Field symptoms of chlorotic mottling and leaf deformations were observed on the cv Pinot gris (PG) in the Trentino region (Italy). Extensive assays excluded the presence of widely distributed nepo-, ampelo- and vitiviruses. An analysis of small RNA populations from two PG grapevines showing or not symptoms was carried out by Illumina high throughput sequencing. The study disclosed the virus and viroids contents of the two vines that was composed by Grapevine rupestris stem pitting-associated virus (GRSPaV), two viroids Hop stunt viroid (HSVd) and Grapevine yellow speckle viroid 1 (GYSVd1), the marafiviruses Grapevine rupestris vein feathering virus (GRVFV) and Grapevine Syrah virus 1 (GSyV-1), and a hitherto unrecorded virus. This virus had a genome organization identical to that of Grapevine berry inner necrosis virus (GINV), a trichovirus reported only from Japan, with which it grouped in phylogenetic trees constructed with sequences of the RdRp domain and the coat protein gene. However, molecular differences with GINV are wide enough to warrant classification of the virus in question as a new species, for which the provisional name of Grapevine Pinot gris virus (GPGV) is proposed. A limited field survey for the presence of GPGV in diseased and symptomless plants from three different cultivars did not allow to clearly associating the virus to the observed symptoms. © 2011 Elsevier B.V.

The paper reports the results of a project of recovery and molecular characterization aimed at identifying the local vine varieties cultivated on the archipelago of Malta. A total of 58 accessions from the two major islands, Malta and Gozo, were sampled within the framework of a clonal and sanitary selection program. DNA analyses were conducted using 13 SSR markers and the profiles obtained compared with the CRA-VIT archive. A total of 28 different genotypes were identified, eight of which correspond to imported varieties: Ansonica, Axina de tres bias, Calabrese, Gold, Koenigin der weingaerten, Muscat fleur d'oranger, Muscat Hamburg, and Negroamaro. Reference SSR profiles have been produced for the most important local varieties, Girgentina (Insolja tal-Girgenti), Gellewza, and Gennarua (Insolja ta' Gennaru).

Analysis of aerial photos by using GIS tools and on-field surveys of flora are used to characterize territories from an agro-ecologica I point of view and to assess the level of diversity of given agro-ecosystems. More and more correlations between landscape characteristics, sustainability and quality of agriculture production were speculated. In last three years a study was carri ed out in the area of DOC "Gioia del Colle" in Apulia, South Italy, in order to characterize and investigate different vineyards and sites and find out possible interactions and correlations between the landscape diversity, the biodiversity of fields and the quality of grapes and wines. In order to investigate such aspects and achieve a better knowledge of such correlations, 13 vineyards were continuously monitored for qualitative aspects related to grapevine phenology, growing and cultivation as well as for investigating quality of flora and biodiversity of spontaneous plants. A landscape analysis was carried out identifying a buffer area of a radium of 5 km around each of the vineyards and by carrying on the calculation of a set of indicators able to quantify landscape structure, composition and level of connection on the basis of the different shape, areas and cover of the patches. Statistical correlations between values of the landscape indicators, biodiversity of spontaneous plants of the different representative vineyards and the specific characteristics of the grapes were analyzed. A preliminary better knowledge of the interactions between such elements of the landscape, the sustainability of grapevine cultivation and the quality of the grapes was achieved.

Two dsRNA molecules with an estimated lenght of 1.5 Kbp were identified and characterized from leaves of a Japanese persimmon (Diospyros kaki) tree, showing veinlets necrosis on both sides of leaf blades.DOP-PCR recognized two genomic fragments of a bipartite cryptic virus, for which the name of Persimmon cryptic virus (PeCV) is proposed. RLM-RACE leaded to the sequencing of a 1,510 bp contig identified as dsRNA-1 and a 1,491 bp complete segment identified as dsRNA-2. The two genomic fragments resulted both monocistronic and harbored conserved domains related to RNA-dependent RNA polymerase (RdRP) and capsid protein (CP) of species associated to Alphacryptovirus genus. Phylogenetic analysis of RdRP sequence showed highest amino acid identity with Black raspberry cryptic virus (BrCV, 63 %), Pepper cryptic virus 2 (PCV-2, 52 %) and -1 (PCV-1, 46 %). Whereas the CP putatively encoded by dsRNA-2 shared highest identity with Mulberry cryptic virus 1 (MCV-1, 49 %), PCV-2 (39 %) and PCV-1 (32 %). N-J phylogenetic analysis confirmed those relationships and delivered PeCV in a cluster with phyto-cryptoviruses belonging to genera Alpha- and Betacryptovirus, quite far distinguished from myco-cryptoviruses, gathered in genus Partitivirus.Virus-specific primers for RT-PCR were successfully designed inside the CP region to detect PeCV in several symptomless trees found in different orchards of Apulia (Southern Italy), thus proving that infection may be fairly common and presumably latent.Policlonal antiserum (kindly provided by Dr. M. Turina, CNR-IVV, Torino, Italy) specific to the CP of family-related Beet cryptic virus 2 (BCV-2) was profitably used for western blot detection of a 45-50 KDa band, coherent with predicted size of dsRNA-2 product. Furthermore, antibodies were useful for ISEM observation and subsequent decoration of PeCV particles, proved to be isometric, around 30 nm in diameter, with rounded shape lacking in fine structural details, not easily permeable by negative stain.

Grapevine rupestris stem pitting-associated virus (GRSPaV, genus Foveavirus, family Betaflexiviridae) is the putative agent of the Rupestris stem pitting (RSP) disease of grapevines. GRSPaV comprises a family of variants whose pathological characteristics are incompletely known. Recently, many of the autochthonous table grape cultivars were tested for the presence of GRSPaV in two major Turkish grape-growing areas, i.e. the Eastern Mediterranean and the Southeast Anatolia regions. Comparative analysis of local GRSPaV isolates from these native cultivars was performed with viral genome sequences from NCBI database. To our knowledge, this is the first report for the presence of GRSPaV in Turkish vineyards.

Through the application of next generation sequencing, in synergy with conventional cloning of DOPPCR fragments, two double-stranded RNA (dsRNA) molecules of about 1.5 kbp in size were isolated from leaf tissue of a Japanese persimmon (accession SSPI) from Apulia (southern Italy) showing veinlets necrosis. Highthroughput sequencing allowed whole genome sequence assembly, yielding a 1,577 and a 1,491 bp contigs identified as dsRNA-1 and dsRNA-2 of a previously undescribed virus, provisionally named as Persimmon cryptic virus (PeCV). In silico analysis showed that both dsRNA fragments were monocistronic and comprised the RNAdependent RNA polymerase (RdRp) and the capsid protein (CP) genes, respectively. Phylogenetic reconstruction revealed a close relationship of these dsRNAs with those of cryptoviruses described in woody and herbaceous hosts, recently gathered in genus Deltapartitivirus . Virus-specific primers for RT-PCR, designed in the CP cistron, detected viral RNAs also in symptomless persimmon trees sampled from the same geographical area of SSPI, thus proving that PeCV infection may be fairly common and presumably latent.

Extensive necrosis of the veinlets were observed in early summer of 2011 on both sides of the leaf blades of a Japanese persimmon (Diospyros kaki) tree growing in a private garden in the vicinity of Bari (Apulia, southern Italy). This tree (accession SSPI) hosted a previously uncharacterized cryptovirus whose presence was apparently unrelated to the vein necrosis condition as it occurred also in a large number of symptomless trees (Morelli et al., 2012). Highthroughput sequencing analysis of the SSPI virome, performed on dsRNA extracts with an Illumina platform, allowed the identification of four contigs which, upon BLAST analysis (Altschul et al., 1997) showed a 97% sequence identity at the nucleotide level with sequences of the putative cytorhabdovirus Persimmon virus A (PeVA, GenBank AB735628), recently described in Japan (Ito et al., 2013). The sequence of the largest contig (983 bp), spanning part of the polymerase gene and the 3'-UTR, was deposited in GenBank under the accession No. KM407515. To confirm deep-sequencing findings, the PCR primer set PeVAfor/Pe- VArev (5'-AGGATCATTACAAAATCCGTGAGG-3'/ 5'- TTCCCGAAAGACAATCTGTCCC-3'), intended to amplify a 250 bp product, was designed on the KM407515 sequence. An amplicon of the expected size was repeatedly obtained from the symptomatic (SSPI) but not from 10 symptomless trees. This product was cloned into pSC-A-amp/kan and custom-sequenced (Macrogen Europe, The Netherlands). BLAST analysis matched previous identification, as cloned sequences shared ca. 98% identity with those of the Japanese PeVA isolate. There was no amplification when RT-PCR assays were extended to the 10 symptomless persimmon trees. Whether or not PeVA is involved in the induction of vein necrosis remains to be ascertained. To our knowledge, this is the first report of PeVA in a country other than Japan.

Forty-five grape accessions, traditional and historically mentioned in Apulia (southeastern Italy), were genotyped at 13 microsatellite (SSR) markers and observed for their morphological features with the aim of characterizing and identifying the local grape diversity relevant for economic or historical significance and for endangered germplasm conservation. Twelve of the 45 accessions examined were found to be synonyms or somatic mutants, leaving 33 distinct genotypes. Attempts were then made to verify the true identities of the accessions investigated and to determine their appropriate denominations. This entailed comparing them with published allelic profiles and morphological features of cultivars from Apulia and from surrounding areas linked historically to the region. While confirming the identity of the major Apulian cultivars, further matches with varieties from other Mediterranean regions were revealed. Approximately half of the Apulian cultivars investigated were found to have a foreign counterpart mainly along the Adriatic Sea (Croatia), in Greece, or in other southern Italian regions. The new synonymies found with cultivars traditional to other areas shed light on the migration of cultivars following the settlement of colonies and the historical establishment of Mediterranean trade routes. © 2014 by the American Society for Enology and Viticulture. All rights reserved.

Nell"ambito del Piano di sviluppo rurale 2007-2013, la regione Puglia ha identificato nella misura 214/4 sub-azione a): "Progetti integrati per la biodiversità" l"importanza di tutelare, caratterizzare e identificare il numeroso germoplasma autoctono regionale. La regione presenta un territorio molto vasto e ricco di germoplasma autoctono vegetale, che corre il rischio di essere soggetto ad una graduale erosione genetica e quindi ad una perdita di biodiversità irreversibile. L"obiettivo principale dei progetti Re.Ge.Vi.P., Re.Ger.O.P., e Re.Ge.Fru.P., occupandosi rispettivamente di vite, olivo e fruttiferi, è di realizzare, attraverso un approccio integrato multidisciplinare, un effettivo recupero e studio del germoplasma propedeutico alla valorizzazione della agrobiodiversità regionale. La prima fase del lavoro consiste in una minuziosa ricerca bibliografica in fonti edite e inedite, ma anche attraverso le testimonianze e la memoria storica degli agricoltori su usi e tradizioni concernenti le vecchie varietà coltivate. La ricerca, l"individuazione sul territorio e georeferenziazione del germoplasma stanno consentendo di recuperare il materiale arricchendo le collezioni ex situ già esistenti. È inoltre previsto che tutto il materiale recuperato sia caratterizzato mediante approcci morfologici, molecolari, agronomici tecnologici e sanitari al fine di verificare l"identità varietale, chiarire l"esistenza di omonimie e sinonimie, scoprire nuova biodiversità intra-varietale, registrare ufficialmente le nuove accessioni di germoplasma nonché valutarne le potenzialità economico-produttive per una possibile ripresa in coltivazione. Nell"ambito dei progetti, oltre a costituire delle collezioni ex situ di riferimento per l"intera regione, secondo le linee guida rilasciate nel 2012 dal Gruppo di lavoro per le "Risorse genetiche vegetali" del MiPAAF, l"inserimento del germoplasma in collezioni private e presso aziende custodi consentirà una conservazione bilanciata tra attività in situ ed ex situ. Infine tutte le nuove informazioni acquisite, catalogate in un archivio elettronico fruibile al pubblico, costituiranno al contempo un dettagliato strumento di conoscenza e divulgazione dell"enorme patrimonio viticolo, olivicolo e frutticolo regionale. Nel prossimo programma di sviluppo rurale pugliese, si auspica che i risultati dei progetti integrati, insieme alla recente legge regionale di "Tutela delle risorse genetiche autoctone" (L.R. n.39 dell"11/12/2013), possano concretizzarsi in reali iniziative di valorizzazione del germoplasma locale.

The old world and the new world wine-making philosophies tried to explain wine quality with role of nature and organoleptic characteristics of the wine. Among the various environmental factors specifically, c1imate and soil are the most important factor on viticulture zoning due to its direct effect on wine development and wine quality. The implementation at locallevel of GIS (Geographic Information System) could play a key factor to define some parameters in order to create homogenous zones. Main objective of the work is create a thematic database with a GIS implementation that can have some innovative aspects based on geomorphological, climatic, soil and land use ofthe area. GIS is able to analyse, design and create a set of descriptive indicators ofterritories characteristics, using spatial interpolations to create specific maps of studi ed area. GIS tool some other data can be compared such as the immediate characteristics of grapes, in order to understand the best suitable areas for the production of quality wines. According to data, the GIS tool has defined some parameters based on geomorphology and pedology, c1imate and other more specific measurements such as Winkler index, Huglin index and Evapotranspiration. As a result it allow us to get a computerized recording ofthe zoning, shows the crossing of different levels of information, such as texture, and c1imate and some other information. The study has been carried out in Italy, in Puglia Region. In the AOC "Gioia del Colle" on Primitivo variety. Results of the GIS tool, shows that some areas of the denomination of ori gin can bave similar characteristics in terms of climate values, soil quality and cultivar characteristics. These information have been confirmed from Winkler index, Huglin index and Evapotranspiration measurements .. Thirty-two fields have been selected according to density and age and some chemical and physical parameters have been analysed in 2013 in order to characterize their poliphenolic composition.

To investigate the role of herbaceous plants as hosts of Xylella fastidiosa, monthly sampling of the native flora of two heavily infected olive groves and of the sides of the adjacent dirt roads was conducted from January 2014 onwards. One of the orchards had not been subjected to weeding, whereas periodic tillage had been carried out in the other. Overall, more than 100 species of 40 monocotyledonous and dicotyledonous families were collected.

To investigate the role of herbaceous plants as hosts of Xylella fastidiosa, monthly sampling of the native flora of two heavily infected olive groves and of the side of the adjacent dirt roads was conductedfrom January 2014 onwards. One of the orchards had not been subjected to weeding, whereas periodic tillage had been carried out in the other. Overall, more than 100 species of 40 monocotyledonous anddicotyledonous families were collected, photographed and identified, their phenological stage was recorded using the Keller-Baggiolini scale, their period of presence and the type of distribution prevailing in the field (e.g. whether the species scattered or concentrated under olive trees or along the edges of the dry stone walls) was assessed, as well as the abundance-dominance indices according to the Braun-Blanquet method. At all sites monitored, Philenus spumarius, the main and most important vector of Xylella fastidiosa so far found in the area of the outbreak, was present with abundant populations on herbaceous species in the spring and on olive trees from the middle of May throughout the summer. All weed samples collected, in pools of 4-5 plants, were tested by DAS-ELISA and the uncertain results were verified by PCR. So far, none of the samples analyzed, in excess of 600, proved to host Xylella fastidiosa, confirming the preliminary observations that, by and large, weeds may not have a major role in the epidemiology of X. fastidiosa in the considered area. However, since sampling of the summer/autumn flora is yet to be done, a better insight into the epidemiological role of these plants will become available in the coming months, in conjunction with the migration of the vectors from the olive trees onto the native flora with the consequent possible inoculation of the bacterium.

The presence of eight viruses (Arabis mosaic virus ArMV, Grapevine fanleaf virus GFLV, Grapevine fleck virus GFkV, Grapevine leafroll-associated viruses 1, 2 and 3 - GLRaV-1, -2, and -3, Grapevine virus A GVA, Grapevine virus B GVB) was tested using ELISA in a collection of woody indicator vines located at the Faculty of Agriculture Zagreb. Out of 75 tested plants (15 from each indicator - LN33, 110 Richter, Vitis riparia Gloire de Montpellier, V. rupestris St. George and Kober 5BB) six of them (one of 110R and V. rupestris, two of LN33 and Kober 5BB) were positive to ArMV, one V. rupestris to GLRaV-1 and one V. riparia to GLRaV-2. All the infected plants were cut off and their roots killed using glyphosate.

The agricultural biodiversity, in addition to the need of protecting the genetic and historical-cultural heritage, is a significant untapped economic potential that, albeit slowly, was started to be used. Its contribution to the local agricultural economy can be declined in several ways: the growing demand for new products strictly territorial in response to the product's standardization on the global markets; the placement of biodiverse products in new marketing channels such as the direct sale or the direct consumption in the agritourisms; the use of more adaptable local cultivars in marginal areas with lower costs and energy inputs. The agro-biodiversity, in the logic of the diversification of the agricultural activities, is being re-evaluated especially in the small multifunctional farms interested in the receptivity and valorisation of the territory. The same in-situ and ex-situ germplasm collections represent a further important tourist attraction to be exploited for increasing, especially out of season, the flow of food and wine enthusiasts in the rural areas. The Apulia Region, through the EU Rural Development Plan (RDP 2007-2013), decided to funds 5 "Integrated projects for biodiversity" aimed to identify, collect, protect and characterize the rich germplasm of the main crops represented by hundreds of old/minor/neglected cvs threatened by genetic erosion just before their irreversible loss. With big multidisciplinary efforts, the projects Re.Ge.Vi.P., Re.Ger.OP and Re.Ge.Fru.P., dealing respectively with grape, olive and fruits crops, allowed to make a sensible progress in the recovery and study of regional germplasm representing the prerequisite for its economic valorisation. The wide partnerships, through 8 activities, including bibliographic research, meetings and interview with the farmers, identified, geo-referenced, collected, multiplied and planted several hundred new selections/varieties, enriching the existing ex-situ gene banks; all the germplasm was studied and characterized by morphological, genetic, agronomic, technological and sanitary approaches in order to verify the varietal identity, to register officially the new varieties as well to evaluate its real economic and productive potential. The field collections with improved facilities/labs, the new knowledge acquired in open data-bases, the new regional law "Protection of indigenous genetic resources" and the RDP funds 2014-2020, all together represent the necessary elements of our long-term germplasm valorization' strategy.

E' stata recentemente decretata (D.M. 22 aprile 2011) la registrazione al Catalogo nazionale delle varietà di vite di 5 vitigni autoctoni pugliesi ad uva da vino (Antinello B., Marchione B., Maresco B., Minutolo B. e Somarello Rosso N.), atto conclusivo di un lungo lavoro svolto dai ricercatori pugliesi e punto di partenza per la successiva valorizzazione commerciale. Le attività che hanno portato al riconoscimento ufficiale comprendono il recupero del germoplasma, la caratterizzazione ampelografica e molecolare, il miglioramento sanitario attraverso risanamento, il confronto e la determinazione delle caratteristiche produttive ed enologiche secondo i protocolli codificati dalla normativa. I 5 vitigni, fino ad ora coltivati marginalmente in vigneti promiscui, hanno così acquisito identità propria e guadagnato la possibilità di indicare il nome in etichetta, uscendo dall'anonimato di un uso localistico/amatoriale per proiettarsi in una dimensione produttiva regionale. Per Minutolo e Somarello rosso, in particolare, la registrazione dei primi tre cloni potrà contribuire al miglioramento delle condizioni sanitarie e quindi della redditività dei nuovi vigneti specializzati. Attraverso brevi note storiche e la descrizione delle caratteristiche produttive/tecnologiche, vengono presentate le varietà, esempi concreti di un più ampio lavoro sulla biodiversità agraria regionale nonché nuovi strumenti al servizio del comparto enologico pugliese: Maresco e Maruggio idonei, per l'elevata acidità dei mosti, alla produzioni di vini base spumante; Somarello rosso per vini rosati monovarietali; il Minutolo, con intensi aromi di Moscato, già impiegato, con 25 etichette sul mercato, per vini bianchi di alta qualità, passiti e spumanti; infine l'Antinello per caratterizzare geograficamente vini bianchi in uvaggio con altri vitigni locali.

The Knjazevac wine-growing region is a typical B (WIN) viticulture zone with warm (HI) and humid (DI) climate classes. With higher elevation terrains, characteristics of the relief between the Timok River valley and surrounding mountains, vertisol as type of soil, as well as CI + 2 Cold nights index etc., the quality and characteristics of wine in this region are greatly influenced by some parameters of Terroir. Wine producers from this wine-growing region recognized these specificities, founded a representative association and registered the protected designation of origin (PDO) in the new Serbian PDO wine system, established and harmonized to the EU legislation through the results of the Twinning Project SR08IBAG02 ,,Capacity building and technical support for the renewal of viticulture zoning and for the system of designation for wine with geographical indications in Serbia" (2011/2013). This paper presents the analysis of some of the main ecological and anthropogenic factors of the Knja?evac wine-growing region which greatly effect wines with a medium to high alcohol content which have, at the same time, a pronounced lightness and freshness due to a medium-high acid content. In particular, the focus was on some factors analyzed in several representative locations/vineyards such as altitude, exposition, slope, varieties, rootstocks, vineyard surface, training system, etc. which certainly represent the Terroir of the Knja?evac wine-growing region and have a big impact on the quality and characteristics of the PDO wines.

Five viroid species have been reported fromgrapevine. Hop stunt viroid (HSVd) and Grapevine yellowspeckle viroid 1 (GYSVd-1) are distributed worldwide,whereas Grapevine yellow speckle viroid 2(GYSVd-2), Australian grapevine viroid (AGVd) andCitrus exocortis viroid (CEVd) are found only sporadically.However, the presence of AGVd and GYSVd-2 inseveral countries, including China, Turkey and Tunisia,suggests a wider dissemination, possibly also in Europe,where AGVd has never been found and GYSVd-2 hasbeen occasionally identified in Italy. Taking advantage ofa multiplex RT-PCR assay recently developed for detectingsimultaneously these five viroids, vines growing inItaly in commercial vineyards and germplasm collectionswere surveyed. Besides confirming the widespreadpresence of HSVd and GYSVd-1 in the field, GYSVd-2and/or AGVd were identified in two grapevine tablecultivars (Sultanina Bianca and Red Globe) from germplasmcollections. Tests extended to vines cultivated insouthern Italy confirmed the presence of both viroids,which were further characterized. No major sequencedivergences between the AGVd and GYSVd-2 variantsfrom Italy and those previously described from othercountries were observed. Phylogenetic analysis supportedthe close relationships among AGVd variants from Italy,Tunisia and Australia. To our knowledge this is the firstreport of AGVd in Europe and the first molecular characterizationof GYSVd-2 isolates from a European country.

The Directive 2009/128/EC on the sustainable use of pesticides aims at promoting the use of Integrated Pest Management and alternative approaches, such as the selection of new germplasm more tolerant or resistant to pests and diseases. In the frame of collaboration with Balkan breeders, two experimental fields were planted with 27 new cultivars obtained by crosses between Vitis vinifera (L.) and other species of the genus Vitis, and six reference cultivars commonly grown in Italy (Italia, Red Globe, Victoria, Michele Palieri, Black Magic and Crimson Seedless). In particular, nine cultivars were supplied by the Research Institute for Viticulture and Enology, Pécs (H), seven by the National Institute of Winegrowing and Winemaking, Chisinau (MO), and 11 by the Experimental Station "Sremski Karlovci" of the Institute for Pomology and Viticulture, University of Novi Sad (SE). Two years of field observations and in vitro assays were carried out to evaluate the adaptability of the new germplasm to the pedo-climatic conditions of Apulia (southern Italy). All the new cultivars showed an overall lower susceptibility to downy mildew as compared to the reference ones in the field as well as in vitro. Downy mildew infections were limited and sporification sparse and concentrated at the edges of the infected area. The most promising were the Hungarian cvs Eszter, Palatine, Bolgar Rezy, Terez, and the Serbian cvs Poleskey Muskotaly, Venus, and Piroska. An in vitro assays showed that cvs Palatine, Poleskey and Muskotaly were also more tolerant to powdery mildew as compared with the other cultivars.