.9%; resistance to boscalid, fluopyram, fluxapyroxad, and penthiopyrad). Patterns I and II were associated with the amino acid substitutions H272R and H272Y; pattern III was associated only with the H272Y mutation; and, finally, pattern IV was associated with the N230I allele in the SdhB subunit. For gray mold management, it is suggested that the simultaneous use of boscalid and penthiopyrad should be limited to one application per season; however, fluxapyroxad and, especially, fluopyram could be used as valid SDHI alternatives for gray mold control, although they should be applied with caution.

Genetic evidence for the selective exploitation by anglers of early running fish was examined in four Iberian Atlantic salmon populations using protein and mtDNA markers. The populations studied had been exploited exclusively by anglers since 1949 during a fixed fishing season that ran approximately from March to July. Genetic variation at six protein loci was small and was accounted for by the MEP‐2 * and MDH‐3,4 * polymorphisms, which generally remained stable over time and were in Castle–Hardy–Weinberg equilibrium during the fishing season. Early running fish that had spent multiple winters at sea (MSW) generally had higher frequencies of the common MEP‐2 *(100) allele than did late running, one sea winter (1SW) grilse that were significantly smaller and tended to escape the fishery. Spawners differed from angler caught fish in their mtDNA frequencies and consistently had a lower sea‐age and a smaller body size. Spawners also smolted at an older age and displayed lower frequencies of the MEP‐2 *(100) allele in three of the four populations studied. These results suggest that in these rivers anglers selectively exploit a distinct component of the population and inadvertently cause a differential mortality of genetic types that is likely to be detrimental to population viability.

The essential oil of Chrysanthemum coronarium flowerheads showed strong nematicidal activity in vitro and in growth‐chamber experiments. Essential oil concentrations of 2, 4, 8 and 16 µ L mL −1 , significantly reduced hatch, J 2 survival (determined by final value and area under curves of cumulative percentage hatch or mortality) and reproduction rate of Meloidogyne artiellia in vitro , with the lowest values occurring at 16 µ L mL −1 . In pot trials with chickpea cv. PV 61, essential oil concentrations of 10–40 µ L per 500 cm 3 soil, applied on sterile cotton pellets, also significantly reduced the nematode's reproduction rate. The biological processes of mortality and hatching/reproduction were adequately described by the monomolecular and expanded negative exponential models, respectively. Effectiveness of soil amendment with either flowers, leaves, roots or seeds of C. coronarium , and flowers from several species of Asteraceae ( Chrysanthemum segetum , Calendula maritima , Calendula officinalis and Calendula suffruticosa ) at 5 g per 500 cm 3 soil was tested for suppression of M. artiellia and growth of chickpea cv. PV 61 under growth‐chamber conditions. In these tests, flowers of all five Asteraceae species and various parts of C. coronarium significantly reduced reproduction rates of M. artiellia , by 83·0–95·9%, with the minimum rates occurring in infected chickpea plants amended with flowers of C. officinalis and C. suffruticosa . The in vitro and in planta results suggest that the essential oil of C. coronarium and organic amendments from Asteraceae species may serve as nematicides.

La expansión del regadío a lo largo de las últimas décadas, y en los últimos quince años su modernización, ha necesitado incorporar gran cantidad de energía para la captación, transporte y distribución del agua a los cultivos. Esta transformación ha convertido al sector del regadío, que ocupa el 20% de la superficie cultivada en España, en el preponderante con casi el 60% de la producción final agraria. Al mismo tiempo se está avanzando rápidamente en la mejora de la eficiencia en el uso del agua, espoleado el sector por la falta de recursos suficientes en los años secos. Pero esta reducción unitaria de agua usada, del orden de un 21% desde 1950, debe ponerse en relación con la mayor intensidad de energía aportada al agua de riego que ha crecido desde 1950 en un 1800 %: la mayor eficiencia hídrica va ligada a una mayor ineficiencia energética. Los costes de la energía aportada al regadío crecen rápidamente año tras año, y pueden ser limitativos ante procesos de encarecimiento brusco del precio de la energía, especialmente el petróleo que sigue teniendo un gran peso en el mix energético español. Este riesgo se pondrá de manifiesto en los regadíos poco intensivos que utilizan aguas subterráneas o que se han implantado a cotas elevadas respecto a los puntos de captación. Los nuevos enfoques del uso de los recursos naturales a que nos obliga el cambio climático, y de manera más amplia la sostenibilidad, aconsejan introducir los balances energéticos del regadío, la huella hídrica y energética y los conceptos de agua y energía virtual, como maneras de acercarnos a las tendencias que deberán marcar los cambios necesarios para adaptar los regadíos a la sostenibilidad.

For centuries, domestic horses have represented an important means of transport and served as working and companion animals. Although their role in transportation is less important today, many horse breeds are still subject to intense selection based on their pattern of locomotion. A striking example of such a selected trait is the ability of a horse to perform additional gaits other than the common walk, trot and gallop. Those could be four-beat ambling gaits, which are particularly smooth and comfortable for the rider, or pace, used mainly in racing. Gaited horse breeds occur around the globe, suggesting that gaitedness is an old trait, selected for in many breeds. A recent study discovered that a nonsense mutation in DMRT3 has a major impact on gaitedness in horses and is present at a high frequency in gaited breeds and in horses bred for harness racing. Here, we report a study of the worldwide distribution of this mutation. We genotyped 4396 horses representing 141 horse breeds for the DMRT3 stop mutation. More than half (2749) of these horses also were genotyped for a SNP situated 32 kb upstream of the DMRT3 nonsense mutation because these two SNPs are in very strong linkage disequilibrium. We show that the DMRT3 mutation is present in 68 of the 141 genotyped horse breeds at a frequency ranging from 1% to 100%. We also show that the mutation is not limited to a geographical area, but is found worldwide. The breeds with a high frequency of the stop mutation (>50%) are either classified as gaited or bred for harness racing.

BACKGROUND: The main goal of the current work was to infer the demographic history of seven Spanish goat breeds (Malagueña, Murciano-Granadina, Florida, Palmera, Mallorquina, Bermeya and Blanca de Rasquera) based on genome-wide diversity data generated with the Illumina Goat SNP50 BeadChip (population size, N = 176). Five additional populations from Europe (Saanen and Carpathian) and Africa (Tunisian, Djallonké and Sahel) were also included in this analysis (N = 80) for comparative purposes. RESULTS: Our results show that the genetic background of Spanish goats traces back mainly to European breeds although signs of North African admixture were detected in two Andalusian breeds (Malagueña and Murciano-Granadina). In general, observed and expected heterozygosities were quite similar across the seven Spanish goat breeds under analysis irrespective of their population size and conservation status. For the Mallorquina and Blanca de Rasquera breeds, which have suffered strong population declines during the past decades, we observed increased frequencies of large-sized (ROH), a finding that is consistent with recent inbreeding. In contrast, a substantial part of the genome of the Palmera goat breed comprised short ROH, which suggests a strong and ancient founder effect. CONCLUSIONS: Admixture with African goats, genetic drift and inbreeding have had different effects across the seven Spanish goat breeds analysed in the current work. This has generated distinct patterns of genome-wide diversity that provide new clues about the demographic history of these populations.

The intradermal tuberculin (IDTB) test and the interferon-gamma (IFN-gamma) assay are used worldwide for detection of bovine tuberculosis in cattle, but little is known about the effect of co-infecting agents on the performance of these diagnostic tests. This report describes a field trial conducted in a cattle herd with dual infection (bovine tuberculosis and paratuberculosis) during 3.5 years. It has been based on a strategic approach encompassing serial parallel testing (comparative IDTB test, the IFN-gamma assay and serology of paratuberculosis) that was repeated 8 times over the period, and segregation of animals into two herds. The IDTB test detected 65.2% and the IFN-gamma test detected 69.6% of the Mycobacterium bovis culture-positive cattle. However, the IDTB test performed better during the first part of the trial, while the IFN-gamma test was the only method that detected infected animals during the following three samplings. The number of false positive reactors with the IDTB and/or the IFN-gamma tests was remarkably high compared to other reports, and could be caused by cross-reactivity with M. avium subsp. paratuberculosis. Also, the M. bovis isolates from cattle and wildlife from the same property were characterised using molecular techniques to disclose an epidemiological link. The IDTB test may not be appropriate to eradicate bovine tuberculosis in herds with dual mycobacterial infections. This report highlights the need to use several diagnostic techniques for the accurate detection of M. bovis infected animals in these herds.

Xylella fastidiosa, a quarantine organism in the European Union (EU), causes diseases in a wide variety of plants such as almond, cherry, grape, citrus, elm, olive, and coffee trees and many ornamental plants. Since the detection of the bacterium in Italy (2013), where it is associated to a severe epidemic on olive trees, the pathogen has also been detected in France (2015) and Germany (2016) (EPPO 2016). Due to the recent outbreaks and to different interceptions, the EU has implemented annual surveys in its member states to prevent new introductions or the spread of this harmful organism. During official surveys in late autumn 2016 in Mallorca Island, Spain, some cherry and Polygala myrtifolia plants located in a garden center near the locality of Manacor showed symptoms of marginal leaf scorch, leaf chlorosis, defoliation, and general decay. DNA was extracted from leaf veins and petioles of symptomatic leaves using the CTAB extraction method (EPPO 2016). DNA extracts were tested for the presence of X. fastidiosa by real-time PCR using two species-specific protocols (EPPO 2016): primers XF-F/XF-R and a dual-labeled probe XF-P (Harper et al. 2010, erratum 2013) and primers HL5/HL6 and Taqman probe (Francis et al. 2008). DNA from X. fastidiosa subsp. pauca strain CoDiRo or subsp. fastidiosa strain Temecula were used as positive controls. Three out of four cherry trees and four out of seven P. myrtifolia plants showing symptoms gave positive real-time amplification. These preliminary results were further confirmed by conventional PCR assays, using species specific primers RST31/RST33 (Minsavage et al. 1994), which yielded the expected amplicon of ∼700 bp of the RNA polymerase sigma 70 factor. Attempts to culture isolates from petioles tissues on PD2 and BCYE media failed for the P. myrtifolia plants. On the contrary, bacterial colonies typical of X. fastidiosa were recovered from one cherry plant after plate incubation for 14 days at 28°C. Colonies were suspended in water and boiled 10 min to obtain DNA before PCR analysis. Sequence analysis of the amplicon of the sigma 70 factor from all positive samples showed 95 to 100% identity with other X. fastidiosa sequences from GenBank, which confirmed the presence of X. fastidiosa DNA in the plants. Additionally, seven housekeeping genes (Yuan et al. 2010) were amplified using an annealing temperature of 58°C from both extracted plant or bacterial DNAs to identify the subspecies and sequence type (ST) of X. fastidiosa. Based on this MLST analysis, the ST1, related to X. fastidiosa subsp. fastidiosa, was found on one cherry tree and three P. myrtifolia plants. These four plants shared 100% nucleotide sequence identity for the sigma 70 factor (MF401540). Surprisingly, the fourth P. myrtifolia sample presented sequences that simultaneously have been only described in X. fastidiosa subsp. multiplex isolates (i.e., leuA_3, holC_3, nuoL_3, and gltT_3), although only four alleles could be amplified. This plant sample showed a sigma 70 factor sequence (MF401541) identical to that of X. fastidiosa subsp. multiplex isolates present in GenBank and different to those of samples infected by X. fastidiosa subsp. fastidiosa ST1. To our knowledge, this is the first detection of X. fastidiosa in Spain and of the subsp. fastidiosa on cherry and P. myrtifolia in Europe, underlining the presence of different genotypes in this island and the risk of introducing additional genetic diversity for X. fastidiosa to Europe. Eradication measures have been taken in the garden center according to the Spanish contingency plan and EU legislation.

Thirty-nine field populations of Frankliniella occidentalis (Pergande) were collected from different crops (sweet pepper, tomato, lettuce, artichoke, melon, cucumber, carnation, broad bean, peach and plum) in Murcia (south-east Spain). All populations were reared separately in the laboratory to obtain enough individuals for bioassays. Female thrips were bioassayed, using a standard topical application method, against methiocarb, methamidophos, acrinathrin, endosulfan, deltamethrin and formetanate. Methiocarb was the only insecticide that showed a high efficacy against F occidentalis at field dose rates. Acrinathrin and methamidophos were moderately effective, while endosulfan and deltamethrin were ineffective. Only moderate levels of resistance (Resistance Ratios at LC50 of 10-30) were detected for the selective insecticides methiocarb, formetanate and acrinathrin used against F occidentalis in crops where these insecticides are used intensively. This generalized and low level of resistance to these insecticides, coupled with a lack of efficacy for the three broad-spectrum insecticides, was observed even in intensively managed vegetable crops. Implementation of IPM strategies in Murcia has contributed to more successful insecticide anti-resistance management.

BACKGROUND: Senegalese sole (Solea senegalensis) and common sole (S. solea) are two economically and evolutionary important flatfish species both in fisheries and aquaculture. Although some genomic resources and tools were recently described in these species, further sequencing efforts are required to establish a complete transcriptome, and to identify new molecular markers. Moreover, the comparative analysis of transcriptomes will be useful to understand flatfish evolution. RESULTS: A comprehensive characterization of the transcriptome for each species was carried out using a large set of Illumina data (more than 1,800 millions reads for each sole species) and 454 reads (more than 5 millions reads only in S. senegalensis), providing coverages ranging from 1,384x to 2,543x. After a de novo assembly, 45,063 and 38,402 different transcripts were obtained, comprising 18,738 and 22,683 full-length cDNAs in S. senegalensis and S. solea, respectively. A reference transcriptome with the longest unique transcripts and putative non-redundant new transcripts was established for each species. A subset of 11,953 reference transcripts was qualified as highly reliable orthologs (>97% identity) between both species. A small subset of putative species-specific, lineage-specific and flatfish-specific transcripts were also identified. Furthermore, transcriptome data permitted the identification of single nucleotide polymorphisms and simple-sequence repeats confirmed by FISH to be used in further genetic and expression studies. Moreover, evidences on the retention of crystallins crybb1, crybb1-like and crybb3 in the two species of soles are also presented. Transcriptome information was applied to the design of a microarray tool in S. senegalensis that was successfully tested and validated by qPCR. Finally, transcriptomic data were hosted and structured at SoleaDB. CONCLUSIONS: Transcriptomes and molecular markers identified in this study represent a valuable source for future genomic studies in these economically important species. Orthology analysis provided new clues regarding sole genome evolution indicating a divergent evolution of crystallins in flatfish. The design of a microarray and establishment of a reference transcriptome will be useful for large-scale gene expression studies. Moreover, the integration of transcriptomic data in the SoleaDB will facilitate the management of genomic information in these important species.