1
A.
B.
C.
D.
E.
Supplemental Figure S1.Ion fragmentation analysis of anthocyanins present in wild type and b mutant lines. A, Delphinidin-glucose-rhamnose (JI 2822 peak 1, see Fig. 2A). B, Petunidin-glucose-rhamnose (JI 2822 peak 2, see Fig 2A). C, Cyanidin-3-sambubioside-5-glucoside (FN 2271/3/pink, see Fig. 2C). D, Cyanidin-3-rhamnoside-5-glucoside (FN 2271/3/pink, see Fig. 2C). E, Peonidin-3-rhamnoside-5-glucoside peaks (2271/3/pink, see Fig. 2C). Full ion spectra (MS full) and secondary fragmentation spectra (MS2) are shown.
A.
>ADW66160 Pisum sativum
MILILYQTFLLKELSISMFIFFITHFILTFLSKKHHKKLPPGPNGYPFLGALPLMGAMPHLTLFKMSQKYGPIMYLKMGSNNMVVASSPSSAKAFLKTLDQNFSNRPLNAGATHLAYDSQDMVFADYGSRWKLLRKLSNLHMLGGKALEDWSKIREDEMGHMIHTMYDSSKKNESIVLPEMLTYAMANMIGQIILSRRVFETKGCESNEFKDMVVELMTTAGYFNIGDFIPILAWLDLQGIERGMKSLHKKFDALLTKMIEEHVASSHKNPRVKPDFLDTLIAQSKQDSDGGEKLTITNIKAVLLNLFTAGTDTSSSIIEWALAEMLKNPNIMKRAHEEMDRVIGKQRRLQDSDIQNLPYLQAICKETFRKHPSTPLNLPRVSSEACEVNGYYIPKNTRLSVNIWAIGRDPNVWENPLEFNPDRFLSGENAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSFDWKLPNGVVALDMDESFGLALQKKVPLAVVVTPRLSPSAYIP
>CU651565_9 MEDICAGO truncatula
mvitlyqtflykelsislfiflithlffrtfllkinhkklppgpkgypivgalplmgtmphltlfkmsqkygpvmylkmgshnmvvastpssakaflktldlnfsnrppnagathlaydsqdmvfadygsrwkllrklsnlhmlggkaledwskirgdemghmirtmydcskkdesivvsemltyamanmigqvilsrrvfetkgyesnefkdmvvelmttagyfnigdfipilawldlqgiergmkslhkkfdalltkmieehvasslkshrvkpdfldrliaqskedsdgeklsltnikalllnlftagtdtsssiiewalaemlknpkimkkvheemdqvigkqrrfqeadiqklpylqaicketyrkhpstplnlprvslepcqvngyyipkntrlsvniwaigrdpnvwenplefnperfltgknakidprgndfelipfgagrricagtrmgivlvqyilgtlvhsfdwklpngvvdlnmdesfglalqkkvplaaivsprlspsayil
>CU651565_21 MEDICAGO truncatula
mafdafllrdigiaififfishffiglilrntsknlppgprgwpilgvlphlgtmphvtlanmakkfgpimylkmgtcdtvvasssdaaraflktldhnflnrptiigatylgynsqdlvfakygpkwkllrklttlhmlggkalqnwanvrenevkhmvrsihesgkkgesievggllscaitnmvsqvvlskrmfaskgqeskefkemvvefmtisginigdfvpfigwidlegvvgkmkrlhkrfdefltkliedhvncaherkgnpdfldivlandddnfgerlslsnikalllyhfiiltqmylflyiknlftagtdtsssiiewalaemlkspniliraqkemnqvvgrermlvesdleklpylqaicketyrlhpstplsvprvsnkacqingyyipkntrfnvniwaigrdpniwanplefkperflsgkyaridpsgvdfelipfgagrrvcvgykmaivvieyilgtlvhsfdwklpngvelnmdeafgltlekavplsatvtprlvphayv
>LjT34E09_40 LOTUS Japonicus
MASTLDLMFLLREIFVSFFIFLVTRFSFRSLFKSHHQKLPPGPRGWPVLGALPLMGSMPHVTLSNMAKKYGPVMYLKMGTNSLVVASTPAAARAFLKTLDLNFSNRPPNAGATHLAYDSQDMVFAEYGSRWKLLRKLSNLHMLGGKALDDWAQFRDQEMGYMLRAMYDCSKKGEAVVVPEMLTYAMANMIGQVILSRRVFETKGSESNEFKDMVVELMTVAGYFNIGDFIPALGWFDLQGIEGGMKALHKKFDSLLTRMIEEHLASSHKRKLKPDFLDVVMAYQSENSGGEKLSLTNIKALLLNLFTAGTDTSSSIIEWSLAEMLNNPKIMRKAHEEMDRVIGKDRRLRESDLPNLPYFQAICKETYRKHPSTPLNLPRISTEPCEVNGYYIPKNTRLNVNIWAIGRDPDVWENPLEYNPERFLSGRNAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSYDWKLPPGVKELDMEEAFGLALQKKVPLSVMVTPRLPPSAYIP
>BAJ14024 Glycine max CLARK
MDSLLLLKEIATSILIFLITRLSIQTFLKSYRQKLPPGPKGWPVVGALPLMGSMPHVTLAKMAKKYGPIMYLKMGTNNMVVASTPAAARAFLKTLDQNFSNRPSNAGATHLAYDARDMVFAHYGSRWKLLRKLSNLHMLGGKALDDWAQIRDEEMGHMLGAMYDCNKRDEAVVVAEMLTYSMANMIGQVILSRRVFETKGSESNEFKDMVVELMTVAGYFNIGDFIPFLAKLDLQGIERGMKKLHKKFDALLTSMIEEHVASSHKRKGKPDFLDMVMAHHSENSDGEELSLTNIKALLLNLFTAGTDTSSSIIEWSLAEMLKKPSIMKKAHEEMDQVIGRDRRLKESDIPKLPYFQAICKETYRKHPSTPLNLPRISSEPCQVNGYYIPENTRLNVNIWAIGRDPDVWNNPLEFMPERFLSGKNAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVHYILGTLVHSFDWKLPNGERELDMEESFGLALQKKVPLAALVTPRLNPSAYIS
>BAF49293 Clitoria ternatea
MFLLREIGVSILIFMITHLVIRLVLKEKEQRKLPPGPKGWPIVGALPLMGSMPHVTLSEMAKKYGPVMYLKMGTNNMAVASTPSAARAFLKTLDLNFSNRPPNAGATHLAYDAQDMVFADYGSRWKLLRKLSNLHMLGGKALEEWSQVREIEMGHMLRAMYDCSGGGDGNNDNDGNKKKGTRHEPIVVAEMLTYAMANMIGQVILSRRVFETKGSESNEFKDMVVQLMTVAGYFNIGDFIPFLARFDLQGIERGMKTLHNKFDVLLTTMIHEHVASAHKRKGKPDFLDVLMAHHTNESHELSLTNIKALLLNLFTAGTDTSSSIIEWALAEMLINPKIMKKVHEEMDKVIGKDRRLKESDIENLPYLQAICKETYRKHPSTPLNLPRISSQACQVNGYYIPKNTRLSVNIWAIGRDPNVWENPLEFNPERFMGANKTIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSFDWKLPNGVVELNMEETFGLALQKKIPLSALITPRLPPTAYNVINS
>BAE86871 Gentiana scabra
MPIKMSPIYATLTLHLAAALFLFFHVQKLVHYLHGKATGHRLPPGPTGWPILGALPLLGNMPHVTFANMAKKYGPVMYLKVGSHGLAIASTPDAAKAFLKTLDLNFANRPPNAGATHLAYNAQDMVFAHYGPKWKLLRKLSNLHMLGGKALENWADVRKTELGYMLKAMFESSQNNEPVMISEMLTYAMANMLSQVILSRRVFNKKGAKSNEFKDMVVELMTSAGYFNIGDFIPSIGWMDLQGIEGGMKRLHKKFDVLLTRLLDDHERTSHERKQKPDFLDFVIANGDNSDGERLNTDNIKALLLNLFTAGTDTSSSIIEWALAELLKNRTLLTRAQDEMDWVIGRDRRLLESDIPNLPYLQAICKETFRKHPSTPLNLPRIASEPCEVNGYYIPKGTRLNVNIWAIGRDPSVWGDNPNEFDPERFLYGKNAKIDPRGNDFELIPFGAGRRICAGTRMGILLVEYILGTLVHSFDLKLGLPEEELNMDETFGLALQKAVPLAAMLIPRLPLHVYAP
>ABH06585 Vitis vinifera
MAIDTSLLLELAAATLLFFITRFFIRSLLLKSSRKLPPGPKGWPLVGALPLLGNMPHVALAKMAKRYGPVMFLKMGTNSMVVASTPGAARAFLKTLDINFSNRPPNAGATLLAYHAQDMVFADYGARWKLLRKLSNLHMLGGKALEDWSQVRAVELGHMLRAMLELSQRAEPVVVPEMLTFSMANMIGQVILSRRVFETKGSESNEFRDMVVELMTTAGYFNIGDFIPSIAWLDIQGIQRGMKHLHRKFDRLLTKMMEEHTASAHERKGNPDFLDVIMANQENSTGEKLTITNIKALLLNLFTAGTDTSSSVIEWSLAEMLKNPSILKRAHEEMDQVIGRSRRLVESDLPKLPYLQAICKESFRKHPSTPLNLPRVSTQACEVNGYYIPKNTRLSVNIWAIGRDPDVWESPEEFRPERFLSGRNEKIDPRGNDFELIPFGAGRRICAGTRMGIVLVEYILGTLVHSFDWKMPDGVEINMDEAFGLALQKAVSLSAMVTPRLHQSAYAV
>P48418 Petunia HYBrida
MMLLTELGAATSIFLIAHIIISTLISKTTGRHLPPGPRGWPVIGALPLLGAMPHVSLAKMAKKYGAIMYLKVGTCGMAVASTPDAAKAFLKTLDINFSNRPPNAGATHLAYNAQDMVFAHYGPRWKLLRKLSNLHMLGGKALENWANVRANELGHMLKSMSDMSREGQRVVVAEMLTFAMANMIGQVMLSKRVFVDKGVEVNEFKDMVVELMTIAGYFNIGDFIPCLAWMDLQGIEKRMKRLHKKFDALLTKMFDEHKATTYERKGKPDFLDVVMENGDNSEGERLSTTNIKALLLNLFTAGTDTSSSAIEWALAEMMKNPAILKKAQAEMDQVIGRNRRLLESDIPNLPYLRAICKETFRKHPSTPLNLPRISNEPCIVDGYYIPKNTRLSVNIWAIGRDPQVWENPLEFNPERFLSGRNSKIDPRGNDFELIPFGAGRRICAGTRMGIVMVEYILGTLVHSFDWKLPSEVIELNMEEAFGLALQKAVPLEAMVTPRLQLDVYVP
>NP196416 Arabidopsis thaliana tt7
MATLFLTILLATVLFLILRIFSHRRNRSHNNRLPPGPNPWPIIGNLPHMGTKPHRTLSAMVTTYGPILHLRLGFVDVVVAASKSVAEQFLKIHDANFASRPPNSGAKHMAYNYQDLVFAPYGHRWRLLRKISSVHLFSAKALEDFKHVRQEEVGTLTRELVRVGTKPVNLGQLVNMCVVNALGREMIGRRLFGADADHKADEFRSMVTEMMALAGVFNIGDFVPSLDWLDLQGVAGKMKRLHKRFDAFLSSILKEHEMNGQDQKHTDMLSTLISLKGTDLDGDGGSLTDTEIKALLLNMFTAGTDTSASTVDWAIAELIRHPDIMVKAQEELDIVVGRDRPVNESDIAQLPYLQAVIKENFRLHPPTPLSLPHIASESCEINGYHIPKGSTLLTNIWAIARDPDQWSDPLAFKPERFLPGGEKSGVDVKGSDFELIPFGAGRRICAGLSLGLRTIQFLTATLVQGFDWELAGGVTPEKLNMEESYGLTLQRAVPLVVHPKPRLAPNVYGLGSG
>NP001064333 Oryza sativa
MEVAAMEISTSLLLTTVALSVIVCYALVFSRAGKARAPLPLPPGPRGWPVLGNLPQLGGKTHQTLHEMTKVYGPLIRLRFGSSDVVVAGSAPVAAQFLRTHDANFSSRPRNSGGEHMAYNGRDVVFGPYGPRWRAMRKICAVNLFSARALDDLRAFREREAVLMVRSLAEASAAPGSSSPAAVVLGKEVNVCTTNALSRAAVGRRVFAAGAGEGAREFKEIVLEVMEVGGVLNVGDFVPALRWLDPQGVVARMKKLHRRFDDMMNAIIAERRAGSLLKPTDSREEGKDLLGLLLAMVQEQEWLAAGEDDRITDTEIKALILNLFVAGTDTTSTIVEWTMAELIRHPDILKHAQEELDVVVGRDRLLSESDLSHLTFFHAIIKETFRLHPSTPLSLPRMASEECEIAGYRIPKGAELLVNVWGIARDPAIWPDPLEYKPSRFLPGGTHTDVDVKGNDFGLIPFGAGRRICAGLSWGLRMVTMTAATLVHAFDWQLPADQTPDKLNMDEAFTLLLQRAEPLVVHPVPRLLPSAYNIA
>ABH06586 Vitis vinifera
MNPLALFFCTALFCVLLYHFLTRRSVRLPPGPKPWPIVGNLPHLGPVPHHSIAALAKTYGPLMHLRMGFVDVVVAASASVAAQFLKTHDANFSNRPPNSGAKHIAYNYQDLVFAPYGPRWRMLRKICSVHLFSGKALDDFRHIRQEEVAVLTRALARAGQTPVNLGQLLNVCTTNALGRVMLGRRVFGDGSGGEDPKADEFKEMVVELMVLAGVFNIGDFVPALEWLDLQGVAAKMKKLHARFDAFLGAIVEEHKISGSAGSERHVDLLSTLISVRDNADGEGGKLTDVEIKALLLNLFTAGTDTSSSTVEWAIAELIRHPEMMAQAQQELDAVVGRGRLVTDLDLPQLTYLQAIIKETFRLHPSTPLSLPRMAAESCEINGYHIPKNATLLVNVWAIARDPEVWEKPLEFRPSRFLPGGERPNADVRGNDFEVIPFGAGRRICAGMSLGLRMVHLLTATLVHAFNWELPEGQVAEKLNMDEAYGLTLQRAPPLMVHPRPRLSPQVFGK
>BAB83261 Glycine max
MSPLIVALATIAAAILIYRIIKFITRPSLPLPPGPKPWPIVGNLPHMGPVPHHSLAALARIHGPLMHLRLGFVDVVVAASASVAEQFLKIHDSNFSSRPPNAGAKYIAYNYQDLVFAPYGPRWRLLRKLTSVHLFSGKAMNEFRHLRQEEVARLTCNLASSDTKAVNLGQLLNVCTTNALARAMIGRRVFNDGNGGCDPRADEFKAMVMEVMVLAGVFNIGDFIPSLEWLDLQGVQAKMKKLHKRFDAFLTSIIEEHNNSSSKNENHKNFLSILLSLKDVRDDHGNHLTDTEIKALLLNMFTAGTDTSSSTTEWAIAELIKNPQILAKLQQELDTVVGRDRSVKEEDLAHLPYLQAVIKETFRLHPSTPLSVPRAAAESCEIFGYHIPKGATLLVNIWAIARDPKEWNDPLEFRPERFLLGGEKADVDVRGNDFEVIPFGAGRRICAGLSLGLQMVQLLTAALAHSFDWELEDCMNPEKLNMDEAYGLTLQRAVPLSVHPRPRLAPHVYSMSS
>NP182079 cytochrome P450 76C4 Arabidopsis thaliana
MDIISGQALFLLFCFISSCFLISTTARSRRSSGRAATLPPGPPRLPIIGNIHQVGKNPHSSFADLAKIYGPIMSLKFGCLNSVVITSPEAAREVLRTHDQILSGRKSNDSIRCFGHEEVSVIWLPPSSARWRMLRKLSVTLMFSPQRTEATKALRMKKVQELVSFMNESSERKEAVDISRASYTTVLNIISNILFSVDLGSYDSKKSNEFQDTVIGAMEAAGKPDAANYFPFMGFLDLQGNRKAMRGLTERLFRVFRGFMDAKIAEKSLGNYSKDVSNRDFLDSLLILNEGDEAELDNNDIEHLLLDMFTAGTDTSSSTLEWAMAELLRNPKTMVKAQAEMDRVLGQNSVVQESDISGLPYLQAVVKETFRLHPAAPLLVPRKAESDVEVLGFMVPKDTQVLVNVWAIGRDPSVWENPSQFEPERFMGKDIDVKGRDYELTPFGGGRRICPGLPLAVKTVSLMLASLLYSFDWKLPNGVVSEDLDMDETFGITLHRTNTLYAIPVKKQTIN
>NP775426 cytochrome P450 2D10 Rattus norvegicus
MELLNGTGLWPMAIFTVIFILLVDLMHRHQRWTSRYPPGPVPWPVLGNLLQVDPSNMPYSMYKLQHRYGDVFSLQMGWKPMVIVNRLKAVQEVLVTHGEDTADRPPVPIFKCLGVKPRSQGVVFASYGPEWREQRRFSVSTLRTFGMGKKSLEEWVTKEAGHLCDAFTAQNGRSINPKAMLNKALCNVIASLIFARRFEYEDPYLIRMLTLVEESLIEVSGFIPEVLNTFPALLRIPGLADKVFQGQKTFMAFLDNLLAENRTTWDPAQPPRNLTDAFLAEVEKAKGNPESSFNDENLRMVVVDLFTAGMVTTATTLTWALLLMILYPDVQRRVQQEIDEVIGQVRCPEMTDQAHMPYTNAVIHEVQRFGDIAPLNLPRITSCDIEVQDFVIPKGTTLIINLSSVLKDETVWEKPLRFHPEHFLDAQGNFVKHEAFMPFSAGRRACLGEPLARMELFLFFTCLLQHFSFSVPAGQPRPSTLGNFAISVAPLPYQLCAAVREQGH
B.
ABH06586 ------MNPLALFFCTALFCVLLY---HFLTRRSVR--LPPGPKPWPIVGNLPHLG- 45
BAB83261 ------MSPLIVALATIAAAILIYRIIKFITRPSLP--LPPGPKPWPIVGNLPHMG- 48
NP196416 ------MATLFLTILLATVLFLILRIFSHRRNRSHNNRLPPGPNPWPIIGNLPHMG- 50
NP001064333 -MEVAAMEISTSLLLTTVALSVIVCYALVFSRAGKARAPLPLPPGPRGWPVLGNLPQLG- 58
ADW66160 --MILILYQTFLLKELSISMFIFFITHFIL-TFLSKKHHKKLPPGPNGYPFLGALPLMG- 56
CU651565_9 --MVITLYQTFLYKELSISLFIFLITHLFFRTFLLKINHKKLPPGPKGYPIVGALPLMG- 57
LJT34E09_40 --MASTLDLMFLLREIFVSFFIFLVTRFSFRSLFKSHHQ-KLPPGPRGWPVLGALPLMG- 56
BAJ14024 ------MDSLLLLKEIATSILIFLITRLSIQTFLKSYRQ-KLPPGPKGWPVVGALPLMG- 52
BAF49293 ------MFLLREIGVSILIFMITHLVIRLVLKEKEQRKLPPGPKGWPIVGALPLMG- 50
ABH06585 -----MAIDTSLLLELAAATLLFFITRFFIRSLLLKSSR-KLPPGPKGWPLVGALPLLG- 53
BAE86871 MPIKMSPIYATLTLHLAAALFLFFHVQKLVHYLHGKATGHRLPPGPTGWPILGALPLLG- 59
P48418 ------MMLLTELGAATSIFLIAHIIISTLISKTTGRHLPPGPRGWPVIGALPLLG- 50
CU651565_21 -----MAFDAFLLRDIGIAIFIFFISHFFIGLILRNTSK-NLPPGPRGWPILGVLPHLG- 53
NP182079 ----MDIISGQALFLLFCFISSCFLISTTARSRRSSGRAATLPPGPPRLPIIGNIHQVG- 55
NP775426 ----MELLNGTGLWPMAIFTVIFILLVDLMHRHQRWTSR--YPPGPVPWPVLGNLLQVDP 54
ABH06586 PVPHHSIAALAKTYGPLMHLRMGFVDVVVAASASVAAQFLKTHDANFSNRPPNSGAKHIA 105
BAB83261 PVPHHSLAALARIHGPLMHLRLGFVDVVVAASASVAEQFLKIHDSNFSSRPPNAGAKYIA 108
NP196416 TKPHRTLSAMVTTYGPILHLRLGFVDVVVAASKSVAEQFLKIHDANFASRPPNSGAKHMA 110
NP001064333 GKTHQTLHEMTKVYGPLIRLRFGSSDVVVAGSAPVAAQFLRTHDANFSSRPRNSGGEHMA 118
ADW66160 AMPHLTLFKMSQKYGPIMYLKMGSNNMVVASSPSSAKAFLKTLDQNFSNRPLNAGATHLA 116
CU651565_9 TMPHLTLFKMSQKYGPVMYLKMGSHNMVVASTPSSAKAFLKTLDLNFSNRPPNAGATHLA 117
LJT34E09_40 SMPHVTLSNMAKKYGPVMYLKMGTNSLVVASTPAAARAFLKTLDLNFSNRPPNAGATHLA 116
BAJ14024 SMPHVTLAKMAKKYGPIMYLKMGTNNMVVASTPAAARAFLKTLDQNFSNRPSNAGATHLA 112
BAF49293 SMPHVTLSEMAKKYGPVMYLKMGTNNMAVASTPSAARAFLKTLDLNFSNRPPNAGATHLA 110
ABH06585 NMPHVALAKMAKRYGPVMFLKMGTNSMVVASTPGAARAFLKTLDINFSNRPPNAGATLLA 113
BAE86871 NMPHVTFANMAKKYGPVMYLKVGSHGLAIASTPDAAKAFLKTLDLNFANRPPNAGATHLA 119
P48418 AMPHVSLAKMAKKYGAIMYLKVGTCGMAVASTPDAAKAFLKTLDINFSNRPPNAGATHLA 110
CU651565_21 TMPHVTLANMAKKFGPIMYLKMGTCDTVVASSSDAARAFLKTLDHNFLNRPTIIGATYLG 113
NP182079 KNPHSSFADLAKIYGPIMSLKFGCLNSVVITSPEAAREVLRTHDQILSGRKSNDSIRCFG 115
NP775426 SNMPYSMYKLQHRYGDVFSLQMGWKPMVIVNRLKAVQEVLVTHGEDTADRPPVPIFKCLG 114
ABH06586 YN--YQDLVFAPYG-PRWRMLRKICSVHLFS-GKALDDFRHIRQEEVAVLTRALARAGQT 161
BAB83261 YN--YQDLVFAPYG-PRWRLLRKLTSVHLFS-GKAMNEFRHLRQEEVARLTCNLASSDTK 164
NP196416 YN--YQDLVFAPYG-HRWRLLRKISSVHLFS-AKALEDFKHVRQEEVGTLTRELVRVGTK 166
NP001064333 YN--GRDVVFGPYG-PRWRAMRKICAVNLFS-ARALDDLRAFREREAVLMVRSLAEASAA 174
ADW66160 YD--SQDMVFADYG-SRWKLLRKLSNLHMLG-GKALEDWSKIREDEMGHMIHTMYDSS-- 170
CU651565_9 YD--SQDMVFADYG-SRWKLLRKLSNLHMLG-GKALEDWSKIRGDEMGHMIRTMYDCS-- 171
LJT34E09_40 YD--SQDMVFAEYG-SRWKLLRKLSNLHMLG-GKALDDWAQFRDQEMGYMLRAMYDCS-- 170
BAJ14024 YD--ARDMVFAHYG-SRWKLLRKLSNLHMLG-GKALDDWAQIRDEEMGHMLGAMYDCN-- 166
BAF49293 YD--AQDMVFADYG-SRWKLLRKLSNLHMLG-GKALEEWSQVREIEMGHMLRAMYDCSGG 166
ABH06585 YH--AQDMVFADYG-ARWKLLRKLSNLHMLG-GKALEDWSQVRAVELGHMLRAMLELS-- 167
BAE86871 YN--AQDMVFAHYG-PKWKLLRKLSNLHMLG-GKALENWADVRKTELGYMLKAMFESS-- 173
P48418 YN--AQDMVFAHYG-PRWKLLRKLSNLHMLG-GKALENWANVRANELGHMLKSMSDMS-- 164
CU651565_21 YN--SQDLVFAKYG-PKWKLLRKLTTLHMLG-GKALQNWANVRENEVKHMVRSIHESG-- 167
NP182079 HE--EVSVIWLPPSSARWRMLRKLSVTLMFS-PQRTEATKALRMKKVQELVSFMNESS-- 170
NP775426 VKPRSQGVVFASYG-PEWREQRRFSVSTLRTFGMGKKSLEEWVTKEAGHLCDAFTAQN-- 171
ABH06586 P------VNLGQLLNVCTTNALGRVMLGRRVFGDGSGGEDPKADEFKE 203
BAB83261 A------VNLGQLLNVCTTNALARAMIGRRVFNDGNGGCDPRADEFKA 206
NP196416 P------VNLGQLVNMCVVNALGREMIGRRLFG---ADADHKADEFRS 205
NP001064333 PGS------SSPAAVVLGKEVNVCTTNALSRAAVGRRVFA---AGAGEGAREFKE 220
ADW66160 ------KKNESIVLPEMLTYAMANMIGQIILSRRVFE----TKGCESNEFKD 212
CU651565_9 ------KKDESIVVSEMLTYAMANMIGQVILSRRVFE----TKGYESNEFKD 213
LJT34E09_40 ------KKGEAVVVPEMLTYAMANMIGQVILSRRVFE----TKGSESNEFKD 212
BAJ14024 ------KRDEAVVVAEMLTYSMANMIGQVILSRRVFE----TKGSESNEFKD 208
BAF49293 GDGNNDNDGNKKKGTRHEPIVVAEMLTYAMANMIGQVILSRRVFE----TKGSESNEFKD 222
ABH06585 ------QRAEPVVVPEMLTFSMANMIGQVILSRRVFE----TKGSESNEFRD 209
BAE86871 ------QNNEPVMISEMLTYAMANMLSQVILSRRVFN----KKGAKSNEFKD 215
P48418 ------REGQRVVVAEMLTFAMANMIGQVMLSKRVFV----DKGVEVNEFKD 206
CU651565_21 ------KKGESIEVGGLLSCAITNMVSQVVLSKRMFA----SKGQESKEFKE 209
NP182079 ------ERKEAVDISRASYTTVLNIISNILFSVDLGS----YDSKKSNEFQD 212
NP775426 ------GRSINPKAMLNKALCNVIASLIFARRFEY-----EDPYLIRMLT 210
ABH06586 MVVELMVLAGVFNIGDFVPALEWLDLQGVAAKMKKLHARFDAFLGAIVEEHKISG----S 259
BAB83261 MVMEVMVLAGVFNIGDFIPSLEWLDLQGVQAKMKKLHKRFDAFLTSIIEEHNNS-----S 261
NP196416 MVTEMMALAGVFNIGDFVPSLDWLDLQGVAGKMKRLHKRFDAFLSSILKEHEMN------259
NP001064333 IVLEVMEVGGVLNVGDFVPALRWLDPQGVVARMKKLHRRFDDMMNAIIAERRAGSLLKPT 280
ADW66160 MVVELMTTAGYFNIGDFIPILAWLDLQGIERGMKSLHKKFDALLTKMIEEHVAS----SH 268
CU651565_9 MVVELMTTAGYFNIGDFIPILAWLDLQGIERGMKSLHKKFDALLTKMIEEHVAS----SL 269
LJT34E09_40 MVVELMTVAGYFNIGDFIPALGWFDLQGIEGGMKALHKKFDSLLTRMIEEHLAS----SH 268
BAJ14024 MVVELMTVAGYFNIGDFIPFLAKLDLQGIERGMKKLHKKFDALLTSMIEEHVAS----SH 264
BAF49293 MVVQLMTVAGYFNIGDFIPFLARFDLQGIERGMKTLHNKFDVLLTTMIHEHVAS----AH 278
ABH06585 MVVELMTTAGYFNIGDFIPSIAWLDIQGIQRGMKHLHRKFDRLLTKMMEEHTAS----AH 265
BAE86871 MVVELMTSAGYFNIGDFIPSIGWMDLQGIEGGMKRLHKKFDVLLTRLLDDHERT----SH 271
P48418 MVVELMTIAGYFNIGDFIPCLAWMDLQGIEKRMKRLHKKFDALLTKMFDEHKAT----TY 262
CU651565_21 MVVEFMTISG-INIGDFVPFIGWIDLEGVVGKMKRLHKRFDEFLTKLIEDHVNC----AH 264
NP182079 TVIGAMEAAGKPDAANYFPFMGFLDLQGNRKAMRGLTERLFRVFRGFMDAKIAEKSLGNY 272
NP775426 LVEESLIEVSGFIPEVLNTFPALLRIPGLADKVFQGQKTFMAFLDNLLAENRTT-----W 265
ABH06586 AGSERHVDLLSTLISVR----DNADGEG-GKLTDVEIKALLL------NL 298
BAB83261 SKNENHKNFLSILLSLK----DVRDDHG-NHLTDTEIKALLL------NM 300
NP196416 GQDQKHTDMLSTLISLKG---TDLDGDG-GSLTDTEIKALLL------NM 299
NP001064333 DSREEGKDLLGLLLAMVQEQEWLAAGED-DRITDTEIKALIL------NL 323
ADW66160 KNPRVKPDFLDTLIAQS-----KQDSDGGEKLTITNIKAVLL------NL 307
CU651565_9 KSHRVKPDFLDRLIAQS-----KEDSDG-EKLSLTNIKALLL------NL 307
LJT34E09_40 KR-KLKPDFLDVVMAYQ-----SENSGG-EKLSLTNIKALLL------NL 305
BAJ14024 KR-KGKPDFLDMVMAHH-----SENSDG-EELSLTNIKALLL------NL 301
BAF49293 KR-KGKPDFLDVLMAHH-----TNES---HELSLTNIKALLL------NL 313
ABH06585 ER-KGNPDFLDVIMAN------QENSTG-EKLTITNIKALLL------NL 301
BAE86871 ER-KQKPDFLDFVIAN------GDNSDG-ERLNTDNIKALLL------NL 307
P48418 ER-KGKPDFLDVVMEN------GDNSEG-ERLSTTNIKALLL------NL 298
CU651565_21 ER-KGNPDFLDIVLAN------DDDNFG-ERLSLSNIKALLLYHFIILTQMYLFLYIKNL 316
NP182079 SKDVSNRDFLDSLLILN------EGDE-AELDNNDIEHLLL------DM 308
NP775426 DPAQPPRNLTDAFLAEV----EKAKGNPESSFNDENLRMVVV------DL 305
ABH06586 FTAGTDTSSSTVEWAIAELIRHPEMMAQAQQELDAVVGRGRLVTDLDLPQLTYLQAIIKE 358
BAB83261 FTAGTDTSSSTTEWAIAELIKNPQILAKLQQELDTVVGRDRSVKEEDLAHLPYLQAVIKE 360
NP196416 FTAGTDTSASTVDWAIAELIRHPDIMVKAQEELDIVVGRDRPVNESDIAQLPYLQAVIKE 359
NP001064333 FVAGTDTTSTIVEWTMAELIRHPDILKHAQEELDVVVGRDRLLSESDLSHLTFFHAIIKE 383
ADW66160 FTAGTDTSSSIIEWALAEMLKNPNIMKRAHEEMDRVIGKQRRLQDSDIQNLPYLQAICKE 367
CU651565_9 FTAGTDTSSSIIEWALAEMLKNPKIMKKVHEEMDQVIGKQRRFQEADIQKLPYLQAICKE 367
LJT34E09_40 FTAGTDTSSSIIEWSLAEMLNNPKIMRKAHEEMDRVIGKDRRLRESDLPNLPYFQAICKE 365
BAJ14024 FTAGTDTSSSIIEWSLAEMLKKPSIMKKAHEEMDQVIGRDRRLKESDIPKLPYFQAICKE 361
BAF49293 FTAGTDTSSSIIEWALAEMLINPKIMKKVHEEMDKVIGKDRRLKESDIENLPYLQAICKE 373
ABH06585 FTAGTDTSSSVIEWSLAEMLKNPSILKRAHEEMDQVIGRSRRLVESDLPKLPYLQAICKE 361
BAE86871 FTAGTDTSSSIIEWALAELLKNRTLLTRAQDEMDWVIGRDRRLLESDIPNLPYLQAICKE 367
P48418 FTAGTDTSSSAIEWALAEMMKNPAILKKAQAEMDQVIGRNRRLLESDIPNLPYLRAICKE 358
CU651565_21 FTAGTDTSSSIIEWALAEMLKSPNILIRAQKEMNQVVGRERMLVESDLEKLPYLQAICKE 376
NP182079 FTAGTDTSSSTLEWAMAELLRNPKTMVKAQAEMDRVLGQNSVVQESDISGLPYLQAVVKE 368
NP775426 FTAGMVTTATTLTWALLLMILYPDVQRRVQQEIDEVIGQVRCPEMTDQAHMPYTNAVIHE 365
ABH06586 TFRLHPSTPLSLPRMAAESCEINGYHIPKNATLLVNVWAIARDPEVW-EKPLEFRPSRFL 417
BAB83261 TFRLHPSTPLSVPRAAAESCEIFGYHIPKGATLLVNIWAIARDPKEW-NDPLEFRPERFL 419
NP196416 NFRLHPPTPLSLPHIASESCEINGYHIPKGSTLLTNIWAIARDPDQW-SDPLAFKPERFL 418
NP001064333 TFRLHPSTPLSLPRMASEECEIAGYRIPKGAELLVNVWGIARDPAIW-PDPLEYKPSRFL 442
ADW66160 TFRKHPSTPLNLPRVSSEACEVNGYYIPKNTRLSVNIWAIGRDPNVW-ENPLEFNPDRFL 426
CU651565_9 TYRKHPSTPLNLPRVSLEPCQVNGYYIPKNTRLSVNIWAIGRDPNVW-ENPLEFNPERFL 426
LJT34E09_40 TYRKHPSTPLNLPRISTEPCEVNGYYIPKNTRLNVNIWAIGRDPDVW-ENPLEYNPERFL 424
BAJ14024 TYRKHPSTPLNLPRISSEPCQVNGYYIPENTRLNVNIWAIGRDPDVW-NNPLEFMPERFL 420
BAF49293 TYRKHPSTPLNLPRISSQACQVNGYYIPKNTRLSVNIWAIGRDPNVW-ENPLEFNPERFM 432
ABH06585 SFRKHPSTPLNLPRVSTQACEVNGYYIPKNTRLSVNIWAIGRDPDVW-ESPEEFRPERFL 420
BAE86871 TFRKHPSTPLNLPRIASEPCEVNGYYIPKGTRLNVNIWAIGRDPSVWGDNPNEFDPERFL 427
P48418 TFRKHPSTPLNLPRISNEPCIVDGYYIPKNTRLSVNIWAIGRDPQVW-ENPLEFNPERFL 417
CU651565_21 TYRLHPSTPLSVPRVSNKACQINGYYIPKNTRFNVNIWAIGRDPNIW-ANPLEFKPERFL 435
NP182079 TFRLHPAAPLLVPRKAESDVEVLGFMVPKDTQVLVNVWAIGRDPSVW-ENPSQFEPERFM 427
NP775426 VQRFGDIAPLNLPRITSCDIEVQDFVIPKGTTLIINLSSVLKDETVW-EKPLRFHPEHFL 424
ABH06586 PGGERPNADVRGNDFEVIPFGAGRRICAGMSLGLRMVHLLTATLVHAFNWELPEGQVAEK 477
BAB83261 LGGEKADVDVRGNDFEVIPFGAGRRICAGLSLGLQMVQLLTAALAHSFDWELEDCMNPEK 479
NP196416 PGGEKSGVDVKGSDFELIPFGAGRRICAGLSLGLRTIQFLTATLVQGFDWELAGGVTPEK 478
NP001064333 PGGTHTDVDVKGNDFGLIPFGAGRRICAGLSWGLRMVTMTAATLVHAFDWQLPADQTPDK 502
ADW66160 SG-ENAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSFDWKLPNGV--VA 483
CU651565_9 TG-KNAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSFDWKLPNGV--VD 483
LJT34E09_40 SG-RNAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSYDWKLPPGV--KE 481
BAJ14024 SG-KNAKIDPRGNDFELIPFGAGRRICAGTRMGIVLVHYILGTLVHSFDWKLPNGE--RE 477
BAF49293 G--ANKTIDPRGNDFELIPFGAGRRICAGTRMGIVLVQYILGTLVHSFDWKLPNGV--VE 488
ABH06585 SG-RNEKIDPRGNDFELIPFGAGRRICAGTRMGIVLVEYILGTLVHSFDWKMPDG---VE 476
BAE86871 YG-KNAKIDPRGNDFELIPFGAGRRICAGTRMGILLVEYILGTLVHSFDLKLGLPE--EE 484
P48418 SG-RNSKIDPRGNDFELIPFGAGRRICAGTRMGIVMVEYILGTLVHSFDWKLPSEV--IE 474
CU651565_21 SG-KYARIDPSGVDFELIPFGAGRRVCVGYKMAIVVIEYILGTLVHSFDWKLPNG---VE 491
NP182079 GK----DIDVKGRDYELTPFGGGRRICPGLPLAVKTVSLMLASLLYSFDWKLPNGVVSED 483
NP775426 DA-----QGNFVKHEAFMPFSAGRRACLGEPLARMELFLFFTCLLQHFSFSVPAGQ---- 475
ABH06586 LNMDEAYGLTLQRAPPLMVHPRPRLSPQVFGK--- 509
BAB83261 LNMDEAYGLTLQRAVPLSVHPRPRLAPHVYSMSS- 513
NP196416 LNMEESYGLTLQRAVPLVVHPKPRLAPNVYGLGSG 513
NP001064333 LNMDEAFTLLLQRAEPLVVHPVPRLLPSAYNIA-- 535
ADW66160 LDMDESFGLALQKKVPLAVVVTPRLSPSAYIP--- 515
CU651565_9 LNMDESFGLALQKKVPLAAIVSPRLSPSAYIL--- 515
LJT34E09_40 LDMEEAFGLALQKKVPLSVMVTPRLPPSAYIP--- 513
BAJ14024 LDMEESFGLALQKKVPLAALVTPRLNPSAYIS--- 509
BAF49293 LNMEETFGLALQKKIPLSALITPRLPPTAYNVINS 523
ABH06585 INMDEAFGLALQKAVSLSAMVTPRLHQSAYAV--- 508
BAE86871 LNMDETFGLALQKAVPLAAMLIPRLPLHVYAP--- 516
P48418 LNMEEAFGLALQKAVPLEAMVTPRLQLDVYVP--- 506
CU651565_21 LNMDEAFGLTLEKAVPLSATVTPRLVPHAYV---- 522
NP182079 LDMDETFGITLHRTNTLYAIPVKKQTIN------511
NP775426 -PRPSTLGNFAISVAPLPYQLCAAVREQGH----- 504
Supplemental Figure S2.F3’5’H sequence analysis.
A, FASTA files of cytochrome P450 amino acid sequences from eudicot plants with NP775426 from Rattusnorvegicus included as an outgroup comparison. B, ClustalW 2.1 multiple sequence alignment. Absolutely conserved residues are highlighted in bold, cytochrome P450 motifs are underlined. The G305D polymorphism in soybean sequences ABQ96218 and BAJ14024 is highlighted.
1
|....|....|....|....|....|....|....|....|....|....|....|....|....|....
280 290 300 310 320 330 340
JI 2822 GCATTTCTCAAAACACTCGATCAAAATTTCTCAAACCGTCCGCTCAATGCGGGAGCGACACATCTAGCTT
A F L K T L D Q N F S N R P L N A G A T H L A Y
JI 73 GCATTTCTCAAA------CCGTCCGCTCAATGCGGGAGCGACACATCTAGCTT
A F L K P S A Q C G S D T S S L
JI 118 GCATTTCTCAAAACACTCGATCAAAATTTCTCAAACCGTCCGCTCAATGCGGAAGCGACACATCTAGCTT
A F L K T L D Q N F S N R P L N A E A T H L A Y
. | . . . . | . . . . | . . . . | . . . . | . .
95 100 105 110 115
|....|....|....|....|....|....|....|....|....|....|....|....|....|....
350 360 370 380 390 400 410
JI 2822 ACGATTCACAAGACATGGTTTTCGCGGATTACGGATCCAGGTGGAAATTACTTAGAAAACTAAGTAACTTA
D S Q D M V F A D Y G S R W K L L R K L S N L
JI 73 ACGATTCACAAGACATGGTTTTCGCGGATTACGGATCCAGGTGGAAATTACTTAGAAAACTAAGTAACTTA
R F T R H G F R G L R I Q V E I T *
JI 118 ACGATTCACAAGACATGGTTTTCGCGGATTACGGATCCAGGTGGAAATTACTTAGAAAACTAAGTAACTTA
D S Q D M V F A D Y G S R W K L L R K L S N L
. . | . . . . | . . . . | . . . . | . . . . |
120 125 130 135 140
Supplemental Figure S3.Sequence characterization of mutant b alleles.
Nucleotide and amino acid sequences of a wild type allele from line JI 2822, a spontaneous deletion allele from line JI 73 with the 23-bp deletion indicated by dashes and the b mutant type allele from line JI 118 with the single nucleotide polymorphism underlined. Predicted changes in the amino acid sequences of the mutants are italicized.Numbering is according to JI 2822, from the A of the ATG on genomic sequence and the start methionine on the amino acid sequence.
Supplemental Figure S4. Characterisation of mutant b alleles by PCR.
A, PCR products amplified from wild type line JI 2822 and b mutant line FN 1076/6. The B gene is shown as a horizontal line with exons as thick lines and with the ATG start codon labelled. The position of an Ogreretroelement insertion in line FN 1076/6 is indicated by a triangle. Positions of primers are indicated by black arrows with the name of the primer above (see Supplemental Table 1 for primer sequences). PCR amplification from genomic DNA is represented by a connecting line between primers and F3’5’H PCR products, indicated by red arrows, are shown in gel photographs above or below the connecting line. The gel photographs show a 100-bp marker lane in the left lane, PCR products from line JI 2822 in the middle lane and PCR products from line FN 1076/6 in the right lane. PCR products not indicated by a red arrow are controls amplified from Ago primers. B, Diagram of the b gene from line FN 1076/6. The b gene is shown as a horizontal line with exons as thick lines and with the ATG start codon labelled. The position of an Ogreretroelement insertion is indicated by open rectangles. Positions of primers are indicated by black arrows as in panel A. Insertion breakpoints are indicated with vertical curved lines and labeled with nucleotide positions in the b gene and an Ogre element. C, PCR products amplified from wild type line JI 2822 and b mutant lines. The B gene is shown as a horizontal line with exons as thick lines and with the ATG start codon labelled. The position of an Ogreretroelement insertion in line FN 1076/6 is indicated by a triangle. Positions of primers are indicated by black arrows and gel photographs of PCR products are as in panel A. PCR amplification between primers is represented by a colored horizontal line. A red line indicates that no F3’5’H PCR product was amplified from any of the FN lines shown. A green line indicates that a F3’5’H PCR product was amplified from line FN 1076/6, but none of the other FN lines.
Medtr3g169650
44305329
ATGGCTTTTG ATGCGTTTCT TCTAAGAGAT ATTGGTATTG CCATTTTCAT CTTCTTCATC TCACACTTCT TCATAGGCTT
GATTCTTAGA AACACTTCTA AAAATCTTCC ACCTGGCCCA AGAGGATGGC CAATTCTTGG TGTTCTTCCA CACTTGGGAA
CCATGCCTCA TGTCACACTC GCAAACATGG CGAAGAAATT TGGACCGATC ATGTATCTAA AAATGGGCAC TTGTGACACT
GTGGTTGCTT CAAGCTCAGA TGCAGCTAGA GCATTTCTCA AGACTTTAGA CCATAATTTC CTAAATAGGC CTACCATTAT
TGGTGCAACT TATTTAGGTT ACAATTCACA AGACCTTGTT TTTGCAAAAT ATGGACCTAA ATGGAAACTA CTTAGGAAAT
TAACAACCTT ACACATGCTT GGTGGGAAAG CACTTCAAAA TTGGGCTAAT GTGAGAGAAA ATGAGGTGAA ACACATGGTG
AGATCCATTC ATGAGTCTGG GAAAAAAGGC GAATCGATTG AGGTTGGTGG TTTGTTGAGT TGTGCAATAA CAAACATGGT
GAGCCAAGTT GTACTAAGCA AACGTATGTT TGCAAGTAAA GGGCAAGAGT CAAAGGAGTT TAAGGAAATG GTTGTGGAGT
TCATGACAAT ATCTGGAATC AACATTGGTG ATTTTGTACC TTTCATTGGT TGGATTGATT TGGAAGGTGT AGTGGGTAAG
ATGAAACGAT TGCATAAGAG GTTTGATGAG TTTTTGACCA AGTTGATTGA AGATCATGTG AATTGTGCTC ATGAACGTAA
AGGGAATCCA GATTTTTTGG ATATTGTTTT GGCAAATGAT GATGATAATT TTGGTGAGAG ATTGAGCTTG TCCAACATCA
AGGCTTTATT ATTGGTACTT ACTTCTCTCT TTCCCAACTA CTACTTTCTT TATTTTTTAT TTTTTGTGAC CGGAGTTTGA
ACCTCAGACT TTGCATATAT TATGCCTGTC TGTATCAACT AAGTTAAGCT CAGGAGAATA A
44306349
Intron size = 2976 nt (44306223 – 44309199)
Medtr3g169660
44309176
ATGTATCTTT TCCTATATAT CAAGAATTTG TTCACTGCAG GGACAGATAC ATCATCTAGC ATCATAGAGT GGGCACTTGC
TGAAATGCTA AAAAGTCCAA ACATCTTGAT AAGGGCACAA AAAGAGATGA ATCAAGTGGT AGGCAGGGAA AGAATGCTAG
TTGAATCTGA CCTAGAAAAA TTACCATACC TTCAAGCAAT ATGCAAGGAG ACATATCGTT TGCATCCATC TACACCCCTT
AGTGTTCCAA GGGTGTCAAA TAAAGCATGC CAAATTAATG GTTACTACAT CCCCAAAAAT ACAAGGTTCA ATGTTAACAT
ATGGGCAATA GGGAGAGACC CAAATATATG GGCTAACCCA TTAGAATTCA AACCTGAAAG ATTTTTAAGT GGAAAATATG
CAAGAATTGA TCCAAGTGGA GTTGATTTTG AATTGATTCC TTTTGGAGCT GGAAGAAGAG TTTGTGTTGG GTATAAAATG
GCTATTGTGG TGATTGAGTA CATTCTGGGC ACATTGGTAC ACTCTTTTGA CTGGAAATTA CCAAATGGTG TTGAGCTAAA
CATGGATGAG GCCTTTGGAC TTACATTGGA AAAAGCAGTG CCCTTGTCAG CTACGGTTAC CCCTCGCTTG GTCCCACATG
CTTATGTTTA A
44309826
Supplemental Figure S5.Proposed splicing model for medicago gene CU651565_21.
Proposed splicing model for the F3’5’H gene CU651565_21 joins ORFs Medtr3g169650 and Medtr3g169660, as annotated in version 3.0 of medicago genome pseudomolecules. Proposed splice donor and splice acceptor dinucleotides defining a 2976 nt intron are highlighted.
1
Primer Name / Sequence 5’ to 3’pSf35 h5' / CAAACCGTCCGCTCAATGCGGG
pink mt F1 / GGTGATCACCCTATACCAAACGTTCCT
F2 / TAGCTTACGATTCACAAGACATG
F2 comp / CATGTCTTGTGAATCGTAAGCTA
R3 / AGATCCAACCAAGCAAGAATTG
R3comp / CAATTCTTGCTTGGTTGGATCT
5' rose S1 / CGAGTGTTTTGAGAAATGCTTTGGC
5' rose S2 / GAAGGAGATGAGGCAACAACCATGTTG
5' rose S3 / GATAACCATTTGGGCCTGGTG
3'pink s1 / ATTGAGGAGCATGTTGCTTCTAG
3'pink s2 / GAATCTATTTACAGCAGGAACCGATAC
3'pink s3 / CWTCAAGTATMATAGARTGGKCC
pink int-F / GCAAACTTTATAGATCAGAGTCCC
3'extR / CACAATAACAAGGGCTGAAC
Pink-gly-cli-R / CAACCTAGGGGTAATCAAAGCA
5'pinkS1 / GATTGAGGAGCATGTTGCTTCTAG
5'pinkS2 / GAATCTATTTACAGCAGGAACCGATAC
3'roseS2 / GAAGGAGATGAGGCAACAACCATG
5' rose S1 / CGAGTGTTTTGAGAAATGCTTTGGC
3'pink s3 / CWTCAAGTATMATAGARTGGKCC
Pink mt F1 / GGTGATCACCCTATACCAAACGTTCC
3' pink S2 comp / GTATCGGTTCCTGCTGTAAATAGATTC
Pink gly-cli R / CAACCTAGGGGTAATCAAAGCAG
pink intF / GCAAACTTTATAGATCAGAGTCCC
5'roseS3 / GATAACCATTTGGGCCTGGTG
PINK-int-R / GGGACTCTGATCTATAAAGTTTGC
3'pinkS4 / CCTCGAGTTAAGCCTG
3'pinkR5 / CTTGTAACCGTCGCTG
3'pinkS5 / TTCTGATGGAGGAGAG
3'pinkR6 / TGGCGTTGATGGGTG
3'pinkR4 / CTGCTAGGGCCCATTC
pink int F2 / GCATCAGGTACGGTATCG
pink extra5'-F / CCGAGTGTTTTGAGGAACG
5' rose S4 / ATGAAGAAAATGAACATGGAGATGG
5' rose S2 Rc / CAACATGGTTGTTGCCTCATCTCCTTC
5'rose iPCR1 / AGCCATGCCTCATCTCACACTC
MtESTpink-F / GAATCACCACTAATTCAGTTACAAG
S-PCRpinkSspI-F / CAATCTCTTCAACCTCCAATTC
3pinkF / TTACCAAGAGTATCATCAGAAGC
pink pro F / CAATGCACGTCCGTAGAGG
F35H-3endF2 / CCATTTGGTGCTGGGAG
pSf35-F2 / TAGCTTACGATTCACAAGACATG
pSf35-F2 comp / CATGTCTTGTGAATCGTAAGCTA
pSf35-R3 / AGATCCAACCAAGCAAGAATTG
pSf35-R3comp / CAATTCTTGCTTGGTTGGATCT
mtF35HF1 / TATCCAATTGTAGGTGCACTCCC
mtF35HR2 / ACTTGATGATGTATCGGTTCCTGC
PsAGO1 / TTCTACCTCTGTGCTCATGC
PsAGO2 / CCTCGAATTTCATGAATTGC
oligo T primer / GACTCGAGTCGACATCG(T)17
APX1A / CGTCTGATGCTGACTACGCT
APX1B / ACTACGCTTGCATCGTCGCT