Fig. 3

Over-expression and expression of NoDGTT5 in N. oceanica CCMP1779 under control of the EF promoter (D5oxC and D5oxD) and the native promoter (D5proA and D5proC), respectively. a Growth of wild-type stain (WT), control vector transformants (EV), and lines over-expressing NoDGTT5 under N-replete (N+) and N-depleted (N−) conditions. b Relative expression of NoDGTT5 analyzed by qPCR in transformants growing under N+ and N− conditions. qPCR values were normalized to the ACTIN qPCR values and to the expression level of the wild-type under the respective condition (N+ or N−). Each data point represents the average of three biological replicates. Each sample was analyzed in technical triplicates. Values represent mean ± SD (n = 3) and asterisks indicate P < 0.01. c Relative abundance of TAG in strains used in b. d, e Positional analysis of TAG acyl groups of the WT strain and NoDGTT5 over-expressing line growing in N+ (d) and N− (e) media. Data are presented as mean ± SD (n = 4–6), with asterisks indicating P < 0.05. f Detection of venus fluorescence in over-expressing lines D5oxC and D5oxD by using CLSM. Bars 1.5 μm. g Localization of NoDGTT5-GFP constructs expressed under native promoter in D5proA line; CHL chlorophyll; LDs lipid droplets. Bars 1.5 μm