A
16
Starch
Carbohydrate content (mg/g FW)
14
WSP
12
10
8
6
4
2
0
WT
isa1- isa1isa2- AtISA1
(P6C3)
isa1- host
AtISA1 AtISA1
(P1D1) (P1A4)
isa1- isa2- host
ZmISA1 ZmISA1 ZmISA1 ZmISA1 ZmISA1 ZmISA1 ZmISA1
(P7E5) (P7E4) (P2D7) (P4F8) (P4F1) (P4F5) (P4D3)
B
WT
isa1- isa2-
P6C3
P1D1
P1A4
P7E5
P7E4
P2D7
P4F8
P4F1
P4F5
P4D3
Scale bars = 10 µm
Supplemental Figure S5:
Panel A: starch and WSP contents determined in host and transgenic lines (as
indicated in the figure). Contents are expressed in mg of carbohydrate per g
of fresh weight (FW) and are the means of three independent biological
replicates. Black and grey vertical bars indicate starch and WSP content
respectively. Vertical lines on bars stand for standard error. Typically no or
very low amount of WSP were detected for WT (Wild Type) and transgenic
lines expressing AtISA1 or ZmISA1 protein.
Panel B: scanning electron microscopy images of purified starch granules
(host and transgenic lines are indicated). The starch samples were prepared
from Arabidopsis leaves harvested at the end of the illuminated period (16h
light / 8h dark). Scale bars = 10µm.
Note that some transformants were not complementing although they were
hygromycin resistant: P7E5 (isa1- exp. ZmISA1) and P4F1 (isa1- isa2- exp.
ZmISA1). Rearrangement of the transgene may have occurred during
integration into the nuclear genome of the plant leading to an alteration of
the ZmISA1 open reading frame and consequently of its expression.