Comparative study of different surface sterilization treatments and optimal month for establishment of aseptic cultures of raspberry cultivars
Uporedna analiza različitih tretmana za sterilizaciju i optimalnog meseca za uspostavljanje aseptičnih kultura sorti maline
Апстракт
Removing contaminants from plant material with minimal damage to plant cells using different chemical solutions represents very important and one of the most critical steps in establishing plant tissue culture in vitro. Our study deals with the optimisation of the protocol for successful surface sterilization of initial explants in two raspberry cultivars, ‘Meeker’ and ‘Willamette’ in two experimental years. The main objective during the first experimental year was to examine the protocols for surface sterilization which included different combinations of disinfectants (mercuric chloride or commercial bleach solution each in combination with 70% ethanol). In the following year, the impact of the two most effective sterilization treatments on the establishment of aseptic cultures over the four-month period (May, June, July, and August) was investigated in both raspberry cultivars. Aseptic cultures were established on Murashige and Skoog medium (MS) containing 2.0 mg/l N6-Benzyladenine (...BA), 0.5 mg/l indole-3-butyric acid (IBA) and 0.1 mg/l gibberellic acid (GA3). The highest percentage of explants initiation in ‘Meekerʼ (79.3%) was achieved with surface sterilization in 70% ethanol (1 min) combined with 10% (v/v) commercial bleach solution (20 min), whereas sterilization in 70% ethanol (1 min) followed by HgCl2 (3 min) gave the best results (38.3%) in ‘Willametteʼ. The most optimal month for the establishment of aseptic culture was May for ‘Meeker’, while June was the most suitable month for the ‘Willametteʼ cultivar.
Eliminacija zaraze sa biljnog materijala uz minimalno oštećenje biljnih ćelija primenom različitih hemijskih rastvora je izuzetno važan i jedan od najkritičnijih koraka u postupku uspostavljanja in vitro kulture biljnog tkiva. U radu se ispituje optimizacija protokola uspešne površinske sterilizacije početnih eksplanata kod dve sorte maline, ‘Meekerʼ i ‘Willametteʼ, tokom dve eksperimentalne godine. U prvoj eksperimentalnoj godini, ispitivan je protokol za uspešnu površinsku sterilizaciju, uključujući varijacije dezinficijenata poput živinog hlora i varikine, svaki u kombinaciji sa 70% etanolom. Sledeće eksperimentalne godine, ispitivan je uticaj dva najefikasnija tretmana sterilizacije na uspešnost uspostavljanja aseptične kulture tokom četvoromesečnog perioda od maja do avgusta meseca. Aseptična kultura je uspostavljena na Murashige i Skoog podlozi (MS) koja sadrži 2.0 mg/l N6-benziladenina (BA), 0,5 mg/l indol-3-buterne kiseline (IBA) i 0,1 mg/l giberelinske kiseline (GA3). Najviši ...procenat inicijacije eksplantata kod sorte ‘Meekerʼ (79,3%) postignut je pri površinskoj sterilizaciji u 70% etanolu (1 min) u kombinacijii sa 10% (v/v) varikinom (20 min), dok je sterilizacija u 70% etanolu (1 min) praćena HgCl2 (3 min) dala najbolje rezultate (38,3%) kod sorte ‘Willametteʼ. Najoptimalniji mesec za uspostavljanje aseptične kulture bio je maj za sortu ‘Meekerʼ, dok je jun najpogodniji mesec za sortu ‘Willametteʼ.
Кључне речи:
Rubus idaeus / tissue culture / disinfectants / aseptic culture establishment / kultura tkiva / dezinfekciono sredstvo / uspostavljanje aseptične kultureИзвор:
Journal of Central European Agriculture, 2024, 25, 2, 470-480Издавач:
- Agricultural University Plovdiv, Bulgaria; University of Zagreb, Faculty of Agriculture, Croatia; University of Osijek, Faculty of Agrobiotechnical Sciences, Croatia; Institute of Agriculture and Tourism, Poreč, Croatia; University of South Bohemia, Faculty of Agriculture and Technology, Czech Republic; Hungarian University of Agriculture and Life Sciences, Georgikon Campus, Hungary; Bydgoszcz University of Science and Technology, Poland; University of Agricultural Sciences and Veterinary Medicine, Romania; University of Kragujevac, Faculty of Agronomy Čačak, Serbia; Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources, Slovakia; Agricultural Institute of Slovenia, Slovenia
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200215 (Институт за воћарство, Чачак) (RS-MESTD-inst-2020-200215)
Институција/група
Institut za voćarstvoTY - JOUR AU - Andjelic, Tatjana AU - Vujović, Tatjana AU - Jevremović, Darko AU - Tomić, Jelena AU - Radivojević, Dragan PY - 2024 UR - https://refri.institut-cacak.org/handle/123456789/890 AB - Removing contaminants from plant material with minimal damage to plant cells using different chemical solutions represents very important and one of the most critical steps in establishing plant tissue culture in vitro. Our study deals with the optimisation of the protocol for successful surface sterilization of initial explants in two raspberry cultivars, ‘Meeker’ and ‘Willamette’ in two experimental years. The main objective during the first experimental year was to examine the protocols for surface sterilization which included different combinations of disinfectants (mercuric chloride or commercial bleach solution each in combination with 70% ethanol). In the following year, the impact of the two most effective sterilization treatments on the establishment of aseptic cultures over the four-month period (May, June, July, and August) was investigated in both raspberry cultivars. Aseptic cultures were established on Murashige and Skoog medium (MS) containing 2.0 mg/l N6-Benzyladenine (BA), 0.5 mg/l indole-3-butyric acid (IBA) and 0.1 mg/l gibberellic acid (GA3). The highest percentage of explants initiation in ‘Meekerʼ (79.3%) was achieved with surface sterilization in 70% ethanol (1 min) combined with 10% (v/v) commercial bleach solution (20 min), whereas sterilization in 70% ethanol (1 min) followed by HgCl2 (3 min) gave the best results (38.3%) in ‘Willametteʼ. The most optimal month for the establishment of aseptic culture was May for ‘Meeker’, while June was the most suitable month for the ‘Willametteʼ cultivar. AB - Eliminacija zaraze sa biljnog materijala uz minimalno oštećenje biljnih ćelija primenom različitih hemijskih rastvora je izuzetno važan i jedan od najkritičnijih koraka u postupku uspostavljanja in vitro kulture biljnog tkiva. U radu se ispituje optimizacija protokola uspešne površinske sterilizacije početnih eksplanata kod dve sorte maline, ‘Meekerʼ i ‘Willametteʼ, tokom dve eksperimentalne godine. U prvoj eksperimentalnoj godini, ispitivan je protokol za uspešnu površinsku sterilizaciju, uključujući varijacije dezinficijenata poput živinog hlora i varikine, svaki u kombinaciji sa 70% etanolom. Sledeće eksperimentalne godine, ispitivan je uticaj dva najefikasnija tretmana sterilizacije na uspešnost uspostavljanja aseptične kulture tokom četvoromesečnog perioda od maja do avgusta meseca. Aseptična kultura je uspostavljena na Murashige i Skoog podlozi (MS) koja sadrži 2.0 mg/l N6-benziladenina (BA), 0,5 mg/l indol-3-buterne kiseline (IBA) i 0,1 mg/l giberelinske kiseline (GA3). Najviši procenat inicijacije eksplantata kod sorte ‘Meekerʼ (79,3%) postignut je pri površinskoj sterilizaciji u 70% etanolu (1 min) u kombinacijii sa 10% (v/v) varikinom (20 min), dok je sterilizacija u 70% etanolu (1 min) praćena HgCl2 (3 min) dala najbolje rezultate (38,3%) kod sorte ‘Willametteʼ. Najoptimalniji mesec za uspostavljanje aseptične kulture bio je maj za sortu ‘Meekerʼ, dok je jun najpogodniji mesec za sortu ‘Willametteʼ. PB - Agricultural University Plovdiv, Bulgaria; University of Zagreb, Faculty of Agriculture, Croatia; University of Osijek, Faculty of Agrobiotechnical Sciences, Croatia; Institute of Agriculture and Tourism, Poreč, Croatia; University of South Bohemia, Faculty of Agriculture and Technology, Czech Republic; Hungarian University of Agriculture and Life Sciences, Georgikon Campus, Hungary; Bydgoszcz University of Science and Technology, Poland; University of Agricultural Sciences and Veterinary Medicine, Romania; University of Kragujevac, Faculty of Agronomy Čačak, Serbia; Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources, Slovakia; Agricultural Institute of Slovenia, Slovenia T2 - Journal of Central European Agriculture T1 - Comparative study of different surface sterilization treatments and optimal month for establishment of aseptic cultures of raspberry cultivars T1 - Uporedna analiza različitih tretmana za sterilizaciju i optimalnog meseca za uspostavljanje aseptičnih kultura sorti maline EP - 480 IS - 2 SP - 470 VL - 25 DO - https://doi.org/10.5513/JCEA01/25.2.4201 ER -
@article{ author = "Andjelic, Tatjana and Vujović, Tatjana and Jevremović, Darko and Tomić, Jelena and Radivojević, Dragan", year = "2024", abstract = "Removing contaminants from plant material with minimal damage to plant cells using different chemical solutions represents very important and one of the most critical steps in establishing plant tissue culture in vitro. Our study deals with the optimisation of the protocol for successful surface sterilization of initial explants in two raspberry cultivars, ‘Meeker’ and ‘Willamette’ in two experimental years. The main objective during the first experimental year was to examine the protocols for surface sterilization which included different combinations of disinfectants (mercuric chloride or commercial bleach solution each in combination with 70% ethanol). In the following year, the impact of the two most effective sterilization treatments on the establishment of aseptic cultures over the four-month period (May, June, July, and August) was investigated in both raspberry cultivars. Aseptic cultures were established on Murashige and Skoog medium (MS) containing 2.0 mg/l N6-Benzyladenine (BA), 0.5 mg/l indole-3-butyric acid (IBA) and 0.1 mg/l gibberellic acid (GA3). The highest percentage of explants initiation in ‘Meekerʼ (79.3%) was achieved with surface sterilization in 70% ethanol (1 min) combined with 10% (v/v) commercial bleach solution (20 min), whereas sterilization in 70% ethanol (1 min) followed by HgCl2 (3 min) gave the best results (38.3%) in ‘Willametteʼ. The most optimal month for the establishment of aseptic culture was May for ‘Meeker’, while June was the most suitable month for the ‘Willametteʼ cultivar., Eliminacija zaraze sa biljnog materijala uz minimalno oštećenje biljnih ćelija primenom različitih hemijskih rastvora je izuzetno važan i jedan od najkritičnijih koraka u postupku uspostavljanja in vitro kulture biljnog tkiva. U radu se ispituje optimizacija protokola uspešne površinske sterilizacije početnih eksplanata kod dve sorte maline, ‘Meekerʼ i ‘Willametteʼ, tokom dve eksperimentalne godine. U prvoj eksperimentalnoj godini, ispitivan je protokol za uspešnu površinsku sterilizaciju, uključujući varijacije dezinficijenata poput živinog hlora i varikine, svaki u kombinaciji sa 70% etanolom. Sledeće eksperimentalne godine, ispitivan je uticaj dva najefikasnija tretmana sterilizacije na uspešnost uspostavljanja aseptične kulture tokom četvoromesečnog perioda od maja do avgusta meseca. Aseptična kultura je uspostavljena na Murashige i Skoog podlozi (MS) koja sadrži 2.0 mg/l N6-benziladenina (BA), 0,5 mg/l indol-3-buterne kiseline (IBA) i 0,1 mg/l giberelinske kiseline (GA3). Najviši procenat inicijacije eksplantata kod sorte ‘Meekerʼ (79,3%) postignut je pri površinskoj sterilizaciji u 70% etanolu (1 min) u kombinacijii sa 10% (v/v) varikinom (20 min), dok je sterilizacija u 70% etanolu (1 min) praćena HgCl2 (3 min) dala najbolje rezultate (38,3%) kod sorte ‘Willametteʼ. Najoptimalniji mesec za uspostavljanje aseptične kulture bio je maj za sortu ‘Meekerʼ, dok je jun najpogodniji mesec za sortu ‘Willametteʼ.", publisher = "Agricultural University Plovdiv, Bulgaria; University of Zagreb, Faculty of Agriculture, Croatia; University of Osijek, Faculty of Agrobiotechnical Sciences, Croatia; Institute of Agriculture and Tourism, Poreč, Croatia; University of South Bohemia, Faculty of Agriculture and Technology, Czech Republic; Hungarian University of Agriculture and Life Sciences, Georgikon Campus, Hungary; Bydgoszcz University of Science and Technology, Poland; University of Agricultural Sciences and Veterinary Medicine, Romania; University of Kragujevac, Faculty of Agronomy Čačak, Serbia; Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources, Slovakia; Agricultural Institute of Slovenia, Slovenia", journal = "Journal of Central European Agriculture", title = "Comparative study of different surface sterilization treatments and optimal month for establishment of aseptic cultures of raspberry cultivars, Uporedna analiza različitih tretmana za sterilizaciju i optimalnog meseca za uspostavljanje aseptičnih kultura sorti maline", pages = "480-470", number = "2", volume = "25", doi = "https://doi.org/10.5513/JCEA01/25.2.4201" }
Andjelic, T., Vujović, T., Jevremović, D., Tomić, J.,& Radivojević, D.. (2024). Comparative study of different surface sterilization treatments and optimal month for establishment of aseptic cultures of raspberry cultivars. in Journal of Central European Agriculture Agricultural University Plovdiv, Bulgaria; University of Zagreb, Faculty of Agriculture, Croatia; University of Osijek, Faculty of Agrobiotechnical Sciences, Croatia; Institute of Agriculture and Tourism, Poreč, Croatia; University of South Bohemia, Faculty of Agriculture and Technology, Czech Republic; Hungarian University of Agriculture and Life Sciences, Georgikon Campus, Hungary; Bydgoszcz University of Science and Technology, Poland; University of Agricultural Sciences and Veterinary Medicine, Romania; University of Kragujevac, Faculty of Agronomy Čačak, Serbia; Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources, Slovakia; Agricultural Institute of Slovenia, Slovenia., 25(2), 470-480. https://doi.org/https://doi.org/10.5513/JCEA01/25.2.4201
Andjelic T, Vujović T, Jevremović D, Tomić J, Radivojević D. Comparative study of different surface sterilization treatments and optimal month for establishment of aseptic cultures of raspberry cultivars. in Journal of Central European Agriculture. 2024;25(2):470-480. doi:https://doi.org/10.5513/JCEA01/25.2.4201 .
Andjelic, Tatjana, Vujović, Tatjana, Jevremović, Darko, Tomić, Jelena, Radivojević, Dragan, "Comparative study of different surface sterilization treatments and optimal month for establishment of aseptic cultures of raspberry cultivars" in Journal of Central European Agriculture, 25, no. 2 (2024):470-480, https://doi.org/https://doi.org/10.5513/JCEA01/25.2.4201 . .