Identifikacija virusov pri klonu vinske trte (Vitis vinifera L.) sorte malvazija ter njihova eliminacija s termoterapijo

Flajnik, Hana (author), Štajner, Nataša (mentor), Miljanić, Vanja (comentor)

Vinska trta Vitis vinifera L. je ena ekonomsko najpomembnejših kmetijskih rastlin. Zaradi vegetativnega razmnoževanja je nagnjena k okužbam z virusi in viroidi, ki vplivajo na njeno rast in pridelek. Z metodama RT-PCR in RT-qPCR smo detektirali sedem virusov od skupno 15 virusov, ki so vključeni v obvezen in priporočen program nadzora nad infekcijskimi boleznimi. Z uporabo termoterapije in vivo in kulture vršičkov in vitro smo popolnoma eliminirali tri viruse, eliminacija ostalih virusov je bila delno uspešna. Dodatno smo detektirali virus GAMaV, ki je bilo tokrat prvič detektirano v Sloveniji. Z metodo RT-qPCR smo v času pred in po termoterapiji analizirali profile izražanja genov DCL, AGO in RDR, ki so vključeni v mehanizem RNAi ob aktivaciji z visoko temperaturo. Izražanje preučevanih genov je bilo signifikantno zmanjšano, kar predstavlja podlago za nadaljnje študije vpliva visoke temperature na osrednje proteine RNAi.

• Language: Slovenian
• Keywords: vinska trta, Vitis vinifera L., virusi, eliminacija virusov, termoterapija, RT-PCR, RT-qPCR, tkivna kultura vršičkov, RNAi, DCL, AGO, RDR
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Year: 2023
• PID: 20.500.12556/RUL-151463
• COBISS.SI-ID: 167484419
• Publication date in RUL: 07.10.2023

Secondary language

• Language: English
• Title: Identification of viruses in cv. Malvasia (Vitis vinifera L.) and their elimination by thermotherapy

Abstract

Grapevine Vitis vinifera L. is one of the most economically important crops. Due to vegetative reproduction, it is prone to infections with viruses and viroids, which affect its growth and yield. Using RT-PCR and RT-qPCR, we detected seven viruses out of 15 viruses included in the mandatory and recommended infectious disease surveillance programme. Using thermotherapy in vivo and shoot tip culture in vitro, we eliminated three viruses, the elimination of the remaining viruses was partially successful. In addition, we detected the GAMaV virus, for which this work represents the first detection in Slovenia. Using the RT-qPCR method, we analysed the expression profile of DCL, AGO and RDR genes involved in the RNAi mechanism at high temperature activation during the pre- and post-thermotherapy period. The expression of the studied genes was significantly lower, providing the basis for further studies of the effect of high temperature on the central RNAi proteins.

• Keywords: grapevine, Vitis vinifera L., viruses, virus elimination, thermotherapy, RT-PCR, RT-qPCR, shoot tip culture, RNAi, DCL, AGO, RDR