The Arabidopsis thaliana transcriptional activator STYLISH1 regulates genes affecting stamen development, cell expansion and timing of flowering
Plant Molecular Biology
Veronika Ståldal1, Izabela Cierlik1, Song Chen1, Katarina Landberg1, Tammy Baylis2, Mattias Myrenås1, Jens F Sundström1, D Magnus Eklund1, Karin Ljung3 and Eva Sundberg1
Corresponding author:
Department of Plant Biology and Forest Genetics, Uppsala BioCenter, Swedish University of Agricultural Sciences, PO-Box 7080, SE-750 07 Uppsala, Sweden
E-mail:
Tel: +46 18 673245
Fax: +46 18 673389
Supplementary table I. Microarray data of selected genes
AGI code / Array, 35Spro:STY1:GR sty1-1 sty2-1 DEX/MockAffymetrix / CATMA
1h / 3h / 2h / 3h / 6h
At5g11320 / 21.6 / 3.3 / 4.8 / 5.0
At1g56710 / 18.2 / 4.8 / 10.4 / 11.4
At4g31610
At1g06160 / 2.3
At2g36630 / 2.4 / 2.8a / 2.5
At1g75430 / 3.0 / 5.6
At2g31230
At4g32800 / 3.3
At3g51660 / 2.2a / 2.2
At2g46660
At3g61970 / 2.3
At5g21105 / 2.6 / 3.0
At4g30250b / 2.9 / 3.0
At1g21670 / 2.2
At4g38400 / 4.5 / 3.8 / 2.5 / 2.9
At1g56020 / 2.1a
At4g29720 / 2.3 / 2.5 / 3.0 / 3.1 / 2.5
At4g28720
At2g35700 / 3.2
Levels of DEX induced upregulation of the selected genes in 35Spro:STY1:GR sty1-1 sty2-1 seedlings compared to mock treated seedlings detected in microarray experiments at different time points after DEX treatment
Values represent the mean of two (Affymetrix array) or three (CATMA array) biological replicates
a indicates that the value represents one biological replicate
b indicates that this gene is not present on Affymetrix ATH1 array
Supplementary table II. Co-expression of selected genes with SHI/STY genes
MR / Cor / MR / Cor / MR / Cor / MR / Cor / MR / Cor / MR / Cor
At5g11320 / 36 / 0.45 / 285 / 0.29
At1g56710
At4g31610 / 239 / 0.30 / 277 / 0.37 / 79 / 0.44 / 104 / 0.42 / 190 / 0.48
At1g06160
At2g36630
At1g75430 / 162 / 0.32 / 26 / 0.46 / 103 / 0.49
At2g31230
At4g32800
At3g51660
At2g46660
At3g61970
At5g21105
At4g30250a
At1g21670
At4g38400
At1g56020 / 70 / 0.41 / 201 / 0.37 / 36 / 0.52
At4g29720
At4g28720
At2g35700
Co-expression with SHI/STY-family genes and YUC4 in various published microarray experiments (STY1, SRS3 and SRS7 are not present on Affymetrix ATH1 array and high co-expression was not found with SRS6)
MR indicates mutual rank
Cor indicates correlation
Only values for a mutual rank of 300 or lower are shown
a indicates that this gene is not present on Affymetrix ATH1 array
Supplementary table III. qRT-PCR detected expression of selected genes in 35Spro:STY1:GR sty1-1 sty2-1 seedlings
Mock / DEX / CHX / CHX+DEX
At5g11320 / 1 ± 0.1 / 31 ± 10 / 2.6 ± 0.8 / 66 ± 16
At1g56710 / 1 ± 0.9 / 4.5 ± 0.2 / 0.18 ± 0.002 / 3.7 ± 0.6
At4g31610 / 1 ± 0.2 / 13 ± 2 / 2.8 ± 0.1 / 5.4 ± 0.9
At1g06160 / 1 ± 0.2 / 1.1 ± 0.3 / 1.2 ± 0.3 / 8.1 ± 1
At2g36630 / 1 ± 0.2 / 3.0 ± 0.5 / 1.7 ± 0.1 / 2.7 ± 0.2
At1g75430 / 1 ± 0.1 / 7.2 ± 1 / 4.2 ± 0.6 / 74 ± 1
At2g31230 / 1 ± 0.3 / 1.0 ± 0.1 / 1.4 ± 0.3 / 5.4 ± 0.4
At4g32800 / 1 ± 0.5 / 1.1 ± 0.1 / 7.8 ± 2 / 18 ± 2
At3g51660 / 1 ± 0.1 / 1.4 ± 0.04 / 1.7 ± 0.3 / 2.8 ± 0.3
At2g46660 / 1 ± 0.3 / 4.3 ± 0.9 / 1.2 ± 0.09 / 4.3 ± 0.8
At3g61970 / 1 ± 0.08 / 5.2 ± 0.8 / 1.7 ± 0.06 / 9.1 ± 0.6
At5g21105 / 1 ± 0.2 / 1.6 ± 0.1 / 1.4 ± 0.1 / 2.4 ± 0.2
At4g30250 / 1 ± 0.3 / 7.2 ± 0.5 / 2.7 ± 0.5 / 7.7 ± 0.8
At1g21670 / 1 ± 0.3 / 2.8 ± 0.6 / 1.0 ± 0.2 / 2.3 ± 0.1
At4g38400 / 1 ± 0.3 / 3.8 ± 0.7 / 4.8 ± 0.6 / 14 ± 3
At1g56020 / 1 ± 0.1 / 1.8 ± 0.3 / 3.4 ± 0.4 / 7.2 ± 0.6
At4g29720 / 1 ± 0.07 / 2.0 ± 0.2 / 1.6 ± 0.05 / 6.5 ± 0.6
At4g28720 / 1 ± 0.2 / 1.2 ± 0.3 / 2.0 ± 0.4 / 4.0 ± 0.3
At2g35700 / 1 ± 0.2 / 3.2 ± 0.08 / 1.0 ± 0.01 / 4.4 ± 0.2
Values are relative expression values representing the mean of three biological replicates ± standard error of the mean.
Supplementary table IV. qRT-PCR detected expression of selected genes in SHI/STY-family quintuple mutant and wild type plants
Quintuple / Col / Ler / Quintuple / Col / Ler
At5g11320 / 10 ± 6 / 15 ± 6 / 1 ± 0.4 / 1.4 ± 0.4 / 2.5 ± 0.6 / 1 ± 0.3
At1g56710 / 0.1 ± 0.09 / 0.9 ± 0.2 / 1 ± 0.2 / 0.3 ± 0.07 / 1.2 ± 0.1 / 1 ± 0.1
At4g31610 / 0.2 ± 0.1 / 1.1 ± 0.1 / 1 ± 0.09 / 0.4 ± 0.04 / 1.1 ± 0.04 / 1 ± 0.1
At1g06160 / 0.4 ± 0.06 / 0.7 ± 0.1 / 1 ± 0.1 / 0.5 ± 0.01 / 4.0 ± 0.8 / 1 ± 0.1
At2g36630 / 0.5 ± 0.02 / 0.7 ± 0.03 / 1 ± 0.1 / 0.7 ± 0.09 / 1.7 ± 0.1 / 1 ± 0.1
At1g75430 / 0.5 ± 0.06 / 1.6 ± 0.2 / 1 ± 0.05 / 1.0 ± 0.1 / 1.7 ± 0.2 / 1 ± 0.1
At2g31230 / 1.4 ± 0.1 / 1.0 ± 0.1 / 1 ± 0.1 / 0.4 ± 0.03 / 1.4 ± 0.2 / 1 ± 0.2
At4g32800 / 1.4 ± 0.2 / 1.2 ± 0.2 / 1 ± 0.06 / 0.6 ± 0.05 / 1.1 ± 0.06 / 1 ± 0.08
At3g51660 / 1.4 ± 0.2 / 1.6 ± 0.2 / 1 ± 0.1 / 0.8 ± 0.07 / 1.9 ± 0.3 / 1 ± 0.2
At2g46660 / 0.7 ± 0.09 / 0.9 ± 0.07 / 1 ± 0.1 / 1.0 ± 0.1 / 1.2 ± 0.2 / 1 ± 0.08
At3g61970 / 1.1 ± 0.1 / 0.8 ± 0.06 / 1 ± 0.07 / 1.0 ± 0.1 / 1.1 ± 0.07 / 1 ± 0.06
At5g21105 / 0.9 ± 0.3 / 0.9 ± 0.05 / 1 ± 0.04 / 1.3 ± 0.1 / 1.2 ± 0.1 / 1 ± 0.1
At4g30250 / 1.3 ± 0.2 / 1.7 ± 0.2 / 1 ± 0.1 / 1.4 ± 0.09 / 1.9 ± 0.08 / 1 ± 0.1
At1g21670 / 1.4 ± 0.1 / 1.4 ± 0.08 / 1 ± 0.06 / 0.5 ± 0.08 / 0.7 ± 0.04 / 1 ± 0.1
At4g38400 / 1.6 ± 0.1 / 1.5 ± 0.1 / 1 ± 0.05 / 0.6 ± 0.04 / 0.7 ± 0.06 / 1 ± 0.03
At1g56020 / 1.6 ± 0.09 / 1.7 ± 0.2 / 1 ± 0.09 / 0.9 ± 0.1 / 1.3 ± 0.2 / 1 ± 0.03
At4g29720 / 1.1 ± 0.1 / 0.9 ± 0.009 / 1 ± 0.09 / 1.8 ± 0.2 / 1.5 ± 0.1 / 1 ± 0.2
At4g28720 / 1.5 ± 0.3 / 1.2 ± 0.1 / 1 ± 0.2 / 1.9 ± 0.2 / 1.8 ± 0.3 / 1 ± 0.04
At2g35700 / 1.1 ± 0.04 / 1.2 ± 0.08 / 1 ± 0.1 / 2.4 ± 0.07 / 1.5 ± 0.05 / 1 ± 0.09
Values are relative expression values representing the mean of four biological replicates ± standard error of the mean.
Supplementary TableV. qRT-PCR detected expression of REM1 and FLC in different genotypes
Genotype / REM1 / FLCCol / 1 ± 0.2 / 1 ± 0.1
Ws / 5.0 ± 2 / 0.20 ± 0.02
rem1-3 / 1.1 ± 0.1
sty1-1 / 0.47 ± 0.1 / 0.96 ± 0.09
sty1-1 sty2-1 / 0.29 ± 0.07 * / 0.91 ± 0.3
quintuple / 0.30 ± 0.03 * / 1.1 ± 0.2 **
35Spro:STY1:SDRX / 0.62 ± 0.1 / 0.82 ± 0.1
Values are relative expression values representing the mean of four biological replicates ± standard error of the mean. The tissues used were arial parts of 14-days-old plants. rem1-3, sty1-1, sty1-1 sty2-1 and 35Spro:STY1:SRDX should be compared with Col and SHI/STY- family quintuple mutant with both Col and Ws. Stars indicate a significant difference from wild type (Col and/or Ws) in a Student’s t-test: *, P<0.05; **, P<0.01.
Supplementary table VI. Mutant lines
Gene / Allele / Collection / Position / Accession / ReferenceORA59 / ora59-1 / GABI KAT / 1st exon / Col
ERF15 / erf15-1 / GABI KAT / Promoter / Col
YUC4 / yuc4-1 / JIC SM / 1st exon / Col / Cheng et al., 2006
YUC8 / yuc8-1 / JIC / 1st exon / Col
ERF38 / erf38-1 / Wisconsin / Downstream transcript / Ws
NGA2 / nga2-2 / AMAZE / 1st exon / Col / Trigueros et al., 2009
BLH11 / blh11-1 / GABI KAT / 1st exon / Col
BLH7 / blh7-1 / Syngenta / 4th exon / Col
REM1 / rem1-2 / FLAG / 6st exon / Ws
REM1 / rem1-3 / GABI KAT / 5’UTR / Col
Supplementary table VII. qRT-PCR primers
ACT_F1 / GTATGCTCTTCCTCATGCTATCCTT
ACT_R2 / TTCCCGTTCTGCGGTAGTG
YUC8_F1 / TGAAACAAAACAACCCACGA
YUC8_R1 / TTGATTCGCTTTGGGTCTTC
At2g31230_F1 / TTTCGCCACAAAAGGATCTC
At2g31230_R1 / GAGGATCGCTTTTTCCCTCT
At2g36630_F1 / TGGCAACACTGAATGTATCTGTGAA
At2g36630_R1 / GCGCTAGCAACCTGTGGGATGA
At4g31610_F1 / AGGAAATTTACAGCGCTTGCTACCA
At4g31610_R1 / ATTCCCCATATCCTCTCCCCTAAGG
At2g35700_F1 / GGCAGCTTTAGCCATCAAAG
At2g35700_R1 / CGGCGAAGACGTCTCTACAT
At3g51660_F1 / TCGGACGACCTCAAAACTTA
At3g51660_R1 / AACCAACGGTCGCTATGAGT
Sub_B-3_2F / CCGAGATAAGAGACTCAACGA
Sub_B-3_2R / CGGAGAGATTCTTCAACGAC
TOL1F / CAATGGCTTCGAGAGGAGAG
TOL1R / ACTGGAGGAGAGAACCAGCA
BELL1HOMO_F1 / CGAGCTTGGCTCTTTCAACAT
BELL1HOMO_R1 / GACGCCAACACCAGCTTCTC
At4g29720_F1 / CGGCGGAGAGTGAGTATCAG
At4g29720_R1 / GAAGCACAGATGCCAAATGA
At4g32800_F1 / CTCATCGTCGTCTTGGTCCT
At4g32800_R1 / CTTCCGTACCGCTTAACCAC
At4g30250_F1 / CAAAGCCCTTGTCCTAGATTCTT
At4g30250_R1 / TCCTCGTTTCTCCGACGA
At5g21105_F1 / GGCATGGAATCCGTCAGT
At5g21105_R1 / GGGTTAATTGCGCATTGAGT
At1g56710_F3 / AAGGCTCTGGTGTTATCAACG
At1g56710_R3 / CTTGTGTGGTTTGCAAGGAA
At2g46660_F1 / GCCGTTCTTTGGGAGATGAT
At2g46660_R1 / CCACAGCCGTTAGATATGGA
At4g38400_F1 / CGCAGGCTACGACGGCAAAAT
At4g38400_R1 / TCGTCGCATGGATCACAACCTTC
At1g21670_FI / ACGGAAACTGGAACGATACG
At1g21670_R1 / GGATGCATCGATACCAAACC
At1g56020_F1 / TTGGCCAGCTTTTCTCATCTTCGTC
At1g56020_R1 / GGTTCTTGCTTCCGTTGCTCTGTAA
Supplementary figure 1. qRT-PCR analysis of gene expression in different genotypes. (A) qRT-PCR analysis of the transcript level of ORA59 in XVEpro:ORA59 seedlings after 18 hours of treatment with mock or EST. Shown are the average of three biological replicates (three technical replicates for each biological sample), error bars represent standard error of the mean. (B) qRT-PCR analysis of the transcript level of ERF15 in three erf15-1 and one wild type plant. Shown is the average of three technical replicates, error bars represent standard deviation. (C) qRT-PCR analysis of the transcript level of ERF38 in wild type and erf38-1 plants. Shown is the average of two biological replicates (three technical replicates for each biological sample), error bars represent standard error of the mean.
Supplementary figure 2. The root length is increased in blh11-1 and blh7-1 seedlings. Length of the primary root of 10 DAG seedlings. Values represent the mean of three biological replicates (11-14 seedlings per replicate). Error bars indicate standard error of the mean.