magistrsko delo
Katja Jamnik (Author), Jernej Jakše (Mentor), Helena Volk (Co-mentor)

Abstract

Pridelavo hmelja (Humulus lupulus L.) ogrožajo številne bolezni, med drugimi tudi viroidi. Viroid CBCVd na hmelju povzroča hudo viroidno zakrnelost hmelja. Za študije interakcij med viroidom CBCVd in celicami hmelja smo poskušali vzpostaviti suspenzijske kulture treh kultivarjev hmelja, Celeia, Styrian Cardinal in Styrian Wolf. Pri kultivarju Styrian Cardinal vzpostavitev suspenzijskih kultur kalusa ni bila uspešna. Uspešno smo vzpostavili suspenzijske kulture kultivarjev Celeia in Styrian Wolf. Pripravili smo dimerne in vitro transkripte viroida CBCVd in razvili metodo za vzpostavitev suspenzijskih kultur, okuženih z viroidom CBCVd. Spremljali smo rast v suspenzijskih kulturah, akumulacijo sveže biomase ter živost celic. Z meritvami nismo uspeli potrditi negativnega vpliva prisotnosti viroida CBCVd na živost celic in rast v suspenzijskih kulturah. Z metodo RT-qPCR smo preverjali uspešnost okuževanja suspenzijskih kultur. Rezultati RT-qPCR kažejo, da smo uspešno okužili celice v suspenzijskih kulturah občutljivega kultivarja Celeia in tudi kultivarja Styrian Wolf, ki sodi med domnevno odporne kultivarje. Preverjali smo, kdaj se v suspenzijskih celicah kultivarja Celeia prične sinteza negativne verige viroida, ki potrjuje, da se viroid začne aktivno podvojevati. Iz rezultatov lahko sklepamo, da se podvojevanje viroida CBCVd začne okoli dva tedna po vzpostavitvi okuženih suspenzijskih kultur. Z vzpostavitvijo suspenzijskih kultur, okuženih z viroidom CBCVd, smo pridobili model za preučevanje interakcij med celicami hmelja in viroidom CBCVd.

Keywords

biotehnologija;hmelj;suspenzijske kulture;viroid CBCVd;polarnost verig;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [K. Jamnik]
UDC: 601.4:577.21:633.791:632.38(043.2)
COBISS: 125261571 Link will open in a new window
Views: 56
Downloads: 5
Average score: 0 (0 votes)
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Other data

Secondary language: English
Secondary title: Initiation of cell suspension cultures for three hop cultivares (Humulus lupulus L.) to support studies on interaction with the viroid CBCVd
Secondary abstract: Hop (Humulus lupulus L.) production is threatened by many pathogens, including viroids. Viroid CBCVd causes severe hop stunt disease. For studies of the interactions between the CBCVd viroid and hop cells, we attempted to establish suspension cultures of three hop cultivars, Celeia, Styrian Cardinal, and Styrian Wolf. The initiation of suspension cultures of the Styrian Cardinal cultivar was unsuccessful. We have successfully established suspension cultures of cultivars Celeia and Styrian Wolf. Dimeric in vitro transcripts of viroid CBCVd were used for establishing infected suspension cultures. We sought to determine the effects of CBCVd infection on cell viability and biomass accumulation. No differences in cell viability and biomass accumulation were observed between viroid-free and CBCVd-infected cell suspension cultures. RT-qPCR was used to confirm the presence of CBCVd RNA in samples from suspension cultures. The RT-qPCR results indicate that we successfully infected cells in suspension cultures of the susceptible cultivar Celeia and the cultivar Styrian Wolf, which is among the supposedly resistant cultivars. By regularly sampling biomass from infected suspension cultures of Celeia cultivar, we determined the occurrence of viroid RNA of negative polarity. Results indicate that synthesis of the negative strands of the viroid CBCVd begins in the cells of the suspension cultures at approximately two weeks postinfection. By establishing suspension cultures infected with the viroid CBCVd, we obtained a model to study the interactions between hop cells and the CBCVd viroid.
Secondary keywords: biotechnology;hops;suspension cultures;viroid CBCVd;strand polarity;RT-qPCR;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Pages: 1 spletni vir (1 datoteka PDF (XII, 66 f., [6] f. pril.))
ID: 16400954