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Open Access Research article

Loss of compatibility might explain resistance of the Arabidopsis thaliana accession Te-0 to Golovinomyces cichoracearum

Georgina Fabro and María Elena Alvarez*

Author Affiliations

Centro de Investigaciones en Química Biológica de Córdoba CIQUIBIC, UNC-CONICET, Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Haya de la Torre y Medina Allende, Ciudad Universitaria, Córdoba, X5000HUA, Argentina

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BMC Plant Biology 2012, 12:143  doi:10.1186/1471-2229-12-143

Published: 11 August 2012

Additional files

Additional file 1:

a) Time-course quantification of cellular defence responses to G. cichoracearum in Te-0, Col-0 and Kas-1 plants. 1- DAB stained cells in contact with G. cichoracearum structures. (*) No significant differences between these values in T-test at p < 0.05. 2- Trypan blue stained plant cells observed by bright field microscopy. 3- Haustoria stained with SRB-ANS were observed by confocal microscopy at the stated times post-inoculation. Abnormal haustoria are shown in Figure 5 (Kas-1, 48 hpi). In all these experiments 150 fungal interaction sites (1,2) or 150 haustoria (3) were evaluated for each genotype at the indicated times. Three independent experiments were performed. Values represent weighted average percentage ± coeficent of variance of the independent experiments. b) Mesophyll cell death evaluated on Kas-1 leaves infected with G. cichoracearum at 48 hpi by trypan blue staining. Left: focus on epidermal cells interacting with the pathogen. Right: underlying mesophyll dead cells.

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Additional file 2:

Expression of defence genes on G. cichoracearum-infected tissues assessed by qRT-PCR. Values were obtained applying the ΔΔCt method using UBQ5 as housekeeping gene control. Bars represent average ± SD from three replicates.

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Additional file 3:

Time course development of G. cichoracearum infective structures on Col-0, Te-0 and Kas-1 accessions. The abundance of different fungal structures is evaluated at the indicated time points. PGT: primary germ tube, HA: haustorium, SGT: secondary germ tube, R: ramified hyphae colony. Tissues were inoculated with the same conidial suspension. Values are referred as percentages of inoculated conidia.

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Additional file 4:

DAPI staining of the G. cichoracearum haustoria developed in Col-0, Te-0 and Kas-1 plants. Haustoria were co-stained with SRB, ANS and DAPI. Z-stacks are shown for representative haustoria at 72 hpi. HN: haustorial nucleus. Bars: 10 μm.

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Additional file 5:

Uptake of ArgC14 by G. cichoracearum haustoria installed in Te-0 cells. a) Uptake of ArgC14 by the fungal tissues differentiated on Col-0, Te-0 and Kas-1 leaves. The indicated values were obtained after subtracting residual radioactivity levels present on tapes stripped from uninfected leaves (see experimental procedures). b) ArgC14 levels detected in fungal tissue expressed as a ratio over the number of haustoria present on sibling leaf discs. c) Time-course estimation of the fungal biomass found on the surface of Col-0 and Te-0 leaves infected with G. cichoracearum. Fifteen leaves per plant were analyzed per time-point. Average percentage ± SD of leaf areas covered by fungal colonies is depicted.

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Additional file 6:

Genes with differential expression in susceptible Col-0 and Te-0 resistant plants during the interaction with G. cichoracearum evaluated by q-RT-PCR. Fold changes in transcript expression levels were determined as described in Additional file 2. Bars represent average ± SD from three replicates, except for STP4, where q-PCR assay included two replicates.

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Additional file 7:

Uninfected Te-0 plants do not accumulate callose. Untreated leaves from Te-0 plants (5-week-old, soil grown) were collected and stained with aniline blue to reveal callose by confocal microscopy using UV light. This assay was performed in parallel, under identical conditions to that shown in Figure 1b, where deposition of callose was detected in Kas-1 infected samples. Col-0 was included for comparison.

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Additional file 8:

Primers and conditions used for semi-quantitative and real time/quantitative RT-PCR assays. (Ψ) Number of cycles used for sq-RT-PCR. (*) Different melting temperatures (Tm) were used in sq and q-RT-PCR. All q-RT-PCR were done with Tm = 60°C, except for PDF1-2. (^) UBQ5 and GapC were used as housekeeping control genes in q-RT-PCR and sq-RT-PCR, respectively.

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