Abstract
In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.
Highlights
The genetic improvement of woody fruit species, including peach, through biotechnological approaches is often limited by the lack of tissue culture systems that allow the delivery of desired traits in the plant host genome and by the subsequent regeneration of the selected transgenic lines
One of the most important factors affecting plant tissue regeneration is the type and concentrations of plant growth regulators (PGRs) added to the regeneration medium
The highest regeneration efficiency (53%, with mean number of shoots per explant of 0.77 ± 0.08) was obtained when Woody Plant Medium (WPM) basal medium was enriched with 15.5 μM BAP (WPM 11), albeit good regeneration rates were obtained when explants were placed on media supplemented with BAP in combination with naphtalene acetic acid (NAA)
Summary
The genetic improvement of woody fruit species, including peach, through biotechnological approaches is often limited by the lack of tissue culture systems that allow the delivery of desired traits in the plant host genome and by the subsequent regeneration of the selected transgenic lines. Peach is recognized as one of the most recalcitrant species in terms of in vitro organogenesis [2], and only a few studies report the establishment of efficient and reproducible regeneration protocols starting from different explants of peach rootstocks and cultivars. The use of adult tissues is highly recommended to preserve desirable traits of peach selected clones [3], adventitious shoots regeneration starting from seed-derived material has been the main successful approach used in the past for this species [4,5,6,7,8]. Gentile et al (2002) described the only study reporting in vitro shoot organogenesis from leaves of Prunus persica L. cultivars
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