Cell production and differentiation for the acquisition of specific functions are key features of living systems. The dynamic network of cellular microtubules provides the necessary platform to accommodate processes associated with the transition of cells through the individual phases of cytogenesis. Here, we show that the plant hormone cytokinin fine-tunes the activity of the microtubular cytoskeleton during cell differentiation and counteracts microtubular rearrangements driven by the hormone auxin. The endogenous upward gradient of cytokinin activity along the longitudinal growth axis in Arabidopsis thaliana roots correlates with robust rearrangements of the microtubule cytoskeleton in epidermal cells progressing from the proliferative to the differentiation stage. Controlled increases in cytokinin activity result in premature re-organization of the microtubule network from transversal to an oblique disposition in cells prior to their differentiation, whereas attenuated hormone perception delays cytoskeleton conversion into a configuration typical for differentiated cells. Intriguingly, cytokinin can interfere with microtubules also in animal cells, such as leukocytes, suggesting that a cytokinin-sensitive control pathway for the microtubular cytoskeleton may be at least partially conserved between plant and animal cells.

Bioresources Unit Center for Health and Bioresources AIT Austrian Institute of Technology GmbH Tulln Austria

Department of Plant Biotechnology and Bioinformatics Ghent University and Center for Plant Systems Biology VIB Gent Belgium

Institute of Experimental Botany The Czech Academy of Sciences Praha Czech Republic

Institute of Science and Technology Austria Klosterneuburg Austria

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Adamowski M, Li L, Friml J (2019) Reorientation of cortical microtubule arrays in the hypocotyl of PubMed PMC

Akhmanova A, Steinmetz MO (2008) Tracking the ends: A dynamic protein network controls the fate of microtubule tips. Nat Rev Mol Cell Biol 9: 309–322 PubMed

Akhmanova A, Steinmetz MO (2015) Control of microtubule organization and dynamics: two ends in the limelight. Nat Rev Mol Cell Biol 16: 711–726 PubMed

Antoniadi I, Pla L (2015) Cell‐type‐specific cytokinin distribution within the PubMed PMC

Applegate KT, Besson S, Matov A, Bagonis MH, Jaqaman K, Danuser G (2011) PlusTipTracker: quantitative image analysis software for the measurement of microtubule dynamics. J Struct Biol 176: 168–184 PubMed PMC

Argueso CT, Ferreira FJ, Kieber JJ (2009) Environmental perception avenues: the interaction of cytokinin and environmental response pathways. Plant, Cell Environ 32: 1147–1160 PubMed

Argyros RD, Mathews DE, Chiang YH, Palmer CM, Thibault DM, Etheridge N, Argyros DA, Mason MG, Kieber JJ, Schallera GE (2008) Type B response regulators of PubMed PMC

Attwood PV (2013) Histidine kinases from bacteria to humans. Biochem Soc Trans 41: 1023–1028 PubMed

Baluška F, Mancuso S, Volkmann D, Barlow PW (2010) Root apex transition zone: a signalling‐response nexus in the root. Trends Plant Sci 15: 402–408 PubMed

Bao Y, Kost B, Chua NH (2001) Reduced expression of α‐tubulin genes in PubMed

Baskin TI, Wilson JE, Cork A, Williamson RE (1994) Morphology and microtubule organization in PubMed

Baskin TI, Beemster GTS, Judy‐March JE, Marga F (2004) Disorganization of cortical microtubules stimulates tangential expansion and reduces the uniformity of cellulose microfibril alignment among cells in the root of PubMed PMC

Bichet A, Desnos T, Turner S, Grandjean O, Höfte H (2001) BOTERO1 is required for normal orientation of cortical microtubules and anisotropic cell expansion in PubMed

Bielach A, Podlešáková K, Marhavý P, Duclercq J, Cuesta C, Müller B, Grunewald W, Tarkowski P, Benkováa E (2012) Spatiotemporal regulation of lateral root organogenesis in PubMed PMC

Bifulco M, Malfitano AM, Proto MC, Santoro A, Caruso MG, Laezza C (2008) Biological and pharmacological roles of N6‐isopentenyladenosine: an emerging anticancer drug. Anticancer Agents Med Chem 8: 200–204 PubMed

Billou I, Xu J, Wildwater M, Willemsen V, Paponov I, Frimi J, Heldstra R, Aida M, Palme K, Scheres B (2005) The PIN auxin efflux facilitator network controls growth and patterning in PubMed

Bishopp A, Help H, El‐Showk S, Weijers D, Scheres B, Friml J, Benková E, Mähönen AP, Helariutta Y (2011) A mutually inhibitory interaction between auxin and cytokinin specifies vascular pattern in roots. Curr Biol 21: 917–926 PubMed

Boudaoud A, Burian A, Borowska‐Wykrȩt D, Uyttewaal M, Wrzalik R, Kwiatkowska D, Hamant O (2014) FibrilTool, an ImageJ plug‐in to quantify fibrillar structures in raw microscopy images. Nat Protoc 9: 457–463 PubMed

Brandizzi F, Wasteneys GO (2013) Cytoskeleton‐dependent endomembrane organization in plant cells: an emerging role for microtubules. Plant J 75: 339–349 PubMed

Burk DH, Liu B, Zhong R, Morrison WH, Ye ZH (2001) A katanin‐like protein regulates normal cell wall biosynthesis and cell elongation. Plant Cell 13: 807–827 PubMed PMC

Buschmann H, Sambade A, Pesquet E, Calder G, Lloyd CW (2010) Microtubule dynamics in plant cells. Methods Cell Biol 97: 373–400 PubMed

Casati S, Ottria R, Baldoli E, Lopez E, Maier JAM, Ciuffreda P (2011) Effects of cytokinins, cytokinin ribosides and their analogs on the viability of normal and neoplastic human cells. Anticancer Res 31: 3401–3406 PubMed

Chandler JW, Werr W (2015) Cytokinin‐auxin crosstalk in cell type specification. Trends Plant Sci 20: 291–300 PubMed

Chen X, Grandont L, Li H, Hauschild R, Paque S, Abuzeineh A, Rakusová H, Benková E, Perrot‐Rechenmann C, Friml JJ PubMed PMC

Davidse L (1986) Benzimidazole fungicides: mechanism of action and biological impact. Ann Rev Phytopathol 24: 43–65

De Rybel B, Adibi M, Breda AS, Wendrich JR, Smit ME, Novák O, Yamaguchi N, Yoshida S, Van Isterdael G, Palovaara J PubMed

Dello Ioio R, Nakamura K, Moubayidin L, Perilli S, Taniguchi M, Morita MT, Aoyama T, Costantino P, Sabatini S (2008) A genetic framework for the control of cell division and differentiation in the root meristem. Science (80‐.) 322: 1380–1384 PubMed

Di Mambro R, De Ruvo M, Pacifici E, Salvi E, Sozzani R, Benfey PN, Busch W, Novak O, Ljung K, Di Paola L PubMed PMC

Dixit R, Chang E, Cyr RJ (2006) Establishment of polarity during organization of the acentrosomal plant cortical microtubule array. Mol Biol Cell 17: 1298–1305 PubMed PMC

Elliott A, Shaw SL (2018) Update: plant cortical microtubule arrays. Plant Physiol 176: 94–105 PubMed PMC

Etienne‐Manneville S (2010) From signaling pathways to microtubule dynamics: the key players. Curr Opin Cell Biol 22: 104–111 PubMed

Geldner N, Dénervaud‐Tendon V, Hyman DL, Mayer U, Stierhof YD, Chory J (2009) Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. Plant J 59: 169–178 PubMed PMC

Griffaut B, Bos R, Maurizis JC, Madelmont JC, Ledoigt G (2004) Cytotoxic effects of kinetin riboside on mouse, human and plant tumour cells. Int J Biol Macromol 34: 271–275 PubMed

Gupta S, Rashotte AM (2012) Down‐stream components of cytokinin signaling and the role of cytokinin throughout the plant. Plant Cell Rep 31: 801–812 PubMed

Gutierrez R, Lindeboom JJ, Paredez AR, Emons AMC, Ehrhardt DW (2009) PubMed

Hamada T (2014) Microtubule organization and microtubule‐associated proteins in plant cells. Int Rev Cell Mol Biol 312: 1–52 PubMed

Hare PD, Cress WA, Van Staden J (1997) The involvement of cytokinins in plant responses to environmental stress. Plant Growth Regul 23: 79–103

Hashimoto T (2015) Microtubules in plants. Arabidopsis Book 13: e0179 PubMed PMC

Hayashi K, Hasegawa J, Matsunaga S (2013) The boundary of the meristematic and elongation zones in roots: endoreduplication precedes rapid cell expansion. Sci Rep 3: 1–8 PubMed PMC

Higuchi M, Pischke MS, Mähönen AP, Miyawaki K, Hashimoto Y, Seki M, Kobayashi M, Shinozaki K, Kato T, Tabata S PubMed PMC

Hönig M, Plíhalová L, Spíchal L, Grúz J, Kadlecová A, Voller J, Svobodová AR, Vostálová J, Ulrichová J, Doležal K PubMed

Horio T, Murata T (2014) The role of dynamic instability in microtubule organization. Front Plant Sci 5: 511 PubMed PMC

Hugdahl JD, Morejohn LC (1993) Rapid and reversible high‐affinity binding of the dinitroaniline herbicide oryzalin to tubulin from PubMed PMC

Inoue T, Higuchi M, Hashimoto Y, Seki M, Kobayashi M, Kato T, Tabata S, Shinozaki K, Kakimoto T (2001) Identification of CRE1 as a cytokinin receptor from PubMed

Ishida T, Hashimoto T (2007) An PubMed

Ishida T, Kaneko Y, Iwano M, Hashimoto T (2007) Helical microtubule arrays in a collection of twisting tubulin mutants of PubMed PMC

Ishii Y, Sakai S, Honma Y (2003) Cytokinin‐induced differentiation of human myeloid leukemia HL‐60 cells is associated with the formation of nucleotides, but not with incorporation into DNA or RNA. Biochim Biophys Acta Mol Cell Res 1643: 11–24 PubMed

Ivanov VB, Dubrovsky JG (2013) Longitudinal zonation pattern in plant roots: conflicts and solutions. Trends Plant Sci 18: 237–243 PubMed

Jabłońska‐Trypuć A, Matejczyk M, Czerpak R (2016) N6‐benzyladenine and kinetin influence antioxidative stress parameters in human skin fibroblasts. Mol Cell Biochem 413: 97–107 PubMed PMC

Jakobs MA, Dimitracopoulos A, Franze K (2019) KymoButler, a deep learning software for automated kymograph analysis. Elife 8: 1–19 PubMed PMC

Kadlecová A, Maková B, Artal‐Sanz M, Strnad M, Voller J (2019) The plant hormone kinetin in disease therapy and healthy aging. Ageing Res Rev 55: 100958 PubMed

de Keijzer J, Mulder BM, Janson ME (2014) Microtubule networks for plant cell division. Syst Synth Biol 8: 187–194 PubMed PMC

Kopf A, Renkawitz J, Hauschild R, Girkontaite I, Tedford K, Merrin J, Thorn‐Seshold O, Trauner D, Häcker H, Fischer K‐D PubMed DOI PMC

Laplaze L, Benkova E, Casimiro I, Maes L, Vanneste S, Swarup R, Weijers D, Calvo V, Parizot B, Herrera‐Rodriguez MB PubMed PMC

Le J, Vandenbussche F, Van Der Straeten D, Verbelen JP (2004) Position and cell type‐dependent microtubule reorientation characterizes the early response of the

Le J, Vandenbussche F, De Cnodder T, Van Der Straeten D, Verbelen JP (2005) Cell elongation and microtubule behavior in the

Lee YC, Yang YC, Huang CL, Kuo TY, Lin JH, Yang DM, Huang NK (2012) When cytokinin, a plant hormone, meets the adenosine A2A receptor: a novel neuroprotectant and lead for treating neurodegenerative disorders? PLoS ONE 7: e38865 PubMed PMC

Li S, Sun T, Ren H (2015) The functions of the cytoskeleton and associated proteins during mitosis and cytokinesis in plant cells. Front Plant Sci 6: 282 PubMed PMC

Löfke C, Scheuring D, Dünser K, Schöller M, Luschnig C, Kleine‐Vehn J (2015) Tricho‐ and atrichoblast cell files show distinct PIN2 auxin efflux carrier exploitations and are jointly required for defined auxin‐dependent root organ growth. J Exp Bot 66: 5103–5112 PubMed PMC

Lucas J, Shaw SL (2008) Cortical microtubule arrays in the PubMed

Marc J, Granger CL, Brincat J, Fisher DD, Kao TH, McCubbin AG, Cyr RJ (1998) A GFP‐MAP4 reporter gene for visualizing cortical microtubule rearrangements in living epidermal cells. Plant Cell 10: 1927–1939 PubMed PMC

Marhavý P, Bielach A, Abas L, Abuzeineh A, Duclercq J, Tanaka H, Pařezová M, Petrášek J, Friml J, Kleine‐Vehn J PubMed

Martín‐Cófreces NB, Sánchez‐Madrid F (2018) Sailing to and docking at the immune synapse: role of tubulin dynamics and molecular motors. Front Immunol 9: 1–15 PubMed PMC

Morejohn LC, Fosket DE (1991) The biochemistry of compounds with anti‐microtubule activity in plant cells. Pharmacol Ther 51: 217–230 PubMed

Morejohn LC, Bureau TE, Mole‐Bajer J, Bajer AS, Fosket DE (1987) Oryzalin, a dinitroaniline herbicide, binds to plant tubulin and inhibits microtubule polymerization PubMed

Mostowy S, Shenoy AR (2015) The cytoskeleton in cell‐autonomous immunity: structural determinants of host defence. Nat Rev Immunol 15: 559–573 PubMed PMC

Moubayidin L, Di Mambro R, Sabatini S (2009) Cytokinin‐auxin crosstalk. Trends Plant Sci 14: 557–562 PubMed

Müller B, Sheen J (2008) Cytokinin and auxin interaction in root stem‐cell specification during early embryogenesis. Nature 453: 1094–1097 PubMed PMC

Nick P, Schäfer E, Furuya M (1992) Auxin redistribution during first positive phototropism in corn coleoptiles: microtubule Reorientation and the Cholodny‐Went theory. Plant Physiol 99: 1302–1308 PubMed PMC

Nishimura C, Ohashi Y, Sato S, Kato T, Tabata S, Ueguchi C (2004) Histidine kinase homologs that act as cytokinin receptors possess overlapping functions in the regulation of shoot and root growth in PubMed PMC

O'Brien JA, Benková E (2013) Cytokinin cross‐talking during biotic and abiotic stress responses. Front Plant Sci 4: 1–11 PubMed PMC

Oda Y (2015) Cortical microtubule rearrangements and cell wall patterning. Front Plant Sci 6: 1–7 PubMed PMC

Othman EM, Naseem M, Awad E, Dandekar T, Stopper H (2016) The plant hormone cytokinin confers protection against oxidative stress in mammalian cells. PLoS ONE 11: 1–14 PubMed PMC

Panteris E, Diannelidis B‐E, Adamakis I‐DS (2018) Cortical microtubule orientation in PubMed PMC

Pasternak T, Tietz O, Rapp K, Begheldo M, Nitschke R, Ruperti B, Palme K (2015) Protocol: an improved and universal procedure for whole‐mount immunolocalization in plants. Plant Methods 11: 1–10 PubMed PMC

Pavelescu I, Vilarrasa‐Blasi J, Planas‐Riverola A, González‐García M, Caño‐Delgado AI, Ibañes M (2018) A Sizer model for cell differentiation in PubMed PMC

Petersson SV, Johansson AI, Kowalczyk M, Makoveychuk A, Wang JY, Moritz T, Grebe M, Benfey PN, Sandberg G, Ljung K (2009) An auxin gradient and maximum in the PubMed PMC

Petricka JJ, Winter CM, Benfey PN (2012) Control of PubMed PMC

Rasmussen CG, Wright AJ, Müller S (2013) The role of the cytoskeleton and associated proteins in determination of the plant cell division plane. Plant J 75: 258–269 PubMed

Reinhardt D, Mandel T, Kuhlemeier C (2000) Auxin regulates the initiation and radial position of plant lateral organs. Plant Cell 12: 507–518 PubMed PMC

Renkawitz J, Kopf A, Stopp J, de Vries I, Driscoll MK, Merrin J, Hauschild R, Welf ES, Danuser G, Fiolka R PubMed PMC

Ruzicka K, Simásková M, Duclercq J, Petrásek J, Zazímalová E, Simon S, Friml J, Van Montagu MCE, Benková E (2009) Cytokinin regulates root meristem activity via modulation of the polar auxin transport. Proc Natl Acad Sci USA 106: 4284–4289 PubMed PMC

Sakai H, Aoyama T, Oka A (2000) Arabidopsis ARR1 and ARR2 response regulators operate as transcriptional activators. Plant J 24: 703–711 PubMed

Sakai H, Honma T, Takashi A, Sato S, Kato T, Tabata S, Oka A (2001) ARR1, a transcription factor for genes immediately responsive to cytokinins. Science (80‐.) 294: 1519–1521 PubMed

Salazar‐Henao JE, Vélez‐Bermúdez IC, Schmidt W (2016) The regulation and plasticity of root hair patterning and morphogenesis. Development 143: 1848–1858 PubMed

Samakovli D, Komis G (2017) Katanin effects on dynamics of cortical microtubules and mitotic arrays in PubMed PMC

Schaller GE, Street IH, Kieber JJ (2014) Cytokinin and the cell cycle. Curr Opin Plant Biol 21: 7–15 PubMed

Schaller GE, Bishopp A, Kieber JJ (2015) The Yin‐Yang of hormones: cytokinin and auxin interactions in plant development. Plant Cell 27: 44–63 PubMed PMC

Shibata M, Sugimoto K (2019) A gene regulatory network for root hair development. J Plant Res 132: 301–309 PubMed PMC

Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured PubMed

Slovak R, Ogura T, Satbhai SB, Ristova D, Busch W (2016) Genetic control of root growth: from genes to networks. Ann Bot 117: 9–24 PubMed PMC

Smertenko A, Assaad F, Baluška F, Bezanilla M, Buschmann H, Drakakaki G, Hauser MT, Janson M, Mineyuki Y, Moore I PubMed

Smet W, Sevilem I, de Luis Balaguer MA, Wybouw B, Mor E, Miyashima S, Blob B, Roszak P, Jacobs TB, Boekschoten M PubMed PMC

Sozzani R, Iyer‐Pascuzzi A (2014) Postembryonic control of root meristem growth and development. Curr Opin Plant Biol 17: 7–12 PubMed

Sugimoto K, Williamson RE, Wasteneys GO (2000) New techniques enable comparative analysis of microtubule orientation, wall texture, and growth rate in intact roots of PubMed PMC

Takahashi H, Kawahara A, Inoue Y (2003) Ethylene promotes the induction by auxin of the cortical microtubule randomization required for low‐pH‐induced root hair initiation in lettuce ( PubMed

Takatsuka H, Umeda M (2014) Hormonal control of cell division and elongation along differentiation trajectories in roots. J Exp Bot 65: 2633–2643 PubMed

Takesue K, Shibaoka H (1998) The cyclic reorientation of cortical microtubules in epidermal cells of azuki bean epicotyls: the role of actin filaments in the progression of the cycle. Planta 205: 539–546 PubMed

Tiedemann RE, Mao X, Shi CX, Yuan XZ, Palmer SE, Sebag M, Marler R, Chesi M, Fonseca R, Bergsagel PL PubMed PMC

Tinevez JY, Perry N, Schindelin J, Hoopes GM, Reynolds GD, Laplantine E, Bednarek SY, Shorte SL, Eliceiri KW (2017) TrackMate: an open and extensible platform for single‐particle tracking. Methods 115: 80–90 PubMed

To JPC, Kieber JJ (2008) Cytokinin signaling: two‐components and more. Trends Plant Sci 13: 85–92 PubMed

True JH, Shaw SL (2020) Exogenous auxin induces transverse microtubule arrays through transport inhibitor response1/auxin signaling F‐BOX receptors. Plant Physiol 182: 892–907 PubMed PMC

Verbelen JP, De Cnodder T, Le J, Vissenberg K, Baluška F (2006) The root apex of PubMed PMC

Vicente‐Manzanares M, Sánchez‐Madrid F (2004) Role of the cytoskeleton during leukocyte responses. Nat Rev Immunol 4: 110–122 PubMed

Vilches Barro A, Stöckle D, Thellmann M, Ruiz‐Duarte P, Bald L, Louveaux M, von Born P, Denninger P, Goh T, Fukaki H PubMed

Vineyard L, Elliott A, Dhingra S, Lucas JR, Shaw SL (2013) Progressive transverse microtubule array organization in hormone‐induced PubMed PMC

von Wangenheim D, Hauschild R, Fendrych M, Barone V, Benková E, Friml J (2017) Live tracking of moving samples in confocal microscopy for vertically grown roots. Elife 6: e26792 PubMed PMC

Wasteneys GO (2002) Microtubule organization in the green kingdom: chaos or self‐order? J Cell Sci 115: 1345–1354 PubMed

Werner T, Motyka V, Laucou V, Smets R, Van Onckelen H, Schmülling T (2003) Cytokinin‐deficient transgenic PubMed PMC

Wong JH, Hashimoto T (2017) Novel PubMed PMC

Zürcher E, Tavor‐Deslex D, Lituiev D, Enkerli K, Tarr PT, Müller B (2013) A robust and sensitive synthetic sensor to monitor the transcriptional output of the cytokinin signaling network in planta. Plant Physiol 161: 1066–1075 PubMed PMC

GR24, A Synthetic Strigolactone Analog, and Light Affect the Organization of Cortical Microtubules in Arabidopsis Hypocotyl Cells