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Možnosti využití antivirotik v procesu eliminace virů u česneku kuchyňského
Kudělková, Martina
The dissertation is focused on the effect of three antivirals (acyclovir, rimanatadine or zidovudine) for their potencial use in the process of chemotherapy in the combination of meristem culture method as a possible method for Onion yellow stripe virus, Leek yellow stripe virus, Garlic common laten virus and Shallot latent virus eradication in in vitro conditions. Antivirals were always applied separately into cultivation media and two amounts of antivirals (25 or 50 mg.l-1) were used. The treatment time was two weeks. Meristem culture only was a control method. Murashige and Skood medium was used for all variants. During the first year, young plants of the variety D Alsace Freres cultivated in vitro were treated with chemotherapy. These plants had been treated with the meristem culture only in the past but the treatment had not been successful. The virus presence was detected with the ELISA method. Rimantadine variant in the amount of 50 mg.l-1 was the most effective variant for the monitored virus elimination. More sensitive method reverse transcription polymerase chain reaction (RT-PCR) was optimized for monitored viruses in the next parts of this work. Garlic common latent virus (GCLV) primers were designed as well. During the second year, the chemotherapy was applied on explants with the size 0.8 mm during meristem culture. Explants of the varieties Blanin, Sukoradský and Japo, however, did not regenerate and the chemotherapy could not be evaluated. In the next two years of the work, the chemotherapy was applied on individual cloves of the Unikát variety. Whilst detecting the presence of viruses in plants prior the treatment, it was established that only one plant was free from all the monitored viruses. After treatment, acyclovir variant in amount of 25 mg.l-1 was evaluated as the most effective variant for the virus complex elimination, and also from the economical point of view. Real-Time PCR method for virus detection has been started in the dissertation. The detection of GCLV was optimized from the start. Isolates which had been negative for the GCLV presence after RT-PCR were tested with Real-Time PCR. A higher percentage of GCLV positive plants were detected with Real-Time PCR. Real-Time PCR optimization for other viruses will be the objective of futur experiments.
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