Present address: FAU Busan GmbH – Busan Branch, Busan Techno Park, 1276 Jisa-Dong, Gangseo-Gu, Busan 618-230, Republic of Korea.
AtHsp70-15-deficient Arabidopsis plants are characterized by reduced growth, a constitutive cytosolic protein response and enhanced resistance to TuMV
Article first published online: 28 APR 2011
© 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd
The Plant Journal
Volume 66, Issue 6, pages 983–995, June 2011
How to Cite
Jungkunz, I., Link, K., Vogel, F., Voll, L. M., Sonnewald, S. and Sonnewald, U. (2011), AtHsp70-15-deficient Arabidopsis plants are characterized by reduced growth, a constitutive cytosolic protein response and enhanced resistance to TuMV. The Plant Journal, 66: 983–995. doi: 10.1111/j.1365-313X.2011.04558.x
- Issue published online: 9 JUN 2011
- Article first published online: 28 APR 2011
- Accepted manuscript online: 21 MAR 2011 01:04PM EST
- Received 1 February 2011; accepted 22 February 2011; published online 28 April 2011.
Figure S1. Phenotype of hsp70-1/2 knockout (KO) plants and determination of TuMV titer. (a) Image of KO plants in comparison to wild type Col-0; (b) TuMV capsid protein was measured in younger systemic infected leaves 3 weeks after inoculation by an ELISA assay; the value of Col-0 wild type was set to 100% (with standard error bar, n = 20).
Figure S2. ABA treatment of adult amiRNA-AtHsp70-14/15 plants. Five days after repeated spraying with 0, 50 or 500 μM ABA, amiRNA-AtHsp70-14/15 and control plants were photographed.
Figure S3. Phenotype of TuMV infected amiRNA-Hsp70-14/15 plants. Five weeks old amiRNA-AtHsp70-14/15 and transformed control plants were inoculated with buffer or TuMV; approximately 4 weeks after inoculation amiRNA-AtHsp70-14/15 plants display less TuMV symptoms than control plants.
Figure S4. Transcriptional and phenotypic comparison of wild type, amiRNA-AtHsp70-14/15, hsp70-14 and hsp70-15 knockout (KO) plants. (a) The hierarchical cluster analysis of normalized microarray data from leaf RNA was performed with Genespring XI employing centered Pearson correlation with average linkage clustering algorithm. (b) Comparison of 6 weeks old plants before sampling for RNA extraction. The wild type-like phenotype of hsp70-14 KO and the phenotypic similarity of hsp70-15 KO and amiRNA-AtHsp70-14/15 plants is apparent.
Figure S5. Complementation of yeast sse1Δ mutant with AtHsp70-14/15. For the complementation experiment the growth deficient yeast strain sse1Δ (with W303 as corresponding wild type) was transformed with an expression vector for Hsp70-14/15-cmyc fusion proteins under control of a Galactose inducible promotor or a vector control. pENTR-AtHsp70-14/15 without stop codon (see Experimental Procedures) was recombinated by LR-CLONASE reaction (INVITROGEN, www.invitrogen.com) with the destination vector pYES-GW-3cmyc according to the manufacturer’s protocol to obtain the expression vector. For empty vector control pYES2 was used. (a) Serial dilutions of yeast strains (with optical dense at 600 nm of 1) were grown on YPD-Galactose medium at 37°C. (b) Expression of the Hsp70-14/15-cymyc fusion proteins was controlled by an immunoblot analysis with Anti-cmyc antibodies.
Table S1. Differentially regulated genes in amiRNA-Hsp70-14/15, hsp70-14 and hsp70-15 knockout plants. The list contains the differentially up-regulated (more than two-fold, P = 0.05) and down-regulated (<0.5-fold) genes between amiRNA-Hsp70-14/15, hsp70-14 and hsp70-15 knockout compared to wild type plants, as determined by microarray analysis. Values for absolute fold changes (FCAbsolute) are given.
Table S2. Primer combinations for RT-PCR analysis.
As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer-reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.
|TPJ_4558_sm_FigureS1.pdf||124K||Supporting info item|
|TPJ_4558_sm_FigureS2.pdf||136K||Supporting info item|
|TPJ_4558_sm_FigureS3.pdf||42K||Supporting info item|
|TPJ_4558_sm_FigureS4.pdf||525K||Supporting info item|
|TPJ_4558_sm_FigureS5.pdf||53K||Supporting info item|
|TPJ_4558_sm_Legends.doc||29K||Supporting info item|
|TPJ_4558_sm_TableS1.xls||234K||Supporting info item|
|TPJ_4558_sm_TableS2.doc||27K||Supporting info item|
|TPJ_4558_sm_TAIRannotationspreadsheet.xls||68K||Supporting info item|
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.