P-GSE40140-4 - P-GSE40140-4

growth protocol
Transgenic lines expressing c-myc tagged WRKY22 in wrky22-ko2 background plants were generated using a T-DNA insertion containing 1000 bp of upstream promoter sequence and WRKY22 full-length genomic DNA in frame with 18 copies of c-myc sequence at 3' end in a pMDC110 derived binary vector. The recombinant plasmid was then mobilized into Agrobacterium tumefaciens strain GV3101 and the resulting GV3101 strain was used to transform wrky22-ko2 plants through floral dipping. Seeds were sown on plates with 0.55% of phytagel (Sigma-Aldrich, St. Louis, MO, USA) in half-strength MS medium (Duchefa Biochemie BV, Haarlem, Netherlands) containing 0.5% sucrose at pH 5.7, and kept at 4oC in the dark for 3 days to achieve uniform germination. The plates were then transferred to a growth chamber and placed vertically, and plants were grown at 22ºC under a 16-hour light (81 μmol s-1 m-2)/8-hour dark cycle. 5-day-old seedlings were transplanted onto fresh plates and the plates were placed vertically to prevent roots from growing into medium. The transplanted seedlings were grown in the growth chamber until they were 14-day-old.
Experiment E-GEOD-40140