The fragments of these 10 candidate CYP genes were amplified using Phusion® High-Fidelity DNA Polymerase (New England Biolabs, USA) and inserted into the RNAi vector pK7GWIWG2D according to the Gateway procedure (Invitrogen, USA), and the resulting vectors were verified by complete sequencing.
Suspension cells in the logarithmic growth phase were chose and precultured on Murashige and Skoog (MS) solid medium containing 0.5 mg L−1 2,4-D, 0.1 mg L−1 KT, 0.5 mg L−1 IBA and 30 g L−1 sucrose (pH = 5.8) for 7 days. Then, the recombinant plasmid DNA mixed with Au microparticles were transformed into the suspension cells through bombardment using a biolistic gene gun (PDS 1000/He, Bio-Rad). Each transformation was carried out two times. The bombarded suspension cells were cultured for another 7 days before harvesting for qRT-PCR and UPLC analysis73 (link).
Suspension cells in the logarithmic growth phase were chose and precultured on Murashige and Skoog (MS) solid medium containing 0.5 mg L−1 2,4-D, 0.1 mg L−1 KT, 0.5 mg L−1 IBA and 30 g L−1 sucrose (pH = 5.8) for 7 days. Then, the recombinant plasmid DNA mixed with Au microparticles were transformed into the suspension cells through bombardment using a biolistic gene gun (PDS 1000/He, Bio-Rad). Each transformation was carried out two times. The bombarded suspension cells were cultured for another 7 days before harvesting for qRT-PCR and UPLC analysis73 (link).
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