magistrsko delo
David Snoj (Author), Jana Murovec (Mentor), Katarina Rudolf Pilih (Co-mentor)

Abstract

Belo zelje (Brassica oleracea) je kulturna rastlina iz družine križnic (Brassicaceae). Je ena izmed najpomembnejših zelenjadnic v Sloveniji. Zaradi potrebe po domačih hibridnih sortah, poteka na Biotehniški fakulteti v Ljubljani žlahtnjenje zelja. Za pridobitev hibridnega semena potrebujemo dve čisti starševski liniji. Glede na teorijo o pozitivni heterozi producirajo linije, ki so si genetsko najbolj oddaljene, najuspešnejše hibride. Za določitev kombinacijskega potenciala smo opravili genotipizacijo dvaindvajsetih čistih linij zelja, pridobljenih na Biotehniški fakulteti v Ljubljani. Genotipizacija je potekala s šestnajstimi mikrosatelitnimi markerji. Najprej smo izolirali DNK čistih linij, izmerili koncentracijo in vzorce ustrezno redčili. Opravili smo testiranje SSR markerjev in poskusili optimizirati PCR reakcijo. Genotipizacijo smo izvedli s postopkom PCR in kapilarno elektroforezo. Rezultate smo ovrednotili in statistično obdelali s programom Genemapper. Sočasno smo preskusili uporabnost metod hkratnega pomnoževanja in direktni PCR z namenom nižanja stroškov genotipizacije. Mikrosatelitni markerji uporabljeni v raziskavi kažejo nizko stopnjo polimorfizma. Kljub temu so primerni za nadaljnjo uporabo. Kombinacije čistih linij pod oznakami 278 x 461, 317 x 461, 441x 459 so si genetsko najbolj oddaljene in imajo največji potencial za pridobivanje hibridov z močno izraženimi želenimi lastnostmi. Tudi kombinacije ostalih linij smo razvrstili glede na njihovo genetsko oddaljenost. Metodi hkratnega pomnoževanja in direktni PCR sta se izkazali za uspešni ob upoštevanju dejstva, da sta bili izvedeni le na majhnem številu vzorcev. Potrebno bi bilo nadaljnje testiranje in optimizacija, da bi lahko z gotovostjo trdili, da sta metodi zanesljivi.

Keywords

genotipizacija;Brassica oleracea;belo zelje;SSR;PCR;hkratno PCR pomnoževanje;kapilarna elektroforeza;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [D. Snoj]
UDC: 601.4:577.21:606:631.528:635.342(043.2)
COBISS: 9330297 Link will open in a new window
Views: 844
Downloads: 282
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Other data

Secondary language: English
Secondary title: Genotyping of selected cabbage lines (Brassica oleracea L. var. capitata L.) with microsatellite markers
Secondary abstract: White cabbage (Brassica oleracea) is a cultural plant from the family of Brassicaceae. It is one of the most important vegetables in Slovenia. The Biotechnical Faculty in Ljubljana has been breeding cabbage due to the need for hybrid varieties. To obtain hybrid seed we need to cross two homozygous parental lines. According to the theory of positive heterosis, the lines that are genetically most distinct have the biggest potential to produce a successful hybrid. In order to determine the combination potential, genotyping of twenty-two homozygous lines of cabbage was performed. The samples were collected at the Biotechnical Faculty in Ljubljana. Genotyping was carried out with sixteen microsatellite markers. First, the DNK of the homozygotes lines was isolated, the concentration of DNK was measured, and the samples diluted accordingly. We tested the SSR markers and tried to optimize the PCR reaction. The genotyping was carried out by the PCR procedure and capillary electrophoresis. The results were evaluated and statistically processed with the GeneMapper program. At the same time, we tested the usefulness of the Multiplex and Direct PCR method in order to reduce the costs of genotyping. Microsatellite markers that we used showed low levels of polymorphism. Nevertheless, they were suitable for further use. The combinations of homozygous parental lines under the codes of 278 x 461, 317 x 461, 441x 459 are genetically most distinct and have the greatest potential for obtaining successful hybrids. We also ranged combinations of parental lines by their genetic differences. The multiplex and direct PCR methods proved to be successful given the fact that they were performed on only a small number of samples. Further testing and optimization are needed so that we can firmly assert that the methods are reliable.
Secondary keywords: genotipization;white cabbage;multiplex PCR;kapillary electrophoresis;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za agronomijo
Pages: VIII, 45 f.
ID: 11258235