Petri César, Scorza Ralph, Srinivasan Chinnathambi
USDA-ARS-Appalachian Fruit Research Station, Kearneysville, WV, USA.
Methods Mol Biol. 2012;847:191-9. doi: 10.1007/978-1-61779-558-9_16.
A high-throughput transformation system for plum has been developed using hypocotyl slices excised from zygotic embryos as the source of explants. The hypocotyl slices are infected in an Agrobacterium tumefaciens suspension and then cocultivated for 3 days in shoot regeneration ¾ MS basal medium supplemented with 9 μM 2,4-dichlorophenoxyacetic acid. Transgenic shoots are regenerated in a medium containing 7.5 μM thidiazuron and elongated in a medium containing 3 μM benzyladenine in the presence of 80 mg/L kanamycin in both media. Transformed shoots are rooted in ½ MS basal medium supplemented with 5 μM NAA and 40 mg/L kanamycin. The transgenic plants are acclimatized in a growth chamber and transferred to a temperature-controlled greenhouse. This protocol has allowed transformation efficiencies up to 42% and enabled the production of self-rooted transgenic plum plants within 6 months of transformation.
已开发出一种用于李子的高通量转化系统，该系统使用从合子胚中切下的下胚轴切片作为外植体来源。将下胚轴切片在根癌农杆菌悬浮液中感染，然后在添加了9μM 2,4 - 二氯苯氧乙酸的芽再生¾ MS基础培养基中共同培养3天。转基因芽在含有7.5μM噻苯隆的培养基中再生，并在两种培养基中均含有80mg/L卡那霉素的情况下，在含有3μM苄基腺嘌呤的培养基中伸长。转化的芽在添加了5μM萘乙酸和40mg/L卡那霉素的½ MS基础培养基中生根。转基因植株在生长室中驯化，然后转移到温度控制的温室中。该方案使转化效率高达42%，并能够在转化后6个月内产生自根转基因李子植株。
