Fig. S1
Fig. S1 Splicing variants of HRE1. a Schematic maps of the HRE1 gene and its splicing variants, HRE1α
and HRE1β transcripts. AUG and UGA* indicate start codon and stop codon, respectively. b The amino
acid sequence alignment of HRE1α and HRE1β. AP2/ERF domains of both HRE1α and HRE1β
transcripts were presented. Alignment was made by ClustalW using default parameters.
Fig. S2
Fig. S2 Expression analysis of HRE1α under abiotic stress conditions. Quantitative RT-PCR analysis of
HRE1α under 300 mM NaCl (a), 300 mM mannitol (b), 100 μM ABA (c), or 10 µM MV (d) for the
indicated times. GAPc was used as an internal control. Transcript levels at 0 h were set to 1. Reactions of
each technical replicate were performed in triplicate. Two technical replicates were measured for each
biological replicate. Data shown are the means ± SD (n = 6). Similar results were obtained from at least
two biological replicates, with one shown here.
Fig. S3
Fig. S3 Generation of HRE1α OXs. Schematic map of plant expression vector for overexpression of
HRE1α, and expression analysis of HRE1α in HRE1α-overexpressing T1 lines by semi-quantitative RTPCR. Circled lines were selected for further analysis. GAPc was used as an internal control.
Table S1 Primers used in this study
Gene
Forward
Reverse
HRE1α
5′-CTTTATCCATGGCTAGGCTC-3′
5′-ACTCCTTTGATTGGATCACG-3′
GAPc
5'-GTGTCCCAACCGTTGATGTC-3'
5'-TCCCTTGAGTTTGCCTTCGG-3'
HRE1α
5'-CTTTATCCATGGCTAGGCTC-3'
5'-TCAAGACAGCTACTACTAGG-3'
GAPc
5'-CACTTGAAGGGTGGTGCCAAG-3'
5'-CCTGTTGTCGCCAACGAAGTC-3'
HRE1 promoter
5'-GCGAAGCTTTTGATAATTAAAAAGAAAAT-3'
5'-GCGCTGCAGAGAACCAAACGATGAACATA-3'
Cloning
HRE1α OX
5'-CGCGTCGACATGTCTCAAAGCTTTGAACT-3'
5'-GATGGATCCTCAGGACCATAGACCCATGT-3'
Cloning
sGFP
5'-ATCCTGCAGATGGTGAGCAAGGGCGAGGAG-3'
5'-ATCGTCGACCTTGTACAGCTCGTCCATGCC-3'
Cloning
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-CAGGTCGACTCAGGACCATAGACCCATGT-3'
Cloning
HRE1α N160
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-CGCGTCGACGTTGGCTTCTTCACTATCAT-3'
Cloning
HRE1α N145
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-CACGTCGACGCCCATGATGATATCTTCGA-3'
Cloning
HRE1α N136
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-CGCGTCGACTGTATCAATCAGAAGCGTGT-3'
Cloning
HRE1α N130
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-ATAGTCGACGTCCGGATTATTCTCCTCCC-3'
Cloning
HRE1α N120
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-ACAGTCGACAAGACAGCTACTACTAGGCG-3'
Cloning
HRE1α N87
5'-CAGGAATTCATGTCTCAAAGCTTTGAACT-3'
5'-ACAGTCGACCTTTCCAGAGGATTCGTTAG-3'
Cloning
5'-GCGCTCGAGTTGAGACTTTTCAACAAAGG-3'
5'-CACGTCGACGTCCTCTCCAAATGAAATGA-3'
Cloning
5'-GCGCTGCAGTTATTGTTCATTTTTGAGAA-3'
Cloning
5'-ATACCCGGGGAATTTCCCCGATCGTTCAA-3'
5'-GCGTCTAGACCGATCTAGTAACATAGATG-3'
Cloning
5'-CAAGAATTCGCAAGACCCTTCCTCTAT-3'
5'-ATACTGCAGCAGCGTGTCCTCTCCAAA-3'
Cloning
Firefly luciferase
5'-CGCGGATCCATGGAAGACGCCAAAAACAT-3'
5'-CAGTCTAGATTACACGGCGATCTTTCCGC-3'
Cloning
Nos terminator
5'-ATACCCGGGGAATTTCCCCGATCGTTCAA
5'-GTCGAGCTCGAATTTCCCCGATCGTTCAA
Cloning
HRE1α entire
ORF
CaMV 35S
promoter
Renilla luciferase
Nos terminator
CaMV 35S
promoter core
Purpose
Quantitative
RT-PCR
5'-CGCGTCGACAAAAATG
ACTTCGAAACTTTATGA-3'
Quantitative
RT-PCR
Semi-quantitative
RT-PCR
Semi-quantitative
RT-PCR