Supplemental Figure

Supplemental figure

Supplementary Figure 1. Hierarchical tree of expression profiles under various stresses. The genes with missing values in one or more stress treatment were eliminated for clustering analysis. The folds of expression level changes (log2-transformed) by stress were subject to hierarchical clustering with Treeview program.FL: flag leaf; D: drought; S: salt. The original signal values were extracted from the published microarray data (Zhou et al 2007).

Supplementary Table 1 General information ofONACgenes with tissue specific expression patterns

LOC ID / Expression pattern / Real-time PCR Primers / cis-element name / cis-element sequence / Stimulus/tissue
Os01g29840 / Immature endosperm specific / F: 5′-GCCGATGCGCGAGTGT-3′
R: 5′-GCAATGCAAAAGCCATGATG-3′ / 2SSEEDPROTBANAPA / CAAACAC / Seed
AACACOREOSGLUB1 / AACAAAC / Endosperm
AMYBOX2 / TATCCAT / Seed
CAATBOX1 / CAAT / Seed
CANBNNAPA / CNAACAC / Seed
DPBFCOREDCDC3 / ACACNNG / Endosperm; seed
EBOXBNNAPA / CANNTG / Seed
MYB2CONSENSUSAT / YAACKG / Seed
MYBPZM / CCWACC / Seed
RYREPEATBNNAPA / CATGCA / Seed
SEF3MOTIFGM / AACCCA / Seed
Os02g12310 / Immature endosperm specific / F: 5′-TGCTGTGTAGGATCTCCAACAAG-3′
R: 5′-CACTGAAGAGAACTGCAACAAGCT-3′ / -300CORE / TGTAAAG / Seed; endosperm
CAATBOX1 / CAAT / Seed
CAREOSREP1 / CAACTC / Seed
DPBFCOREDCDC3 / ACACNNG / Embryo; seed
EBOXBNNAPA / CANNTG / Seed
MYCCONSENSUSAT / CANNTG / Seed
TATABOX1 / CTATAAATAC / Seed
Os02g15340 / Callus specific / F: 5′-GCGGCTCGCAGAACATG-3′
R: 5′-CCGCTCATCAGCAGCATCT-3′ / ACGTATERD1 / ACGT / Etiolation-induced
GT1CONSENSUS / GRWAAW / Light
NTBBF1ARROLB / ACTTTA / Auxin; meristem
Os06g33940 / Callus specific / F: 5′-GGCTTTGAAACCTAATTATACATGCA-3′
R: 5′-GCCAGCAATTATGAAGCATCCT-3′ / GT1CONSENSUS / GRWAAW / Light
IBOXCORE / GATAA / Light
Os05g34310 / Immature endosperm specific / F: 5′-CGAGTGGGTGCTATGCAAGA-3′
R: 5′-TTGATGGTGGTGCGGTGTT-3′ / CAATBOX1 / CAAT / Seed
CAREOSREP1 / CAACTC / Seed
DPBFCOREDCDC3 / ACACNNG / Embryo; seed
EBOXBNNAPA / CANNTG / Seed
GCN4OSGLUB1 / TGAGTCA / Embryo; seed
MYBPZM / CCWACC / Seed
MYCCONSENSUSAT / CANNTG / Seed
NTBBF1ARROLB / ACTTTA / Tissue-specific
PYRIMIDINEBOXOSRAMY1A / CCTTTT / Embryo; seed
Os11g03370 / Callus and root specific / F: 5′-TCATGACATTTTGCTTCGTGAGT-3′
R: 5′-AAATATGTGCCGCAAGAGAACA-3′ / ASF1MOTIFCAMV / TGACG / Auxin; SA; Light; root; leaf
CPBCSPOR / TATTAG / KT
ROOTMOTIFTAPOX1 / Root
Os07g37920 / Stamen specific / F: 5′-ACCCCATTCAACTGGTTCGA-3′
R: 5′-CCTGCTTAGCCCACTAGGAAAC-3′ / GTGANTG10 / GTGA / Pollen
Os11g31340 / Immature endosperm specific / F: 5′-CTGCCACCGCACAGGAA-3′
R: 5′-CAAGTGGGCCATGAGTTTGTC-3′ / CAATBOX1 / CAAT / Seed
EBOXBNNAPA / CANNTG / Seed
MYB2CONSENSUSAT / YAACKG / Seed
MYCCONSENSUSAT / CANNTG / Seed
Os11g31330 / Immature endosperm specific / F: 5′-GATCCATCCTCGCCCCATA-3′
R: 5′-GCAGACTCATCACTCCCTTCCT-3′ / -300ELEMENT / TGHAAARK / Seed
AACACOREOSGLUB1 / AACAAAC / Seed; endosperm
CAATBOX1 / CAAT / Seed
CANBNNAPA / CNAACAC / Seed
DPBFCOREDCDC3 / ACACNNG / Embryo; seed
EBOXBNNAPA / CANNTG / Seed
MYCCONSENSUSAT / CANNTG / Seed
PROLAMINBOXOSGLUB1 / TGCAAAG / Seed; endosperm
Os11g31360 / Immature endosperm specific / F: 5′-TCGCCGTTCCTGCCATT-3′
R: 5′-CATCTATGCCCTGGGAAGTCA-3′ / CAATBOX1 / CAAT / Seed
CAREOSREP1 / CAACTC / Seed
EBOXBNNAPA / CANNTG / Seed
MYB2CONSENSUSAT / YAACKG / Seed
MYCCONSENSUSAT / CANNTG / Seed
TATABOX1 / CTATAAATAC / Seed
TGACGTVMAMY / TGACGT / Seed
Os11g31380 / Immature endosperm specific / F: 5′-GGGATCGCACATTGAGTCATC-3′
R: 5′-TCACTTCCCTCTGCCAAAGAG-3′ / CAATBOX1 / CAAT / Seed
EBOXBNNAPA / CANNTG / Seed
MYCCONSENSUSAT / CANNTG / Seed
RYREPEATBNNAPA / CATGCA / Seed
TATABOX1 / CTATAAATAC / Seed
Os12g22630 / Stamen specific / F: 5′-CTCCTCCCATGGCACAAAAC-3′
R: 5′-AAATTTCCTCGTCAATCTCATCATC-3′ / GTGANTG10 / GTGA / Pollen
POLLEN1LELAT52 / AGAAA / Pollen

Note: The expression pattern of each gene based on microarray analysis and real-time PCR primers used for confirming the chip data are presented. F: forward. R: reverse.Putative tissue-specific cis-acting regulatory DNA elementsin the promoter regions of the OsNACgenescharacterized by PLACE (Higo et al. 1999) were listed.

Supplementary Table 2 General information of stress-induced OsNAC genes

LOC ID / Stress Responsiveness / Real-time PCR Primers / cis-element name / cis-element sequence / Stimulus / TF
Os03g21030 / D, S, C / F: 5′-CCTTCCGATCGAAGGCTTCT-3′
R: 5′-CTGGAGGTCTGGATGGATGATAA-3′ / ABRELATERD1 / ACGTG / ABA
Os12g29330 / D, S / F: 5′-GCAGGGTTAAGCTTTGTTTGTAAGT-3′
R: 5′-CTCTGTCTTGTCCTCCTGCATTT-3′ / MYB1AT / WAACCA / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os06g46270 / D, S / F: 5′-ACAATATGGGTAGGGCGATCAA-3′
R: 5′-TCCCTCTTCACATTGCCTTCA-3′ / MYCATRD22 / CACATG / ABA, drought / MYC/bHLH
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os07g48550 / D, S, C / F: 5′-AATTAAGGTCTCGGTTGGTTGATT-3′
R: 5′-CATACACAGAAAGAGGCCACATG-3′ / ABRERATCAL / MACGYGB / ABA
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os11g03370 / S, C / F: 5′-TCATGACATTTTGCTTCGTGAGT-3′
R: 5′-AAATATGTGCCGCAAGAGAACA-3′ / MYBCORE / CNGTTR / Drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os05g34830 / D, S / F: 5′-CCGGGAGGGATGAATGATCT-3′
R: 5′-CCATCTGACGGCTACCAATAACT-3′ / CBFHV / RYCGAC / Cold / CBF/DREB
CRTDREHVCBF2 / GTCGAC / Cold / AP2/CBF
DRECRTCOREAT / RCCGAC / ABA, drought, cold, light / CBF/DREB
LTRECOREATCOR15 / CCGAC / ABA, drought, cold / CBF/DREB
MYB2CONSENSUSAT / YAACKG / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os12g03050 / D, S, C / F: 5′-ATGCTGCTGCGTTCTGAGAA-3′
R: 5′-ATGTCCATGTTCAGTACGTAATTCAGT-3′ / MYB1AT / WAACCA / ABA, drought / MYB
MYBATRD22 / CTAACCA / ABA, drought / MYB
Os01g64310 / D, C / F: 5′-GGAAGTTCCATGCAGCAACTG-3′
R: 5′-TCGTCCCACTGGTCCCAAT-3′ / CBFHV / RYCGAC / Cold / CBF/DREB
MYB1AT / WAACCA / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os02g36880 / D, S / F: 5′-AGGACACCGGCCTCACGT-3′
R: 5′-GATCCATGCGTTGCTGCC-3′ / MYB1AT / WAACCA / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os01g15640 / D, S, C / F: 5′-GGAAGTTCCATGCAGCAACTG-3′
R: 5′-TCGTCCCACTGGTCCCAAT-3′ / ABRELATERD1 / ACGTG / ABA
ABRERATCAL / MACGYGB / ABA
ABREZMRAB28 / CCACGTGG / ABA, cold / CBF/DREB
ACGTABREMOTIFA / ACGTGKC / ABA
DPBFCOREDCDC3 / ACACNNG / ABA / bZIP
MYB1AT / WAACCA / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os11g03300 / D, S, C / F: 5′-CACAACACCTCATCCAAATTA-3′
R: 5′-TGCTGCCATCATCAAAGGAA-3′ / MYCATRD22 / CACATG / ABA, drought / MYC/bHLH
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os07g12340 / D, S, C / F: 5′-TGTGACTGCACCACTGTATATAAATTCA-3′
R: 5′-AAACTAACCTGCATATCATACTGCTCAT-3′ / ABRERATCAL / MACGYGB / ABA
CBFHV / RYCGAC / Cold / CBF/DREB
DRE2COREZMRAB17 / ACCGAC / ABA, drought
DRECRTCOREAT / RCCGAC / ABA, drought, cold, light / CBF/DREB
LTREATLTI78 / ACCGACA / Cold
LTRECOREATCOR15 / CCGAC / ABA, drought, cold, light / CBF/DREB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os01g60020 / D, S, C / F: 5′-CGGAAATAGGAGTGATGGCTAGA-3′
R: 5′ ACCACATTTGCAGAATCATGCT-3′ / ABREATRD22 / RYACGTGGYR / ABA, drought
ABRELATERD1 / ACGTG / ABA
ABRERATCAL / MACGYGB / ABA
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os06g04090 / D, C / F: 5′-AGTGGTCGAGCGAGTTGTTGT-3′
R: 5′-CCACGCCGTACGTCCTTT-3′ / MYB1AT / WAACCA / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
SBOXATRBCS / CACCTCCA / ABA
Os01g48130 / D / F: 5′-CCAGGCGGCTCCTAACAA-3′
R: 5′-CCTCAAAGTTAGGCATCGTCAA-3′ / LTRE1HVBLT49 / CCGAAA / Cold
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os02g34970 / D, S / F: 5′-TGTAATTAGTTATCCCCCAACGAGTA-3′
R: 5′-CGACTTGTGCTTCACACATATGC-3′ / MYB2CONSENSUSAT / YAACKG / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os07g37920 / D, S / F: 5′-ACCCCATTCAACTGGTTCGA-3′
R: 5′-CCTGCTTAGCCCACTAGGAAAC-3′ / ABRELATERD1 / ACGTG / ABA
ABREMOTIFAOSOSEM / TACGTGTC / ABA
ABRERATCAL / MACGYGB / ABA
ACGTABREMOTIFA2OSEM / ACGTGKC / ABA
ACGTABREMOTIFAOSOSEM / TACGTGTC / ABA
CBFHV / RYCGAC / Cold / CBF/DREB
DPBFCOREDCDC3 / ACACNNG / ABA
DRE2COREZMRAB17 / ACCGAC / ABA, drought
DRECRTCOREAT / RCCGAC / ABA, drought, cold, light / CBF/DREB
LTRE1HVBLT49 / CCGAAA / Cold
LTREATLTI78 / ACCGACA / Cold
LTRECOREATCOR15 / CCGAC / ABA, drought, cold / CBF/DREB
MYB1AT / WAACCA / ABA, drought / MYB
MYB2CONSENSUSAT / YAACKG / ABA, drought / MYB
MYBCORE / CNGTTR / Drought / MYB
Os07g48450 / D / F: 5′-CGAATTTACGGATGCTACGTACTG-3′
R: 5′-TCGTCGTCCATGCTTCCAT-3′ / MYB2CONSENSUSAT / YAACKG
MYCATRD22 / CACATG / ABA, drought / MYC/bHLH
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os12g41680 / D / F: 5′-TGCCTGAATTTGGCAGCTT-3′
R: 5′-ACACCTCACCCCCAATGGA-3′ / MYB1AT / WAACCA / ABA, drought / MYB
MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH
Os05g10620 / D, S, C / F: 5′-GGCGCTATTCCGAAGTGAAC-3′
R: 5′-GCGCTGGTGAGGCCAAT-3′ / MYCCONSENSUSAT / CANNTG / ABA, drought, cold / MYC/bHLH

Note: The stress-inducible expression pattern of each gene based on microarray analysis and real-time PCR primers used for confirming the chip data are presented. D: drought; S: salt; C: cold. F: forward. R: reverse. The stress response-related cis-acting regulatory DNA elements in the promoter regions of the OsNACgenes characterized by PLACE (Higo et al. 1999) were listed.

Supplementary Table 3 List of primers used for Y1H plasmid construction

Construct / MEME group of NAC gene / Primer sequences
pGAD-Os01g64310 / A / F: 5'-CAGGAATTCCAACTCGCTGGCAATATGG-3'
R: 5'-CAGGGATCCCAGTTAGCACTACGAGCAAGA-3'
pGAD-Os02g36880 / A / F: 5'-CAGGAATTCAAGATGCGATTAGCCAGG-3'
R: 5'-CAGGGATCCATTTCAATAGGCCCAGAG-3'
pGAD-Os12g41680 / C / F: 5'-CAGGAATTCTCGATGTCGATGATGAGCTT-3'
R: 5'-CAGGGATCCCAACGCAAACCTCTACTTCTG-3'
pGAD-Os12g03050 / D / F: 5'-CAGGAATTCGCAGCTAGCATGGTGGAGTCTAC-3'
R: 5'-CAGGGATCCGAAGAACTAAATAATGAAGCAGTC-3'
pGAD-Os03g21030 / E / F: 5'-CAGGAATTCAAAGGAGGGAGCTTTGAAGG-3'
R: 5'-CAGGGATCCGCATCAACAAACTGCCACTAA-3'
pGAD-Os01g48130 / H / F: 5'-CAGGAATTCTCTCTCGTCTTGTTGGATTATACAG-3'
R: 5'-CAGGGATCCATCCATCAGGGGCCAAAG-3'
pGAD-Os06g15690 / J / F: 5'-CAGGAATTCCTAGTAGGGAGGGGAGGT-3'
R: 5'-CAGGGATCCCGCAGAAGGAAGAGGCT-3'
pGAD-Os08g42400 / L / F: 5'-CAGGAATTCATTGATACGATGGAGAGGGC-3'
R: 5'-CAGGGATCCCATTGATGCAGATCACTTTCA-3'
pGAD-Os01g70110 / L / F: 5'-CAGGAATTCTTGACCAACAAACCCAACC-3'
R: 5'-CAGGGATCCGACATTAGTAGAGCAAAAGAAGG-3
pGAD-Os02g12310 / N / F: 5'-CAGGAATTCATCATCATCTTCCATAGCTGTAC-3'
R: 5'-CAGGGATCCTTTAACTCAAATCTCCCGC-3'
pGAD-Os01g09550 / O / F: 5'-CAGGAATTCGGCATGAATAGGGGGCAC-3'
R: 5'-CAGGGATCCTGGTTAGTTTTCTTTTGGTATTG-3'
pGAD-Os10g21560 / O / F: 5'-CAGGAATTCTCCGTGTGGACAATCATGTC-3'
R: 5'-CAGGGATCCCGACCAGTCCGTATCAAAATAG-3'
pHIS2-OsERD1 / F: 5'-AAGAGCACAAGCAAAACGC-3'
R: 5'-CGAAGGAAACACGCAAGG-3'
pHIS2-OsOAT / F: 5'-GCAGAATTCCTCACATGTTATAGATACGATGTG-3'
R: 5'-GCAGAGCTCTGAAATGGGGTTTAACTCTACT-3'