1. Volume 5, Issue 5, Pages 1113-1124 (September 2012)
The LysM Receptor-Like Kinase LysM RLK1 Is Required to Activate Defense and Abiotic-Stress Responses Induced by Overexpression of Fungal Chitinases in Arabidopsis Plants 
Yariv Brotman, Udi Landau, Smadar Pnini, Jan Lisec, Salma Balazadeh, Bernd Mueller-Roeber, Aviah Zilberstein, Lothar Willmitzer, Ilan Chet, Ada Viterbo 
Molecular Plant 
Volume 5, Issue 5, Pages (September 2012)
DOI: /mp/sss021
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

2. Figure 1
Detection of Fungal Chitinase Expression in Arabidopsis Lines and Tolerance to Salt Stress.
(A) T-DNA region of the binary vector used for transforming Arabidopsis. FL, fungal leader peptide sequence; HA, intron-disrupted HA epitope.
(B) Western blot analysis of total leaf proteins of selected Arabidopsis homozygous transgenic lines harboring the ectopic chitinase genes or the T-DNA of the empty vector control (EV).
(C) Salt-stress tolerance. Arabidopsis seeds were germinated on B5 medium supplemented with 100 or 150 mM NaCl. Results are the average of three independent experiments. In each experiment, four plates were analyzed per line, with 30–40 seeds per plate. Germinating seeds were counted after 4 d (untreated control plants show 100% germination rate).
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

3. Figure 2 Tolerance to Heavy Metals.
Arabidopsis seeds were germinated on B5 medium with (A) 200 μM CdSO4 or (B) 150 or 300 μM CuSO4. Germinating seeds were counted after 4 d (untreated control plants show 100% germination rate). Seedlings with green cotyledons were counted after 5 and 10 d. Results are averages of three independent experiments. In each experiment, four plates were analyzed for each line, with 30–40 seeds per plate. Means ± SE are indicated. *, the line marked with an asterisk is significantly different from all the other lines in the same panel (P < 0.05).
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

4. Figure 3 Tolerance to B. cinerea Infection.
(A) Detached leaves of EV and chitinase-overexpressing lines 2.17, 3.5, 5.9, and 7.6 were infected with a spore suspension as described in the ‘Methods’ section. Disease symptoms were recorded after 3 d and classified according to a disease index scale from 0 (no symptom) to 5 (large necrotic area).
(B) Tolerance to B. cinerea infection was evaluated in the lysM 2.17 hybrid line, the parental lines LysM RLK1 and 2.17, and in EV. A total of 64 leaves were infected for each line in four independent experiments. *, line 2.17 marked with asterisk is significantly different from all the other lines in both panels (P < 0.05).
Molecular Plant 2012 5, DOI: ( /mp/sss021)
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5. Figure 4
Chitin-Eliciting Effect on Chlorophyll Content in Salt-Treated EV and LysM RLK1 Mutant Line.
Total chlorophyll (Chl) concentration (mg g−1 leaves) was measured in EV and LysM RLK1 mutant seedlings after 4 d of growth in the absence or presence of 150 mM NaCl pretreated (24 h) with chitin (100 μg ml−1) or 1 μM of the chitin pentamer. C, untreated. Comparisons between groups are indicted with bars. *, significant difference (P < 0.05).
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

6. Figure 5
Tolerance to Salt and Heavy Metals in Chitinase-Overexpressing Lines Crossed to the LysM RLK1 Mutant.
Arabidopsis seeds of EV, lysm rlk1 mutant, chitinase-overexpressing lines 2.17, 3.5, 5.9, and hybrid lines lysm2.17, lysm 3.5, and lysm 5.9 were germinated on B5 medium with 125 mM NaCl (A), 150 μM CuSO4 (B), or 200 μM CdSO4 (C). Germinating seeds were counted after 4 d (A, B). Seedlings with green cotyledons were counted after 10 d. Means ± SE are indicated. *, Lines marked with asterisk are significantly different from all the other lines in three panels (P < 0.05).
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

7. Figure 6
Gene Expression (Relative to EV) in the Chitinase-Overexpressing Line 2.17, LysM RLK1 Mutant, and lysM 2.17 Line.Venn diagrams showing the overlap of the up-regulated and down-regulated genes (at least twofold) in the chitinase-overexpressing line 2.17, LysM RLK1 mutant, and lysM 2.17 line.
(A) Chitin-responsive and other defense-related genes.
(B) ROS-responsive genes. Numbers in brackets indicate the total number of genes up- or down-regulated in each line. The list of genes that show significant difference (P < 0.05) is displayed in Supplemental Table 2.
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

8. Figure 7
Heat Map Representation of Genes Differentially Up-Regulated in the 2.17 Chitinase-Overexpressing Line during Salt Stress.
Fold expression (more than twofold) relative to EV was measured 1 and 3 h after the addition of 150 mM NaCl to the growth medium. The list of genes that show a significant difference (P < 0.05) to EV is provided in Supplemental Table 3.
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions

9. Figure 8
Effect of NaCl on the Expression of MYB51, BGLU27, CYP81F2, STZ, PAD3, and WRKY30 Genes in 2.17, LysM RLK1 Mutant, and lysM 2.17.mRNA levels were measured in Arabidopsis seedlings before stress treatment and 1 and 3 h after the treatment with 150 mM NaCl. Relative expression levels, measured by qRT–PCR, are shown as fold change values relative to EV. Gray columns, 2.17; white columns, LysM RLK1 mutant; black columns, lysM 2.17.
Molecular Plant 2012 5, DOI: ( /mp/sss021)
Copyright © 2012 The Authors. All rights reserved. Terms and Conditions