‘Žilavka’ has been grown in Bosnia and Herzegovina since the XIVth century and is exploited for wine production. Although not sufficiently studied, this grapevine cultivar has high economic potential ...for the country. Five survey missions resulted in the collection of eighty ‘Žilavka’ accessions that varied in terms of names and characteristics. Because of the unknown origin of the cultivar, these accessions were analyzed with microsatellites markers in order to obtain a standard ‘Žilavka’ genotype. AFLP markers were used to investigate the genetic basis of variability within the cultivar. ‘Žilavka’ grapevines were screened on 14 microsatellite loci, thus revealing 4 different genotypes arising from mutations observed at 10 polymorphic loci. AFLP analysis of 52 ‘Žilavka’ accessions revealed 35 different genotypes, with an average polymorphism of 57 %. Cluster analysis showed no grouping of different Žilavka accessions according to their names, characteristics or collection locations. The standard ‘Žilavka’ genotype was further compared to 211 cultivars from Slovenia (49), Austria/Germany (20), France (13), Portugal (27), Croatia (19), Greece (32), Spain (21) and Italy (30) in order to assess their genetic relationships. In pairwise comparisons, the highest genetic similarity was found with Slovenian cultivars ‘Glera’ and ‘Briška Glera’ (64 %) and the highest genetic dissimilarity (100 %) with two Italian cultivars, ‘Nebbiolo Lampia’ and ‘Vespolina’. Inventory, collection and genetic characterization of ‘Žilavka’ accessions are important steps towards cultivar standardization, identification of parental cultivars and investigation of cultivar origin, required for its sustainable use.
‘Žilavka’ je pomembna vinska sorta z območja Bosne in Hercegovine kjer jo gojijo že od XIV stoletja. V preteklosti ni bila dovolj proučevana vendar ima velik ekonomski potencial za omenjeno območje. Akcesije žilavke (80), ki imajo različna imena in se razlikujejo po nekaterih osnovnih karakteristikah so bile nabrane na različnih področjih Bosne in Hercegovine. Za pridobitev standardnega genotipa žilavke smo uporabili mikrosatelitne markerje in z analizo 14 lokusov odkrili mutacije na 10 lokusih in določili 4 različne genotipe. Vse akcesije (52) katerim smo na osnovi mikrosatelitov določili standardni genotip smo nadalje analizirali z AFLP markerji in tako določili 35 različnih genotipov. Povprečni polimorfizem pri AFLP analizi je bil 57 %. S klastrsko analizo nismo odkrili skupin povezanih z različnimi karakteristikami, poimenovanjem ali izvorom akcesij žilavke. Standardni genotip žilavke smo nadalje primerjali z genotipi 211 kultivarjev iz Slovenije (49), Avstrije/Nemčije (20), Francije (13), Portugalske (27), Hrvaške (19), Grčije (32), Španije (21) in Italije (30), da bi ugotovili kakšna je sorodnost z njimi. V analizi primerjave parov smo ugotovili, da je žilavka najbolj sorodna (64 %) s slovenskima kultivarjema ‘Glera’ in ‘Briška Glera’ ter najbolj genetsko oddaljena od italijanskih kultivarjev ‘Nebbiolo Lampia’ in ‘Vespolina’. Zbiranje, vrednotenje in genetska karakterizacija akcesij žilavke je pomembna za standardizacijo kultivarjev, analizo starševstva in izvora sorte.
To introduce testing of Grapevine leafroll-associated virus-2 (GLRaV-2) and Grapevine virus B (GVB) in sanitary selection of grapevine, commercially available antibodies were evaluated and conditions ...for routine ELISA testing were optimized. Extraction procedure with Granex 91 - special machine, which is used in routine testing in Slovenia, was compared with grinding samples in mortar. Three different extraction buffers were applied in order to overcome the inconvenience of using more than one extraction procedure when testing grapevine material for several viruses in a routine large-scale testing scheme. Results were verified with Western blot and immuno-electron microscopy. The best results were obtained using extraction buffer with unknown composition (pH 9.0) from BIOREBA kit for GLRaV-2. Other extraction buffers gave less positive samples and they are not convenient for routine testing where extraction with Granex is done. Both viruses, GLRaV-2 and GVB were found in Slovenia, but they couldn't be correlated with rougose wood disease that appears on indigenous cultivar Refošk grafted on ‘SO4’ from collection vineyard in Komen.
Z namenom, da bi uvedli testiranje virusa Grapevine leafroll-associated virus-2 (GLRaV-2) in virusa Grapevine virus B (GVB) v zdravstveno selekcijo vinske trte, smo testirali komercialno dostopna protitelesa in optimizirali pogoje za rutinsko testiranje v ELISA. Primerjali smo ekstrakcijo s strojem Granex 91, ki ga v Sloveniji uporabljajo v rutinskem testiranju, z ekstrakcijo v terilnici. Da bi poenotili ekstrakcijo različnih virusov v obsežnem rutinskem testiranju, smo preizkusili tri različne ekstrakcijske pufre. Rezultate smo preverili z imunskim pivnikom (Western blot) in imunsko elektronsko mikroskopijo. Najboljše rezultate smo dobili pri vzorcih ekstrahiranih s pufrom nepoznane sestave iz kita za določevanje GLRaV-2 proizvajalca BIOREBA. Z ostalima pufroma smo dobili manj pozitivnih vzorcev, zato menimo, da pufra nista primerna za ekstrakcijo vzorcev v rutinskem testiranju kjer se uporablja stroj Granex 91. Ugotovili smo prisotnost obeh virusov v Sloveniji, nismo pa uspeli dokazati povezave teh dveh virusov z razbrazdanjem lesa, ki se pojavlja na trsih domače sorte Refošk cepljenih na podlago ‘SO4’ iz kolekcijskega vinograda v Komnu.
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