Genetic transformation of A. thaliana and B. napus
The full-length coding sequences of BnGSTU12 was cloned into plant binary expression vector pL-DsRed. The recombinant vector was introduced into Agrobacterium GV3101. A. thalianatransformation was performed by the floral dipping method (Clough and Bent 1998). The B. napus line ZS11 was used as the receptor and transformed using the Agrobacterium-mediated transformation protocol (Yin et al. 2022).
Agrobacterium-mediated transient expression inN. benthamiana leaves and trypan blue staining
The empty vector (EV) as the control and 35S:BnGSTU12-DsRed were transformed into tobacco leaves. Three days after infiltration,N. benthamiana leaves were infected with S. sclerotiorum and lesion area was measured 36 h after inoculation. N. benthamianaleaves was stained by trypan blue. The infected tobacco leaves were decolorized with ethanol, and then submerged to trypan blue solution (0.1% trypan into lactic acid: glycerol: water (1: 1: 1) solution) for 15min. N. benthamianaleaves were cleaned with water and photographed.