Figure S1
Figure S1. Modular structure of rice proteins encompassing the SPX
domain. Different colors indicate the amino acid ranges of the domains.
Purple, SPX domain; blue, EXS domain; yellow, MFS domain; red, RINGtype Zinc finger domain; pink, no conserved domain.
Figure S2
Figure S2. Expression analysis of OsSPX-MFS1,
OsSPX-MFS2 and OsSPX-MFS3 in leaves and roots.
Figure S3
Figure S3. Expression analysis of OsSPX-MFS1, OsSPX-MFS2, OsSPX-MFS3 and
osa-miR827 under different Pi starvation treatments. Seeds were geminated and
growth on nutrient solution with different Pi concentration for three weeks. Roots
were sampled separately for analysis. All data are means of three biological
replicates with error bars indicating SD. Expression of OsACTIN was used as the
internal control. nd indicates non-detectable.
Figure S4
acctctctagttcattttgtgagagagctctacccagctccgctcccatattagatagaactgaaactgtttggttga
gcttcagctccaggagagatggagctagagctggagccgtgccaaacaggaccatctaggtactgacacgtgta
G-box
gaaaatacttcccggagctttggttcttctgttccactccacccatcggcacaggcgggcgtctgcactctgggcctc
tggcgtgcggcgcccgcgcgtggtctggttgcgcagcgcgctgcgccgggtggaatccgttgcggcggcgtacac
cgtcacggcgtcacacactcacaccagcgccgcgcgccgcgggggacgtcagggactcgcgcgggccatgcag
gttgggctggctctcggcctctcgcctgtatctcaagggggtagcagcccattgcgtcggtagacctaggactcccc
ccgtgggccggtcacgtggagtcgtacgtggccctcgtcgatagcgccgcaggcctctctcaccacaagtccacg
G-box
atgctgccccgaggatggttgtacacatggtagaacagtccgagaggggaacaattttgcctacaaattatccaca
E-box
actttgcgagcgtttcatccgttccgaacagtccccttcctctgctgccccatcatcgctcatatctggattttttttttg
aaaggagtatatctggatatatataataccccttcgtactcgtaaagaaaattgtttaggacaatgtttaagtcaga
ccttgaaaatataaatcataaataactctcaaattgttgagtttgaaaatataaaaattatataaatagatttgtctt
TATA box
TATA box
gaaaaatattttcataaaagtatacatataatactttcaataaatatttttttatagaaacaataagtaaaagttgtat
tttagagaccgtgtcactgtccaaaatgactttctctacgagtaagagagagtattcctttttttttcatgactggaca
W-box
W-box
accgcaaaatccacgcgttcgtccgctggtacaactcctccctgtcacggctaatttaccaagaccgttgatccaaa
tctacgctcatcatctccacctagcagaagaggcagtacagtcagccagtggcacagcgtcagcgtcactacaga
gtgtgcacgtacatacatgttgctgactgtctcctgcttaattacttgatgcttccatgtcgtcgtgcttctgcggggg
W-box
acggtttattcctcgcgaatatgcctctaattcgctggcatcaacgtaccatcatcccttccgtgcacgcacctaccc
P1BS
gatccaaaatatatccttcgcttcaatacagcactagcagcttcatctcattcgccccatcttccagtcttccaggcgt
ccagctccagtttcttctcattgacaatatatccagattatgctctgggcttgctaattgctactagtagaatactagg
ggtggtcgtggtgtgcaggtgagattattgctacCCATGAACCTGTTTTGTTGCTGGTCAT
CTAGCTACCCGTGCATGCCTGGAGATTGGAGAATAATTGACGATGCA
GCAGTCGGCTTATTGGCTCTTGGGCACGCGTGGTTAGATGACCATCA
GCAAACAAGTTCGTGAG
Figure S4. Nucleotide sequence of the promoter and gene region of osa-miR827.
The letters in capital indicates the precursor sequence of osa-miR827. The ciselements of the promoter region were colored in red.
Figure S5
Figure S5. Expression analysis ofOsSPX-MFS1 (A), OsSPX-MFS2 (B),
OsSPX-MFS3 (C) and Pri-miR827 (D) in osa-miR827 overexpression
lines and msf1in the roots. Seeds were geminated and growth on
nutrient solution with high Pi concentration (HP, 10 mg/L) and low Pi
concentration (LP, 0.1 mg/L) for three weeks. Total RNA was extracted
from the roots and used for qRT-PCR. All data are means of three
biological replicates with error bars indicating SD. Expression of
OsACTIN was used as the internal control. WT, wild type; Oe1/2/3, osamiR827 overexpression lines; mfs1, OsSPX-MFS1 mutant line NC2634.
Table S1. Sequences of the primer sets used in the study
Primer Name
OsSPX-MFS1
OsSPX-MFS2
OsSPX-MFS3
Pre-miR827
OsSPX1
OsIPS1
OsPAP10
OsACP1
OsPT2
OsPT3
OsPT7
OsPT8
Tos17-3'
NC2634
OsSPX-MFS1:Pro
miR827-Oe
Forward（5` → 3`）
GCGATTCTTGGGTGTACTGT
CCTCCTGAATGTAACCCTG
CGATTGGTTCAGCATACAGA
TCTAGCTACCCGTGCATGCC
GACCAGCTTCTACCATCAAACG
AAGGGCAGGGCACACTCCACATTATC
ATACTGGCAGCCGACGGATGA
AGATGGAGGAGACGCTGCTG
CACAAACTTCCTCGGTATGCT
AGACGGTGGTTCAACAGAG
CTTGCTCGGCTTCCTCCT
CCTACTTGTGTTTGTCTATGTG
ATTGTTAGGTTGCAAGTTAGTTAAGA
GTTTTGCGGTTCTGCTTCTC
ACTCCACAAGGATTGGCTCCG
GATCGGATCCGGGTGGTCGTGGTGTGCAGGT
Reverse（5` → 3`）
CGTGAAAGCAACGACAGGTT
AGGAACTGTGTCAACTGCTT
GTGCTGAGACGACATACTGG
GCGTGCCCAAGAGCCAATAA
AGTTCCTGCTGCTCCTCTGG
ATTAGAGCAAGGACCGAAACACAAAC
GAGGGAGCTGGAGCGGAGAA
TCGCAATCCTCACACACACAT
GAAACCCCACAAATCCACAAC
GCAAACCAAACTAACAAAATACC
GCCATTATTTCATCATCCAGC
GTGCCAAATTGCTGGTCTG
TCCACCAAATCCTGACCAAC
CTTGTCGTTCAATACCTGTATC
GATCGGATCCCAAATGAACCAGAGAATCGAC
Table S2. Information of OsSPX-MFS genes in rice
Gene name
Gene locus
Protein length
cDNA accession No.
Total EST
OsSPX-MFS1 Loc_Os04g48390
696
AK066068
9
OsSPX-MFS2 Loc_Os02g45520
697
AK241359
18
OsSPX-MFS3 Loc_Os06g03860
698
AK243620
182
OsSPX-MFS4 Loc_Os09g34990
570
NF
NF