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The indigenous vineyard mycobiota contribute both to wine quality and vineyard sanitary status. Wines made from same grape variety but from different geographical locations are appreciated for their diversity. Because no information on indigenous mycobiota of Croatian grapevines is available, the aim of the present study was to start filling this knowledge gap by characterizing the indigenous mycobiota of Maraˇstina variety. The use of metataxonomic approach has enabled the identification of 25 different fungal genera present on Maraˇstina grape berries collected from 11 vineyards located within the Croatian coastal winegrowing region of Dalmatia (Northern Dalmatia, Dalmatian hinterland, Central and Southern Dalmatia). The substantial regional and local scale differences in their distribution were observed. Overall, Aureobasidium was the dominant genus followed by Cladosporium and Metschnikowia. Botrytis and Plenodomus were associated with the vineyards located in Central and Southern Dalmatia, whereas Pichia was associated with Northern Dalmatia vineyards. The largest abundance of Buckleyzyma, Cladosporium, Eremothecium, Fusarium, Papiliotrema, and Rhodotorula was observed in Dalmatian hinterland. Moreover, data suggested that climate conditions and soil type partially influenced the distribution of fungal communities. The local-scale differences emerged also for the physicochemical characteristics of fresh musts. The high malic acid content supported the development of Metschnikowia, and inhibited Fusarium growth, whereas a positive correlation between Erysiphe and pH values was observed. Sporobolomyces and Cystobasidium were negatively associated with high glucose concentration. The revealing of Maraˇstina indigenous mycobiota provided information on the members of fungal community negatively influencing the grapevine sanitary status as well as those which could be employed in disease biocontrol.

Wine is the result of alcoholic fermentation carried out principally by Saccharomyces cerevisiae strains which transform sugars present in grape must into ethanol, CO2 and other compounds. Differently from S. cerevisiae which is scarcely present on harvested grapes, non- Saccharomyces yeasts are highly abundant which gives them the opportunity to begin fermentation. Since use of commercial yeasts may negatively affect the aromatic complexity of wine, new trends in winemaking have emphasised the concept of microbiological terroir thus stimulating the use of indigenous starters from different regions, including non-Saccharomyces yeasts aimed to increase organoleptic complexity of wines. This study focused on the oenological characterization of non- Saccharomyces yeasts isolated from 11 Maraština vineyards located within the Croatian coastal winegrowing region of Dalmatia aimed to discover new candidate strains for development as starter cultures. Among the 257 isolates, 27 different species from 16 genera were identified by sequencing the ITS1-5.8S-ITS2 region. The species from Aureobasidium genus (Aureobasidium pullulans, Aureobasidium melanogenum, and Aureobasidium namibiae) largely prevailed, followed by Metschnikowia chrysoperlae, Hanseniaspora uvarum and Lachancea thermotolerans strains. All isolates were evaluated for their ethanol and SO2 resistance, ability to grow on different temperatures, H2S and acetic acid production, as well as hydroxycinnamic acid decarboxylase (HCDC) activity. Furthermore, the 75 best performing strains were screened for the activity of 19 enzymes using API-ZYM test system with the emphasis on the enzymes related to wine aroma such as esterase, esterase lipase, lipase, leucine acrylamidase, valine acrylamidase, cystine acrylamidase, and β-glucosidase. Various strains from Lachancea, Hanseniaspora, Metschnikowia, Pichia, Hyphopichia and Starmerella genera showed high SO2 (up to 250 mg/L) and ethanol (up to 12%) tolerance, low acetic acid and medium-low H2S production. The HCDC activity was detected in several Metschnikowia and Aureobasidium isolates. Most of the isolates demonstrated high leucine and valine acrylamidase activity whereas several Hanseniaspora uvarum, Lachancea thermotolerans and Metschnikowia chrysoperlae strains were characterized by high β- glucosidase activity thus allowing the selection of the most promising strains for further development as starter cultures to produce aromatically distinct wines.

Spontaneous fermentation in winemaking is a complex microbial reaction that involves influences from many factors, including the microbiota, the grape variety, climatic and soil conditions, and vinification technology. The perceived aroma of the produced wine is the result of interactions between all the volatile and nonvolatile metabolites. The study aimed to investigate the volatile metabolite profile of „Maraština“ wines produced by spontaneous fermentation, thus supporting the concept of terroir impact on the volatile aromas of the wine. The „Maraština“ grapes were harvested in 10 vineyards from two Croatian vine-growing regions: Northern Dalmatia and Central and Southern Dalmatia. A total of 10 experimental fermentation trails in three repetitions were set up on the laboratory scale. Volatile compounds from the wine samples were isolated by solid-phase extraction (SPE) and analyzed by an untargeted approach using two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GCxGC-TOF-MS) and a targeted approach by gas chromatography-tandem mass spectrometry (GC- MS/MS). Comprehensive two-dimensional GCxGC analysis provided a detailed characterization of total volatile metabolites in the wines due to its excellent separation ability. More than 900 compounds were detected after untargeted profiling and 222 of them were identified or tentatively identified by comparison of experimental retention indices and mass spectra with literature data from the NIST database. A total of 56 volatile compounds were identified and quantified using GC-MS/MS analysis, including terpenes (14), C13- norisoprenoids (3), esters (18), alcohols (4), phenols (4), aldehydes (2), ketones (2), acids (4), lactones (4) and indole (1). The Fischer ratio was useful for establishing the compounds responsible for the main differences between grapes harvested from different subregions. „Maraština“ wines from the Northern Dalmatia subregion had a significantly higher concentration of total volatile compounds than the wines from the Central and Southern Dalmatia subregion, especially the following compounds: cis-Rose oxide, trans-Rose oxide, β-Damascenone, TDN, ethyl leucate, diethyl succinate, phenylacetaldehyde, benzaldehyde and octanoic acid.

Dehydration or drying of grapes is one of the most important steps in the production of Croatian traditional dessert wine Pro&scaron, ek. The natural sun drying of grapes is the traditionally used method in Pro&scaron, ek production. Alternative methods, such as dehydration under controlled conditions, have been studied as safer and faster methods than the traditional sun drying but without precise knowledge of the effect on volatile compounds. The objective of this work was to study how dehydration of grapes carried out in a greenhouse and an environmentally controlled chamber impacts on the free and glycosidically bound volatile compounds of native grape cv. &lsquo, Mara&scaron, tina&rsquo, The 36 volatile compounds were identified and quantified using headspace solid-phase micro extraction coupled with gas chromatography-mass spectrophotometry (HS-SPME-GC/MS). The results showed that the aroma profile of dehydrated grapes was significantly different from that of fresh grapes. Regarding free forms, significant increases in the concentration of 2-methyl-1-propanol, 1-butanol, 2-hexen-1-ol, 1-hexanol, ethyl hexanoate, hexyl acetate, o-cymene, linalool oxide, and terpinen-4-ol and geraniol were found in greenhouse-dried grapes, whereas increases in cis-limonene-epoxide, trans-limonene epoxide, and &gamma, hexalactone were higher in chamber-dried grapes compared to greenhouse-dried grapes. Glycosidically bound forms of o-cymene, linalool oxide, linalool, and terpinen-4-ol were increased in both types of drying, whereas &beta, damascenone was increased only in greenhouse-dried grapes.

Vinogradarstvo i vinarstvo na području Hrvatske ima dugu tradiciju uzgoja. U prošlosti je postojao velik broj autohtonih sorata, međutim pojavom bolesti koje su opustošile vinograde, došlo je do gubljenja pojedinih sorata. Veliki dio sorata danas je ipak očuvan, a zadnjih godina dolazi do njihove revitalizacije i povećanja proizvodnje sadnog materijala autohtonih sorata. Veliki problem danas predstavlja nekvalitetan sadni materijal zaražen virusima i ostalim biljnim patogenima. Razlog tomu je najniža kvaliteta sadnog materijala autohtonih sorata, bez provedene zdravstvene selekcije. Ranijim istraživanjima potvrđeno je da prisutnost virusa utječe na gospodarske karakteristike: smanjenje prinosa, smanjenje sadržaja šećera, slabiju obojenost i veću količinu kiselina u odnosu na zdrave trsove. Postoji više virusa, a naše istraživanje se bavi uvijenosti lista vinove loze pridruženom virusu 3 (Grapevine leafroll-associated virus 3, GLRaV-3), koji je najviše zastupljen na području Dalmacije. Pokusni nasad Baštica je nastao nakon pozitivne masovne selekcije sorata Plavac mali, Plavina, Pošip, Grk, Maraština, Vugava i Debit. U nasadu su posađeni vegetativni potomci trsova koji nisu bili zaraženi s virusom. U međuvremenu je na određenom broju trsova došlo je do zaraze, što je potvrđeno nedavnim ponovnim testiranjem pomoću serološke metode (Elisa test). Ovim istraživanjem obuhvaćeni su zaraženi i zdravi trsovi kako bi se pratilo postoji li među njima razlika u morfološkim i gospodarskim svojstvima tijekom vegetacije. Istraživane sorte bile su: Plavina, Plavac mali, Maraština, Pošip i Vugava. U istraživanju se vizualno promatranje i označavanje vršilo po BBCH skali tijekom cijele vegetacije, mjerena kvaliteta prinosa (sadržaj šećera u moštu, količina ukupnih kiselina), uvometrija i mehanička sastav grozda. Temeljem dobivenih rezultata može se zaključiti kako jedino kod zaraženih trsova sorte Maraština uočavamo negativne posljedice zaraženosti trsova s virusima u morfološkim i gospodarskim značajnim svojstvima, dok kod ostalih sorata u istraživanju nisu pronađene značajnije razlike. In Croatia, viticulture and viniculture have a long tradition. In the past, there was a large number of autochthonous varieties, but the occurrence of diseases devastating the vineyards resulted in the loss of some varieties. A great deal of the varieties is still preserved today, and in recent years their revitalization and an increase of the production of autochthonous seedlings is under way. A big problem today is the lower quality planting material infected with viruses and other plant pathogens. The reason for this is the lowest quality of autochthonous seedlings, without a conducted sanitary selection. Earlier studies have confirmed that the presence of viruses affects the economic characteristics: reducing yields, reducing sugar content, weaker coloration and higher acidity in comparison to healthy vines. There are several viruses, and our research deals with curvature of leafs, associated with virus 3 (Grapevine leafroll-associated virus 3, GLRaV-3), most commonly found in Dalmatia. The experimental plantation Baštica was created after a positive mass selection of the Plavac mali, Plavina, Pošip, Grk, Maraština, Vugava and Debit varieties. Vegetative seedlings of plants that were not infected with the virus were planted in the plantation. Meanwhile, a certain number of plants have had an infection, as confirmed by a recent retest using a serological method (ELISA test). This research included infected and healthy vines to determine whether there is a difference in morphological and economic characteristics between them during vegetation. The investigated varieties were: Plavina, Plavac mali, Maraština, Pošip and Vugava. In the study, visual inspection and marking was performed according to the BBCH scale over the whole vegetation, as well as the measuring of yield quality (sugar content in the must, the total amount of acids), the uvometry and the mechanical composition of the grape. Based on the results obtained, it can be concluded that only in the case of infected vines of the variety Maraština the negative consequences of infection of viruses are noticed in morphological and economic significant properties, while in the case of other varieties in the study no significant differences were found.

Uzgoj vinove loze na području Dalmacije ima tradiciju dužu od dva tisućljeća. Pregledom povijesnih zapisa o uzgoju istraživanih sorata na području Dalmacije većina sorata se po prvi put spominje u 19.st. Iznimku čini sorta Tribidrag o čijem uzgoju nalazimo prvi spomen u 16.st. Većina istraživanih sorata se u značajnoj mjeri uzgajala i u predfiloksernom razdoblju te su i danas gospodarski iznimno značajne na području Dalmacije, ali i Republike Hrvatske u cjelini.

Uzgoj vinove loze na području Dalmacije ima tradiciju dužu od dva tisućljeća. Pregledom povijesnih zapisa o uzgoju istraživanih sorata na području Dalmacije većina sorata se po prvi put spominje u 19.st. Iznimku čini sorta Tribidrag o čijem uzgoju prvi spomen nalazimo u 16.st. Većina istraživanih sorata se u značajnoj mjeri uzgajala i u predfiloksernom razdoblju te su i danas gospodarski iznimno značajne na području Dalmacije, ali i Republike Hrvatske u cjelini., Tradition of grape wine cultivation in Dalmatia lasts over two millenniums. Overviewing the historical accounts on the production of researched varieties in Dalmatia, most of the varieties are mentioned in the 19th century. Variety Tribidrag is an exception as accounts on its cultivation date to 16th century. Most of the researched varieties have been significantly cultivated during the pre-phylloxera period and are still exceptionally economically important in Dalmatia and Republic of Croatia as a whole.

Pošip i Maraština su bijele autohtone sorte vinove loze koje se uzgajaju na području Dalmacije. Klonska selekcija ovih sorata započeta 2006. godine. Odabrani su trsovi sa pozitivnim karakteristikama u proizvodnim nasadima ovih sorata te oni koji su pokazali odsustvo gospodarski štetnih virusa. Njihovo vegetativno potomstvo posađeno je u pokusni nasad „Baštica“ 2007. godine. Sorta Pošip zastupljena je sa 19, a Maraština sa 7 klonskih kandidata, a svaki klonski kandidat zastupljen je sa minimalno 10 trsova koji su ušli u rod 2009. godine. U ovome radu cilj istraživanja bio je odrediti razlike u najvažnijim ampelografskim karakteristikama između klonskih kandidata sorata Pošip i Maraština u 2010. godini. Na njima je provedena standardna ampelografska evaluacija koja obuhvaća vizualnu evaluaciju, utvrđivanje najvažnijih gospodarskih karakteristika: prinosa po trsu, elemenata rodnosti, sadržaja šećera i kiselina u moštu, pH vrijednosti mošta, mehaničkog sastava grozda i bobica, te uvometrijska istraživanja. Provedena je i odgovarajuća obrada podataka (analiza varijance) s ciljem utvrđivanja razlika između klonskih kandidata za sva mjerena svojstva. Rezultati pokazuju kako između klonskih kandidata sorte Pošip postoji značajna razlika u svim promatranim gospodarski važnim karakteristikama zbog dok su kod Maraštine uočene razlike znatno manje. Za izdvajanje klonova nužno je nastaviti istraživanje još dvije godine.

In Croatia, like in all mediterranean countries, there is a tradition centuries old in grape and wine production. A whole set of geografical and climate characteristics, next to historical circumstances, caused a diversity in grapevine’s cultivars in such a small territory. Clone selection of Dalmatian autochtohonous cultivars started in the year 2006 because of many positive, economically important characteristics. Ten candidate clones of each mother vines were chosen and planted in the experimental vineyard in Baštica in the year 2006. Candidate clones were tested by the ELISA test for the presence of economically important, legally requested viruses. In this diploma thesis the aim was to determine variability of the amphelographic, important characteristics between clone candidates of cultivars Pošip and Maraština. In 2010 a detailed amphelographic evaluation of candidate clones was carried out. Amphelographic evaluation includes a visual evaluation, identifying the most important economically characteristics : yield per vine, elements of fertility at grape varietas, sugar content and acidity of wine ; pH value and mechanical composition of grape and berries. An appropriate data processing (analysis of variance ) was carried out to determine the differences between clone candidates. Reduced variability between candidate clones of Maraština could be attributed to the fact that this candidates were selected from only one location.The results of the analysis of variance showed that there was a great variability (a statistically, highly significant differences) between candidates of cultivar Pošip, in all the economically important characteristics. This confirms that a large number of candidate clones were selected from several locations.

Polifenolni sastav grožđa i vina predmet su brojnih istraživanja zahvaljujući njihovom utjecaju na organoleptična svojsva vina ali i pozitivnom učinku kojeg imaju na ljudsko zdravlje. U grožđu i vinu dijele se na neflavonoide (hidroksibenzojeve kiseline, hidroksicimetne kiseline i stilbeni) i flavonoide (flavonoli, flavan-3-oli, antocijanini i njihovi polimeri). Zavisno od pojedinačnih koncentracija te međusobnog odnosa utječu na boju, gorčinu, trpkoću i potencijal starenja vina. Dozrijevanje grožđa podrazumijeva široku paletu fizioloških i biokemijskih procesa koji se intenziviraju od trenutka šare grožđa, a završavaju sa punom zrelosti bobice, te su u konačnici i odgovorni za postignutu kakvoću grožđa. Promjene koje nastupaju tijekom perioda dozrijevanja ne odvijaju se istovremeno, niti jednakim intenzitetom. Svaki kemijski spoj pa tako i indol-3-octena kiselina te polifenoli ima vlastitu „krivulju dozrijevanja“ na koju dodatno mogu utjecati različiti čimbenici kao što su genetske karakteristike kultivara, klimatske prilike, te agrotehnički i ampelotehnički zahvati koji se provode u vinogradu, enološki postupci tijekom vinifikacije i starenja vina, te rok berbe.

The alcoholic fermentation and maturation of white wines in Croatia is often carried out in the barrique barrels made from the Croatian oak. Scientific research in their effect on the quality of obtained white wines has been scant. Hence the goal of the present research was to determine the influence of the Croatian barrique barrels on the quality of the autochthonous cultivars of Maraština White. Barrique barrels were used made from Quercus robur and Quercus petrea, with their staves light and medium toasted. The completion of alcoholic fermentation was followed by an analysis of the basic chemical composition of the wines and the aromatic oak compounds by using the gas chromatography-mass spectrometry (GC-MS) and the high performance liquid chromatography (HPLC). The wines were sensory evaluated by the 100- point O.I.V. method. The obtained results showed concentration variations in individual aromatic compounds, linked to the type of oak and the intensity of stave toasting. Maraština wines, obtained by fermentation in the Quercus petrea barrels contained a little higher translactone concentrations compared with the wines from the Quercus robur barrels. The stave toasting intensity had the strongest impact on the increase in the oak lactone concentrations, whereas the levels of eugenol and guaiacol were rising at a slower rate. All wines contained relatively low concentrations of syringaldehyde and vanillin. The results of the sensory evaluation of Maraština wines show a better quality of samples obtained through fermentation in the Quercus petrea barrels with the medium toasted staves

Among 48 cultivars holding the „ Malvasia“ name listed in the Vitis International Variety Catalogue (http://www.genres.de/idb/vitis/vitis.htm) there are also two cultivated in Croatia – „ Malvazija istarska“ and „ Malvasia dubrovačka“ . Since recently it is known that Malvasia dubrovacka is not a unique local cultivar (genotype) but the member of the synonym group which is scattered across the Mediterranean and even at the Atlantic islands (Malvasia delle Lipari, Malvasia di Sardegna, Malvasia de Sitges, Malvasia de Tenerife, ...) (Crespan et al., 2006). So far, Malvazija istarska seems to be a unique genotype spread only in the area of peninsula Istria and north-east of Italy. According to Boursiquot et al. (2007), the real number of Malvasia cultivars is not so large as it used to be believed, i.e. they found that several so called Malvasia cultivars are synonyms of already well known cultivars. Taking into account the historical fact that „ Malvasia wine“ used to be exported from Greece to Venice for centuries and an assumption that the name „ Malvasia“ has Greek origin (according to medieval city Monemvasia), as well as the fact that by vegetative propagation cultivars can be easily disseminated and preserved over the space and time we have hypothesized that Croatian Malvasias might still be grown (under different names) in Greece or some other places along the historical naval routs. We have used an opportunity to participate in the sailing expedition „ Malvasia Mediterranea 2005“ (http://www.malvasia-mediterranea.com/) to explore and sample the tissue for DNA of the cultivars along the eastern coast of Adriatic and Ionian Sea, and south of Peloponnesus. Twenty-eight grapevine samples from Greece (islands and hinterland), more than 30 from Croatia, and several from Italian coastal regions that used to be on the ancient naval routs from Monemvasia to Venice, have been assessed by genomic DNA SSR markers and analyzed in term of genetic similarity. Similarly as in several previous studies we have found a weak relationship of actual Greek cultivars with Croatian and Italian ones. We have found only one original genotype that is nowadays present in Greece, Italy and Croatia under three different names (synonyms), Pavlos, Malvasia lunga (M. del Chianti) and Maraština, respectively.

Zaštita kontroliranog podrijetla kvalitetnih vina Debit, Maraština, Plavina i Lasina

Cilj rada je bio utvrditi unutarsortnu genetsku varijabilnost autohtonih sorata vinove loze Plavac mali, Maraština i Škrlet pomoću raznih metoda molekularnih markera (SSR, ISSR i AFLP), a kod Škrleta osim molekularnih primjenjene su ampelografske i ampelometrijske metode. Kao referentne sorte poslužili su komercijalno priznati klonovi Chardonnaya i Traminca s kojima je provedena usporedba. Varijabilnost detektirana ampelografskim metodama uspoređena je sa varijabilnošću utvrđenom molekularnim metodama. Podaci o morfološkim svojstvima analizirani su po O.I.V. sustavu deskriptora, a na temelju kvantitativnih podataka dodatno ja izračunata Euklidska distanca kao mjera fenotipske različitosti. SSR markeri su korišteni u svrhu verifikacije sorte. Molekularni podaci dobiveni AFLP analizom korišteni su za izračunavanje SM koeficijenta kao mjere genetske sličnosti. Za sorte Chardonnay, Traminac i Škrlet utvrđen je stupanj korelacije fenotipskih razlika i razlika koje proizlaze iz molekularnih podataka. Analizom molekularne varijance (AMOVA) su određene komponente varijance između grupa izdvojenih ''cluster'' analizom. Razina polimorfizma morfoloških svojstava s jedne i molekularnih markera s druge strane ukazuje na dovoljno široku bazu koja bi omogućila pronalaženje genetski divergentnih klonova, međutim njihova korelacija nije značajna. Najveću snagu detekcije unutarsortne molekularne varijabilnosti pokazali su AFLP markeri. ISSR markeri su pokazali manji stupanj detekcije unutarsortne genetske varijabilnosti u odnosu na AFLP markere. Prosječna vrijednost genetske sličnosti kod Plavca malog je relativno visoka u odnosu na ostale analizirane sorte, što ukazuje na manju unutarsortnu varijabilnost ove sorte unatoč njenom relativno velikom arealu rasprostranjenosti.

Istraživanja predstavljaju ampelografsku studiju kultivara Maraština, Bogdanuša, Vugava i Pošip (Vitis vinifera L.), a glavni je cilj detaljna ampelografska obrada i ocjena mogućnosti njihova uzgoja i širenja izvan rajonizacijom predviđenih područja. To su stare, lokalne, najvjerojatnije autohtone sorte Dalmacije. Prema botaničkim svojstvima Maraština pripada ekološko-geografskoj grupi Convarietas Orinetalis. Utvrđen su i agrobiološka, te gospodarsko - tehnološka svojstva u uvjetima Sjeverne Dalmacije. Dobiveni su povoljni rezultati, posebice glede priroda i kakvoće unatoč nepovoljnim klimatskim prilikama, pa smatramo da su svi kultivari interesantni za daljnji istraživački rad.