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Sabbadini S.,Marche Polytechnic University | Cappelletti R.,Marche Polytechnic University | Mezzetti B.,Marche Polytechnic University | Pandolfini T.,University of Verona | And 4 more authors.
Acta Horticulturae | Year: 2016

Prunus, in particular peach, is considered one of the most recalcitrant species for what concerns in vitro regeneration, especially when the starting plant material originates from mature tissues. Furthermore, in terms of peach genetic transformation, few examples exist in literature and most of them demonstrate the difficulty in obtaining stable transformants. The present study describes an efficient method of in vitro regeneration, via organogenesis, starting from meristematic bulk tissues, characterized by a high competence to produce adventitious shoots and useful for genetic transformation of Prunus persica cultivar 'Big Top® Zaitabo∗' and rootstock GF677. This protocol constitutes one of the few examples of peach in vitro regeneration starting from adult tissue present in the literature. It has been used to genetically transform the rootstock GF677 and the cultivar 'Big Top® Zaitabo∗', through the development of two different methods of transformation, one mediated by Agrobacterium tumefaciens, applied on slices from meristematic bulk, and the other by the biolistic technique applied on slices from meristematic bulks and on vegetative apex. The transformation experiments were conducted using hairpin genetic constructs for the induction of Plum pox virus (PPV) resistance through the mechanism of post-transcriptional gene silencing (PTGS). To our knowledge, this is the first example of a successful attempt of a peach rootstock and cultivar stable genetic transformation and it represents a good promise for the future production of silenced GF677 plants to be tested for the capacity to induce PPV resistance also in the grafted scion. Source


Navacchi O.,Vitroplant Italia | Zuccherelli G.,Vitroplant Italia | Mazzucchi U.,University of Bologna
Acta Horticulturae | Year: 2013

In industrial micropropagation laboratories culture substrate microbial contamination is a problem. Visible contamination can be easily recognised and eliminated. In the case of not detectable contamination, however, there is a high risk of transmitting and spreading the microorganisms vertically or horizontally in the laboratory. Thus, it is essential to have strategies for establishing and maintaining aseptic tissue cultures. To ensure that in vitro cultures of the globe artichoke 'Life' are aseptic, we developed a strategy to eliminate a persistent, substrate contaminant bacterium, associated with substrate turbidity that is almost invisible over time, which apparently did not compromise plantlet development, but was resistant to common sanitization measures. Ambivalent media were developed to favour the growth of the plantlets as well as bacteria, so that to make visible their presence. Latently contaminated globe artichoke explants, propagated on standard MS media with BA (0.1 mg/L), were transplanted to standard media modified with the addition of combinations of microbial growth stimulants (yeast extract, malt extract, peptone, casamino acids, proteose peptone 3). The plantlets were subsequently transplanted on the same media three times, about every 15 days. The bacterium, identified as a strain of Bacillus nealsonii or Bacillus benzoevorans, developed visible colonies on all the media, even those with the lowest concentrations of growth promoters. The results of bacterial growth tests on the substrate variants indicate that the modified MS medium, with addition of (0.02%) yeast extract, (0.05%) casamino acids and (0.05%) proteose peptone 3, promotes the growth of the contaminant bacterium, as well as the growth of 'Life' globe artichoke plantlets in vitro. The highest concentrations of growth stimulators were shown to be phytotoxic. Our results show that by using an appropriate ambivalent medium, it is possible to highlight the presence of contaminant bacteria in globe artichoke culture media and facilitate an accurate selection of aseptic jars especially in the early stages following the transplantation of the mother plant tissues. Source


Pirona R.,Parco Tecnologico Padano | Zuccherelli G.,Vitroplant Italia | Capaccio V.,Vitroplant Italia | Ciannamea S.,Parco Tecnologico Padano | And 2 more authors.
Acta Horticulturae | Year: 2013

The globe artichoke is extensively cultivated in the Mediterranean region. Italy is the first country for artichoke production in the world (http://faostat.fao.org). Immature artichoke flowers are usually consumed as fresh, frozen, or canned vegetables. A description is given of the genetic analysis of the 'Apollo' cultivar, using microsatellites (SSRs), to validate the hypothesis of a different genetic background from other commercial cultivars used in this study. Seven microsatellites, each one located on a different chromosome, resulted sufficiently informative to classify the two cultivars developed by Vitroplant ('Apollo' and 'Apollo' mutant) and discriminating them from other commercial cultivars tested in this study. Source

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