Article title: S-like ribonuclease gene expression in carnivorous plants

Journal name: Planta

Author names: Emi Nishimura, Minako Kawahara, Reina Kodaira, Marina Kume,

Naoki Arai, Jun-ichi Nishikawa and Takashi Ohyama*

*corresponding author

Affiliation and e-mail address of the corresponding author: Department of Biology,

Faculty of Education and Integrated Arts and Sciences, Waseda University, 2-2

Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan

E-mail: ohyama@waseda.jp

Fig. S1 Structures of D. muscipula dm-I (a) and C. follicularis cf-I (b) and their 5′- and 3′-flanking

regions. Nucleotide position numbers on the right are counted from the transcription start site (+1). The

deduced amino acid sequence is shown above the nucleotide sequence (typed in gray) and the putative

secretion signal peptide is boxed. The translation terminal codon is marked with an asterisk. Exons are

typed in red. Positions of hybridization by degenerate primers (Materials and methods) are indicated with

broken lines each with an arrowhead. PLACE (Higo et al. 1999)- and PlantCARE (Lescot et al.

2002)-based analyses identified the following elements in the 5′-flanking regions: TATA box; CAAT

box; P1BS, PHR1-binding sequence found in upstream regions of phosphate starvation responsive genes

from several plant species; W box, WRKY-binding site involved in transcriptional upregulation

responding to a diverse set of stresses, including infection by pathogens and wounding; GT-1 motif, plays

a role in pathogen- and salt-induced SCaM-4 gene expression; TC-rich repeats, cis-acting element

involved in defense and stress responsiveness; TCA element, cis-acting element involved in salicylic acid

responsiveness.

ATCTCGCTGCTAGGTGGAGATAATCATATATAGATAGATATATATATATATATATATATA M K N S V F I K L L V W Q S TTATACC ATG AAG AAC TCT GTG TTC ATC AAG CTT TTG GTT TGG CAA TCT L A A V A L C Q G F D F F Y F V CTA GCT GCA GTG GCT CTA TGC CAA GGT TTC GAC TTC TTC TAC TTC GTC Q Q CAA CAG GTGACTCACGTGCACCATTTACATGATACACCACCATGTTAACTCACGCCAT CATGTGTTCAGTAGCTCGTCGTTCGAACGGTTCAACCAAATTGATCGAACAGTTCGGTGG TCTTGATCGAACTGTTAGGTCAACTTAGCCGAACTGCTCGATTCGGCCGAAGTGTTTATA GTCAAGTAGTGAGTTTTTTCTTTGCATTGACTTTGTCTCTCATTTTGAATGTATATATAT W P G S Y C D T S Q S C C ACATGCAG TGG CCA GGA TCA TAC TGT GAC ACA TCG CAG AGT TGC TGC Y P T T G K P A S D F G I H G L TAT CCT ACA ACT GGG AAG CCG GCT TCG GAT TTT GGC ATC CAT GGG CTC W P N Y N S G S Y P S N C D S S TGG CCT AAC TAC AAC AGT GGT AGC TAT CCA TCG AAT TGC GAC TCC AGC N P F D P S Q AAC CCT TTC GAC CCG TCT CAG GTATCTATCTGCATATATACAAAAATGTATGTATG I Q D TAAGTACATCGTCGTGATCATATGACATATATAATCACTCTTATGCAG ATC CAA GAT L L S Q M Q T E W P S L S C P S CTG CTG AGC CAA ATG CAA ACC GAG TGG CCG TCA TTG TCC TGC CCA AGC S D G T S F W T H E W N K H G T AGC GAT GGT ACA AGC TTC TGG ACA CAC GAA TGG AAC AAG CAC GGG ACC C S E S V L N E H A Y F Q A A L TGC TCT GAG TCT GTG CTC AAT GAA CAC GCT TAC TTT CAA GCC GCT TTG S L K N S S N L L Q T L A N A AGC CTC AAG AAC TCG TCC AAC CTC CTC CAA ACC CTA GCT AAT GCA G G TAATTAATTAACTAGTACATATAGCAAACTAACTGCGGATGATCATCATTTCAAAGTTTT G I T P TAACACAATTATCGGCAGTATAAATATCAATGTGTACCGTGAACAG GA ATT ACT CCG N G N S Y N L S D V L A A M K Q AAT GGC AAC TCA TAT AAT TTA TCG GAC GTG TTG GCC GCG ATG AAA CAA A T G G Y D A Y I Q C N T D Q N GCA ACT GGA GGA TAT GAT GCT TAC ATC CAA TGC AAC ACC GAC CAG AAT G N S Q L Y Q V Y M C V N T S G GGA AAT AGC CAG CTC TAC CAA GTA TAC ATG TGT GTT AAC ACT TCC GGG Q S F I E C P V A P S Q N C N P CAA AGT TTC ATC GAA TGC CCA GTG GCT CCT AGT CAG AAC TGT AAC CCC S I E F P S F * AGC ATT GAG TTC CCT TCC TTC TAA CCCACCGGCCTCATGCTCATGAGATCTAGC CAGTGAAATGTTAGTTTCATTGTCAATCAGTCCAACAACAACAACAATAAACTAACTCAACAA TAAACATATGTTCTTGTATTGATATCAATAGATAGTATAGATAGATGGTGATTGACACAT GTATTTGGAATATGATGAATTGTTTTTCATCCAATTTTTACCTAGATTTTACAAACAAAT

TCAATAGCCT TGTCTTAATG TCTCATGACT CCGATCAACA CATATATATG CGGACAATGA GGTTAATTTA TTCATTTCGG AAATCGGTTG ACCTGCTGCC GGAGTATGGT CAAAAGCCCC CATAAGGTAA GACTGCCGAC TTGGCATGCA TGCCGCATGC GAATTTAAGT AATAAAAGAT CAACGATGCG GGTGAGTAAT AAACTAATAA TTATGATACC TCAGTAGAGT TCGACATTTG ATATAATTTA ACCAAAATTA AGGATTTTAT CTTTATCTGC AACGTTTAAT TAATATTTTT CAGGTTGAGA TAGATAAAAT GATCCGTTAA GATATAGTTG ACGTTGAACC AATATCGACA AATTAATTAA TGGTTTGGAG CAACCTAATT AACGTAATTA AATTGATCTA TATATGCCTA ACCATATATA TAGGTAACTT AGCTTATAGC TAGTATACTT GAAACGCTTG ATTACGATAA TTAATAAAAT GATGTGAATC ATGTGATCGT GTTAAATATG GAAAAGATCA CCTTTAATTC CCATTATATA GCCCCTGTAT CTGTACGTAT AATATCAAGC TATTCTTGAA TGGGATTAAC TAGCGAAATT AAGTCTTGTA AATTAATAAC ACGAAACTCT CGATTAATCA ATTAGCATAT TCCCCCCCTT GCTTGTTAAT TAACAGTATA TATATGGCTC CCCCTGCAAA GAGCAATCTC

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M K S K G S L ATCATGTATCACTCTCAAAGCCCTCGAATTTTGAAAAG ATG AAG TCC AAG GGT TCA CTC L I K L L I I Q Y L S S L F V A Q TTG ATC AAG CTT TTG ATA ATT CAG TAC TTG TCA AGT CTA TTT GTT GCA CAG N F D F F Y F V Q Q AAT TTT GAT TTC TTC TAC TTT GTT CAA CAA GTGAGTTTTTGAGTTTCTCCACAATGA GATTTCTATATTTTTTATCCACAATGAAATTTCCCTATTTCCCCATCTGTATCGAACTTAATAATCA W P G S Y C D T S TGATTCACCTACACTTTTTGATGTATAACAG TGG CCA GGA TCA TAT TGT GAC ACA TCG Q G C C Y P P T G K P A S D F G I CAA GGT TGC TGC TAT CCA CCT ACT GGG AAG CCA GCA TCG GAT TTT GGC ATT H G L W P N N N D G S Y P S N C D CAT GGG CTT TGG CCA AAC AAC AAT GAT GGA TCA TAC CCT TCA AAC TGT GAT P N N P F N Q S E CCT AAC AAC CCC TTC AAT CAA TCT GAG GTGATTTTAGCTCAAAATTGTTTCAGTAGAT AACATTAGCCCTTTTTTAGCGTCTTATTATGTGTTTTGGCTTTAAAATCATCGTCTACAAATCTAAT ATATGTACGTTAAAAAGTTTACTCTTGCTGACTACTTGCGTCACTCGAGTGTGTTTCTTGTTCTCCC TTTGATTTTTTTGTATATATTGTCATATAATCACATGTGTCACCATGTAAAGTGTCACCCTTTTACA ACATCATATAGAAAGTTACATACACTTGCCAGTTACATGAATGAGTTCAGCTGAAATGGCGCATACT GTTAACTTAAATCTTGAATATTGTATTGTCCTCTTATTAATTTGGTTGTGTTGTTTATCGTGGAATAG V S D L M S S L E T N W P S L A C GTA TCT GAC CTG ATG AGT AGC CTG GAA ACG AAT TGG CCA TCA CTA GCA TGC P S S D G I S F W T H E W D K H G CCG AGC AGC GAC GGC ATA TCC TTT TGG ACG CAC GAG TGG GAC AAA CAT GGT T C S E S V L D E H D Y F Q A A L ACT TGC TCA GAA TCT GTC CTT GAC GAA CAT GAT TAT TTT CAA GCA GCT CTC N L K N K T N L L Q A L A S A AAC TTA AAA AAT AAA ACC AAT CTC CTC CAA GCC CTT GCA AGC GCA G GTAAT TTTCAATTTAGTTCACTTCTTAAAGTAAATATTTTCGACATTGCTTGTGCTAGTAGTCAGTGTATTA G I N P N G E S ATTAATTCTAATAAAGCTAAAAAAATGTGTTGCAG GA ATA AAT CCA AAT GGG GAA TCG Y N L S D I I N A I N Q S D G F T TAC AAC TTG AGC GAC ATT ATT AAT GCT ATT AAT CAA TCA GAT GGG TTT ACT P S I E C N D D E S G N N Q L Y Q CCA TCG ATA GAG TGC AAT GAT GAT GAA TCA GGT AAC AAC CAG CTT TAC CAG V Y M C V D T S G S N L I E C S V GTT TAC ATG TGT GTA GAC ACT TCT GGA TCT AAC CTC ATC GAG TGC TCG GTG S P T G A N C A N Q I T F P A F * TCA CCT ACG GGA GCA AAT TGT GCT AAT CAG ATA ACA TTC CCT GCC TTC TAG ATATGCGGTGGAGTAGTAGTATATGTTTGTTTCTATTCGAGAATTAGCAGTTTTCAATTTGTTTGCA AACTTGCAATTCAAGCCTAATAAACCGATTACTTTCATCAATTTGATTGAAAATAAAGTTTTACGGA TGTATTAAGCATTTTGTAATCTGATTATGGATTTATTGTGTCTGTGTATGCTACTTATTCTCTAGAA GATGGTATTTGACACTAAAAAGTAATCATTGTATTCAAAATTCTATCTACTTAGATAATATTTAAAA

References

Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE)

database: 1999. Nucleic Acids Res 27:297-300

Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S (2002)

PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis

of promoter sequences. Nucleic Acids Res 30:325-327

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Fig. S2 DNA methylation profiles in and around promoters. For each methylation context, filled and

open circles indicate a methylated and unmethylated status at cytosine residues in CG, CHG and CHH (H

= A, C, or T), respectively. Transcription start site is indicated with “+1”. (a) Profile for da-I for DNAs

from glandular tentacles, laminas, roots, and inflorescences. (b) Profile for dm-I for DNAs from secretory

glands, marginal bristles, laminas, roots, and inflorescences. (c) Profile for cf-I for DNAs from secretory

glands, pitchers with secretory glands removed, normal leaves, and rhizomes and adventitious roots.

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Fig. S3 Phylogenetic relationship among the plants of interest. A part of the angiosperm phylogeny

reported by Stevens (2001) is shown along with the plant names listed in Table 1. Names of RNases are

given in parentheses.

Reference

Stevens PF (2001 onwards) Angiosperm Phylogeny Website. Version 12, July 2012.

http://www.mobot.org/MOBOT/research/APweb. Accessed 25 January 2013

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