Gamborg s b5 salt medium

Tomato ‘Avigail’ )870) was used as the background line for both transgenic root lines: mj-far-1 OE and the control Kan, as described previously [26 (link)]. Both root lines were subcultured on standard-strength Gamborg’s B5 salt medium (Duchefa, Haarlem, The Netherlands), supplemented with 2% (w/v) sucrose and solidified with 0.8% (w/v) Gelrite agar (Duchefa). Roots were subcultured on B5 medium, with one root section per petri dish (Miniplast, Ein Shemer, Israel), and incubated horizontally in a growth chamber at 26°C in the dark for 1 week to allow root branching before nematode inoculation.

Sterilized tomato seedlings were grown at 26 °C under 16 h daylight in sterile plates on standard-strength Gamborg's B5 Salt medium (DUCHEFA, Haarlem, The Netherlands), supplemented with 2% (w/v) sucrose and solidified with 0.8% (w/v) Gelrite agar (DUCHEFA). Roots were subcultured on the same medium with one root section per petri dish (MINIPLAST, Ein Shemer, Israel) and incubated in a growth chamber at 26 °C in the dark for 2 weeks. Protoplasts were released and isolated from about 40 plates of roots using Demidchik and Tester's^{116 (link)} protocol. Fresh protoplasts were incubated with 60,000 freshly hatched sterilized J2 at 26 °C in the dark for 3 h.