Molecular markers used for determination of red raspberry cultivars and tracing hypothetical medieval genotypes

Naša raziskava je imela dva dela. V 1. delu smo proučevali genetske odnose med malinjaki (Rubus idaeus L.), zbranimi okoli nekaterih karakterističnih srednjeveških gradov, kartuzijanskih samostanov in v bližnjih vaseh. V 2.delu smo enako raziskavo opravili s sedmimi novejšimi kultiviranimi sortami, ki izvirajo iz Bolgarije, Francije, Slovenije in Švice, in z metodo s prstnimi odtisi s pomočjo mikrosatelitskih markerjev identificirali sorte. Predvidevali smo, da bi genotipe hipotetičnih srednjeveških malinjakov lahko zasledili ob ostankih srednjeveških naselij. Ti genotipi so preživeli v naravnih razmerah brez generativnega razmnoževanja vsaj 3 stoletja. Analiza je temeljila na mikrosatelitskih markerjih. Identificirali smo 155 alelov, povezanih z 18 mikrosatelitskimi lokusi. Dendrogram je razvrstil analizirane genotipe iz 1. dela raziskave v 7 glavnih skupin. Analize so pokazale, da sta bila najverjetnejša srednjeveška genotipa zbrana blizu 2 kartuzijanskih samostanov: Žiče in Jurklošter. Morfološko sta si bila podobna, primitivne in bujne rasti, vendar ne povsem divja genotipa. Iz podatkov, ki bazirajo na velikosti alelov, smo izbrali specifične mikrosatelitske lokuse (6 ali 7 izbranih lokusov), za vsako od 7-ih proučevanih sort. S kombinacijo uporabljenih mikrosatelitskih lokusov, smo dobili sortno specifičen »prstni odtis« za posamezne sorte, ki je omogočil njihovo identifikacijo.Our investigation was based on a molecular study of genetic relationships amongst red raspberry (Rubus idaeus L.) genotypes collected around selected medieval castles, Carthusian monasteries and nearby villages. We assumed that the hypothetical medieval raspberry genotypes could be traced to isolated medieval settlements that used to be highly prosperous during the feudal era but were later abandoned. These genotypes survived in natural conditions without seed multiplication for at least three centuries. The molecular genetic analysis was based on micro-satellite data. We detected a total of 155 alleles on 18 microsatellite loci. The clustering method grouped the analysed genotypes into seven main clusters. The analyses indicated that the most probable medieval genotypes had been collected close to two Carthusian monasteries: Zice and Jurkloster. They were morphologically similar, vigorous, and primitive but obviously not wild genotypes. In the second part of our investigation, the molecular approach based on microsatellite data was used for the identification of seven selected raspberry varieties from different countries (Bulgaria, France, Slovenia and Switzerland). From the completed data (allele sizes) obtained with 19 optimized SSR primers, we selected the most specific microsatellite loci for each of the seven cultivars. For the identification of individual cultivars, we used six or seven selected microsatellite loci in order to obtain a variety specific fingerprint for each cultivar