DEV02340 Supplementary Material

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• Supplemental Tables - Adobe PDF

• Supplemental Figure 1 -

  Fig. S1. Characterization of msi1 mutants, MSI1 antisense and msi1-tap1 plants. (A) MSI1 antisense plants contain reduced MSI1 protein amounts. Protein was extracted from leaves of wild-type control plants (WT) and the transgenic line MSI1AS7b (msi1-as) expressing a fragment of the MSI1 cDNA in antisense orientation. Ten μg protein per sample was subjected to immunoblotting with anti-MSI1 antiserum. (B) Analysis of the flowering time of MSI1/msi1 plants grown in short and long days (SD and LD, respectively). (C) Analysis of the flowering time of F1 WT × msi1-tap1 and msi1-tap1 × WT plants grown in long days (LD). (D) MSI1/msi1 plants contain normal MSI1 protein amounts. Protein was extracted from leaves of wild-type and msi1/MSI1 plants. Ten μg protein per sample was subjected to immunoblotting with anti-MSI1 antiserum. (E) 35-day-old wild-type (WT), msi1-tap1 and msi1-tap1 35S::MSI1 plants grown in LD. Note the absence of phenotypic traits of MSI1 co-suppression plants, such as arrest of the primary shoot or sterility in the analyzed msi1-tap1 35S::MSI1 plants. (F) Dose-response curves for the promotion of flowering by gibberellic acid (GA₃) in wild-type and msi1-tap1 plants in LD. 100% represents the flowering time (number of rosette leaves at bolting) of untreated plants.

• Supplemental Figure 2 -

  Fig. S2. Characterization of insertion mutants used in our study. (A) Absence of CLF transcripts from the used clf allele. (B) Expression of MEA in clf and msi1-tap1. For A and B, RNA was extracted from rosette leaves of 34-day-old plants grown on soil in LD. (C) Absence of FLC and MSI4/FVE transcripts from the used flc and fve alleles, respectively. (D) Absence of FLM transcripts from the used flm allele. FLM transcripts are as described previously (Scortecci et al., 2001). For C and D, RNA was extracted from 8-day-old seedlings grown on plates containing MS medium in LD. (E) Expression of FT in msi1-tap1 seedlings and in wild-type inflorescences. RNA was extracted from 8-day-old seedlings grown on plates containing MS medium in LD and from inflorescences (infl.) of 35-day-old plants grown on soil in LD. Note the strong signal in wild-type inflorescences, demonstrating that the PCR was not saturated. n.t., no template control. For all experiments, GAPDH was used as a control.

  Scortecci, K. C., Michaels, S. D. and Amasino, R. M. (2001). Identification of a MADS-box gene, FLOWERING LOCUS M, that represses flowering. Plant J. 26, 229-236.

• Supplemental Figure 3 -

  Fig. S3. Distribution of pair-wise Pearson correlation coefficients of gene expression signals. Data were taken from the AtGenExpress developmental series consisting of 238 microarrays (Schmid et al., 2005). Affymetrix *.CEL files were analyzed using the GCRMA algorithm. The estimate of the correlation coefficient distribution is based on 100,000 randomly selected pairs of genes. Note that most genes have little pair-wise correlation of expression signals during development. There are slightly more gene-pairs with a high positive (close to 1) than with a high negative correlation (close to -1). MSI1 and MSI4/FVE have a Pearson correlation coefficient of 0.92, which is among the top 0.028% of all values.

  Schmid, M., Davison, T. S., Henz, S. R., Pape, U. J., Demar, M., Vingron, M., Scholkopf, B., Weigel, D. and Lohmann, J. U. (2005). A gene expression map of Arabidopsis thaliana development. Nat. Genet. 37, 501-506.

• Supplemental Figure 4 -

  Fig. S4. MSI1 is strongly expressed in the shoot apex. Data are from transcriptional profiling of various tissues (Schmid et al., 2005) and from transcriptional profiling of apices from plants grown in SD after transfer into LD to induce flowering (Schmid et al., 2003). Affymetrix ATH1 microarrays were used in both studies. Raw data were re-analysed using the MAS5 algorithm implemented in Bioconductor.

  Gentleman, R. C., Carey, V. J., Bates, D. M., Bolstad, B., Dettling, M., Dudoit, S., Ellis, B., Gautier. L., Ge, Y., Gentry, J. et al. (2004). Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80.

  Schmid, M., Uhlenhaut, N. H., Godard, F., Demar, M., Bressan, R., Weigel, D. and Lohmann, J. U. (2003). Dissection of floral induction pathways using global expression analysis. Development 130, 6001-6012.

  Schmid, M., Davison, T. S., Henz, S. R., Pape, U. J., Demar, M., Vingron, M., Scholkopf, B., Weigel, D. and Lohmann, J. U. (2005). A gene expression map of Arabidopsis thaliana development. Nat. Genet. 37, 501-506.

• Supplemental Figure 5 -

  Fig. S5. Alignment of MSI1 and MSI4 amino acid sequences. The alignment was generated using GeneDoc (see http://www.psc.edu/biomed/genedoc/)