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Detection and classification of phytoplasmas mainly rely on amplification of the 16S rRNA gene followed by RFLP analysis and/or sequencing, because these organisms lack complete phenotypic characterization. Other conserved genomic loci have been exploited as additional molecular markers for phytoplasma differentiation. Two loci, SSU12p and LSU36p, selected by whole-genome comparison of 12 phytoplasma strains, were used for primer design, and were successfully tested on DNA samples from plants infected by phytoplasmas belonging to ten 16S ribosomal groups. The phylogenetic trees inferred from SSU12p and LSU36p loci were highly congruent to the trees derived from 16S rRNA and tuf genes of the same phytoplasma strains. Virtual RFLP analysis of the amplified SSU12p gene showed distinct patterns for most of the phytoplasma ribosomal subgroups tested. These results show that SSU12p and LSU36p genes are reliable additional markers for phytoplasma detection and differentiation.

Grapevine yellows (GYs) are phytoplasma-associated diseases worldwide distributed in grapevine growing areas. “Stolbur” phytoplasmas were reported in several areas and in almost all Italian regions, where their presence is constantly increasing. During the last years, the presence, identification and molecular characterization of phytoplasmas in symptomatic grapevine, was carried out in samples collected from diverse provinces of Apulia region (South Italy) in the frame of the mandatory program of surveillance for the quarantine phytoplasma disease “flavescence dorée”

Two of the most common leafhoppers present in Chile are the Cicadellidae Paratanus exitiosus and Bergallia valdiviana. They commonly occur in vineyards of central Chile, including some vineyards infected by phytoplasmas. The present study demonstrates that P. exitiosus and B. valdiviana can transmit 16SrIII-J phytoplasmas to grapevine and periwinkle plants. This provides improved understanding of the 16SrIII-J phytoplasma epidemiology in Chilean vineyards. EEA Mendoza Fil: Quiroga, Nicolás. Universidad de Chile. Facultad de Ciencias Agronómicas. Departamento de Sanidad Vegetal; Chile Fil: Longone, Maria Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza. Laboratorio de Fitopatología; Argentina Fil: González, Ximena. Universidad de Chile. Facultad de Ciencias Agronómicas. Departamento de Sanidad Vegetal; Chile Fil: Zamorano, Alan. Universidad de Chile. Facultad de Ciencias Agronómicas. Departamento de Sanidad Vegetal; Chile Fil: Pino, Ana María. Universidad de Chile. Facultad de Ciencias Agronómicas. Departamento de Sanidad Vegetal; Chile Fil: Picciau, Luca. Università di Torino. Dipartimento di Scienze Agrarie, Forestali e Alimentari (DISAFA); Italia Fil: Alma, Alberto. Università di Torino. Dipartimento di Scienze Agrarie, Forestali e Alimentari (DISAFA); Italia Fil: Paltrinieri, Samanta. Università di Bologna. Alma Mater Studiorum. Department of Agricultural and Food Sciences (DISTAL), Plant Pathology and Entomology; Italia Fil: Contaldo, Nicoletta. Università di Bologna. Alma Mater Studiorum. Department of Agricultural and Food Sciences (DISTAL), Plant Pathology and Entomology; Italia Fil: Bertaccini, Assunta. Università di Bologna. Alma Mater Studiorum. Department of Agricultural and Food Sciences (DISTAL), Plant Pathology and Entomology; Italia Fil: Fiore, Nicola. Universidad de Chile. Facultad de Ciencias Agronómicas. Departamento de Sanidad Vegetal; Chile

Three 16SrIII-J phytoplasma-positive vineyards were visited each 15 days from September 2017 to August 2018 to capture insects using entomological sweep nets and yellow sticky traps. Additionally weeds showing typical symptoms associated with the presence of phytoplasmas were also collected. The phytoplasma 16SrIII-J was detected for the first time in the leafhoppers Amplicephalus ornatus, A. pallidus and Bergallia sp., and in the weed Brassica campestris. Another phytoplasma classified as 16SrXI, was detected in one leafhopper belonging to the genus Paratanus. The identification of the 16SrXI ribosomal subgroup and the species of Bergallia sp. and Paratanus sp. are in progress, as well as the transmission trials with these two insects plus A. pallidus and A. ornatus.

In this work we developed a Smart Breath Analyzer device devoted to the tele-monitoring of exhaled air in patients suffering Chronic Obstructive Pulmonary disease (COPD) and home-assisted by mechanical ventilation. The device based on sensors allows remote monitoring of a patient during a ventilotherapy session, and transmit the monitored signals to health service unit by TCP/IP communication through a cloud remote platform. The aim is to check continuously the effectiveness of therapy and/or any state of exacerbation of the disease requiring healthcare. By preliminary experimental tests, the prototype was validated on a volunteer subject.

In this work we proposed a novel tele-medicine system called Smart Breath Network (SBN) with the perspective to improve the life of patients affected by a well-known respiratory disease called Chronic Obstructive Pulmonary Disease (COPD). The main therapy for COPD is home-assisted ventilation that requires fre- quent home intervention by specialized hospital staff in order to adjust dispensing and to verify the effectiveness of the therapy. Moreover, healthcare services need to improve the care of patients with difficult access to services, particularly those in rural or remote areas without sustaining high costs of service maintenance. The SBN system tries to fill the gap between healthcare services, that need to take care of a growing number of home-assisted COPD patients due to the today's alarming air pollution. The SBN network gathers all patient's respiratory data from Smart Breath Analyzers (SBAs) devices that are low-cost, simple to use, compact-sized devices that fit to almost any ventilator with bi-tube capabilities.

Biodiversity and ecosystem research deals with heterogeneous and distributed data resources generated from a large number of disciplines, which need to be integrated to advance the actual scientific knowledge. There is a clear need for integrative research, but at the same time, it is very challenging to provide appropriate solutions for the management of information. Within the biological and ecological informatics communities, recent efforts are devoted to the adoption of semantic web techniques, which provide a promising way to properly describe and interrelate different data sources to reduce barriers to data discovery and integration. In this respect, semantic harmonization, achieved with the development and use of thesauri, represents an essential precondition. The E-Biodiversity Research Institute of LifeWatch Italy (LW-ITA) developed thesauri, available as a web service (http://www.servicecentrelifewatch.eu/catalogue-of-services), on morphological traits of several groups of aquatic organisms, on alien and endemic species, and on genomics and barcoding. They were all implemented in Simple Knowledge Organization System (SKOS) and Resource Description Framework (RDF), using TemaTres, an open-source web-based platform for the collaborative management of thesauri, as well as a Linked Data interface and an RDF query language (SPARQL) endpoint. The development of LW-ITA thesauri is the result of an interdisciplinary collaboration of experts from both the specific domains and information and communication technologies. In this context, experts had a central role in the different phases of implementation of thesauri, resulting in the production of shared and stable versions of thesauri, and in the extension of the range of covered concepts.

Environmental Science is undergoing a period of big changes facing the challenge of data collection in an era driven by data. A big amount of data is pivotal when upscaling the study of ecological processes to the macro-ecological level, investigating multiple species distributed in multiple habitats. LifeWatch (LW), the European e-Science infrastructure on biodiversity and ecosystems, aims to strength the scientific research on biodiversity through the sharing biodiversity data making them Findable, Accessible, Interoperable and Reusable. According to this scenario, the LW Mediterranean Thematic Centre (MTC), together with the other TCs and the ICT members, developed a first case study collecting biodiversity data to investigate the vulnerability of ecosystems to biological invasions. The main scope of the Alien Species Show Case was assessing fragility of EUNIS habitat types to alien and invasive species and modelling their fragility to abiotic forcing. Occurrence records were gathered from specific datasets belonging to more than 30 public and private research institution. Overall a dataset on the occurrence of more than 12000 species from 560 sites (lagoon, marine, freshwater and terrestrial) across the Italian peninsula, was gathered. Records were organized and enriched with additional information to be useful to the further analysis. The data validation has been made through the data cleaning semi-automatic service in the LW dataportal, based on the lifewatch-ita Global Name Architecture, which refers to PESI, Worms, and COL. Here we report the steps that led to the show case development and the related results obtained, in terms of published scientific papers.

LifeWatch aims to develop Virtual Research Environments (VRE) for the study of biodiversity patterns and processes. The VRE should provide access to ICT services, data, software components but also provides a collaborative working environment where researchers can cooperate during all the research lifecycle. In the last years many VRE prototypes have been developed by different initiatives, providing an overview on advantages and disadvantages of different architectures. LifeWatch-Italy, within the framework of the alien species show case, explored the possibility to develop a workflow-oriented VRE, where researchers could harvest data from the LifeWatch dataportal and analyze them using modules embedding R script or calling external web services. A prototype of the workflow-oriented VRE (AS-VRE) has been made accessible through the LifeWatch service centre portal. In the AS-VRE researchers can replicate the statistical analyses employed in the alien species show case by using the Taverna software, a tool developed to orchestrate scientific workflows, that allows to connect tools, scripts and web services. A workflow-oriented VRE clearly can provide high flexibility, especially in term of the potential use of web services. However, this comes at the cost of a lower usability due to the complexity of orchestrating software that requires a minimum level of informatics skills, often lacking to researchers. Following a strategy proposed to overcome some usability issues, specific components have been developed to ease user interaction with the workflow execution. The AS-VRE currently allows researchers to produce a small bunch of analyses and represents a valuable starting point to think over VRE architectures and to build up a virtual research environment that will fit with important pillars such as usability, analytical power, easy access to data and software and cooperation improvement.

Biodiversity and ecosystem research is becoming central to major societal challenges as bioeconomy and climate action. To handle the scale and complexity of the scientific question being addressed, there is a strong need to integrate scientific knowledge. However, relevant data are heterogeneous, distributed and difficult to share. Initiatives to integrate data into viable resources for innovation in science and decision-making are being developed as local, regional, and global initiatives.The Data Portal of the LifeWatch Italy (LW-ITA) e-infrastructure aims at facilitating data sharing and stimulating collaborative and data-intensive science in the fields of biodiversity and ecosystem research. LW-ITA Data Portal functionalities ease data management through the Data Life Cycle (quality assurance and quality control, metadata creation, integration, data discovery, visualization and analysis). We present the currently available solutions for data management and sketch future developments. We aim to create a service-oriented e-infrastructure for biodiversity and ecosystem data, where all components are accessible through web services, in order to provide one single entry point to existing biodiversity and ecosystem data resources in Italy and to make them findable, accessible, interoperable and re-usable (FAIR principles) at all levels, from local to global.

Kiwifruit bleeding sap samples, collected in Italian and Chilean orchards from symptomatic and asymptomatic plants, were evaluated for the presence of Pseudomonas syringae pv. actinidiae, the causal agent of bacterial canker. The saps were sampled during

The diversity of European stone fruit yellows (ESFY) phytoplasmas was studied using a multilocus typing approach to verify possible variability related to phytoplasma population modification/diversification. After their identification at the 16S ribosomal level about 70 ESFY strains were selected for further characterization using amplicons from ace, imp and secY genes as molecular markers. Field collected samples from diverse stone fruits, reference strains in periwinkle before micropropagation or micropropagated since up to 15 years, and a strain from pineapple were employed. Characterization was performed by RFLP analyses and sequencing of selected amplicons obtained in nested PCR assays. A clear differentiation of the strain infecting pineapple in Colombia from all others was observed by both, differential restriction profiles on all the three genes and SNPs presence in the ace gene sequence alignment. The RFLP analyses of strains from field collected material showed that the profiles of strains before micropropagation correspond to four out of the 18 different lineages detected in the field-collected material. Moreover also the strains maintained in micropropagation were differentiable from the corresponding strains before micropropagation, and resulted differently represented in the field-collected samples. In particular only one lineage was consistently retrieved in all the different materials, while one lineage was present only in micropropagated strains and another in micropropagated and field collected samples. This is a clear indication of rearrangement in those genes in the diverse samples according to differential plant host (pineapple, stone fruits, periwinkle), growing system and time frame. In particular the pineapple strain showed differential RFLP profiles on ace and secY genes, in the first case confirmed also by the presence of SNPs in the sequence, indicating possible adaptation of this strain to the new host plant in which ESFY phytoplasma was detected for the first time.

En este artículo los autores señalan la necesidad de desarrollar un modelo de prevención de PSA que reduzca la diseminación del cancro bacteriano. Así mismo explica un modelo de detección desarrollado en Italia y experimentado en Chile que se basa en el muestreo y análisis del “lloro primaveral” del kiwi, es decir, del flujo xilemático que se libera por los cortes de poda de las plantas.

Lagoons are ecotone ecosystems, naturally exposed to immigration from their freshwater and marine input environments; strictly lagoonal species are rare, if ever occurring, and lagoon community are likely to be organised through loItery competition processes. These evidences suggest the study of lagoons as models address ecosystem fragility to alien species. Here, we present an analysis of lagoon guild fragility to alien species carried out using the e-Science facilities of LifeWatch, the European Research Infrastructure on biodiversity and ecosystem research. The analysis has been performed on two EUNIS habitat (X02 and X03) using data existing on 18 Italian lagoons. The analysis has been carried out taking into account that rarity, redundancy and singularity are key properties of different guilds in lagoon ecosystems at every geographical area, affecting B (Beta) and y (Gamma) diversity. At every area an high regional biodiversity is determined by a large number of rare species and a high dissimilarity among lagoons. Life cycle traits and the behaviour of larval stages, at the species level, as well as lagoon openness and vigour, at the ecosystem level, seem to have a major role to explain the difference in patterns of biodiversity between study areas at a biogeographical scale. The same species and ecosystem level properties, together with spatial patchiness, seem also to be key factors downscaling biodiversity analysis at the landscape level.

Phytoplasmas found in Chilean grapevines showing yellows symptoms were identified as belonging to the ribosomal subgroups 16SrI-B and 16SrI-C (‘Candidatus Phytoplasma asteris’), 16SrIII-J (X-disease group), 16SrV-A (‘Ca. P. ulmi’), 16SrVII-A (‘Ca. P. fraxini’), 16SrXII-A (stolbur or “bois noir”) (Gajardo et al., 2009; González et al., 2010). The presence of these pathogens in the plants depends on both propagation of infected plants and spreading by different insect species which feed on grapevine and also on the weeds growing near and/or in vineyards. In infected vineyards, several insects belonging to the family Cicadellidae positives to phytoplasmas were found. The most common was Paratanus exitiosus (Beamer) in which phytoplasmas of 16SrI-B, 16SrIII-J, 16SrVII-A, and 16SrXII-A subgroups were detected. In the present work, we verified the phytoplasma transmission ability of the leafhopper P. exitiosus. Materials and Methods During 2011 (since September till December) and 2012 (since January till May) leafhopper survey was carried out in Chilean vineyards infected by phytoplasmas, two located in Metropolitana Region (1 and 2) and one in Valparaiso Region (3), to know biological characteristics of P. exitiosus and its ability to transmit phytoplasmas. The insects were captured by sweeping with an entomological net. During the sampling period adults of P. exitiosus captured have been released into entomological cages to let them feed on three plants of periwinkle (Catharanthus roseus (L.) G. Don) grown from seed and previously tested to ascertain the absence of phytoplasmas. A total of 81 plants were used. Periwinkle plants were tested starting five months after transmission trials, while dead insects were then preserved in 70% ethanol. Insects and periwinkles were tested in order to identify the phytoplasma presence. Total nucleic acids (TNAs) were extracted with chloroform/phenol methods, dissolved in Tris-EDTA pH 8.0 buffer, and maintained at 4°C; 20 ng/µl of nucleic acid were used for amplification. After direct PCR with primer pair P1/P7, nested PCR with R16F2n/R2 primers (Gundersen and Lee, 1996) was performed. PCR and nested PCR reactions were carried out following published protocol (Schaff et al., 1992). P1/P7 amplicons were purified using Concert Rapid PCR Purification System and DNA fragments were cloned. Putative recombinant clones were analyzed by colony PCR. Selected fragments from cloned DNAs were sequenced in both directions using the BIG DYE sequencing terminator kit. The sequences were then aligned with BLAST engine for local alignment (version Blast N 2.2.12). Identification was done using in silico restriction fragment length polymorphism (RFLP) analyses on sequences amplified with primer pair R16F2n/R2 with BstU1 and HhaI restriction enzymes (Wei et al., 2007). Results and Discussion In the vineyard 1 the P. exitiosus capture rate remained constant during all months, with a decrease in October 2011 and April 2012. In the vineyard 2 the highest number of individuals was obtained in December 2011 and January 2012. In the vineyard 3 P. exitiosus was less abundant in September 2011, remained constant during the other months and increased only during February 2012. These results indicate that P. exitiosus tends to be more abundant during summer time. Three out of 81 periwinkles used for transmission trials were positive to phytoplasmas. Two (VC28C and VC31C) correspond to the transmission trials carried out with insects captured in the vineyard 1 in two different months (November and December 2011 respectively), the third (VC33A) was infected from insects captured in the vineyard 3 during December 2011. Cloned P1/P7 fragments were sequenced and there was no sequence difference between the cloned fragments from the three periwinkles (1,819 bp). The similarity percentages of VC28C, VC31C, and VC33A phytoplasmas, showed a close correlation (99.6, 99.7, 99.6 % respectively) with the strain Ch10 (AF147706...