Abstract:The effectiveness of in vitro thermotherapy (explants at 35-37°C for 20 days) in obtaining propagating material of apricot (Prunus armeniaca L.) cultivar Bebecou that is free of Plum pox virus (PPV) was compared to a conventional heat treatment technique (potted plants at 30-35°C for 8 weeks). An improved protocol for in vitro thermotherapy was combined with shoot tip culture. The protocol is 5.8 times more effective than conventional thermotherapy while taking half the time and is likely to have a high commer… Show more
“…Although in vitro thermotherapy in combination with shoot tip excision proved to eliminate PPV effectively in apricot cv 'Bebecou' (Koubouris et al, 2007), the survival rate of the explants subjected to in vitro thermotherapy in that study barely reached to 28%. This result is inconsistent with our findings as a decline in survival was observed after prolonged incubation of microshoots to 37°C, implying that woody plants were sensitive to prolonged exposure to extreme high temperatures (Spiegel et al, 1995).…”
Section: Discussionmentioning
confidence: 74%
“…In the case of apricot, thermotherapy was applied by several researchers (i.e., Koubouris et al, 2007;Křižan and Ondrušiková, 2009), however the potential of not only usage of different chemotherapeutic agents for chemotherapy but also application of various cryogenic methods for cryotherapy still needs to be assessed.…”
Plum pox potyvirus (PPV) is the most serious viral disease of stone fruits of Prunus sp. and elimination of the virus could be achieved by using in vitro-applied techniques. Thus, the potential of usage of thermotherapy, chemotherapy and cryotherapy techniques was assessed by in vitro treatment of infected apricot shoot tips. Survival was declined with prolonged application of thermotherapy whereas promising results were scored after chemotherapy and cryotherapy. The highest survival (100%) of the shoot tips was achieved with quercetin containing medium while relatively lower (80%) survival was scored with ribavirin. In addition, inclusion of the 8-azaguanine had found to be phytotoxic as no survival was achieved after treatment. In the case of cryotherapy, two-step freezing technique was applied and 10% of survival was recorded in cryopreserved apricot shoot tips. RT-PCR analysis showed that PPV was eliminated in the survived ones both after chemotherapy and cryotherapy, which revealed that those techniques could be used for in vitro elimination of PPV from infected apricot as well as other stone fruits.
“…Although in vitro thermotherapy in combination with shoot tip excision proved to eliminate PPV effectively in apricot cv 'Bebecou' (Koubouris et al, 2007), the survival rate of the explants subjected to in vitro thermotherapy in that study barely reached to 28%. This result is inconsistent with our findings as a decline in survival was observed after prolonged incubation of microshoots to 37°C, implying that woody plants were sensitive to prolonged exposure to extreme high temperatures (Spiegel et al, 1995).…”
Section: Discussionmentioning
confidence: 74%
“…In the case of apricot, thermotherapy was applied by several researchers (i.e., Koubouris et al, 2007;Křižan and Ondrušiková, 2009), however the potential of not only usage of different chemotherapeutic agents for chemotherapy but also application of various cryogenic methods for cryotherapy still needs to be assessed.…”
Plum pox potyvirus (PPV) is the most serious viral disease of stone fruits of Prunus sp. and elimination of the virus could be achieved by using in vitro-applied techniques. Thus, the potential of usage of thermotherapy, chemotherapy and cryotherapy techniques was assessed by in vitro treatment of infected apricot shoot tips. Survival was declined with prolonged application of thermotherapy whereas promising results were scored after chemotherapy and cryotherapy. The highest survival (100%) of the shoot tips was achieved with quercetin containing medium while relatively lower (80%) survival was scored with ribavirin. In addition, inclusion of the 8-azaguanine had found to be phytotoxic as no survival was achieved after treatment. In the case of cryotherapy, two-step freezing technique was applied and 10% of survival was recorded in cryopreserved apricot shoot tips. RT-PCR analysis showed that PPV was eliminated in the survived ones both after chemotherapy and cryotherapy, which revealed that those techniques could be used for in vitro elimination of PPV from infected apricot as well as other stone fruits.
“…Ram et al [ 33 ] found chemotherapy (30–40 mg L −1 2-thiouracil) followed by thermotherapy (38 °C, 30 days) was most effective in CVB eradication among the five antiviral chemicals tested. 2-thiouracil was also shown to be effective in eradicating LSV, TBV-L and CMV from lily [ 73 ], and PLRV from potato [ 34 ].…”
Section: Key Factors Affecting Success Of Virus Eradicationmentioning
Production of virus-free plants is necessary to control viral diseases, import novel cultivars from other countries, exchange breeding materials between countries or regions and preserve plant germplasm. In vitro techniques represent the most successful approaches for virus eradication. In vitro thermotherapy-based methods, including combining thermotherapy with shoot tip culture, chemotherapy, micrografting or shoot tip cryotherapy, have been successfully established for efficient eradication of various viruses from almost all of the most economically important crops. The present study reviewed recent advances in in vitro thermotherapy-based methods for virus eradication since the twenty-first century. Mechanisms as to why thermotherapy-based methods could efficiently eradicate viruses were discussed. Finally, future prospects were proposed to direct further studies.
“…Thermotherapy is the term used to refer to the application of a high temperature that generates unfavorable conditions for virus multiplication in the cells and effectively decreases the virus load in the shoot tips [9][10][11][12]. Among the established thermotherapybased methods typically used for virus elimination, combined in vitro thermotherapy with shoot tip culture, micrografting, electrotherapy, chemotherapy (especially using ribavirin), and shoot tip cryotherapy has been reported to be efficient for the eradication of different viruses from plants [8,[13][14][15][16].…”
Peach cultivars (’Elberta’, ‘Red Top’, and ’Dixie Red’) were studied for their in vitro adoptability and performance in producing virus-free plantlets. The thermotherapy method with increasing temperatures (25 °C to 37 °C) was applied for the elimination of famous peach infecting plum pox viruses (PPVs) and prunus necrotic ringspot virus (PNRSV), and the DS-ELISA test and RT-PCR technique were performed to confirm the production of virus-free microshoots. The application of 30 mg L−1 of Fe-EDDHA treatment resulted in the best performance for culture establishment of all cultivars, where the best subsequent morphological performance in terms of branch and leaf numbers was recorded for the ’Dixie Red’ cultivar in MS medium, supplemented with 0.5 mgL−1 of gibberellic acid (GA3) and 0.5 mg L−1 of 6-Benzylaminopurine (BAP). At the regeneration stage, the highest (26.96 mm) and lowest (18.43 mm) shoot lengths were obtained from the ’Dixie Red’ cultivar treated with GA3 (2 mg L−1) + thidiazuron (TDZ) (2.5 mg L−1) and the ’Red Top’ cultivar treated with GA3 (1 mg L−1) + TDZ (1 mg L−1), respectively. The leaf numbers were affected by the application of growth regulators, where the ’Elberta’ cultivar under GA3 (2 mg L−1) + TDZ (2.5 mg L−1) treatment showed the highest numbers and the ’Red Top’ cultivar under GA3 (1 mg L−1) + TDZ (1 mg L−1) showed the lowest mean values. The thermotherapy treatment and micropropagation of shoot tips resulted in 100% virus-free plantlets, as confirmed by both applied diagnostic methods. The result of the application of the rooting stage with growth regulators on ’Elberta’ plantlets showed the best performance (90%) in ½ MS medium supplemented with 0.5–1 mg L−1 of IBA, which was significantly higher than the same treatment in MS medium. The obtained results should constitute the basis for further optimization of the multiplication and rooting of virus-free peach plantlets to be served for nurseries and planation orchards.
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