First Report of CRISPR/Cas9 Gene Editing in Castanea sativa Mill

CRISPR/Cas9 has emerged as the most important tool for genome engineering due to its simplicity, design flexibility, and high efficiency. This technology makes it possible to induce point mutations in one or some target sequences simultaneously, as well as to introduce new genetic variants by homology-directed recombination. However, this approach remains largely unexplored in forest species. In this study, we reported the first example of CRISPR/Cas9-mediated gene editing in Castanea genus. As a proof of concept, we targeted the gene encoding phytoene desaturase (pds), whose mutation disrupts chlorophyll biosynthesis allowing for the visual assessment of knockout efficiency. Globular and early torpedo-stage somatic embryos of Castanea sativa (European chestnut) were cocultured for 5 days with a CRISPR/Cas9 construct targeting two conserved gene regions of pds and subsequently cultured on a selection medium with kanamycin. After 8 weeks of subculture on selection medium, four kanamycin-resistant embryogenetic lines were isolated. Genotyping of these lines through target Sanger sequencing of amplicons revealed successful gene editing. Cotyledonary somatic embryos were maturated on maltose 3% and cold-stored at 4°C for 2 months. Subsequently, embryos were subjected to the germination process to produce albino plants. This study opens the way to the use of the CRISPR/Cas9 system in European chestnut for biotechnological application

Trimester-specific reference intervals for thyroid function parameters in pregnant Caucasian women using Roche platforms: a prospective study

Background: Standard thyroid function parameters reference intervals (RI) are unsuitable during pregnancy, potentially resulting in incongruous treatments that may cause adverse effects on pregnancy outcomes. We aimed at defining trimester-specific TSH, FT4 and FT3 RI, using samples longitudinally collected from healthy Caucasian women. Materials and methods: Blood samples from 150 healthy Caucasian women, who had a physiological gestation and a healthy newborn at term, were collected in each trimester and at around six months post-partum. They showed mild iodine deficiency. After excluding women with overt TSH abnormalities (> 10 mU/L) and/or TPO antibodies, data from 139 pregnant women were analyzed by means of widely used Roche platforms, and TSH, FT4 and FT3 trimester-specific RI were calculated. Post-partum data were available for 55 subjects. Results: Serum TSH RI were 0.34-3.81 mU/L in the first trimester, and changed slightly to 0.68-4.07 U/L and 0.63-4.00 mU/L in the second and third trimester, respectively. Conversely, both FT4 and FT3 concentrations progressively decreased during pregnancy, the median values in the third trimester being 14.8% and 13.2% lower, respectively, than in the first trimester. Thyroid function parameters in the first trimester were similar to those measured after the end of pregnancy. Conclusions: This study calculates trimester-specific RI for thyroid function parameters in pregnancy, and proposes the reference limits that should be adopted when using Roche platforms in Caucasian women

Il sistema di monitoraggio sismico dell’Osservatorio Vesuviano – INGV

Obiettivo principale del monitoraggio dei vulcani attivi è individuare e misurare fenomeni che possono essere indotti dal movimento del magma in profondità. Dal punto di vista sismologico questi fenomeni possono essere sciami sismici, eventi a bassa frequenza, microtremore vulcanico ed eventi very long period (VLP). Attraverso la misura, l'analisi e la corretta interpretazione di questi fenomeni è possibile capire in anticipo se un vulcano sta evolvendo verso una ripresa dell'attività eruttiva. L'Osservatorio Vesuviano - INGV ha tra i suoi compiti istituzionali il monitoraggio del Vesuvio, dei Campi Flegrei e di Ischia che sono, come è noto, vulcani a alto rischio a causa del loro stile eruttivo prevalentemente esplosivo e della presenza nelle loro prossimità di vaste aree urbanizzate. Per effettuare il monitoraggio sismologico di dette aree l'Osservatorio Vesuviano ha sviluppato e mantiene una rete che trasmette i dati in continuo al centro di sorveglianza. La configurazione attuale della rete comprende 28 stazioni analogiche a corto periodo (1Hz) e 4 stazioni digitali a larga banda.PublishedRoma1.4. TTC - Sorveglianza sismologica delle aree vulcaniche attiveope

Oleosin Cor a 15 is a novel allergen for Italian hazelnut allergic children

BACKGROUND: Hazelnut allergy, which is characterized by symptoms that range from mild to severe, is one of the most common allergies in children throughout Europe, and an accurate diagnosis of this allergy is therefore essential. However, lipophilic allergens, such as oleosins, are generally underrepresented in diagnostic tests. We therefore sought to characterize the IgE reactivity of raw and roasted hazelnut oleosins, using the sera of hazelnut‐allergic pediatric patients. METHODS: Raw and roasted hazelnut oil body–associated proteins were analyzed by means of 1D and 2D electrophoresis and MS. Oleosin IgE reactivity was assessed by immunoblotting with the sera of 27 children who have confirmed hazelnut allergies and from 10 tolerant subjects. A molecular characterization of the oleosins was performed by interrogating the C. avellana cv. Jefferson and cv. TGL genomes, and through expression and purification of the recombinant new allergen. RESULTS: A proteomic and genomic investigation allowed two new oleosins to be identified, in addition to Cor a 12 and Cor a 13, in hazelnut oil bodies. One of the new oleosins was registered as a new allergen, according to the WHO/IUIS Allergen Nomenclature Subcommittee criteria, and termed Cor a 15. Cor a 15 was the most frequently immunorecognized oleosin in our cohort. Oleosins resulted to be the only immunorecognized allergens in a subgroup of allergic patients who showed low ImmunoCAP assay IgE values and positive OFC and PbP. Hazelnut roasting resulted in an increase in oleosin immunoreactivity. CONCLUSION: A novel hazelnut oleosin, named Cor a 15, has been discovered. Cor a 15 could play a role in eliciting an allergic reaction in a subgroup of pediatric patients that exclusively immunorecognize oleosins. The high prevalence of hazelnut oleosin sensitization here reported further confirms the need to include oleosins in routine diagnostic procedures

Spring 1984 Conference Issue

Research Update (page 3) Low Management Kentucky Bluegrass Cultivar Trials (5) Nitrogen Leaching Losses from N Fertilizer Applied to Turf (8) Fifty-third Annual Turf Conference and Eighth Industrial Show (10) Liquid Versus Dry Nitrogren Fertilizer Applications on Kentucky Bluegrass (13) Growth Retardant, Embark (16) Fungicides: The Good, The Bad and The Ugly (18

Structural, physiognomic and aboveground biomass variation in savanna-forest transition zones on three continents. How different are co-occurring savanna and forest formations?

Through interpretations of remote sensing data and/or theoretical propositions, the idea that forest and savanna represent "alternative stable states" is gaining increasing acceptance. Filling an observational gap, we present detailed stratified floristic and structural analyses for forest and savanna stands mostly located within zones of transition (where both vegetation types occur in close proximity) in Africa, South America and Australia. Woody plant leaf area index variation was related in a similar way to tree canopy cover for both savanna and forest with substantial overlap between the two vegetation types. As total woody plant canopy cover increased, so did the contribution of middle and lower strata of woody vegetation to this total. Herbaceous layer cover also declined as woody cover increased. This pattern of understorey grasses and herbs being progressively replaced by shrubs as canopy closure occurs was found for both savanna and forests and on all continents. Thus, once subordinate woody canopy layers are taken into account, a less marked transition in woody plant cover across the savanna-forest species discontinuum is observed compared to that implied when trees of a basal diameter > 0.1m are considered in isolation. This is especially the case for shrub-dominated savannas and in taller savannas approaching canopy closure. An increased contribution of forest species to the total subordinate cover is also observed as savanna stand canopy closure occurs. Despite similarities in canopy cover characteristics, woody vegetation in Africa and Australia attained greater heights and stored a greater concentration of above ground biomass than in South America. Up to three times as much aboveground biomass is stored in forests compared to savannas under equivalent climatic conditions. Savanna/forest transition zones were also found to typically occur at higher precipitation regimes for South America than for Africa. Nevertheless, coexistence was found to be confined to a well-defined edaphic/climate envelope consistent across all three continents with both soil and climate playing a role as the key determinants of the relative location of forest and savanna. Taken together these observations do not lend support the notion of alternate stable states mediated through fire-feedbacks as the prime force shaping the distribution of the two dominant vegetation types of the tropical lands