Western blot analysis
Plants were grown on agar plates for ten days. Protein-extracts from whole seedlings were prepared as described by Novák et al.(2018). Proteins were separated on 10% SDS-PAGE and blotted onto nitrocellulose membranes by wet transfer. Membranes were blocked in 5% low fat milk in TBS-T for 1 h, and probed with 1:2,000 anti-PLDα1/2 (AS12 2364, Agrisera, Sweden) in 3% low fat milk in TBS-T for 1 h as well as 1:5,000 goat anti-rabbit (Bethyl) in 5% low fat milk in TBS-T for 1 h. Precision plus protein WesternC standard (Bio-Rad) was used to estimate molecular weights, and this lane was separated from the membrane after blotting and incubated separately in Blocking reagent (Qiagen) in TBS-T for 1 h, followed by 1:10,000 Precision Protein™ StrepTactin-HRP Conjugate (Biorad) for 1 h. For loading control, the membrane was stained with Novex reversible membrane protein stain (Invitrogen) according to manufacturer´s instructions.