Acquity uplc h class xevo g2 xs

The germinated seeds were treated with lovastatin, KT or distilled water as a control. After 4 days of treatments, the seminal roots of rice seedlings were collected and frozen at –80 °C until use. Extraction, purification and quantification of CKs were performed according to the method described previously [52 (link)]. Each sample was ground to a fine power in liquid nitrogen, weighed (~100 mg for each sample) and put into a 1.5-mL tube, mixed with 750 μL cold extraction buffer (methanol:water:acetic acid, 80:19:1, v/v/v), shaken on a shaking bed for 16 h at 4 °C in dark, and then centrifuged at 13,000 rpm for 15 min at 4 °C. The supernatant was transferred to a new 1.5-mL tube and the pellet was remixed with 400 μL extraction buffer, shaken for 4 h at 4 °C, and centrifuged. The two supernatants were combined and filtered using a syringe-facilitated 13-mm diameter nylon filter with pore size 0.22 μm. The filtrate was dried by evaporation under the flow of nitrogen gas at room temperature, and then dissolved in 200 μL methanol. Subsequently, the predominant CKs (Z, DZ, and iP) found in higher plants, and KT was quantified using an acquity UPLC H-Class Xevo G2-XS (Waters Corporation, Milford, MA, USA). Data are means ± standard error (SE) of three independent biological replicates.