Abstract
New Plant Breeding Techniques (NPBTs) protocols have been developed to produce new grape varieties with improved quantitative and qualitative characteristics. Reliable transformation protocols for grapes are based on the generation/induction of embryogenic callus cells that are then transformed. Varieties such as Italia have proven to be very recalcitrant to regeneration via somatic embryogenesis. In this work, the development of a protocol for improved production of embryogenic calluses is described. Two sterilization protocols were tested: (a) a lower active chlorine concentration for a longer time (LS); and (b) a higher chlorine concentration for a shorter time (HS), in combination with the absence or presence of citric acid in the growing substrate in the first growth media. The embryogenic calluses formation in Chardonnay, a cv. with a high embryogenic response, was significantly higher in presence of citric acid in the initial growing substrate regardless of the sterilization protocol. In Aglianico, a cv. with a lower embryogenic response, no significant differences were observed. Instead, in a recalcitrant cv. as Italia, we obtained a 13-fold increase in embryogenic calluses formation performing sterilization of flowers with the HS protocol compared to LS.
Highlights
Is among the major grape producers (13.5% table grape, 86.5% wine grape), with a large vineyard-dedicated surface area [1]
In order to set up an optimal protocol for induction of embryogenic callus from Vitis vinifera varieties, we gathered several culture media from different works [19,25]
To regenerate a new plant with genetic engineering, it is necessary to have tissues with embryogenic competence. These tissues in vines can be obtained from cultures of flowers, ovaries, or anthers collected in a specific and narrow window of time
Summary
Is among the major grape producers (13.5% table grape, 86.5% wine grape), with a large vineyard-dedicated surface area [1]. Italian table grape production has long been dominated by the “Italia” variety, which has large white aromatic but seeded berries. In the last decade, numerous new seedless varieties have been registered in the Italian National Register of Vine Varieties [2]. Most of these varieties have a non-EU origin, which might result in adaptation difficulties in European cultivation areas. The main objectives for the genetic improvement of table grape breeding programs are large seedless berries with appreciated organoleptic characteristics, an extension of the harvest calendar, simplified cultivation management of the vineyard, good productivity, enhancement of nutraceutical composition, improved aptitude for cold storage, suitability for transport, and long shelf-life.
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