Fig. S1 Inheritance of salt-resistant phenotype of AT1362 in T3 generation. a Germination of WT plants and three
independent T2 lines of AT1362 on 140 mM NaCl-containing medium. b Germination ratio of WT plants and
three independent T2 lines of AT1362 on 140 mM NaCl-containing medium up to 6 DAG.
Fig. S2 Comparison of amino acid sequences between AtSFT12 and other Qc-SNAREs in Arabidopsis. a
Phylogenetic tree of Qc-SNAREs in Arabidopsis. Phylogenetic tree was made by ClastalW2 Phylogeny using
default parameters. b Multiple alignment of SNARE motif of AtSFT12 with those of 11 other Qc-SNAREs in
Arabidopsis. Alignment was made using ClustalW2 with default parameters. Box indicates the 0-layer residue in
the center of SNARE motif.
Fig. S3 Oxidative stress responses of AtSFT12 OXs and atsft12-2 mutants. a and b Responses of WT and AtSFT12
OX seedlings to 0, 1.5, 2, or 2.5 μM MV. c and d Responses of WT and atsft12-2 seedlings to 0, 1, or 1.5 μM
MV. In b and d, error bars represent standard deviation (n = 24 plants).
Fig. S4 Salt stress response of atsft12-1. a Semi-quantitative RT-PCR analysis of AtSFT12 transcription levels in
4-week-old WT plants and atsft12-1 mutants. GAPc was used as an internal control. b and c Responses of WT
and atsft12-1 seedlings to 0, 130, or 140 mM NaCl. In c, error bars represent standard deviation (n = 24 plants)
and * indicate t-test P < 0.05.
Fig. S5 Expression patterns of AtSFT12 under abiotic stress conditions. Semi-quantitative RT-PCR analysis in
response to 300 mM NaCl (a), cold (b), 100 μM ABA (c), 300 mM mannitol (d), or 10 μM MV (e) treatment for
the indicated times. GAPc was used as an internal control. Similar results were obtained from at least two
biological replicates, with one shown here.
Table S1 List of primers used in this study
Gene
Forward
Reverse
Purpose
AtSFT12
5′-GTGGTTCTTCTCGTTACCTG-3′
5′-AGGCCAGATTTGAACTGGTC-3′
Quantitative
RT-PCR
GAPc
5′-GTGTCCCAACCGTTGATGTC-3′
5′-TCCCTTGAGTTTGCCTTCGG-3′
Quantitative
RT-PCR
AtSFT12
5'-AGAGAAGGGCTAAGCACTAG-3'
5'-ATGTTGTTCTTCACCCCTGC-3'
Semi-quantitative
RT-PCR
AtSFT12
mutant
5'-GTCTCACAGGGACTTCTTAG-3'
5′-AGGCCAGATTTGAACTGGTC-3′
Semi-quantitative
RT-PCR
GAPc
5'-CACTTGAAGGGTGGTGCCAAG-3'
5'-CCTGTTGTCGCCAACGAAGTC-3'
Semi-quantitative
RT-PCR
RD29A
5'-GAAACAGAGTCTGCCGTGAC-3'
5'-TGCTGCCTTCTCGGTAGAGA-3'
Semi-quantitative
RT-PCR
RAB18
5'-AATGCTTCACCGCTCCGGAT-3'
5'-TTCTTCTCGTGGTGCTCACC-3'
Semi-quantitative
RT-PCR
WRKY33
5'-AGATTGTGGGAGTGAACCTG-3'
5'-GACTGAAGACGAATCCTGTG-3'
Semi-quantitative
RT-PCR
AtSFT12
5'-ATAGGATCCATGGCGTCCAATCGCGGTGC-3'
5'-CGCGGTACCCTATCTTTTGAACATCTTGG-3'
Cloning
LB1
5′-ATGGTCATAGCTGTTTCCTGTGTGAAATTG-3′
TAIL-PCR
LB2
5′-AACCTGTCGTGCCAGCTGCATTAATGAATC-3′
TAIL-PCR
AD1
5′-NGTCGASWGANAWGAA-3′
TAIL-PCR
AD2
5′-TGWGNAGSANCASAGA-3′
TAIL-PCR
AD3
5′-AGWGNAGWANCAWAGG-3′
TAIL-PCR
AD4
5′-WGTGNAGWANCANAGA-3′
TAIL-PCR
LB-seq
5′-CATTAATGAATCGGCCAACG-3′
Sequencing