Taurino et al., 2013
SHORT LEGENDS FOR SUPPORTING INFORMATION
Figure S1. Expression analysis of 13-LOX biosynthetic pathway and woundresponsive genes in potato leaves
(a) Total RNA extracted from wounded (L) and systemically induced (S) leaves was
analyzed by northern blot with StLOXH3, StAOC, StOPR3 and StJAR1 specific probes.
(b) Total RNA extracted from wounded (L) and systemically induced (S) leaves was
analyzed by northern blot with StMYC2, StPIN2, StLAP-A and StTD specific probes.
Ethidium bromide staining of rRNA is shown as loading control. The blots presented
here are representative of four independent experiments.
Figure S2. Characterization of single CoAOS1 or CoAOS2 potato plants
(a) Total RNA extracted from wounded (L) and systemically induced (S) leaves from
WT, plants co-suppressed in StAOS1 (CoAOS1-6 and Co-AOS1-12) and plants cosuppressed in StAOS2 (CoAOS2-18 and CoAOS2-19) was analyzed by northern blot
with StAOS1 and StAOS2 specific probes. (b) Total RNA extracted from wounded (L)
and systemically induced (S) leaves was analyzed by northern blot with StPIN2 and
StTD specific probes. Ethidium bromide staining of rRNA is shown as loading control.
The blots presented here are representative of four independent experiments.
Figure S3. Statistical analysis of FTIR data
(a) Distribution of FTIR spectra population from WT (black) and CoAOS1/2-A (blue)
and CoAOS1/2-B (red) cell walls. All treated and normalized spectra were compared
by Principal Component analysis using Win-Das program compressing the data with
the covariance method. We observe the three populations separation in the cluster
plot. (b) Comparison of significant spectra corresponding to the first principal
components of three populations.
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Taurino et al., 2013
Figure S4. Identification of pectin oligosaccharides 868.42 and 842.70
(a) Product ion spectra obtained on a MALDI TOF/TOF (TOF-Q2) instrument for the
parent ion at m/z 868 (tentative structure: [GalA2Ac1Me1Hex1Pent1dHex1+]+).
Mass losses of 146 and 162 Da are indicative respectively of the presence of
rhamnose and hexose. Mass loss of 192 Da from the ion parent indicates that
oligosaccharide includes GalA. The 150 u.m.a cleavage product indicates the
presence of a pentose (Pent+H2O) in the oligosaccharide. Structure corresponds to a
rhamnogalacturonan oligosaccharide with an alternating GalA–Rham backbone where
rhamnose can be substituted with galactose and arabinose in side chain. (b) Product
ion spectra obtained on a MALDI TOF/TOF (TOF-Q2) instrument for the parent ion at
m/z 842 (tentative structure [GalA1Hex4]+). Mass loss of 192 Da from the ion parent
indicates that oligosaccharide includes GalA, a concatenated mass losses of 164 Da
indicates the presence of multiple hexoses.
Figure S5. Oligosaccharide mass profiling of PG digested cell walls
MALDI-TOF/MS analysis of pectic oligosaccharides after digestion of cell walls of WT
(top),
CoAOS1/2-A
(middle)
and
CoAOS1/2-B
(bottom)
leaves
with
endo-
Polygalacturonase.
Figure S6. In situ hybridization control
Non-wounded (left), 4h locally wounded (middle) and systemically induced potato
leaves (right) hybridized with a potato histone 4 (H4) antisense probe. Signal was
detected as a purple precipitate when viewed under the light microscope. Bar: 100
μm.
Table S1. Oxylipin profile upon wounding of transgenic potato plants
specifically depleted of each StAOS
Content of OPDA, JA and JA-Ile in wounded and distal leaves of WT plants, plants cosuppressed in StAOS1 (CoAOS1-6 and Co-AOS1-12) and plants co-suppressed in
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Taurino et al., 2013
StAOS2 (CoAOS2-18 and CoAOS2-19). Leaves were collected at the times indicated.
Each value corresponds to the average and standard deviation of data measurements
from 3 independent biological experiments.
Table S2. Neutral and acidic monosaccharide composition (%mol) by
trimethyl-silyl-oxymes of cell walls obtained from transgenic and wild type
leaves. Data ± SD are the average of five independent experiments. Statistical
analysis of the data was performed using Student´s t test. No significant differences
were found. GlcU, Glucuronic acid; GalU, Galacturonic acid. Nd, not detectable.
Table S3. Primers used in this study or restriction enzyme digestion
approach used to generate specific probes.
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