Supplemental Figure 1： LecRK-V.2 and LecRK-VII.1 are necessary for the
resistance against to Pst.DC3000.
(A) The disease symptoms were observed 2-3 days after dip-inoculation with Pst
DC3000. (B) The bacterial growth were assessed 2-3 days after dip-inoculation with 5
x 106 cfu•ml-1 Pst DC3000 in Col-0 and lecrk mutants. Bacterial titers were evaluated
at 3d after inoculation (dpi). Values are the means ± SD of three biological replicates
(n = 9). Asterisks indicate a significant difference to Col-0 wild-type control based on
a t test (P <0.01). This experiments was repeated by Ph.D student, Shweta Bagul and
the other former members in the Zimmerli laboratory.
Supplemental Figure 2： LecRK-V.2 and LecRK-VII.1 are necessary for stomatal
immunity.
(A and B) The stomata aperture from the epidermal peels of Col-0, lecrk-V.2, and
lecrk-VII.1 mutant lines. Epidermal peels were exposed to MES buffer (Mock) or 5 x
106 cfu•mL-1 Pst DC3000 for 1.5 or 3 hours. (C) The stomatal aperture from the
epidermal peels of Col-0, lecrk-V.2, and lecrk-VII.1 mutant lines after 3 hours of
incubation in flg22 with 0 (Mock), 0.5, 1 or 5 μM, respectively.
Supplemental Figure 3：The PTI apoplastic responses and regulations in lecrk-V.2
and lecrk-VII.1 mutant lines.
(A and B) The reactive oxygen species productions in Arabidopsis leaves of Col-0,
lecrk-V.2 and lecrk-VII.1 mutant lines after treatment with 1 μM flg22 as relative light
units (RLU). The bak1-4 mutant line is used as negative control. Values are the means
± SE (n = 6). (C) Callose deposition in Col-0, lecrk-V.2 and lecrk-VII.1 mutant leaves
9 hours after infiltration with 10 mM MgSO4 (Mock) or 1 x 108 cfu•mL-1 Pst DC3000
hrcC. Data are means ± SD of callose deposits per square millimeter (n = 9).
Differences among Col-0, lecrk-V.2 and lecrk-VII.1 mutant were not significantly
different base on a t test (P < 0.01). Bar = 400μm. (D) FRK1 expression levels in
10 mM MgSO4 (Mock) or 1 x 108 cfu•mL-1 Pst DC3000 hrcC (h). Transcripts levels
determined by qRT-PCR and normalized to both EF-1 and UBQ10 were compared to
Col-0 mock at 45 minutes (defined value of 1). Three independent biological replicated
performed in triplicate were average ± SD (n = 9). Differences among Col-0,
lecrk-V.2 and lecrk-VII.1 mutant were not significantly different base on a t test (P <
0.01).
Supplemental Figure 4：LecRK-V.2-GFP and LecRK-VII.1 –GFP fusion proteins
localized on the membrane in Arabidopsis mesophyll protoplast.
LecRK-V.2-GFP and LecRK-VII.1–GFP fusion proteins were driven by the
cauliflower mosaic virus 35S promoter and transiently expressed in Arabidopsis
mesophyll protoplast. (A and D) GFP fluorescence channel. (B and E) Plasma
membrane marker pm-rk CD3-1007-mCherry fluorescence localization. (C and F)
Overlapping images of the GFP, plasma membrane marker and chlorophyll (red)
autofluorescence. (G) Protoplast expressing the GFP vector only as a GFP control.
Supplemental Figure 5：LecRK-V.2 and LecRK-VII.1 interact with FLS2 only
after the treatment with flg22.
In vivo, the interactions between FLS2 and LecRK-V.2 or FLS2 and LecRK-VII.1 are
determined by performing bimolecular fluorescence complementation (BiFC) analyses.
LecRK-V.2-GFP and LecRK-VII.1–GFP fusion proteins were under the control of the
cauliflower mosaic virus (CaMV) 35S promoter. FLS2-YFPN and BAK1-YFPC,
FLS2-YFPN and LecRK-V.2-YFPC, FLS2-YFPN and LecRK-VII.1-YFPC were
transiently co-transformed into Arabidopsis protoplast and with (+) or without (-) the
treatment of 1μM flg22 for 10 minutes. The red and yellow signals are
autofluorescence and YFP, respectively. There were the image of bright field and the
merge which is combined the autofluorescence, YFP, and bright field image.
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