0.98
0.4
1
0.54
0.28
0.35
0.33
1
0.58
0.32
0.97
0.4
0.54
1
0.88
0.47
1
1
0.4
0.31
0.44
0.28
0.89
0.33
0.45
0.96
0.95
1
1
1
0.98
0.88
1
0.47
0.93
0.48
1
0.28
0.7
1
1
0.53
0.32
0.98
1
0.99
Nt Caffeic acid-OMT (AAL91506.1): CafCoA
Mc lAOMT (M87340): myoInositol
Ms IlOMT (AAB48059.1): isoliquiritigenin2
Lp Caffeic acid OMT (AAD10253.1): Caffeic acid
Ta
flavone (tricetin)
B2
Tt
catechol
Ca
methylated flavonols
At OMT1-At5g54160 (AAB96879.1)-flavonols
Ms
CafCoA
Sl OMT3 (AK325603): unknown
Sc OMT (AAO23335.1): unknown
Cr OMT2 (AAM97497.1): myricitin & dihydromyricitin
Cr OMT4-(AAR02419.1): unknown
Mp
flavones
B1
Vv ROMT (CAQ76879.1): resveratol
Rh
Lj
Ms
isoflavone
Ge
Zv
unknown
Nt
CafCoA
At 4g34050 (AAL09793.1): unknown
Ms
CafCoA
Nt
CafCoA
Vv
CafCoA
Pt
unknown
Pt
unknown
Pc
CafCoA
Os 06g0165800 (NP001056910.1): unknown
Zm
unknown
Zm
unknown
A1
Zm
unkown
Os 08g0498100 (NP001062142.1): unknown
At 1g67980 (NP564916.2): unknown
Vv AOMT3 (CBI27641.3): unknown
Vv AOMT1 (ACO52469.1)-Anthocyanin
Vv AOMT2 (HQ702997): unknown
Ph DIFe1 (MT2)-Anthocyanin
St
unknown
Ph DIFe2a (MF1)-Anthocyanin
Ph DIFe2b (MF2)-Anthocyanin
Ckm AOMT2 (AB618483) unknown
Ckm AOMT1 (AB618482.1): anthocyanin ?
At
CafCoA, Flavonols, Caf-esters
Mc
CafCoA, Flavonols, Caf-esters
Stl
CafCoA, Flavonoids, Caf-esters
Am
Benzoic Acid
A2
At
Jasmonic Acid
Cb
Salicylic Acid
Supplemental Figure 1. Cladogram of plant O-methyltransferases.
The tree (neighbourjoining, BioNJ) is based on the alignment of a selection of full protein sequences of
methyltransferases (cf. Supplemental Data Set 2) belonging to the A1, A2, B1 and B2 subfamilies (Lam et
al., 2007). Red numbers indicate bootstrap support for each branch (1000 replicates). For each protein the
name, accession number, and substrates accepted in vitro or in vivo is given. Prefixes indicate the species of
origin: Am, Antirrhinum majus; At, Arabidopsis thaliana; Ca, Chrysosplenium americanum; Cb, Clarckia
breweri; Cr, Catharantus roseus; Ckm, Cyclamen persicum x purpurascens; Ge, Glycyrrhiza echinata; Lj,
Lotus japonicus; Lp, Lolium perenne; Mc Mesembryanthemum crystallinum; Mp, Mentha piperita; Ms,
Medicago sativa; Nt, Nicotiana tabacum; Os, Oryza sativa; Pc, Petroselinum crispum; Ph, Petunia hybrida;
Pt, Populus trichocarpa; Rh, Rosa hybrid; Sc, Secale cereale; Sl, Solanum lycopersicum; St, Solanum
tuberosum; Stl, Stellaria longipes; Ta, Triticum aestivum; Tb, Thalictrum tuberosum; Vv Vitis vinifera; Ze,
Zinnia elegans; Zm, Zea mays; Zv, Zinnia violacea.
1
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
ATGACAGGCAAAACTGCCCACCCTGGCATTCTACGAAGTGATGCCCTCAGGAAGgttaatatttactgcttgttttattcatcttcattacttaatttttaaccgcacttgc
ATGACAGGCAAAACCGCCCACCCTGGCATTCTTCGAAGTGATGCCCTCAGGAAGgttaatatttactgcttgttttattcatcttcattacttaattcttaaccgcacttgc
ATGACAGGCAAAACCGCCCACCCTGGCATTCTTCGAAGTGATGCCCTCAGGAAGgttaatatttactgcttgttttattcatcttcattacttaatttttaaccgcacttgc
ATGACAGGCAAAACCGCCCACCCTGGCATTCTTCGAAGTGATGCCCTCAGGAAGgttaatatttactgcttgttttattcatcttcattacttaatttttaaccgcacttgc
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
tcaacaagatatataatacttacttatatatgtacttaaagtcttaaacaaaagctaatttcggatgcattatgcatgtgtaattataaaacaactatttttaccttggacg
tcaacaagatatttag----tacttatatatgtactttaagtcttaaacaaaagttaatttcggatgcattatgcatgtgaaattat-aaacaactctttttaccttggaca
tcaacaagatatttag----tacttatatatgtactttaagtcttaaacaaaagttaatttcggatgcattatgcatgtgaaattat-aaacaactctttttaccttggaca
tcaacaagatatttag----tacttatatatgtactttaagtcttaaacaaaagttaatttcggatgcattatgcatgtgaaattat-aaacaactctttttaccttggaca
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
ttaaaatgaactctactctgtttcaattgaatattgattaacttgatactaa----tttagtttatattagctatctgcccatagtattgtcataactaaatgtgagctata
ttataatgaactctactctgtttcaattgaacattgattaacttgctaccac--tacttttagttcattagctatctgcccataatattgtcaaaactaaatgtgggctata
ttataatgaactctactctgtttcaattgaacattgattaacttgctaccac--tacttttagttcattagctatctgcccataatattgtcaaaactaaatgtgggctata
ttataatgaactctactctgtttcaattgaacattgattaacttgctaccac--tacttttagttcattagctatctgcccataatattgtcaaaactaaatgtgggctata
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
tacgcacacaaaaaatatcaatagtaaacagcaaatggggtggactaatctgcttcaacttgttttctcttttattttattaca-gTACATTTTGGAAACCTCAGTTTATCC
tacgcacactaaaaatatcaacagtaaagagcaaatggggtggactaatttgcttcaacttgttttctcttttattttattaca-gTACATTTTGGAAACCTCAGTTTATCC
tacgcacactaaaaatatcaacagtaaagagcaaatggggtggactaatttgcttcaacttgttttctcttttattttattaca-gTACATTTTGGAAACCTCAGTTTATCC
tacgcacactaaaaatatcaacagtaaagagcaaatggggtggactaatttgcttcaacttgttttctcttttattttattacaggTACATTTTGGAAACCTCAGTTTATCC
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
AAGAGAGCATGAGCTACTCAAAGAACTCACAAAAGCTTCATTCGAAAAGTATAAAGCAGCgtaagtccctcctacacccatattctgacctatattttggattaact----AAGAGAGCATGAGCTACTCAAAGAACTAACAAAAGCTTCATTCGAGAATTATAAAGCAGCgtaagt-cgtcctacacccatatgctgacctatattttggattaacttatac
AAGAGAGCATGAGCTACTCAAAGAACTAACAAAAGCTTCATTCGAGAATTATAAAGCAGCgtaagt-cgtcctacacccatatgctgacctatattttggattaacttatac
AAGAGAGCATGAGCTACTCAAAGAACTAACAAAAGCTTCATTCGAGAATTATAAAGCAGCgtaagt-cgtcctacacccatatgctgacctatattttggattaacttatac
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
---tataaacaaacagtgtgaaacctttttacagtaatacatttaacctcgtatagttatatattttgccttatcttttatgtt--attgt------tctcattttttgtgg
ttatatacacaaacagtgtaacacctttt---agtaacatatttaacctcttgtagcta-ataacttgtcgtatttctcatatttaattgtttatgatctcactttttatgg
ttatatacacaaacagtgtaacacctttt---agtaacatatttaacctcttgtagcta-ataacttgtcgtatttctcatatttaattgtttatgatctcactttttatgg
ttatatacacaaacagtgtaacacctttt---agtaacatatttaacctcttgtagcta-ataacttgtcgtatttctcatatttaattgtttatgatctcactttttatgg
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
acagttatctatagttattttttagctaggtgacctgattattatacattgtcggtaaatacaagtaatttcaaatactcttggcatgtac-atgcacaagttttacctatt
aca-------acagttatctt-----taggtgacttgat------------------aatgtgagaattgcttaacattattagtatatactacttagaagttgaagtta-aca-------acagttatctt-----taggtgacttgat------------------aatgtgagaattgcttaacattattagtatatactacttagaagttgaagtta-aca-------acagttatctt-----taggtgacttgat------------------aatgtgagaattgcttaacattattagtatatactacttagaagttgaagtta--
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
acccaattagtagtat-------atagtagaagttaaactcgtactat-atcacctaaattgc------------------atcaacttatacttgtcttaacattttaagt
----aactggtaatgtcgtagccacagtgggcaattgcccctgcctatcgtagtttaaattgcatcgatgttaaaactcaaatcaaattatacttgtcttaacattttaact
----aactggtaatgtcgtagccacagtgggcaattgcccctgcctatcgtagtttaaattgcatcgatgttaaaactcaaatcaaattatacttgtcttaacattttaact
----aactggtaatgtcgtagccacagtgggcaattgcccctgcctatcgtagtttaaattgcatcgatgttaaaactcaaatcaaattatacttgtcttaacattttaact
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
gtttattattatttcgtaaatatatctatctatacagGAGCTTTATGGGTCTTCCTCAAGATGAAGCCCAATTCCTATCGATGTTCCTAAAGCTCATAAATGCAAAGAAAAC
gtttactattatttcg------tatatatttatacagGAGCTTTATGGGTCTTCCTCAAGATGAAGCCCAATTCCTATCGATGTTCCTAAAGCTCATAAATGCAAAGAAAAC
gtttactattatttcg------tatatatttatacagGAGCTTTATGGGTCTTCCTCAAGATGAAGCCCAATTCCTATCGATGTTCCTAAAGCTCATAAATGCAAAGAAAAC
gtttactattatttcg------tatatatttatacagGAGCTTTATGGGTCTTCCTCAAGATGAAGCCCAATTCCTATCGATGTTCCTAAAGCTCATAAATGCAAAGAAAAC
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
ACTAGAGATTGGAGTATTCACTGGTTACTCTCTGCTTGTTACTGCTCTTGCTTTGCCAGAAGATGGTAAAgtaagtcaaataaacatatcacaatcatatgatcttattaaa
ACTAGAGATTGGAGTATTCACTGGTTACTCTCTGCTTGTTACTGCTCTTGCTTTGCCAGAAGATGGTAAAgtaagtctaataaa-atatcacaatcatatgatcttattaaa
ACTAGAGATTGGAGTATTCACTGGTTACTCTCTGCTTGTTACTGCTCTTGCTTTGCCAGAAGATGGTAAAgtaagtctaataaa-atatcacaatcatatgatcttattaaa
ACTAGAGATTGGAGTATTCACTGGTTACTCTCTGCTTGTTACTGCTCTTGCTTTGCCAGAAGATGGTAAAgtaagtctaataaa-atatcacaatcatatgatcttattaaa
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
cagagttttcttgaaaatgtcccttagatgttaatatgtaattatccattgat-caatgtacagGTAATAGCAATTGACCCGGACAGAGAGGCATATGAGGTTGGATTACCT
cagagttttcttgaaaatgtcccttagatgttaatatgtaattatccattgat-caatgtacagGTAATAGCAATTGACCCGGACAGAGAGGCATATGAGGTTGGATTACCT
cagagttttcttgaaaatgtcccttagatgttaatatgtaattatccattgat-caatgtacagGTAATAGCAATTGACCCGGACAGAGAGGCATATGAGGTTGGATTACCT
cagagttttcttgaaaatgtcccttagatgttaatatgtaattatccattgatccaatgtacagGTAATAGCAATTGACCCGGACAGAGAGGCATATGAGGTTGGATTACCT
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
TATATTCAGAAGGCTGGTGTGGAACATAAGATCGAGTTCATTCAATCAGATGCCGTGCCCGTTCTTGAAAAACTCCTCTTTAACgtatggcttcatccttcgtaggagtaaa
TATATTCAGAAGGCTGGTGTGGAACATAAGATCGAGTTCATTCAATCAGAAGCCGTGCCCGTTCTTGAAAAACTCCTCTCTAACgtatggcttcatccttcgtaggagtaaa
TATATTCAGAAGGCTGGTGTGGAACATAAGATCGAGTTCATTCAATCAGAAGCCGTGCCCGTTCTTGAAAAACTCCTCTCTAACgtatggcttcatccttcgtaggagtaaa
TATATTCAGAAGGCTGGTGTGGAACATAAGATCGAGTTCATTCAATCAGAAGCCGTGCCCGTTCTTGAAAAACTCCTCTCTAACgtatggcttcatccttcgtaggagtaaa
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
tatttttttatactatcagtgtattgt--aaca-----------------------------tattataaatgattccattttttaagttacggtagactttctttttgcta
ta-ttttttatactatcaatgtattgtacaacatattataaa<2x dTph11(~2400Bp)>tattataaatgattctattttttaagttacggtagacttt-tttttgtta
ta-ttttttatactatcaatgtattgtacaaca-----------------------------tattataaatgattctattttttaagttacggtagacttt-tttttgtta
tatttttttatactatcaatgtattgtacaaca-----------------------------tattataaatgattctattttttaagttacggtagacttt-tttttgtta
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
taggtaacttaatctaatgatataaaataattttatgctaatgttttcccatttaattgaaatactatactctattatatatgttaatgttgatattcaaaatgatatcagG
taggtaacttaatctaatgatataaaataattttatactaatgttttcccatttaattgaaatactatactctat--tatatgttaatgttgatattcaaaatgacatcagG
taggtaacttaatctaatgatataaaataattttatactaatgttttcccatttaattgaaatactatactctat--tatatgttaatgttgatattcaaaatgacatcagG
taggtaacttaatctaatgatataaaataattttatactaatgttttcccatttaattgaaatactatactctat--tatatgttaatgttgatattcaaaatgacatcagG
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
AGAAAGAAGCAGGGACATTTGATTTCGTGTTCATTGATGCTGATAAGGAGAATTATTTGAAGTACCATGAGATAGTGCTAAAATTGGTGAAAGTTGGAGGAGTGATAGGATA
AGAAAGAAGCAGGGACATTTGATTTCGTGTTCATTGATGCTGATAAGGAGAATTATTTGAAGTACCATGAGATAGTGCTAAAATTGGTGAAAGTTGGAGGAGTGATAGGATA
AGAAAGAAGCAGGGACATTTGATTTCGTGTTCATTGATGCTGATAAGGAGAATTATTTGAAGTACCATGAGATAGTGCTAAAATTGGTGAAAGTTGGAGGAGTGATAGGATA
AGAAAGAAGCAGGGACATTTGATTTCGTGTTCATTGATGCTGATAAGGAGAATTATTTGAAGTACCATGAGATAGTGCTAAAATTGGTGAAAGTTGGAGGAGTGATAGGATA
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
TGACAACACCTTATGGTTTGGGACAGTGGCACTATCAGAGGATGATCCAATGCCAGAAGGTTTAAGAGCATTAAGGGGACATGTTATGAAGGTCAATAGCTTTTTAGCTACT
TGACAACACCTTATGGTTTGGGACAGTGGCACTTTCAGAGGATGATCCAATGCCAGAAGGTTTAAGAGCATTAAGGGGACATGTTATGAAGGTCAATAGCTTTTTAGCTACT
TGACAACACCTTATGGTTTGGGACAGTGGCACTTTCAGAGGATGATCCAATGCCAGAAGGTTTAAGAGCATTAAGGGGACATGTTATGAAGGTCAATAGCTTTTTAGCTACT
TGACAACACCTTATGGTTTGGGACAGTGGCACTTTCAGAGGATGATCCAATGCCAGAAGGTTTAAGAGCATTAAGGGGACATGTTATGAAGGTCAATAGCTTTTTAGCTACT
PiMT(S6)
PaMT(S26)
PaMT(S2)
PhMT(V30)
GACCCTCGTGTTGAAGTAGCTCAACTTTCAATTGGCGATGGCCTTACCCTTTGCAGACGTCTCTCCTAG
GACCCTCGTGTTGAAGTAGCTCAACTTTCAATTGGCGATGGCCTTACCCTTTGCAGACGTCTCTCCTAG
GACCCTCGTGTTGAAGTAGCTCAACTTTCAATTGGCGATGGCCTTACCCTTTGCAGACGTCTCTCCTAG
GACCCTCGTGTTGAAGTAGCTCAACTTTCAATTGGCGATGGCCTTACCCTTTGCAGACGTCTCTCCTAG
Supplemental Figure 2. Aligned coding sequences and introns of MT2 alleles. The coding sequence from
start to stop codon is depicted uppercase, intron sequences in grey lowercase letters. Dashes represent gaps
introduced to improve the alignment. The position of the double dTPH11 insertion and 8 –bp target site
duplication are shown in red.
2
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttc---ttacttaaatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttc---ttacttaaatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttcttattacttaaatgcatatcttgctctagcctt
ATGGCAGGCAAAGGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttc---ttacttaaatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttc---ttacttaaatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagttctcttc---ttactt-gatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagttctcttc---ttactt-gatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCAT--TCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagttctcttc---ttactt-gatgcatatcttgctctagcctt
ATGGCAGGCAAAAGCGGACATGGCTCCATGATCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagttctcttc---ttactt-gatgcatatcttgctctagcctt
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
actatgctcatgca---------------------------------------------------------------------------------------------------actatgctcatgca---------------------------------------------------------------------------------------------------actatgatcatgcagtgtgcatgtgtg--------------------------------------------------------------------------------------actatgctcatgcagtgtgcatgtgtgttatgaatgtggttgaaggagtattagttaatgggcctgtatataagagcaaatgtcaacatcagtccaaaacccgtttaagtgtct
actatgctcatgcagtgtgcatgtgtgttatgaatgtggttgaaggagtattagttaatgggcctgtatataagagcaaatgtcaacatcagtccaaaacccgtttaagtgtct
actctgttcatgaagtgtgcatgtg----------------------------------------------------------------------------------------actctgctcatgaagtgtgcatgtgtg--------------------------------------------------------------------------------------actctgctcatgaagtgtgcatgtgtg--------------------------------------------------------------------------------------actctgctcatgaagtgtgcatgtgtg---------------------------------------------------------------------------------------
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------atatttgggatccaggcccatctagcaatgagttaaaaagagttcacacaattttaacaagggattatgcatattgtgtatgatgccttaggcataatctatctttctcattcc
atatttgggatccaggcccatctagcaatgagttaaaaagagttcacacaattttaacaagggattatgcatattgtgtatgatgccttaggcataatctatc-ttctcattcc
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------tgtttcttctgattcttctcatctcccttctcccttatcgttactgctattacttcatcctattgtaatctgctcttttcaagtattaatcaaaggaacttgtttcattatttt
tgtttc-tctgat--ctctcatctcccttctcccttatcgttactgctattacttcatcctattgtaatctgctcttttcaagtattaatcaaaggaacttgtttcatta-ttt
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
------------------------------------gtgtgtcaatgtattgctcatcttttctatctatatggagtaaactat---tatctatatatagctgattttc----t
------------------------------------gtgtgtcaatgtattgctcatcttttctatctatatggagtaaactat---tatctatatatagctgattttc----t
---------------------------------tgtgtgtgtcaatgtattgctcatcttttctatctatatggagtaaaatattagtatctatatatagctgattttc----t
gtcgaagtgttaagagttcacttcaatgtgtcatgtgtgtgtcaatgtattgctcatcttttctatctatatggagtaaaatat---tatctatatatagctgattttc----t
gtcgaagtgttaagagttcacttcaatgtgtcatgtgtgtgtcaatgtattgctcatcttttctatctatatggagtaaaatat---tatctatatatagctgattttc----t
---------------------------------tgtgtgtgtcaatgcattgctcatcttttctatctatatggagtaaaatat---aatctat--atggctgattttc----t
---------------------------------tgtgtgtgtcaatgcattgctcatcttttctatctgtatggagtaaa-tat---tatctatacatagctgattttcttatt
---------------------------------tgtgtgtgtcaatgcattgctcatcttttctatctgtatggagtaaa-tat---tatctatacatagctgattttcttatt
---------------------------------tgtgtgtgtcaatgcattgctcatcttttctatctgtatggagtaaa-tat---tatctatacatagctgattttcttatt
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
ttcgtgtgttttccttacattttattgcagTATATCTTTGAAACTAGTGTTTATCCAAGAGAACACGAGCAACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAA--ttcgtgtgttttccttacattttattgcagTATATCTTTGAAACTAGTGTTTATCCAAGAGAACACGAGCAACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAA--ttcgtgtgttttccttacattttattgcagTATATCTTTGAAACTAGTGTTTATCCAAGAGAACACGAGCAACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAA--ttcgtgtgttttccttacattttattgcagTATATCTTTGAAACTAGTGTTTATCCAAGAGAACACGAGCAACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAA--ttcgtgtgttttccttacattttattgcagTATATCTTTGAAACTAGTGTTTATCCAAGAGAACACGAGCAACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAA--ttcgtgtgtttcttttacattttattgcagTATATCTTTGAAGCTAGTGTTTATCCAAGAGAACACGAGCAACTCAAAGAACTCACACAAGCCTTATTCGATAAGTATAAA--ttcgtgtgtttctcttacattttactgcagTATATCTTTGAAACTAGTGTTTATCCATGAGAACACGAGCAACTCAAAGATCTCACACAAGCCTCATTCGATAAGTATAAT--ttcgtgtgtttctcttacattttactgcagTATATCTTTGAAACTAGTGTTTATCCATGAGAACACGAGCAACTCAAAGATCTCACACAAGCCTCATTCGATAAGTATAAT--ttcgtgtgtttctcttacattttactgcagTATATCTTTGAAACTAGTGTTTATCCATGAGAACACGAGCAACTCAAAGATCTCACACAAGCCTCATTCGATAAGTATAAT---
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
-----------------ATAGTgtaagtcctgcaccacattgcacaacataacccaaagatccaacgtacgcatcattttattactgtaatcatttctatggttggctttta-t
-----------------ATAGTgtaagtcctgcaccacattgcacaacataacccaaagatccaacgtacgcatcattttattactgtaatcatttctatggttggctttta-t
-----------------ATAGTgtaagtcctgcaccacattgcacaacataacccaaagatccaacgtacacatcattttattactgtaataatttctatgat--gca-tcagt
-----------------ATAGTgtaagtcctgcaccacattgcacaacataacccaaagatccaacgtacccatcattttattactgtaatattttctatgat--gca-tcagt
-----------------ATAGTgtaagtcctgcaccacattgcacaacataacccaaagatccaacgtacccatcattttattactgtaatattttctatgat--gca-tcagt
-----------------ATAGTgtaagtcc----------tgcacaacataacccaaagatccaacgtacgcatcattttattacagtaatattttctatgat--gcattcagt
>7000bp insertionAGAGTgcaagtcc----------tgcacaacataacccaaagatccaacgtacgcatcactttattacagtaatattttctatgat--gcattcagt
>7000bp insertionAGAGTgcaagtcc----------tgcacaacataacccaaagatccaacgtacgcatcactttattacagtaatattttctatgat--gcattcagt
>7000bp insertionAGAGTgcaagtcc----------tgcacaacataacccaaagatccaacgtacgcatcactttattacagtaatattttctatgat--gcattcagt
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
tactgtaaagatcctttaatttttccaaactcatataatatcatacaaataatattttaagtgcattttgaatt--ctagcac-aacttattgtgggtatacatatacagGAGC
tactgtaaagatcctttaatttttccaaactcatataatatcatacaaataatattttaagtgcattttgaatt--ctagcac-aacttattgtgggtatacatatacagGAGC
tgcagtaaagatcctttaatttttccaaactcatg-aatatcatacaaataatattttaagtgcactttgaactagcgagcac-aacttattgtgggtatacatatacagGAGC
tgcagtaaagatcctttaatttttccaaactcatg-aatatcatacaaataatattttaagtgcactttgaa----ctagcactaacttattgtgggtatacatatatagGAGC
tgcagtaaagatcctttaatttttccaaactcatg-aatatcatacaaataatattttaagtgcactttgaa----ctagcactaacttattgtgggtatacatatatagGAGC
tgcagtaaagaccc-----ttttcccaaactcatg-tatatcatacaaataatattttaagtgcacttagaa----agtgcac-aacttattctgggtaaacatatacagGAGC
tgcagtaaagaccc-----ttttcccaaactcatg-aatatcatacaagtaatattttaagtgcattttgatt---ctagcac-aacttattctgggtatacatatacagGAGC
tgcagtaaagaccc-----ttttcccaaactcatg-aatatcatacaagtaatattttaagtgcattttgaatt--ctagcac-aacttattctgggtatacatatacagGAGC
tgcagtaaagaccc-----ttttcccaaactcatg-aatatcatacaagtaatattttaagtgcattttgaatt--ctagcac-aacttattctgggtatacatatacagGAGC
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGG-------------------TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGG-------------------TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTTTGGCTACTG
TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTTTGGCTACTG
TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTTTGGCTACTG
TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCGAAGAAGACAATGGAGATTAGAGTTTTTACTGGTTATTCTCTTCTGGCTACTG
TTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATGCTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTTTGGCTACTG
TTGATGGGTGTGCCTCCAGATGAAGCCCAGTTTCTCTCGATGCTCTTAAAAATAATGAATGCGAAGAAGACAATGGAGATTGGAGTTTTTACTGATTATTCTCTTCTGGCTACTG
TTGATGGGTGTGCCTCCAGATGAAGCCCAGTTTCTCTCGATGCTCTTAAAAATAATGAATGCGAAGAAGACAATGGAGATTGGAGTTTTTACTGATTATTCTCTTCTGGCTACTG
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
----------------------------------------------------------------------------------agacatcatatgctagagcgaattccacgta
----------------------------------------------------------------------------------agacatcatatgctagagcgaattccacgta
CTCTTGCATTGCCAGAAGAAGGAAAAgtgagtttgataaataaccataatcattacta----------------------ttaatcatcatatgctaga--aaatttcacgta
CTCTTGCATTGCCAGAAGATGGAAAAgtgagtttgataaataaccataatcatta-------------------------ttaatcatcatatgctagagaaaatttcacgta
CTCTTGCATTGCCAGAAGATGGAAAAgtgagtttgataaataaccataatcatta-------------------------ttaatcatcatatgctagagaaaatttcacgta
CTCTTGCATTGCCAGAAGATGGAAAAgtgagt---- -----------------------< 1352bp insertion>attaatcatcatatgctagagcgaatttcacgta
CTCTTGCATTGCCAGAAGATGGAAAAgtgaatttgataaataaccataatcattattaat< 1421bp insertion >ttaatcatcatatgctagagcgaatttcacgta
CTCTTGCATTGCCAGAAGATGGAAAAgtgaatttgataaataaccataatcattattaat< 1421bp insertion >ttaatcatcatatgctagagcgaatttcacgta
CTCTTGCATTGCCAGAAGATGGAAAAgtgaatttgataaataaccataatcattattaat< 1421bp insertion >ttaatcatcatatgctagagcgaatttcacgta
3
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
tgattaatgattgacttatttacttaaattaaagttattgtcatgtaag--aaaaagatgaaatgtatcctccttcctcgcaatt---------------------------tgattaatgattgacttatttacttaaattaaagttattgtcatgtaag-aaaaaagatgaaatgtatcctccttcctcgcaatt---------------------------tgattaatgattgaattatcaactttaattaaagttattgtcatataag--aaaaagatgaaatgtatcctc---cctcatcaaacaacaacatacccagtataatctcac-a
tgattaatgattgaattatcaactttaattaaagttattgtcatataag-aaaaaagatgaaatgtatcctccctcctcatcaaacaacaacatacccagtataatctcacaa
tgattaatgattgaattatcaactttaattaaagttattgtcatataagaaaaaaagatgaaatgtatcctccctcctcatcaaacaacaacatacccagtataatctcacaa
cggttaatgattgacttatctactttaattaaagttattgtcataaaagnaaaaaagatgaaatgtatcatccttcctcat-------------------------------cgattaacgattgacttatctactttaattaaagttattgtcatataag-agaaaagatgaaatgtatcctccttcctcat-------------------------------cgattaacgattgacttatctactttaattaaagttattgtcatataag-agaaaagatgaaatgtatcctccttcctcat-------------------------------cgattaacgattgacttatctactttaattaaagttattgtcatataag-agaaaagatgaaatgtatcctccttcctcat--------------------------------
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
-----------------------------------------------------------------------------aaagtactaaattttactcacta-tagcaataa-------------------------------------------------------------------------------aaagtactaaattttactcacta-tagcaataa--gtgaggtctggagagagtgagatgtacgcagaccataccccaccgaggtggagaggctatttccgatagaccctcagaaagtactaaattttacttacta-taacaatagttg
gtggggtctggagagagtgagatgtacgcagaccataccccaccgaggtggagaggctatttccgatagaccctcgaaaagtactaaattttactcacta-taacaataa--atggggtctggagagagtgagatgtacgcagaccataccccaccgaggtggagaggctatttccgatagaccctcgaaaagtact-aattttactcacta-taacaataa-------------------------------------------------------------------------------aaagtactaaattttactcactattaacaatag-------------------------------------------------------------------------------aaagtactaaattttactcacta-taacaatag-------------------------------------------------------------------------------aaagtactaaattttactcacta-taacaatag-------------------------------------------------------------------------------aaagtactaaattttactcacta-taacaatag---
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
-----aattacccaaaaaaaaaatctatcattttaatgtaaactttgctcatataagggtaaagtcgctagctttacccactataatga-aaagttaaaactttaccatca------aattaccc-aaaaaaaaatctatcattttaatgt-aactttgctcatataagggtaaagtcgctagcttacccactataatga-aaagttaaaactttaccatca--t
atgaagattgct----------atttctcaggtatgtatgatt-----------------------ttggatctttctgcct-------------taggtcc-----ttct------aattaccc---aaaaatatctatcatttaattttaaat-----------------------ataaactttgctcattataagggtaaagcca--gcttcaccatca------aattaccc---aaaaatatctatcatttaattttaaat-----------------------ataaactttgctcattataagggtaaagcca--gcttcaccatca------aattacc-------atttttttttttgtcatttt-aac-----------------------gtaaactttacccactataatg--aaagttaaaacctcaccatcaac
-----aattacc--aaaaaaaaatttgttttgtcatttt-aac-----------------------gtaaactttacccactataatga-aaagttaaagcctcaccatca------aattacc--aaaaaaaaatttgttttgtcatttt-aac-----------------------gtaaactttacccactataatga-aaagttaaagcctcaccatca------aattacc--aaaaaaaaatttgttttgtcatttt-aac-----------------------gtaaactttacccactataatga-aaagttaaagcctcaccatca--
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
-----aagttgtc-----atatatcatactccctcc---------------------------ggtccat---tttagtgga--------------taggtttgcttttcgag
-----aag-tgtc-----atatatcatactccctcc---------------------------ggtccat---tttagtgga--------------taggtttgcttttcgag
----aggtatgtgctatcacttgtcaaagatcatct----caagctgattaacttttctatagaatcaaatggttgaattggaaaccataaacgggtacttatgaaaccctgt
-----aggttgtcttactatatatcatagtatatttttgttatactgaataat----------gctcaattactttaatgtgaga-aataa-----tagttttgtaact---t
----agggttgtcttactatatatcatagtatatttttgttatactgaataat----------gctcaattactttaatgtgaga-aataa-----tagttttgtaact---t
tatcaaggttgccttactatatatcatagtatattagtg-tatactgaataat----------gctcaattactttaatgtgaga-aataa-----tagttttgttacttact
-----aggttgccttactatatagcatagtatattagtg-tatactgaataat----------gctcaattactttaatgggaga-aataa-----tagttttgtaact---t
-----aggttgccttactatatagcatagtatattagtg-tatactgaataat----------gctcaattactttaatgggaga-aataa-----tagttttgtaact---t
---- aggttgccttactatatagcatagtatattagtg-tatactgaataat----------gctcaattactttaatgggaga-aataa-----tagttttgtaact---t
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
agtcaagttgaccaatcttcggagttaaattgaactagatctattcaatattataa--aattataattta-----ga--------tcgatattaaaaaattacacgaaaagta
agtcaagttgaccaatcttcggagttaaattgaactagatctattcaatattataa--aattataattta-----ga--------tcgatattaaaaaattacacgaaaagta
-gtctggcctgctttctctaagcgctgcgcctgt------tcgtttggaattgctatttctcttcttcttttccctattttttatttactatttcaaaagtttgtcattt--agtatagt------------agaataaagttcaa------ttatttaacgcaacaagtaattttttcttttt---ga------acttacctttatagtgatacgtgaaa---agtatagt------------agaataaagttcaa------ttatttaacgcaacaagtaattttttcttttt---ga------acttacctttatagtgatacgtgaaa---agtatagt------------agaataaattttaa------ttatttaacgcaataagtagttttttttttttttgga------acttacttttatagtgatacgtgaaattag
agtatagt------------agaataaagttcaa------ttatttaacgcaataagtagttttttttttttt--ga------acttactttcatagtgatacgtaaaattaa
agtatagt------------agaataaagttcaa------ttatttaacgcaataagtagttttttttttttt--ga------acttactttcatagtgatacgtaaaattaa
agtatagt------------agaataaagttcaa------ttatttaacgcaataagtagttttttttttttt--ga------acttactttcatagtgatacgtaaaattaa
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
ctacaagttagcaattcttttcatat-cactaag---cgtgaaaaggac----tcaaa---------------ttagtagaatcatttctat--------------------t
ctacaagttagcaattcttttcatat-cactaag---cgtgaaaaggac----tcaaa---------------ttagtagaatcatttctat--------------------t
tta------aact-------tgagaaggac---attttggtaaaatgactttttcaaaacaagggttcatgcttttat--agtaatatatatatatatatgatgatatgatat
cta------agcg-------tcatatgcactaagtttcgtgaaaaggac----tcaaa---------------ttagt--aatagtttctat--------------------t
cta------agcg-------tcatatgcactaagtttcgtgaaaaggac----tcaaa---------------ttagt--aatagtttctat--------------------t
cta------agca-------tcatatgcactaagtttcgtgaaaaagcc----tcaaa---------------ttagt--aatcgtttctat--------------------t
cta------agca-------tcatatgcactaagtttcgtgaaaaagac----tcaaa---------------ttagt--aatcgtttccat--------------------t
cta------agca-------tcatatgcactaagtttcgtgaaaaagac----tcaaa---------------ttagt--aatcgtttccat--------------------t
cta------agca-------tcatatgcactaagtttcgtgaaaaagac----tcaaa---------------ttagt--aatcgtttccat--------------------t
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
taatggtacacagAATATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGAGTGGAGCATAAGATTGAATTTATTCAATCAGAAG
taatggtacacagAATATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGAGTGGAGCATAAGATTGAATTTATTCAATCAGAAG
gattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAG
tattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTGTTCAATCAGAAG
tattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGAGTGGAGCATAAGATTGAATTTATTCAATCAGAAG
aattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGTGTGGAGCAT----------------------AAG
aattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCCATATATTCAGAAGGCTGGTGTGGAGCAT----------------------AAG
aattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTTGATTGCCATATATTCAGAAGGCTGGTGTGGAGCAT----------------------AAG
aattggtacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTTGATTGCCATATATTCAGAAGGCTGGTGTGGAGCAT----------------------AAG
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
CCTTGCCAGTACTCGAAAAACTCCTCTCTAACgtatggtttcttgccttt-ttaatttcttgtaataatagtttctttatag--tgttacctacaaggaatagtcaaactat
CCTTGCCAGTACTCGAAAAACTCCTCTCTAACgtatggtttcttgccttt-ttaatttcttgtaataatagtttctttatag--tgttacctacaaggaatagtcaaactat
CCTTGCCAGTACTCCAAAAACTCCTCTCTAACgtatggtttcttgccttt-ttaatttcttgtaataatagtttctttatag--tgttacatacagggaatagtcaaactat
CCTTACCGGTGCTCGAAAAACTCCTCTCTAACgtatggtttctggccttt-------------------------------------------------------------CCTTACCAGTACTCCAAAAACTCCTCTCTAACgtatggtttcttgccctttttaatttcgtgtaataatagtttctttatagtatgttacctacagtgagtagttaaactat
CCTTACCAGTACTCGAAAAACTACTCTCTAACgtatggtttctggccttt-ttaatttcgtgtaataatagtttctttatagtatgttacctacagtgagtagttaaactat
CCTTACCGGTGCTCGAAAAACTCCTCTCTAACgtatggtttctggccttt-ttaatttcgtgtaataatagtttctttatagtatgttacctacagtgagtagttaaactat
CCTTACCAGTACTCGAAAAACTACTCTCTAACgtatggtttctggccttg-ttaatttcgtgtaataatagtttctttatagtatgttacctacagtgagtagttaaactat
CCTTACCAGTACTCGAAAAACTACTCTCTAACgtatggtttctggccttt-ttaatttcgtgtaataatagtttctttatagtatgttacctacagtgagtagttaaac
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
tcgccttaactgcac---tgtttcaaatatttataattattac-----tactaattatattgaacatttacaatgaaatgatcagGGGGAGGAAGAAGGAACATTCGAT
tcgccttaactgcac---tgtttcaaatatttataattattac-----tactaattatattgaacatttacaatgaaatgatcagGGGGAGGAAGAAGGAACATTCGAT
tcgccttaactgcaatattgtttcaaatatttataattattac-----tactaattatattgaacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTCGAT
-----ttaactgcaatattgtttcaaatatttataattattac-----tactaattatactgaacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTCGAT
tcgccttaactgcaa---tgtttcaaatatttataattattac-----tactaattatattgaacatttacaatgaaatgatcagGGTGATGAAGAAGGAACATTCGAT
tcgccttaactgcaa---tctttcaaatatttataattattac-----tactaattatactgaacatttacaatgaaatgatgagGGTGAGGAAGAAGGAACATTCGAT
tcgccttaactgcaa---tctttcaaatatttataattattactggagtactaattatactgaacatttacaatgaaatgatgagGGTGAGGAAGAAGGAACATTCGAT
tcgccttaactgcaa---tctttcaaatatttataattattac-----tactaattatactgaacatttacaatgaaatgatgagGGTGAGGAAGAAGGAACATTCGAT
tcgccttaactgcaa---tctttcaaatatttataattattactggagtactaattatactgaacatttacaatgaaatgatgagGGTGAGGAAGAAGGAACATTCGAT
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTACCATGAGATAGTAC----TAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTACGACAACACATTATGG
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTACCATGAGATAGTAC----TAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTACGACAACACATTATGG
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTACCATGAGATAGTAC----TAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTATGACAACACATTATGG
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTACCATGAGATAGTAC----TAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTATGACAACACATTATGG
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTACCATGAGATAGTAC----TAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTACGACAACACATTATGG
TTCATATTCATTGATGCTGATACGGAGAACTATCTGAAGTACCATGAGATAGTACTAATTAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTATGACAACACATTATGG
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTAC------------------------------------------------------------------TTCATATTCATTGATGCTGATACGGAGAACTATCTGAAGTACCATGAGATAGTACTAATTAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTATGACAACACATTATGG
TTCATATTCATTGATGCTGATAAGGAGAACTATCTGAAGTACCATGAGATAGTAC----AAAAATTGGTGAAAGTTGGAGGAGTGATAGGCTATGACAACACATTATGG
4
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
TTTGGGACTGTGGCACTTTCAGATGATGATCCTATACCACAAGGCTTACGAGAATTGAGGAGATCTGTTTTGAAGATCAACAGCTTCTTAGCCACTGATCCTCGTATTG
TTTGGGACTGTGGCACTTTCAGATGATGATCCTATACCACAAGGCTTACGAGAATTGAGGAGATCTGTTTTGAAGATCAACAGCTTCTTAGCCACTGATCCTCGTATTG
TTTGGGACTGTGGCACTTTCAGATGATGATCCTATACCAGATGGCTTAAGAGAATTGAGGAGATCTGTTTTGAAGATCAACAGTTTCTTAGCTACTGATCCTCGCATTG
TTTGGGACTGTGGCACTTTCAGATGATGATCCTATACCAGATGGCTTAAGAGAATTGAGGAGATCTGTTTTGAAGATCAACAGTTTCTTAGCCACTGATCCTCGCATTG
TTTGGGACTGTGGCACTTTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCTGTTTTGAAGATCAACAGTTTCTTAGCTACTGATTCTCGCATTG
----------------------------------------------------------------------------------------------------------------------------TTTCAGATGATGATCCTATACTGGAAGGCTTAAGAGAATTGAGGAGA----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
Ph
Ph
Ph
Pi
Ph
Ph
Pa
Pa
Pa
mf1(R100)
mf1(R78)
MF1(V23)
MF1(S6)
MF1(M1)
mf1(V30)
mf1(S1)
mf1(S2)
mf1(S26)
AATTAGCTCATCTTTCAATTGGTGATGGCCTTACCCTCGGCAGGCGTCTCAATTAG
AATTAGCTCATCTTTCAATTGGTGATGGCCTTACCCTCGGCAGGCGTCTCAGCTAG
AATTAGCTCATCTTTCAATTGGTGATGGTCTAACCCTTGGCAGGCGTCTCAGCTAG
AATTAGCTCATCTTTCAATTGGTGATGGTCTTACTCTTGGCAGGCGTCTCAATTAA
AATTAGCTCATCTTTCAATTGGTGATGGTCTAACCCTTGGCAGGCGTCTCAATTAA
----------------------------------------CAGGCGTCTCAATT------------------------------------------------------------------------------------------------CAGGCGTCTCAATT---------------------------------------------------------
Supplemental Figure 3. Aligned coding sequences and introns of MF1 alleles. The coding sequence from
start to stop codon is depicted uppercase, intron sequences in grey lowercase letters. Deletions and insertions
that disrupt the coding sequence are highlighted in red and two large insertions in introns in blue.
5
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
ATGGCAGGCAAAAGCGGACATGGCTCCATTCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttcttatttaatgcatatctt
ATGGCAGGCAAAAGCGGACATGGCTCCATTCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagtttccttcttatttaatgcatatctt
ATGGCAGGCAAAAGCGGACATGGCTCCATTCTTCAAAGTGAAGCCCTCAAGAAGgttattagtttgaagagttcccttcttacttaatgcatatctt
ATGGCAGGCAAAAGCGGACATGGCTCCATTCTTCAAAGTGAAGCCCTCAAGAAGgttatt-------agagttcccttcttacttaatgcatatctt
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
gctctagcc----ttatttgctc
gctctagcc----ttatttgctc
gctctagccttaattacttgctc
gctctagccttaattacttgctc
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
atttta--tctatatagctgattttctc--gtgtgttctcttacattttattgcagTACATCTTCGAAACTAGTGTGTATCCAAGAGAACACGAGCA
atttta--tctatatagctgatttttctcgtgtgtttctcttacattttattgcagTACATCTTCGAAACTAGTGTGTATCCAAGAGAACACGAGCA
atctta--tctatgtagctgattttctt------tttctctcgcattttattacagTATATCTTCGAAACTAGTGTGTATCCACGAGAACACGAGCA
atattactcctatatagctgattttctt------tttctcttacattttattgcagTATATCTTCGAAACTAGTGTGTATCCAAGAGAACACGAGCA
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
ACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAAATAGTgtaagtcctgcacccgattgcactacataacccaaagatccaacatacacat
ACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAAATAGTgtaagtcctgcacccgattgcactacataacccaaagatccaacatacacat
ACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAAATAGTgtaagtcctgcacccgattgcaccacataacccaaatatccaacgtacacat
ACTCAAAGAACTCACACAAGCCTCATTCGATAAGTATAAAATAGTgtaagtcctgcacccgattgcactacataacccaaagatccaacgtacacat
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
cattttattactgttatattttatatatatgatgcatcaattgcaataaagatccttttcccaaacgagta---aaatgcttaaaaggtcactcaac
cattttattactgttatattttatatatatgatgcatcaattgcaataaagatccttttcccaaacgagta---aaatgcttaaaaggtcactcaac
cattatattactgttatattt----tgtatgatgcatcagttgcagtaaagatcctttttccaaactcatacatgaatatcatacaaa----taaaa
catttaattactgttatattt----tatacgatgcatcagttacagtaaagatcctttaaccaaattcatacatgaatatcatacaaataattaaaa
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
ttttaagtgcactgtgaactaacacaacttattatgggcatacata-acagGAGCTTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATG
ttttaagtgcactgtgaactaacacaacttattatgggcatacatatacagGAGCTTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATG
ttttaagtgcactgtgaactaacacaacttattatgggcatacatatacagGAGCTTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATG
ttttaagtgcactgtgaactaacacaacttattatgggcatacatatacagGAGCTTGATGGGTGTGCCTCCAGATGAAGCCCAATTTCTCTCGATG
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
CTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTCTGGCTACTGCTCTTGCATTGCCAGAAGATGGAA
CTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTCTGGCTACTGCTCTTGCATTGCCAGAAGATGGAA
CTCTTAAAGATAATGAATGCAAAGAAGACAATGGAAATTGGAGTTTTTACCGGTTATTCTCTTCTGGCTACTGCTCTTGCATTGCCAGAAGATGGAA
CTCTTAAAAATAATGAATGCAAAGAAGACAATGGAGATTGGAGTTTTTACCGGTTATTCTCTTCTGGCTACTGCTCTTGCATTGCCAGAAGATGGAA
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
AAgtgagtttgataaataatattaatcattattaatgatcatagtccagagggaatttcccgtatgatcaatgattgacctatctactttaataaag
AAgtgagtttgataaataatattaatcattattaatgatcatagtccagagggaatttcccgtatgatcaatgattgacctatctactttaataaag
AAgtgagtttgataaataatcataatcattattaatgatcatattccagagtgaatttcccgtatgatcaatgaatgacttatctactttaataaag
AAgtgagtttgataaataatcataatcattattaatgatcatattccagagtgaatttcccgtatgatcaacgattgatttatctactttaataaag
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
ttattggcaattaaaaaactgaaatgtatcctccttcatcataaagtcactaaatattaactcactaacaataaaagttaccaaaaataagataat
ttattggcatataaaaaactgaaatgtatcctccttcatcataaagtcactaaatattaactcactaacaataaaagttaccaaaaataagataat
ttattgtcatataaaaaactgaaatgtatcctccttcattataaagtaactaaattttactcactaaaacaataaagttaccaaaaataaaataac
ttattgtcaattaaaaaactgaaatgtatcctc--tcatcat-aagtaactaaattttactc-ctaaaac-ataaagttaccaaaaataaaataac
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
ttatcatattaaatataatcttcgcatataagagtaaagttgttaactttacccactataatgaaaagttaaatttcgccatcaaggttgccgtac
ttatcatattaaatataatcttcgcatataagagtaaagttgttaactttacccactataatgaaaagttaaatttcgccatcaaggttgccgtac
ttatcatattaaacctaatcttcgcatat------------------------------------aagttaagcttcgccatcaaagttgccttac
ttatcatattaaacctaatcttcgcatataagt---------------------------------------------------------------
Ph
Pa
Pi
Ph
mf2(V23)
mf2(S26)
MF2(S6)
MF2(R100)
tatatatcatattatcattatataataatgttatacactgaacaatactcaattactttaatgtgacaaataatactagtttgtaac-tattactg
tatatatcatattatcattatataataatgttatacactgaacaatactcaattactttaatgtgacaaataatactagtttgtaacttattactg
tatataataatgttatactgaataatat----------------atactcaattactttaatgtgacaaataatagt--tttgtaacttattacta
tatatataatgttat---------------------actgaataatactcaattactttaatgttacaa--aataatagtttgtaacttattacta
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
mf2(R78)par
mf2(V30)par
MF2(S6)
MF2(R100)
tagaataaagttcaattattaacgcaacaaataaatacatttactt-tatagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
atagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
tagaataaagttcaattattaacgcaacaaataaatacatttacttttatagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
atagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
tatagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
tatagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
tagaataaatttcaattattaacgcaacaaatgaatatatttacttttatagtgatacgtgaaactaaccc-tcatatgcaccaagtttcctgaaa
tagaataaatttcaattattaacgcaacaaatgaatatatttac-tttatagtgatacgtgaaactaacccatcatatgcaccaagtttcctgaaa
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
MF2(V30)par
mf2(V30)par
MF2(S6)
MF2(R100)
aagatggactcaaat----tagtaatcatttctattaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aagatggactcaaat----tagtaatcatttctattaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aagatggactcaaat----tagtaatcatttctattaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aagatggactc-aat----tagtaatcatttctattaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aagatggactc-aat----tagtaatcatttctattaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aagatggactc-aat----tagtaatcatttctattaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aagactcaaat--------tagtaatcatttctgttaattgatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
aggactcaaat----tagctagtaatcgtttctattaattaatacacagATTATAGCGATTGATCCGGACAGAGAAGCATATGAGGTTGGATTGCC
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
MF2(V30)par
mf2(V30)par
MF2(S6)
MF2(R100)
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTC--CTCTAACgtatgctttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTC--CTCTAACgtatgctttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTC--CTCTAACgtatgctttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTC--CTCTAACgtatgctttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTCCTCTCTAACgtatgctttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTCCTCTCTAACgtatgctttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTCCTCTCTAACgtatggtttct
ATATATTCAGAAGGCTGGTGTGGAGCATAAGATTGAATTTATTCAATCAGAAGCCTTACCAGTACTCGAAAAACTCCTCTCTAACgtatggtttct
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
MF2(V30)par
mf2(V30)par
MF2(S6)
MF2(R100)
tgcccttttaatttcttgtta---taatagtttctttttagtgttacctacagggagtaatcaaactattcgcttaactgcaatttttcaaa---t
tgcccttttaatttcttgtta---taatagtttctttttagtgttacctacagggagtaatcaaactattcgcttaactgcaatttttcaaa---t
tgcccttttaatttcttgtta---taatagtttctttttagtgttacctacagggagtaatcaaactattcgcttaactgcaatttttcaaa---t
tgcccttttaatttcttgtta---taatagtttctttttagtgttacctacagggagtaatcaaactattcgcttaactgcaatttttcaaa---t
tgcccttttaatttcttgtta---taatagtttctttttagtgttacctacagggagtaatcaaactattcgcttaactgcaatttttcaaa---t
tgcccttttaatttcttgtta---taatagtttctttttagtgttacctacagggagtaatcaaactattcgcttaactgcaatttttcaaa---t
tgcctttttaatttcttgtta---taatagtttcttttttgtgttacctacagggagtagtcaaactattcgcttaactgcaatttttcaaatatt
tgcctttttaatttcttgttatagtaatagtttcttttttgtgttacctacagggagtagacaaactatttgcttaactgcaatttttcaaa---t
atgaagtgtgcgtgtgtgt----------------caatgtattactcagcttttctatctacatggagtaaa
atgaagtgtgcgtgtgtgt----------------caatgtattactcagcttttctatctacatggagtaaa
atgaagtatgcatgtatgtgtgcgtgtgtgtgcgtcaatgtattgctcaccttttctatctatatggagtaaa
atgaagtatgcatgtatgtgtgcgtgtgt------caatgtattgctcagcttttctatctatatggagtaaa
6
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
MF2(V30)par
mf2(V30)par
MF2(S6)
MF2(R100)
atttataattattactactaaaaatacgacttcaaatttttgat-atgattta---attagtatat-------aaggaatcacttaca--cagacg
atttataattattactactaaaaatacgacttcaaatttttgat-atgattta---attagtatataattgtcaaggaatcacttaca--cagacg
atttataattattactactaaaaatacgacttcaaatttttgat-atgattta---attagtatataattgtcaaggaatcacttaca--cagacg
atttataattattactactaaaaatacgacttcaaatttttgat-atgattta---attagtatataattgtcaaggaatcacttaca--cagacg
atttataattattactactaaaaatacgacttcaaatttttgat-atgattta---attagtatataattgtcaaggaatcacttaca--cagacg
atttataattattactactaaaaatacgacttcaaatttttgat-atgattta---attagtatataattgtcaaggaatcacttaca--cagacg
atttataattattactactaaaaataagacttcaaatttttgat-atgatttatttagtagtatgtaattgtcaaggaatcatttacacgcagacg
atttataattattactact-aaaataagacttcaaatttttgataatgattta---gttagtatgtcattgtcaaggaatcatttaca--cagatg
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
mf2(R78)par
mf2(V30)par
MF2(S6)
MF2(R100)
aaaattttctttgaaatactgtatttgtttggtcgttcactgctagtgacggtaaaagaaggcaccataaatagcaaaaatactcattctacgagc
aaaattttctttgaaatactgtatttgtttggtcgttcactgctagtgacggtaaaagaaggcaccataaatagcaaaaatactcattctacgagc
aaaattttctttgaaatactgtatttgtttggtcgttcactgctagtgacggtaaaagaaggcaccataaatagcaaaaatactcattctacgagc
aaaattttctttgaaatactgtatttgtttggtcgttcactgctagtgacggtaaaagaaggcaccataaatagcaaaaatactcattctacgagc
aaaattttctttgaaatactgtatttgtttggtcgttcactgctagtgacggtaaaagaaggcaccataaatagcaaaaatactcattctacgagc
aaaattttctttgaaatactgtatttgtttggtcgttcactgctagtgacggtaaaagaaggcaccataaatagcaaaaatactcattctacgagc
aaaattttctttgaaa-----tatttgtttggtcgttcactgctagccacggtaaaa-aagacaccataaataaagaaaatactcattctacgagc
-aaattttctttgaaa-----tatttgtgtggtcgttcactgctagccacgattaaaaaaggcaccataaatagcgaaaatactcgttctacgagc
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
mf2(R78)par
mf2(V30)par
MF2(S6)
MF2(R100)
agcgcgtcaggctaattatattcgacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agcgcgtcaggctaattatattcgacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agcgcgtcaggctaattatattcgacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agcgcgtcaggctaattatattcgacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agcgcgtcaggctaattatattcgacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agcgcgtcaggctaattatattcgacatttacaatgaaatgatcagGGTGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agtg--tcagcctaattatattgaacatttacaatgaaatgatcagGGAGAGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
agtgc--cagcctaattttattgaacatttacaataaaatgatcagGGCGGGGAAGAAGGAACATTTGATTTCATATTCATTGATGCTGATAAGGA
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
mf2(R78)par
mf2(V30)par
MF2(S6)
MF2(R100)
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACCGTGCCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACCGTGGCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACCGTGGCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACCGTGGCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACCGTGGCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACCGTGGCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACTGTGGCACT
GAACTATCTGAAGTACCATGAGATAGTACTAAAATTGGTGAAAGTGGGAGGAGTGATAGGCTATGACAACACATTATGGTTTGGGACTGTGGCACT
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
mf2(R78)par
mf2(V30)par
MF2(S6)
MF2(R100)
TTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCGGTTTTGAAGATCAACAGTTTTTTAGCTACTGATCCTCGCATTGAATT
TTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCGGTTTTGAAGATCAACAGTTTTTTAGCTACTGATCCTCGCATTGAATT
TTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCGGTTTTGAAGATCAACAGTTTTTTAGCTACTGATCCTCGCATTGAATT
TTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCGGTTTTGAAGATCAACAGTTTTTTAGCTACTGATCCTCGCATTGAATT
TTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCGGTTTTGAAGATCAACAGTTTTTTAGCTACTGATCCTCGCATTGAATT
TTCAGATGATGATCCTATACCACAAGGCTTAAGAGAATTGAGGAGATCGGTTTTGAAGATCAACAGTTTTTTAGCTACTGATCCTCGCATTGAATT
TTCAGATGATGATCCCATACCAGAAGGCTTAAGAGAATTGAGGAGATCTGTTTTGAAGATCAATAGTTTCTTAGCCACTGATCCTCGCATTGAATT
TTCAGATGATGATCCCATACCAGAAGGCTTAAGAGAATTGAGGAGATCTGTTTTGAAGATCAATAGTTTCTTAGCTACTGATCCTCGCATTGAATT
Ph
Pa
Pa
Pa
Ph
Ph
Pi
Ph
mf2(V23)
mf2(S2)par
mf2(S26)
mf2(S1)
mf2(R78)par
mf2(V30)par
MF2(S6)
MF2(R100)
AGCTCATCTTTCAATTGGTGATGGTCTTACCCTTGGCAGGCGTCTCAGCTAG
AGCTCATCTTTCAATTGGTGATGGTCTTACCCTTGGCAGGCGTCTCAGCTAG
AGCTCATCTTTCAATT-------------CCCTTGGCAGGCGTCTCAGCTAG
AGCTCATCTTTCAATT-------------CCCTTGGCAGGCGTCTCAGCTAG
AGCTCATCTTTCAATT-------------CCCTTGGCAGGCGTCTCAGCTAG
AGCTCATCTTTCAATT-------------CCCTTGGCAGGCGTCTCAGCTAG
AGCTCATCTTTCAATTGGTGATGGTCTTACCCTTGGGAGACGTCTCAATTAG
AGCTCATCTTTCAATTGGTGATGGTCTTACCCTTGGGAGACGTCTCAACTAG
Supplemental Figure 4. Aligned coding sequences and introns of MF2 alleles. The coding sequence from
start to stop codon is depicted uppercase, intron sequences in grey lowercase letters. Deletions that disrupt
the coding sequence are highlighted in red.
7
A
Anthocyanidin (520 nm)
uV(x105)
3.5
Anthocyanin (520nm)
uV(x106)
Cya
4.0
Cya-3-Rut
3.0
3.0
2.5
2.0
Peo
2.0
1.5
1.0
Peo-3-Rut
1.0
0.5 35S:MF2 (#5)
35S:MF (#5)
35S:MF2 (#7)
0.0
untransformed
3
4
5
6
7
8
9
10
11
5
Anthocyanidin (520 nm)
uV(x105)
B
35S:MF (#7)
untransformed
0.0
7.5
10
15
17.5
Anthocyanin (520nm)
uV(x106)
Cya
3.0
12.5
Cya-3-Rut
2.5
2.5
2.0
2.0
Del
1.5
Peo
1.5
1.0 35S:MT (#4)
1.0 35S:MT (#4)
35S:MT (#7)
35S:MT (#7)
0.5
2
3
0.5
4
5
6
7
8
9
11 min
10
Anthocyanidin (520 nm)
uV(x105)
C
2.5
Del
9
10
11
12
13
14
15
16
17
18
19
18
19
Anthocyanin (520nm)
uV(x106)
Peo-3-Rut
Cya-3-Rut
0.5
0.5 35S:VvOMT (#2)
0.0
35S:VvOMT (#5)
35S:VvOMT (#7)
2
D
8
1.0
Mal
1.0
0.0
7
1.5
Peo
2.0
6
2.0
Pet
1.5
Peo-3-Rut
3
35S:VvOMT (#2)
35S:VvOMT (#5)
35S:VvOMT (#7)
4
5
6
7
8
9
10
11
6
Anthocyanidin (520 nm)
uV(x105)
7
8
9
10
11
12
13
15
14
16
17
Anthocyanin (520nm)
uV(x106)
2.0
3.0
Del
2.5
Pet
1.5
2.0
1.5
1.0
Mal
1.0
Cya
0.5
0.5
35S:MF (#1)
untransformed
0.0
3
4
5
6
7
8
9
10
35S:MF (#1)
untransformed
0.0
5
11
7.5
10
12.5
15
17.5
Supplemental Figure 5. HPLC-PDA analysis of anthocyanidins and anthocyanins in petals expressing
35S:MT, 35S:MF2 or 35S:VvAOMT1.
(A) HPLC tracings of anthocyanins (right panel) and anthocyanidins (obtained by acid hydrolysis; left panel)
in R78 petal limbs (untransformed) and two siblings expressing 35S:MF2.
(B) HPLC tracings of anthocyanins and anthocyanidins in petal limbs of two 35S:MT2 transformants
(C) HPLC tracings of anthocyanins and anthocyanidins in petal limbs of two 35S:MT2 transformants.
35S:VvAOMT (#2 and #5) and a transformants lacking detectable transgene expression (#7).
(D) HPLC tracing of anthocyanins and anthocyanidins in R78xV32 petals and a transformant expressing
35S:MF2.
8
A
Inten x107
MS(+): P1
mAU (x10) 520nm
3.0
P1: Cya-3Rut
[M+-Rutinoside+H]+
Inten x107
MS/MS(+)
m/z 595.17
595.2
2.5
287.1
287.1
[M+-Rutinoside+H]+
1.0
[M+]+
[M+-Rhamnose+H]+
0.5
449.1
2.0
0.0
0.0
300
400
500
600
Inten. x107
MS(+): P2
1.0
P2: Peo-3Rut
700
m/z
100
[M+]+
0.0
B
10.0
12.5
15.0
[M+-Rhamnose+H]+
463.1
300
17.5 RT
(min.)
400
500
600
700
800
100
m/z
200
HO
O HO
[M -Rutinoside+H]
(287.1)
C
[M+-Rutinoside+H]+
(301.1)
O
P1: Cya-3Rut
[M -Rutinoside+H]
2.5
7.5
10.0
12.5
15.0
17.5 RT
(min.)
mAU (x10) 520nm
500
600
609.2
1.0
[M+-Rutinoside+H]+
700
800
m/z
[M+]+
0.0
400
500
600
700
800
m/z
+
[M -Rutinoside+H]
E
400
500
Inten x10
MS(+): P2
12.5
15.0
17.5 RT
(min.)
mAU (x10) 520nm
1.5
600
700
800
m/z
0.5
500
600
700
800
m/z
200
[M+-Rutinoside+H]+
300
100
200
300
[M+-Rhamnose+H]+
0.5
449.1
449.1
0.0
400
500
600
609.2
700
800
m/z
100
200
Inten x107
5.0 MS/MS (+)
m/z 609.19
+ +
[M ]
[M+-Rutinoside+H]+
300
301.1
12.5
15.0
17.5 RT
(min.)
400
m/z
[M+-Rutinoside+H]+
[M+-Rhamnose+H]+
301.1
10.0
m/z
[M+-Rutinoside+H]+
[M ]
287.1
400
287.1
2.5
7.5
m/z
301.1
+ +
Inten x107
MS(+): P2
0.0
400
[M+-Rutinoside+H]+
Inten x107
MS/MS (+)
1.0 m/z 595.17
595.2
[M+-Rutinoside+H]+
300
P1: Cya-3Rut
m/z
0.0
400
0.0
2.5
400
287.1
1.0
301.1
300
1.0
300
[M+]+
0.0
P2: Peo-3Rut
100
Inten x107
MS/MS (+)
2.0 m/z 609.2
609.2
[M+-Rutinoside+H]+
Inten x107
MS(+): P1
5.0
200
0.0
300
10.0
[M -Rutinoside+H]
0.5
7
7.5
m/z
+
463.1
100
1.0
[M+]+
+
287.1
0.5
301.1
400
+
[M+-Rhamnose+H]+
Inten x107
MS/MS (+)
m/z 595.17
595.2
0.0
0.0
300
0.0
300
1.0
100
200
Inten x106
MS/MS (+)
m/z 609.2
2.5
P1: Cya-3Rut
P2: Peo-3Rut
449.1
5.0
301.1
Inten x107
MS(+): P1
1.0
[M+-Rhamnose+H]+
0.0
2.5
2.0
[M+-Rutinoside+H]+
0.5
300
400
Inten x107
MS(+): P2
0.5
287.1
1.0
[M+]+
+
0.0
0.0
OH
Inten x10
MS/MS (+)
m/z 595.17
595.2
+
287.1
P2: Peo-3Rut
O
7
Inten x10
MS(+): P1
1.0
H 3C
HO
HO
7
OH
O
[M+-Rhamnose+H]+
(463.1)
O
OH
2.0
D
O HO
OH
O
mAU (x10) 520nm
3.0
OH
OH
O
HO
O
+
[M+-Rhamnose+H]+
(449.1)
OH
+
H 3C
HO
m/z
OH
+
OH
400
OCH 3
OH
+
300
Peonidin-3-rutinoside (609.2)
OH
O
m/z
0.0
Cyanidin-3-rutinoside (595.2)
HO
301.1
400
[M+-Rutinoside+H]+
2.5
301.1
7.5
300
5.0
[M+-Rutinoside+H]+
0.5
200
Inten. x107
MS/MS(+)
m/z 609.18
609.2
1.0
0.0
800
2.5
463.1
0.0
0.0
300
400
500
600
700
800
m/z
100
200
300
400
m/z
Supplemental Figure 6. LC-MS/MS(+) analysis of anthocyanins in R78 petals and siblings expressing
35S:MT2, 35S:MF2 or 35S:VvAOMT1..
(A) LC-MS/MS (+) analysis of control R78 petal limbs.
(B) Predicted structures and MS/MS(+) ion fragments for anthocyanins in peaks 1 and 2.
(C-E) LC-MS/MS (+) analysis of R78 petal limbs expressing 35S:MT (C), 35S:MF2 (D) or 35S:VvAOMT1
(E).
LC profiles (left panels) are identical to those in Figure 7 and shown for reference. Peaks 1 and 2 were
analyzed by MS(+) (middle panels), and ions indicated by [M+]+ by MS/MS(+) (right panels).
9
A
B
LC: R78xV32
mAU (x10) 520nm
Delphinidin-3-(p-coumaroyl)-rutinoside-5-glucoside
[C42H47O23]+ = 919.25
P1: m/z 919(303,465)
~Del-3RutCoum-5Glu
4.0
3.0
2.0
P2: m/z 919 (no MS/MS)
~Del-3RutCoum-5Glu
P3: m/z 935(303,465)
~Del-3RutCaf-5Glu
1 2
1 : [M+-Glc+H]+ = 757.2
2 : [M+-CoumRut+H]+ = 465.1
1.0
1 2 : [M+-Glc-CoumRut+H]+ = 303.0
0.0
10
15
20
RT (min.)
C
D
Inten x107
MS(+): P1
479.1
[M+]+
1.0
919.2
5.0
303.0
2.5
0.5
300
400
500
600
700
800
200
Delphinidin-3-(caffeoyl)-rutinoside-5-glucoside
[C42H47O24]+ = 935.25
0.5
300
400
500
600
919.2
465.1
303.0
0.0
300
400
500
600
700
800
900 m/z
F
MS(+): P3
Inten x106
Inten x106
[M+]+
487.1
2.5
935.2
295.2
7.5
5.0
441.3
1 2
MS/MS(+): m/z 935.25
[M+-Glc-CafRut+H]+
303.0
[M+-CafRut+H]+
465.1
2.5
0.0
1 : [M+-Glc+H]+ = 773.2
[M+]+
[M+-CoumRut+H]+
[M+-Glc
-CoumRut+H]+
900 m/z
MS(+): P2
479.1
Inten x107
1.0
0.0
0.0
E
MS/MS (+): m/z 919.2
303.0
465.1
Inten x106
0.0
300
400
500
600
700
800
900 m/z
200 300 400
500 600 700 800
900 m/z
2 : [M+-CafRut+H]+ = 465.1
1 2 : [M+-Glc-CafRut+H]+ = 303.0
Supplemental Figure 7. LC-MS/MS analysis of anthocyanins in R78xV32 petals.
(A) LC-profile of R78xV32 petal limbs with PDA detection at 520 nm.
(B) Possible structure of anthocyanin in peaks 1 and 2 and m/z ratios for ion (fragments) seen in MS/MS(+).
(C) MS/MS(+) spectra of ion fragments from anthocyanin in peak 1.
(D) MS(+) spectrum of peak 2. MS/MS(+) analysis of ion fragments was not possible due to low abundance
of this compound.
(E) Possible structure of anthocyanin in peak 3 and m/z ratios for ion (fragments) seen in MS/MS(+).
(F) MS/MS(+) analysis of anthocyanins in peak 3.
10
LC: R78xV32::35S:MT2
mAU (x10) 520nm
A
P4: Pet-3RutCoum-5Glu
2.0
P2: Del-3RutCoum-5Glu
P5: Pet-3RutCoum-5Glu
P1: Del-3RutCoum-5Glu
1.0
P3:Del-3RutCaf-5Glu
P6: Mal-3RutCoum-5Glu
0.0
10.0
B
C
MS(+): P1
Inten x107
12.5
15.0
17.5
20.0
RT (min.)
D
MS(+): P4
Inten x107
[M ]
1.0
479.1
MS(+): P6
[M+]+
486.1
1.0
919.3
Inten x106
323.1
5.0
+ +
933.3
[M+]+
2.5
0.0
500
750
m/z
250
MS/MS(+): m/z 919.3
Inten x106
303.0
[M+-CumRut+H]+
465.1
500
750
m/z
250
0.0
317.1
5.0
500
750
m/z
Malvidin-3-(p-coumaroyl)-rutinoside-5-glucoside
Inten x106
MS/MS(+): m/z 933.3
[M+-Glc-CoumRut+H]+ [M+-CoumRut+H]+
[M+-Glc-CoumRut+H]+
5.0
[C44H51O23]+ = 947.3
479.1
0.0
300
5.0
0.0
0.0
250
947.3
733.23
317.1
400
500
600
m/z
MS(+): P2
Inten x106
300
400
479.1
[M+]+
919.3
2.5
1.0
500
m/z
E Inten x10
MS(+): P5
Inten x107
486.1
[M+]+
0.0
500
750
5.0
0.0
m/z
Inten x106
MS/MS(+): m/z 919.3
[M+-Glc-CoumRut+H]+
303.0
[M+-CoumRut+H]+
465.1
500
750
m/z
MS/MS(+): m/z 933.3
Inten x106
317.1
400
500
m/z
Delphinidin-3-(p-coumaroyl)-rutinoside-5-glucoside
750
[M+-Glc-CumRut+H]+
[M+-CumRut+H]+
479.1
5.0
479.1
m/z
MS/MS(+): m/z 949.3
[M+-CoumRut+H]+
0.0
0.0
300
500
Inten x106
317.1
[M+-Glc-CoumRut+H]+
1.0
0.5
0.0
949.3
2.5
317.1
250
[M+]+
487.1
933.3
0.5
0.0
5.0
MS(+): P3
6
300
400
500
m/z
Petunidin-3-(p-coumaroyl)-rutinoside-5-glucoside
300
400
500
600
m/z
Petunidin-3-(p-caffeoyl)-rutinoside-5-glucoside
[C43H49O23]+ = 933.3
[C43H49O24]+ = 949.3
+
[C42H47O23] = 919.3
1 2
1 : [M -Glc+H]+ = 757.2
2 : [M+-CoumRut+H]+ = 465.1
1 2 : [M+-Glc-CoumRut+H]+ = 303.0
+
1 2
1 2
1 : [M+-Glc+H]+ = 771.2
2 : [M+-CoumRut+H]+ = 479.1
1 : [M+-Glc+H]+ = 787.2
2 : [M+-CafRut+H]+ = 479.1
1 2 : [M+-Glc-CoumRut+H]+ = 317.1
1 2 : [M+-Glc-CafRut+H]+ = 317.1
Supplemental Figure 8. LC-MS/MS(+) analysis of anthocyanins in R78xV32 petals expressing 35S:MT2
(A) LC-profile of R78xV32 petals expressing 35S:MT2 with PDA detection at 520 nm.
(B) MS(+) and MS/MS(+) spectra and putative structure of anthocyanins in peaks 1 and 2.
(C) MS(+) and MS/MS(+) spectra and putative structure of anthocyanins in peaks 4 and 5.
(D) MS(+) spectrum and putative structure of anthocyanin in peak 6. Due to low abundance of this
compound no MS/MS(+) spectra could be obtained
(E) MS(+) and MS/MS(+) spectra and putative structure of anthocyanin in peak 3.
11
A
D
mAU (x10) 520nm LC: R78xV32::35S:MF2
5.0
G
MS(+): P4
Inten x107
1.0
P4: Pet-3RutCoum-5Glu
[M+]+
P5: Pet-3RutCoum-5Glu
P1/2: Del-3RutCoum-5Glu
2.5
P6: Mal-3RutCoum-5Glu
0.0
[M+]+
486.1
2.5
479.1
0.5
317.1
P7: Mal-3RutCoum-5Glu
0.0
10.0
12.5
15.0
17.5
20.0
MS(+): P6
Inten x106
933.3
493.1
318.2 331.1
0.0
300
RT (min.)
947.3
400
500
600
700
800
300
900 m/z
400
500
600
700
800
900
m/z
B
MS(+): P2
Inten x106
919.25
+ +
2.5
[M ]
479.1
5.0
921.3
507.6
303.0520
Inten x106
400
500
600
700
800
900 m/z
331.1
479.1
E
MS/MS(+): m/z 919.2
303.05
[M+-Glc-CoumRut+H]+
Inten x10
[M+-Glc+H]+
[M+-CoumRut+H]+
2.5
465.1
600
700
800
900 m/z
MS(+): P5
400
1.0
500
6
600
700
800
900 m/z
MS(+): P7
Inten x10
3.0 282.2
697.2
[M+]+
502.1
713.2
947.3
1.0
757.2
0.0
300
H
2.0
302.2
[M+-Glc+H]+
785.2
0.0
933.3
486.1
[M+-CoumRut+H]+
493.1
315.1
771.2
500
6
2.0
5.0
400
[M+-Glc-CoumRut+H]+
2.5
[M+-Glc+H]+
[M+-CoumRut+H]+
300
MS/MS(+) m/z 947.3
Inten x106
0.0
300
0.0
300
C
MS/MS(+): m/z 933.3
[M+-Glc-CoumRut+H]+
5.0
2.5
0.0
7.5
Inten x106
317.1
7.5
400
500
600
700
800
900 m/z
Delphinidin-3-(p-coumaroyl)-rutinoside-5-glucoside
300
F
400
500
600
700
800
900 m/z
Petunidin-3-(p-coumaroyl)-rutinoside-5-glucoside
0.0
I
300
400
500
600
700
800
900
m/z
Malvidin-3-(p-coumaroyl)-rutinoside-5-glucoside
[C43H49O23]+ = 933.3
[C44H51O23]+ = 947.3
[C42H47O23]+ = 919.25
1 2
1 2
1 : [M+-Glc+H]+ = 757.2
2 : [M+-CoumRut+H]+ = 465.1
1 2 : [M+-Glc-CoumRut+H]+ = 303.0
1 2
1 : [M+-Glc+H]+ = 771.2
2 : [M+-CoumRut+H]+ = 479.1
+
+
+
1 2 : [M -Glc-CoumRut+H] = 317.1
1 : [M -Glc+H]+ = 785.2
2 : [M+-CoumRut+H]+ = 493.1
1 2 : [M+-Glc-CoumRut+H]+ = 331.1
Supplemental Figure 9. LC-MS/MS(+) analysis of anthocyanins in R78xV32 petals expressing 35S:MF2
(A) LC-profile of R78xV32 petals expressing 35S:MF2 with PDA detection at 520 nm.
(B) MS(+) and MS/MS(+) spectra of anthocyanin in peak 2.
(C) Possible structure of anthocyanin in peak 2 and ion (fragments) seen in MS/MS(+)
(D) MS(+) and MS/MS(+) spectra of anthocyanin in peak 4
(E) MS(+) spectrum of peak 5. MS/MS(+) analysis of ion fragments was not possible due to low abundance
of this compound.
(F) Possible structure of anthocyanin in peaks 4 and 5 and m/z ratios for ion (fragments) seen in MS/MS(+)
(G) MS(+) and MS/MS(+) spectra of anthocyanin in peak 6
(H) MS(+) spectrum of peak 7. MS/MS(+) analysis of ion fragments was not possible due to low abundance
of this compound
(I) Possible structure of anthocyanin in peaks 6 and 7 and m/z ratios for ion (fragments) seen in MS/MS(+)
12
A
mAU (x10) 520nm LC: R78xV32::35S:MF2
P4: Pet-3RutCoum-5Glu
5.0
P5: Pet-3RutCoum-5Glu
P1/2: Del-3RutCoum-5Glu
2.5
P6: Mal-3RutCoum-5Glu
0.0
P7: Mal-3RutCoum-5Glu
B
10.0
Inten x107
1.0
0.5
12.5
15.0
17.5
20.0
RT (min.)
Inten x105
MS(-): P4 (14.5)
MS/MS (-): m/z 931.2
769.2
949.2
5.0
465.1
931.2
0.0
400
500
C
600
700
800
900 m/z
623.14
315.04
477.1
2.5
0.0
300
400
500
600
700
800
900 m/z
Petunidin-3-(p-coumaroyl)-rutinoside-5-glucoside
[C43H49O23-2H-]- = 931.2
1 2
1 : [M+-Glc-2H-]- = 769.2
1 2 : [M+-Glc-CoumRut-2H-]- = 315.05
2 : [M+-CoumRut-2H-]- = 477.1
1 3 : [M+-Glc-CoumRut-2H-]- = 623.16
3
Supplemental Figure 10 LC-MS/MS(-) analysis of peak 4 from R78xV32 petals expressing 35S:MF2
(A) LC-profile of R78xV32 petals expressing 35S:MF2 (PDA detection at 520 nm).
(B) MS/MS(-) spectra of anthocyanin in peak 4
(C) Possible structure of anthocyanin in peak 4 and ion (fragments) seen in MS/MS(-)
13
mAU (x10) 520nm
A
Inten x108
P2: Del-3RutCoum-5Glu
P4: Pet-3RutCoum-5Glu
P5: Pet-3RutCoum-5Glu
1.0
0.5
1.0
MS(+) 639.2 (Mal-3Rut)
0.5
MS(+) 393.1 (Mal-3Glu)
0.0
13
Inten x10
14
15
16
17
18
MS(+): P1
7
19
20
Inten x10
1.0
14
15
16
17
18
19
20
RT (min.)
MS/MS(+): m/z 919.3
5.0
479.1
13
RT (min.)
6
303.0
[M+]+
2.0
919.3
[M+-Glc-CoumRut+H]+
465.1
Delphinidin-3-(p-coumaroyl)-rutinoside-5-glucoside
[M+-CumRut+H]+
[C42H47O23]+ = 919.3
2.5
0.0
0.0
500
750
m/z
Inten x10
[M+]+
1.0
300
400
500
479.1
1 2
1 : [M -Glc+H]+ = 757.2
2 : [M+-CoumRut+H]+ = 465.1
+
303.0
465.1
0.5
919.3
0.5
m/z
MS/MS(+): m/z 919.3
Inten x107
[M+-Glc-CoumRut+H]+
1.0
[M+-CoumRut+H]+
MS(+): P2
7
1 2 : [M+-Glc-CoumRut+H]+ = 303.0
0.0
0.0
MS(+): P4
Inten x107
[M+]+
m/z
MS/MS(+): m/z 933.3
Inten x106
486.1
500
400
300
m/z
750
500
C
MS(+)
1.5
P6: Mal-3RutCoum-5Glu
0.0
B
PDA
P1:~Del-3RutCoum-5Glu
P9a/b: Mal-3Rut /Mal-3Glu
1.5
317.1
933.3
479.1
[M+-Glc-CoumRut+H]+ [M+-CoumRut+H]+
5.0
5.0
Petunidin-3-(p-coumaroyl)-rutinoside-5-glucoside
[C43H49O23]+ = 933.3
2.5
0.0
0.0
250
750
500
m/z
MS(+): P5
Inten x107
300
400
317.1
1.0
5.0
[M+]+
472.1
479.1
0.5
m/z
1 2
[M+-Glc-CoumRut+H]+
[M+-CoumRut+H]+
1 : [M+-Glc+H]+ = 771.2
2 : [M+-ComRut+H]+ = 479.1
479.1
1 2 : [M+-Glc-CoumRut+H]+ = 317.1
933.3
0.0
0.0
500
D
500
MS/MS(+): m/z 933.3
Inten x106
Inten x10
750
300
m/z
500
400
323.9
5.0
m/z
Malvidin-3-(p-coumaroyl)-rutinoside-5-glucoside
MS(+): P6
6
[C44H51O23]+ = 947.3
[M+]+
495.1
2.5
947.3
0.0
500
E
750
m/z
MS(+): P9 (14.2 min)
Inten x107
493.1
Inten x107
331.1
639.2
Malvidin 3-rutinoside
MS/MS(+): m/z 639.2
+
[M -Rut+H]
[C29H35O16]+ = 639.2
+
1 : [M+-Rut+H]+ = 331.1
2 : [M+-Rha+H]+ = 493.1
[M+]+
331.1
1.0
[M+-Rha+H]+
493.1
0.0
250
500
m/z
250
m/z
500
1
2
MS(+): P9 (14.3 min)
Inten x107
[M+]+
1.0
493.1
Inten x107
1.0
639.2
Malvidin 3-glucoside
MS/MS(+): m/z 493.2
331.1
[M+-Rut+H]+
[C23H25O12]+ = 493.1
1
: [M+-Glu+H]+ = 331.1
0.5
0.5
0.0
250
500
750
m/z
0.0
200
300
400
500
m/z
1
Supplemental Figure 11. LC-MS/MS(+) analysis of anthocyanins in R78xV32 petals expressing
35S:VvAOMT1.
(A) LC-profile of R78xV32 petals expressing 35S:VvAOMT1 using PDA detection at 520 nm (left) or MS
detection of m/z 639.2 (delphinidin 3-rutinoside) and 393.1 (delphinidin 3-glucoside).
(B) MS/MS(+) spectra and putative structure of anthocyanins in peaks 1 and 2.
(C) MS/MS(+) spectra and putative structure of anthocyanins in peaks 4 and 4.
(D) MS/MS(+) spectra and putative structure of anthocyanin in peaks 6.
(E) MS/MS(+) spectra and putative structures of anthocyanins with retention times of 14.3 and 14.2 min in
peaks 9.
14
Supplemental Figure 12. Structural models of Ms CCoA-MT in complex with anthocyanin substrates.
(A) Crystal structure (PDB code: 1SUI) of Ms CCoA-MT showing residues close to the active site with
bound S-adenosyl-L-homocysteine (SAH, pink), Feruloyl-CoA product (brown) and the divalent cation cofactor (yellow) (Ferrer et al., 2005).
(B) Active site of Ms CCoA-MT in which Caffeoyl-CoA is replaced by delphinidin-3-glucoside (brown), in
a binding orientation that leads to steric clashes between the 3-glucose group and the conserved Trp193,
Asp238 and Asp190 residues.
(C) Side view of (B).
(D) Possible binding orientation of delphinidin 3-(coumaroyl) rutinoside.
(E) Top view of (D).
(F) Same model and viewpoint as in (E), except that a 5-glucose group (cherry red) is now added to
delphinidin 3-(coumaroyl)rutinoside. The 5-glucose group was docked to prevent steric overlap with Nterminal residues (comprising Lys21), which leads to clashes between the 5-glucose group and the
coumaroyl-rutinoside chain.
15
Supplemental Table 1: Methyltransferase activity in E.coli cells expressing DIFe1 (MT2).
Crude
Homogenate1
Products3
Substrates
D3R
D3G
none
+
-
none
-
EV
14
C-SAM
Petunidin
Malvidin
+
-
-
+
+
-
-
+
-
+
-
-
EV
-
+
+
-
-
EV
+
-
-
-
-
EV
-
+
-
-
-
EV
-
-
+
-
-
DIFe1
+
-
+
+
+
DIFe1
-
+
+
+
+
DIFe1
+
-
-
-
-
DIFe1
-
+
-
-
-
1
EV, empty vector
2
D3R, delphinidin 3-rutinoside; D3G, delphinidin 3-glucoside: 14C-SAM
3
products detected after acid hydrolysis and TLC
16
Supplemental Table 2. Genotypes of inbred Petunia lines and derived F1 hybrids.
Genotypeb
Linea
AN1
AN2
AN11
HF1
HF2
RT
GF
MT2
MF1 MF2
R27
+/+
+/+
+/+
-/-
-/-
-/-
+/+
+/+
x/x
x/x
W225
-/-
+/+
+/+
-/-
-/-
-/-
+/+
+/+
x/x
x/x
W134
+/+
+/+
-/-
-/-
-/-
-/-
+/+
+/+
x/x
x/x
W240
+/+
m/m
+/+
+/+
../..
+/+ +/+
+/+
x/x
x/x
W240-Rev
+/+
Rev/m +/+
+/+
../..
+/+ +/+
+/+
x/x
x/x
V23
+/+
+/+
+/+
+/+
+/+
+/+ +/+
+/+
+/+
-/-
R51
+/+
+/+
+/+
-/-
-/-
-/-
+/+
+/+
x/x
x/x
V23xR51
+/+
+/+
+/+
+/-
+/-
+/-
+/+
+/+
+/x
+/x
V30
+/+
+/+
+/+
+/+
+/+
+/+ +/+
+/+
-/-
-/-
V32
+/+
+/+
+/+
+/+
+/+
+/+ +/+
-/-
-/-
-/-
M1
+/+
+/+
+/+
-/-
-/-
+/+ +/+
+/+
+/+
+/+
R4
+/+
+/+
+/+
-/-
-/-
-/-
+/+
+/+
+/+
+/+
R43
+/+
+/+
+/+
-/-
-/-
-/-
+/+
-/-
-/-
-/-
R46
+/+
+/+
+/+
-/-
-/-
-/-
+/+
-/-
-/-
-/-
R63
+/+
+/+
+/+
-/-
-/-
+/+ -/-
+/+
-/-
-/-
R78
+/+
+/+
+/+
-/-
-/-
+/+ -/-
-/-
-/-
-/-
R78xV32
+/+
+/+
+/+
+/-
+/+ +/-
-/-
-/-
-/-
R100
+/+
+/+
+/+
-/-
-/-
-/-
+/+
+/+
-/-
+/+
Pi S6
+/+
+/+
+/+
+/+
../..
+/+ +/+
+/+
x/x
x/x
Pa S1
+/+
-/-
+/+
-1/-1 -/-
+/+ +/+
-/-
-/-
-/-
Pa S2
+/+
-/-
+/+
-1/-1 -/-
+/+ +/+
+/+
-/-
-/-
Pa S26
+/+
-/-
+/+
../..
+/+ +/+
-/-
-/-
-/-
a
../..
Lines R27, W225 and W134 are in an isogenic background (R27); W240-Rev is a germinal revertant from
the inbred line W240 that arose by excision of the dTPH1 insertion in an2.
b
Details on W225, W134, W240 and W240-Rev can be found elsewhere (Spelt et al., 2000; Verweij et al.,
2008). Genotypes of all other lines were determined by Wiering, de Vlaming and co-workers in the 1970s.
-/- denotes a homozygous recessive, +/+ homozygous dominant, +/- a heterozygous condition. m/m,
homozygous recessive condition for an unstable an2 allele with a transposon insertion, and m/Rev
heterozygous for a (sporogenic) revertant AN2 allele. z/z,. x/x homozygous dominant condition for MF1
and/or MF2. The prefixes Pi and Pa denote inbred lines of P. inflata and P. axillaris respectively; all other
lines and F1 hybrids are P. hybrida.
17
Supplemental Table 3. Primers used for amplification of MT2, MF1 and MF2 alleles
Gene
Species
Forward primer
Reverse primer
MT2
Ph, Pi, Pa
ATATTACTACCAAGGATTTTACTGAAGC
GCTTGACTTGGCATTGCA
MF1
Ph, Pi
TTCGTCTTAATCTTCTAATTAACCTC
TCACGCTTTCTTTAGCAGATGGA
MF1
Pa
GTTAGGTATCATTACTATTTCCTTCT
GGCCTCCACCTTCCCAGCA
MF2
Ph, Pi
TTTTCGTTTTAATCTTCTAATTAACATTT
CTACTATAAAGATAGTGGATTCCGA
MF2
Pa
GAGGGTCACGGTTTTTGTTCGT
TACTCATTACCCAGTGTTATCCA
Supplemental Table 4. Primer combinations used for quantitative RT-PCR.
Gene
Forward primer
Reverse primer
MT2
ATATTACTACCAAGGATTTTACTGAAGC
GCTTGACTTGGCATTGCA
MF1
CAAGAAGTATATCTTTGAAACTAGTGTT
CTAGGAGAGACGCCTGCAAA
MF2
AGAAGTATATCTTCGAAACTAGTGTG
CTAGCTGAGACGCCTGCC
35S:VvAOMT1
CACTAGTGATATCACAAGTTTGTACA
CTAATAGAGGCGCCTGCAGAG
35S:MT2
CACTAGTGATATCACAAGTTTGTACA
CTAGGAGAGACGCCTGCAAA
35S:MF1
CACTAGTGATATCACAAGTTTGTACA
CTAGCTGAGACGCCTGCC
ACTIN
AGATCTGGCATCATACCTTCTACA
CCMGCAGCTTCCATRCCAATCA
GAPDH
GGTCGTTTGGTTGCAAGAGT
CTGGTTATTCCATTACAACTAC
Supplemental Table 5. Primers used for Real-Time PCR
gene
forward primer
reverse primer
VvAOMT1
GATGAATGTCCCTGTCGATGAG
GCAAGAGCTGTTGCCAAGAGA
MT2
GCAGCGAGCTTTATGGGTCTT
TGAGCTTTAGGAACATCGATAGAAAC
MF1+MF21
CTTTTGGCTACTGCTCTTGCATT
CTCTGTCCGGATCAATCGCTAT
ACTIN
AGCCAACAGAGAGAAGATGACCCA
ACACCATCACCAGAGTCCAACACA
DFRa
TGGAACTGCCAAAGGCTGATA
CTCCTTGACAGCCTTGAATGG
CHSa
CAACTAGTGGTGTGGACATGCC
CACCAGCAAAGCAACCTTGTT
1
These primers do not distinguish between MF1 and MF2 transcripts
Supplemental Table 6. Primers used for construction of fusion genes
primer
sequence1
VvAOMT1-F+attB1
ggggacaagtttgtacaaaaaagcaggct cc ATGTCCAGCTCAAGTCATA
VvAOMT1-R+attB2
ggggaccactttgtacaagaaagctgggt ACTAATAGAGGCGCCTGCAGA
MT2-Fw+attB1
ggggacaagtttgtacaaaaaagcaggct CAATGACAGGCAAAACCGCC
MT2-R+attB2
ggggaccactttgtacaagaaagctgggt ACTAGGAGAGACGCCTGCAAA
MF-F+Topo site
cacc ATGGCAGGCAAAAGCG
MF-R
CTAGCTGAGACGCCTGCC
1
lowercase letters denote attB or Topo sites
18
Supplemental Table 7: Sequence alignment used for the phylogenetic tree in Figure S1 (Fasta format).
>AmBAMT(AAF98284.1)
----MKVMKKLLCMNIAGDGETSYANNSGLQKVMMSKSLHVLDETLKDIIGDHVGFPKCFKMM-DMG------CSSGPNA
LLVM-------SGIINTIEDLYTEKNINELPEFEVFLNDLPDNDFNNL----FKLLSH-----EN---------GNCFVY
GLPG-----SFYGRLLPKKS-----LHFAYSSYSIHWLSQVPEGLEDNNRQNIYMATESPPEVYKAYAKQ-----------YERDFSTFLKLRGEEIVPGGRMVLTFNGRSVEDPSSKDDLAIFTLLAKTLVDMVAEGLVKMDDLYSFNIPIYSPCTRE
VEAAILSEGSFTLDRLEVFRVCWDASDYTDDDDQQDPSIFGKQRSGKFVADCV----RAITEPMLASHFGSTIMDLLFGK
YAKKIVEHLSVENS---------SYFSI-VVSLSRR->At1g19640(AAG23343.1)
------MEVMRVLHMNKGNGETSYAKNSTAQSNIISLGRRVMDEALKKLMMSNSEISSI--GIADLG------CSSGPNS
LLSI-------SNIVDTIHNLCPDLDR-PVPELRVSLNDLPSNDFNYI----CASLPEFYDRVNN--NKEGLGFGRGGGE
SCFVSAVPGSFYGRLFPRRS-----LHFVHSSSSLHWLSQVPCREA-EKEDR--TITADLENMGKIYISKTSPKAHKAYA
LQFQTDFLVFLRSRSEELVPGGRMVLSFLGRRSLDPTTEESCYQWELLAQALMSMAKEGIIEEEKIDAFNAPYYAASSEE
LKMVIEKEGSFSIDRLEISPIDWEGGSISEES--YDLAI------RSKPEALA---SGRRVSNTIRAVVEPMLEPTFGEN
VMDELFERYAKIVGEYFYVSSPRYAIV--ILSLVRTG>CbSAMT(AF133053.1)
------MDVRQVLHMKGGAGENSYAMNSFIQRQVISITKPITEAAITALYSGDTVTTRL--AIADLG------CSSGPNA
LFAV-------TELIKTVEELRKKMGRENSPEYQIFLNDLPGNDFNAI----FRSLPIENDVD-----------GVCFIN
GVPG-----SFYGRLFPRNT-----LHFIHSSYSLMWLSQVPIGIE-SNKGNIYMANTCPQSVLNAYYKQ-----------FQEDHALFLRCRAQEVVPGGRMVLTILGRRSEDRASTECCLIWQLLAMALNQMVSEGLIEEEKMDKFNIPQYTPSPTE
VEAEILKEGSFLIDHIEASEIYWSSCTKDGDGGGSVEEE------GYNVARCM------------RAVAEPLLLDHFGEA
IIEDVFHRYKLLIIERMSKEKTKFINVI--VSLIRKSD
>Os08g0498100(NP001062142.1)
---------------MTTGNGDAPVIKNAHSDIDSTNKTLLKSDALYKYVLDTTVLPREPECMRDLR----LITDKHQWG
FMQS---SADEAQLLGMLLKMAGAKRTIEV-------------GVFTG----YSLLATALALPED---------GKVVAI
DPDR---ESYEIGRPFLEKAGVAHKVD-FREGKGLEKLD--ELLAEEAAAGREAAFDFAFVDADK------------------PNYVKYHEQLLQLVRVGGHIV-----------------------------YDNTLWAGTV----------ALPPDT
PLSDLDRRFSV-----------------------AIRDL------NSRLAADP----------------------------RIDVCQLAIADG----------------ITICRRLV
>Os06g0165800(NP001056910.1)
----MAEAASAAAAATTEQANGSSGGEQKTRHSEVGHKSLLKSDDLYQYILETSVYPREHECMKELR----EVTANHPWN
LMTT---SADEGQFLNLLLKLIGAKKTMEI-------------GVYTG----YSLLATALAIPDD---------GTILAM
DINR---ENYELGLPSIEKAGVAHKID-FREGPALPVLD--QLVEE-EGNHG--SFDFVFVDADK------------------DNYLNYHERLMKLVKVGGLVG-----------------------------YDNTLWNGSV----------VLPADA
PMRKYIRYYRD-----------------------FVLEL------NKALAADH----------------------------RVEICQLPVGDG----------------ITLCRRVK
>ZmCCoAMT(CAB45149.1)
MATTATEATKTTAPAQEQQANGNGNGEQKTRHSEVGHKSLLKSDDLYQYILDTSVYPREPESMKELR----EITAKHPWN
LMTT---SADEGQFLNMLIKLIGAKKTMEI-------------GVYTG----YSLLATALALPED---------GTILAM
DINR---ENYELGLPCINKAGVGHKID-FREGPALPVLD--DLVAD-KEQHG--SFDFAFVDADK------------------DNYLNYHERLLKLVRPGGLIG-----------------------------YDNTLWNGSV----------VLPDDA
PMRKYIRFYRD-----------------------FVLAL------NSALAADD----------------------------RVEICQLPVGDG----------------VTLCRRVK
>ZmCCoAOMT1(ACG24581.1)
------MATTATEAAPAQEQQANGNGEQKTRHSEVGHKSLLKSDDLYQYILDTSVYPREPESMKELR----EVTAKHPWN
LMTT---SADEGQFLNMLIKLIGAKKTMEI-------------GVYTG----YSLLATALALPED---------GTILAM
DINR---ENYELGLPCIEKAGVAHKID-FREGPALPVLD--DLIAE-EKNHG--SFDFVFVDADK------------------DNYLNYHERLLKLVKLGGLIG-----------------------------YDNTLWNGSV----------VLPDDA
PMRKYIRFYRD-----------------------FVLVL------NKALAADD----------------------------RVEICQLPVGDG----------------VTLCRRVK
>ZvCCoAMT(AAA59389.1)
-------------------MATPTGETQPAKHQEVGHKSLLQSDALYQYILETSVYPREPQPMKELR----RITAKHPWN
LMTT---SADEGQFLNLLLKLINAKNTMEI-------------GVYTG----YSLLSTALALPED---------GKILAL
DINR---ENYEIGLPIIQKAGVAHKID-FREGPALPLLD--QMLQD-EKCHG--SFDFIFVDADK------------------DNYLNYHKRLIDLVKFGGVIG-----------------------------YDNTLWNGSL----------VAPADA
PLRKYVRYYRD-----------------------FVLEL------NKALAVDP----------------------------RVEICQLPVGDG----------------ITLCRRIS
>At4g34050(AAL09793.1)
-----MATTTTEATKTSSTNGEDQKQSQNLRHQEVGHKSLLQSDDLYQYILETSVYPREPESMKELR----EVTAKHPWN
IMTT---SADEGQFLNMLIKLVNAKNTMEI-------------GVYTG----YSLLATALALPED---------GKILAM
DVNR---ENYELGLPIIEKAGVAHKID-FREGPALPVLD--EIVAD-EKNHG--TYDFIFVDADK------------------DNYINYHKRLIDLVKIGGVIG-----------------------------YDNTLWNGSV----------VAPPDA
PMRKYVRYYRD-----------------------FVLEL------NKALAADP----------------------------RIEICMLPVGDG----------------ITICRRIS
>MsCCoAOMT(Q40313)
-----------------MATNEDQKQTESGRHQEVGHKSLLQSDALYQYILETSVFPREHEAMKELR----EVTAKHPWN
IMTT---SADEGQFLSMLLKLINAKNTMEI-------------GVYTG----YSLLATALAIPED---------GKILAM
DINK---ENYELGLPVIKKAGVDHKID-FREGPALPVLD--EMIKD-EKNHG--SYDFIFVDADK------------------DNYLNYHKRLIDLVKVGGVIG-----------------------------YDNTLWNGSV----------VAPPDA
PLRKYVRYYRD-----------------------FVLEL------NKALAVDP----------------------------RIEICMLPVGDG----------------ITICRRIK
>PcCCoAMT(AAA33851.1)
-----------------------MASNGESKHSEVGHKSLLQSDALYQYILETSVYPREPEAMKELR----EVTAKHPWN
LMTT---SADEGQFLNMLLKLINAKNTMEI-------------GVYTG----YSLLATALALPDD---------GKILAM
DINR---ENYEIGLPIIEKAGVGHKID-FREGPALPVLD--HMLED-GKYHG--TFDFVFVDADK------------------DNYINYHKRLIDLVKIGGLIG-----------------------------YDNTLWNGSV----------AQPADA
PMRKYVRYYRD-----------------------FVIEL------NKALAADP----------------------------RIEICMLPVGDG----------------VTLCRRIS
>NtCCoAMT5(AAB80931.1)
------------------------MAENGIKHQEVGHKSLLQSDALYQYILETSVYPREPESMKELR----EVTAKHPWN
LMTT---SADEGQFLNMLLKLINAKNTMEI-------------GVYTG----YSLLATALAIPDD---------GKILAM
DINR---ENYEIGLPIIEKAGVAHKIE-FREGPALPVLD--QLVED-KKNHG--TYDFIFVDADK------------------DNYINYHKRIIDLVKVGGLIG-----------------------------YDNTLWNGSV----------VAPPDA
PMRKYVRYYRD-----------------------FVLEL------NKALAVDP----------------------------RIEICMLPVGDG----------------ITLCRRIT
>NtCCoAOMT(AAC49913.1)
-------------------------MATNGRHQEVGHKSLLQSDALYQYILETSVYPREPEPMKELR----EITAKHPWN
LMTT---SADEGQFLSMLIKLINAKNTMEI-------------GVFTG----YSLLATAMALPDD---------GKILAM
DINR---ENYEIGLPVIEKAGLAHKIE-FKEGPALPVLD--QMIED-GKYHG--SYDFIFVDADK------------------DNYLNYHKRLIDLVKIGGLIG-----------------------------YDNTLWNGSV----------VAPPDA
PLRKYVRYYRD-----------------------FVLEL------NKALAADS----------------------------RIEICQLPVGDG----------------ITLCRRIS
19
>VvCCoAMT(CAA90969.1)
----------------------MATNQEAGRHQEVGHKSLLQSDALYQYILETSVYPREPESMKELR----ELTAQHPWN
IMTT---SADEGQFLNMLLKLINAKNTMEI-------------GVYTG----YSLLATALALPDD---------GKILAM
DINK---ENYELGLPVIQKAGVAHKID-FKEGPALPVLD--QMIED-GKYHG--SFDFIFVDADK------------------DNYLNYHKRLIDLVKVGGIIG-----------------------------YDNTLWNGSV----------VAPPDA
PLRKYVRYYRD-----------------------FVLEL------NKALAADP----------------------------RIEICMLPVGDG----------------ITLCRRLS
>PtCCoAOMT1(ACC63876.1)
-----------------MATNGEEQQSQAGRHQEVGHKSLLQSDALYQYILETSVYPREPECMKELR----EVTAKHPWN
IMTT---SADEGQFLNMLLKLVNAKNTMEI-------------GVYTG----YSLLATALAIPED---------GKILAM
DINR---ENYELGLPVIQKAGVAHKID-FKEGPALPVLD--QMIED-GKCHG--SFDFIFVDADK------------------DNYINYHKRLIELVKVGGLIG-----------------------------YDNTLWNGSV----------VAPPDA
PMRKYVRYYRD-----------------------FVLEL------NKALAADP----------------------------RIEICMLPVGDG----------------ITLCRRIQ
>PtCCoAOMT(2CAA12200.1)
-----------------MAANGEEQQTQAGRHQEVGHKSLLQSDALYQYILETSVYPREPECMKELR----ELTAKHPWN
IMTT---SADEGQFLNMLLKLINAKNTMEI-------------GVFTG----YSLLATALAIPED---------GKILAM
DINR---ENYELGLPVIQKAGLEHKIE-FKEGPALPVLD--QMIED-GKYHG--TYDFIFVDADK------------------DNYINYHKRLIELVKVGGLIG-----------------------------YDNTLWNGSV----------VAPADA
PMRKYVRYYRD-----------------------FVLEL------NKALAADP----------------------------RIEICMLPVGDG----------------ITLCRRIK
>AtCCoAMTlike(NP564916.2)
------------------------------MANEIPTKGILKSEALKQYIMETSAYPREHELLKELR--KATVQKYGNLS
EMEV---PVDEGHFLSMLVKIMNAKNTIEI-------------GVFTG----YSLLTTALALPED---------GRITAI
DIDK---EAYEVGLEFIKKAGVDHKIN-FIHSDGLKALD--QLVND-KCE-----FDFAFADADK------------------SSYVNFHERLLKLVKVGGIIA-----------------------------FDNTLWFGF-----------VAEDED
GVPEHMREYRA-----------------------ALIEF------NKKLALDP----------------------------RVEVSQISIGDG----------------ITLCRRLV
>VvAOMT3(CBI27641.3)
-------------------------------MSSLIHKGILKTEALTKYILETSAYPREHEQLRGLR--EATVEKQEFWS
IMNV---PVDEGLFISMLLKLMNVKKTIEL-------------GVFTG----YSLLATPLALPQD---------GKARLS
RKEN---KAYQIGVPFIKKPGVEHKIN-FIQSDAMSVLN--DLIAD-GNEEG--TLDFAFVDADK------------------ENYLNYHELLLKLVRVGGIIA-----------------------------YDNTLWFGSV----------ARSEEE
EMMDFERAGRV-----------------------HLMKL------NKFLASDP----------------------------RVELSHLSIGDG----------------IALCRRYQ
>VvAOMT1(ACO52469.1)
-------------------------------MSSSSHRGILKTEALTKYLLETSAYPREHEQLKGLR--EATVEKHKYWS
LMNV---PVDEGLFISMLLKIMNAKKTIEL-------------GVFTG----YSLLATALALPQD---------GKIIAV
DPDK---EAYQTGVPFIKKAGVEHKIN-FIQSDAMSVLN--DLIAD-GKEEG--TLDFAMVDADK------------------ENYLNYHELLLKLVRVGGIIA-----------------------------YDNTLWFGSV----------ARSEEE
EMMDFERAGRV-----------------------HLMKL------NKFLASDP----------------------------RVELSHLSIGDG----------------VALCRRLY
>VvAOMT2(HQ702997)
-------------------------------MSSSSHRGILKTEALTKYLLETSAYPREHEQLKGLR--EATVEKYKYWS
LMNV---PVDEGLFISMLLKIMNAKKTIEL-------------GVFTG----YSLLATALALPQD---------GKIIAV
DPDK---EAYQTGVPFIKKAGVEHKIN-FIQSDAMTVLN--DLIAD-GKEEG--TLDFAMVDADK------------------ENYLNYHELLLKLVRVGGIIA-----------------------------YDNTLWFGSV----------ARSEEE
EMMDFERAGRV-----------------------HLMKL------NKFLASDP----------------------------RVELSHLSIGDG----------------VALCRRLY
>PhMT2(XXXXX)
------------------------------MTGKTAHPGILRSDALRKYILETSVYPREHELLKELT--KASFENYKAAS
FMGL---PQDEAQFLSMFLKLINAKKTLEI-------------GVFTG----YSLLVTALALPED---------GKVIAI
DPDR---EAYEVGLPYIQKAGVEHKIE-FIQSEAVPVLE--KLLSN-EKEAG--TFDFVFIDADK------------------ENYLKYHEIVLKLVKVGGVIG-----------------------------YDNTLWFGTV----------ALSEDD
PMPEGLRALRG-----------------------HVMKV------NSFLATDP----------------------------RVEVAQLSIGDG----------------LTLCRRLS
>StAMTlike(AGC31679.1)
------------------------------MAEKSGHGTILRSEALKKYILETSVYPREHEQLKELR--QASFDKYKIVS
LMGV---PQDEGQFLSMLLKIMNAKKTMEI-------------GVFTG----YSLLVTALALPED---------GKVIAI
DPDK---DAYEVGLPFIKKAGVEHKIQ-FIQSQALPVLE--KLL---EKEEG--TFDFIFIDADK------------------ENYLKYHEIVLKLVKVGGVLG-----------------------------YDNTLWYGTV----------ALSEND
PMPPGLKALRG-----------------------AVREI------NTFLANDP----------------------------RIEMSQLSIGDG----------------LTLCRRLY
>PhMF1(XXXXX)
------------------------------MAGISGHGSILQSEALKKYIFETSVYPREHEQLKELT--QASFDKYKIVS
LMGV---PPDEAQFLSMLLKIMNAKKTMEI-------------GVFTG----YSLLATALALPED---------GKIIAI
DPDR---EAYEVGLPYIQKAGVEHKIE-FIQSEALPVLQ--KLLSN-GDEEG--TFDFIFIDADK------------------ENYLKYHEIVLKLVKVGGVIG-----------------------------YDNTLWFGTV----------ALSDDD
PIPQGLRELRR-----------------------SVLKI------NSFLATDS----------------------------RIELAHLSIGDG----------------LTLGRRLS
>PhMF2(XXXXX)
------------------------------MAGKSGHGSILQSEALKKYIFETSVYPREHEQLKELT--QASFDKYKIVS
LMGV---PPDEAQFLSMLLKIMNAKKTMEI-------------GVFTG----YSLLATALALPED---------GKIIAI
DPDR---EAYEVGLPYIQKAGVEHKIE-FIQSEALPVLQ--KLLSN-GDEEG--TFDFIFIDADK------------------ENYLKYHEIVLKLVKVGGVIG-----------------------------YDNTLWFGTV----------ALSDDD
PIPQGLRELRR-----------------------SVLKI------NSFLATDS----------------------------RIELAHLSIGDG----------------LTLGRRLS
>CycAOMT2(AB618483)
-------------------------MAEKPTLIIRPNKGVLQSEALYDYIVETSVYPRESKFLKEIR----DVTASHPWS
GMSS---NPDAAQLMGILLKLANAKKTIEV-------------GVFTG----YSLLLTALTIPND---------GKILAI
DPDQ---EAYDLGLPVIKRAGVEHKIN-FVAEAALPVLD--KLLID-PENEG--SFDFAFVDADK------------------ENYLNYHERLLKLVKLGGVIV-----------------------------YDNTLWFGTV----------ALPEDS
VKEGW-RHGRK-----------------------TALEV------NKALTSDP----------------------------RIQLAHVPVGDG----------------NVICIRIQ
>CycAOMT1(AB618482.1)
-------------------------MAEKPTLIIRPNKGVLQSEALYDYIVETSVYPRESKFLKENR----DVTASHPWS
GMSS---NPDAAQLMGILLKLANAKKTIEV-------------GVFTG----YSLLLTALTIPND---------GKILAI
DPDQ---EAYDLGLPVIKRTGVEHKIN-FVAEAALPVLD--KLLID-PENEG--SFDFAFVDADK------------------ENYLNYHERLLKLVKLGGVIV-----------------------------YDNTLWFGTV----------ALPEDS
VKEGW-RHGRK-----------------------TALEV------NKALTSDP----------------------------RIQLAHVPVGDG----------------NVICIRIQ
>At4g26220(NP567739.1)
-------------------------------MAKDEAKGLLKSEELYKYILETSVYPREPEVLRELR----NITHNHPQA
GMAT---APDAGQLMGMLLNLVNARKTIEV-------------GVFTG----YSLLLTALTLPED---------GKVIAI
DMNR---DSYEIGLPVIKKAGVEHKID-FKESEALPALD--ELLNN-KVNEG--GFXXAFVDADK------------------LNYWNYHERLIRLIKVGGIIV-----------------------------YDNTLWGGS-----------VAEPDS
20
STPEWRIEVKK-----------------------ATLEL------NKKLSADQ----------------------------RVQISQAALGDG----------------ITICRRLY
>McPFOMT(AAN61072.1)
--------------------------MDFAVMKQVKNTGLLQSEELCQYILRTSVYPREAGFLKELR----EANESHPDS
YMST---SPLAGQLMSFVLKLVNAKKTIEV-------------GVFTG----YSLLLTALSIPDD---------GKITAI
DFDR---EAYEIGLPFIRKAGVEHKIN-FIESDAMLALD--NLLQG-QESEG--SYDFGFVDADK------------------PNYIKYHERLMKLVKVGGIVA-----------------------------YDNTLWGGT-----------VAQPES
EVPDFMKENRE-----------------------AVIEL------NKLLAADP----------------------------RIEIVHLPLGDG----------------ITFCRRLY
>StlCCoAMT(AAB61680.1)
----------------------MLTKTMGNFFTEVKDTGLLQSEQLHQYILDTSVFPRESEHLKELR----KATESHPMS
FMGT---SPLAGQLLSFMLKTVKPKKTIEV-------------GVFTG----YSLLATALSIPDD---------GKITAV
DIDR---EAYNVGLALIKKAGVESKIS-FIVSDAMTLLD--DLLAD-GRYQG--SYDFAFVDADK------------------TNYVNYHERLIELVKVGGIIA-----------------------------YDNTLWGGT-----------VALPES
EVPDFMKNNWV-----------------------CVTKL------NEILGSDA----------------------------RIDIAHLPVGDG----------------ITFCRRVY
>MsIlOMT(AAB48059.1)
----------MGNSYITKEDNQISATSEQTEDSACLSAMVLTTNLVYPAVLNAAIDLNLFEIIAKATPPGAFMSPSEIAS
KLPASTQHSDLPNRLDRMLRLLASYSVLTSTT-----RTIEDGGAERV----YGLSMVGKYLVPD---ESRGYLASFTTF
LCYPALLQVWMNFKEAVVDED----IDLFKNVHGVTKYE--FMGKD-KKMNQ--IFNKSMVDVCATEMKR-----------MLEIYTGF-EGISTLVDVGGGSGRNLELIISKYPLIKGINFDLPQVIENAPPLSGIEHVGGDMFAS------VPQGDA
MILKAVCHNWSDEKCIEFLSNCHKALSPNG----KVIIV------EFILPEEP--NTSEESKLVSTLDNLMFI-TVGGRE
RTEKQYEKLSKLSGFSKFQVACRAFNSLGVMEFYK-->MclAOMT(M87340)
-------------MTTYTNGNYTQPKTLDKDEQLAGLAVTLANAAAFPMILKSAFELKILDIFSKAG-EGVFVSTSEIAS
QIGA--KNPNAPVLLDRMLRLLASHSVLTC------KLQKGEGGSQRV----YGPAPLCNYLASN---DGQGSLGPLLVL
HHDKVMMESWFHLNDYILEGG----VP-FKRAHGMIQFD--YTGTD-ERFNH--VFNQGMAHHTILVMKK-----------LLDNYNGF-NDVKVLVDVGGNIGVNVSMIVAKHTHIKGINYDLPHVIADAPSYPGVEHVGGNMFES------IPQADA
IFMKWVLHDWSDEHCVKILNKCYESLAKGG----KIILV------ESLIPVIP--EDNLESHMVFSLDCHTLVHNQGGKE
RSKEDFEALASKTGFSTVDVICCAYDTW-VMELYKK->NtCC0AMT(AAL91506.1)
----------------MESSTKSQIPTQSEEERNCTYAMQLLSSSVLPFVLHSTIQLEVFEILAKSN--DTKLSASQIVS
QIPN-CTKPEAPTMLNRMLYVLASYSLFTCSIVE---DEKNNGGQKRV----YGLSQVGKFFVKN---ENGASMGPLLAL
LQNKVFINSWFELKDAVLEGG----VP-FDRVHGVHAFE--YPKSD-PKFND--VFNKAMINHTTVVMKK-----------ILENYKGF-ENLKTLVDVGGGLGVNLKMITSKYPTIKGTNFDLPHVVQHAPSYPGVEHVGGDMFES------VPEGDA
IFMKWILHDWSDSHNLKLLKNCYKALPDNG----KVIVV------EAILPVKP--DIDTAVVGVSQCDLIMMAQNPGGKE
RSEEEFRALATEAGFKGVNLICCVCNFW-VMEFCK-->LpCCoAMT(AAD10253.1)
------------------MGSTAADMAASADEDACMFALQLASSSVLPMTLKNAIELGLLEILVAAG--GKSLTPTEVAA
KLPS-AANPEAPDMVDRILRLLASYNVVTC-----LVEEGKDGRLSRS----YGAAPVCKFLTPN---EDGVSMAALALM
NQDKVLMESWYYLKDAVLDGG----IP-FNKAYGMSAFE--YHGTD-PRFNR--VFNEGMKNHSIIITKK-----------LLELYHGF-EGLGTLVDVGGGVGATVAAIAAHYPTIKGVNFDLPHVISEAPQFPGVTHVGGDMFKE------VPSGDT
ILMKWILHDWSDQHCATLLKNCYDALPAHG----KVVLV------QCILPVNP--EANPSSQGVFHVDMIMLAHNPGGRE
RYEREFQALARGAGFTGVKSTYIY-NAW-AIEFTK-->TaOMT(ABB03907.1)
----------------------MGSIAAGADEDACMYALQLVSSSILPMTLKNAIELGLLETLMAAG--GKFLTPAEVAA
KLPS-AANPEAPDMVDRMLRLLASYNVVSC-----RTEDGKDGRLSRR----YGAAPVCKYLTPN---EDGVSMSALALM
NQDKVLMESWYYLKDAVLDGG----IP-FNKAYGMSAFE--YHGTD-PRFNR--VFNEGMKNHSIIITKK-----------LLESYKGF-EGLGTLVDVGGGVGATVAAITAHYPTIKGINFDLPHVISEAPPFPGVTHVGGDMFQK------VPSGDA
ILMKWILHDWSDEHCATLLKNCYDALPAHG----KVVLV------ECILPVNP--EATPKAQGVFHVDMIMLAHNPGGRE
RYEREFEALAKGAG-AAMKTTYIYANAW-AIEFTK-->CaOMT(AAA80579)
----------------------------------MLFAMQLASASVLPMVLKSAIELDLLEIIASQD---TCMSPTEIAS
HLPT--TNPHAPTMIDRILRLLSSYSIVTC--------SVRSVDDQRV----YSPAPVCKYLTKN---QDGVSIAALCVA
AQDKVLMECWYHMKDAVLDGG----IP-FNKAYGMPIFD--YFAKD-LGSNK--VFNKGMSDFSSMIIKK-----------ILETYKGF-QGLTSLVDVGGGTGATLTKILSKYPTIRGINFDLPHVIQDAPEYPGIEHVGGDMFVS------VPKGDA
IFMKWICHDWNEEQCLKLLKNCYDALPNNG----KVIVA------EYILPVVP--DSSLASKLSVTADVMIVTQNSGGKE
RTEKEFEALAKAAGFQGFQVFCNAFTIY-IIEFSNISN
>TbCatOMT(AAD29844.1)
--------------MGSTENNHNLTPAEEEEEEACLHAMQLASASVLPMVLKAAIELDVLEIIAKAG-KGAYVAPSEIAS
QLST--SNSQAPTVLDRMLRLLASYKVLTC-----NLRNLEDGGVERL----YGLAPVCKFLVKN---EDGVSMAPLVLM
NQDKVLMESWYHLKDAVLDGG----IP-FNKAYGMTAFE--YHGTD-PRFNK--VFNRGMADHSTITMKK-----------LLELYKGF-EGLKSVVDVGGGTGATVNMIVTKHPTIKGINFDLPHVIDDAPAYPGVEHIGGDMFVS------VPKGDA
IFMKWILHDWSDEHSVKFLKNCYESIPADG----KVIIV------ESVLPVFP--ETNLAAHTCFQLDNIMLAHNPGGKE
RTEKDFKALSVKAGFTGFKVVCGAFGSW-VMEFCK-->AtOMT1(AAB96879.1)
---------------MGSTAETQLTPVQVTDDEAALFAMQLASASVLPMALKSALELDLLEIMAKNG---SPMSPTEIAS
KLPT--KNPEAPVMLDRILRLLTSYSVLTC-----SNRKLSGDGVERI----YGLGPVCKYLTKN---EDGVSIAALCLM
NQDKVLMESWYHLKDAILDGG----IP-FNKAYGMSAFE--YHGTD-PRFNK--VFNNGMSNHSTITMKK-----------ILETYKGF-EGLTSLVDVGGGIGATLKMIVSKYPNLKGINFNLPHVIEDAPSHPGIEHVGGDMFVS------VPKGDA
IFMKWICHDWSDEHCVKFLKNCYESLPEDG----KVILA------ECILPETP--DSSLSTKQVVHVDCIMLAHNPGGKE
RTEKEFEALAKASGFKGIKVVCDAFGVN-LIELLKKL>MsCCoAOMT(AAB46623)
---------------MGSTGETQITPTHISDEEANLFAMQLASASVLPMILKSALELDLLEIIAKAG-PGAQISPIEIAS
QLPT--TNPDAPVMLDRMLRLLACYIILTC-----SVRTQQDGKVQRL----YGLATVAKYLVKN---EDGVSISALNLM
NQDKVLMESWYHLKDAVLDGG----IP-FNKAYGMTAFE--YHGTD-PRFNK--VFNKGMSDHSTITMKK-----------ILETYTGF-EGLKSLVDVGGGTGAVINTIVSKYPTIKGINFDLPHVIEDAPSYPGVEHVGGDMFVS------IPKADA
VFMKWICHDWSDEHCLKFLKNCYEALPDNG----KVIVA------ECILPVAP--DSSLATKGVVHIDVIMLAHNPGGKE
RTQKEFEDLAKGAGFQGFKVHCNAFNTY-IMEFLKKV>SlOMT3-AK325603
---------------------MGSTSLTQTEDEAFLFAMQLASASVLPMVLKSAVELELLELMAKAG-PGASISPAELAS
QLPC--KNPDAPVMLDRMLRLLATYSVLNC-----TLRTLPDGRVERL----YSLAPVCKFLTKN---ADGVSVAPLLLM
NQDKVLMQSWYHLKDAVLDGG----IP-FNKAYGMTAFE--YHGTD-PRFNK--VFNRGMSDHTTLSIKK-----------ILEDYKGF-EGLNSIVDVGGGTGATVSMIVSKYPSIKGINFDLPHVIEDAPAYPGVEHIGGDMFVS------VPKADA
IFMKWICHDWSDEHCLKFLKNCYEAVPANG----KVIIA------ECLLPEVP--DTSSSTKNTVHVDVIMLAHNPGGKE
RTEKEFEALAKGAGFNGFTKAC-AYNTW-IMEFTK-->ScOMT(AAO23335.1)
--------------------------MPSSQAQADLWRHTL--YYLTSMGLRCAVKLGIPTAIHNLG---GVSSLPDLAA
ALSI---PASKQPFLGRLMRALVTSGVFAN-------------GKERLLGGLFRLNPLSRILVEGVVAEEHHSQTSFVLA
GTSRHYMEAALGMADWFKKDATGPVPTVFEDVHSASLFDESTAALD-PELDA--LVTEGLAAHDNLGIGT-----------IIREFHDLFKGLVSLTDFCCGDGTTSRAITKAHPHVKFTVLDLPKVIDKTPSDGIVNYFAGDLFHT------VPKAQA
VMLKLVLHHLSYEDCFKILTQCKDAIPSREEGG-KVIVI------DIVVAPSL---GQVMFKEQTLMDILMLV-FTRGRQ
RSENNWHELFTKAGFSDYKIVKKLGARG-VIEVYK-->CrOMT2(AAM97497.1)
---------------------MELQSSEIRNAQAHFFTQVFSFTSMSS--LKCAVQLGIPDAIHSHG---KPMALSDLTN
21
SLPI---NPSKAPYIYRLMRILVAAGYFSE-------------EEKNV----YSLTPFTRLLLKN----DPLNSISMVLG
VNQIAELKAWNAMSEWFQNED----LTAFETAHGKNFWD--FGAED--KYGK--NFDGVMAADSILVSKM-----------LIPEFNYLFEGLDSLVDVGGGTGTIAKAIAKSFPDLKCTVFDLPHVVANLESTENLEFVGGDMFEK------IPSANA
ILLKWILHDWKDEECVKVLKMCRKAIPEKEKGG-KVILI------ETVLMDSKKHENEEAVKAQISSDIDMMV-FFTAKE
RTEEEWATLFREAGFSGYKIFPMIDFRS-PIEVYP-->CrOMT4(AAR02419.1)
---------------------MDLQTAEFREAQAKIWSQAF--SFANCAALKCAVKLGIADAIDNHD--KKALTLSELTE
ELSI---KPSKSPFLQRLMRQLVNAGFFTE--AKQLRDDNKDGRTTTA----YALTPVSRLLLKN----EQWNLRGIVLT
MLDPAELKAWSVLNDWFKNDD----PTAFQTAHEKNYWD--YTAEN-TQHCQ--IFEDAMANDSVLVSKL-----------LVTEYKFLFEGLTSLIDLGGSTGTIAKALAKSFPNLKCTVFDLPHVVANLESTKNLEFVGGDMFEK------LPPSNA
ILLKWILHDWNDEDCVKILKNCKKAIQEKGNGG-KVIII------DTVVYSQK--NEKELVDLQISMDMAMVI-NFAAKE
RTEEEWEHLIREAGFSGHKIFPMYDFRS-IIEVYP-->MpOMT2(AAR09600.1)
----------------MALPNGISSKQELLEAQAHVWNHIY--SYINSMSLKCAIQLGIPDAIHKHG---NPITLSQLAD
ALNI---NKAKSHGLFRLMRILVHSGFFDKVKVKVKVEGEDEEEEEDA----YSLTPASRLLLRS----EPLSVAPFALA
MSDPVYTETWHHLSEWFRNDA----VAAFDTKYGMTFPE--YAVAD-DRLNV--LFNEAMACDAGFVNSI-----------LTTECREIFDGLESMVDVGGGTGATAKGIAAAFPGMECTVLDLPNVVGGLKGSENLSFVSGDMFDF------IPHADA
IFMKFILHDWNDEECVKILKKCKEAISRSNNSCRKIILV------EIVMEDEK--ETHEATETKLFFDMQMLA-IITGKE
RSEKEWGKLFFDAGFTNYKITRVLGLRS-VIEVFP-->VvROMT(CAQ76879.1)
-----------------MDLANGVISAELLHAQAHVWNHIF--NFIKSMSLKCAIQLGIPDIIHNHG---KPMTLPELVA
KLPV---HPKRSQCVYRLMRILVHSGFLAA-------QRVQQGKEEEG----YVLTDASRLLLMD----DSLSIRPLVLA
MLDPILTKPWHYLSAWFQNDD----PTPFHTAHERSFWD--YAGHE-PQLNN--SFNEAMASDARLLTSV-----------LLKEGQGVFAGLNSLVDVGGGTGKVAKAIANAFPHLNCTVLDLPHVVAGLQGSKNLNYFAGDMFEA------IPPADA
ILLKWILHDWSDEECVKILKRCREAIPSKENGG-KVIII------DMIMMKNQ--GDYKSTETQLFFDMTMMIFAP-GRE
RDENEWEKLFLDAGFSHYKITPILGLRS-LIEVYP-->RhOrcMT(AF502433.1)
------MERLNSFRHLNQKWSNGEHSNELLHAQAHIWNHIF--SFINSMSLKSAIQLGIPDIINKHG---YPMTLSELTS
ALPI---HPTKSHSVYRLMRILVHSGFFAK--------KKLSKTDEEG----YTLTDASQLLLKD----HPLSLTPYLTA
MLDPVLTNPWNYLSTWFQNDD----PTPFDTAHGMTFWD--YGNHQ-PSIAH--LFNDAMASDARLVTSV-----------IINDCKGVFEGLESLVDVGGGTGTLAKAIADAFPHIECTVLDLPHVVADLQGSKNLKYTGGDMFEA------VPPADT
VLLKWILHDWSDEECIKILERSRVAITGKEKKG-KVIII------DMMMENQK--GDEESIETQLFFDMLMMA-LVGGKE
RNEKEWAKLFTDAGFSDYKITPISGLRS-LIEVYP-->LjHI4'OMT(BAC58013.1)
---------------MDFSSSNGSEDTELSQAQIHLYKHVY--NFVSSMALKSAMELGIADVIHSHG---KPITLPELAT
ALNL---RPSKIGVLHRFLRLLTHNGFFAK----TTVSRGEGAEEETA----YGLTPPSKLLVKS----NSTCLAPIVKG
ALHPSSLDMWRSSKKWFLEDNEE--LTLFESATGESFWE--FLNKE-TESDTLSMFQEAMAADSH-MFKL-----------ALKECKHVFEGLGSLVDVAGGRGGVTKLIREAFPHVKCTVFDQPQVVANLTGDENLNFVGGDMFKS------VPPADA
VLLKWVLHDWNDELSLKILKNCKEAISGRGKEG-KVIII------DISIDETS--DDRELTELKLDYDLVMLT-MFNGKE
REKKEWEKLIYDAGFSSYKITPICGFKS-LIEVFP-->MsIF7OMT(AAC49928.1)
----------------MASSINGRKPSEIFKAQALLYKHIY--AFIDSMSLKWAVEMNIPNIIQNHG---KPISLSNLVS
ILQV---PSSKIGNVRRLMRYLAHNGFFEI-----------ITKEEES----YALTVASELLVRG----SDLCLAPMVEC
VLDPTLSGSYHELKKWIYEED----LTLFGVTLGSGFWD--FLDKN-PEYNT--SFNDAMASDSKLI-NL-----------ALRDCDFVFDGLESIVDVGGGTGTTAKIICETFPKLKCIVFDRPQVVENLSGSNNLTYVGGDMFTS------IPNADA
VLLKYILHNWTDKDCLRILKKCKEAVTNDGKRG-KVTII------DMVIDKKK--DENQVTQIKLLMDVNMACLN--GKE
RNEEEWKKLFIEAGFQHYKISPLTGFLS-LIEIYP-->GeHI4_OMT(BAC58012.1)
----------------MASSINGRKPSEIFQGQALLYRHIY--AFIDSMCLKWIVELDIPNIIHNHG---KPITVSELVS
ILKV---PQTKAGNVQRIMRYMAHNGFFER------VRIQEEQEENEA----YALTAASELLVKG----SELCLAPMVEC
VLDPTLSGSYHQLKKWIYEED----LTLFGVSLGSHFWE--FLNEN-PEYNK--SFNDAMASDSQMI-NL-----------ALRDCNSGFEGVESIVDVGGGIGTTAKIICDTFPNLKCIVFDRPKVVENLSGTNNLSYVGGDMFQS------VPKADA
VLLKWILHNWTDNDCRRILEKCKEAVSSDGEKG-KVIII------EMVINENQ--DEHEITGTKLLMDVNMACLN--GKE
RSEEEWKKLFIEAGFRDYKISPLTGFLS-LIEVYP---
22
Supplemental Table 8: Sequence alignment used for the phylogenetic tree in Figure 4 (Fasta format).
>Os08g0498100-NP001062142.1
DIDSTNTLLKSDALYKYVLDTTVLPREPECMRDLRLITDKHQWGFMQSSADEAQLLGMLLKMAGAKRTIEVGVFTGYSLLATALALPEDGKVVAIDPDRESYEIGRPFLEKA
GVAHKVDFREGKGLEKLDELLEFDFAFVDADKPNYVKYHEQLLQLVRVGGHIVYDNTLWAGTVALPPDTLSDLDRRFSAIRDLNSRLAADPRIDVCQLAIADGITICRRLV
>Os08g0498600-NP001062144.1
TYRPGSTLLKSDSILEYVLDTTVYPREHERLRELRLITQNHPKSFMGSSPDQMQFFSVLLKMIGARNAVEVGVFTGYSLLATALALPDDGKVVAIDVSREYYELGRPVIEDA
GVAHKVDFRHGDGLAVLDQLLGFDFAYADADKEQYRGYHERLVRLLRVGGVVAYDNTLWGGSVAMPRDTSSAYDRVVRYMVGFNAMVAADDRVEACLLPVADGVTLCRRLK
>Os06g0165800-NP001056910.1
HSEVGHSLLKSDDLYQYILETSVYPREHECMKELREVTANHPWNLMTTSADEGQFLNLLLKLIGAKKTMEIGVYTGYSLLATALAIPDDGTILAMDINRENYELGLPSIEKA
GVAHKIDFREGPALPVLDQLVEFDFVFVDADKDNYLNYHERLMKLVKVGGLVGYDNTLWNGSVVLPADAMRKYIRYYRFVLELNKALAADHRVEICQLPVGDGITLCRRVK
>ZmCCoAMT-CAB45149.1
HSEVGHSLLKSDDLYQYILDTSVYPREPESMKELREVTAKHPWNLMTTSADEGQFLNMLIKLIGAKKTMEIGVYTGYSLLATALALPEDGTILAMDINRENYELGLPCIEKA
GVAHKIDFREGPALPVLDDLIEFDFVFVDADKDNYLNYHERLLKLVKLGGLIGYDNTLWNGSVVLPDDAMRKYIRFYRFVLVLNKALAADDRVEICQLPVGDGVTLCRRVK
>ZmCCoAOMT1-ACG24581.1
HSEVGHSLLKSDDLYQYILDTSVYPREPESMKELREVTAKHPWNLMTTSADEGQFLNMLIKLIGAKKTMEIGVYTGYSLLATALALPEDGTILAMDINRENYELGLPCIEKA
GVAHKIDFREGPALPVLDDLIEFDFVFVDADKDNYLNYHERLLKLVKLGGLIGYDNTLWNGSVVLPDDAMRKYIRFYRFVLVLNKALAADDRVEICQLPVGDGVTLCRRVK
>ZmCCoAMT-CAB45149.1
HSEVGHSLLKSDDLYQYILDTSVYPREPESMKELREITAKHPWNLMTTSADEGQFLNMLIKLIGAKKTMEIGVYTGYSLLATALALPEDGTILAMDINRENYELGLPCINKA
GVGHKIDFREGPALPVLDDLVKFDFAFVDADKDNYLNYHERLLKLVRPGGLIGYDNTLWNGSVVLPDDAMRKYIRFYRFVLALNSALAADDRVEICQLPVGDGVTLCRRVK
>ZmCCoAOMT2-AAP37884.1
HSEVGHSLLKSDDLYQYILDTSVYPREPESMKELREITAKHPWNLMTTSADEGQFLNMLIKLIGAKKTMEIGVYTGYSLLATALALPEDGTILAMDINRENYELGLPCINKA
GVAHKIDFREGPALPVLDDLVKFDFAFVDADKDNYLSYHERLLKLVRPGGLIGYDNTLWNGSVVLPDDAMRKYIRFYRFVLALNSALAADDRVEICQLPVGDGVTLCRRVK
>ZmACF85778.1-ACF85778.1
HSEVGHSLLKSDDLYQYILDTSVYPREPESMKELREITAKHPWNLMTTSADEGQFLNMLIKLIGAKKTMEIGVYTGYSLLATALALPEDGTILAMDINRENYELGLPCINKA
GVGHKIDFREGPALPVLDDLVKFDFAFVDADKDNYLSYHERLLKLVRPGGLIGYDNTLWNGSVVLPDDAMRKYIRFYRFVLALNSALAADDRVEICQLPVGDGVTLCRRVK
>ZvCCoAMT-AAA59389.1
HQEVGHSLLQSDALYQYILETSVYPREPQPMKELRRITAKHPWNLMTTSADEGQFLNLLLKLINAKNTMEIGVYTGYSLLSTALALPEDGKILALDINRENYEIGLPIIQKA
GVAHKIDFREGPALPLLDQMLEFDFIFVDADKDNYLNYHKRLIDLVKFGGVIGYDNTLWNGSLVAPADALRKYVRYYRFVLELNKALAVDPRVEICQLPVGDGITLCRRIS
>At4G34050-NP195131.1
HQEVGHSLLQSDDLYQYILETSVYPREPESMKELREVTAKHPWNIMTTSADEGQFLNMLIKLVNAKNTMEIGVYTGYSLLATALALPEDGKILAMDVNRENYELGLPIIEKA
GVAHKIDFREGPALPVLDEIVEYDFIFVDADKDNYINYHKRLIDLVKIGGVIGYDNTLWNGSVVAPPDAMRKYVRYYRFVLELNKALAADPRIEICMLPVGDGITICRRIS
>PcCCoAMT-AAA33851.1
HQEVGHSLLQSDALYQYILETSVFPREHEAMKELREVTAKHPWNIMTTSADEGQFLSMLLKLINAKNTMEIGVYTGYSLLATALAIPEDGKILAMDINKENYELGLPVIKKA
GVDHKIDFREGPALPVLDEMIEYDFIFVDADKDNYLNYHKRLIDLVKVGGVIGYDNTLWNGSVVAPPDALRKYVRYYRFVLELNKALAVDPRIEICMLPVGDGITICRRIK
>PcCCoAMT-AAA33851.1
HSEVGHSLLQSDALYQYILETSVYPREPEAMKELREVTAKHPWNLMTTSADEGQFLNMLLKLINAKNTMEIGVYTGYSLLATALALPDDGKILAMDINRENYEIGLPIIEKA
GVGHKIDFREGPALPVLDHMLGFDFVFVDADKDNYINYHKRLIDLVKIGGLIGYDNTLWNGSVAQPADAMRKYVRYYRFVIELNKALAADPRIEICMLPVGDGVTLCRRIS
>NtCCoAMT5-AAB80931.1
HQEVGHSLLQSDALYQYILETSVYPREPESMKELREVTAKHPWNLMTTSADEGQFLNMLLKLINAKNTMEIGVYTGYSLLATALAIPDDGKILAMDINRENYEIGLPIIEKA
GVAHKIEFREGPALPVLDQLVKYDFIFVDADKDNYINYHKRIIDLVKVGGLIGYDNTLWNGSVVAPPDAMRKYVRYYRFVLELNKALAVDPRIEICMLPVGDGITLCRRIT
>NtCCoAOMT-AAC49913.1
HQEVGHSLLQSDALYQYILETSVYPREPEPMKELREITAKHPWNLMTTSADEGQFLSMLIKLINAKNTMEIGVFTGYSLLATAMALPDDGKILAMDINRENYEIGLPVIEKA
GLAHKIEFKEGPALPVLDQMIGYDFIFVDADKDNYLNYHKRLIDLVKIGGLIGYDNTLWNGSVVAPPDALRKYVRYYRFVLELNKALAADSRIEICQLPVGDGITLCRRIS
>StCCoAMT-Q8H9B6.1
HQEVGHSLLQSDALYQYILETSVYPREPEAMKELREITAKHPWNLMTTSADEGQFLNMLLKLINAKNTMEIGVFTGYSLLATAMALPDDGKILAMDINRENYEIGLPVIEKA
GLAHKIDFREGPALPVLDQMIGYDFIFVDADKDNYLNYHKRLIDLVKVGGLIGYDNTLWNGSVVAPPDALRKYVRYYRFVLELNKALAADPRIEICQLPVGDGITLCRRIS
>StCCoAOMT-BAC23054.1
HQEVGHSLLQSDALYQYILETSVYPREPEAMKELREITAKHPWNLMTTSADEGQFLNMLLKLINAKNTMEIGVFTGYSLLATAMALPDDGKILAMDINRENYEIGLPVIEKA
GLAHKIDFREGPALPVLDQMIGYDFIFVDADKDNYLNYHKRLIDLVKVGGLIGYDNTLWNGSVVAPPDALRKYVRYYRFVLELNKALAADPRIEICQLPVGDGITLCRRIS
>VvCCoAMT-CAA90969.1
HQEVGHSLLQSDALYQYILETSVYPREPESMKELRELTAQHPWNIMTTSADEGQFLNMLLKLINAKNTMEIGVYTGYSLLATALALPDDGKILAMDINKENYELGLPVIQKA
GVAHKIDFKEGPALPVLDQMIGFDFIFVDADKDNYLNYHKRLIDLVKVGGIIGYDNTLWNGSVVAPPDALRKYVRYYRFVLELNKALAADPRIEICMLPVGDGITLCRRLS
>PtCCoAOMT1-ACC63876.1
HQEVGHSLLQSDALYQYILETSVYPREPECMKELREVTAKHPWNIMTTSADEGQFLNMLLKLVNAKNTMEIGVYTGYSLLATALAIPEDGKILAMDINRENYELGLPVIQKA
GVAHKIDFKEGPALPVLDQMIGFDFIFVDADKDNYINYHKRLIELVKVGGLIGYDNTLWNGSVVAPPDAMRKYVRYYRFVLELNKALAADPRIEICMLPVGDGITLCRRIQ
>PtCCoAOMT-CAA12200.1
HQEVGHSLLQSDALYQYILETSVYPREPECMKELRELTAKHPWNIMTTSADEGQFLNMLLKLINAKNTMEIGVFTGYSLLATALAIPEDGKILAMDINRENYELGLPVIQKA
GLEHKIEFKEGPALPVLDQMIGYDFIFVDADKDNYINYHKRLIELVKVGGLIGYDNTLWNGSVVAPADAMRKYVRYYRFVLELNKALAADPRIEICMLPVGDGITLCRRIK
>At1g67980-NP564916.2
ANEIPTGILKSEALKQYIMETSAYPREHELLKELRKATVQKNLSEMEVPVDEGHFLSMLVKIMNAKNTIEIGVFTGYSLLTTALALPEDGRITAIDIDKEAYEVGLEFIKKA
GVDHKINFIHSDGLKALDQLVKFDFAFADADKSSYVNFHERLLKLVKVGGIIAFDNTLWFGFVAEDEDGVPEHMREYRALIEFNKKLALDPRVEVSQISIGDGITLCRRLV
>At1G67990-NP564917.1
DGRLPDGILKSEALKQYIMETTAYPREHELLKELREATIQRNLSEMGVPVDESLFLSMLVKIINAKNTIEIGVFTGYSLFTVALALPEDGRITAIDIDQAGYNLGLEFMKKA
GVDHKINFIQSDAVRGLDQLLEYDFAFVDADKTNYVYFLEKLLKLVKVGGIIAFDNTLWFGTLIQKENEVPGHMRAYRALLEFNKILARDPRVEIAQISIGDGLTLCRRLI
>PiMT
TGKTAHGILRSDALRKYILETSVYPREHELLKELTKASFEKAASFMGLPQDEAQFLSMFLKLINAKKTLEIGVFTGYSLLVTALALPEDGKVIAIDPDREAYEVGLPYIQKA
GVEHKIEFIQSDAVPVLEKLLEFDFVFIDADKENYLKYHEIVLKLVKVGGVIGYDNTLWFGTVALSEDDMPEGLRALRHVMKVNSFLATDPRVEVAQLSIGDGLTLCRRLS
>PhMT
TGKTAHGILRSDALRKYILETSVYPREHELLKELTKASFENAASFMGLPQDEAQFLSMFLKLINAKKTLEIGVFTGYSLLVTALALPEDGKVIAIDPDREAYEVGLPYIQKA
GVEHKIEFIQSEAVPVLEKLLEFDFVFIDADKENYLKYHEIVLKLVKVGGVIGYDNTLWFGTVALSEDDMPEGLRALRHVMKVNSFLATDPRVEVAQLSIGDGLTLCRRLS
>PhMF1
AGISGHSILQSEALKKYIFETSVYPREHEQLKELTQASFDKIVSLMGVPPDEAQFLSMLLKIMNAKKTMEIGVFTGYSLLATALALPEDGKIIAIDPDREAYEVGLPYIQKA
GVEHKIEFIQSEALPVLQKLLGFDFIFIDADKENYLKYHEIVLKLVKVGGVIGYDNTLWFGTVALSDDDIPQGLRELRSVLKINSFLATDSRIELAHLSIGDGLTLGRRLS
>PhMF2
AGKSGHSILQSEALKKYIFETSVYPREHEQLKELTQASFDKIVSLMGVPPDEAQFLSMLLKIMNAKKTMEIGVFTGYSLLATALALPEDGKIIAIDPDREAYEVGLPYIQKA
GVEHKIEFIQSEALPVLQKLLGFDFIFIDADKENYLKYHEIVLKLVKVGGVIGYDNTLWFGTVALSDDDIPQGLRELRSVLKINSFLATDSRIELAHLSIGDGLTLGRRLS
>PiMF1
AGKGGHSILQSEALKKYIFETSVYPREHEQLKELTQASFDKIVSLMGVPPDEAQFLSMLLKIMNAKKTMEIGVFTGYSLLATALALPEDGKIIAIDPDREAYEVGLPYIQKA
GVEHKIEFVQSEALPVLEKLLGFDFIFIDADKENYLKYHEIVLKLVKVGGVIGYDNTLWFGTVALSDDDIPDGLRELRSVLKINSFLATDPRIELAHLSIGDGLTLGRRLN
>PiMF2
23
AGKSGHSILQSEALKKYIFETSVYPREHEQLKELTQASFDKIVSLMGVPPDEAQFLSMLLKIMNAKKTMEIGVFTGYSLLATALALPEDGKIIAIDPDREAYEVGLPYIQKA
GVEHKIEFIQSEALPVLEKLLGFDFIFIDADKENYLKYHEIVLKLVKVGGVIGYDNTLWFGTVALSDDDIPEGLRELRSVLKINSFLATDPRIELAHLSIGDGLTLGRRLN
>SlCCoAOMTlike-XP004247599.1
AEKSGHSILRSEALKKYILETSVYPREHELLKELTQASFDKIVSLMGVPRDEGQFLSMLLKIMNAKKTMEIGVFTGYSLLTTALALPEDGKVIAIDPDKDAYEVGLPFIKKA
GVEHKIQFIQSQALPVLEKLLEFDFIFIDADKENYLKYHEIVLKLVKVGGVVGYDNTLWFGTVALSEDDMPPGLKALRVVREINTFLANDPRIEMSQLSIGDGLTLCRRLY
>StAMTlike-AGC31679.1
AEKSGHTILRSEALKKYILETSVYPREHEQLKELRQASFDKIVSLMGVPQDEGQFLSMLLKIMNAKKTMEIGVFTGYSLLVTALALPEDGKVIAIDPDKDAYEVGLPFIKKA
GVEHKIQFIQSQALPVLEKLLEFDFIFIDADKENYLKYHEIVLKLVKVGGVLGYDNTLWYGTVALSENDMPPGLKALRAVREINTFLANDPRIEMSQLSIGDGLTLCRRLY
>VvAOMT3-GS
MSSLIHGILKTEALTKYILETSAYPREHEQLRGLREATVEKFWSIMNVPVDEGLFISMLLKLMNVKKTIELGVFTGYSLLATPLALPQDGKARLSRKENKAYQIGVPFIKKP
GVEHKINFIQSDAMSVLNDLIGLDFAFVDADKENYLNYHELLLKLVRVGGIIAYDNTLWFGSVARSEEEMMDFERAGRHLMKLNKFLASDPRVELSHLSIGDGIALCRRYQ
>VvAOMT1-ACO52469.1
MSSSSHGILKTEALTKYLLETSAYPREHEQLKGLREATVEKYWSLMNVPVDEGLFISMLLKIMNAKKTIELGVFTGYSLLATALALPQDGKIIAVDPDKEAYQTGVPFIKKA
GVEHKINFIQSDAMSVLNDLIGLDFAMVDADKENYLNYHELLLKLVRVGGIIAYDNTLWFGSVARSEEEMMDFERAGRHLMKLNKFLASDPRVELSHLSIGDGVALCRRLY
>VvAOMT2-HQ702997
MSSSSHGILKTEALTKYLLETSAYPREHEQLKGLREATVEKYWSLMNVPVDEGLFISMLLKIMNAKKTIELGVFTGYSLLATALALPQDGKIIAVDPDKEAYQTGVPFIKKA
GVEHKINFIQSDAMTVLNDLIGLDFAMVDADKENYLNYHELLLKLVRVGGIIAYDNTLWFGSVARSEEEMMDFERAGRHLMKLNKFLASDPRVELSHLSIGDGVALCRRLY
>Ppppa027001mg-EMJ22806.1
MGDKVEIILKSPALLKYIFETSCFPREHEQLKQLREATVEKFWSLMNVPVDEGLLLSMILKLMNANKTLELGVFTGYSLLTTALAIPHDGKITAIDPDKEAYEFGLPYIQRA
GVDHKINFCHSDALTVLNDLIGFDFAFVDANKDAYIKYHELLLKLVKVGGIIAYDNTLWFGTVVEAEENVEEFAKKGRHLLQLNSFLAADDRIELALVSIGDGLTLCRRLY
>FvCCoAOMTlike-XP004299737.1
HGEEEKIILKSPGLLKYILETSCFPREHELLKQLREATVEKFWSIMNVPVDEGLLLSVLLKIMNAKKTLELGVFTGYSLLTTALALPADGKITAIDPNKEAYEVGLPFIQKA
GVEHKIDFFHSDAFSVLNDLIGFDFVFVDADKDNYINYHELLLKLVKVGGIIAYDNTLWFGTVAEPDEEVEKPLRPGRHVIELNKFLAGDSRVELAFVSIGDGLSLCRRLY
>FvCCoAOMTlike-XP004310071.1
ADSHDHIILKSPALLKYILETSCYPREHELLKQLRDATLEKFWSLMNVPVDEGLLLSMLLKMMNAKKTLELGVFTGYSLLTTALALPNDGKITAIDPDKEAYEFGLPFIQKA
GVDHKINYFASDAFTVLNDLIGFDFAFVDADKENYLNYHELLMKLVKVGGIIAYDNTLWFGTVAQLEENKELALRKGRNVVEVNNFLAADPRVELALVSIGDGLTLCRRLY
>CkmAOMT2-AB618483
AEKPTLGVLQSEALYDYIVETSVYPRESKFLKEIRDVTASHPWSGMSSNPDAAQLMGILLKLANAKKTIEVGVFTGYSLLLTALTIPNDGKILAIDPDQEAYDLGLPVIKRA
GVEHKINFVAEAALPVLDKLLPFDFAFVDADKENYLNYHERLLKLVKLGGVIVYDNTLWFGTVALPEDSVKEGWRHGRTALEVNKALTSDPRIQLAHVPVGDGNVICIRIQ
>CkmAOMT1-AB618482.1
AEKPTLGVLQSEALYDYIVETSVYPRESKFLKENRDVTASHPWSGMSSNPDAAQLMGILLKLANAKKTIEVGVFTGYSLLLTALTIPNDGKILAIDPDQEAYDLGLPVIKRT
GVEHKINFVAEAALPVLDKLLPFDFAFVDADKENYLNYHERLLKLVKLGGVIVYDNTLWFGTVALPEDSVKEGWRHGRTALEVNKALTSDPRIQLAHVPVGDGNVICIRIQ
>At4G26220-NP567739.1
MAKDEAGLLKSEELYKYILETSVYPREPEVLRELRNITHNHPQAGMATAPDAGQLMGMLLNLVNARKTIEVGVFTGYSLLLTALTLPEDGKVIAIDMNRDSYEIGLPVIKKA
GVEHKIDFKESEALPALDELLKFDFAFVDADKLNYWNYHERLIRLIKVGGIIVYDNTLWGGSVAEPDSSTPEWRIEVKATLELNKKLSADQRVQISQAALGDGITICRRLY
>McPFOMT-AAN61072.1
MKQVKNGLLQSEELCQYILRTSVYPREAGFLKELREANESHPDSYMSTSPLAGQLMSFVLKLVNAKKTIEVGVFTGYSLLLTALSIPDDGKITAIDFDREAYEIGLPFIRKA
GVEHKINFIESDAMLALDNLLQYDFGFVDADKPNYIKYHERLMKLVKVGGIVAYDNTLWGGTVAQPESEVPDFMKENRAVIELNKLLAADPRIEIVHLPLGDGITFCRRLY
>StlCCoAMT-AAB61680.1
FTEVKDGLLQSEQLHQYILDTSVFPRESEHLKELRKATESHPMSFMGTSPLAGQLLSFMLKTVKPKKTIEVGVFTGYSLLATALSIPDDGKITAVDIDREAYNVGLALIKKA
GVESKISFIVSDAMTLLDDLLGYDFAFVDADKTNYVNYHERLIELVKVGGIIAYDNTLWGGTVALPESEVPDFMKNNWCVTKLNEILGSDARIDIAHLPVGDGITFCRRVY
24