Skip to main content

Advertisement

Springer Nature Link
Log in

CML24 is Involved in Root Mechanoresponses and Cortical Microtubule Orientation in Arabidopsis

  • Published:
Journal of Plant Growth Regulation Aims and scope Submit manuscript

Abstract

Mechanostimuli can influence plant root system architecture by causing alterations in the root tip growth direction and triggering lateral root initiation. However, how a plant root senses and translates mechanostimulation into appropriate growth and/or developmental responses remains largely unclear. The fast expression induction and transcript turnover of the Arabidopsis TCH genes by touch stimulation suggest that the TCH genes may function in mechano-related events. However, the physiological functions of the TCH genes in Arabidopsis mechanoresponses remain undetermined. Here we screened a suite of tch mutants by characterizing their root growth behaviors on hard-agar surfaces. Two calmodulin-like 24 (CML24 or TCH2) mutants, cml24-2 and cml24-4, exhibited reduced root length, biased skewing, and altered epidermal cell file rotation (CFR) phenotypes compared with wild type (Col-0). The mutant phenotypes were dependent on hard-agar surface contact and disappeared when seedlings were grown in liquid medium. Abnormal glass barrier responses of cml24 mutants further indicate touch response defects. Pharmacological tests revealed differential sensitivity of cml24 mutants to microtubule-targeted agents. Nonadditive effects of mutations in CML24 and transgenic expression of a functional microtubule label, MBD-GFP, on root skewing and CFR phenotypes suggest a potential microtubule-related role of CML24. In vivo visualization of microtubule structures with the MBD-GFP reporter revealed altered cortical microtubule orientation in the epidermal cells in cml24-4. Our observations indicate that CML24 has a role in Arabidopsis root mechanoresponses, possibly through the regulation of cortical microtubule orientation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Abe T, Hashimoto T (2005) Altered microtubule dynamics by expression of modified alpha-tubulin protein causes right-handed helical growth in transgenic Arabidopsis plants. Plant J 43:191–204

    Article  PubMed  CAS  Google Scholar 

  • Abu-Abied M, Golomb L, Belausov E, Huang SJ, Geiger B, Kam Z, Staiger CJ, Sadot E (2006) Identification of plant cytoskeleton-interacting proteins by screening for actin stress fiber association in mammalian fibroblasts. Plant J 48:367–379

    Article  PubMed  CAS  Google Scholar 

  • Braam J (1992) Regulation of expression of calmodulin and calmodulin-related genes by environmental stimuli in plants. Cell Calcium 13:457–463

    Article  PubMed  CAS  Google Scholar 

  • Braam J, Davis RW (1990) Rain-, wind-, and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. Cell 60:357–364

    Article  PubMed  CAS  Google Scholar 

  • Bruaene NV, Joss G, Oostveldt PV (2004) Reorganization and in vivo dynamics of microtubules during Arabidopsis root hair development. Plant Physiol 136:3905–3919

    Article  PubMed  Google Scholar 

  • Buer CS, Masle J, Wasteneys GO (2000) Growth conditions modulate root-wave phenotypes in Arabidopsis. Plant Cell Physiol 41:1164–1170

    Article  PubMed  CAS  Google Scholar 

  • Buer CS, Wasteneys GO, Masle J (2003) Ethylene modulates root-wave responses in Arabidopsis. Plant Physiol 132:1085–1096

    Article  PubMed  CAS  Google Scholar 

  • Chehab EW, Eich E, Braam J (2009) Thigmomorphogenesis: a complex plant response to mechano-stimulation. J Exp Bot 60:43–56

    Article  PubMed  CAS  Google Scholar 

  • Collings DA (2008) Crossed-wires: interactions and cross-talk between the microtubule and microfilament networks in plants. In: Nick P (ed) Plant microtubules 2008. Springer-Verlag, Berlin, pp 47–79

    Chapter  Google Scholar 

  • Coutand C (2010) Mechanosensing and thigmomorphogenesis, a physiological and biomechanical point of view. Plant Sci 179:168–182

    Article  CAS  Google Scholar 

  • Cyr RJ (1991) Calcium calmodulin affects microtubule stability in lysed protoplasts. J Cell Sci 100:311–317

    CAS  Google Scholar 

  • Delk NA, Johnson KA, Chowdhury NI, Braam J (2005) CML24, regulated in expression by diverse stimuli, encodes a potential Ca2+ sensor that functions in response to abscisic acid, daylength, and ion stress. Plant Physiol 139:240–253

    Article  PubMed  CAS  Google Scholar 

  • Ditengou FA, Teale WD, Kochersperger P, Flittner KA, Kneuper I, van der Graaff E, Nziengui H, Pinosa F, Li X, Nitschke R, Laux T, Palme K (2008) Mechanical induction of lateral root initiation in Arabidopsis thaliana. Proc Natl Acad Sci USA 105:18818–18823

    Article  PubMed  CAS  Google Scholar 

  • Dorlodot S, Forster B, Pages L, Price A, Tuberosa R, Draye X (2007) Root system architecture: opportunities and constraints for genetic improvement of crops. Trends Plant Sci 12:474–481

    Article  PubMed  Google Scholar 

  • Fasano JM, Massa GD, Gilroy S (2002) Ionic signaling in plant responses to gravity and touch. J Plant Growth Regul 21:71–88

    Article  PubMed  CAS  Google Scholar 

  • Fisher DD, Cyr RJ (1993) Calcium levels affect the ability to immunolocalize calmodulin to cortical microtubules. Plant Physiol 103:543–551

    PubMed  CAS  Google Scholar 

  • Fisher DD, Gilroy S, Cyr RJ (1996) Evidence for opposing effects of calmodulin on cortical microtubules. Plant Physiol 112:1079–1087

    PubMed  CAS  Google Scholar 

  • Furutani I, Watanabe Y, Prieto R, Masukawa M, Suzuki K, Naoi K, Thitamadee S, Shikanai T, Hashimoto T (2000) The SPIRAL genes are required for directional control of cell elongation in Arabidopsis thaliana. Development 127:4443–4445

    PubMed  CAS  Google Scholar 

  • Gossot O, Geitmann A (2007) Pollen tube growth: coping with mechanical obstacles involves the cytoskeleton. Planta 226:405–416

    Article  PubMed  CAS  Google Scholar 

  • Hamant O, Heisler MG, Jonsson H, Krupinski P, Uyttewaal M, Bokov P, Corson F, Sahlin P, Boudaoud A, Meyerowitz EM, Couder Y, Traas J (2008) Developmental patterning by mechanical signals in Arabidopsis. Science 322:1650–1655

    Article  PubMed  CAS  Google Scholar 

  • Hashimoto T (2002) Molecular genetic analysis of left-right handedness in plants. Philos Trans R Soc B 357:799–808

    Article  CAS  Google Scholar 

  • Heisler MG, Hamant O, Krupinski P, Uyttewaal M, Ohno C, Jonsson H, Traas J, Meyerowitz EM (2010) Alignment between PIN1 polarity and microtubule orientation in the shoot apical meristem reveals a tight coupling between morphogenesis and auxin transport. Plos Biol 8:e1000516

    Article  PubMed  Google Scholar 

  • Ishida T, Kaneko Y, Iwano M, Hashimoto T (2007a) Helical microtubule arrays in a collection of twisting tubulin mutants of Arabidopsis thaliana. Proc Natl Acad Sci USA 104:8544–8549

    Article  PubMed  CAS  Google Scholar 

  • Ishida T, Thitamadee S, Hashimoto T (2007b) Twisted growth and organization of cortical microtubules. J Plant Res 120:61–70

    Article  PubMed  CAS  Google Scholar 

  • Lee D, Polisensky DH, Braam J (2005) Genome-wide identification of touch- and darkness-regulated Arabidopsis genes: a focus on calmodulin-like and XTH genes. New Phytol 165:429–444

    Article  PubMed  CAS  Google Scholar 

  • Lloyd C, Chan J (2002) Helical microtubule arrays and spiral growth. Plant Cell 14:2319–2324

    Article  PubMed  CAS  Google Scholar 

  • Lloyd C, Chan J (2008) The parallel lives of microtubules and cellulose microfibrils. Curr Opin Plant Biol 11:641–646

    Article  PubMed  CAS  Google Scholar 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • Massa GD, Gilroy S (2003) Touch modulates gravity sensing to regulate the growth of primary roots of Arabidopsis thaliana. Plant J 33:435–445

    Article  PubMed  Google Scholar 

  • McCormack E, Braam J (2003) Calmodulins and related potential calcium sensors of Arabidopsis. New Phytol 159:585–598

    Article  CAS  Google Scholar 

  • McCormack E, Tsai YC, Braam J (2005) Handling calcium signaling: Arabidopsis CaMs and CMLs. Trends Plant Sci 10:383–389

    Article  PubMed  CAS  Google Scholar 

  • Monshausen GB, Gilroy S (2009a) The exploring root––root growth responses to local environmental conditions. Curr Opin Plant Biol 12:766–772

    Article  PubMed  CAS  Google Scholar 

  • Monshausen GB, Gilroy S (2009b) Feeling green: mechanosensing in plants. Trends Cell Biol 19:228–235

    Article  PubMed  Google Scholar 

  • Monshausen GB, Bibikova TN, Weisenseel MH, Gilroy S (2009) Ca2+ regulates reactive oxygen species production and pH during mechanosensing in Arabidopsis roots. Plant Cell 21:2341–2356

    Article  PubMed  CAS  Google Scholar 

  • Okada K, Shimura Y (1990) Reversible root-tip rotation in Arabidopsis seedlings induced by obstacle-touching stimulus. Science 250:274–276

    Article  PubMed  CAS  Google Scholar 

  • Oliva M, Dunand C (2007) Waving and skewing: how gravity and the surface of growth media affect root development in Arabidopsis. New Phytol 176:37–43

    Article  PubMed  CAS  Google Scholar 

  • Olson K, McIntosh J, Olmsted J (1995) Analysis of MAP 4 function in living cells using green fluorescent protein (GFP) chimeras. J Cell Biol 130:639–650

    Article  PubMed  CAS  Google Scholar 

  • Paredez AR, Somerville CR, Ehrhardt DW (2006) Visualization of cellulose synthase demonstrates functional association with microtubules. Science 312:1491–1495

    Article  PubMed  CAS  Google Scholar 

  • Richter GL, Monshausen GB, Krol A, Gilroy S (2009) Mechanical stimuli modulate lateral root organogenesis. Plant Physiol 151:1855–1866

    Article  PubMed  CAS  Google Scholar 

  • Rutherford R, Masson PH (1996) Arabidopsis thaliana sku mutant seedlings show exaggerated surface-dependent alteration in root growth vector. Plant Physiol 111:987–998

    Article  PubMed  CAS  Google Scholar 

  • Schwab B, Mathur J, Saedler RR, Schwarz H, Frey B, Scheidegger C, Hulskamp M (2003) Regulation of cell expansion by the distorted genes in Arabidopsis thaliana: actin controls the spatial organization of microtubules. Mol Genet Genomics 269:350–360

    Article  PubMed  CAS  Google Scholar 

  • Sedbrook JC, Kaloriti D (2008) Microtubules, MAPs and plant directional cell expansion. Trends Plant Sci 13:303–310

    Article  PubMed  CAS  Google Scholar 

  • Sugimoto K, Williamson RE, Wasteneys GO (2000) New techniques enable comparative analysis of microtubule orientation, wall texture, and growth rate in intact roots of Arabidopsis. Plant Physiol 124:1493–1506

    Article  PubMed  CAS  Google Scholar 

  • Telewski FW (2006) A unified hypothesis of mechanoperception in plants. Am J Bot 93:1466–1476

    Article  PubMed  Google Scholar 

  • Thitamadee S, Tuchihara K, Hashimoto T (2002) Microtubule basis for left-handed helical growth in Arabidopsis. Nature 417:193–196

    Article  PubMed  CAS  Google Scholar 

  • Tsai YC, Delk NA, Chowdhury NI, Braam J (2007) Arabidopsis potential calcium sensors regulate nitric oxide levels and the transition to flowering. Plant Signal Behav 2:446–454

    Article  PubMed  Google Scholar 

  • Yuen CYL, Pearlman RS, Silo-Suh L, Hilson P, Carroll KL, Masson PH (2003) WVD2 and WDL1 modulate helical organ growth and anisotropic cell expansion in Arabidopsis. Plant Physiol 131:493–506

    Article  PubMed  CAS  Google Scholar 

  • Yuen CYL, Sedbrook JC, Perrin RM, Carroll KL, Masson PH (2005) Loss-of-function mutations of root hair defective suppress root waving, skewing, and epidermal CFR in Arabidopsis. Plant Physiol 138:701–714

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This material is based upon work supported by the National Science Foundation under grant Nos. MCB 0817976 (JB) and MCB-0641288, by the National Aeronautics and Space Administration under grant No. NNX09AK80G (SG), and the National Natural Science of China under grant No. 10872223 (BW). We thank Elliot M. Meyerowitz for kindly providing the MBD-GFP seeds, and Sarah Swanson and William Deery for technical support in imaging. We are grateful to Yu-Chang Tsai and Won-Gyu Choi for helpful discussions and to Liz Eich for isolation and characterization of the cml12-2 mutant.

Author information

Authors and Affiliations

  1. Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, Bioengineering College, Chongqing University, Chongqing, 400030, China

    Yichuan Wang & Bochu Wang

  2. Department of Biochemistry and Cell Biology, Rice University, Houston, TX, 77005, USA

    Yichuan Wang, E. Wassim Chehab & Janet Braam

  3. Department of Botany, University of Wisconsin at Madison, Madison, WI, 53706, USA

    Simon Gilroy

Authors
  1. Yichuan Wang
    View author publications

    Search author on:PubMed Google Scholar

  2. Bochu Wang
    View author publications

    Search author on:PubMed Google Scholar

  3. Simon Gilroy
    View author publications

    Search author on:PubMed Google Scholar

  4. E. Wassim Chehab
    View author publications

    Search author on:PubMed Google Scholar

  5. Janet Braam
    View author publications

    Search author on:PubMed Google Scholar

Corresponding authors

Correspondence to Bochu Wang or Janet Braam.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (AVI 451 kb) Time-lapse movie of a wild-type root as in Figure 2a, showing a typical step-like growth response upon encountering a glass barrier. Time interval between fames is 10 minutes. Scale bars= 1mm

Supplementary material 2 (AVI 453 kb) Time-lapse movie of a cml24-2 root as in Figure 2f, showing an aberrant growth response upon encountering a glass barrier. Time interval between fames is 10 minutes. Scale bars= 1mm

Supplementary material 3 (AVI 426 kb) Time-lapse movie of a cml24-4 root as in Figure 2g, showing an aberrant growth response upon encountering a glass barrier. Time interval between fames is 10 minutes. Scale bars= 1mm

344_2011_9209_MOESM4_ESM.tif

Supplementary material 4 (TIFF 4203 kb) Root waving of wild type, cam2-1, and cml12-2 grown on hard-agar surfaces. (a) Wild type Col-0 seedlings grown on hard-agar surfaces tilted 15o backward from vertical. No detectable biased root skewing or general root growth defect was found in (b) cam2-1 or (c) cml12-2 seedlings compared with (a) wild type in the same condition. Images were taken from above the agar surface. Scale bars: a-c = 10 mm

344_2011_9209_MOESM5_ESM.tif

Supplementary material 5 (TIFF 5086 kb) Root cell file features of wild type, cml24-2, and cml24-4 grown on hard-agar surfaces supplemented with 1µM taxol. Representative root cell file features of (a) wild type, (b) cml24-2, and (c) cml24-4 when grown on 1µM taxol supplemented hard-agar surfaces tilted 15o backward from vertical. No swollen cells were found in (a) wild type or (c) cml24-4, whereas many swollen cells were detected in (b) cml24-2. Scale bars: a-c = 0.5mm

344_2011_9209_MOESM6_ESM.tif

Supplementary material 6 (TIFF 2859 kb) Root waving and skewing of wild type, cml24-2, and wild-type (Col-0) YC3.6, YC3.6/cml24-2 grown on hard-agar surfaces. YC3.6 is a transgenic line producing a Ca2+ reporter (Monshausen and others 2009.) (a) Wild type Col-0, (b) cml24-2, (c) YC3.6, and (d) YC3.6/cml24-2 seedlings grown on hard-agar surfaces tilted 15o backward from vertical. No detectable rescue of the general growth defect was found in cml24-2 seedlings (b) expressing YC3.6 (d). Images were taken from above the agar surface. Scale bars: a-d = 10mm

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, Y., Wang, B., Gilroy, S. et al. CML24 is Involved in Root Mechanoresponses and Cortical Microtubule Orientation in Arabidopsis . J Plant Growth Regul 30, 467–479 (2011). https://doi.org/10.1007/s00344-011-9209-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00344-011-9209-9

Keywords