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After the recent high-impact European outbreaks of Xylella fastidiosa (Xf), a xylem-limited plant pathogenic bacterium native to the Americas, this research aims to rank the risks of potential entry, establishment and spread of Xf in new countries across Europe, the Middle East and North Africa. A novel risk-ranking technique is developed, based on combining entry risk drivers (imported plants, direct flights and ferry connections) with risk factors related to establishment and spread (presence of potential insect vectors, vulnerable economic crops, alternative hosts and climate suitability) of this pathogen. This reveals that western European countries have the highest risk for entry, but that the Mediterranean basin runs the highest risk for establishment and spread of Xf. Lebanon in particular has the highest level of risk for Xf dispersal within its suitable territory. Countries without current outbreaks combining high risks of Xf arrival and establishment are mainly in the Mediterranean basin: Turkey is at the highest level of risk, followed by Greece, Morocco and Tunisia, which are ranked at the high level. The ranking model also confirms the vulnerability, in terms of invasion by Xf, of southern European countries (Italy, Portugal and Spain) in which the pathogen has already been reported. High summer temperatures in these southern countries are likely to be the significant determinant for the overall invasion process, while northern European countries have a high level risk for the arrival of the pathogen, but relatively low summer temperatures may limit establishment and spread of major outbreaks. In general, our study provides a useful approach for mapping and comparing risks of invasive non-native species and emerging pathogens between countries, which could be useful for regional horizon scanning and phytosanitary and biosecurity management.

As olive is the most widespread fruit tree in Montenegro, eventual detection of the bacterium Xylella fastidiosa would cause great economic losses, besides implying the implementation of a permanent monitoring campaign. This is especially due to the importance of early detection of the bacteria as well as its vector(s) for the timely implementation of control measures to prevent it spreading. Given the significance of the vectors in the transmission of X. fastidiosa and the limited information about their presence in Montenegro, a study was undertaken to determine the species of Auchenorrhyncha (Cicadomorpha) present in olive groves on the Montenegrin coast. During 2018, insects were collected using sweep net sampling at three locations in the area of Valdanos and two locations in the area of Radanovići (Sampling was carried out on weeds, twice in the area of Valdanos (late May and late October) and once in the area of Radanovići (early September). The collected insects were preserved in 96% alcohol until determination. Insect identification was performed on the basis of the morphological characteristics of the species. Sixteen Auchenorrhyncha species were identified of which 12 belong to the Cicadomorpha, three to the Aprophoridae and nine to the Cicadellidae families. In all three locations in the area of Valdanos the most prevalent was Philaenus spumarius L. (main vector of X. fastidiosa in Europe) while Lepyronia coleoptrata (L.) was mainly collected in the area of Radanovići

Xylella fastidiosa is able to colonise a very large number of plant species, but considering each subspecies/phylogenetic clade the number of associated susceptible hosts is significantly reduced. Although strains genetically related most likely share similar host range, using phylogenetic relationships to infer information regarding the potential hostrange of new strains is still problematic, and pathogenicity tests remain the only means to assess the capability of a given strain to infect or not a specific plant species. Based on the current European legislative provisions on X. fastidiosa, information on the host range of the strain(s) causing an outbreak have regulatory consequences. In this context, we have made efforts to prove experimentally the capability of X. fastidiosa subsp. pauca, ST53, one of the most virulent European genotypes, to infect grapes, using 23 grape varieties (Vitis vinifera) and four rootstocks. Upon needle inoculation, plants were monitored for 18 months, using standand diagnostic methods (qPCR and isolation), supported by vector-transmission tests and observation of thin sections of the inoculated stems, stained using the LIVE/DEAD BacLight kit. qPCR assays on samples collected at the inoculation points (i.p.) and from the distal portions, 6 and 12 months post-inoculation, yielded positive reactions in more than 90% of the i.p., whereas in half of the cultivars scattered amplifications (the majority yielding Cq values > 30) occurred in some replicates at 15–20 cm from the i.p., but none of the apical portions tested positive. Isolations made 18 months after the inoculation, either from mature leaf petioles and stems portions harbouring the inoculation points, failed to recover actively growing colonies. At the same time, microscope observation of the thin sections showed only the presence of aggregates of dead Xylella-cells at the i.p. Transmission tests performed using specimens of Philaenus spumarius caged on the inoculated grapes produced negative results for both insects and recipient plants. The overall results showed that the bacterium was successfully delivered into the stem of the grapes and bacterial residual could be qPCR-detected even one year after the inoculation, but noneof the inoculated cultivars sustained active bacterial multiplication and colonisation.

The difficulties in the implementation of containment measures over five years of a severe epidemic of Xylella fastidiosa in Salento (Apulia, southern Italy), as a consequence of uncontrolled free dissemination of fake news, demonstrated that timely and effective information and communication on the same ground utilised by the misinformation, social media, are strategic not only in contrasting the bacterium/vectors on the outbreaks but especially for prevention. From 2016 Infoxylella®, a spontaneous initiative of volunteers (the editorial staff include scientists, olive growers and agronomists), considering the complexity of scientific arguments, the continued rapid evolution of legislation and the confusion created by misinformation, try to inform public opinion and media with simple language, filtering the essential information only from official verified sources and profiting from a continuous collaboration with the coordination team of the H2020 projects POnTE and XF-ACTORS. Thirty-four municipalities, producer's organisations, associations, companies, etc. became formal supporters of Infoxylella, now in the process of becoming a non-profit organisation. Besides the growing popularity and success on the web and social media (i.e. the Facebook page called Infoxylella.it reached 10,000 followers in April 2019), Infoxylella became a reliable source of information for journalists (mentioned asa source in over 60 articles in national journals), other media and in particular is active in the territory participating in informative seminars organised by the municipalities (over 30 in 2018). In order to involve the local administrations in prevention and public information as collective contrast to the epidemic, Infoxylella prepared an informative booklet already adopted, for printing or publishing in electronic format, by 25 local entities. Despite its young age and the specificity on Xylella in Apulia, Infoxylella.it could be considered as a case study providing useful practical experience and ideas to improve elsewhere public awareness on phytosanitary issues and direct involvement of population in the contrast of other epidemics.

Early detection of infections in programs of production of virus-tested vine propagating material is of paramount importance to reduce costs of maintaining sanitized plants. In Apulia (Italy), the Regional Project “Re.Ge.Vi.P.”, aiming to the recovery, conservation and sanitary improvement of autochthonous vine cultivars, gave the cue to assess the minimum period of time required after sanitation to detect virus infections with molecular techniques. Previous results of this work, relatively to the period 2013-2015, were given (Morelli et al., 2015) and regarded the analysis of the sanitary status of ca. 150 grape cultivars from which ca. 100 apexes were obtained by thermotherapy and in vitro culture. The work, continued in 2016-2017, allowed to confirm the previous findings which showed the good efficacy of the sanitation protocols and the reliability of applied diagnostic techniques in the whole process of production of vine propagating material.

Currently Apulia region accounts about 59 winegrape cultivars authorised for growing, including historical varieties. The existence of autochthonous minor varieties in the region has been recognized and is recently at the centre of a program aiming to their recovery and their further exploitation for commercial use. In the present study, 87 traditional, autochthonous and minor Apulian grapevine accessions were examined for their genetic profiles. Among them, 64 unique genotypes were found. Synonyms among Apulian genotypes and Italian varieties cultivated in Apulia or other Italian regions were identified by searching for genetic matching in different molecular database publicly available and through bibliographic search with published genetic data. The main goal of the current study was to identify, characterise and catalogue local germplasm rich of economical and historical significance.

Montenegro, as candidate Country for the accession in the European Union, in the framework of the procedure for alignment and together with the harmonization of national legislation with CMO Wine, decided to conduct a new viticultural zoning study at national level. Within the overall objective of further strengthen the viticulture and wine sectors through the harmonization with EU standards, the IPA project "Technical Support to Renewal of viticulture zoning of Montenegro" (Europe Aid/136071/DH/SER/ME) had several other specific objectives including: a) the establish a new Viticulture Zoning as precondition to set up a system of designations for wine with Geographical Indications (PDO/PGI system); b) the official classification of the viticultural areas of Montenegro in climatic zone as required by EU regulation i.e. for the adoption of the oenological practices and defining the restrictions in their application. The two years project started in April 2015, leaded by CRSFA "Basile Caramia", was carried out by a multidisciplinary team of Italian and Serbian experts with the collaboration of experts from Montenegro. Starting from an old viticultural zoning of former Yugoslavia carried out in 1976, a huge amount of different existing data, provided by the Ministry of Agriculture and Rural Development and other Institutions and wineries of Montenegro, was collected, processed and analysed. For the climate analysis, data from 12 meteorological stations and for three periods, one 44-years long (1971-2014) and two 20-years long (1971-1990 and 1995-2014), were used to calculate 11 bioclimatic indexes. Scientific literature, soil maps of Montenegro, the official vineyard and winery registers, the chemical analysis of all wines on the market in the last 5 years, different type of digital maps and DEM were the main source of data used. Additional primary data, used to confirm the existing ones, was collected in 19 vineyards/locations, selected as representatives of the candidate homogeneous subregions, by opening soil profiles, sampling, analyzing and comparing soils, grapes (2015 and 1016) and wines (2015) of the main cultivars. From the analysis of the vineyard parcels the main characteristics and statistics for each homogeneous area, including their borders were described in detail and represented, using contemporary advanced GIS methods (GIS software / ESRI ArcGIS etc.), in thematic maps. The results of the study confirm that Montenegro has, in a relatively small territory, very different terroirs particularly due to a complex orography and the Mediterranean climate strongly influenced by Adriatic Sea and the big Skadar lake as well as, locally, high mountains and rivers. The zoning study identified three main homogeneous viticultural areas or regions and eleven sub-regions that, classified among II (B), IV (C2) and V (C3) zones, have been formally recognized published on National Gazette: Crnogorski basen Skadrskog jezera (sub-regions Bjelopavli?i, Crmnica, Katuni, Podgorica, Rijeka, Piperi), Crnogorsko Primorje (sub-regions Boka kotorska, Budva-Bar, Ulcinj, Jadransko zale?e) and Nudo. Additionally, based just on climate analysis and perspections, five small new potential viticultural areas where identified within the area Sjever: Bijelo Polje, Dragaljsko Polje, Nik?i? and ??epan Polje. Those places, in the past too cold for viticulture, due to the ongoing and observed climate change, seem to be suitable for grape growing and should be tested experimentally. The zoning represent at the same time an instrument of deep knowledge about viticulture and territory available for the Montenegrin producers and the technical framework for the establishment of a new quality wine system based on the certification of the geographical origin. The final technical elaborate of the study, consisting in around 1.200 pages full of data, information and 22 different thematic maps for each area, will help the producers and their organizations/consortia in preparing the technical dossier than accelerating the procedures for the official recognition and registration of new PDO/PGI wines.

INTRODUCTION Grapevine leafroll disease (GLRD) is one of the most diffused among viral diseases for the grapevine (Vitis vinifera L.) (Almeida et al., 2013; Maare et al., 2013). To date, 11 Grapevine leafroll associated viruses (GLRaVs) have been described: GLRaV-1, -2, -3 -4, -5, -6, -7, -9, -Pr, -De, and GLRaCV, all belonging to the Closteroviridae family. Of these viruses, GLRaV-2 is the only member of the Closterovirus genus; a tentative genus called Velavirus has been created to classify GLRaV-7, while the rest of GLRaVs belong to the Ampelovirus genus. GLRaV-1 and GLRaV-3 are apparently the most common ampeloviruses in vineyards and, together with GLRaV-2, the main viruses responsible for GLRD damages. The others ampeloviruses (GLRaV-4, -5, -6, -9, -Pr, -De, and GLRaCV) belong to a common phylogenetic clade (subgroup II) in the Ampelovirus genera and may be classified as divergent variants of one virus, GLRaV-4, instead of distinct species, according to recent taxonomical classification (Martelli et al., 2012). The presence of some GLRaVs were reported from many viticultural areas in Turkey in the past. GLRaV-4 (Kaya et al., 2012) and -5 (Buzkan et al., 2010), the first GLRaV-4-like viruses, have been lately detected in some locations in Turkey. Therefore, a study was carried out to investigate the presence of GLRaV-9 in the subgroup II in Turkish autochthonous varieties and potential vector insects. MATERIALS AND METHODS A total of 116 vineyards in two viticultural areas (eastern Mediterranean and southeast Anatolia) was visited for the presence of leafroll-like symptoms and mealybug vector investigation. Grapevines were essentially self-rooted, table varieties. Plant tissues and Planococcus ficus samples were collected from July to the end of August and were stored at +4oC for PCR (mealybugs preserved in 70% ethanol solution). About 100 mg of leaves/petioles was processed for total nucleic acid (TNA) isolation with silica-capture method (Foissac et al., 2005). Five or ten insects, depending upon the size, were also used for TNA isolation (Singh et al., 1995). All TNAs from plants and insects were stored at -20oC before cDNA synthesis. Two-step reverse transcription polymerase chain reaction (RT-PCR) was performed using primers LR9-F/LR9-R (Alkowni et al., 2004). PCR amplicons were custom-sequenced directly with both primers by Medsantek (Turkey). Alignments of the obtained sequences with additional homologous sequences retrieved from GenBank after using the Blastn program (Altschul et al., 1997). A Neighbor-Joining method (Saitou and Nei, 1987) with bootstrap validation was applied to draw a phylogenetic dendrogram (from MEGA6 package). RESULTS AND DISCUSSION A total of 423 grapevine samples from 33 autocthonous varieties was tested by RT-PCR assay. GLRaV-9 was detected in 16 samples from both regions. The highest number of GLRaV-9 infected sample was obtained from southeast Anatolia region (approx. 3%). Comparative analysis of 10 Turkish (TK) and other GLRaV-4-like isolates showed close relationship. Two clustering patterns could clearly be observed in the phylogenetic tree (Fig. 1). Group one contained five TK isolates (TK 51, 53, 54, 55, 65) from three autochthonous varieties (cv. Yalova incisi, Pafu and Hönüsü) from different locations and their nucleotide identity ranged between 82% to 85% with those of GLRaV-4 strain 9 (KJ810572, AY297819). Four TK isolates grouped with the US GLRaV-4 isolate (FJ467503) (Abou-Ghanem Sabanadzovic et al., 2012) in the same cluster (Group II). The nucleotide identity of the isolate TK48 and the US GLRaV-4 isolate was 97%. The TK 78 remained out of two clusters having nucleotide identity as one of other GLRaV-4 variants. To our knowledge, This is the first report of GLRaV-9 in turkish vineyards. PCR reaction with all the P. ficus samples did not result positive for the presence of GLRaV-9. Plant samples from the same mealybug infested areas were also negative for GLRaV-9 detection.

Early detection of infections in programs of production of virus-tested vine propagating material is of paramount importance to reduce costs of ma int aining san itized plants. In Apulia (ltaly), the Regional Project "Re. Ge .Vi. P." , aiming to the recovery, conservation and sanitary improvement of autochthonous vine cultivars, gave the cue to assess the minimum period of time required after sanitati on to detect virus infections with molecular techniques. Previous results of this work, relatively to the period 2013 -2015, were given (Morelli et al. , 2015) and regarded the analys is of the sanitary status of ca. 150 grape cultivars from which ca. 100 apexes were obtained by thermotherapy and in vitro culture. The work, continued in 2016- 20 17 , allowed to confirm the previous findings which showed the good efficacy of the sanitation protocols and the reliability of applied diagnostic techniques in the whole process of production of vine propagating material.

The ambitious idea for the development of new tablegrape seedless varieties, suitable for growing in the Mediterranean basin and highly competitive on the international markets, has recently led to the foundation (in 2015) of the Italian Variety Club (IVC) a network of 17 enterprises and 2 research Institutions in Southern Italy. This synergic collaboration among private companies and researchers aims to develop and transfer varietal and technical innovation to the growers and to strengthen the Italian table grape supply chain merging high quality products and sustainable agriculture. The breeding program is organized in different interconnected activities such as: "seed x seedless" varieties and "seedless x seedless" varieties controlled crosses, involving Apulian traditional genotypes and élite cultivars; F1seedling production with molecular marker assisted selection (MAS) for an early identification of seedless genotypes; in vitro embryo culture; plant evaluation under field conditions. In the coming years the activities of varietal description and official registration, Patent protection, and establishment of stock fields will be initiated. As result of the 3-year activities, the progenies of 75 "seed x seedless" varieties crosses, 28 "seedless x seedless" varieties crosses and 12 crosses involving germplasm with resistance traits have been obtained and analysed and the field evaluation begun. With regard to progenies derived from "seedless x seedless" varieties crosses, the average embryogenesis percentages (no. of germinated embryos/no. of cultivated ovules) was about 8-9%, varying according to the parents used. Following the extensive use of MAS about 40% of each "seed x seedless" varieties progeny carried allele associated to seedless locus. Actually about 6,000 new genotypes are grown in the first selection field, while further 6,000 seedless plants are going to be planted within 2018.

In Italy, grapevine certification system requires the status of “virus controlled” for multiplication and commercialization. This system entails free plant propagation material from Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 1, 2, 3 (GLRaV-1, -2, -3), Grapevine fanleaf virus (GFLV), Grapevine fleck virus (GFkV) and Arabis mosaic virus (ArMV)]. Routinely applied bio- and molecular assays for the detection of systemic plant pathogens, such as wood indexing, serological and (RT-) PCR molecular tests, are time-consuming and always require a priori knowledge of the target. On the other hand, the availability of High-Throughput Sequencing (HTS) techniques, which are free from the abovementioned constraints, allows the characterization of the “virome” of the analysed plants. Indeed, data originating from HTS give an unbiased picture of the virus/viroid content of a plant. In this frame, a task of the Apulian Regional Project Re. Ge. Vi. P (Recovery of Apulian grape germplasm) aims to assess the sanitary status of 20 certified autochthonous grapevine clones, table and wine cultivars, by HTS. In 2016, small RNAs (sRNAs) libraries were synthesized from 20 grape accessions and subjected to Illumina sequencing. The downstream data were processed using a custom-developed bioinformatics pipeline including the quality control of the libraries, the de novo assembly of the reads using Velvet assembler and the Blast annotation of the obtained contigs. Annotated contigs confirmed the absence of the above-regulated viruses. Also, this approach showed the presence of two viruses and two viroids not targeted in routinely performed assays, i. e. Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine rupestris vein feathering virus (GRVFV), Hop stunt viroid (HSVd) and Grapevine yellow speckle viroid 1 (GYSVd-1). These results, confirmed by RT-PCR assays, strengthen the potentiality of HTS approach in plant “virome” definition and its application in grapevine certification schemes.

Osservazioni e rilievi di campo integrati da un programma di campionamenti e saggi diagnostici di laboratorio confermano in alcune delle varietà oggetto di studio una minore incidenza delle infezioni di Xylella fastidiosa, a cui sono associate sintomatologie di deperimento e manifestazioni di disseccamento più lievi e attenuate rispetto a quanto osservato in cultivar definite altamente suscettibili, ove la presenza di infezioni di X. fatidiosa compromette la sopravvivenza stessa delle piante. I risultati, oltre a confermare i fenomeni di resistenza già osservati in precedenti studi sulla cultivar Leccino, individuano nella selezione FS-17®, un ulteriore e potenziale fonte di resistenza al batterio. Al contrario, si rafforzano le evidenze sull’elevata suscettibilità delle cultivar Cellina di Nardo e Ogliarola salentina. Si presenta infine una rapida sintesi delle diverse attività in corso sulla ricerca di fonti di resistenza.

The biological assay

No abstract

An extensive trial, consisting of overgrafting 12,5 hectares in 3 private olive orchards (located in the infected area of Salento, in Presicce and Ugento) with 2 different levels of symptom's expression, started in April 2016 and completed in the spring 2017 without public funding, is presented. With the collaboration of germplasm collections, scientific institutions and consortia, 260 among different cvs and clones, representative of 19 olive growing countries and all Italian regions, were collected and grafted, by patch grafting, using a complete randomized scheme with 5 replicates and 3 controls on each plants (the susceptible cv Ogliarola salentina and 2 resistant Leccino and Frantoio). Starting from the observation, in the oldest heavily infected area, of almost asymptomatic plants where Leccino were grafted over 10 years ago on trunk of susceptible cvs, the experimental idea has several objectives: try to save the centenary trees by substituting the canopy with resistant varieties; define a grafting protocol adapted to the local conditions; develop a new method "Xylella Quick Tolerance Test" aimed to evaluate the susceptibility of a large quantity of germplasm in shorter time; c. identify a larger number of resistant/tolerant cvs and sources of resistance; d. estimate the potential risk of spreading of the OQD disease in other territories. Due to the good engraftment % and development of grafts the first preliminary evidences are expected in the present growing season.

For Montenegrin vine variety Vranac there is the opinion that high quality wines of this variety can be produced mostly in Mediterranea n climates and there is no possibility of over-spreading of this variety out of warmer areas. The most eastern point of the Vranac cultivation area is the Knjazevac wine-growing region (Serbia), the Potrkanje locality, which is characterized by colder climates (B climate Winkler zone) and an average vineyards altitude of 306 m. lt is in such environmental conditions wine from this variety are regularly assessed in the category of best quality wines "vrhunsko K.G.P.K. vino" with purple stamps of quality and origin within the ,PDO/PGI" system of geographical origin of Serbia. In this paper, a detailed wine analysis was carried out in ltaly using highly sophisticated laboratory equipment , where wine from the Vranac variety from the Knjazevac wine-growing region (Potrkanje locality} from the 2011, 2013 and 2015 vintage was analyzed. Laboratory analysis of quality parameters, including total anthocyanins , intensity of color, color tone of wine (A420, A520 , A620) , etc. for the characterization of the quality and specificity of the wine from the Vranac variety from Knjazevac wine-growing region were performed. In this paper certain positive characteristics of quality of Vranac variety wine are determined , regardless of the fact that the vineyards are in completely different climatic and soil conditions in relation to the originai conditions of Montenegro.

No abstract

While different sources of natural resistance to X. fastidiosa have been described in grapevines and citrus, lack of solid information exists on possible sources of resistance/tolerance in the cultivars that characterize the wide olive germplasm. Preliminary field observations and laboratory analyses of a few cultivars, have shown that differential responses to X. fastidiosainfections exist. To confirm these preliminary findings, a large panel of olive cultivars is being specifically investigated. Currently, the screening procedure relies on field observations looking for symptomless subjects (trees of known cultivars/volunteer seedlings), mechanical inoculations, qualitative and quantitative diagnostic assays (ELISA & qPCR) and, in selected cases, comparative transcriptomic profiling. Field experiments include the planting of the target cultivars/selections in an infected area under high inoculum pressure. All the plots are located in the Apulia Region (Italy) in the demarcated infected area, surrounded by X. fastidiosa heavily affected olive groves. A first experimental plot was established in April 2015 with 10 different cultivars, which was extended in 2016 to 49 cultivars, and will be further enlarged in 2017 with the addition of 40 new accessions. Other plots, comprising newly planted or grafted cultivars (over 260 cvs) have been also established, bringing to over 300 the total number of accessions under evaluation. Cvs Leccino and FS-17®, both expressing interesting traits of resistance, have already been identified.

While different sources of natural resistance to X. fastidiosa have been described ingrapevines and citrus, lack of solid information exists on possible sources of resistance/tolerance inthe cultivars that characterize the wide olive germplasm. Preliminary field observations and laboratoryanalyses of a few cultivars, have shown that differential responses to X. fastidiosa infections exist. To confirm these preliminary findings, a large panel of olive cultivars is being specifically investigated.Currently, the screening procedure relies on field observations looking for symptomless subjects (treesof known cultivars/volunteer seedlings), mechanical inoculations, qualitative and quantitativediagnostic assays (ELISA & qPCR) and, in selected cases, comparative transcriptomic profiling. Fieldexperiments include the planting of the target cultivars/selections in an infected area under highinoculum pressure. All the plots are located in the Apulia Region (Italy) in the demarcated infectedarea, surrounded by X. fastidiosa heavily affected olive groves. A first experimental plot wasestablished in April 2015 with 10 different cultivars, which was extended in 2016 to 49 cultivars, andwill be further enlarged in 2017 with the addition of 40 new accessions. Other plots, comprising newlyplanted or grafted cultivars (over 260 cvs) have been also established, bringing to over 300 the totalnumber of accessions under evaluation. Cvs Leccino and FS-17®, both expressing interesting traits ofresistance, have already been identified.

Notwithstanding the pathogenicy tests on several cvs proved that grapevine is not host of X. fastidiosa. CoDiRO strain, since 2016 hot water treatment (according with EPPO protocol - Bulletin OEPP/EPPO Bulletin, 42(3), 490-492) was adopted as mandatory preventive measures (Commission Implementing Decision (EU) 2015/2417) for all grapevine nurseries in the infected area of Salento. Even if about 10 milions plants have been treated and commercialized without failings in 2016, several market problems occurred for the fear of viticulturists not of Xylella but of possible direct damages on plants. In order to prove the lack of damages a specific trial was started in 2015 comparing not treated material with 3 HWT protocols (the official one 50°C for 45' and 2 more severe 50° 1h and 55° 45') and using 6 different cvs and 5 rootstocks with 75 plants for each thesis. The preliminary results indicate the lack of direct damage in the two HWT thesis at 50° and reveal just a slight delay (about 1 week) in bud breaking. The treatment at 55° cause direct damages to the main buds and a remarkable delay in breaking of secondary buds. After the 2 recent findings of the Pierce disease's agent (X. fastidiosa. subsp. fastidiosa) in Germany and Spain and considering the efficacy of heat treatment against other pathogens and parasites, those results could suggest to spread the adoption of HTW as a safe and common technique in European grapevine nursery industry.

No abstract

In Italy, grapevine certification system requires the status of "virus controlled" for multiplication and commercialization. This system entails free plant propagation material from Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 1, 2, 3 (GLRaV-1, -2, -3), Grapevine fanleaf virus (GFLV), Grapevine fleck virus (GFkV) and Arabis mosaic virus (ArMV)]. Routinely applied bio- and molecular assays for the detection of systemic plant pathogens, such as wood indexing, serological and (RT-) PCR molecular tests, are time-consuming and always require a priori knowledge of the target. On the other hand, the availability of High-Throughput Sequencing (HTS) techniques, which are free from the abovementioned constraints, allows the characterization of the "virome" of the analysed plants. Indeed, data originating from HTS give an unbiased picture of the virus/viroid content of a plant. In this frame, a task of the Apulian Regional Project Re. Ge. Vi. P (Recovery of Apulian grape germplasm) aims to assess the sanitary status of 20 certified autochthonous grapevine clones, table and wine cultivars, by HTS. In 2016, small RNAs (sRNAs) libraries were synthesized from 20 grape accessions and subjected to Illumina sequencing. The downstream data were processed using a custom-developed bioinformatics pipeline including the quality control of the libraries, the de novo assembly of the reads using Velvet assembler and the Blast annotation of the obtained contigs. Annotated contigs confirmed the absence of the above-regulated viruses. Also, this approach showed the presence of two viruses and two viroids not targeted in routinely performed assays, i. e. Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine rupestris vein feathering virus (GRVFV), Hop stunt viroid (HSVd) and Grapevine yellow speckle viroid 1 (GYSVd-1). These results, confirmed by RT-PCR assays, strengthen the potentiality of HTS approach in plant "virome" definition and its application in grapevine certification schemes.

Infoxylella.it is a spontaneous initiative born in February 2016 by a group of researchers, technicians, olive growers, etc., all somehow involved in the Xylella topic, aimed to contrast conspiracy, disinformation and confusion generated, above all through social media, and partly responsible for the opposition and slowdown in the application of containment mandatory measures defined at European level. Infoxylella wants to communicate to and inform transparently public opinion and media with a simple and immediate language. In addition to the website (www.infoxylella.it), the editorial team (more than 30 people), manages a Facebook page and a Twitter profile that in 14 months saw a huge increase in the number of users. The constant commitment and the rigid rules on the dissemination of information only from official and verified sources have progressively increased the credibility and visibility of infoxylella that is often cited or used by journalists and media, like a press agency, as source of information. Although all contents are in Italian, thanks to the online translation utilities, infoxylella has today more than 4800 faithful followers in 45 different countries and a monthly average of 50.000 peoples reached. On behalf of the young experience of infoxylella, we believe that information, together with research results, can play an essential role in the effective application of the containment measures against Xylella and the epidemics in Europe.

No abstract

An intensive movement of varieties along the routes of population's migrations (settlers, traders , pilgrims, sailors and soldiers) occurred since the olden times for Vitis vinifera, one of the most ancient crop spread in central-southern Europe. Recently, the enhancement of varietal identification through DNA markers by genetic profile comparison, lead to the discovery of often unexpected synonyms even in distant regions, thus contributing to gain insight into grape variety circulation in the past. Yet, new questions arise on the movement routes and direction of some ancient grapes, questioning their likely native sites. Although varieties movement took piace also inland (along the Balkan peninsula, in Centrai Europe and across the Alps), most of the spreading of grape cultivars by human trades and migrations occurred in the Mediterranean basin. Here some grape varieties show many synonyms (8-1O and even more) each of them used in a specific area where it was introduced and cultivated, often far from the other locations . Between the two sides of Adriatic Sea, an intensive exchange of materials occurred from/to the ltalian peninsula and the western side of the Balkan area, indicating an intense trade activity in the past. Historical evidences, ampelographic information, pedigree reconstruction and genetic proximity of the involved grapes would ali contribute to shed light on their migration events and/or their possible geographical origin.

The sanitary status of germplasm , together with the genetic aspects , is essential in any clonal selection program and especially when the aim is the recovery and the valorization of minor, neglected and endangered local varieties. Almost always this rare germplasm, often represented by one or few plants escaped from genetic erosion , is infected by single or mixed viruses and should be sanitized in order to evaluate its real quality and technological potential for the wine industry. On the other hand, for the main local varieties such as Vranac or Kratosjia in Montenegro , the virus elimination from interesting candidate clones , could allow to use ali the wide intra-varietal variability identified during the selection, avoiding to lose plants with different and useful characters . The knowledge of sanitary status is important not only for the identification of mother plants free from virus diseases and agents ruled by the legislation on certified grapevine propagation materia! , but also to choose the right technique to eliminate the virus affecting the selections. For the plants infected only by parenchimatic viruses such as the nepoviruses GFLV and ArMV will be used the thermotherapy, for the phloematic viruses (i.e. the GLRaVs, GVA and GVB) the meristem tip culture, for the mixed infection a combination of both techniques. In the framework of the EU funded project EuropeAid /136071/DH/SER/ME "Technical support to renewal of viticulture zoning of Montenegro", 136 accessions selected by 13 jul Plantaze and belonging to over 20 putative local varieties and severa! unknown selections , were tested by ELISA for the presence of 8 different viruses (GFLV, ArMV, GLRaV 1, GLRaV 2, GLRaV 3, GVA, GVB, GFkV), five of them considered in the official EU protocols for the certification of grapevine propagation materials plus 3 additional viruses. In particular, 36 plants resulted negative and the others infected mostly by GVA (38,2% of samp les), GLRaV 3 (32,4%), GLRaV 1 (31,6%), GFLV (22,1%), GFkV (11,8%) and GVB (3,7%). The results, compared with the situationin otherMediterraneanareas with similar geographic characteristics , showed a relatively good sanitary status reflecting the low presence of natura! vectors and probably also the isolation of native germplasm in some restricted marginai areas. The 36 plants negative in ELISA were re-tested by the most sensible RT-PCR (adding also Grapevine Pinot Gris Virus not yet reported in Montenegro) resulting in 7 samples infected just with GFLV and 29 samples (21,3%) confirmed as negative to ali viruses tested . On behalf of some success story of valorization of neglected varieties for example in Apulia region, those preliminary data , hopefully with the further steps such the sanitation of some interesting selections , the graft indexing and the evaluation of candidate clones in comparison field, could enrich in the future the number of Montenegrin local varieties in the national catalogue and the list of unique and new quality wines on the market.

Vegetative propagation of grapevines favours the perpetuation and dissemination in nature, of intracellular pathogens (over than 70 considering viruses, viroids and phytoplasmas). Once in the field, their presence may affect in various ways the plant's yield and its quality. Preventing infection spreading through a prompt identification, characterization and exclusion approach, is still the most effective action of containment. To this aim, in 2014, registered native grapevine cultivars from Apulia were analyzed to determine their "absolute" sanitary status (virome), i.e. the totality of the infectious agents present in any single accession using a high throughput sequencing (HTS) approach. Small RNA libraries were therefore synthesized and analyzed for the presence virus/viroid sequences. Among these, a symptomless accession D205 of cv. "Bombino nero" growing in a foundation block showed the presence of 21 contigs (ranging from 329 to 56 nt in length) identified as badnavirus-like. Using the closest badnavirus found by BLASTX search as a reference sequence, i.e. Fig Badnavirus 1 (FBV-1, accession No. NC017830), a set of specific primers was designed. Amplification with these primers produced the expected amplicon, which has 91% nucleotide similarity identity with a badnavirus recently discovered in Greece and denoted Grapevine Roditis leaf discoloration-associated virus (GRLDaV). For a preliminary assessment of the incidence of this virus in the field, a preliminary survey in the same foundation block was conducted. A total of 11 samples from different autochthonous cultivars were checked by PCR but none of them tested positive.

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