Evaluations carried out by Universidade de Tras-os-Montes e Alto Douro, Portugal as part of the RESGEN project
Glucosinolates quantification
Seedling plants were grown in growth cabinets under controlled conditions to evaluate the glucosinolate content at 3-true leaf stage. In this experiment 3 replicates with 5 plants per replicate were used, all used for sampling. From sowing to emergence light was set at 400 µmoles.m-2.s-1 and at constant day/night temperature of 22ºC. After emergence to harvest date light was kept at as previous and temperature was changed to a regime of 20/16ºC day/night, according to plant requirements. Seedlings were harvested 14 days after sowing.
Mature plants were grown in the field. Sowing was done in 2 different periods in peat blocks (6x6x6 cm) and seedlings were transplanted into field at the 3-true leaf stage and set in raise beds in 35 x 60 cm spacing. The experiment was set in a complete randomized block design with 3 replicates with 6 plants each accession. Harvest was done when plants reached the maturity stage, per replicate collecting from 3 plants different kind of plant parts (inflorescences, leaves and roots), according to plant species.
Following the harvest the samples were homogenised in liquid nitrogen and an aliquot was freeze-dried for further analysis by HPLC.