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- Author or Editor: Yongjian Chang x
Cold hardiness and cryogenic survival of micropropagated pear (Pyrus cordata Desv.) shoots were evaluated after pretreatments with ABA and sucrose. Shoot cold hardiness increased by 3 °C, and cryopreserved shoot tip growth increased by 17% after a 4-week 150 μm ABA pretreatment. Low temperature (LT) pretreatments improved the recovery of cryopreserved P. cordata shoot tips. Six to 10 weeks of LT were required for reaching high cryopreservation recovery. ABA and LT treatments produced significant synergistic effects on both cold hardiness and cryopreservation recovery. ABA shortened the LT requirement for high cryopreservation growth from 10 to 2 weeks. The optimal treatment for recovery of cryopreserved shoot tips was a 3 week culture on 50 μm ABA followed by 2 weeks of LT, while the maximum cold hardiness (-22.5 °C) was obtained with 150 μm ABA and 2-week LT. A 4 week culture on 150 μm ABA at 25 °C induced dormancy in 74% of shoot tips, but had little effect on cryopreservation growth unless combined with LT. Control and ABA-treated shoot tips, lateral buds, and leaves had similar cold hardiness (-10 to -12 °C), but LT and LT+ABA-treated shoot tips survived the lowest temperatures (-17 to -23 °C), lateral buds next (-15 to -20 °C), and finally leaves (-14 to -18 °C). An increase in the preculture-medium sucrose concentration from 2% to 7% combined with 2-week LT significantly increased cryopreserved shoot tip growth (0% to 75%) and decreased the LT50 from -7.8 to -12.4 °C. The optimal shoot pretreatment for successful recovery of cryopreserved P. cordata shoot tips was a 3 week culture on either 50 μm ABA or 5% to 7% sucrose medium followed by 2 weeks of LT, and increased shoot tip growth from zero to >70%. Chemical name used: abscisic acid (ABA).
‘Hansen 536’ (Prunus dulcis × 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 µm 6-benzylamino-purine (BAP) and 0.98 to 3.94 µm 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 β-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’.