Fig. 4.

Phenotypes of the overexpression and RNAi lines of CHLI gene. a Test of the expression levels of CHLI (including CHLI1 and CHLI2) gene in the transgenic lines. Top panel real-time PCR data. Bottom panel immunoblotting data with Actin as a loading control. Col, wild-type Col-0 plants; IR1, IR3, IR4 and IR6, CHLI-RNAi lines 1, 3, 4 and 6, respectively; IOE3, IOE6 and IOE7, CHLI1-overexpression lines 3, 6 and 7, respectively; cs, a T-DNA insertion line of the CHLI1 gene. b Seed germination rate of the wild-type plants (Col) and different transgenic lines as described in (a) in the ABA-free medium (0 μM ABA) and ABA-containing medium (0.5, 1 and 2 μM) 60 h after stratification. Each value is the mean ± SE of five independent biological determinations and different letters indicate significant differences at P < 0.05 (Duncan’s multiple range test) when comparing values within the same ABA concentration. c–e, Early seedling growth of the wild-type plants (Col), cch and cs mutants and different transgenic lines [IR3 and IR6, (d); IOE3, IOE6 and IOE7, (e)] as described in (a) in the ABA-free medium (0 μM ABA) and ABA-containing medium [0.8 μM, (d) and (e); 1 μM, (c)] 12 d after stratification. The experiments were repeated independently five times with the same results. f ABA-induced stomatal closure (top panel) and ABA-inhibited stomatal opening (bottom panel) for wild-type Col, ch1-3 and cs mutants and different transgenic lines (IR3, IR6, IOE6 and IOE7) as described in (a) in the ABA-free medium (0 μM ABA) and ABA-containing medium (20 μM). Values are the mean ± SE from three independent experiments and different letters indicate significant differences at P < 0.05 (Duncan’s multiple range test) when comparing values within the same ABA concentration. n = 60 apertures per experiment. g Whole-plant status in the water loss assays. Intact plants were well watered (Control) or drought stressed (Drought) by withholding water for 15 d for assaying water loss of the two mutants, ch1-3 and cs, and a CHLI-RNAi line IR3 in comparison with wild-type Col. The entire experiment was replicated three times with similar results