SEARCH

SEARCH BY CITATION

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.

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.

FilenameFormatSizeDescription
TPJ_4859_sm_FigS1-S3.pdf92KSupporting info item
TPJ_4859_sm_LegendsforSupportingInformation.dot28KSupporting info item
TPJ_4859_sm_TableS1.doc56KSupporting 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.