Dual involvement of a Medicago truncatula NAC transcription factor in root abiotic stress response and symbiotic nodule senescence
Article first published online: 10 JAN 2012
© 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd
The Plant Journal
Volume 70, Issue 2, pages 220–230, April 2012
How to Cite
de Zélicourt, A., Diet, A., Marion, J., Laffont, C., Ariel, F., Moison, M., Zahaf, O., Crespi, M., Gruber, V. and Frugier, F. (2012), Dual involvement of a Medicago truncatula NAC transcription factor in root abiotic stress response and symbiotic nodule senescence. The Plant Journal, 70: 220–230. doi: 10.1111/j.1365-313X.2011.04859.x
- Issue published online: 3 APR 2012
- Article first published online: 10 JAN 2012
- Accepted manuscript online: 18 NOV 2011 11:43PM EST
- Received 24 October 2011; accepted 15 November 2011; published online 10 January 2012.
Figure S1. Expression analysis of MtNAC969 and related M. truncatula NAC genes in composite plants overexpressing or downregulating MtNAC969.A. Expression analysis of MtNAC969 and its 2 closest homologs, TC80936 and TC148340 (MtGI v8), in roots of GUS RNAi and MtNAC969 RNAi composite plants under control (left graph) or salt stress (right graph) conditions. B. MtNAC969 expression in 35S::GUS and 35S::MtNAC969 composite plants in control (left graph) and salt stress (100 mM NaCl, 1 week; right graph) conditions. Bar graphs represent the quantification of specific PCR amplification products for MtNAC969, TC80936 and TC148340, normalized using the average of 3 constitutive genes (selected as described in methods). The GUS RNAi Plant1 was used as calibrator to define fold changes (indicated by the broken line). For each condition and genotype, 2 plants are shown and error bars represent confidence interval (α=0.05) of 2 technical replicates.
Figure S2. Phylogenetic tree of selected NAC proteins and MtNAC969 / AtNAP sequence comparison. A. The phylogenetic tree includes the three salt-regulated NACs previously identified (Gruber et al., 2009) as well as the M. truncatula NAC protein most closely related to MtNAC969 (TC148340, MtGI version 8 database). Arabidopsis and rice NAC proteins, selected based on their functional characterization regarding abiotic stresses, are also included. Alignments of full length amino acid sequences were done using ClustalW, and the tree was constructed with the MEGA 4 software (see Methods for details).
B. Alignment of amino acid sequences of MtNAC969 and AtNAP using the CLUSTALW software. Identical amino acids are highlighted in grey, and the NAC domain of MtNAC969 predicted using PROSITE (http://www.expasy.ch/prosite/) is underlined.
Figure S3. MtNAC969 gene expression profiles based on the Medicago truncatula Gene Expression Atlas (MtGEA). A. MtNAC969 expression in different organs. B. MtNAC969 expression in roots in response to 1 hour abiotic stresses: 100 mM NaCl, 37°C, 4°C and 200 mM mannitol. Bar graphs represent the quantification of specific PCR amplification products for MtNAC969, normalized using the average of 3 constitutive genes (selected as described in methods). Fold changes are indicated by the broken line relatively to the non-treated condition. A representative example out of 2 biological experiment is shown, and errors bars represent confidence interval of 2 technical replicates (α=0.05). C. MtNAC969 root expression in response to biotic, abiotic and symbiotic conditions. In A and C, values correspond to mean transcript levels obtained using Affymetrix chips in the specified biological conditions, as described in Benedito et al. (2008) and available at http://mtgea.noble.org/v2/ .
Table S1. List of primers used. In italics is indicated the T7 promoter extension used to synthesize the RNA probes for in situ hybridization. IDs correspond to MtGI version 8 or NCBI databases.
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