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Ugotavljanje odpornosti, tolerance in občutljivosti različnih kultivarjev hmelja (Humulus lupulus) na viroide HLVd, HSVd in CBCVd
ID Lap, Patricija (Author), ID Jakše, Jernej (Mentor) More about this mentor... This link opens in a new window, ID Volk, Helena (Co-mentor)

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Abstract
Slovenske nasade hmelja ogrožajo trije različni viroidi: hmeljni latentni viroid (HLVd, angl. hop latent viroid), viroid zakrnelosti hmelja (HSVd, angl. hop stunt viroid) in viroid razpokanosti skorje agrumov (CBCVd, angl. citrus bark cracking viroid). Namen te magistrske naloge je bil pregledati večje število kultivarjev (v nadaljevanju bomo uporabljali besedo sorta) hmelja na občutljivost za okužbo z viroidi (HLVd, HSVd in CBCVd), ki ogrožajo slovenska hmeljišča. Tekom poskusa smo gojili različne sorte hmelja v in vitro razmerah: pripravili smo gojišče, subkultivirali rastline na sveža gojišča, spremljali rast rastlin. Pripravili smo transkripte viroidov, izvedli validacijo uspešnosti postopka in ustreznosti transkriptov z napravo Agilent Bioanalyser 2100 in nato rastline hmelja v tkivnih kulturah inokulirali z viroidnimi transkripti in dokumentirali rast ter razvoj inokuliranih in neinokuliranih rastlin. Po osmih tednih je sledilo potrjevanje uspešnosti inokulacije hmelja z metodo RT-PCR in gelsko elektroforezo. Naši podatki so pokazali, da smo z našo novo uveljavljeno metodo inokulacije viroidov uspešno okužili vse sorte hmelja z uporabo transkriptov viroidov. Zanimivo je, da je bila stopnja okužbe najvišja pri transkriptih HSVd (100 % za vseh 13 sort), sledi CBCVd (100 % za 9 sort in 75 % za 4 sorte) in najnižja za HLVd (75 % za 3 sorte, 50 % za 5 sort in 25 % za 3 sorte). Doslej nismo opazili razlik v rasti in razvoju inokuliranih in neinokuliranih rastlin in pri testiranih sortah nismo odkrili morebitne odpornosti ali tolerance. Rastline sorte 'Wye Target' in 'Bobek' so bile okužene s HLVd že pred začetkom našega dela in tako v tem primeru ni šlo za rastline brez virusov in viroidov.

Language:Slovenian
Keywords:hmelj, tkivne kulture, viroidi CBCVd, HLVd, HSVd, inokulacija, RT-PCR
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2023
PID:20.500.12556/RUL-151823 This link opens in a new window
COBISS.SI-ID:169499651 This link opens in a new window
Publication date in RUL:21.10.2023
Views:418
Downloads:66
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Secondary language

Language:English
Title:Determining the resistance, tolerance and sensitivity of different hop cultivars (Humulus lupulus) to viroids HLVd, HSVd and CBCVd
Abstract:
Slovenian hop plantations are threatened by three different viroids: hop latent viroid (HLVd), hop stunt viroid (HSVd) and citrus bark cracking viroid (CBCVd). The purpose of this master's thesis was to examine a large number of hop cultivars for susceptibility to infection with viroids (HLVd, HSVd and CBCVd), which threaten Slovenian hop farms. During the experiment, we grew different hop cultivars in in vitro conditions: we prepared the culture medium, subcultivated the plants onto fresh culture media, and monitored the growth of the plants. We prepared viroid transcripts, validated the performance and relevance of the transcripts with an Agilent Bioanalyser 2100 device, and then infected hop plants in tissue cultures with viroid transcripts and documented the growth and development of infected and uninfected plants. After eight weeks, confirmation of the success of hop infection followed with the RT-PCR method and gel electrophoresis were conducted. Our data showed that using our newly established method of viroid inoculation we have successfully infected all hop cultivars using viroid transcripts. Interestingly, the rate of infection was the highest for HSVd transcripts (100 % for all 13 cultivars), followed by CBCVd (100 % for 9 cultivars and 75 % for 4 cultivars) and the lowest for HLVd (75 % for 3 cultivars, 50 % for 5 cultivars and 25 % for 3 cultivars). Up to date, we did not observe any difference in growth and development of infected and uninfected plants and we did not discover any possible resistance or tolerance in tested varieties. Plants of cultivars 'Wye Target' and 'Bobek' were already infected with HLVd before the start of our work and in this case the plants were not virus and viroid free.

Keywords:hops, tissue culture, viroids CBCVd, HLVd, HSVd, inoculation, RT-PCR

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