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Supplemental
Data.
de
Sena-Tomas
et
al.
(2011).
Plant
Cell
10.1105/tpc.110.082552
Supplemental Figure 1. Phosphorylation of Chk1 is required for normal in planta proliferation
of hyphae.
(A) Images of plant tissues 2 days post infection with compatible haploid strains carrying the
T394A S448A
non-phosphorytable chk1
allele. The picture shows hyphae (stained by WGA-AF488;
green) growing intracellularly in epidermal cells of maize (stained by propidium iodide; red).
T394A S448A
(B) Distribution of clamp-like structures in chk1
mutant cells. Infected plant tissue
(obtained after 2-3 days post-infection, dpi) was stained with WGA-AF488 and PI. Then
individual hyphae were counted and sorted in relation to the number of clamp-like structures
observed at the three more apical septa (S1, S2 and S3). Control (wild-type) filaments showed
the expected distribution of one clamp-like structure per septum. Mutant filaments showed a
clear deviation of this pattern. The graph shows the result from 2 independent experiments,
counting more than 30 filaments each.
Supplemental
Data.
de
Sena-Tomas
et
al.
(2011).
Plant
Cell
10.1105/tpc.110.082552
Supplemental Figure 2. Complementation of chk1 and atr1 deletion mutants
(A) Sensitivity of cells lacking chk1 or atr1 gene as well as their derivates with the
8
corresponding gene reintroduced at the ipp locus. A 10 cells/ml cell suspension and series of
10-fold dilutions were spotted (2 µl per spot) onto agar plates contained HU, hydroxyurea or UV
irradiated at the indicated dose after being spotted. The spots were photographed after
incubation for 2 days.
(B) Quantification of symptoms in maize plants after 14 days post-infection with wild-type,
chk1Δ, atr1Δ as well as complemented mutant crosses.
Supplemental
Data.
de
Sena-Tomas
et
al.
(2011).
Plant
Cell
10.1105/tpc.110.082552
Supplemental Figure 3. Single phosphorylation mutants in Chk1 showed wild-type phenotypes
with respect to cell cycle arrest and infection
T394A
(A) AB33 (control) and derived strains lacking the chk1 gene (chk1Δ) or carrying the chk1
S448A
and chk1
alleles were incubated in inducing conditions (MM-NO3). Cells were stained with
DAPI to detect nuclei. Graphic shows the percentage cells producing mononucleated filaments
(i. e., cell-cycle arrested) after 24 hours of incubation. The graph shows the result from two
independent experiments, counting more than 50 cells each. Means and standard deviations
are shown.
(B) Quantification of symptoms in maize plants after 14 days post-infection with wild-type,
T394A
S448A
chk1Δ, chk1
, and chk1
mutant crosses.
Supplemental
Data.
de
Sena-Tomas
et
al.
(2011).
Plant
Cell
10.1105/tpc.110.082552
Supplemental Figure 4. Absence of Brh2 or PolX does not affect the ability of b complex to
arrest cell cycle
(A) AB33 (control) and derived strains lacking the brh2 gene (brh2Δ) or the polX gene (polXΔ)
(were incubated in inducing conditions (MM-NO3). Cells were stained with DAPI to detect nuclei.
Graphic shows the percentage cells producing mononucleated filaments (i. e., cell-cycle
arrested) after 24 hours of incubation. The graph shows the result from two independent
experiments, counting more than 50 cells each. Means and standard deviations are shown.