TAA
ATG
(a)
100 bp
UTR
Intron
T-DNA
40 cycles
Tubulin
35 cycles
Primer pairs
gr
f1
C
Promoter
1
GRF11
ol
-0
(b)
ORF
Supporting Information Fig. S1 Localization of the insert (a) and transcript
abundance (b) of the analyzed T-DNA insertion mutant lines.
1
Fig. S2 Expression analysis of AHA1 and AHA7 in wild-type (WT), grf11 mutant and
transgenic lines under different Fe status.
Seedlings were grown in +Fe medium for 6 wk and then transferred to nutrient media,
or deprived of Fe for 7 d. Expression levels were normalized to UBQ10 and then to
the value of each gene from the WT under Fe sufficient conditions. Data are means ±
SD (n = 3). *, Significant change vs +Fe; +, significant change vs WT +Fe.
2
Fig. S3 FCR activity and Fe content in GRF11 over-expression transgenic lines.
(a) Arabidopsis plants (wild type (WT), FIT-OX, GRF11-OX2, and GRF11-OX5)
were grown in +Fe medium for 6 wk and then transferred to nutrient solution (+Fe),
or deprived of Fe (-Fe) for 7 d. Data are means ± SD (n = 7). *, Significant change vs
+Fe.
(b) and (c) Arabidopsis plants (wild type (WT), FIT-OX, GRF11-OX2, and
GRF11-OX5) were grown in +Fe medium for 6 wk, then roots and shoots were
separately harvested for Fe content determination. Data are means ± SD (n = 4).
3
1
2
3
4
5
6
(a)
(b)
Fig. S4 Identification of interaction between FIT and GRF11 in yeast.
Yeast colonies were grown on synthetic dropout (SD)–Leu/–Trp (b) or
SD–Ade/–His/–Leu/–Trp plates (a) for 3 d. The numbers of 1 to 6 indicate plasmid
combinations as follows: 1, pAD-T with pBD-53; 2, pAD-T with pBD-Lam; 3,
pAD-FIT with pBD-GRF11; 4, pAD-FIT with pBD; 5, pAD with pBD-GRF11; 6,
pAD with pBD.
4
Table S1 Primer pairs used for qRT-PCR
Gene
Locus
Primers
IRT1
AtU27590
fw:5'-GCGACTTGTAGTGCGGCTATG-3'
rev:5'-CGTTGCACGAGCGATTCTG-3'
FRO2
At1g01580
fw:5'-GGAGAAGGTGTTGCTCCATCTC-3'
rev:5'-GGTTAGGCAAGTTTAAGCTCTG-3'
AHA1
At2g18960
fw:5’-GCAAGCCAAGAGAAGGGCCGAG-3’
rev:5’-GAGTCATGATGGGGTTGGTTTTCGG-3’
AHA2
At4g30190
fw:5'-TTTGCCGGAGTCTTCCCAGGT-3'
rev:5'-GGGGCATCATTGACACCATCACCAG-3'
AHA7
At3g60330
Fw:5’-AGGGCTAACGGGTGATGCTTTGG-3’
Rev:5’-TGCTCTCCGTGGTGATAGCCTTG-3’
FIT
At2g28160
fw:5'-GTATCAATCCTCCTGCTT-3'
rev:5'-TCTCGGTTACATCATCACT-3'
bHLH038
At3g56970
fw:5'-AGCAGCAACCAAAGGCG-3'
rev:5'-CCACTTGAAGATGCAAAGTGTAG-3'
bHLH039
At3g56980
fw:5'-GACGGTTTCTCGAAGCTTG-3'
rev:5'-GGTGGCTGCTTAACGTAACAT-3'
bHLH100
At2g41240
fw:5'-AACTCAGAGGAAGGGGTTACA-3'
rev:5'-GATGCATAGAGTAAAAGAGTCGCT-3'
bHLH101
At5g04150
fw:5'-CAGCTGAGAAACAAAGCAATG-3'
rev:5'-CAGTCTCACTTTGCAATCTCC-3'
5
EIN3
At3g20770
fw:5'-TCCTCAGCCAAGGCCAGTGA-3'
rev:5'-TGTGGGTTGAAGCAGTGACACG-3'
EIL1
At2g27050
fw:5'-TGGAGTTCCGGAAAACGGGCA-3'
rev:5'-GCTGCTGATTCTGAGCTGCTTTTGC-3'
GRF11
At1g34760
fw:5'-CTATCACTCAAGGCGTATG-3'
rev:5'-GTATGGCTCCGTAACTAAG-3'
UBQ10
At4g05320
fw:5'- GTCTTCGTGGTGGTTTCTAAATCT -3'
rev:5'- TTATTCATCAGGGATTATACAAGG -3'
6