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'Hansen 536' (Prunus dulcis 3 Prunus persica) is an important commercial rootstock for peach and almond. However, susceptibility to wet soil and bacterial canker has limited its use primarily to areas with less annual rainfall. Genetic engineering techniques offer an attractive approach to improve effectively the current problems with this cultivar. To develop an efficient shoot regeneration system from leaf explants, 10 culture media containing Murashige and Skoog (MS) or woody plant medium (WPM) supplemented with different plant growth regulators were evaluated, and adventitious shoot regeneration occurred at frequencies ranging from 0% to 36.1%. Optimal regeneration with a frequency of 32.3% to 36.1% occurred with WPM medium containing 8.88 mM 6-benzylamino-purine (BAP) and 0.98 to 3.94 mM indole-3-butyric acid (IBA). The regenerated shoots had a high rooting ability, and 80% of the in vitro shoots tested rooted and survived after being transplanted to substrate directly. Transient transformation showed an efficient delivery of the b-glucuronidase (GUS) reporter gene (gusA) using all three Agrobacterium tumefaciens strains tested with a concentration of OD600 0.5 to 1.0 for 4 days of cocultivation. The protocols described provide a foundation for further studies to improve shoot regeneration and stable transformation of the important peach and almond rootstock 'Hansen 536'. [ABSTRACT FROM AUTHOR]

The organogenesis potential is different among cultivars and must be optimized for individual genotype. Shoot organogenesis capacity from in vitro leaves and root organogenesis capacity of in vitro shoots in six clonal apple rootstock cultivars were compared. The shoot organogenesis capacity was highly genotype dependent. 'GM256' was found to be the most responsive genotype for shoot regeneration from leaf explants among the cultivars, showing high regeneration percentage on all tested media. The effects of basal medium composition and cytokinins on shoot regeneration were different depending on rootstock genotype. Optimum regeneration occurred on Murashige and Skoog (MS) basal medium for '71-3-150', and optimum regeneration occurred on Quoirin and Lepoivre (QL) basal medium for '60-160' and 'ПB'. Thidiazuron (TDZ) was more effective than 6-benzylaminopurine (BA) for Malus prunifolia (Y), whereas TDZ and BA were not significantly different for the other cultivars. All rootstock cultivars showed high root organogenic capacity. The percentage of rooting reached more than 90% and the mean root number per plantlet ranged from three to five. The optimum rooting medium was different for different rootstock cultivars. Optimum root organogenesis occurred on half-strength QL medium for 'GM256' and 'Y', and for 'ПB' and 'JM7' on one-quarter-strength MS medium. [ABSTRACT FROM AUTHOR]

A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawing production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 µM 6-benzylaminopurine (BA), 0.49 µM indole-3-butyric acid (IBA), and 0.29 µM gibberellic acid (GA3). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 µM thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 µM naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 µM BA in combination with 0, 0.54, 1.07, or 5.37 µM NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 µM TDZ plus 0.54 µM NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 µM TDZ plus 1.07 µM NAA. The highest rooting (27%) of adventitious shoots and number of routs per shoot (2.3 ± 0.2) was obtained with 2.5 µM IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 µM IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage. [ABSTRACT FROM AUTHOR]

Plantlets were regenerated from in vitro-grown leaf explants of five genotypes of Liquidambar formosana on WPM basal medium supplemented with different concentrations of TDZ and NAA. With the addition of 0.27 µm NAA, regeneration efficiency was increased by 2- to 4-fold over that with TDZ alone. Lower concentrations of TDZ (0.45-2.27 µM) were beneficial for regenerating shoot clusters. Four genotypes (P2, P6, P9, and P11) showed high regeneration rates (up to 90%), whereas genotype P13 showed a low capability for shoot regeneration on all media tested (<35%). For all five genotypes, the optimum medium for inducing adventitious shoots was WPM supplemented with 1.14 µm TDZ and 0.27 µM NAA, on which regeneration rate ranged from 72.6% to 89.5% and adventitious shoot clusters per regenerating leaf explant ranged from 2.63 to 3.11 in four genotypes (P2, P6, P9, and P11), while for P13, the regeneration rate and number of shoot clusters per regenerating explant were 23% and 1.39, respectively. Transfer of shoot clusters to WPM basal medium containing 0.54 µM NAA, 2.22 µM BA, and 1.44 µM GA3. resulted in shoot elongation. All the elongated shoots were rooted on WPM supplemented with 9.84 µM IBA, and plantlets were transplanted to soil successfully. Chemical names used: 6-benzyladenine (BA), gibberellic acid (GA3), indole-3-butyric acid (IBA), 1-naphthalene acetic acid (NAA), plant growth regulator (PGR), thidiazuron (TDZ), woody plant medium (WPM). [ABSTRACT FROM AUTHOR]

Chokecherry (Prunus virginiana L.) was transformed using Agrobacterium tumefaciens strain EHA105 harboring binary vector pBI121 carrying the neomycin phosphotransferase gene (nptII) and β-glucuronidase (GUS) gene (uidA). Plants were regenerated from the Agrobacterium-infected leaf tissues through organogenesis on woody plant medium (WPM) supplemented with MS (Murashige and Skoog) vitamins, 10 µM 6-benzyladenine (BA), and 250 mg·L-1 cefotaxime plus 500 mg·L-1 carbenicillin plus 15 mg·L-1 kanamycin (CCK15). Transformation was verified with polymerase chain reaction (PCR) and Southern blot analysis. Four of 150 (2.67%) initial explants produced GUS- and PCR-positive shoots. Southern blot analysis confirmed that the transgenes were integrated into the chokecherry genome. Transgenic in vitro shoots were rooted in half-strength MS medium containing 10 µM naphthalene acetic acid. Rooted plants were transferred to potting mix and grown in the greenhouse. This research shows a potential for future improvement of chokecherry and other Prunus species. Chemical names used: 6-benzyladenine (BA), naphthalene acetic acid (NAA), acetosyringone (AS), 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronide cyclohexylammonium (X-Glu), cefotaxime, carbenicillin, kanamycin. [ABSTRACT FROM AUTHOR]