Transient Transformation of Eutrema salsugineum

Eutrema salsugineum Shandong ecotype was grown on soil at 21°C in long day conditions (16 hr light, 8 hr dark). Plant transgenesis was conducted using the floral dip method (Clough and Bent, 1998 (link)). The pEarleyGate 302 vector containing genomic sequence of Arabidopsis thaliana CMT3, including the native promoter, published by Du et al. (2012) (link) was transformed into Agrobacterium tumefaciens strain C58C1. Bacteria were grown for 2 days at 30°C in 200 ml cultures containing gentamicin (25 µg/mL), kanamycin (50 µg/mL), and rifampicin (50 µg/mL) and pelleted by centrifugation at 4°C. Bacterial pellets were resuspended in 5% sucrose and 0.03% Silwet L-77 (Phyto Technology Laboratories) and used to dip open E. salsugineum inflorescences. Transgenic plants were selected for using Finale (BASTA, Bayer).