RWC was then calculated Selleck NVP-AUY922 as following: RWC (%) = 100 − WL (Gao and Ye 2007). Drought stress was achieved by

the addition of PEG 6000 (PEG-6000, MW 6000; Merck, Darmstadt, Germany) to culture medium. Different concentrations of PEG were added to 100 mL cultures of tested strains to give water potentials of 0, −0.15, −0.30, −1.03, and −1.76 MPa (equivalent to 0, 15, 20, 30, and 40% (w/v) of PEG-6000, respectively) according to the methods of Mohammadkhani and Heidari (2008). Algal cells grown under exponential growth phase were used for this study. The growth rate was calculated on the basis of incubation for 3 d. Chl-a was used as a measure of growth. Ratios of chl-a to dry weight were checked to confirm the suitability of using chl-a as a measure for the studied organisms. Algal cells were harvested by centrifugation (4,000g × 10 min). Dry weight was measured by filtering a 100 mL sample through

pre-weighted 0.45 μm Whatman GF/C filters and drying the cell mass at 70°C for 24 h (Fan et al. 1994). Chl-a and carotenoids from samples were extracted in 90% acetone and the abundance of the pigments was determined from absorbance at 450, 645, and 663 nm, using the methods of Inskeep and Bloom (1985). Specific growth rate (μ, d−1) was determined using the equation suggested by Myers and Kratz (1955) as follows: Y-27632 purchase μ = ln (N1/N0)/(T1 − T0), where N1 and N0 are the final and initial concentrations of chl-a at time T1 and T0, respectively. Megestrol Acetate The growth rate per day was used as a

basis of comparison in this study. The methods of Tandeau de Marsac & Houmard (1988) were used to estimate the concentration of PC and APC in cyanobacterial cells. The cells were soaked in 20 mM sodium acetate buffer (pH 5.5) and subjected to disruption with a bead beater (Biospec, Bartlesville, OK, USA). After centrifugation (10,000g × 10 min), 1% (w/v) streptomycin sulfate was mixed with the supernatant for 30 min at 4°C. Samples were centrifuged (10,000g × 10 min) at 4°C and the absorbance at 620 and 650 nm of the supernatant was measured by a Beckman Coulter DU800 spectrophotometer (Brea, CA, USA). Lipid peroxidation was estimated by measuring the formation of MDA with TBA, according to the methods of Heath and Packer (1968) modified by De Vos et al. (1989). The samples were suspended in 1 mL H2O (bi-distillated) and mixed with an equal volume of sea sand. Subsequently, the cells were disrupted with a bead beater and centrifuged to separate the aqueous extracts from debris. Then, 1 mL of 0.25% TBA dissolved in 10% trichloroacetic acid was added and incubated at 95°C–100°C for 30 min, then chilled on ice. Samples were centrifuged, and MDA was determined by subtracting absorbance of the supernatant at 600 nm from that at 532 nm. To calculate the concentration, an absorbance coefficient of 155 mM−1 · cm−1 was used (Kwon et al. 1965).