Arabidopsis Embryogenesis & &> & # «^ f í Comparison of embryo development in Angiosperms Caryophyllad Chenopodiad Solanad Asterad iHĚfc, ;M^ p© ® 1 I 3 © 1 |® ® I ®1 © @ ' Jllk & # E5«J itbiKll feirsii Modified after Johrietal. 1992 How can such a protected system be investigated experimentally? Mutant screen at seedling level Patterning mutant types APICAL (gurke) CENTRAL (fackel) 64SAL (monopterous) TERMINAL © (gnom) Mutations in the BODENLOS (bdi) and MONOPTEROS (mp) genes lead to very similar deletions of basal pattern elements mp seedling bdl seedling 3 bodenlos (bdi) root meristem defect octant globular heart Model of BDL/MP interaction Eh MP Inhibited Auxin MP Active BDL Degradation (Proteasome) Root Meristem Initiation Genetic Interference with Auxin Response and Transport Disrupts Embryo Patterning monopteros bodenlos gnom DR5::GFP in Embryo Mutants Auxin signaling Auxin transport BFA treatment monopteros gnom DR5::GFP- in vitro Culturing Long time treatments Control NAA 2,4D NPA or BFA Auxin Transport Chemiosmotic hypothesis Cell wall-Plasma membrane Efflux carrier------ PIN-family Influx ___ carrier AUX1 IAAH pH=7.0 IAA"+H pH=5.5 Arabidopsis PIN Protein Family Phylogenetic tree Homology of PIN proteins AtPINI AtPiN2 ^ AtPIN6 AtPIN4 AtPIN7 -**1 AtPIN3 AIPIN8 ^^AtPIN5 ÍBn\ třmí 121 ; t FIVi 14Í tPIHJ 3IJ lfL.MlMVPLÍVIlMIHíCÍVH(lí : : T" P a QÍIÍŮ 1 NXÍVIl, ľ*VÍ Ľ, LS l"H ľ 11 Ü H Y. řřJU "■HJlv L ptT v>M i L »t c k v RWBiL: r:; pn Qtiů i nnrv*: r»vB ľ,íS r« r i ít »Hr rju >■.■ v ľ L s-VAM i J. Ri-Ľ Ä v . W1É ÍL l r:; PĎQťSC IÉ ArVX::. fcví Ll,i FM r r (T H L přJU »HTťLÍVAML ll-LiKVKKWlllFTk&OC A I HA I.VM - ÍT LtL h T I i: FT ä H ITfrS | ah .-!■ LT >M: -v lli C i-V K.H -Jtí Ft F,-. QCSŮI HJinf j; víäVÍ vLSFi F1 tu H*i*T--: I y :, I rXMI Lfci C SVJtHHa L F:; FEhQCtC IH A TV A : tKVt L LS f M F11 ü njiíIX: a?V»L4U:; Kt t- r:. Ja« :i :.K±. f-í i k QCAfl IJUifViUiTü i WLÍtW: i If Ji W9r.'.\ 'IŕLb:AKr&~TA«tLHV^ĽVVLOciTWTT^LLTLTIvFCAKLLtM| víla KAKXftOv * ■■":- L v f a ::'V^QjAI v*: TLUifv: irfcuJl IřLt-AKr&UTT^TLWOi.vvLOCIIHTTLLLrLTi: p.-Aii : [j"(.l.:Aicfíir-T55íl-KVfl]vv^5cl""rLi.Lj-LrivR5JL«:i:!'is: MPiL.-ft:TarTnA|ir.r.giWLa-.MHriTUrLn:.NAftRAL?.':^ _ . 5IHlf5PJl5í|i5 . >■ a:; ■:h: .-.*v::x:■ nr.:;»:.-;.-.:;htt .p|rc... .Ifk.. h| kŠfhi F. r|ttľ5 ,i ICKFSPWTCCGCCTJ HKTO. . . stěcabcvI E It 9 TTT STAU K B-V n| |e|. STCTCVSTEP A ABC C VEK ■ RHU F 5 PRHGT TiTST ■ ŮVDWIHI......................^^.....................IgBBRITWVgZKa —13 SJTE IlFC HŮD LQ 0 Ti? FT R PQ Lbe FR^IBSTfA. S Sľ"----- . pjarp.............■.. .flpvp hDviTivgiiRom Hfjs^ eha s 5 fr; ■ BIMJ.EE AH I........................................ZDEp(ZETĎBlv|l v RTF! HAFV.........VSfJJAQ KE nm sc acvtv SKÜ . FK DVNTNŮG"TLFT 2H 9 HRH Dftl PEE. . . NŮC S L-Q E K D S|a . SH D*) SV§THrt . tPIWfl 210 . STUHETC-SPJA PE E . KLIĚDHTC.JCEHI 7¥N :•■ .JcphhcetI. tPIJ15 192 tPXHS 21a 4 2 3 EEFSFQlNftDfiEMKUL. . A?2 (GH NI £jtKT|...............ŮAKV1 tlJ KQ........HVpH£QbSNttl. . . . CÄSFÍrtCKKCSaYEBGCŕCŕRXQO i 1* í Q F£ FACaE E E Alp. FK ĽAEhIlH KLA FN ÍJfEALŮS KTC LCŮAEASŮEKW LUSCEŮEREí|k. . . aTa|lM KMC SN sH* LiAACCfiCŮSHifC. . Tn| - ■ K OIIJ^l 11 'JPVKKBkM ![ L ľ nvtíxK L ľ RH v: : L : KV ■■.,■ k i: kľi IL1MUWIRLIAHPI VK^Lnnii-jii VH:! KLI Lv v V: URL ■: RH PUT X S I YMTJtL I LIHVHR.RLIAM FHT X 3 ILIraWRRLI : M FHT f a kAKf3L21F HAHFSLŮ1F M UHF Ä J. B L ľ si ÍJs$ V*I:QAALPqcIVFFVHXtTMWHFÖItttaVIfCMLIUK»: TT JUflL--k i vflAA L pqi IV prvmíi Tirv h po i LITaII j rcrn-.n pv Iv :.xt vttflt - ■ AIVUAALÍQÍlVPTVrHAETXVHPAinLSTČVITtiKLIALPirL-.ITCLLSL »IVillíLigClírrVrUlITXVIIfľlLITCTlTCKLIHLUrĽ.-IICLlSL :VAI:'QfLH.lřflMT!;r[rWttV::i.HACivLtfAVir«hLv. " " a:: ľ L ha hl VflA» L pqí IV FTVr» m.| tH :. h FO iKf :.V I t ::k : v. C-CLLÄVAIVŮA* L FOÍ "í FTVr*ÄÉ tltVh FAI tiTÍV t Fí Ml. I At FIŤ! v«i:.j*»Ltqs-:jnvr>Mry:.iiľ.l:tti;¥lTaKLl>Lť:l PIN1 in E GUS mRNA PIN7 in Embryogenesis GUS mRNA Protein PIN1 T*^*. Phenotypes of pin Multiple Mutants 4x7 1x3x 1x3x4x x4x. 1x3x4x7 1x3x4x7 Auxin and Apical pole specification BĎL PIN1 PIN7 Two-Cell Embryogenesis Root pole specification GNOM PIN7 Globular Auxin in Flower and DR5rev::GFP DR5 in Ovule Formation Ovule with Integuments primordia Ovule defects in pin1 PIN1 in Ovule Formation Gynoecium with ovule primordia Ovule primordium Ovule with Integuments primordia Auxin \r\ Lateral Root Development ÍŮ(£ii ÍŮMji ivi al ty Correlates with Root Initiation 11 15h 1 B Oh i 15h j ■jí w V i ■■■.'' i ^** B ! I lil unpublished DR5 in Lateral Root Formation DR5rev::GUS IAA pin1,3,4 Lateral Root Development Organogenic process involving re-entry into cell cycle and coordinated cell divisions and differentiation. Initiation (in pericycle) and development phases can be distinguished. Both phases require both long and short distance signaling probably by auxin and cytokinin. The lateral root meristem development is mediated by auxin gradient. Common module for organ formation Aerial organogenesis Underground organogenesis Cotyledons, leaves, flowers, floral organs, ovules, integuments Lateral roots Root Meristem Light microscopy Confocal microscopy X^Iejíti and phloem Fe«eycte I Corte* km iai I H Gorton frnduderns Oun-r l B*r"ipyif NHaftdRH epidermis Lalcri root "P Columella růů1 cap Fin. 4> A simpk HIChIl-J represcn tinji [wo difícn:nl nc£u|jiory *ipn:iE^ wiíbin (hü fOOI JttcritfL-in. "ETic qutr*Lbni ixnEn; inhibits Jilľ-Ľn:nLĽt[ion