P-TABM-315 - P-TABM-315
Plants were placed with Pieris butterflies for 2-3 h (for Pieris rapae) or 2-6 hours (for Pieris brassicae) to allow time for the butterflies to lay eggs on the leaves. The plants were then placed in a phytotron under the same conditions that the plants were grown previously (8h daylength, 100uE/m/s light intensity, 22C and 70% humidity). Harvests were made at the same time everyday (in relation to the corresponding time for the oviposition) after 24h, 48h, or 72h. For the Pieris rapae eggs, a 5mm cork borer was used to cut a leaf disk surrounding the egg and then the egg was removed. This sample was called the EGG sample. A disk on a distal leaf of the same plant was also cut and referred to as the DISTAL sample. For the Pieris brassicae eggs, for the EGG sample, a scalpel was used to cut the leaf around the eggs, taking the minimum extra tissue possible and the eggs were removed. The scalpel was also used to cut a leaf disk on a distal leaf for the DISTAL sample. After the tissue was cut, it was immediately frozen in liquid nitrogen and stored at -80°C until RNA could be extracted.