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Figure 1.

Differential phenotypes of L. japonicus ecotypes MG-20 and Gifu B-129 against Pto.

A. Symptom development following bacterial infiltration in leaves from both ecotypes. Central leaflets were infiltrated using a needleless syringe as described in Materials and Methods. Eventually, lateral leaflets were marked with a black pen just for identification purposes. Pictures were taken immediately before inoculation and after a period of 24, 48 or 72 h. B. Measurement of Pto growth within MG-20 and Gifu B-129 infiltrated leaves. Data points represent the mean ± SD of three replicate experiments. Bacterial levels at 24, 48 and 72 hpi were statistical analyzed and compared against bacterial levels at 0 hpi, ** p<0.01. Black squares, MG-20; white squares, Gifu B-129.

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Figure 2.

Effect of bacterial infiltration on ROS accumulation in L. japonicus leaves.

Detached leaves of each ecotype were submerged for 30 min in a solution of 15 µM DCFDA and infiltrated with a bacterial solution or MgCl2 (controls). Then, leaves were incubated in freshly prepared DCFDA solution and observed under fluorescence microscopy at indicated time points (excitation filter, 460 nm; emission filter, >515 nm). Figure shows representative photographs of control and treated leaves. Bars: 500 µm.

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Figure 3.

MapMan illustration depicting the transcripts from the “Biotic stress” BIN regulated under pathogenic conditions.

Transcriptomic data from bacteria-inoculated leaves were compared to their respective controls (mock-inoculated). Genes that were shown to be differentially expressed were mapped using the MapMan software (http://mapman.gabipd.org). Picture shows genes related to biotic stress regulated in MG-20 (A) and Gifu (B). Log fold change ratios are indicated as a gradient of red (down-regulated) and green (up-regulated).

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Figure 4.

Trend plots comparing quantitative expression of differentially regulated genes in MG-20 and Gifu B-129.

Gene order along the x axis was determined by the expression level in Gifu B-129 samples (dark lines). The expression of the same genes in MG-20 (grey lines) was plotted keeping the same order.

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Figure 5.

Comparison of microarray and quantitative real-time PCR data for selected genes.

Symbols represent mean expression levels in bacterial-infiltrated relative to mock-inoculated leaflets of Gifu B-129 (●) or MG-20 (○).

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Figure 6.

MapMan illustration depicting biotic stress-related transcripts with a relative higher expression when mock-inoculated samples are compared.

Transcriptomic data from mock-inoculated leaves of MG-20 and Gifu B-129 were compared to each other. Genes that were shown to be differentially expressed in MG-20 (A) and Gifu B-129 (B) were mapped using the MapMan software. Picture shows only those genes related to biotic stress. Log fold change ratios are indicated as a gradient of green.

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