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. 2017 Jun 15;7:3557. doi: 10.1038/s41598-017-03704-x

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Pseudomonas syringae pv. actinidiae type III effector HopZ5 triggers accession-specific immunity in Arabidopsis. (a) Arabidopsis accession-dependent development of a hypersensitive response to P. syringae pv. actinidiae and its type-III secreted effector HopZ5. Leaves of Arabidopsis plants were infiltrated with P. syringae pv. actinidiae NZV-13 or P. fluorescens Pf0-1 (T3S) carrying an empty vector pBBR1MCS-5 (EV), hopZ5 with a C-terminal 6xHA tag under the control of the avrRps4 promoter in pBBR1MCS-5 or untagged avrRpm1 under its native promoter in pVSP61. Bacterial suspensions (1 × 10⁸ CFU/mL) were blunt-syringe infiltrated into the leaves and photographs were taken 20 hours after infiltration (1 dpi). Development of the hypersensitive response is indicated by a red asterisk. (b) Electrolyte leakage from Arabidopsis leaf discs after infiltration with P. fluorescens Pf0-1 (T3S) carrying the indicated constructs, as in (a). Bacteria (1 × 10⁸ CFU/mL) were blunt syringe-infiltrated into the leaves. The error bars indicate the standard error from four technical replicates. The experiment was conducted three times with similar results. (c) P. syringae pv. tomato DC3000 (Pto DC3000) growth in Arabidopsis. Pto DC3000 carrying EV, hopZ5-HA or avrRpm1 was blunt syringe-infiltrated at 5 × 10⁵ CFU/mL into Arabidopsis leaves, and bacterial growth was determined 4 days post-infection (4 dpi). Error bars represent standard error from six technical replicates. Asterisks indicate results of Student’s t-test between selected sample and EV for that accession; *(P < 0.05), **(P < 0.01). The experiment was conducted six times with similar results. (d) Defence gene expression in Arabidopsis in response to EV, HopZ5-HA or AvrRpm1. P. fluorescens Pf0-1 (T3S) (1 × 10⁸ CFU/mL) carrying EV, hopZ5-HA or avrRpm1 was blunt syringe-infiltrated into leaves, samples taken at 8 hours post infiltration and defence gene expression determined from extracted RNA by quantitative polymerase chain reaction. Expression for each defence gene is relative to internal EF1α expression and defence gene expression for EV samples indicated. Error bars indicate standard error from three technical replicates. Asterisks indicate results of Student’s t-test between selected sample and EV for that accession; *(P < 0.01). The experiment was conducted three times with similar results.