File S1 - Alterations in Auxin Homeostasis Suppress Defects in Cell Wall Function

Supplemental material. Figure S1, T-DNA insertion in IAR4 results in reduced transcript levels. (A) Cartoon of the IAR4 gene: boxes represent exons, line introns. The site of the T-DNA insertion in iar4-7 is indicated by the red triangle and primers used for the analysis of transcript levels indicated below. (B) IAR4 transcript levels of different regions (as indicated in (A)); n = 3 (± SE). Transcript levels were calculated using the 2(−ΔΔCt) method as described [51]. Asterisks indicate significant differences between iar4-7 and the wild type or fei1 fei2 iar4-7 and fei1 fei2 (P<0.05); nd, not detectable. Figure S2, Complementation test of iar4 alleles. F1 progeny from indicated crossed were grown on MS media containing 4.5% sucrose for 14 days. Note that the roots of the F1 seedlings display a non-swollen (i.e. suppressed) phenotype, suggesting that the iar4-5, iar4-6 and iar4-7 mutations are allelic. Scale bar = 0.5 mm. Figure S3, Effect of altered auxin levels on fei1 fei2 root growth and on the suppression of fei1 fei2 by iar4. (A) Effect of elevated temperature on the suppression of root swelling of the fei1 fei2 mutant by iar4-5. Four-day old seedlings grown on MS with no sucrose at 22°C were transferred to MS media containing 4.5% sucrose and grown an additional five days at 28°C. (B) Quantification of root elongation of wild-type and fei1fei2 in response to auxin. Four-day-old seedlings were transferred to media containing the indicated level of auxin and the amount the roots grew after transfer was measured four days later. Values represent the mean of ± SE (n>15). (C) Quantification of relative root elongation of wild-type and fei1fei2 as in described in A. Values were normalized to the no auxin control. Data were analyzed by Student's t-test; *, P<0.05. **, P<0.01. The experiment was repeated three times and showed very similar results. Figure S4, iar4-5 restores fertility of fei1 fei2 cob-1. (A) Inflorescence, (B) flower and (C) silique phenotypes of indicated genotypes. Note that iar4-5 partially restores the silique length of fei1 fei2 cob. Plants were grown on soil under long day conditions for four weeks. To visualize stamen length, some petals and sepals were removed from each flower. Bar = 1 mm. (D) Carpel length of indicated genotypes. Different letters indicate significant differences between groups. Data were tested with one-way ANOVA and Tukey's post-hoc analysis; n>13; P<0.05. Table S1, Markers used to map iar4-5. Table S2, Primers used for gene expression analysis.
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