Dhandapani et al.

Table S1

Prediction of the sub-cellular localization of GhCCL

Chloroplast with score:3.0

Chloroplast Transit peptide 1-9 is found

Chloroplast Transit peptide (cTP) length: 48

Integral Prediction of protein location: Chloroplast with score 8.7

Location weights / LocDB / PotLocDB / Neural Nets / Pentamers / Integral
Nuclear
Plasma membrane
Extracellular
Cytoplasmic
Mitochondrial
Endoplasmic reticulum
Peroxisomal
Golgi
Chloroplast
Vacuolar / 0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0 / 0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0 / 0.01
0.00
0.00
0.00
0.00
0.00
0.00
0.00
2.99
0.00 / 0.40
0.36
0.02
0.68
0.43
0.00
0.08
0.11
2.54
0.39 / 0.00
0.54
0.00
0.00
0.53
0.04
0.00
0.09
8.73
0.08

Table S2

Leaf disc senescence assays of Wild-type and T0 transgenic tobacco in distilled water supplemented with 10% PEG-6000, 200mM each mannitol and NaCl. Percentage of yellow and green leaf disc were scored after 72 hrs of each treatment (n=90 leaf disc for each test)

Transgenic lines / 10% PEG-6000 / 200mM mannitol / 200mM NaCl
Percentage (%) / Percentage (%) / Percentage (%)
Green Leaf Disc / Yellow Leaf Disc / Green Leaf Disc / Yellow Leaf Disc / Green Leaf Disc / Yellow Leaf Disc
WT / 15.56±1.69 / 84.43±1.69 / 17.76±1.69 / 82.23±1.69 / 22.23±3.46 / 77.73±3.46
A3 / 85.56±1.69 / 14.43±1.69 / 86.66±1.11 / 13.30±1.11 / 24.46±3.46 / 75.53±3.46
A6 / 84.46±2.30 / 15.53±2.30 / 86.66±2.21 / 13.33±2.21 / 22.23±2.54 / 77.75±2.54
B6 / 88.86±1.69 / 11.10±1.69 / 76.66±1.11 / 23.33±1.11 / 17.70±2.54 / 82.23±2.54
B20 / 76.66±1.11 / 23.33±1.11 / 81.10±2.80 / 18.90±2.80 / 16.66±1.67 / 83.33±1.67
CD (0.05) / 10.9031 / 10.9031 / 11.9029 / 11.9029 / 9.3980 / 9.3980

Each value represents the mean ± of triplicates (ANOVA test) and means are significantly different from control at P < 0.05 according to Dunnett’s test at each treatment.

Table S3

Seed germination data of wild-type and T1transgenic tobacco in MS agar medium supplemented with 10% PEG-6000, 200mM each mannitol and NaCl. Data were collected 20 days later (n=300 seeds for each test)

Transgenic lines / 10% PEG / 200mM mannitol / 200mM NaCl
WT / 40.0±1.6d / 33.6±1.4c / 25.6±2.0d
A3 / 97.3±0.7a / 89.3±1.3a / 41.3±1.07c
A6 / 95.0±1.4a / 86.3±1.7a / 76.6±1.01a
B6 / 82.6±1.7b / 67.6±2.0b / 53.3±1.57b
B20 / 49.6±1.7c / 66.6±1.4b / 27.6±0.50d
CD (0.05) / 7.5862 / 10.3899 / 6.7100

Each value represents the mean ± of triplicates (ANOVA test) and means are significantlydifferent from control at P < 0.05 according to Dunnett’s test at each treatment

Table S4

Wild-type and T1transgenic tobacco stem portion was cultured on MS medium supplemented with 10% PEG-6000, 200mM each mannitol and NaCl. Fresh weight was taken after 45 days grown plants for each treatment. (Fresh weight calculated by Fresh weight of respective lines under stress x 100/ fresh weight of respective plant with 0 conc. treatment)

Transgenic lines / 10% PEG / 200mM mannitol / 200mM NaCl
WT / 48.5±0.46e / 31.7±1.36d / 40.4±1.07d
A3 / 57.4±0.75d / 51.1±2.08c / 68.5±1.68a
A6 / 65.9±1.33b / 64.4±1.73a / 49.5±1.97c
B6 / 67.8±0.57a / 63.0±0.31a / 57.9±0.23b
B20 / 62.8±0.81c / 59.1±0.91b / 71.7±1.92a
CD (0.05) / 1.6230 / 2.8098 / 3.2110

Each value represents the mean ± of triplicates (ANOVA test) and means are significantly different from control at P < 0.05 according to Dunnett’s test at each treatment

Table S5

Relative water content analysis of wild-type and T1transgenic tobacco plants under glass house condition upon before and after 15 days water withholding.

Transgenic lines / Before water withholding / 15 days after water withholding
WT / 49.7±2.1d / 43.1±0.7d
A3 / 50.1±1.3c / 48.8±0.7c
A6 / 50.3±1.3bc / 49.2±0.6c
B6 / 52.9±1.3ab / 51.2±0.0b
B20 / 55.2±1.42a / 53.8±1.75a
CD (0.05) / 8.0946 / 4.7812

Each value represents the mean ± of triplicates (ANOVA test) and means are significantly different from control at P < 0.05 according to Dunnett’s test at each treatment

Table S6

Total chlorophyll content analysis of wild-type and T1transgenic tobacco plants under glass house condition upon before and after 15 days water withholding

Transgenic lines / Before water withholding / 15 days after water withholding
WT / 3.41±0.017 / 3.04±0.040
A3 / 3.72±0.045 / 3.58±0.037
A6 / 3.65±0.010 / 3.50±0.033
B6 / 3.38±0.025 / 3.34±0.016
B20 / 3.79±0.083 / 3.70±0.068
CD (0.05) / 0.2596 / 0.2207

Each value represents the mean ± of triplicates (ANOVA test) and means are significantly different from control at P < 0.05 according to Dunnett’s test at each treatment