1
Additional data
Additional file 1: Figure S1. Meta-expression analysis and genome-wide identification of RopGEF in rice. The expressions from various rice tissues were examined using Microarray
database. Yellow color in the heatmap indicates high level of expression; dark blue, low level of expression. Numeric values indicate the average of the normalized log2 intensity of microarray data.
5 9 13
L e a f b la d e L e a f s h e a th F la g l e a f Ste m An th e r
Roo t In fl o re s c e n c e O v a ry Pi s ti l Pa le a L e m m a Em b ry o En d o s p e rm
Roo t h a ir
OsRopGEF6 OsRopGEF2 OsRopGEF8 OsRopGEF3 OsRopGEF4 OsRopGEF7 OsRopGEF10 OsRopGEF5 OsRopGEF9 OsRopGEF1 OsRopGEF11
Supplementary Fig. S1
2
Additional file 2: Figure S2. Meta-expression analysis of entire AtRopGEF genes. The heatmap was prepared using the Genevestigator. We chose five representative tissues, including pollen. It revealed that five Arabidopsis RopGEFs were highly expressed in pollen. The dark red color of the heatmap indicated the highest expression; white color, lowest expression.
Pollen
Pistill
Seed
Shoot
Root
0% 100%
Supplementary Fig. S2
3
LOC_Os09g37270_OsRopGEF7_ GELWRLEPMASARKAMWTREMDWLLSVADSIVELTPSIQELPDGGGQFEVMVPRPRSDLYMNLPALKKLDAMLLAMIDGFKE-TEFWYVDRG--IVVDDSGGPFSSSSSSCGRPSVRQEE 244 AT4G38430_AtRopGEF1_ GELWRLEPLAPQKKAMWRRELEWLLCVSDSIVELIPSIQQFP-GGGTYEIMETRPRSDLYANLPALKKLDAMLIDMLDAFSD-TEFWYTDRG--IVLGDCD---KDSYNSPASVRQED 243 LOC_Os10g40270_OsRopGEF1_ GSCHKLEPLPAGKKTMWRREMDCLLSVCDYIVEFYPSSQTLP-DGTKVEVMATRPRSDIYINLPALEKLDAMLIEILESFQK-AEFWYADAGTRSFGSATSSSTMSSSSFRRSTH-RNED 270 LOC_Os04g47170_OsRopGEF9_ GGCHRLEPLLAEKRSMWRREMDCLLSVCDYIVELFPSKEIMP-DGTVREVMATRPRSDIYVNLPALEKLDDMLLEILDSFQK-TEFWYVND----KGQKDSCAAAAAAPCRPVSH-RDGD 249 LOC_Os07g29780_OsRopGEF11_ GTCHRLQPLPPEKKAMWNREMDCLLSICEYIVEFSPKVQAMP-DGSTHDVMATSPRSDILMNLPALEKLETMLLEILDSFEK-TDFWYIDQ---RKQSFSDSMKFQ---RSED 233 AT1G01700_AtRopGEF2_ GQNLRLEPLEIEQKTTWKREMNCLLSVCDYIFEFIPKSQNLS-NGATVEVMESRPRADIYINLPALRKLDSMLMEALDSFQK-TEFWYAEEG--SLSMKSTRS--ATGSFRKVIVQRKEE 272 AT4G00460_AtRopGEF3_ GQNLRLEPLETEKRALWKREMNCLLSVCDYIVEFIPRCQNLS-NGATVEVMESRPRADIYINLPALRKLDSMLMEALDSFQN-TEFWYAEEG--SLSMKSARS--STGSFRKVIVQRKEE 260 AT2G45890_AtRopGEF4_ GQSLRLQPLSTEKKDLWKREMNCFMSICDYIVEVIPR----S-LGTNVEITETKLRSDILMSLPALRKLDNMLMEILDSFTE-NEFWYVERG--SSSMNSGGGGRDSGTFRKVVVQRKDE 247 LOC_Os02g47420_OsRopGEF4_ GELWKLEPLCEEKKVRWRKEMDWLLSPTTYMVELVPTKQNGA-DGCTFEIMTPKARSDVNVNLPALQKLDSMLIEVLDSMVD-TEYWYVESG---SRANGRGKKNGLRQTK 273 AT1G31650_AtRopGEF14_ GELWKLEPLCEEKKQKWRREMDWLLSPTNYMIELVPSKQNDA-NGRSLEIMTPKARADIHMNLPALQKLDSMLIETLDSMVN-TEFWYSEIG---SRAEGKNKST--SESK 282
* :*:.: : * :*: ::. :.:. . * :: * *: :*:*.*:: :*: : : :: * . 62
370 380 390 400 410 420 430 440 450 460 470 480 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ KWWLPPVKVPPGGLSEPSRRMLYFQKDSVTQVQKAAMAINAQVLSEMEIPESYIDSLPKNGRASLGDSIYKSIT-EEWFDPEQFLAMLDMSTEHKVLDLKNRIEASVVIWKRKLHTK--D349 AT4G13240_AtRopGEF9_ KWWLPPVKVPPNGLSESARRMLHFQKDSVSQVQKAAMAINAQVLSEMAIPDSYIESLPKNGRVSLGDSLYKSIT-EEWFDPEQFLSTLDLSTEHKVLDVKNRIEASIVIWKRKLHLK--D338 AT5G19560_AtRopGEF10_ NWWLPVVKVPTDGLSEESRRWLQNQKDSVAQVLKAATAINAHVLSEMHVPENYIDSLPKNGKTSLGDFLYKSIT-EESFDPDYFVSFLDLSTEHKVLDLKNRIEASMVIWKRKMCQKEKD310 LOC_Os02g17240_OsRopGEF3_ KWWMPTVKVPAEGLSEVTRKWLQYQKECVNQVLKAAMAINAQVLVEMEIPEVYIESLPKKGKTSLGDAIYRSIT-DEEFDPIEFLEGVDLSTEHKVLDLKNRIEASTIIWKRKMQTK-QE330 LOC_Os05g48640_OsRopGEF2_ RWWLPTVRVPPGGLSDASRKWVQHQKDLVNQVLKAAMAINANVLMEMDVPEAYIESLPKNGKSTLGDSMYKIIT-EDHFNPEELLGTVDMSAEYNIIDLKNRIEASVVIWQRKMVHK--E378 LOC_Os01g48410_OsRopGEF6_ KWWIPTVRVPPEGLPDASKKWILHQKDLVGQVLKAAMAINADVLTEMEIPGEYIETLPKNGRSSLGDSIYKIIT-DDHFDPNELLSSVDLSTEHKIVDLKDRIEASVVIWQRKISN----382
LOC_Os01g55520_OsRopGEF8_ KWWIPIVKVPPGGLSPASRGWLQHQKELVNQVLKAAMAINANCLMEMAIPESYLESLPKNGRASLGDALYRIIT-DVEFDPDVFLSTVDLTSEHKILDLKDRIEASVIIWNRKVHNK--D335 AT1G79860_AtRopGEF12_ KWWIPTVKVPPDGLSEASRRFLQYQKDCVNQVLKAAMAINAQVLFEMEIPESYIDSLPKNGRASLGDQMYKNIT-VDFFDPDQFLSSMDMSSEHKIVDLKNRIEASIIIWKRKMVYKDNK355 AT1G52240_AtRopGEF11_ KWWLPAVKVPPNGLSEISRRFLQSQKECVNQVLKAAMAINAQVLSEMEIPESYLESLPKNGRASLGDVIYRMIT-VEMFDADQFLIEMDLSSEHKILDLKNRIEASIVIWKRKMVQKDTK431 AT3G16130_AtRopGEF13_ KWWLPIVKVPPKGLSETLKRFLLSQRECVCQVLNSAMAINSQVLTEMEIPESYIDSLPKKGRASLGDMIYRMIT-LEMFDAEQFLLEMDLSSEHKILDLKNKFEASVVIWQRKIVQIDNK394 LOC_Os01g62990_OsRopGEF5_ KWWLPVPRVPPGGLRDKARKQLQHKRDCANQILKAALAINSNALAEMEVPESYLESLPKNGRATLGDIIYRYIT-SDHFSPECLLDCLDLSTEYQALEIANRVEASVYVWRRRIAAKPAS346 LOC_Os05g38000_OsRopGEF10_ KWWLPVPRVPPGGVGETTRRQLEHKRDCASQILKAAMAINSNALAEMDVPDSYLDSLPKNGRATLGDIIYRYIT-SDQFSPDCLLDCLDLSSEYQALEIANRVEASIYVWRRRGTSGAAS377 AT5G02010_AtRopGEF7_ KWWLPVPRVSPGGLQENSRKQLQHKRDCTNQILKAAMAINSITLADMEIPESYLESLPRKGRSCLGDLIYRYIS-SDQFSPECLLDCLDLSSEHQAIEIANRVESSIYLWHKRTNSKPAT344 AT3G55660_AtRopGEF6_ KWWLPLPRVPSDGLTEQTRKKLDHTREFTNQILKACMSINSIALAEMEVPQSYLEALPKNGRSCLGDFLYRNIT-SDNFSADHLLESIDLSSELAVVEMANRVEASMYVWRRRAHSRHLI375 AT5G05940_AtRopGEF5_ KWWLPVPRLAPNGLTEEARTELNHKRECATQILKAAMAINSLALTEMDVPKSYLETLPKNGRSCLGDVIYRYVT-SDKFSAESLLDCLDLSSEHIALDIANRVEASIYVWRRRVQTKLGV374 LOC_Os09g37270_OsRopGEF7_ KWWLPCPRVPPKGLSEDARRKLQQDRDCANQILKAAMAINSDVLAEMEIPEVYLESLPKSGKSCLGEIIYRYIT-AEQFSPECLLDCLDLSSEHHTLEVANRIEAAIHVWRLKGQKKSTP363 AT4G38430_AtRopGEF1_ KWWLPCPKVPPNGLSEEARKKLQQCRDFANQILKAALAINSGVLAEMEIPDPYLETLPKSGKECLGEIIYQYLT-ANKFSPECLLDCLDLSSEHQTLEIANRIEAAVHVWRQKNGRRHKK362 LOC_Os10g40270_OsRopGEF1_ RWWLPVPCVPDGGISGKARKELQQKRDCATQIHKAAVAINTGVLGDMEVPDSFMAILPKSGKASVGDGVYRAMLGAEKFSPEYLLDCLDMSSEHEALAMADRVEAAMYVWRRKA-GASHG389 LOC_Os04g47170_OsRopGEF9_ KWWLPVPCVTKPGLTESARRDLRQKHDCASQIHKAAMAINNGVLAEIRIPELYKQTLPKCGRASVGDLIYRHMSFPGKFSPEYLLDRLEISSEHDALEAADRVEAAMHVWRRKA-SQGHS368 LOC_Os07g29780_OsRopGEF11_ KWWLPEPCVPESGLSDRMHRELQQKRDQASQIHKMAMEINSGILSEMQVPLSYIETLPKSGKVGVGDAIYRYMSAGDQFSPDHLLNFLNLSSEHEALEIADRVEAAMYVWRRKA-SMTHV352 AT1G01700_AtRopGEF2_ KWWLPIPLVPLQGLSEKARKQLKSKRESTNQIHKAAMAINSSILGEMDIPDSYMATLPKSGKASTGDAIYRHMTSSGRFSPEKLLDRLKIVSEHEALQLADRVEASMYTWRRKA-CLNNS391 AT4G00460_AtRopGEF3_ KWWLPVPLVPSEGLSDKARKQLKNKRESTNQIHKAAMAINSSILSEMEIPDSYMTTLPKCGKSSVGDSIYRYMSGSGRFFPEQLLDCLNISSEHEAVQLADRVEASMYTWRRKS-CLSNS379 AT2G45890_AtRopGEF4_ KWWLPVPCVPAEGLSEEERKHLRHKRDCASQIHKAALAINDSTLNDMDIPDSYLTTLPKSGKASVGDVIYKQLCTAEKFYPDQLLDILKITSEHEALELADKVEASLVTWRRKTGGLTHS367 LOC_Os02g47420_OsRopGEF4_ KWWLPSPRVPDIGLSQFQRNRLVFQAKLVHQILKAAMSINEEVLLQIPIPPAVTDALPKSGRAGLGEDLYHAIT-TEYIPIEEIFLSLSLKTEHTVLETMNRLEGAVFAWNQRISEEKSK392 AT1G31650_AtRopGEF14_ RWWLPSPQVPKPGLSNSGRKKLLDKGKVVYQVFKATKAINENILLEMPVPIVIKEAIPKSGKNSLGDELYKMLA-VESATVDEIFISLNLGTEHAALETVNKLESAMFAWKERITEQGSN401 .**:* :. *: : : . . *: : ** * :: :* :*: *: *: :*: : :. :.: :* : ::.*.: *. : 104
490 500 510 520 530 540 550 560 570 580 590 600 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ TKS---SWGSAVSLEKREL---FEERAETILVLLKQKFPGLPQSSLDISKIQFNKDVGQAVLESYSRILESLAYTVMSRIEDVLYTDTLALKQTLLAEET---443
AT4G13240_AtRopGEF9_ NKS---SWGSAVSLEKREL---FEERAETILVLLKQKFPGLPQSSLDISKIQYNKDVGHAVLESYSRILESLGYTEMSRIDDVLYADSLARKQ-CTGEET---431
AT5G19560_AtRopGEF10_ GKS---QWGSTVSLEKREL---FEVRAETILVMLKQQFPGIPQSSLEVSKIKNNKDVGQAILESYSRVLESLASKIMSRIEDVLEADRLVQRQLMGEAETRS---406
LOC_Os02g17240_OsRopGEF3_ AKS---SWGSIISFEKREQ---FEERAETILHLLKLQFPGTPQSQLDISKIQYNRDVGYAILESYSRVLESLAYSVMSRIEDVLGADAAAQNLTATEAARR---425
LOC_Os05g48640_OsRopGEF2_ GKL---SWGHGVKFEKREK---FEARAENVLLLIKHRFPGIAQSALDISKIQYNRDIGLAILESYSRTLESLAFTVMSRIEDVLHADSLAQASNTRTQESMRM---475
LOC_Os01g48410_OsRopGEF6_ -KL---SWGPGVSLEKREQ---FEERAQTVLLILKHQFPGVPQSSLDISKIQYNKDVGYAILESYSRTLESLAFAVLSRIEDVLHADAIARDP--KRTKSRRR---476
LOC_Os01g55520_OsRopGEF8_ GKS---AWGSAVSQEKREQ---FEERAQTLLLIIKHRYPGIPQSTLDIAKIQENRDVGFAILESYSRVLESLAFNVMSRIEDVLNADDHAREKAKKEAPPAP---431
AT1G79860_AtRopGEF12_ SSA---PWASGVSLEKREV---FEERAETILLILKQRYPGISQSSLDISKIQFNEDVGQAVLESYSRILESLAYTVLSRIDDVLEADR---AGNKRN---443
AT1G52240_AtRopGEF11_ S---PWGSTVSIEKREQ---FEERAETILLLLKQGFPGISQSSLDISKIQFNRDVGLAILESYSRVLESLAHTVMSRIEDVLYADQLTQEPTNNAPSKNR---525
AT3G16130_AtRopGEF13_ SSS---PWSTNLSMDKRQQ---LEERAATILQLIKQEFPGISQSTLDISKIQFNRDIGLAIVESYSRILESLAHTVMSRIEDVLEADQLTQNP-ELAMCKIH---489
LOC_Os01g62990_OsRopGEF5_ VLGRATSG--RSSWGMVKDMIIDTE---KRE-LLAERAEGLLICLKQRFPGLTQTSLDMSKIQYNRDVGKSILESYSRVLESLASNIVARIDDLLNIDELNRH-AEHFP---448
LOC_Os05g38000_OsRopGEF10_ RAG----N--KSSWGIVKDMIMDTE---KRDDLLADRAEGLLMCLKQRFPGLTQTSLDTSKIQYNKDVGKSILESYSRVLESLASNIIARIDDLLYVDERSRQ-AELLPT---AG479 AT5G02010_AtRopGEF7_ NTK---TSWEMVKELMVDAD---KLE-LMADRAESLLLSLKQRFPGLPQTALDMSKIQYNKDIGKSILESYSRVLESLAFNIVARIDDLLFVDDLTRHSSDQIPT---TL444 AT3G55660_AtRopGEF6_ SLYRSTSTRWGMIVKEMMMHQTDGD---KRE-IFAERAESLLIRLKQRFPGLRQTALDTSKIQYNKDVGKSILESYSRVLESLAYSIGVRIEEVLFMDDISKDDGDGDDDSCSDKLRLLS491 AT5G05940_AtRopGEF5_ NNN-TSSTTPKLTWEMVKELMAAGD---KRG-LLVERSETLLRCLKQRFPSLTQTSLDISKIQWNKDIGKSILESYSRALESLASNIIARIDDLLYVDDLTKQSDDNN---LLS480 LOC_Os09g37270_OsRopGEF7_ QAK----SKKS-WGGKVKGLVGDTE---KSH-VLSQRADGLLQSLRLRYPGLPQTSLDMNKIQYNKDVGQSILESYSRVLESLAFNIIARIDDVIYVDDATKKS-AAADS---463
AT4G38430_AtRopGEF1_ QAK----LKLSSWGGKVKGLVNDNE---RND-FLVQRAETLLQSLRIRFPGLPQTTLDMNKIQYNKDVGQSILESYSRVMESMAFNITARIDDVLYVDDAMRRSISVTES---464
LOC_Os10g40270_OsRopGEF1_ RSR---WGAVKELVADDEE-QDKNVMLAGRAESLLLCLKHRFPGLSQTTLDTSKIQFNKDIGQAILESYSRVLESLAFNIVSWIDDVLFADKTVRKQ---482
LOC_Os04g47170_OsRopGEF9_ RSP---WSAVKELMES---DKNVMLASRAGDVLLCLKQRFPGLSQTTLDASKIQYNKDVGQAILEGYSRVLESLAYNIVTCIDDVLFADESARKI---457
LOC_Os07g29780_OsRopGEF11_ VSK---WENVTELNADG----DKNLILASRARSLLLCLKQRFPGLSQTTLDTSKIQYNKDIGQAILESYSRVLESLAYNIVSWIDDVLIADGNARKGHNIRMQ---448
AT1G01700_AtRopGEF2_ KSS---WNMVKDLMSITER-SDKNYVLAERAESLLFCLKQRYPELSQTSLDICKIHCNKDVGKAVLESYSRVLEGLAFNIVAWIDDVLYVDKTMRGEE---485
AT4G00460_AtRopGEF3_ KNS---WNMVKDLMSTTER-TDKNYVMAERAETLLFCLKQRYPELSQTSLDICKIQYNKDVGKAVLESYSRVLEGLAFNIVAWIDDVLYVDKTMRGSE---473
AT2G45890_AtRopGEF4_ KSS---WDMMKDISGDADRGNDKNHILAARARSLLFCLKQRYPELSQTSLDICKIQFNRDVGKAVLESYSRVLEGLAYNVVSWIDDVLYVDRTVRNRDD---463
LOC_Os02g47420_OsRopGEF4_ -KSPGR---HSWNFMKDSSSELD----KMSMCIERVETLMQLLKSRFPSLPPTFIEVVKIQYNVDVGHAIVEAYSRVLVGVAFSILSRVAEILLEDDLIKKPNTPLATLKFD---496
AT1G31650_AtRopGEF14_ GKSPVR---ASWSFAKDPLSEIG----RNESLLNRAEALRTQIKSKHPNLPHSFLDATKIQYDKDIGHAVLEAYSRTLANLAFRILSRMGEILKEDSLSN-PNSPAPPSCFP---505
* : :: .* : :: **: : *:* :::*.*** : .:. : ::: * 610 620 630 640 650 660 670 680 690 700 710 720 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ ---SDGG-RTTETDSESAGSSN-SGEEAEKHDPHS---KTLLDFMGWNDNSSKG-GDKPT---KSPNLTP---KKLSYLEKLENLNGFRSPKD-RH---523
AT4G13240_AtRopGEF9_ ---SDGGKIATETDSASAGSSNYSGEEIEKLESQNSSKTTLLDFIGWSDNSSKGQSEKPP---KSPRMTP---KKLSYLEKLENLNGFRSPKD-RH---517
AT5G19560_AtRopGEF10_ ----ESEAESEYEETEKVVAAETPNSRKLSDFIGWRLSSDTKKHSSMSDIEFFHKVEQEKEKP--MMKSPRALP---KKFSYLAKLENM---RSPSD-RH---493
10 20 30 40 50 60 70 80 90 100 110 120 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ ---MVAAL---ERGLSASKS---FNFKRMFDSSSTKQQQSQTIVVENGDSHIVES46 AT4G13240_AtRopGEF9_ ---MVPSL---ERGISITSS---FNLDRMFDSSPGKEQQ---QPHLAET37 AT5G19560_AtRopGEF10_ ---MFDGRNS---GHSSFSSR15 LOC_Os02g17240_OsRopGEF3_ ---MVRFL---RRHSLDKNS---AQFHRQLSHDSS---SSSSDTA33 LOC_Os05g48640_OsRopGEF2_ ---MARPL---LKIGHGLDR---FRWRRSTSSSSSSPLALSLSSSSAAALSDDDP46 LOC_Os01g48410_OsRopGEF6_ ---MVRRH---LLRGHSLDR---FLPIRSLIMSSSSSFSSS-SPSPPPPSSSSSR45 LOC_Os01g55520_OsRopGEF8_ ---MAASGGGS---LLERRSSVRRSQS---MVSEEGR28 AT1G79860_AtRopGEF12_ ---MVRAS---EQEQETYRS---RLFNFK---WRNNDN-NSATRHNKSLSVETGLDEAATG--SHDA52 AT1G52240_AtRopGEF11_ ----MLEGKAMVEDSDMPVKMQMQAMAFASQALDLFDVFDCKSIAGHIKKEFDERYGSGWQCVALLLLNYTTLIMGMPKLREEVDFNDNMNENSASRHVKSWSSDCAMRMDGSDNLDDDD116 AT3G16130_AtRopGEF13_ ---MVKAS---EKEHEKYKP---KLFDLE---NVKKKNSSSRHFKRWNSDSALRIEDPDIDDGTV53 LOC_Os01g62990_OsRopGEF5_ ---MGSG---EEERE---ARSEAAFTDSADGS----SSSSDAASADEWPV37 LOC_Os05g38000_OsRopGEF10_ ---MARGGGG---GEEAAEEERE---VSEALTADSSADEECRRGSSSSSASSGDASSE49 AT5G02010_AtRopGEF7_ ---MDGS---SENLPE---VEEKGRESSCCSSET---TRQEEEEQSPSCTE39 AT3G55660_AtRopGEF6_ MED--NSCIGFEGSKRF---GETSKRIIGL---IDSVTESTTDSSLSSSSSGVGSSSGRSSVAER57 AT5G05940_AtRopGEF5_ MENLVKSCAGIE---KKRSNL---TPSMEDILTESKERSSTSGASYESS-STTTVA49 LOC_Os09g37270_OsRopGEF7_ ---MASAS---EDDAGS---ERCCGSYSPSADVSESETSSDCSAPTTTTTTR43 AT4G38430_AtRopGEF1_ ---MGSLSS---EEDDEVS---SERCGSYSPSADISESESSSSFSCHRFDGEGA45 LOC_Os10g40270_OsRopGEF1_ ---MESSSSV---SVSDESSEANGGGCCS---SSTTPSMDAVNLSRTFSDVSSFSEEHGGSGSSVDH58 LOC_Os04g47170_OsRopGEF9_ ---MDTPS---TTCDESSEVDAR---DDYGDIDDVERRR---GRHRREASSDVSSEC45 LOC_Os07g29780_OsRopGEF11_ ---MSGGSGF---PGFHKHG---YDRDYARPLFR---VSSFSDNGGGEEQEH40 AT1G01700_AtRopGEF2_ ---MENLPNH---EENDDVG---YHQSPGPIDPNDH-SASETPVYSTMSTDSFAYH46 AT4G00460_AtRopGEF3_ ---MENLSNP---DENDD---HQSPRSIDQNDQ-SAVETPVYSTMSIDSFVYP43 AT2G45890_AtRopGEF4_ ---MESSSNS---DQNEG---TPTSSVSSPYRR-TYSDISGLSHR-FDVQSFY42 LOC_Os02g47420_OsRopGEF4_ ---MRMKTLACCRRRPQ---DFSIDMDQ-EPDRVMTYNG---LESCIINSSSYDDDSGLSATTGADGCVTTDSV64 AT1G31650_AtRopGEF14_ ---MMLMRRRLACCSRER---EISIDFDEHEQDKMITYYG---LETCIINNQSYEEESG---TSRGDGCLTDSLD63 130 140 150 160 170 180 190 200 210 220 230 240 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ NTPE---SQNSDSFVESPVE---SSLPMISPLTRPGKRSERQQADMEMMKDRFAKLLLGEDMSGGGKGVSSALALSNAITNLAA---SIF127 AT4G13240_AtRopGEF9_ TMPE---SQTQDSLGGSPVE---TSRPMTSRLIS--RRQDKQQSETEMMKDRFTKLLLGEDMSGGGKGVSSALALSNAITNLAA---SIF116 AT5G19560_AtRopGEF10_ GDGM---HTPEHELAGHAAP---STR---RGKQNRRSDMEVMKERFAKLLLGEDMSGGGTGETSALALSNAITKLAD---SMF86 LOC_Os02g17240_OsRopGEF3_ DMHE---PHGNGSGGTPPLP---NGRAAAAGAPRSRFGRDGPPSDLDVMKERFAKLLLGEDMSGTGKGVSSALALSNAITNLAA---SVF114 LOC_Os05g48640_OsRopGEF2_ GSPMDPEMPPAARRALSRSSGSRGRLSFELPPLAG---GPSDKEEAPPR---TSSAPAPARPAPAALHE-GPPSDAEMVREKFSKLLLGEDMSGTGKGVSSALALSNAITNLAA---SVF156 LOC_Os01g48410_OsRopGEF6_ GSSSGRWCGVSVAEEDEDDAAVSATTTPPLPPLQKRVLSRSHGSKAKPLGGSVDHLPPVVPPSSKNVRDSGPPSEIDLVKEKFAKLLLGEDMSGTGKGVSSALALSNAITNLAA---SVF162 LOC_Os01g55520_OsRopGEF8_ GTPA---DEDLGGGGTLKIGAVLDK---DSAAPKSRLAKDTGEHGGGGPSEMELMKEKFAKLLLGEDMSGSGKGVPSALAVSNAITNLAA---SVF115 AT1G79860_AtRopGEF12_ EPLTIIHPSQG---PPLSR---SAADEA---VLAALAASQARERQ--LLADMEQMKERFSKLLLGEDNSGGGKGVSSALALSNAITNLAA---SVF134 AT1G52240_AtRopGEF11_ NDMMMFRSQPGKCGSVDRPSLPIGGVTPNRND---KLPRVSSSDSMEALIILQAAMEQMKEKFSKLLLGEDMSGGGKGVSSALALSNAITNLAA---SAF210 AT3G16130_AtRopGEF13_ FKKTATSSIQPILPVLAMDEQPQPREATDEESQKDSGKREIISFLGYNEKFTRDQVILRFRGLQAKSCYFAYVTELEQMKDKFAKLLLGEDMSGGSKGVSSALALSNAITNLAA---SAF170 LOC_Os01g62990_OsRopGEF5_ TLAAPP--RKTAACGRVPGAEVVDSSKPHAQKRRAPS---SEMEMMKERFAKLLLGEDMSGSGKGVCTALAIANAITNLCA---TIF116 LOC_Os05g38000_OsRopGEF10_ SYCPPDEWQKVAIKTCVSSDLVVVSAEPAKEKKPPPPPSSP---RVDAAPADKHHRPSEMEMMKERFAKLLLGEDMSGGGKGVCTALAISNAITNLCA---TIF147 AT5G02010_AtRopGEF7_ DFTASP---VSSRWSVKNIDGEKKKIRSDS---RVSEVEMMKERFSKLLLGEDMSGSGNGVCTALAISNAITNLCA---TLF112 AT3G55660_AtRopGEF6_ SVSSPPTKSQILGWPLGQGSWRKSSGK---MKKKTPTKID---DFGFKRVGTETSEIELLKERMAKLLLGEDMSGSGEGVCPALAISNAITNLYA---AIL149 AT5G05940_AtRopGEF5_ SSSPPPPPSQILGWPIRKASFRKNSKESVNFDHKKLTLHD---DSGFKAKEMNSADVEMMKERFAKLLLGEDMSGSGKGVCTALAISNAITNLCA---TIF144 LOC_Os09g37270_OsRopGEF7_ RFASSSSRGVASSS---SSSLLPTPPPSSAAAFFLSAKPA---ADLSEVDMMKERFAKLLLGEDMSGSGKGVCTALAISNAITNLSA---TVF127 AT4G38430_AtRopGEF1_ SSSIPSSPRVVAGRGFYFPAPVMLPVIGGKDVVWDDKQPD---NDLSEIEMMKERFAKLLLGEDMSGGGKGVCTALAISNAITNLSA---TVF132 LOC_Os10g40270_OsRopGEF1_ SGPFEPPSAAAVSRLIGRRSPAAAASALSLSRLSMKPRADV---LDRRST-DDELELVKERFSKLLLGEDMSGGGKGVCTAVAISNAITNLYANAATVF153 LOC_Os04g47170_OsRopGEF9_ SGEPGSPYGSPYPRWP-VCSIAKAPPPPLLQKLGAARRGAG---RDRKAG-DGELQLIKERFSKLLLGEDMSGSGKGVSTAVAISNAITNLYA---TVF136 LOC_Os07g29780_OsRopGEF11_ YTP-SPPRGRSMSRTT---STAPRLSP-SVSKLSMKKLQQV---VNEKSLEDEEMELMKEKYTKLLLGEDMSGSGKGVCTAVAISNAITNLYA---TVF128 AT1G01700_AtRopGEF2_ RTCSETSGGGFSDQIDETSSFCTEASPSDWPVLTESNNSASSNFPTVFD---LKHNQIETDEHLAVQEISEPELETMKERFSKLLLGEDMSGSGKGVCTAVTISNAITNLYA---TVF158 AT4G00460_AtRopGEF3_ RTCSETTSG-FSDQIDETNSFCSEASPCNWPVLTESKSSKC---LSGLEMQSNECLVVQEISEPELETMKERFAKLLLGEDMSGSGKGVCTAVTISNAITNLYA---TVF146 AT2G45890_AtRopGEF4_ NRPSNTNAVVLSGHEEDVSEDAEEPKD---DVVNDVHGDGD---EEDSD----IDSAEDAELEMMRERFAKLLLGEDMSGSGKGVCTAVTVSNSITNLYA---TVF135 LOC_Os02g47420_OsRopGEF4_ DDEVSSCSSSKDVSSSSFSSQCHPLRKQEEHSLYELDTLSAVHLLPLKGKKPITYTLS---ASDIETMKEKFGKLLLGDDASGGARGVCAALALSNAIINLSA---TIF167 AT1G31650_AtRopGEF14_ DDAFSSCSSSKDASSS-FSSKWLPMKN-DEHSCDGLNLSGRSQHFDAKEKKKQGYGSSQHFDAKEKPGYVYCHLDVEAMKEKFSKLLLGEDVTGGCKGVQVALALSNAVTHLAT---SIF178 : :::: *****:* :* * *::::*:: :* : : 250 260 270 280 290 300 310 320 330 340 350 360 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ GEQTKLQPMPQDRQARWKKEIDWLLSVTDHIVEFVPSQQTSK-DGVCTEIMVTRQRGDLLMNIPALRKLDAMLIDTLDNFRGHNEFWYVSRD---SEEGQQARNDRTND 232 AT4G13240_AtRopGEF9_ GEQTKLQPMAPDRRARWKKEIDWLLSVTDHIVEFVPSQQISK-EGVCTEIMVTRQRGDLLMNIPALRKLDAMLIDTLDNFRGHNEFWYVSRD---SEEGKQARNERTKD 221 AT5G19560_AtRopGEF10_ GEQMKLQPMYPETKENWRKEMGWLLSVIDHIVQFVPSRQMGK-NGQFTEIMVTKQRDDLLTNIPALRKLDSVLLETLDNFKDQKDFWYVPRD---MEDADHNGDWRRDE 191 LOC_Os02g17240_OsRopGEF3_ GEHRKLEPMAPDTKERWKKEVGWLLSVTDHIVEFVPTRQTAE-NGTTMEIMSTAQRRDLQMNVPALRKLDAMLIGYMDNFVDQSEFWYE---KGGDNKRDDD 212 LOC_Os05g48640_OsRopGEF2_ GEQRRLQPMAADQKARWRREIDWLLSVSDHIVEFVPSKQVSE-DGSTMEIMITQQRRDLQMNIPALRKLDAMLLEYLDNFKDKQEFWYVSKD---ASESEKGNTPRQDD 261 LOC_Os01g48410_OsRopGEF6_ GEQRRLEPMSADRRARWNKEIDWLLSVTDHIVEFVPSQQAST-DGTSMEVMGTQQRRDLLINIPALRKLDAMLLEYLDNFKDEQEFWYVKKD---ADEGEKGDAPRQGD 267 LOC_Os01g55520_OsRopGEF8_ GEQRKLEPMAPDRKGRWKKEVGWLLSVADHIVEFVAKKQVLD-NGVEMEVMGTQQRRDLQANIPALRKIDTMLLDYLDNFKDRNEFWYVKRD---S--CSDSDEQRSDE 218 AT1G79860_AtRopGEF12_ GEQRRLEPMPAERRARWRKEIDWLLSVTDYVVEFAPSQQKNK-DGTNMEIMTTRQRTDLHMNIPALKKLDAMLIDCLENFKDQSEFSYISKD---SPDL---DGKRNDE 236 AT1G52240_AtRopGEF11_ GEQRRLEPMAVDRKTRWRREIGWLISVADYIVEFAPTQQTNK-DGTSMEVMSTRQRTDLLCNIPALKKLDAMLLDCLDKFKDQDEFYYVKKD---SPDS---CETRNDE 312 AT3G16130_AtRopGEF13_ GEIRRLEAISEDKKERWRREIGWLLSVTDHIVEFSPTHQTNE-DGSSMEVMTTKQRTDLVSNIPSLKKLDEMLLDCLDKFKDQDEFYYVTPG---SPESENSNSTRNDD 275 LOC_Os01g62990_OsRopGEF5_ GQLWRLEPLPPEKKAMWRREMGWLLCVSDHIVELVPTWQSFP-DGTRLEVMTSRPRSDLYINLPALRKLDHMLIEILDSFRD-PEFWYVEQG--ICAPD---CDGSASFRAAFHRRDE 227 LOC_Os05g38000_OsRopGEF10_ GQLWRLEPLLPEKKTMWRREMDWLLCVSDHIVELVPTWQTFP-DGSKLEIMTSRPRSDLYINLPALRKLDHMLLEILESFRD-PEFWYVDQG--ICPPD---RDGSAPFMLTFHRRDE 258 AT5G02010_AtRopGEF7_ GQLWRLEPLPTEKKEMWRREMEWLLCVSDHIVEMTPTWQTFP-DGTKLEIMTCRPRSDLYVNLPALRKLDNMLLEILDSFEE-TEFWYVDQG--IMAHESA---ADGSSSFRKSFQRQED 225 AT3G55660_AtRopGEF6_ GQQWRLEPIPSEKKLMWRREIEVLLSVSDHIVELVPSFQNFP-NGNKIEVMNCRPRSDLFTCLPALRKLDNMLIEILDSFGE-TEFWYVDQG--IVAAE---SARSNSFR----EDGD 256 AT5G05940_AtRopGEF5_ GQLWRLEPLSSEKKEMWRREMEWILSVSDHIVELTPSTQTYP-DGNKFEVMTCRPRFDLFINLPALRKLDNMLLDILASFKK-TEFWYVDQG--IVASE---NDGSASFRRKIQRQEE 255 LOC_Os02g17240_OsRopGEF3_ ----LMDGDAAARKLDAMEELEKLNEAPASMTLYDFMGWHFDQDELMRKKEEGTLDEAGEAK---LKKAPSLAP---KKFSYVDSLSAG-GMRSPSA-RH---513
LOC_Os05g48640_OsRopGEF2_ --ASLSRYDTDKVVIDAKAEVERLGRMEPVSATLFDFVSPRDQDVVATKMDSKEKGCRGDAHSRKLTKVSPIAT---KRYSYLEKLENLSGTRSPIS-RH---569
LOC_Os01g48410_OsRopGEF6_ --PSLV--DIPEIIDNALEEEETVNSIDANSQVTNNSIHWQEQ---EHEDKGAR---LRKVHRMVT---KKLLHIEKVDNLGGGLKSFS-HR---554
LOC_Os01g55520_OsRopGEF8_ ----AMANDAAEHHHQQAGEVDAPCKMTGSPNGRTLLDFMDDWNGDADRPSPTAPEPAAQEDGR-LMKLPNIMTN---LKQTYMDNLFGA--HRSPPG-RH---521
AT1G79860_AtRopGEF12_ ---TPLEAE--EETLVGSMTLSDFMG---WDFDQAANAELESKKD----LPDDPLIKEKLS-VVTT---KKTSYLETLGGV---KSPTA-RH---515
AT1G52240_AtRopGEF11_ ---YSLKEN--EKLREERLSFTEDMASG---TLSDVMQWGNKNNEMKKESFFGDREKPLLSKVTG-IMTNN---KKSSYLDNLGAM---RSPTA-RYS---607
AT3G16130_AtRopGEF13_ ---IVKETESPEKEEEPNFCLLEDRPKKQKPTISLSEVMQWNIETNEPRKE----KSDKKLLTRVSSMIMSNN---KKTTYLESLGTT---RSPTAGRYS---576
LOC_Os01g62990_OsRopGEF5_ --QGDADCRIACNKAAVPPYQVP-ASGTP--FVT---AYATPSFSPAQLASP-SKKERSPLGAGRRSYSNRGFGAKKALAIDLVNPEVMGVIISGGKMIDVSTTTEL---546
LOC_Os05g38000_OsRopGEF10_ AGSGKISCMPAMSASSVPAYPVVSTSGTPPPYAT---AYATPSFSPAQLSSP-SNIGRALL-VDRRSHDGRAFDG---SMEFMGMAVSN-AVFDLPGL---568
AT5G02010_AtRopGEF7_ GNNGND--APKSIAVPVSNYTTPSYSPSKQELRS---SITVPP-SPSRFKIPHSSSVKRVLTAYVTKNEPRLKNLPLERSSRSSSSERLSLEKCMKESLNVSNLDPGI---546
AT3G55660_AtRopGEF6_ KEAASGGSGSLREKLSAPSLFSVSFSGTSTPYRTLSFSASTPSYSPMPLISP-INGGRGGERAPFLSG--RNIRER----CGFGPKKALANYLRG---579
AT5G05940_AtRopGEF5_ SPAVSS---IIAHKKVVPLPYLISASG--TPYRT-SFST-TPGFSPS-MISP-KKGER---RTPYSSKDTNKIIEKGLPSRGYGVRRVLNNYLGMESKLKICVNPSDNADTAVINQISKD588 LOC_Os09g37270_OsRopGEF7_ VSIFNRGIGVPVQKRISPSPFSIQHTPYASPFATPTFCSSTPVTGSPGRVQPPLNKDN--LPTKQEVKVEKLFSG---DIEKVWTYAGNLSARKDAGDAPERD---561
AT4G38430_AtRopGEF1_ LSLFSIN-GLNPQK---AFSVQSSPHGSPFATPALSVAS---RSP-RRAPPLYS---VKRNGTREKGIVG---ETEKAWSYAGNLSSRRVTGVTPERD---548
LOC_Os10g40270_OsRopGEF1_ ---482
LOC_Os04g47170_OsRopGEF9_ ---457
LOC_Os07g29780_OsRopGEF11_ ---KQEFSKLSPQQY---460
AT1G01700_AtRopGEF2_ ---485
AT4G00460_AtRopGEF3_ ---473
AT2G45890_AtRopGEF4_ ---463
LOC_Os02g47420_OsRopGEF4_ ----LSSDVYLAGITETPPGHIRRSLMDQISLVDG---SLDAVVRKKGVKQLRW---543
AT1G31650_AtRopGEF14_ ----SSRDPYRTPERPLLSSRVRHSLTDDMNKADGTETGLDFLFADAKASNSVNTTPSRSSRLWCLSKVPSDTSP---576
730 740 . . . . | . . . . | . . . . | . . . . | . . . AT3G24620_AtRopGEF8_ ---523
AT4G13240_AtRopGEF9_ ---517
AT5G19560_AtRopGEF10_ ---493
LOC_Os02g17240_OsRopGEF3_ ---513
LOC_Os05g48640_OsRopGEF2_ ---569
LOC_Os01g48410_OsRopGEF6_ ---554
LOC_Os01g55520_OsRopGEF8_ ---521
AT1G79860_AtRopGEF12_ ---515
AT1G52240_AtRopGEF11_ ---607
AT3G16130_AtRopGEF13_ ---576
LOC_Os01g62990_OsRopGEF5_ ---546
LOC_Os05g38000_OsRopGEF10_ ---568
AT5G02010_AtRopGEF7_ ---546
AT3G55660_AtRopGEF6_ ---579
AT5G05940_AtRopGEF5_ VEEEKKRNSTSVHQKGPPKYTVS611 LOC_Os09g37270_OsRopGEF7_ ---561
AT4G38430_AtRopGEF1_ ---548
LOC_Os10g40270_OsRopGEF1_ ---482
LOC_Os04g47170_OsRopGEF9_ ---457
LOC_Os07g29780_OsRopGEF11_ ---460
AT1G01700_AtRopGEF2_ ---485
AT4G00460_AtRopGEF3_ ---473
AT2G45890_AtRopGEF4_ ---463
LOC_Os02g47420_OsRopGEF4_ ---543
AT1G31650_AtRopGEF14_ ---576
C1
T 458 S 500 S 510
WW-motif
WW-motif
Supplementary Fig. S3
C2
C3
4
Additional file 3: Figure S3. Protein sequence alignment domain analysis and conserved phosphorylated amino acid residues of C-termini of RopGEF genes. Every OsRopGEF and AtRopGEF protein sequence was collected and aligned, followed by the PRONE domain (C1, C2, C3) and the WW-motif. At the end part of the sequence, we found some conserved regions.
According to previous studies, S510 in the C-terminus of AtRopGEF12 is involved in the C- terminal inhibition of GEF activity. As in AtRopGEF12, the serine residue is conserved in each
LOC_Os09g37270_OsRopGEF7_ GELWRLEPMASARKAMWTREMDWLLSVADSIVELTPSIQELPDGGGQFEVMVPRPRSDLYMNLPALKKLDAMLLAMIDGFKE-TEFWYVDRG--IVVDDSGGPFSSSSSSCGRPSVRQEE 244 AT4G38430_AtRopGEF1_ GELWRLEPLAPQKKAMWRRELEWLLCVSDSIVELIPSIQQFP-GGGTYEIMETRPRSDLYANLPALKKLDAMLIDMLDAFSD-TEFWYTDRG--IVLGDCD---KDSYNSPASVRQED 243 LOC_Os10g40270_OsRopGEF1_ GSCHKLEPLPAGKKTMWRREMDCLLSVCDYIVEFYPSSQTLP-DGTKVEVMATRPRSDIYINLPALEKLDAMLIEILESFQK-AEFWYADAGTRSFGSATSSSTMSSSSFRRSTH-RNED 270 LOC_Os04g47170_OsRopGEF9_ GGCHRLEPLLAEKRSMWRREMDCLLSVCDYIVELFPSKEIMP-DGTVREVMATRPRSDIYVNLPALEKLDDMLLEILDSFQK-TEFWYVND----KGQKDSCAAAAAAPCRPVSH-RDGD 249 LOC_Os07g29780_OsRopGEF11_ GTCHRLQPLPPEKKAMWNREMDCLLSICEYIVEFSPKVQAMP-DGSTHDVMATSPRSDILMNLPALEKLETMLLEILDSFEK-TDFWYIDQ---RKQSFSDSMKFQ---RSED 233 AT1G01700_AtRopGEF2_ GQNLRLEPLEIEQKTTWKREMNCLLSVCDYIFEFIPKSQNLS-NGATVEVMESRPRADIYINLPALRKLDSMLMEALDSFQK-TEFWYAEEG--SLSMKSTRS--ATGSFRKVIVQRKEE 272 AT4G00460_AtRopGEF3_ GQNLRLEPLETEKRALWKREMNCLLSVCDYIVEFIPRCQNLS-NGATVEVMESRPRADIYINLPALRKLDSMLMEALDSFQN-TEFWYAEEG--SLSMKSARS--STGSFRKVIVQRKEE 260 AT2G45890_AtRopGEF4_ GQSLRLQPLSTEKKDLWKREMNCFMSICDYIVEVIPR----S-LGTNVEITETKLRSDILMSLPALRKLDNMLMEILDSFTE-NEFWYVERG--SSSMNSGGGGRDSGTFRKVVVQRKDE 247 LOC_Os02g47420_OsRopGEF4_ GELWKLEPLCEEKKVRWRKEMDWLLSPTTYMVELVPTKQNGA-DGCTFEIMTPKARSDVNVNLPALQKLDSMLIEVLDSMVD-TEYWYVESG---SRANGRGKKNGLRQTK 273 AT1G31650_AtRopGEF14_ GELWKLEPLCEEKKQKWRREMDWLLSPTNYMIELVPSKQNDA-NGRSLEIMTPKARADIHMNLPALQKLDSMLIETLDSMVN-TEFWYSEIG---SRAEGKNKST--SESK 282
* :*:.: : * :*: ::. :.:. . * :: * *: :*:*.*:: :*: : : :: * . 62
370 380 390 400 410 420 430 440 450 460 470 480 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ KWWLPPVKVPPGGLSEPSRRMLYFQKDSVTQVQKAAMAINAQVLSEMEIPESYIDSLPKNGRASLGDSIYKSIT-EEWFDPEQFLAMLDMSTEHKVLDLKNRIEASVVIWKRKLHTK--D349 AT4G13240_AtRopGEF9_ KWWLPPVKVPPNGLSESARRMLHFQKDSVSQVQKAAMAINAQVLSEMAIPDSYIESLPKNGRVSLGDSLYKSIT-EEWFDPEQFLSTLDLSTEHKVLDVKNRIEASIVIWKRKLHLK--D338 AT5G19560_AtRopGEF10_ NWWLPVVKVPTDGLSEESRRWLQNQKDSVAQVLKAATAINAHVLSEMHVPENYIDSLPKNGKTSLGDFLYKSIT-EESFDPDYFVSFLDLSTEHKVLDLKNRIEASMVIWKRKMCQKEKD310 LOC_Os02g17240_OsRopGEF3_ KWWMPTVKVPAEGLSEVTRKWLQYQKECVNQVLKAAMAINAQVLVEMEIPEVYIESLPKKGKTSLGDAIYRSIT-DEEFDPIEFLEGVDLSTEHKVLDLKNRIEASTIIWKRKMQTK-QE330 LOC_Os05g48640_OsRopGEF2_ RWWLPTVRVPPGGLSDASRKWVQHQKDLVNQVLKAAMAINANVLMEMDVPEAYIESLPKNGKSTLGDSMYKIIT-EDHFNPEELLGTVDMSAEYNIIDLKNRIEASVVIWQRKMVHK--E378 LOC_Os01g48410_OsRopGEF6_ KWWIPTVRVPPEGLPDASKKWILHQKDLVGQVLKAAMAINADVLTEMEIPGEYIETLPKNGRSSLGDSIYKIIT-DDHFDPNELLSSVDLSTEHKIVDLKDRIEASVVIWQRKISN----382
LOC_Os01g55520_OsRopGEF8_ KWWIPIVKVPPGGLSPASRGWLQHQKELVNQVLKAAMAINANCLMEMAIPESYLESLPKNGRASLGDALYRIIT-DVEFDPDVFLSTVDLTSEHKILDLKDRIEASVIIWNRKVHNK--D335 AT1G79860_AtRopGEF12_ KWWIPTVKVPPDGLSEASRRFLQYQKDCVNQVLKAAMAINAQVLFEMEIPESYIDSLPKNGRASLGDQMYKNIT-VDFFDPDQFLSSMDMSSEHKIVDLKNRIEASIIIWKRKMVYKDNK355 AT1G52240_AtRopGEF11_ KWWLPAVKVPPNGLSEISRRFLQSQKECVNQVLKAAMAINAQVLSEMEIPESYLESLPKNGRASLGDVIYRMIT-VEMFDADQFLIEMDLSSEHKILDLKNRIEASIVIWKRKMVQKDTK431 AT3G16130_AtRopGEF13_ KWWLPIVKVPPKGLSETLKRFLLSQRECVCQVLNSAMAINSQVLTEMEIPESYIDSLPKKGRASLGDMIYRMIT-LEMFDAEQFLLEMDLSSEHKILDLKNKFEASVVIWQRKIVQIDNK394 LOC_Os01g62990_OsRopGEF5_ KWWLPVPRVPPGGLRDKARKQLQHKRDCANQILKAALAINSNALAEMEVPESYLESLPKNGRATLGDIIYRYIT-SDHFSPECLLDCLDLSTEYQALEIANRVEASVYVWRRRIAAKPAS346 LOC_Os05g38000_OsRopGEF10_ KWWLPVPRVPPGGVGETTRRQLEHKRDCASQILKAAMAINSNALAEMDVPDSYLDSLPKNGRATLGDIIYRYIT-SDQFSPDCLLDCLDLSSEYQALEIANRVEASIYVWRRRGTSGAAS377 AT5G02010_AtRopGEF7_ KWWLPVPRVSPGGLQENSRKQLQHKRDCTNQILKAAMAINSITLADMEIPESYLESLPRKGRSCLGDLIYRYIS-SDQFSPECLLDCLDLSSEHQAIEIANRVESSIYLWHKRTNSKPAT344 AT3G55660_AtRopGEF6_ KWWLPLPRVPSDGLTEQTRKKLDHTREFTNQILKACMSINSIALAEMEVPQSYLEALPKNGRSCLGDFLYRNIT-SDNFSADHLLESIDLSSELAVVEMANRVEASMYVWRRRAHSRHLI375 AT5G05940_AtRopGEF5_ KWWLPVPRLAPNGLTEEARTELNHKRECATQILKAAMAINSLALTEMDVPKSYLETLPKNGRSCLGDVIYRYVT-SDKFSAESLLDCLDLSSEHIALDIANRVEASIYVWRRRVQTKLGV374 LOC_Os09g37270_OsRopGEF7_ KWWLPCPRVPPKGLSEDARRKLQQDRDCANQILKAAMAINSDVLAEMEIPEVYLESLPKSGKSCLGEIIYRYIT-AEQFSPECLLDCLDLSSEHHTLEVANRIEAAIHVWRLKGQKKSTP363 AT4G38430_AtRopGEF1_ KWWLPCPKVPPNGLSEEARKKLQQCRDFANQILKAALAINSGVLAEMEIPDPYLETLPKSGKECLGEIIYQYLT-ANKFSPECLLDCLDLSSEHQTLEIANRIEAAVHVWRQKNGRRHKK362 LOC_Os10g40270_OsRopGEF1_ RWWLPVPCVPDGGISGKARKELQQKRDCATQIHKAAVAINTGVLGDMEVPDSFMAILPKSGKASVGDGVYRAMLGAEKFSPEYLLDCLDMSSEHEALAMADRVEAAMYVWRRKA-GASHG389 LOC_Os04g47170_OsRopGEF9_ KWWLPVPCVTKPGLTESARRDLRQKHDCASQIHKAAMAINNGVLAEIRIPELYKQTLPKCGRASVGDLIYRHMSFPGKFSPEYLLDRLEISSEHDALEAADRVEAAMHVWRRKA-SQGHS368 LOC_Os07g29780_OsRopGEF11_ KWWLPEPCVPESGLSDRMHRELQQKRDQASQIHKMAMEINSGILSEMQVPLSYIETLPKSGKVGVGDAIYRYMSAGDQFSPDHLLNFLNLSSEHEALEIADRVEAAMYVWRRKA-SMTHV352 AT1G01700_AtRopGEF2_ KWWLPIPLVPLQGLSEKARKQLKSKRESTNQIHKAAMAINSSILGEMDIPDSYMATLPKSGKASTGDAIYRHMTSSGRFSPEKLLDRLKIVSEHEALQLADRVEASMYTWRRKA-CLNNS391 AT4G00460_AtRopGEF3_ KWWLPVPLVPSEGLSDKARKQLKNKRESTNQIHKAAMAINSSILSEMEIPDSYMTTLPKCGKSSVGDSIYRYMSGSGRFFPEQLLDCLNISSEHEAVQLADRVEASMYTWRRKS-CLSNS379 AT2G45890_AtRopGEF4_ KWWLPVPCVPAEGLSEEERKHLRHKRDCASQIHKAALAINDSTLNDMDIPDSYLTTLPKSGKASVGDVIYKQLCTAEKFYPDQLLDILKITSEHEALELADKVEASLVTWRRKTGGLTHS367 LOC_Os02g47420_OsRopGEF4_ KWWLPSPRVPDIGLSQFQRNRLVFQAKLVHQILKAAMSINEEVLLQIPIPPAVTDALPKSGRAGLGEDLYHAIT-TEYIPIEEIFLSLSLKTEHTVLETMNRLEGAVFAWNQRISEEKSK392 AT1G31650_AtRopGEF14_ RWWLPSPQVPKPGLSNSGRKKLLDKGKVVYQVFKATKAINENILLEMPVPIVIKEAIPKSGKNSLGDELYKMLA-VESATVDEIFISLNLGTEHAALETVNKLESAMFAWKERITEQGSN401 .**:* :. *: : : . . *: : ** * :: :* :*: *: *: :*: : :. :.: :* : ::.*.: *. : 104
490 500 510 520 530 540 550 560 570 580 590 600 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ TKS---SWGSAVSLEKREL---FEERAETILVLLKQKFPGLPQSSLDISKIQFNKDVGQAVLESYSRILESLAYTVMSRIEDVLYTDTLALKQTLLAEET---443
AT4G13240_AtRopGEF9_ NKS---SWGSAVSLEKREL---FEERAETILVLLKQKFPGLPQSSLDISKIQYNKDVGHAVLESYSRILESLGYTEMSRIDDVLYADSLARKQ-CTGEET---431
AT5G19560_AtRopGEF10_ GKS---QWGSTVSLEKREL---FEVRAETILVMLKQQFPGIPQSSLEVSKIKNNKDVGQAILESYSRVLESLASKIMSRIEDVLEADRLVQRQLMGEAETRS---406
LOC_Os02g17240_OsRopGEF3_ AKS---SWGSIISFEKREQ---FEERAETILHLLKLQFPGTPQSQLDISKIQYNRDVGYAILESYSRVLESLAYSVMSRIEDVLGADAAAQNLTATEAARR---425
LOC_Os05g48640_OsRopGEF2_ GKL---SWGHGVKFEKREK---FEARAENVLLLIKHRFPGIAQSALDISKIQYNRDIGLAILESYSRTLESLAFTVMSRIEDVLHADSLAQASNTRTQESMRM---475
LOC_Os01g48410_OsRopGEF6_ -KL---SWGPGVSLEKREQ---FEERAQTVLLILKHQFPGVPQSSLDISKIQYNKDVGYAILESYSRTLESLAFAVLSRIEDVLHADAIARDP--KRTKSRRR---476
LOC_Os01g55520_OsRopGEF8_ GKS---AWGSAVSQEKREQ---FEERAQTLLLIIKHRYPGIPQSTLDIAKIQENRDVGFAILESYSRVLESLAFNVMSRIEDVLNADDHAREKAKKEAPPAP---431
AT1G79860_AtRopGEF12_ SSA---PWASGVSLEKREV---FEERAETILLILKQRYPGISQSSLDISKIQFNEDVGQAVLESYSRILESLAYTVLSRIDDVLEADR---AGNKRN---443
AT1G52240_AtRopGEF11_ S---PWGSTVSIEKREQ---FEERAETILLLLKQGFPGISQSSLDISKIQFNRDVGLAILESYSRVLESLAHTVMSRIEDVLYADQLTQEPTNNAPSKNR---525
AT3G16130_AtRopGEF13_ SSS---PWSTNLSMDKRQQ---LEERAATILQLIKQEFPGISQSTLDISKIQFNRDIGLAIVESYSRILESLAHTVMSRIEDVLEADQLTQNP-ELAMCKIH---489
LOC_Os01g62990_OsRopGEF5_ VLGRATSG--RSSWGMVKDMIIDTE---KRE-LLAERAEGLLICLKQRFPGLTQTSLDMSKIQYNRDVGKSILESYSRVLESLASNIVARIDDLLNIDELNRH-AEHFP---448
LOC_Os05g38000_OsRopGEF10_ RAG----N--KSSWGIVKDMIMDTE---KRDDLLADRAEGLLMCLKQRFPGLTQTSLDTSKIQYNKDVGKSILESYSRVLESLASNIIARIDDLLYVDERSRQ-AELLPT---AG479 AT5G02010_AtRopGEF7_ NTK---TSWEMVKELMVDAD---KLE-LMADRAESLLLSLKQRFPGLPQTALDMSKIQYNKDIGKSILESYSRVLESLAFNIVARIDDLLFVDDLTRHSSDQIPT---TL444 AT3G55660_AtRopGEF6_ SLYRSTSTRWGMIVKEMMMHQTDGD---KRE-IFAERAESLLIRLKQRFPGLRQTALDTSKIQYNKDVGKSILESYSRVLESLAYSIGVRIEEVLFMDDISKDDGDGDDDSCSDKLRLLS491 AT5G05940_AtRopGEF5_ NNN-TSSTTPKLTWEMVKELMAAGD---KRG-LLVERSETLLRCLKQRFPSLTQTSLDISKIQWNKDIGKSILESYSRALESLASNIIARIDDLLYVDDLTKQSDDNN---LLS480 LOC_Os09g37270_OsRopGEF7_ QAK----SKKS-WGGKVKGLVGDTE---KSH-VLSQRADGLLQSLRLRYPGLPQTSLDMNKIQYNKDVGQSILESYSRVLESLAFNIIARIDDVIYVDDATKKS-AAADS---463
AT4G38430_AtRopGEF1_ QAK----LKLSSWGGKVKGLVNDNE---RND-FLVQRAETLLQSLRIRFPGLPQTTLDMNKIQYNKDVGQSILESYSRVMESMAFNITARIDDVLYVDDAMRRSISVTES---464
LOC_Os10g40270_OsRopGEF1_ RSR---WGAVKELVADDEE-QDKNVMLAGRAESLLLCLKHRFPGLSQTTLDTSKIQFNKDIGQAILESYSRVLESLAFNIVSWIDDVLFADKTVRKQ---482
LOC_Os04g47170_OsRopGEF9_ RSP---WSAVKELMES---DKNVMLASRAGDVLLCLKQRFPGLSQTTLDASKIQYNKDVGQAILEGYSRVLESLAYNIVTCIDDVLFADESARKI---457
LOC_Os07g29780_OsRopGEF11_ VSK---WENVTELNADG----DKNLILASRARSLLLCLKQRFPGLSQTTLDTSKIQYNKDIGQAILESYSRVLESLAYNIVSWIDDVLIADGNARKGHNIRMQ---448
AT1G01700_AtRopGEF2_ KSS---WNMVKDLMSITER-SDKNYVLAERAESLLFCLKQRYPELSQTSLDICKIHCNKDVGKAVLESYSRVLEGLAFNIVAWIDDVLYVDKTMRGEE---485
AT4G00460_AtRopGEF3_ KNS---WNMVKDLMSTTER-TDKNYVMAERAETLLFCLKQRYPELSQTSLDICKIQYNKDVGKAVLESYSRVLEGLAFNIVAWIDDVLYVDKTMRGSE---473
AT2G45890_AtRopGEF4_ KSS---WDMMKDISGDADRGNDKNHILAARARSLLFCLKQRYPELSQTSLDICKIQFNRDVGKAVLESYSRVLEGLAYNVVSWIDDVLYVDRTVRNRDD---463
LOC_Os02g47420_OsRopGEF4_ -KSPGR---HSWNFMKDSSSELD----KMSMCIERVETLMQLLKSRFPSLPPTFIEVVKIQYNVDVGHAIVEAYSRVLVGVAFSILSRVAEILLEDDLIKKPNTPLATLKFD---496
AT1G31650_AtRopGEF14_ GKSPVR---ASWSFAKDPLSEIG----RNESLLNRAEALRTQIKSKHPNLPHSFLDATKIQYDKDIGHAVLEAYSRTLANLAFRILSRMGEILKEDSLSN-PNSPAPPSCFP---505
* : :: .* : :: **: : *:* :::*.*** : .:. : ::: * 610 620 630 640 650 660 670 680 690 700 710 720 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ ---SDGG-RTTETDSESAGSSN-SGEEAEKHDPHS---KTLLDFMGWNDNSSKG-GDKPT---KSPNLTP---KKLSYLEKLENLNGFRSPKD-RH---523
AT4G13240_AtRopGEF9_ ---SDGGKIATETDSASAGSSNYSGEEIEKLESQNSSKTTLLDFIGWSDNSSKGQSEKPP---KSPRMTP---KKLSYLEKLENLNGFRSPKD-RH---517
AT5G19560_AtRopGEF10_ ----ESEAESEYEETEKVVAAETPNSRKLSDFIGWRLSSDTKKHSSMSDIEFFHKVEQEKEKP--MMKSPRALP---KKFSYLAKLENM---RSPSD-RH---493
10 20 30 40 50 60 70 80 90 100 110 120 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ ---MVAAL---ERGLSASKS---FNFKRMFDSSSTKQQQSQTIVVENGDSHIVES46 AT4G13240_AtRopGEF9_ ---MVPSL---ERGISITSS---FNLDRMFDSSPGKEQQ---QPHLAET37 AT5G19560_AtRopGEF10_ ---MFDGRNS---GHSSFSSR15 LOC_Os02g17240_OsRopGEF3_ ---MVRFL---RRHSLDKNS---AQFHRQLSHDSS---SSSSDTA33 LOC_Os05g48640_OsRopGEF2_ ---MARPL---LKIGHGLDR---FRWRRSTSSSSSSPLALSLSSSSAAALSDDDP46 LOC_Os01g48410_OsRopGEF6_ ---MVRRH---LLRGHSLDR---FLPIRSLIMSSSSSFSSS-SPSPPPPSSSSSR45 LOC_Os01g55520_OsRopGEF8_ ---MAASGGGS---LLERRSSVRRSQS---MVSEEGR28 AT1G79860_AtRopGEF12_ ---MVRAS---EQEQETYRS---RLFNFK---WRNNDN-NSATRHNKSLSVETGLDEAATG--SHDA52 AT1G52240_AtRopGEF11_ ----MLEGKAMVEDSDMPVKMQMQAMAFASQALDLFDVFDCKSIAGHIKKEFDERYGSGWQCVALLLLNYTTLIMGMPKLREEVDFNDNMNENSASRHVKSWSSDCAMRMDGSDNLDDDD116 AT3G16130_AtRopGEF13_ ---MVKAS---EKEHEKYKP---KLFDLE---NVKKKNSSSRHFKRWNSDSALRIEDPDIDDGTV53 LOC_Os01g62990_OsRopGEF5_ ---MGSG---EEERE---ARSEAAFTDSADGS----SSSSDAASADEWPV37 LOC_Os05g38000_OsRopGEF10_ ---MARGGGG---GEEAAEEERE---VSEALTADSSADEECRRGSSSSSASSGDASSE49 AT5G02010_AtRopGEF7_ ---MDGS---SENLPE---VEEKGRESSCCSSET---TRQEEEEQSPSCTE39 AT3G55660_AtRopGEF6_ MED--NSCIGFEGSKRF---GETSKRIIGL---IDSVTESTTDSSLSSSSSGVGSSSGRSSVAER57 AT5G05940_AtRopGEF5_ MENLVKSCAGIE---KKRSNL---TPSMEDILTESKERSSTSGASYESS-STTTVA49 LOC_Os09g37270_OsRopGEF7_ ---MASAS---EDDAGS---ERCCGSYSPSADVSESETSSDCSAPTTTTTTR43 AT4G38430_AtRopGEF1_ ---MGSLSS---EEDDEVS---SERCGSYSPSADISESESSSSFSCHRFDGEGA45 LOC_Os10g40270_OsRopGEF1_ ---MESSSSV---SVSDESSEANGGGCCS---SSTTPSMDAVNLSRTFSDVSSFSEEHGGSGSSVDH58 LOC_Os04g47170_OsRopGEF9_ ---MDTPS---TTCDESSEVDAR---DDYGDIDDVERRR---GRHRREASSDVSSEC45 LOC_Os07g29780_OsRopGEF11_ ---MSGGSGF---PGFHKHG---YDRDYARPLFR---VSSFSDNGGGEEQEH40 AT1G01700_AtRopGEF2_ ---MENLPNH---EENDDVG---YHQSPGPIDPNDH-SASETPVYSTMSTDSFAYH46 AT4G00460_AtRopGEF3_ ---MENLSNP---DENDD---HQSPRSIDQNDQ-SAVETPVYSTMSIDSFVYP43 AT2G45890_AtRopGEF4_ ---MESSSNS---DQNEG---TPTSSVSSPYRR-TYSDISGLSHR-FDVQSFY42 LOC_Os02g47420_OsRopGEF4_ ---MRMKTLACCRRRPQ---DFSIDMDQ-EPDRVMTYNG---LESCIINSSSYDDDSGLSATTGADGCVTTDSV64 AT1G31650_AtRopGEF14_ ---MMLMRRRLACCSRER---EISIDFDEHEQDKMITYYG---LETCIINNQSYEEESG---TSRGDGCLTDSLD63 130 140 150 160 170 180 190 200 210 220 230 240 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ NTPE---SQNSDSFVESPVE---SSLPMISPLTRPGKRSERQQADMEMMKDRFAKLLLGEDMSGGGKGVSSALALSNAITNLAA---SIF127 AT4G13240_AtRopGEF9_ TMPE---SQTQDSLGGSPVE---TSRPMTSRLIS--RRQDKQQSETEMMKDRFTKLLLGEDMSGGGKGVSSALALSNAITNLAA---SIF116 AT5G19560_AtRopGEF10_ GDGM---HTPEHELAGHAAP---STR---RGKQNRRSDMEVMKERFAKLLLGEDMSGGGTGETSALALSNAITKLAD---SMF86 LOC_Os02g17240_OsRopGEF3_ DMHE---PHGNGSGGTPPLP---NGRAAAAGAPRSRFGRDGPPSDLDVMKERFAKLLLGEDMSGTGKGVSSALALSNAITNLAA---SVF114 LOC_Os05g48640_OsRopGEF2_ GSPMDPEMPPAARRALSRSSGSRGRLSFELPPLAG---GPSDKEEAPPR---TSSAPAPARPAPAALHE-GPPSDAEMVREKFSKLLLGEDMSGTGKGVSSALALSNAITNLAA---SVF156 LOC_Os01g48410_OsRopGEF6_ GSSSGRWCGVSVAEEDEDDAAVSATTTPPLPPLQKRVLSRSHGSKAKPLGGSVDHLPPVVPPSSKNVRDSGPPSEIDLVKEKFAKLLLGEDMSGTGKGVSSALALSNAITNLAA---SVF162 LOC_Os01g55520_OsRopGEF8_ GTPA---DEDLGGGGTLKIGAVLDK---DSAAPKSRLAKDTGEHGGGGPSEMELMKEKFAKLLLGEDMSGSGKGVPSALAVSNAITNLAA---SVF115 AT1G79860_AtRopGEF12_ EPLTIIHPSQG---PPLSR---SAADEA---VLAALAASQARERQ--LLADMEQMKERFSKLLLGEDNSGGGKGVSSALALSNAITNLAA---SVF134 AT1G52240_AtRopGEF11_ NDMMMFRSQPGKCGSVDRPSLPIGGVTPNRND---KLPRVSSSDSMEALIILQAAMEQMKEKFSKLLLGEDMSGGGKGVSSALALSNAITNLAA---SAF210 AT3G16130_AtRopGEF13_ FKKTATSSIQPILPVLAMDEQPQPREATDEESQKDSGKREIISFLGYNEKFTRDQVILRFRGLQAKSCYFAYVTELEQMKDKFAKLLLGEDMSGGSKGVSSALALSNAITNLAA---SAF170 LOC_Os01g62990_OsRopGEF5_ TLAAPP--RKTAACGRVPGAEVVDSSKPHAQKRRAPS---SEMEMMKERFAKLLLGEDMSGSGKGVCTALAIANAITNLCA---TIF116 LOC_Os05g38000_OsRopGEF10_ SYCPPDEWQKVAIKTCVSSDLVVVSAEPAKEKKPPPPPSSP---RVDAAPADKHHRPSEMEMMKERFAKLLLGEDMSGGGKGVCTALAISNAITNLCA---TIF147 AT5G02010_AtRopGEF7_ DFTASP---VSSRWSVKNIDGEKKKIRSDS---RVSEVEMMKERFSKLLLGEDMSGSGNGVCTALAISNAITNLCA---TLF112 AT3G55660_AtRopGEF6_ SVSSPPTKSQILGWPLGQGSWRKSSGK---MKKKTPTKID---DFGFKRVGTETSEIELLKERMAKLLLGEDMSGSGEGVCPALAISNAITNLYA---AIL149 AT5G05940_AtRopGEF5_ SSSPPPPPSQILGWPIRKASFRKNSKESVNFDHKKLTLHD---DSGFKAKEMNSADVEMMKERFAKLLLGEDMSGSGKGVCTALAISNAITNLCA---TIF144 LOC_Os09g37270_OsRopGEF7_ RFASSSSRGVASSS---SSSLLPTPPPSSAAAFFLSAKPA---ADLSEVDMMKERFAKLLLGEDMSGSGKGVCTALAISNAITNLSA---TVF127 AT4G38430_AtRopGEF1_ SSSIPSSPRVVAGRGFYFPAPVMLPVIGGKDVVWDDKQPD---NDLSEIEMMKERFAKLLLGEDMSGGGKGVCTALAISNAITNLSA---TVF132 LOC_Os10g40270_OsRopGEF1_ SGPFEPPSAAAVSRLIGRRSPAAAASALSLSRLSMKPRADV---LDRRST-DDELELVKERFSKLLLGEDMSGGGKGVCTAVAISNAITNLYANAATVF153 LOC_Os04g47170_OsRopGEF9_ SGEPGSPYGSPYPRWP-VCSIAKAPPPPLLQKLGAARRGAG---RDRKAG-DGELQLIKERFSKLLLGEDMSGSGKGVSTAVAISNAITNLYA---TVF136 LOC_Os07g29780_OsRopGEF11_ YTP-SPPRGRSMSRTT---STAPRLSP-SVSKLSMKKLQQV---VNEKSLEDEEMELMKEKYTKLLLGEDMSGSGKGVCTAVAISNAITNLYA---TVF128 AT1G01700_AtRopGEF2_ RTCSETSGGGFSDQIDETSSFCTEASPSDWPVLTESNNSASSNFPTVFD---LKHNQIETDEHLAVQEISEPELETMKERFSKLLLGEDMSGSGKGVCTAVTISNAITNLYA---TVF158 AT4G00460_AtRopGEF3_ RTCSETTSG-FSDQIDETNSFCSEASPCNWPVLTESKSSKC---LSGLEMQSNECLVVQEISEPELETMKERFAKLLLGEDMSGSGKGVCTAVTISNAITNLYA---TVF146 AT2G45890_AtRopGEF4_ NRPSNTNAVVLSGHEEDVSEDAEEPKD---DVVNDVHGDGD---EEDSD----IDSAEDAELEMMRERFAKLLLGEDMSGSGKGVCTAVTVSNSITNLYA---TVF135 LOC_Os02g47420_OsRopGEF4_ DDEVSSCSSSKDVSSSSFSSQCHPLRKQEEHSLYELDTLSAVHLLPLKGKKPITYTLS---ASDIETMKEKFGKLLLGDDASGGARGVCAALALSNAIINLSA---TIF167 AT1G31650_AtRopGEF14_ DDAFSSCSSSKDASSS-FSSKWLPMKN-DEHSCDGLNLSGRSQHFDAKEKKKQGYGSSQHFDAKEKPGYVYCHLDVEAMKEKFSKLLLGEDVTGGCKGVQVALALSNAVTHLAT---SIF178 : :::: *****:* :* * *::::*:: :* : : 250 260 270 280 290 300 310 320 330 340 350 360 . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | . . . . | AT3G24620_AtRopGEF8_ GEQTKLQPMPQDRQARWKKEIDWLLSVTDHIVEFVPSQQTSK-DGVCTEIMVTRQRGDLLMNIPALRKLDAMLIDTLDNFRGHNEFWYVSRD---SEEGQQARNDRTND 232 AT4G13240_AtRopGEF9_ GEQTKLQPMAPDRRARWKKEIDWLLSVTDHIVEFVPSQQISK-EGVCTEIMVTRQRGDLLMNIPALRKLDAMLIDTLDNFRGHNEFWYVSRD---SEEGKQARNERTKD 221 AT5G19560_AtRopGEF10_ GEQMKLQPMYPETKENWRKEMGWLLSVIDHIVQFVPSRQMGK-NGQFTEIMVTKQRDDLLTNIPALRKLDSVLLETLDNFKDQKDFWYVPRD---MEDADHNGDWRRDE 191 LOC_Os02g17240_OsRopGEF3_ GEHRKLEPMAPDTKERWKKEVGWLLSVTDHIVEFVPTRQTAE-NGTTMEIMSTAQRRDLQMNVPALRKLDAMLIGYMDNFVDQSEFWYE---KGGDNKRDDD 212 LOC_Os05g48640_OsRopGEF2_ GEQRRLQPMAADQKARWRREIDWLLSVSDHIVEFVPSKQVSE-DGSTMEIMITQQRRDLQMNIPALRKLDAMLLEYLDNFKDKQEFWYVSKD---ASESEKGNTPRQDD 261 LOC_Os01g48410_OsRopGEF6_ GEQRRLEPMSADRRARWNKEIDWLLSVTDHIVEFVPSQQAST-DGTSMEVMGTQQRRDLLINIPALRKLDAMLLEYLDNFKDEQEFWYVKKD---ADEGEKGDAPRQGD 267 LOC_Os01g55520_OsRopGEF8_ GEQRKLEPMAPDRKGRWKKEVGWLLSVADHIVEFVAKKQVLD-NGVEMEVMGTQQRRDLQANIPALRKIDTMLLDYLDNFKDRNEFWYVKRD---S--CSDSDEQRSDE 218 AT1G79860_AtRopGEF12_ GEQRRLEPMPAERRARWRKEIDWLLSVTDYVVEFAPSQQKNK-DGTNMEIMTTRQRTDLHMNIPALKKLDAMLIDCLENFKDQSEFSYISKD---SPDL---DGKRNDE 236 AT1G52240_AtRopGEF11_ GEQRRLEPMAVDRKTRWRREIGWLISVADYIVEFAPTQQTNK-DGTSMEVMSTRQRTDLLCNIPALKKLDAMLLDCLDKFKDQDEFYYVKKD---SPDS---CETRNDE 312 AT3G16130_AtRopGEF13_ GEIRRLEAISEDKKERWRREIGWLLSVTDHIVEFSPTHQTNE-DGSSMEVMTTKQRTDLVSNIPSLKKLDEMLLDCLDKFKDQDEFYYVTPG---SPESENSNSTRNDD 275 LOC_Os01g62990_OsRopGEF5_ GQLWRLEPLPPEKKAMWRREMGWLLCVSDHIVELVPTWQSFP-DGTRLEVMTSRPRSDLYINLPALRKLDHMLIEILDSFRD-PEFWYVEQG--ICAPD---CDGSASFRAAFHRRDE 227 LOC_Os05g38000_OsRopGEF10_ GQLWRLEPLLPEKKTMWRREMDWLLCVSDHIVELVPTWQTFP-DGSKLEIMTSRPRSDLYINLPALRKLDHMLLEILESFRD-PEFWYVDQG--ICPPD---RDGSAPFMLTFHRRDE 258 AT5G02010_AtRopGEF7_ GQLWRLEPLPTEKKEMWRREMEWLLCVSDHIVEMTPTWQTFP-DGTKLEIMTCRPRSDLYVNLPALRKLDNMLLEILDSFEE-TEFWYVDQG--IMAHESA---ADGSSSFRKSFQRQED 225 AT3G55660_AtRopGEF6_ GQQWRLEPIPSEKKLMWRREIEVLLSVSDHIVELVPSFQNFP-NGNKIEVMNCRPRSDLFTCLPALRKLDNMLIEILDSFGE-TEFWYVDQG--IVAAE---SARSNSFR----EDGD 256 AT5G05940_AtRopGEF5_ GQLWRLEPLSSEKKEMWRREMEWILSVSDHIVELTPSTQTYP-DGNKFEVMTCRPRFDLFINLPALRKLDNMLLDILASFKK-TEFWYVDQG--IVASE---NDGSASFRRKIQRQEE 255 LOC_Os02g17240_OsRopGEF3_ ----LMDGDAAARKLDAMEELEKLNEAPASMTLYDFMGWHFDQDELMRKKEEGTLDEAGEAK---LKKAPSLAP---KKFSYVDSLSAG-GMRSPSA-RH---513
LOC_Os05g48640_OsRopGEF2_ --ASLSRYDTDKVVIDAKAEVERLGRMEPVSATLFDFVSPRDQDVVATKMDSKEKGCRGDAHSRKLTKVSPIAT---KRYSYLEKLENLSGTRSPIS-RH---569
LOC_Os01g48410_OsRopGEF6_ --PSLV--DIPEIIDNALEEEETVNSIDANSQVTNNSIHWQEQ---EHEDKGAR---LRKVHRMVT---KKLLHIEKVDNLGGGLKSFS-HR---554
LOC_Os01g55520_OsRopGEF8_ ----AMANDAAEHHHQQAGEVDAPCKMTGSPNGRTLLDFMDDWNGDADRPSPTAPEPAAQEDGR-LMKLPNIMTN---LKQTYMDNLFGA--HRSPPG-RH---521
AT1G79860_AtRopGEF12_ ---TPLEAE--EETLVGSMTLSDFMG---WDFDQAANAELESKKD----LPDDPLIKEKLS-VVTT---KKTSYLETLGGV---KSPTA-RH---515
AT1G52240_AtRopGEF11_ ---YSLKEN--EKLREERLSFTEDMASG---TLSDVMQWGNKNNEMKKESFFGDREKPLLSKVTG-IMTNN---KKSSYLDNLGAM---RSPTA-RYS---607
AT3G16130_AtRopGEF13_ ---IVKETESPEKEEEPNFCLLEDRPKKQKPTISLSEVMQWNIETNEPRKE----KSDKKLLTRVSSMIMSNN---KKTTYLESLGTT---RSPTAGRYS---576
LOC_Os01g62990_OsRopGEF5_ --QGDADCRIACNKAAVPPYQVP-ASGTP--FVT---AYATPSFSPAQLASP-SKKERSPLGAGRRSYSNRGFGAKKALAIDLVNPEVMGVIISGGKMIDVSTTTEL---546
LOC_Os05g38000_OsRopGEF10_ AGSGKISCMPAMSASSVPAYPVVSTSGTPPPYAT---AYATPSFSPAQLSSP-SNIGRALL-VDRRSHDGRAFDG---SMEFMGMAVSN-AVFDLPGL---568
AT5G02010_AtRopGEF7_ GNNGND--APKSIAVPVSNYTTPSYSPSKQELRS---SITVPP-SPSRFKIPHSSSVKRVLTAYVTKNEPRLKNLPLERSSRSSSSERLSLEKCMKESLNVSNLDPGI---546
AT3G55660_AtRopGEF6_ KEAASGGSGSLREKLSAPSLFSVSFSGTSTPYRTLSFSASTPSYSPMPLISP-INGGRGGERAPFLSG--RNIRER----CGFGPKKALANYLRG---579
AT5G05940_AtRopGEF5_ SPAVSS---IIAHKKVVPLPYLISASG--TPYRT-SFST-TPGFSPS-MISP-KKGER---RTPYSSKDTNKIIEKGLPSRGYGVRRVLNNYLGMESKLKICVNPSDNADTAVINQISKD588 LOC_Os09g37270_OsRopGEF7_ VSIFNRGIGVPVQKRISPSPFSIQHTPYASPFATPTFCSSTPVTGSPGRVQPPLNKDN--LPTKQEVKVEKLFSG---DIEKVWTYAGNLSARKDAGDAPERD---561
AT4G38430_AtRopGEF1_ LSLFSIN-GLNPQK---AFSVQSSPHGSPFATPALSVAS---RSP-RRAPPLYS---VKRNGTREKGIVG---ETEKAWSYAGNLSSRRVTGVTPERD---548
LOC_Os10g40270_OsRopGEF1_ ---482
LOC_Os04g47170_OsRopGEF9_ ---457
LOC_Os07g29780_OsRopGEF11_ ---KQEFSKLSPQQY---460
AT1G01700_AtRopGEF2_ ---485
AT4G00460_AtRopGEF3_ ---473
AT2G45890_AtRopGEF4_ ---463
LOC_Os02g47420_OsRopGEF4_ ----LSSDVYLAGITETPPGHIRRSLMDQISLVDG---SLDAVVRKKGVKQLRW---543
AT1G31650_AtRopGEF14_ ----SSRDPYRTPERPLLSSRVRHSLTDDMNKADGTETGLDFLFADAKASNSVNTTPSRSSRLWCLSKVPSDTSP---576
730 740 . . . . | . . . . | . . . . | . . . . | . . . AT3G24620_AtRopGEF8_ ---523
AT4G13240_AtRopGEF9_ ---517
AT5G19560_AtRopGEF10_ ---493
LOC_Os02g17240_OsRopGEF3_ ---513
LOC_Os05g48640_OsRopGEF2_ ---569
LOC_Os01g48410_OsRopGEF6_ ---554
LOC_Os01g55520_OsRopGEF8_ ---521
AT1G79860_AtRopGEF12_ ---515
AT1G52240_AtRopGEF11_ ---607
AT3G16130_AtRopGEF13_ ---576
LOC_Os01g62990_OsRopGEF5_ ---546
LOC_Os05g38000_OsRopGEF10_ ---568
AT5G02010_AtRopGEF7_ ---546
AT3G55660_AtRopGEF6_ ---579
AT5G05940_AtRopGEF5_ VEEEKKRNSTSVHQKGPPKYTVS611 LOC_Os09g37270_OsRopGEF7_ ---561
AT4G38430_AtRopGEF1_ ---548
LOC_Os10g40270_OsRopGEF1_ ---482
LOC_Os04g47170_OsRopGEF9_ ---457
LOC_Os07g29780_OsRopGEF11_ ---460
AT1G01700_AtRopGEF2_ ---485
AT4G00460_AtRopGEF3_ ---473
AT2G45890_AtRopGEF4_ ---463
LOC_Os02g47420_OsRopGEF4_ ---543
AT1G31650_AtRopGEF14_ ---576
C1
T 458 S 500 S 510
WW-motif
WW-motif
Supplementary Fig. S3
C2
C3
5
of the OsRopGEF2, OsRopGEF3, and OsRopGEF8 genes but not in OsRopGEF6. However,
K (Lysine) can also be phosphorylated.
6
Additional file 4: Figure S4. Zoom-image of subcellular localization in the tobacco epidermal cells. It shows an enlarged portion of figure 5. The first panels in left side showed control and RopGEF proteins’ GFP signal, the second panels showed RFP signal stained with the membrane marker FM4-64, the third panels showed bright image, and the last panels on the right indicate merged images. In addition to the membrane signal, the large spot seen in the RFP channel is due to Chlorophyll's auto fluorescence. a-d, Control (pGREEN-GFP); e-h, RopGEF 2; i-l, RopGEF3; m-p, RopGEF6; q-t, RopGEF8. Bars = 10 um.
GFP FM4-64 Bright field Merged
Rop GE F2 Rop GE F3 Rop GE F6 Rop GE F8 Con trol
a b
e f
i j
m n
q r
Additional file 4: Figure S4
7
Additional file 5: Figure S5. Promoter analysis of each OsRopGEF gene. Specific cis-acting element (CRE) known to affect expression in pollen were identified using PLACE database, namely three pollen-CREs and the TATA box. The number above the yellow bar shows the upstream position of the promoter base pair when taken at +1 of ATG. We analyzed up to the upstream 2,000-base pair. The p-values indicate how significantly the four genes exhibiting high expression in pollen differed from the other seven genes.
-300 -400 -600
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-2000 -1600 -1200
-400 -600
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-2000 -1600 -1200 -300
-300 -400 -600
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-2000 -1600 -1200
-300 -400 -600
-800 -200 -100
-2000 -1600 -1200
Major pollen-specific element (AGAAA) p-value = 0.30
TATA box
-300 -400 -600
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-300 -400 -600
-800 -200 -100
-2000 -1600 -1200
-300 -400 -600
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-300 -400 -600
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-300 -400 -600
-800 -200 -100
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GTGANTG10 (GTGA) p-value = 0.88 PB core (CCAC) p-value = 0.06 OsRopGEF6
OsRopGEF2
OsRopGEF8
OsRopGEF3
OsRopGEF4
OsRopGEF7
OsRopGEF10
OsRopGEF5
OsRopGEF9
OsRopGEF1
OsRopGEF11
Supplementary Fig. S4
8
Additional file 6: Figure S6. Single mutant assay using CRISPR-Cas9 system. (a-b, d-e) In- vitro pollen germination test of wild-type (a, d), ropgef2 (b, c), and ropgef3 (e-f) single mutants on solid pollen media, Bar = 50 µm; (c,f) enlarged pollen pictures, Bar = 20 µm. Each control
and mutant pollen in-vitro assays were performed in the same environment on the same day.
(g) The ratio of pollen germination. Black bar indicates wild type; grey bar indicates ropgef2;
white bar indicates ropgef3 mutant pollen. Error bars are standard deviation of three technical repeats. (h) Fertility ratio comparison between wild-type and ropgef2 single mutant plants.
Error bars are standard deviation of more than three panicles in the plants.
Additional file 6: Figure S6.
ropgef2 ropgef2
Control
a b c
d e f
ropgef3 ropgef3
Control
0 20 40 60 80 100
Control RopGEF2
Fe rt il it y rat io (% )
0 20 40 60 80 100
Germination Hydration, but no germination
No hydration
& No Germination
G e rmi na tio n rati o (% )
Control
ropgef2 ropgef3
g h
ropgef2
Control
9
Additional file 7: Figure S7. Meta-expression analysis and genome-wide identification of the seven OsRac and ten AtRop genes. (a) Heatmap expression analysis of OsRac gene. (b) Heatmap of AtRop using Genevestigator. (c) Phylogenetic tree constructs including every OsRac and AtRop.
OsRac6 OsRac5 OsRac7 OsRac3 OsRac4 OsRac1 OsRac2
a
AAF40237 AtROP3 AAF40245 AtROP5 AAC78390 AtROP1 LOC Os02g02840 OsRac6
AAF40242 AtROP6 LOC Os02g58730 OsRac5
AAC78391 AtROP2 AAF40244 AtROP4
AAF40241 AtROP7 LOC Os02g20850 OsRac7
AAD42972 AtROP8 AAF40246 AtROP9
LOC Os05g43820 OsRac2 LOC Os01g12900 OsRac1 AAF40247 AtROP10
BAB10857 AtROP11 LOC Os02g50860 OsRac3 LOC Os06g12790 OsRac4
100 70 99
50 99 72
91
99 98
77 61
66 50
80 50
0.02
c
AtRop8 AtRop3 AtRop2
AtRop7 AtRop4 AtRop5 AtRop6 AtRop1
AtRop9 AtRop10
b
5 9 13
C allus Seedling St em Shoot Leaf SAM R oot Inf lores c enc e St igm a Ov ary Seed Palea /Lem m a Spik elet Em bry o Endos perm
0% 100%
