Figure S1. Leaf phenotype and inter-nodal distance of transgenic Camelina plants. (a) Length of
first leaf (2-week-old plant), (b) width of first leaf (2-week-old plant) and (c) internodal distance
between 3rd and 4th node (5-week-old plant) of CaMV35:AGG3 lines compared to empty vector
lines (EV). Data presented are mean value of 24 plants of each line and error bars represent
standard errors (±SE). Significant difference at *P < 0.05 and ** P < 0.005, respectively
(Student’s t-test).
Figure S2. Vegetative growth parameters of CaMV35:AGG3 overexpression lines. (a) Height of
4-week-old plants, (b) height of mature plants (10-week-old) and (c) number of branches at 5week-old plant. Data presented are mean value of 24 plants of each line and error bars represent
standard errors (±SE). Significant difference at *P < 0.05 and ** P < 0.005, respectively
(Student’s t-test).
Figure S3. Flower-related growth
parameters
of
CaMV35:AGG3
overexpression lines. (a) First flower
initiation day, (b) number of open
flowers at 5 weeks-old plant. Data
presented are mean value of 24 plants
of each line. Flower size as measured
by (c) sepal length, (d) petal length and
(e) petal width of EV and
CaMV35:AGG3 overexpression lines.
Sepal length, petal length and petal
width were measured from mature
flower. The flowers were visualized by
light microscope and picture of
individual parts was taken for
measurement by Image J.
Data
presented are mean values of 12 plants
of each transgenic line. Five mature
flowers were measured from each
plant. Error bars represent standard
errors (±SE) and significant difference
at *P < 0.05 and ** P < 0.005,
respectively
(Student’s
t-test).
Figure S4. Seed number per fruit in overexpression lines. Seed number per fruit was measured
in (a) CaMV:AGG3 and (b) Glycinin:AGG3 transgenic lines and compared with their
corresponding empty vector lines (EV). Data presented are mean value of 24 plants of each line
and ~10 mature fruits were measured from each plant. Error bars represent standard errors (±SE)
and significant difference at *P < 0.05 (Student’s t-test).
Figure S5. Seed viability-related traits of transgenic Camelina plants. (a) Percentage of seed
germination (24 h after transfer of 0.5X MS media containing plates to the growth chamber) and
(b) seed moisture content in CaMV:AGG3 and Glycinin:AGG3 overexpression lines along with
their corresponding empty vector control seeds (EV). Error bars represent standard errors (±SE)
and significant difference at *P < 0.05 (Student’s t-test).
Figure S6. Increased cell division in CaMV:AGG3 lines. Three-day-old plant roots grown on
0.5X MS media were cut near the root tip region and stained with 1X propidium iodide for 15
minutes. The peripheral cellular arrangement was imaged with the Zeiss LSM 510 laser scanning
confocal microscope (Carl Zeiss).
Figure S7. Cell sizes of EV and overexpression lies. Images show comparable cell sizes of 7day-old root, 10-day-old leaf and fully expanded flower petals of empty vector (EV) and
representative CaMV:AGG3 overexpression (OE) line.
Figure S8. ABA hyposensitive seed germination of CaMV:AGG3 overexpression lines.
Percentage of seed germination of empty vector (EV) containing lines was compared with
CaMV:AGG3 overexpression lines on 0.5X MS with or without 1% sucrose in presence of 3 and
5 µM ABA. After 48 h of stratification seeds were transferred to growth chamber and
germination rate was calculated after 36 h, 48 h and 60 h. Panels (a) to (c) represent data
obtained without sucrose in the media and panels (d) and (e) show data obtained in the presence
of 1% sucrose in the media. All seeds of each genotype had germinated fully at 60 h time point
in the presence of sucrose in the media. The experiment was repeated three times and data were
averaged, n=60 per line for each experiment. Error bars represent standard errors (±SE) and
significant difference at *P < 0.05 (Student’s t-test).
Figure S9. Seed germination of Glycinin:AGG3 overexpression lines are ABA hyposensitive in
absence of sucrose. Seeds of empty vector (EV) and Glycinin:AGG3 lines were germinated on
0.5X MS (a) without 1% sucrose and (b) with 1% sucrose in presence of 3 and 5 µM ABA. After
48 h stratification seeds were transferred to growth chamber and percentage of seed germination
rate was calculated after 24 h. The experiment was repeated three times and data were averaged,
n=60 per line for each experiment. Error bars represent standard errors (±SE) and significant
difference at *P < 0.05 (Student’s t-test).
Figure S10. ABA hyposensitive seed germination of Glycinin:AGG3 overexpression lines.
Percentage of seed germination of empty vector (EV) containing lines was compared with
Glycinin: AGG3 overexpression lines on 0.5X MS with or without 1% sucrose in presence of 3
and 5 µM ABA. After 48 h of stratification seeds were transferred to growth chamber and
germination rate was calculated after 36 h, 48 h and 60 h. Panels (a) to (c) represent data
obtained without sucrose in the media and panels (d) and (e) show data obtained in the presence
of 1% sucrose in the media. All seeds of each genotype had germinated fully at 60 h time point
in the presence of sucrose in the media. The experiment was repeated three times and data were
averaged, n=60 per line for each experiment. Error bars represent standard errors (±SE) and
significant difference at *P < 0.05 (Student’s t-test).
Figure S11. High sucrose and NaCl hyposensitivity of CaMV:AGG3 plants. (a) Seeds from EV
and CaMV:AGG3 plants were germinated on 0.5X MS in presence of either 1% sucrose (control)
or 0.4M sucrose. Primary root length was measured from transgenic lines after 4 days of vertical
growth. (b) Seeds from EV and CaMV:AGG3 lines were germinated on 0.5X MS, 1% sucrose
and in presence of 100 mM NaCl and primary root length was measured after 5 days. (c) Seeds
from EV and CaMV35S:AtAGG3 lines were germinated on 0.1X MS and 0.25X MS without
sucrose in presence of 100 mM NaCl and primary root length was measured after 5 days. All
experiments were repeated three times and data were averaged, n=30 per line for each
experiment. Error bars represent standard errors (±SE) and significant difference at **P < 0.05
and *P < 0.01 (Student’s t-test).