Garlic is affected by various pathogens as in particular some viruses cause significant reduction of plant growth and bulb weight. In a three-years investigation, the effects of main viruses [Garlic common latent virus (GCLV), Shallot latent virus (SLV), Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV)] affecting quality and production was evaluated in Molise region. Pathogenfree plants from 24 local garlic ecotypes, were obtained by a shoot tip in vitro culture protocol including two phases: i) a laboratory pre-screen phase; ii) a subsequent open field post-screen phase. Virus-infected garlic cloves were subjected to thermotherapy (35°C for 15 days) and in vitro plant tissue cultivation followed by RTPCR diagnosis for the presence of viruses. In the pre-screen phase, after the thermotherapy, meristematic tips from garlic cloves were isolated and transferred to nutrient tubes. Shoot growth induction was obtained on a suitable media (Gamborg’s B5, supplemented with sucrose and plant growth regulators). After shoot formation, microplants with microbulb at the base were transferred to a medium inducing development. Total nucleic acid extracts from derived shoots were subjected to RT-PCR analysis, and virus-free samples were further micropropagated. Regenerated garlic plants were again subjected to RT-PCR analysis to confirm the absence of virus. Healthy garlic plants were transferred to soil and kept in climatic chamber for 15-20 days and gradually adapted to natural conditions and then transplanted in the open field under a suitable anti-insect net. All ecotypes will be periodically subjected to pathogen diagnosis throughout the whole plant cycle.
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