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INTRODUCTION – Common limits for phenotyping are costs in terms of equipment, specialized researchers, and time. Moreover, phenotyping requires specific methods in relation to the studied traits, and each method is generally designed to highlight very specific characteristics of the singular trait. Thus, the obtained results are not easily comparable between different institutions. However the data sharing is clearly the most powerful approach to limit the costs and efforts optimizing the obtained results in term of number of studied accessions. It should be stressed that funds are another great limit in research, especially in East countries, where the most interesting cultivar variability is preserved. 28 AIMS AND SCOPES – The main purpose of this work is the establishment of a phenotyping network able to produce comparable data among the Pan-European grapevine collections. MATERIALS AND METHODS – Two protocols were proposed: one for phenology and, the other, for enocarpological traits. Data were recorded in two consecutive vintages: 2012 and 2013. Concerning phenology, the extended BBCH scale (universal and flexible) was adapted to our objects by moving it into a chrono- phenological scale. Then, a cubic curve was selected for modelling the phenological courses. Thus, computing for each accession specific coefficients, it was possible to obtain the relative phenograms. Considering the different site climatic conditions, it was possible to spatialize the results, obtaining reference phenological maps. Concerning eno- carpological traits, the attention was focused on characteristics with a clear impact on the production quality, such as berry morphology (weight, diameters, number of seeds …) and composition (sugar, titratable acidity, anthocyanin and phenolic contents …). The protocol was organized to concatenate the data acquisition, with the objective to generate new derived variables. The methods were selected to be easily reproduced in all the involved collections, without the necessity of expensive facilities. RESULTS AND DISCUSSIONS – The first year of dataset was used to evaluate the methods. Two publications are already available confirming the protocols quality. Concerning phenology, both the variability between accessions grown into the same collection and the phenological shifts between the reference varieties cultivated in different areas were highlighted by the protocol. Matching climatic and phenological information of reference cultivars, phenophases maps were obtained. The performances obtained by the eno-carpological protocol were satisfactory both in a global point of view and in a singular collection evaluation. The project obtained an impressive participation, considering the volunteer contribution without any material funding to cover the work costs. Twenty-four institutions collaborate to the data collection. 1724 accessions were evaluated: 931 (583 in 2012 and 348 in 2013) for phenology and 793 (424 in 2012 and 369 in 2013) for eno-carpological descriptions. CONCLUSIONS AND POSSIBLE APPLICATIONS – The protocol sharing allows the data comparison among different grapevine collections. The possible future application for this dataset could be related to scientific studies of genetic associations. Moreover, the improved knowledge of minor and neglected cultivars could have a direct impact on the industrial production, giving information concerning new varieties for future introduction in viticultural areas.

INTRODUCTION – Cryopreservation is a method for long-term conservation in which explants are stored at the ultra-low temperature of liquid nitrogen. In the case of grapevine, cryopreservation protocols have not been defined enough for wide application to numerous cultivars. Cryotherapy is a novel technique, which eliminates plant pathogens during exposure to ultra low temperatures. Plant cryopreservation is a complex procedure during which explants are exposed to potentially stressing events. Genetic stability of regenenerated plants is a part of cryopreservation protocol establishment. AIMS AND SCOPES – The main goals of this research are: to determine the efficient cryopreservation protocol for Croatian native grapevine cultivars and international cultivars, to test efficiency of cryotherapy in virus eradication and to check genetic stability of regenerated plants. This research was performed aiming to introduce this technique in general viticulture. MATERIALS AND METHODS – In vitro explants of Croatian (Plavac mali, Maraština-(plus their infected genotypes), Pošip and Škrlet) and international cultivars (Chardonnay, Cabernet Sauvignon, Merlot and Pinot Noir, within their infected genotypes) were taken for cryopreservation experiment, which had been established previously on cultivar Portan. Droplet- vitrification protocol was performed. Regenerated two-months in vitro plants from infected genotypes (control and cryopreserved apices) were subjected to ELISA testing. AFLP markers were employed to test genetic stability on cultivar Portan. Eight AFLP primer combinations were used on cryopreserved and non-cryopreserved shoot tips (sampled after sucrose preculture, loading and treatment with half-strength PVS2 solution) for a total of 43 plants tested. RESULTS AND DISCUSSIONS –The basal cryopreservation protocol, performed on international cultivars revealed: intermediate regrowth was achieved with Chardonnay (30%) and high with Merlot (70%), before and after cryopreservation, which was comparable to previously published results on 10 Vitis cultivars, which produced an average recovery of 64%. Similar results were obtained with healthy and virus infected cultivars. The same protocol, performed on Croatian cultivars, showed a low survival and recovery with only one (Maraština) of the four cultivars tested. This low success might be due to the fact that cryopreservation experiments were performed using cultures which had been introduced in vitro very recently and that the in vitro culture conditions employed were not adapted to the multiplication of these Croatian cultivars. The results of the cryotherapy experiments revealed that a high proportion of Chardonnay (100%) and Cabernet Sauvignon (78%) plants were virus-free after being subjected to cryopreservation. Control plants showed a high proportion of healthy plants, almost the same as with LN exposed ones for both viruses. AFLP markers revealed no significant differences in AFLP profiles after sucrose preculture, treatment with the loading solution and with half-strength PVS2, whereas an increase in polymorphic fragments was observed in non- cryopreserved and cryopreserved samples treated with PVS2 solution, the number of polymorphic fragments increasing with longer durations of exposure to PVS2 solution. CONCLUSIONS AND POSSIBLE APPLICATIONS – Cryotherapy as a method can be efficient for virus eradication even in the preconditioning steps of a cryopreservation protocol. A genetic stability study should be included in post- cryopreservation experiments.

INTRODUCTION – Before Philloxera crisis in Croatia were more than 400 grape varieties under cultivation, but today their number is drastically decreased. Recently, many efforts in the preservation of grapevine biodiversity have been undertaken: detailed inventory of wine-growing regions, establishing of National and regional collections and their systematic ampelographic and genetic characterisation. Recent genetic research resulted in revealing synonyms and homonyms, as well as genetic relationship within Croatian native cultivars, as well as with cultivars from other regions. Evaluation of quality traits of native cultivars has also started. AIMS AND SCOPES – The main goals of this research are: to determine the number of Croatian native grapevine cultivars based on inventarisation of viticultural areas, ampelographic and genetic characterization ; to perform detailed ampelographic characterization using OIV descriptors ; to perform the genetic characterization of Croatian native grapevine cultivars using SSR markers and to determine the level of genetic variability within Croatian native grapevine cultivars. MATERIALS AND METHODS – Vines of a potentially native cultivars ware selected and marked and used as a source of propagating material for establishment of a National collection of Croatian grapevine cultivars located at experimental station Jazbina, at Faculty of Agriculture, University of Zagreb. In this process more than 140 accessions were introduced to this collection. Phenotyping of collected accessions was performed using 47 OIV descriptors for grapevine cultivars and Vitis species that were selected within the GrapeGen 06 and used for cultivars description in European Vitis database. SSR genotyping was performed using a total of nine markers. Six SSR markers (VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79) recommended by OIV and three additional ones (VVMD25, VVMD28, VVMD32) selected during the Grapegen06 project. RESULTS AND DISCUSSIONS – Based on results of inventarisation, ampelographic characterization and genetic characterization the list of 103 Croatian native grapevine cultivars is defined together with their synonyms and population status/vulnerability evaluation. Based on presented information on population status of Croatian native grapevine cultivars we can conclude that more than 50% of cultivars are critically endangered, endangered or vulnerable. The results of ampelographic characterization of Croatian native cultivars are showing the high level of variability. Comparison of Croatian cultivars SSR profiles with the results of the other cultivars in EU- vitis database confirmed that majority of them are unique and exist only in Croatia. Since most of them are duplicated in grapevine germplasm collections throughout Croatia they are considered as “saved”. Ongoing genetic studies with increased number of SSR loci (>30) as well as use of SNP markers will allow us more detail analysis of genetic structure and relatedness of Croatian native cultivars. CONCLUSIONS AND POSSIBLE APPLICATIONS – The preserved native cultivars are presenting a good base for future development of Croatian viticulture and because of high genetic variability, as well as some traits that are detected within Croatian native cultivars they could be of interest to other countries for production as well as for breeding.