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. 2016 Jul 29;172(2):1221–1236. doi: 10.1104/pp.16.00536

Figure 1.

Figure 1.

CI and CII sHSPs are important for basal and acquired thermotolerance. A, Two independent CI and CII RNAi lines and two independent OE lines display corresponding sHSP protein levels. Total protein extracts were obtained from 2.5-d-old wild-type (WT), vector, hsp101, CI or CII OE, and CI or CII RNAi seedlings maintained at 22°C or treated at 38°C for 90 min and allowed to recover at 22°C for 2 h. Samples were separated using SDS-PAGE and analyzed by immunoblotting using the polyclonal antibodies indicated. Staining with Coomassie Brilliant Blue (CBB) was used as a loading control. B, CI and CII sHSPs are important for basal thermotolerance in Arabidopsis seedlings. Dark-grown seedlings (2.5-d) were maintained at 22°C or heat stressed at 43°C for 90 min in the dark, followed by recovery for 24 h at 22°C in illuminated conditions, at which time the percentage of green cotyledons was scored. Results are from a single experiment (n = 18 per line) and are consistent with observations from three experiments. Error bars represent sd. C, CI and CII sHSPs are important for acquired thermotolerance. Soil-grown, 10-d-old wild type, hsp101, vector, and CI and CII RNAi plants were pretreated at 38°C for 90 min, recovered for 2 h, and exposed to 45°C for 10 h. The development of the plants was monitored 5, 10, and 22 d after the heat stress. D, Constitutive expression of CI and CII sHSPs increases basal thermotolerance. Dark-grown seedlings (2.5-d) were maintained at 22°C or heat stressed at 43°C for 105 min in the dark, followed by recovery for 24 h at 22°C in illuminated conditions, at which time the percentage of green cotyledons was scored. Error bars represent sd (n = 24 per line). E, Overexpressing CI and CII sHSPs enhance acquired thermotolerance. The wild type, vector, hsp101, two independent OE lines of CI or CII sHSPs, and an HSP101 OE line were grown on plates for 10 d. Plants were acclimated at 38°C, recovered for 2 h, and exposed to 45°C for 90 min. The percentage of surviving seedlings was determined on day 7. Error bars represent sd (n = 60 per line).