Figure S1 Enod11 Mtc27 MtGshs cDNA gDNA S. meliloti DPI --++ - + + - Supporting Information Fig. S1....
-
Upload
edmund-blankenship -
Category
Documents
-
view
220 -
download
0
Transcript of Figure S1 Enod11 Mtc27 MtGshs cDNA gDNA S. meliloti DPI --++ - + + - Supporting Information Fig. S1....
Figure S1
Enod11
Mtc27
MtGshs cDNA
gDNA
S. meliloti
DPI
- - ++
- + +-
Supporting Information Fig. S1. Validation of the selected biological conditions for MtEnod11 gene expression and ROS levels. (a) After root treatment (see methods), the level of expression of Enod11 and Mtc27 in the biological samples used for transcriptome analysis was evaluated by RT-PCR. We checked that there was no genomic DNA in RNA samples with primers spanning the intron of MtGshs1. Representative colorimetric detection of the superoxide anion (NBT; b) and hydrogen peroxide (DAB; c) in root hairs. Scale bar = 50 µm. n > 10
(a)
(c)(b)
R2 = 0.9348
-10
-8
-6
-4
-2
0
2
4
6
8
10
-10 -8 -6 -4 -2 0 2 4 6 8 10
Figure S2
Supporting Information Fig. 2. Comparison of expression ratios of probesets in RT-qPCR and Affymetrix gene array analyses. Analysis, RT-qPCR, of the differential accumulation of transcripts, initially identified by Affymetrix GeneChip analysis (Table S2), on two new biological replicates (data from Table 1).
Fol
d ch
ange
Log
2 R
T-q
PC
R
Fold change Log2 Affymetrix
Figure S3
Con
trol
HyP
er
66 kD
45 kD
(a)
0
25
50
75
100
450 500 550 600 650
Wavelength (nm)
Arbi
trary
uni
ts
(b)
Supporting Information Fig. 3. HyPer protein production in composite plants of Medicago truncatula. Western-blot analysis of HyPer in root cultures expressing a control or HyPer construct. (b) Representative emission spectra after 405-nm excitation () and 488-nm excitation (—).
Rip1 bHlhSpk1Spk2AblL Srl1 Nin Pri1
10
Fo
ld c
ha
ng
e
Figure S4
Supporting Information Fig. 4. Relative expression of selected genes in untreated Medicago truncatula roots. The relative expression level of the selected genes was evaluated before H2O2 treatment in two biological replicates. The baseline corresponds to MtHap2-1 expression, the gene least strongly expressed in our conditions.
Figure S5
Control amiSpk1
(a) (b)
(c) (d)
Supporting Information Fig. 5. Phenotypes of infection threads in amiRNA MtSpk1 plants. Composite plants expressing either an empty vector (control; a, c) or the amiRNA against MtSpk1 (amiSpk1; b, d) were inoculated with LacZ (a, b) and DsRed (c, d) S. meliloti strains. IT were observed 4 dai (a, b) and 10 dai (c, d), as described in Figure 5. n > 10. Arrows show ITs. Scale bar = 100 µm.
Figure S6
Supporting Information Fig. 6. Subcellular distribution of MtSPK1. Composite plants expressing a cytosolic GFP (a), a BUBR1::GFP (b; nuclear protein) or an SPK1::GFP translational fusion (c, d) were obtained and GFP fluorescence was monitored. Scale bar = 100 µm.
(a)
(b)
(c) (d)
GFP
BUB1::GFP
SPK1::GFP