The Agrobacterium single
colony was cultured in 25 ml Luria–Bertani (LB) liquid medium
containing 50 mg/l kanamycin and 50 mg/l rifampycin on a reciprocal
shaker (200 rpm) at 28 ◦C for 16–18 h until OD600nm = 0.5
was reached. The bacteria were collected by centrifugation at
5000rpm for 5min, washed with LB liquid medium and further
with Murashige and Skoog (1962) (MS) medium containing
2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), and then were
resuspended in 25 ml MS medium containing 2.0 mg/l 2,4-D for the
inoculation