magistrsko delo
Povzetek
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.
Ključne besede
biotehnologija;hmelj;suspenzijske kulture;viroid CBCVd;polarnost verig;
Podatki
Jezik: |
Slovenski jezik |
Leto izida: |
2022 |
Tipologija: |
2.09 - Magistrsko delo |
Organizacija: |
UL BF - Biotehniška fakulteta |
Založnik: |
[K. Jamnik] |
UDK: |
601.4:577.21:633.791:632.38(043.2) |
COBISS: |
125261571
|
Št. ogledov: |
56 |
Št. prenosov: |
5 |
Ocena: |
0 (0 glasov) |
Metapodatki: |
|
Ostali podatki
Sekundarni jezik: |
Angleški jezik |
Sekundarni naslov: |
Initiation of cell suspension cultures for three hop cultivares (Humulus lupulus L.) to support studies on interaction with the viroid CBCVd |
Sekundarni povzetek: |
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. |
Sekundarne ključne besede: |
biotechnology;hops;suspension cultures;viroid CBCVd;strand polarity;RT-qPCR; |
Vrsta dela (COBISS): |
Magistrsko delo/naloga |
Študijski program: |
0 |
Konec prepovedi (OpenAIRE): |
1970-01-01 |
Komentar na gradivo: |
Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije |
Strani: |
1 spletni vir (1 datoteka PDF (XII, 66 f., [6] f. pril.)) |
ID: |
16400954 |