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    An integrative "omics" approach identifies new candidate genes to impact aroma volatiles in peach fruit

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    Background: Ever since the recent completion of the peach genome, the focus of genetic research in this area has turned to the identification of genes related to important traits, such as fruit aroma volatiles. Of the over 100 volatile compounds described in peach, lactones most likely have the strongest effect on fruit aroma, while esters, terpenoids, and aldehydes have minor, yet significant effects. The identification of key genes underlying the production of aroma compounds is of interest for any fruit-quality improvement strategy. Results: Volatile (52 compounds) and gene expression (4348 genes) levels were profiled in peach fruit from a maturity time-course series belonging to two peach genotypes that showed considerable differences in maturation characteristics and postharvest ripening. This data set was analyzed by complementary correlation-based approaches to discover the genes related to the main aroma-contributing compounds: lactones, esters, and phenolic volatiles, among others. As a case study, one of the candidate genes was cloned and expressed in yeast to show specificity as an omega-6 Oleate desaturase, which may be involved in the production of a precursor of lactones/esters. Conclusions: Our approach revealed a set of genes (an alcohol acyl transferase, fatty acid desaturases, transcription factors, protein kinases, cytochromes, etc.) that are highly associated with peach fruit volatiles, and which could prove useful in breeding or for biotechnological purposes

    Genetic engineering of Plum pox virus resistance: 'HoneySweet' plum-from concept to product

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    Sharka disease, caused by Plum pox virus (PPV) was first recorded in Bulgaria during the early twentieth century and since that first report, the disease has progressively spread throughout Europe and more recently to Asia, Africa, North and South America. Few PPV resistance genes have been found to naturally occur in Prunus and this has led to the investigation of biotech approaches to the development of resistance through genetic engineering (GE). A notable example of the utility of this approach is ‘HoneySweet’ plum. PPV protection in this case is based on RNA interference (RNAi) and resistance has been shown to be highly effective, stable, durable, and heritable as a dominant trait. Extensive testing and risk assessment of ‘HoneySweet’ in laboratory, greenhouse and in the field for over 20 years has demonstrated not only the effectiveness but also the safety of the technology. ‘HoneySweet’ has been cleared for cultivation in the USA. By the appropriate regulatory agencies. The development and regulatory approval of ‘HoneySweet’ demonstrate the ability of RNAi technology to contribute to the sustainability of stone fruit production in regions impacted by PPV. Although it has taken almost 100 years since the identification of sharka, we are now able to effectively protect stone fruit species against this disease through the application of GE

    Phylogenetic position and virulence apparatus of the pear flower necrosis pathogen Ervvinia piriflorinigrans CFBP 5888(T) as assessed by comparative genomics

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    Erwinia piriflorinigrans is a necrotrophic pathogen of pear reported from Spain that destroys flowers but does not progress further into the host. We sequenced the complete genome of the type strain CFBP 5888T clarifying its phylogenetic position within the genus Erwinia, and indicating a position between its closest relative, the epiphyte Erwinia tasmaniensis and other plant pathogenic Erwinia spp. (i.e., the fire blight pathogen E. amyloyora and the Asian pear pathogen E. pynfoliae). Common features are the type III and type VI secretion systems, amylovoran biosynthesis and desferrioxamine production. The E. pinflonnigrans genome also provided the first evidence for production of the siderophore chrysobactin within the genus Erwinia sensu stricto, which up to now was mostly associated with phytopathogenic, soft-rot Dickeya and Pectobacterium species. Plasmid pEPIR37, reported in this strain, is closely related to small plasmids found in the fire blight pathogen E. amylovora and E. pynfoliae. The genome of E. pinflorinigrans also gives detailed insights in evolutionary genomics of pathoadapted Erwinia. (C) 2013 Elsevier GmbH. All rights reserved

    Effect of a pre-freezing treatment with cholesterol-loaded cyclodextrins on boar sperm longevity, capacitation dynamics, ability to adhere to porcine oviductal epithelial cells in vitro and DNA fragmentation dynamics

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    The aim of this work was to examine how a pre-freezing treatment with cholesterol-loaded cyclodextrins (CLC) affects boar sperm longevity, capacitation dynamics, ability to bind to a porcine telomerase-immortalised oviductal epithelial cell line (TERT-OPEC) in vitro and DNA integrity dynamics after freeze-thawing. Although the samples treated with CLC exhibited lower sperm quality than the control samples (P 0.05) after long-term incubation (26 h at 37 or 16 degrees C). Additionally, the CLC-treated spermatozoa underwent similar capacitation and DNA fragmentation dynamics as the control spermatozoa (P > 0.05). However, CLC-treated spermatozoa were better able to bind to TERT-OPEC in vitro (P < 0.0001). In conclusion, the pre-freezing treatment of boar spermatozoa with CLC enhanced the ability of the spermatozoa to bind to TERT-OPEC in vitro, which could have an effect on the establishment of the sperm reservoir in the ampullary-isthmic junction in vivo. Additionally, frozen-thawed spermatozoa can be stored at 16 degrees C for at least 6 h without a significant observable decline in sperm quality, which could be beneficial for the transport of thawed diluted doses of spermatozoa from the laboratory to the farm

    Development, Preimaginal Phases and Adult Sensillar Equipment in Aganaspis Parasitoids (Hymenoptera: Figitidae) of Fruit Flies

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    Aganaspis daci and Aganaspis pelleranoi (Hymenoptera: Figitidae) are important parasitoids of fruit flies. Here we studied, with light and scanning electron microscopy, aspects of their morphology that could help with plans to mass rear and thus contribute to improved pest control (preimaginal phases) and to shed light on parasitoid-pest relationships (sensillar equipment). The two species present a stalked egg, eucoiliform first and second-instar larvae and hymenopteriform third instar and mature larvae. The first instar presents tegumental differentiations in the mesoma and first metasomal segment in A. daci, but not in A. pelleranoi, while unlike other figitids, neither species displays setae in the mesosomal processes. Second and third instar and mature larvae present tegumental differentiations in A. daci, but not in A. pelleranoi. The moniliform (female) and filiform (male) antennae of A. daci and A. pelleranoi harbor seven types of sensilla, four of them (sensilla campaniformia, sensilla coeloconica type II, and two types of sensilla trichoidea) described here for the first time in Cynipoidea. The largest sensilla were the multiporous placoid sensilla, which were smaller and more numerous in A. pelleranoi. Species also differed to some extent in morphology of sensilla coeloconica. Observations on the ovipositor revealed the presence of coeloconic sensilla on Valva I in both species

    Acute toxicity in laboratory tests of fresh and aged residues of pesticides used in citrus on the parasitoid Aphytis melinus

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    California red scale Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae) is a worldwide citrus key pest. One of the control strategies currently promoted in the Mediterranean basin is the augmentative release of the parasitoid Aphytis melinus DeBach (Hymenoptera: Aphelinidae). A clear understanding of the impact that pesticides commonly used in citrus have on this parasitoid is therefore essential to assure the efficacy of the method. The aims of this study were to evaluate the lethal effects of freshly applied and laboratory-aged residues of eighteen selected pesticides recommended in IPM for citrus on A. melinus adults. According to their toxicity on A. melinus, the pesticides could be divided in four groups. Etofenprox and chlorpyrifos were harmful and persistent; abamectin was moderately harmful and slightly persistent; dimethoate, chlorpyrifos-methyl, and spirodiclofen were slightly harmful and moderately persistent; and azadirachtin, etoxazole, fenbutaestan, hexythiazox, mancozeb, petroleum oil, pirimicarb, propargite, pymetrozine, pyriproxyfen, spirotetramat, and tebufenpyrad were considered harmless. The information presented here will help us to integrate A. melinus release within an IPM for citrus in which pesticides are still used

    Response of Navel Lane Late citrus trees to regulated deficit irrigation: yield components and fruit composition

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    The effects of mid-summer regulated deficit irrigation (RDI) treatments were investigated on Navel Lane Late citrus trees over four seasons. Water restrictions applied from July until mid-September were compared with irrigation at full crop evapotranspiration (ETc). Two degrees of water restrictions were imposed: (1) RDI-1, irrigated at around 50% ETc and, (2) RDI-2, irrigated at 30-40% ETc. In addition, threshold values of midday stem water potential (Is) of -1.3 to -1.5 MPa for RDI-1 and of -1.5 to -1.7 MPa for RDI-2 were also taken into account. Results showed that Navel Lane Late is a citrus cultivar sensitive to water deficit since both RDI strategies reduced fruit size every year and water use efficiency in RDI trees was similar to control trees. However, the RDI-1 strategy allowed water savings up to 19% without reduction in yield when the water stress integral did not surpass 70 MPa day. RDI improved fruit quality, increasing total soluble solids and titratable acidity, while the fruit maturity was delayed. In conclusion, we suggest that RDI-1 strategy since it did not significantly impair the economic return can be applied in commercial orchards in case of water scarcity. Nevertheless, Navel Lane Late fruit is sensitive to water deficit and the fruit weight can be detrimentally affected

    Rabbit morula vitrification reduces early foetal growth and increases losses throughout gestation

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    Several studies have extensively examined structural and biochemical damage induced by cryopreservation that may lead to loss of rabbit embryo viability, but very little information is available on alterations in growth during gestation and at gene expression level. We started our work by comparing the distribution of losses of embryo and foetal development between control and vitrified rabbit morulae. Furthermore, data on foetal sack, foetal and maternal placenta and foetus size for 10-14 days of gestation were evaluated by ultrasonography. We reported that vitrification procedure causes detrimental effects on rabbit embryo and foetal development, with two major peaks of losses: one before the implantation (at day 6) and the other during the second part of gestation (after day 14). However, foetal loss may occur during the implantation process and placenta development, as there was a reduction in development of foetus produced from vitrified-warmed embryos between day 10 and 14 of gestation. For these reasons, using a recent microarray study performed in frozen-thawed rabbit embryos as a point of reference, we analysed the effects of vitrification procedure on the expression of 10 candidate genes in 6-day-old blastocysts obtained after vitrification and transfer. We observed that the relative expressions of mRNA transcripts from SCGB1A1, EMP1, ANXA3 and EGFLAM genes were significantly altered. This could help explain why a large number (29%) of vitrified embryos were successfully implanted but subsequently failed to develop to term. Further studies in subsequent embryo-foetal developmental stages, such as initiation of placenta formation, together with more sensitive high-throughput tools, should help us understand the deficiencies that hinder foetal development and identify the repairing mechanism employed by embryos to overcome vitrification effects. (C) 2013 Elsevier Inc. All rights reserved

    Natural enemies of the South American moth, Tuta absoluta, in Europe, North Africa and Middle East, and their potential use in pest control strategies

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    The South American tomato leafminer, Tuta absoluta Meyrick (Lepidoptera: Gelechiidae), is an invasive Neotropical pest. After its first detection in Europe, it rapidly invaded more than 30 Western Palaearctic countries becoming a serious agricultural threat to tomato production in both protected and open-field crops. Among the pest control tactics against exotic pests, biological control using indigenous natural enemies is one of the most promising. Here, available data on the Afro-Eurasian natural enemies of T. absoluta are compiled. Then, their potential for inclusion in sustainable pest control packages is discussed providing relevant examples. Collections were conducted in 12 countries, both in open-field and protected susceptible crops, as well as in wild flora and/or using infested sentinel plants. More than 70 arthropod species, 20 % predators and 80 % parasitoids, were recorded attacking the new pest so far. Among the recovered indigenous natural enemies, only few parasitoid species, namely, some eulophid and braconid wasps, and especially mirid predators, have promising potential to be included in effective and environmentally friendly management strategies for the pest in the newly invaded areas. Finally, a brief outlook of the future research and applications of indigenous T. absoluta biological control agents are provided

    Evaluation of the Susceptibility of Different Prunus Rootstocks to Natural Infection of Plum Pox Virus-T

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    Plum pox virus (PPV) has been observed in Turkey since 1968, but was not widespread except in apricot and plum trees in home gardens and ornamental parks in restricted areas. Susceptibility of six different Prunus rootstocks to strain PPV-T was assessed under natural inoculum pressure in the Izmir-Aegean region during 2010-2011. Aphid populations were monitored from the first week of April to the middle of June by the sticky-plant method one year after the rootstock plantation was established. Aphids collected from different rootstocks were tested individually by squash real-time RT-PCR and all rootstocks were regularly tested by DASI-ELISA. The largest aphid populations were observed at the end of May and the most abundant aphid species as averages over the two years were Myzus persicae (20.15%), Hyalopterus pruni (18.64%), Aphis craccivora (9.04%) and Aphis gossypii (8.36%). In 2011, the highest percentage of viruliferous aphids was found in M. persicae (34.78%), followed by H. pruni (32.50%), Macrosiphum euphorbiae (25.00%), A. gossypii (23.80%), A. spiraecola (12.50%) and A. craccivora (10.00%). Of the six Prunus rootstocks tested, only Nemaguard and Myrobalan 29C were infected by PPV-T, infection rate in 2010 being 6.0% (Nemaguard) and 4.0% (Myrobalan 29C). The infection rate increased to 16.0% for Nemaguard and 14.0% for Myrobalan 29C in 2011. However, the other rootstocks, Prunus marianna GF8.1, Docera6, GF677 and Garnem tested negative for PPV-T throughout 2011. PPV isolates obtained from naturally infected apricot trees (inoculum source) and from infected rootstocks in the experimental plot were characterized as PPV-T and had more than 99.5% nucleotide sequence identity

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