Morfološka i molekularna karakterizacija potencijalnih klonova sorte vinove loze Vranac

Abstract

Mоrfоloškа i mоlеkularnа kаrаktеrizаciја pоtеnciјаlnih klоnоvа sоrtе vinоvе lоzе Vrаnаc Rezime Klonska selekcija se zasniva na unutarsortnoj genetičkoj varijabilnosti. Genetske varijacije potiču uglavnom od sponatanih, prirodnih mutacija, koje se, ukoliko su dobre, mogu registrovati i putem vegetativnog umnožavanja sačuvati. Verovatnoća pojave unutarsortne varijabilnosti povećava se starenjem vinograda, kao i kod sorti koje su zastupljene na značajnim površinama u određenom području. Sorta Vranac je najzastupljenija sorta Crne Gore koja svoje granice uzgajanja ne temelji samo na prostoru sa kog potiče, već je prisutna na području Zapadnog Balkana. Kao stara sorta podložna je spontanom mutiranju. Cilj rada ove disertacije je bio potencijalna izolacija elitnih čokota sorte Vranac, čijim bi se nastavkom kroz postupak selekcije izdvojili novi klonovi. Istraživanja su obavljena na mikrolokalitetu "Aerodrom" kompanije "13. jul - Plantaže" u vinogradu starom više od četrdeset godina. U prvoj faza klonske selekcije 2014. godine definisani su klonski kandidati sa poboljšanim proizvodnim karakteristima u poređenju sa sortom populacijom. Za eksperiment je korišćen skup od oko 10 000 čokota, od kojih je vizuelnom ocenom za dalju analizu izdvojeno 104 čokota. Sublimacijom sveukupnih analiza izdvojena su na kraju 21 potencijalna klona sorte Vranac. Tokom dvogodišnjeg perioda (2015 i 2016) proučavani su klimatski i edafski uslovi ispitivanog lokaliteta, a kod 21 selekcionisanog klona ampelografska i molekularna karakterizacija, analiza virusnih infekcija, fenološke osobine, rodnost, osobine grozda i bobice, kvalitet šire, hemijski analiza vina i degustaciona ocena vina. Analiza parametara srednjih mesečnih i godišnjih temperatura pokazala je da su temperaturni uslovi lokaliteta bili veoma povoljni za izvođenje ekperimenta. Subregion Podgorica prema vrednosti srednje vegetacione temperature (VEGT) pripada “vrućoj” klimatskoj grupi. U odnosu na vrednosti Vinklerovog indeksa (WI) nalazi se u Regionu V (C III). Količina padavina bila je takođe zadovoljavajuća. Ampelografska ispitivanja, kao i molekularna karakterizacija su pokazali da ne postoje značajne razlike između ispitivanih klonova, tako da svi oni pripadaju sorti Vranac. Razlike između klonskih kandidata bile su uočljive u privredno-tehnološkim karakteristikama grozda i bobice, kao i u hemijskim i senzornim karakteristikama vina. Ispitivani klonovi nisu pokazali razlike u odnosu na kvalitativne karakteristike mladog lastara, mladog lista, lastara, cvasti i zrelog lista, dok je neznatno odstupanje utvrđeno za osobine grozda i bobice. Molekularna analizom, na osnovu SSR markera utvrđena je sličnost između ispitivanih klonova. Na osnovu ELISA testa ustanovljeno je da su virusi GLRaV-1 i GLRaV-3 bili prisutni kod svih testiranih klonskih kandidata, dok je GFLV virus bio prisutan kod 9 potencijalnih klonova. Početak vegetacije posmatranih klonskih kantidata započinjao je od 18-20.03.2015., odnosno 16-20.03.2016. Aktiviranje pupoljaka za 2015. godinu je registrovano u periodu od 02.04-09.04., odnosno 27.03-02.04., za 2016. godinu. Početak cvetanja se tokom 2015. godine‚ odvijao u periodu od 15.05-23.05., odnosno 16.05-20.05., za 2016. godinu. Šarak grožđa je tokom 2015. godine u zavisnoti od klonskog kandidata nastupio od 13.07-22.07., odnosno od 16.07-24.07., u 2016. godini. Sazrevanje lastara je u obe ispitivane godine nastupilo gotovo identično kod svih klonova. Klonovi su prema koeficijentu potencijalne rodnosti podeljeni u tri grupe: sa niskim, srednjim i visokim, i svaka grupa brojala je po 7 klonova. U pogledu krupnoće grozda, najzastupljeniji su bili klonski kandidati sa srednje grozdovima (150-225 g), ukupno 13. Najveći prosečan prinos po čokotu i ukupan broj grozdova imao je klon V9 (24/1). Klon V10 (27/5) imao je najniže vrednosti ovih pokazatelja. Prosečna masa grozda je bila najmanja kod klona V2 (4/28), a najveća kod V6 (14/20)...

Clone selection is based on intra-varietal genetic variability. Genetic variations come mainly from spontaneous, natural mutations, which, if good, can be registered and preserved through vegetative propagation. The probability of intra-varietal variability increases with the aging of vineyards, as well as with varieties that are present in significant areas in a particular teritory. The Vranac variety is the most represented variety of Montenegro, which does not base its cultivation boundaries only on the area from which it originates, but is present in the Western Balkans. As an old variety, it is susceptible to spontaneous mutation. The aim of this dissertation was the potential isolation of elite vines of the Vranac variety, the continuation of which would single out new clones through the selection process. The research was conducted on the micro-locality "Aerodrom" of the company "July 13 - Plantations" in a vineyard more than forty years old. In the first phase of clonal selection in 2014, clonal candidates with improved production characteristics were defined in comparison with the variety population. A set of about 10,000 vines was used for the experiment, of which 104 vines were selected by visual assessment for further analysis. By sublimation of the overall analyzes, 21 potential clones of Vranac variety were isolated at the end. During the two-year period (2015 and 2016) the climatic and edaphic conditions of the examined locality were studied, and in 21 selected clones ampelographic and molecular characterization, analysis of viral infections, phenological characteristics, yielding, cluster and berry characteristics, quality of must, chemical analysis of wine and tasting evaluation of wine. The analysis of the parameters of the mean monthly and annual temperatures showed that the temperature conditions of the site were very favorable for performing the experiment. According to the value of mean vegetation temperature (VEGT), the Podgorica subregion belongs to the “hot” climate group. In relation to the values of the Winkler index (WI) it is located in Region V (C III). The amount of precipitation was also satisfactory. Ampelographic investigation, as well as molecular characterization showed that there are no significant differences between the examined clones, so they all belong to the Vranac variety. Differences between clone candidates were noticeable in the economic and technological characteristics of clusters and berries, as well as in the chemical and sensory characteristics of wine. The examined clones did not show differences in relation to the qualitative characteristics of young shoots, young leaves, shoots, inflorescences and mature leaves, while a slight deviation was found for the characteristics of clusters and berries. Molecular analysis, based on SSR markers, established similarity between the examined clones. Based on the ELISA test, it was found that the GLRaV-1 and GLRaV-3 viruses were present in all tested clone candidates, while the GFLV virus was present in 9 potential clones. The beginning of the vegetation of the observed clonal candidates started from 18- 20.03.2015., ie 16-20.03.2016. Activation of buds for 2015 was registered in the period from 02.04- 09.04., ie 27.03-02.04., for 2016. The beginning of flowering took place in 2015 in the period from 15.05-23.05., ie 16.05-20.05., for 2016. During 2015, depending on the clone candidate, a veraison of grapes appeared from 13.07-22.07., ie from 16.07-24.07., in 2016. The maturation of shoots in both examined years occurred almost identically in all clones. The clones were divided into three groups according to the potential yieding coefficient: low, medium and high, and each group had 7 clones. In terms of cluster size, the most common were clone candidates with medium clusters (150-225 g), a total of 13. Clone V9 (24/1) had the highest average yield per vine and the total number of bunches. Clone V10 (27/5) had the lowest values of these indicators. The average bunch weight was the lowest in clone V2 (4/28), and the highest in V6 (14/20)...