Analiza izražanja mikro RNA in njihovih tarč v obrambnem odzivu hmelja (Humulus lupulus L.) ob okužbi z glivo Verticillium nonalfalfae

Kolenc, Živa (author), Štajner, Nataša (mentor), Kunej, Urban (comentor)

V okviru raziskav za izvedbo magistrskega dela smo analizirali izražanje nekaterih mikro RNA (miRNA) in njihovih tarč v odpornem in občutljivem kultivarju hmelja po okužbi z glivo Verticillium nonalfalfae v štirih različnih časovnih točkah ter eksperimentalno analizirali razrez tarčnih mRNA. Iz koreninskih in stebelnih vzorcev hmelja smo izolirali celotno RNA ter izvedli metodi RT-qPCR in 5' RLM-RACE. Ugotovili smo, da se izražanje med kontrolo in okuženo rastlino kot odziv na okužbo razlikuje v vsaj eni preverjeni časovni točki pri petih od sedmih analiziranih tarč in pri treh od petih analiziranih miRNA. Pri petih tarčah in pri eni miRNA so kot odziv na okužbo prisotne tudi razlike v izražanju med kultivarjema. Potrdili smo interakcijo mRNA dejavnika odziva na avksin z miR160. S pridobljenimi rezultati smo pridobili boljše razumevanje imunskega odziva hmelja in hmeljnih mehanizmov odpornosti, ki lahko v prihodnje pomagajo pri obvladovanju bolezni verticilijske uvelosti.

• Language: Slovenian
• Keywords: hmelj, Humulus lupulus, miRNA, mRNA, Verticillium nonalfalfae, rastlinska odpornost
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Year: 2022
• PID: 20.500.12556/RUL-141615
• COBISS.SI-ID: 126122499
• Publication date in RUL: 03.10.2022

Secondary language

• Language: English
• Title: MICRO RNA AND THEIR TARGETS EXPRESSION ANALYSIS IN DEFENSE RESPONSE IN HOPS (Humulus lupulus L.) AFTER INFECTION WITH FUNGUS Verticillium nonalfalfae

Abstract

In the research for this master thesis we have analysed the expression of microRNAs (miRNAs) and their targets in the resistant and susceptible cultivars of hops after infection with fungus Verticillium nonalfalfae in four different time points and experimentally analysed the cleavage of target mRNAs. We have isolated total RNA from root and stem samples of hops and conducted RT-qPCR and 5’RLM-RACE methods. We have determined that expression between control and inoculated plant is changed in at least one tested time point in five out of seven analysed mRNAs and in three out of five analysed miRNAs. Changes in expression between cultivars due to inoculation is present in five targets and one miRNA. We have confirmed the interaction of the auxin response factor and miR160. With the gathered results we have gained better understanding of immune response in hops and hop immunity mechanisms, which can help with verticillium wilt disease damage control in the future.

• Keywords: hops, Humulus lupulus, miRNA, mRNA, Verticillium nonalfalfae, plant resistance