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Transient transformation in apple leaves

HK Hui Kang
TZ Ting-Ting Zhang
LF Lu-Lu Fu
YY Yu-Xin Yao
CY Chun-Xiang You
XW Xiao-Fei Wang
YH Yu-Jin Hao
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The 35S::MdCIP1-OX (pRI101-MdCIP1), 35S::MdCOP1-anti (pRI101-asMdCOP1) and EV (pRI101) plasmids were transformed into Agrobacterium tumefaciens strain GV3101. The leaves cut from the 20-day apple culture seedlings were incubated with Agrobacterium, and vacuumed twice for 20 min. The transiently transformed apple leaves were treated on low-nitrogen (0.2 mmol/L nitrate) medium under strong light supplemented with UV-B for 3–5 d at 17 °C. The leaves were harvested for gene expression analysis and anthocyanin content determination [62]. Each experiment required about 60 leaves, and at least three independent biological repetitions were carried out.

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