title: RNA-Seq of Arabidopsis thaliana and Arabidopsis lyrata plants primed by insect eggs from Pieris brassicae and triggered by insect herbivory
design: RNA-Seq of Arabidopsis thaliana and Arabidopsis lyrata plants primed by insect eggs from Pieris brassicae and triggered by insect herbivory
library_name: LYR_EGG_1_p
library_construction_protocol: Samples were collected in liquid nitrogen until nucleic acid extraction. Arabidopsis thaliana (Columbia-0) and Arabidopsis lyrata ssp. lyrata (RonC) were sown on a 3:1 mixture of soil:sand and stratified for two day at 4°C. The plants grew in climate chambers under short day conditions (10 h/ 14 h light dark cycle, 21°C, 40% relative humidity and light intensity of 100-120 µmol m-2 s-1 light intensity). To treat a plant with P. brassicae eggs we exposed one fully developed, non-senescent leaf (position 17-22) of the Arabidopsis plants to one mated female butterfly. Per leaf the butterfly laid one egg clutch with 30-40 eggs. After six days under short day conditions (10 h/ 14 h light dark cycle, 21°C, 40% relative humidity and light intensity of 100-120 µmol m-2 s-1 light intensity), the eggs were removed using pairs of tweezers and a fine brush.  To treat the plants with larval feeding, we have transferred 10 neonate larvae to the plants. Neonates have developed on Brussel sprouts. One day before hatching, the eggs were removed from the plants and collected in Petri dished where larvae hatched one day later.  The neonate larvae were enclosed in clip cages (2 cm in diameter, 1.7 cm high). After two days feeding on previously egg-laden or egg-free plants, we have determined larval weights and harvested leaf material for transcriptome analyses. The leaf material was flash frozen in liquid N2. To determine larval weights after a 5-day-feeding-period, the larvae could move and feed freely on the entire plant. To prevent larvae from escaping, the plants were enclosed in plexiglas® cylinders (14.5 cm diameter, 15 cm high) with a gauze lids. The citric acid RNA extraction protocol by Oñate Sánchez and Vicente-Carbajosa (2008) was used followed by DNAse treatment with the TURBO DNA Free kit (Ambion). Illumina TruSeq RNA library kit at Macrogen Inc.
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-12653:LYR_EGG_1_p