Figure 1.
Bioinformatic analysis of NAC protein sequences.
(A) Phylogram was derived for rice, sorghum and maize NAC protein sequences which clustered with EcNAC1 using the program ClustalW. (B) Prediction of putative motifs shared among these protein sequences using MEME program. Numbered boxes represent different putative motifs (annotations of these motifs in EcNAC1 are listed in Table S3).
Figure 2.
Quantitative expression of EcNAC1 gene in response to water-deficit and salt stress.
(A) Adapting a gradual stress imposition protocol, 25-day-old finger millet seedlings were subjected to water-deficit stress following gravimetric approach. (B) NaCl treatment was given to seedlings 3-days after germination. Accumulation of EcNAC1 transcripts was determined by qRT-PCR with 2 µg of total RNA. The Actin gene was used as normalizer.
Figure 3.
Short-term osmotic stress response of EcNAC1 transgenic tobacco plants.
15-day-old T1 transgenic seedlings selected on hygromycin were inter-planted with wild-type seedlings on MS medium supplemented with 5% PEG and observations were taken after seven days of stress treatment. (A) Phenotype, (B) Reduction in fresh weights over control, and (C) Root elongation of transgenic tobacco plants under stress. Each bar value represents the mean ± sd (n = 12) of triplicate experiments (student’s t test; *P<0.05 versus wild-type).
Figure 4.
Short-term salt stress response of EcNAC1 transgenic tobacco plants.
15-day-old T1 seedlings selected on hygromycin were inter-planted with wild-type seedlings on MS medium supplemented with 200 mM NaCl, and the observations were taken after seven days. (A) Phenotype, (B) Reduction in fresh weights over control and (C) Root elongation of transgenic tobacco plants under salt stress. Each bar value represents the mean ± sd (n = 6) of triplicate experiments (student’s t test; *P<0.05 versus wild-type).
Figure 5.
ROS scavenging activity of transgenic plants under water-deficit stress.
15-day-old T1 seedlings were transferred to pots and allowed to establish for 20 days. Water-deficit stress was imposed by gravimetric approach and plants were maintained at 35% FC for 10 days. (A) O2.−, (B) H2O2, and (C).OH radical content were quantified by XTT, scopoletin, and 2-deoxy-D-ribose assay, respectively. Each bar value represents the mean ± sd of triplicate experiments (student’s t test; *P<0.05 versus wild-type).
Figure 6.
Lipid peroxidation in wild-type and transgenic plants under water-deficit stress.
In water-deficit stressed plants MDA accumulation was measured in terms of μM/g fresh weight (FW). Each bar value represents the mean ± sd of triplicate experiments (student’s t test; *P<0.05 versus wild-type).
Figure 7.
Recovery response of transgenic plants subjected to water-deficit stress.
At the end of water-deficit stress period plants were re-watered and observations were taken after 15-days. (A) Recovery growth and (B) Phenotype of wild-type and transgenic plants. Each bar value represents the mean ± sd of triplicate experiments (student’s t test; *P<0.05 versus wild-type).