A noncanonical auxin-sensing mechanism is required for organ morphogenesis in Arabidopsis

Tissue patterning in multicellular organisms is the output of precise spatio–temporal regulation of gene expression coupled with changes in hormone dynamics. In plants, the hormone auxin regulates growth and development at every stage of a plant's life cycle. Auxin signaling occurs through binding of the auxin molecule to a TIR1/AFB F-box ubiquitin ligase, allowing interaction with Aux/IAA transcriptional repressor proteins. These are subsequently ubiquitinated and degraded via the 26S proteasome, leading to derepression of auxin response factors (ARFs). How auxin is able to elicit such a diverse range of developmental responses through a single signaling module has not yet been resolved. Here we present an alternative auxin-sensing mechanism in which the ARF ARF3/ETTIN controls gene expression through interactions with process-specific transcription factors. This noncanonical hormone-sensing mechanism exhibits strong preference for the naturally occurring auxin indole 3-acetic acid (IAA) and is important for coordinating growth and patterning in diverse developmental contexts such as gynoecium morphogenesis, lateral root emergence, ovule development, and primary branch formation. Disrupting this IAA-sensing ability induces morphological aberrations with consequences for plant fitness. Therefore, our findings introduce a novel transcription factor-based mechanism of hormone perception in plants. Note that there is a CORRIGENDUM to this article: http://eprints.whiterose.ac.uk/132306/ http://genesdev.cshlp.org/content/31/17/1821.ful

DELLA-Induced Early Transcriptional Changes during Etiolated Development in Arabidopsis thaliana

The hormones gibberellins (GAs) control a wide variety of processes in plants, including stress and developmental responses. This task largely relies on the activity of the DELLA proteins, nuclear-localized transcriptional regulators that do not seem to have DNA binding capacity. The identification of early target genes of DELLA action is key not only to understand how GAs regulate physiological responses, but also to get clues about the molecular mechanisms by which DELLAs regulate gene expression. Here, we have investigated the global, early transcriptional response triggered by the Arabidopsis DELLA protein GAI during skotomorphogenesis, a developmental program tightly regulated by GAs. Our results show that the induction of GAI activity has an almost immediate effect on gene expression. Although this transcriptional regulation is largely mediated by the PIFs and HY5 transcription factors based on target meta-analysis, additional evidence points to other transcription factors that would be directly involved in DELLA regulation of gene expression. First, we have identified cis elements recognized by Dofs and type-B ARRs among the sequences enriched in the promoters of GAI targets; and second, an enrichment in additional cis elements appeared when this analysis was extended to a dataset of early targets of the DELLA protein RGA: CArG boxes, bound by MADS-box proteins, and the E-box CACATG that links the activity of DELLAs to circadian transcriptional regulation. Finally, Gene Ontology analysis highlights the impact of DELLA regulation upon the homeostasis of the GA, auxin, and ethylene pathways, as well as upon pre-existing transcriptional networks

The bHLH transcription factor SPATULA enables cytokinin signaling, and both activate auxin biosynthesis and transport genes at the medial domain of the gynoecium

[EN] Fruits and seeds are the major food source on earth. Both derive from the gynoecium and, therefore, it is crucial to understand the mechanisms that guide the development of this organ of angiosperm species. In Arabidopsis, the gynoecium is composed of two congenitally fused carpels, where two domains: medial and lateral, can be distinguished. The medial domain includes the carpel margin meristem (CMM) that is key for the production of the internal tissues involved in fertilization, such as septum, ovules, and transmitting tract. Interestingly, the medial domain shows a high cytokinin signaling output, in contrast to the lateral domain, where it is hardly detected. While it is known that cytokinin provides meristematic properties, understanding on the mechanisms that underlie the cytokinin signaling pattern in the young gynoecium is lacking. Moreover, in other tissues, the cytokinin pathway is often connected to the auxin pathway, but we also lack knowledge about these connections in the young gynoecium. Our results reveal that cytokinin signaling, that can provide meristematic properties required for CMM activity and growth, is enabled by the transcription factor SPATULA (SPT) in the medial domain. Meanwhile, cytokinin signaling is confined to the medial domain by the cytokinin response repressor ARABIDOPSIS HISTIDINE PHOSPHOTRANSFERASE 6 (AHP6), and perhaps by ARR16 (a type-A ARR) as well, both present in the lateral domains (presumptive valves) of the developing gynoecia. Moreover, SPT and cytokinin, probably together, promote the expression of the auxin biosynthetic gene TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (TAA1) and the gene encoding the auxin efflux transporter PIN-FORMED 3 (PIN3), likely creating auxin drainage important for gynoecium growth. This study provides novel insights in the spatiotemporal determination of the cytokinin signaling pattern and its connection to the auxin pathway in the young gynoecium.IRO, VMZM, HHU and PLS were supported by the Mexican National Council of Science and Technology (CONACyT) with a PhD fellowship (210085, 210100, 243380 and 219883, respectively). Work in the SDF laboratory was financed by the CONACyT grants CB-2012-177739, FC-2015-2/1061, and INFR-2015-253504, and NMM by the CONACyT grant CB-2011-165986. SDF, CF and LC acknowledge the support of the European Union FP7-PEOPLE-2009-IRSES project EVOCODE (grant no. 247587) and H2020-MSCARISE-2015 project ExpoSEED (grant no. 691109). SDF also acknowledges the Marine Biological Laboratory (MBL) in Woods Hole for a scholarship for the Gene Regulatory Networks for Development Course 2015 (GERN2015). IE acknowledges the International European Fellowship-METMADS project and the Universita degli Studi di Milano (RTD-A; 2016). Research in the laboratory of MFY was funded by NSF (grant IOS-1121055), NIH (grant 1R01GM112976-01A1) and the Paul D. Saltman Endowed Chair in Science Education (MFY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Reyes Olalde, J.; Zuñiga, V.; Serwatowska, J.; Chávez Montes, R.; Lozano-Sotomayor, P.; Herrera-Ubaldo, H.; Gonzalez Aguilera, K.... (2017). The bHLH transcription factor SPATULA enables cytokinin signaling, and both activate auxin biosynthesis and transport genes at the medial domain of the gynoecium. PLoS Genetics. 13(4):1-31. https://doi.org/10.1371/journal.pgen.1006726S131134Reyes-Olalde, J. I., Zuñiga-Mayo, V. M., Chávez Montes, R. A., Marsch-Martínez, N., & de Folter, S. (2013). Inside the gynoecium: at the carpel margin. Trends in Plant Science, 18(11), 644-655. doi:10.1016/j.tplants.2013.08.002Alvarez-Buylla, E. R., Benítez, M., Corvera-Poiré, A., Chaos Cador, Á., de Folter, S., Gamboa de Buen, A., … Sánchez-Corrales, Y. E. (2010). Flower Development. The Arabidopsis Book, 8, e0127. doi:10.1199/tab.0127Bowman, J. L., Baum, S. F., Eshed, Y., Putterill, J., & Alvarez, J. (1999). 4 Molecular Genetics of Gynoecium Development in Arabidopsis. Current Topics in Developmental Biology Volume 45, 155-205. doi:10.1016/s0070-2153(08)60316-6Chávez Montes, R. A., Herrera-Ubaldo, H., Serwatowska, J., & de Folter, S. (2015). Towards a comprehensive and dynamic gynoecium gene regulatory network. Current Plant Biology, 3-4, 3-12. doi:10.1016/j.cpb.2015.08.002Marsch-Martínez, N., & de Folter, S. (2016). Hormonal control of the development of the gynoecium. Current Opinion in Plant Biology, 29, 104-114. doi:10.1016/j.pbi.2015.12.006Marsch-Martínez, N., Ramos-Cruz, D., Irepan Reyes-Olalde, J., Lozano-Sotomayor, P., Zúñiga-Mayo, V. M., & de Folter, S. (2012). The role of cytokinin during Arabidopsis gynoecia and fruit morphogenesis and patterning. The Plant Journal, 72(2), 222-234. doi:10.1111/j.1365-313x.2012.05062.xZhao, Z., Andersen, S. U., Ljung, K., Dolezal, K., Miotk, A., Schultheiss, S. J., & Lohmann, J. U. (2010). Hormonal control of the shoot stem-cell niche. Nature, 465(7301), 1089-1092. doi:10.1038/nature09126Ashikari, M. (2005). Cytokinin Oxidase Regulates Rice Grain Production. Science, 309(5735), 741-745. doi:10.1126/science.1113373Bartrina, I., Otto, E., Strnad, M., Werner, T., & Schmülling, T. (2011). Cytokinin Regulates the Activity of Reproductive Meristems, Flower Organ Size, Ovule Formation, and Thus Seed Yield in Arabidopsis thaliana. The Plant Cell, 23(1), 69-80. doi:10.1105/tpc.110.079079Hwang, I., Sheen, J., & Müller, B. (2012). Cytokinin Signaling Networks. Annual Review of Plant Biology, 63(1), 353-380. doi:10.1146/annurev-arplant-042811-105503Schaller, G. E., Bishopp, A., & Kieber, J. J. (2015). The Yin-Yang of Hormones: Cytokinin and Auxin Interactions in Plant Development. The Plant Cell, 27(1), 44-63. doi:10.1105/tpc.114.133595Kieber, J. J., & Schaller, G. E. (2010). The Perception of Cytokinin: A Story 50 Years in the Making: Figure 1. Plant Physiology, 154(2), 487-492. doi:10.1104/pp.110.161596Long, J. A., Moan, E. I., Medford, J. I., & Barton, M. K. (1996). A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature, 379(6560), 66-69. doi:10.1038/379066a0Jasinski, S., Piazza, P., Craft, J., Hay, A., Woolley, L., Rieu, I., … Tsiantis, M. (2005). KNOX Action in Arabidopsis Is Mediated by Coordinate Regulation of Cytokinin and Gibberellin Activities. Current Biology, 15(17), 1560-1565. doi:10.1016/j.cub.2005.07.023Yanai, O., Shani, E., Dolezal, K., Tarkowski, P., Sablowski, R., Sandberg, G., … Ori, N. (2005). Arabidopsis KNOXI Proteins Activate Cytokinin Biosynthesis. Current Biology, 15(17), 1566-1571. doi:10.1016/j.cub.2005.07.060Scofield, S., Dewitte, W., Nieuwland, J., & Murray, J. A. H. (2013). The Arabidopsis homeobox gene SHOOT MERISTEMLESS has cellular and meristem-organisational roles with differential requirements for cytokinin and CYCD3 activity. The Plant Journal, 75(1), 53-66. doi:10.1111/tpj.12198Gordon, S. P., Chickarmane, V. S., Ohno, C., & Meyerowitz, E. M. (2009). Multiple feedback loops through cytokinin signaling control stem cell number within the Arabidopsis shoot meristem. Proceedings of the National Academy of Sciences, 106(38), 16529-16534. doi:10.1073/pnas.0908122106Chickarmane, V. S., Gordon, S. P., Tarr, P. T., Heisler, M. G., & Meyerowitz, E. M. (2012). Cytokinin signaling as a positional cue for patterning the apical-basal axis of the growing Arabidopsis shoot meristem. Proceedings of the National Academy of Sciences, 109(10), 4002-4007. doi:10.1073/pnas.1200636109Leibfried, A., To, J. P. C., Busch, W., Stehling, S., Kehle, A., Demar, M., … Lohmann, J. U. (2005). WUSCHEL controls meristem function by direct regulation of cytokinin-inducible response regulators. Nature, 438(7071), 1172-1175. doi:10.1038/nature04270Werner, T., Motyka, V., Laucou, V., Smets, R., Van Onckelen, H., & Schmülling, T. (2003). Cytokinin-Deficient Transgenic Arabidopsis Plants Show Multiple Developmental Alterations Indicating Opposite Functions of Cytokinins in the Regulation of Shoot and Root Meristem Activity. The Plant Cell, 15(11), 2532-2550. doi:10.1105/tpc.014928Larsson, E., Franks, R. G., & Sundberg, E. (2013). Auxin and the Arabidopsis thaliana gynoecium. Journal of Experimental Botany, 64(9), 2619-2627. doi:10.1093/jxb/ert099Weijers, D., & Wagner, D. (2016). Transcriptional Responses to the Auxin Hormone. Annual Review of Plant Biology, 67(1), 539-574. doi:10.1146/annurev-arplant-043015-112122Robert, H. S., Crhak Khaitova, L., Mroue, S., & Benková, E. (2015). The importance of localized auxin production for morphogenesis of reproductive organs and embryos inArabidopsis. Journal of Experimental Botany, 66(16), 5029-5042. doi:10.1093/jxb/erv256Kuusk, S., Sohlberg, J. J., Magnus Eklund, D., & Sundberg, E. (2006). Functionally redundantSHIfamily genes regulate Arabidopsis gynoecium development in a dose-dependent manner. The Plant Journal, 47(1), 99-111. doi:10.1111/j.1365-313x.2006.02774.xSohlberg, J. J., Myrenås, M., Kuusk, S., Lagercrantz, U., Kowalczyk, M., Sandberg, G., & Sundberg, E. (2006). STY1regulates auxin homeostasis and affects apical-basal patterning of the Arabidopsis gynoecium. The Plant Journal, 47(1), 112-123. doi:10.1111/j.1365-313x.2006.02775.xStåldal, V., Sohlberg, J. J., Eklund, D. M., Ljung, K., & Sundberg, E. (2008). Auxin can act independently ofCRC,LUG,SEU,SPTandSTY1in style development but not apical-basal patterning of theArabidopsisgynoecium. New Phytologist, 180(4), 798-808. doi:10.1111/j.1469-8137.2008.02625.xVan Gelderen, K., van Rongen, M., Liu, A., Otten, A., & Offringa, R. (2016). An INDEHISCENT-Controlled Auxin Response Specifies the Separation Layer in Early Arabidopsis Fruit. Molecular Plant, 9(6), 857-869. doi:10.1016/j.molp.2016.03.005José Ripoll, J., Bailey, L. J., Mai, Q.-A., Wu, S. L., Hon, C. T., Chapman, E. J., … Yanofsky, M. F. (2015). microRNA regulation of fruit growth. Nature Plants, 1(4). doi:10.1038/nplants.2015.36Larsson, E., Roberts, C. J., Claes, A. R., Franks, R. G., & Sundberg, E. (2014). Polar Auxin Transport Is Essential for Medial versus Lateral Tissue Specification and Vascular-Mediated Valve Outgrowth in Arabidopsis Gynoecia. Plant Physiology, 166(4), 1998-2012. doi:10.1104/pp.114.245951Nole-Wilson, S., Azhakanandam, S., & Franks, R. G. (2010). Polar auxin transport together with AINTEGUMENTA and REVOLUTA coordinate early Arabidopsis gynoecium development. Developmental Biology, 346(2), 181-195. doi:10.1016/j.ydbio.2010.07.016De Folter, S. (2016). Auxin Is Required for Valve Margin Patterning in Arabidopsis After All. Molecular Plant, 9(6), 768-770. doi:10.1016/j.molp.2016.05.005Moubayidin, L., & Østergaard, L. (2014). Dynamic Control of Auxin Distribution Imposes a Bilateral-to-Radial Symmetry Switch during Gynoecium Development. Current Biology, 24(22), 2743-2748. doi:10.1016/j.cub.2014.09.080Girin, T., Paicu, T., Stephenson, P., Fuentes, S., Körner, E., O’Brien, M., … Østergaard, L. (2011). INDEHISCENT and SPATULA Interact to Specify Carpel and Valve Margin Tissue and Thus Promote Seed Dispersal in Arabidopsis. The Plant Cell, 23(10), 3641-3653. doi:10.1105/tpc.111.090944Ioio, R. D., Nakamura, K., Moubayidin, L., Perilli, S., Taniguchi, M., Morita, M. T., … Sabatini, S. (2008). A Genetic Framework for the Control of Cell Division and Differentiation in the Root Meristem. Science, 322(5906), 1380-1384. doi:10.1126/science.1164147Bishopp, A., Help, H., El-Showk, S., Weijers, D., Scheres, B., Friml, J., … Helariutta, Y. (2011). A Mutually Inhibitory Interaction between Auxin and Cytokinin Specifies Vascular Pattern in Roots. Current Biology, 21(11), 917-926. doi:10.1016/j.cub.2011.04.017De Rybel, B., Adibi, M., Breda, A. S., Wendrich, J. R., Smit, M. E., Novák, O., … Weijers, D. (2014). Integration of growth and patterning during vascular tissue formation in Arabidopsis. Science, 345(6197), 1255215. doi:10.1126/science.1255215Pernisova, M., Klima, P., Horak, J., Valkova, M., Malbeck, J., Soucek, P., … Hejatko, J. (2009). Cytokinins modulate auxin-induced organogenesis in plants via regulation of the auxin efflux. Proceedings of the National Academy of Sciences, 106(9), 3609-3614. doi:10.1073/pnas.0811539106Cheng, Z. J., Wang, L., Sun, W., Zhang, Y., Zhou, C., Su, Y. H., … Zhang, X. S. (2012). Pattern of Auxin and Cytokinin Responses for Shoot Meristem Induction Results from the Regulation of Cytokinin Biosynthesis by AUXIN RESPONSE FACTOR3. Plant Physiology, 161(1), 240-251. doi:10.1104/pp.112.203166Alvarez, J., & Smyth, D. R. (2002). CRABS CLAWandSPATULAGenes Regulate Growth and Pattern Formation during Gynoecium Development inArabidopsis thaliana. International Journal of Plant Sciences, 163(1), 17-41. doi:10.1086/324178Groszmann, M., Bylstra, Y., Lampugnani, E. R., & Smyth, D. R. (2010). Regulation of tissue-specific expression of SPATULA, a bHLH gene involved in carpel development, seedling germination, and lateral organ growth in Arabidopsis. Journal of Experimental Botany, 61(5), 1495-1508. doi:10.1093/jxb/erq015Smyth, D. R., Bowman, J. L., & Meyerowitz, E. M. (1990). Early flower development in Arabidopsis. The Plant Cell, 2(8), 755-767. doi:10.1105/tpc.2.8.755Müller, B., & Sheen, J. (2008). Cytokinin and auxin interaction in root stem-cell specification during early embryogenesis. Nature, 453(7198), 1094-1097. doi:10.1038/nature06943Argyros, R. D., Mathews, D. E., Chiang, Y.-H., Palmer, C. M., Thibault, D. M., Etheridge, N., … Schaller, G. E. (2008). Type B Response Regulators of Arabidopsis Play Key Roles in Cytokinin Signaling and Plant Development. The Plant Cell, 20(8), 2102-2116. doi:10.1105/tpc.108.059584Mason, M. G., Mathews, D. E., Argyros, D. A., Maxwell, B. B., Kieber, J. J., Alonso, J. M., … Schaller, G. E. (2005). Multiple Type-B Response Regulators Mediate Cytokinin Signal Transduction in Arabidopsis. The Plant Cell, 17(11), 3007-3018. doi:10.1105/tpc.105.035451Ishida, K., Yamashino, T., Yokoyama, A., & Mizuno, T. (2008). Three Type-B Response Regulators, ARR1, ARR10 and ARR12, Play Essential but Redundant Roles in Cytokinin Signal Transduction Throughout the Life Cycle of Arabidopsis thaliana. Plant and Cell Physiology, 49(1), 47-57. doi:10.1093/pcp/pcm165Yokoyama, A., Yamashino, T., Amano, Y.-I., Tajima, Y., Imamura, A., Sakakibara, H., & Mizuno, T. (2006). Type-B ARR Transcription Factors, ARR10 and ARR12, are Implicated in Cytokinin-Mediated Regulation of Protoxylem Differentiation in Roots of Arabidopsis thaliana. Plant and Cell Physiology, 48(1), 84-96. doi:10.1093/pcp/pcl040Schuster, C., Gaillochet, C., & Lohmann, J. U. (2015). Arabidopsis HECATE genes function in phytohormone control during gynoecium development. Development, 142(19), 3343-3350. doi:10.1242/dev.120444Toledo-Ortiz, G., Huq, E., & Quail, P. H. (2003). The Arabidopsis Basic/Helix-Loop-Helix Transcription Factor Family. The Plant Cell, 15(8), 1749-1770. doi:10.1105/tpc.013839Reymond, M. C., Brunoud, G., Chauvet, A., Martínez-Garcia, J. F., Martin-Magniette, M.-L., Monéger, F., & Scutt, C. P. (2012). A Light-Regulated Genetic Module Was Recruited to Carpel Development in Arabidopsis following a Structural Change to SPATULA. The Plant Cell, 24(7), 2812-2825. doi:10.1105/tpc.112.097915Ballester, P., Navarrete-Gómez, M., Carbonero, P., Oñate-Sánchez, L., & Ferrándiz, C. (2015). Leaf expansion in Arabidopsis is controlled by a TCP-NGA regulatory module likely conserved in distantly related species. Physiologia Plantarum, 155(1), 21-32. doi:10.1111/ppl.12327Hellens, R., Allan, A., Friel, E., Bolitho, K., Grafton, K., Templeton, M., … Laing, W. (2005). Plant Methods, 1(1), 13. doi:10.1186/1746-4811-1-13Makkena, S., & Lamb, R. S. (2013). The bHLH transcription factor SPATULA regulates root growth by controlling the size of the root meristem. BMC Plant Biology, 13(1), 1. doi:10.1186/1471-2229-13-1Stepanova, A. N., Robertson-Hoyt, J., Yun, J., Benavente, L. M., Xie, D.-Y., Doležal, K., … Alonso, J. M. (2008). TAA1-Mediated Auxin Biosynthesis Is Essential for Hormone Crosstalk and Plant Development. Cell, 133(1), 177-191. doi:10.1016/j.cell.2008.01.047Bhargava, A., Clabaugh, I., To, J. P., Maxwell, B. B., Chiang, Y.-H., Schaller, G. E., … Kieber, J. J. (2013). Identification of Cytokinin-Responsive Genes Using Microarray Meta-Analysis and RNA-Seq in Arabidopsis. Plant Physiology, 162(1), 272-294. doi:10.1104/pp.113.217026Sakai, H., Aoyama, T., & Oka, A. (2000). Arabidopsis ARR1 and ARR2 response regulators operate as transcriptional activators. The Plant Journal, 24(6), 703-711. doi:10.1046/j.1365-313x.2000.00909.xSakai, H. (2001). ARR1, a Transcription Factor for Genes Immediately Responsive to Cytokinins. Science, 294(5546), 1519-1521. doi:10.1126/science.1065201Moubayidin, L., Di Mambro, R., Sozzani, R., Pacifici, E., Salvi, E., Terpstra, I., … Sabatini, S. (2013). Spatial Coordination between Stem Cell Activity and Cell Differentiation in the Root Meristem. Developmental Cell, 26(4), 405-415. doi:10.1016/j.devcel.2013.06.025Benková, E., Michniewicz, M., Sauer, M., Teichmann, T., Seifertová, D., Jürgens, G., & Friml, J. (2003). Local, Efflux-Dependent Auxin Gradients as a Common Module for Plant Organ Formation. Cell, 115(5), 591-602. doi:10.1016/s0092-8674(03)00924-3Okada, K., Ueda, J., Komaki, M. K., Bell, C. J., & Shimura, Y. (1991). Requirement of the Auxin Polar Transport System in Early Stages of Arabidopsis Floral Bud Formation. The Plant Cell, 677-684. doi:10.1105/tpc.3.7.677Blilou, I., Xu, J., Wildwater, M., Willemsen, V., Paponov, I., Friml, J., … Scheres, B. (2005). The PIN auxin efflux facilitator network controls growth and patterning in Arabidopsis roots. Nature, 433(7021), 39-44. doi:10.1038/nature03184Mahonen, A. P. (2006). Cytokinin Signaling and Its Inhibitor AHP6 Regulate Cell Fate During Vascular Development. Science, 311(5757), 94-98. doi:10.1126/science.1118875Besnard, F., Refahi, Y., Morin, V., Marteaux, B., Brunoud, G., Chambrier, P., … Vernoux, T. (2013). Cytokinin signalling inhibitory fields provide robustness to phyllotaxis. Nature, 505(7483), 417-421. doi:10.1038/nature12791Longabaugh, W. J. R., Davidson, E. H., & Bolouri, H. (2005). Computational representation of developmental genetic regulatory networks. Developmental Biology, 283(1), 1-16. doi:10.1016/j.ydbio.2005.04.023Faure, E., Peter, I. S., & Davidson, E. H. (2013). A New Software Package for Predictive Gene Regulatory Network Modeling and Redesign. Journal of Computational Biology, 20(6), 419-423. doi:10.1089/cmb.2012.0297Mangan, S., & Alon, U. (2003). Structure and function of the feed-forward loop network motif. Proceedings of the National Academy of Sciences, 100(21), 11980-11985. doi:10.1073/pnas.2133841100Chen, Q., Liu, Y., Maere, S., Lee, E., Van Isterdael, G., Xie, Z., … Vanneste, S. (2015). A coherent transcriptional feed-forward motif model for mediating auxin-sensitive PIN3 expression during lateral root development. Nature Communications, 6(1). doi:10.1038/ncomms9821Qiu, K., Li, Z., Yang, Z., Chen, J., Wu, S., Zhu, X., … Zhou, X. (2015). EIN3 and ORE1 Accelerate Degreening during Ethylene-Mediated Leaf Senescence by Directly Activating Chlorophyll Catabolic Genes in Arabidopsis. PLOS Genetics, 11(7), e1005399. doi:10.1371/journal.pgen.1005399Seaton, D. D., Smith, R. W., Song, Y. H., MacGregor, D. R., Stewart, K., Steel, G., … Halliday, K. J. (2015). Linked circadian outputs control elongation growth and flowering in response to photoperiod and temperature. Molecular Systems Biology, 11(1), 776. doi:10.15252/msb.20145766Roeder, A. H. K., & Yanofsky, M. F. (2006). Fruit Development in Arabidopsis. The Arabidopsis Book, 4, e0075. doi:10.1199/tab.0075Marsch-Martínez, N., Reyes-Olalde, J. I., Ramos-Cruz, D., Lozano-Sotomayor, P., Zúñiga-Mayo, V. M., & de Folter, S. (2012). Hormones talking. Plant Signaling & Behavior, 7(12), 1698-1701. doi:10.4161/psb.22422Balanza, V., Navarrete, M., Trigueros, M., & Ferrandiz, C. (2006). Patterning the female side of Arabidopsis: the importance of hormones. Journal of Experimental Botany, 57(13), 3457-3469. doi:10.1093/jxb/erl188Kamiuchi, Y., Yamamoto, K., Furutani, M., Tasaka, M., & Aida, M. (2014). The CUC1 and CUC2 genes promote carpel margin meristem formation during Arabidopsis gynoecium development. Frontiers in Plant Science, 5. doi:10.3389/fpls.2014.00165Scofield, S., Dewitte, W., & Murray, J. A. H. (2007). The KNOX gene SHOOT MERISTEMLESS is required for the development of reproductive meristematic tissues in Arabidopsis. The Plant Journal, 50(5), 767-781. doi:10.1111/j.1365-313x.2007.03095.xLi, K., Yu, R., Fan, L.-M., Wei, N., Chen, H., & Deng, X. W. (2016). DELLA-mediated PIF degradation contributes to coordination of light and gibberellin signalling in Arabidopsis. Nature Communications, 7(1). doi:10.1038/ncomms11868Oh, E., Zhu, J.-Y., & Wang, Z.-Y. (2012). Interaction between BZR1 and PIF4 integrates brassinosteroid and environmental responses. Nature Cell Biology, 14(8), 802-809. doi:10.1038/ncb2545Sharma, N., Xin, R., Kim, D.-H., Sung, S., Lange, T., & Huq, E. (2016). NO FLOWERING IN SHORT DAY (NFL) is a bHLH transcription factor that promotes flowering specifically under short-day conditions inArabidopsis. Development, 143(4), 682-690. doi:10.1242/dev.128595Varaud, E., Brioudes, F., Szécsi, J., Leroux, J., Brown, S., Perrot-Rechenmann, C., & Bendahmane, M. (2011). AUXIN RESPONSE FACTOR8 Regulates Arabidopsis Petal Growth by Interacting with the bHLH Transcription Factor BIGPETALp. The Plant Cell, 23(3), 973-983. doi:10.1105/tpc.110.081653Savaldi-Goldstein, S., & Chory, J. (2008). Growth coordination and the shoot epidermis. Current Opinion in Plant Biology, 11(1), 42-48. doi:10.1016/j.pbi.2007.10.009Schuster, C., Gaillochet, C., Medzihradszky, A., Busch, W., Daum, G., Krebs, M., … Lohmann, J. U. (2014). A Regulatory Framework for Shoot Stem Cell Co

Spatio-temporal sequence of cross-regulatory events in root meristem growth

A central question in developmental biology is how multicellular organisms coordinate cell division and differentiation to determine organ size. In Arabidopsis roots, this balance is controlled by cytokinin-induced expression of SHORT HYPOCOTYL 2 (SHY2) in the so-called transition zone of the meristem, where SHY2 negatively regulates auxin response factors (ARFs) by protein–protein interaction. The resulting down-regulation of PIN-FORMED (PIN) auxin efflux carriers is considered the key event in promoting differentiation of meristematic cells. Here we show that this regulation involves additional, intermediary factors and is spatio-temporally constrained. We found that the described cytokinin–auxin crosstalk antagonizes BREVIS RADIX (BRX) activity in the developing protophloem. BRX is an auxin-responsive target of the prototypical ARF MONOPTEROS (MP), a key promoter of vascular development, and transiently enhances PIN3 expression to promote meristem growth in young roots. At later stages, cytokinin induction of SHY2 in the vascular transition zone restricts BRX expression to down-regulate PIN3 and thus limit meristem growth. Interestingly, proper SHY2 expression requires BRX, which could reflect feedback on the auxin responsiveness of SHY2 because BRX protein can directly interact with MP, likely acting as a cofactor. Thus, cross-regulatory antagonism between BRX and SHY2 could determine ARF activity in the protophloem. Our data suggest a model in which the regulatory interactions favor BRX expression in the early proximal meristem and SHY2 prevails because of supplementary cytokinin induction in the later distal meristem. The complex equilibrium of this regulatory module might represent a universal switch in the transition toward differentiation in various developmental contexts

Corrigendum: A noncanonical auxin-sensing mechanism is required for organ morphogenesis in Arabidopsis (vol 30, pg 2286, 2017)

We have realized that there is an error in Figure 4 of the above-mentioned article. By mistake, the images in Figure 4, A and D, originate from the same sample. This image is from the untreated control experiment (Fig. 4A) but was mistakenly also used in Figure 4D (2,4-D treatment). We have built a revised Figure 4 in which we corrected this error. Figure 4. View larger version: In this page In a new window Download as PowerPoint Slide Figure 4. ETT and IND protein interact in an IAA-sensitive manner. (A–D) BiFC with pYFPN-ETT and pYFPC-IND in the absence (A) and presence of IAA (B), NAA (C), and 2,4-D (D). All hormonal treatments were with 1 mM in lanolin. (E) Fluorescence quantification (CTCF) of split YFP signal between ETT and IND without treatment and with IAA, NAA, and 2,4-D. (*) P < 0.01; (***) P < 0.0001. Error bars show the standard deviation. (F) Y2H assays with increasing concentration of IAA, NAA, and 2,4D. Bars: A–D, 50 μm. Moreover, we have added a raw data file as an Additional Supplemental Material link associated with the original article with the raw images of three independent experiments that were used for the quantification in Figure 4E (Figure_4_raw_data.pdf). For each condition, we have included a picture taken with a low magnification (a 10× objective). In addition, we have included four more images at higher magnification (a 20× objective) and have repeated the intensity quantification using the nuclei that are labeled with a number. The values are summarized below the images, and a new chart has been built with these data, including a new ANOVA analysis. These data show the same result as in the original Figure 4