Mlr 350 h growth chamber

A colony of the two-spotted spider mite T. urticae London strain (Acari: Tetranychidae), provided by Dr. Miodrag Grbic (UWO, Canada), was maintained on beans in a Sanyo MLR-350-H growth chamber at 25°C under a 16 h light/8 h darkness photoperiod. This colony was transferred to barley where it was maintained under the same conditions for more than 30 generations to ensure host adaptation To induce biotic stresses mediated by pest attack, 7-day-old barley plants, wild-type and transgenic lines overexpressing or silencing the HvPap-1 gene, were infested with 20 barley-adapted adults of T. urticae per plant. Seven plants were used per treatment and three independent experiments were performed. Barley plants were confined in independent pots with a plastic cylinder covered on top by nylon nets to avoid dispersion of mites and the same isolation system was applied to control plants. Plants were further incubated at 25°C under a 16 h light/8 h darkness photoperiod. Barley leaf damage was monitored at different time points after spider mite feeding. Leaves were harvested after 7 and 14 days of mite treatment. Samples were imaged and scanned, or frozen into liquid nitrogen and stored at -80°C for further analysis.

Arabidopsis thaliana (L.) Heynh. Columbia-0 and phot1 (SALK_088841) seeds were purchased from the Nottingham Arabidopsis Stock Center (NASC, Nottingham, UK). phot2 seeds were the kind gift of J. Jarillo. The phot1phot2 double mutant was obtained by crossing phot1 and phot2 plants [50 (link)]. Plants were grown in a Sanyo MLR 350-H growth chamber, at 23 °C, 80% humidity, in a 14 h light/10 h dark photoperiod. The chamber was equipped with fluorescent lamps (OSRAM L36W/77 and PHILIPS Master TL-D 36/W/840), yielding an average irradiance of 70 μmol m⁻² s⁻¹ for 5–6 weeks.

A colony of the two-spotted spider mite T. urticae London strain (Acari: Tetranychidae), provided by Dr. Miodrag Grbic (UWO, Canada), was maintained on beans in a Sanyo MLR-350-H growth chamber at 25°C under a 16 h light/8 h darkness photoperiod. This colony was transferred to barley where it was maintained under the same conditions for more than 30 generations to ensure mite adaptation to the host.