Abstract
Establishment of an efficient explants surface disinfection protocol is essential for in vitro cell and tissue culture as well as germplasm conservation, such as the case of Grapevine (Vitis spp.) culture. In this research, different procedures for disinfection and regeneration of field-grown grapevine cv. ‘Flame seedless’ axillary buds were evaluated. The buds were disinfected using either NaOCl or allyl, benzyl, phenyl and 2-phenylethyl isothiocyanates. Two different media for shooting and four media for rooting were tested. Shoot and root development per buds were registered. The best disinfection procedure with 90 % of tissue survival involved shaking for 60 min in a solution containing 20 % Clorox with 50 drops/L Triton® X-100. These tissues showed the potential to regenerate a complete plant. Plant regeneration was conducted using full strength Murashigue and Skoog (MS) medium supplemented with 8 µM benzyl aminopurine for shoot induction and multiplication, whereas rooting was obtained on half strength MS supplemented with 2 mg L−1 of indole-3-butyric acid and 200 mg L−1 of activated charcoal. In this work, it was designed the protocols for obtaining sterile field-grown grapevine buds and in vitro plant development. This methodology showed potential to produce vigorous and healthy plants in 5 weeks for clonal grapevine propagation. Regenerated plants were successfully established in soil.
Highlights
Grape (Vitis vinifera L.) is considered one of the most economically important crops in the world (Wang et al 2004)
That is why some species are maintained in germplasm banks to keep their genetic diversity, which is necessary for plant breeding programs (Schuck et al 2011)
Disinfection assay with the NaOCl solutions The best treatment for tissue disinfection was Treatment 2 (T2)-60 with 19.8 % of buds without contamination, and it was statistically significant compared with the control (P < 0.05)
Summary
Grape (Vitis vinifera L.) is considered one of the most economically important crops in the world (Wang et al 2004). That is why some species are maintained in germplasm banks to keep their genetic diversity, which is necessary for plant breeding programs (Schuck et al 2011). It is hard to conserve woody plants in gene banks. The genetic diversity of perennial plants, including grapevines, is usually maintained in field gene banks (Santana et al 2008; Leão and Motoike 2011). These collections are constantly in danger due to exposure to the environment; the conservation of these species requires the development of efficient and cost-effective ex situ protocols, which can be complemented with in situ preservation programs (Touchell et al 2002)
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