Abstract
γ-Tubulin complex protein 4 (GCP4, encoded by AT3G53760) participates in microtubule (MT) nucleation in Arabidopsis thaliana, affecting the MT nucleation angles in cortical MTs, and the formation of the spindle and phragmoplasts during mitosis. Here, we report that GCP4 plays a critical role in gametophyte development. The results indicate that the gcp4 mutant caused by T-DNA insertion may express an aberrant gene product interfering with normal GCP4 expression, ultimately leading to the formation of desiccated ovules and aborted seeds. An analysis of transmission efficiency (TE) indicated that female gametophytes were more impaired in development than male gametophytes, and so observation and analysis of gametophyte defects were conducted. Complementation lines obtained by the native promoter and GCP4-coded CDS gene sequence fused with GFP reduced the numbers of lethal phenotypes of the gcp4 mutant. The localization of GCP4 in the gametophyte was detected in cytoplasm around nuclei and in vicinity of plasma membrane of pollen grains, and also detected in full cytoplasm and around the nuclei of ovules in complementation line. Thus, it was established that GCP4 influences the functionality of gametophytes during gametophyte development.
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References
Bahtz R, Seidler J, Arnold M, Haselmann-Weiss U, Antony C, Lehmann WD, Hoffmann I (2012) GCP6 is a substrate of Plk4 and required for centriole duplication. J Cell Sci 125:486–496. https://doi.org/10.1242/jcs.093930
Binarová P, Cenklová V, Procházková J, Doskočilová A, Volc J, Vrlík M, Bögre L (2006) γ-Tubulin is essential for acentrosomal microtubule nucleation and coordination of late mitotic events in Arabidopsis. Plant Cell 18:1199–1212. https://doi.org/10.1105/tpc.105.038364
Chen JWC, Chen ZA, Rogala KB, Metz J, Deane CM, Rappsilber J, Wakefield JG (2017) Cross-linking mass spectrometry identifies new interfaces of augmin required to localise the γ-tubulin ring complex to the mitotic spindle. Biology Open 6:654–663. https://doi.org/10.1242/bio.022905
Cota RR, Teixidó-Travesa N, Ezquerra A, Eibes S, Lacasa C, Roig J, Lüders J (2017) MZT1 regulates microtubule nucleation by linking γTuRC assembly to adapter-mediated targeting and activation. J Cell Sci 130:406–419. https://doi.org/10.1242/jcs.195321
Eady C, Lindsey K, Twell D (1995) The significance of microspore division and division symmetry for vegetative cell-specific transcription and generative cell differentiation. Plant Cell 7:65–74. https://doi.org/10.1105/tpc.7.1.65
Gibalová A, Reňák D, Matczuk K, Dupl’áková N, Cháb D, Twell D, Honys D (2009) AtbZIP34 is required for Arabidopsis pollen wall patterning and the control of several metabolic pathways in developing pollen. Plant Mol Biol 70:581–601. https://doi.org/10.1007/s11103-009-9493-y
Guevara-García A, López-Bucio J, Herrera-Estrella L (1999) The mannopine synthase promoter contains vectorial cis-regulatory elements that act as enhancers and silencers. MGG 262:608–617. https://doi.org/10.1007/s004380051123
Guillet V, Knibiehler M, Gregory-Pauron L, Remy MH, Chemin C, Raynaud-Messina B, Bon C, Kollman JM, Agard DA, Merdes A, Mourey L (2011) Crystal structure of gamma-tubulin complex protein GCP4 provides insight into microtubule nucleation. Nat Struct Mol Biol 18:915–919. https://doi.org/10.1038/nsmb.2083
Hamada T (2014) Microtubule organization and microtubule-associated proteins in plant cells. International Review of Cell & Molecular Biology 312:1–52. https://doi.org/10.1016/B978-0-12-800178-3.00001-4
Ho CM, Hotta T, Kong Z, Zeng CJ, Sun J, Lee YR, Liu B (2011) Augmin plays a critical role in organizing the spindle and phragmoplast microtubule arrays in Arabidopsis. Plant Cell 23:2606–2618. https://doi.org/10.1105/tpc.111.086892
Hotta T, Kong Z, Ho CM, Zeng CJ, Horio T, Fong S, Vuong T, Lee YR, Liu B (2012) Characterization of the Arabidopsis augmin complex uncovers its critical function in the assembly of the acentrosomal spindle and phragmoplast microtubule arrays. Plant Cell 24:1494–1509. https://doi.org/10.1105/tpc.112.096610
Izumi Nanae FK, Izumi S, Kikuchi A (2008) GSK-3β regulates proper mitotic spindle formation in cooperation with a component of the γ-tubulin ring complex, GCP5. J Biol Chem 283:12981–12991. https://doi.org/10.1074/jbc.M710282200
Janski N, Masoud K, Batzenschlager M, Herzog E, Evrard JL, Houlne G, Bourge M, Chaboute ME, Schmit AC (2012) The GCP3-interacting proteins GIP1 and GIP2 are required for gamma-tubulin complex protein localization, spindle integrity, and chromosomal stability. Plant Cell 24:1171–1187. https://doi.org/10.1105/tpc.111.094904
Kollman JM, Merdes A, Mourey L, Agard DA (2011) Microtubule nucleation by γ-tubulin complexes. Nat Rev Mol Cell Biol 12:709–721. https://doi.org/10.1038/nrm3209
Kong Z, Hotta T, Lee YR, Horio T, Liu B (2010) The γ-tubulin complex protein GCP4 is required for organizing functional microtubule arrays in Arabidopsis thaliana. Plant Cell 22:191–204. https://doi.org/10.1105/tpc.109.071191
Lee Y-RJ, Liu B (2019) Microtubule nucleation for the assembly of acentrosomal microtubule arrays in plant cells. New Phytol 222:1705–1718. https://doi.org/10.1111/nph.15705
Li LX, Liao HZ, Jiang LX, Tan Q, Ye D, Zhang XQ (2018) Arabidopsis thaliana NOP10 is required for gametophyte formation. JIPB 60:723–736. https://doi.org/10.1111/jipb.12652
Lin T-C, Neuner A, Schiebel E (2014) Targeting of γ-tubulin complexes to microtubule organizing centers: conservation and divergence. Trends Cell Biol 25:1–12. https://doi.org/10.1016/j.tcb.2014.12.002
Liu CM, Meinke DW (1998) The titan mutants of Arabidopsis are disrupted in mitosis and cell cycle control during seed development. Plant J 16:21–31. https://doi.org/10.1046/j.1365-313x.1998.00268.x
Liu B, Ho C-MK, Lee Y-RJ (2011) Microtubule reorganization during mitosis and cytokinesis: lessons learned from developing microgametophytes in Arabidopsis thaliana. Front Plant Sci 2:27. https://doi.org/10.3389/fpls.2011.00027
Liu T, Tian J, Wang G, Yu Y, Wang C, Ma Y, Zhang X, Xia G, Liu B, Kong Z (2014) Augmin triggers microtubule-dependent microtubule nucleation in interphase plant cells. Curr Biol 24:2708–2713. https://doi.org/10.1016/j.cub.2014.09.053
Ma D, Chen H, Han R (2017) Comparison of microtubule organization in Arabidopsis thaliana TUB-GFP and MBD-GFP mutants exposed to UV-B radiation. Journal of Agricultural Sciences 23:328–334. https://doi.org/10.15832/ankutbd.447642
Miao H, Guo R, Chen J, Wang Q, Lee Y-RJ, Liu B (2019) The γ-tubulin complex protein GCP6 is crucial for spindle morphogenesis but not essential for microtubule reorganization in Arabidopsis. Proc Natl Acad Sci 116:27115–27123. https://doi.org/10.1073/pnas.1912240116
Nakamura M, Hashimoto T (2009) A mutation in the Arabidopsis γ-tubulin-containing complex causes helical growth and abnormal microtubule branching. J Cell Sci 122:2208–2217. https://doi.org/10.1242/jcs.044131
Nakamura M, Yagi N, Kato T, Fujita S, Kawashima N, Ehrhardt DW, Hashimoto T (2012) Arabidopsis GCP3-interacting protein 1/MOZART 1 is an integral component of the gamma-tubulin-containing microtubule nucleating complex. Plant J 71:216–225. https://doi.org/10.1111/j.1365-313X.2012.04988.x
Oh SA, Pal MD, Park SK, Johnson JA, Twell D (2010) The tobacco MAP215/Dis1-family protein TMBP200 is required for the functional organization of microtubule arrays during male germline establishment. J Exp Bot 61:969–981. https://doi.org/10.1093/jxb/erp367
Oh SA, Jeon J, Park HJ, Grini PE, Twell D, Park SK (2016) Analysis of gemini pollen 3 mutant suggests a broad function of AUGMIN in microtubule organization during sexual reproduction in Arabidopsis. Plant J 87:188–201. https://doi.org/10.1111/tpj.13192
Pan Y, Li J, Jiao L, Li C, Zhu D, Yu J (2016) A non-specific Setaria italica lipid transfer protein gene plays a critical role under abiotic stress. Front Plant Sci 7:1752. https://doi.org/10.3389/fpls.2016.01752
Park SK, Howden R, Twell D (1998) The Arabidopsis thaliana gametophytic mutation gemini pollen1 disrupts microspore polarity, division asymmetry and pollen cell fate. Development 125:3789–3799
Pastuglia M, Azimzadeh J, Goussot M, Camilleri C, Belcram K, Evrard JL, Schmit AC, Guerche P, Bouchez D (2006) γ-Tubulin is essential for microtubule organization and development in Arabidopsis. Plant Cell 18:1412–1425. https://doi.org/10.1105/tpc.105.039644
Petry S, Vale RD (2015) Microtubule nucleation at the centrosome and beyond. Nat Cell Biol 17:1089–1093. https://doi.org/10.1038/ncb3220
Roostalu J, Surrey T (2017) Microtubule nucleation: beyond the template. Nat Rev Mol Cell Bio 18:702–710. https://doi.org/10.1038/nrm.2017.75
Smyth DR, Bowman JL, Meyerowitz EM (1990) Early flower development in Arabidopsis. Plant Cell 2:755–767. https://doi.org/10.1105/tpc.2.8.755
Struk S, Dhonukshe P (2014) MAPs: cellular navigators for microtubule array orientations in Arabidopsis. Plant Cell Rep 33:1–21. https://doi.org/10.1007/s00299-013-1486-2
Sulimenko V, Hájková Z, Klebanovych A, Dráber P (2017) Regulation of microtubule nucleation mediated by γ-tubulin complexes. Protoplasma 254:1187–1199. https://doi.org/10.1007/s00709-016-1070-z
Walia A, Nakamura M, Moss D, Kirik V, Hashimoto T, Ehrhardt DW (2014) GCP-WD mediates gamma-TuRC recruitment and the geometry of microtubule nucleation in interphase arrays of Arabidopsis. Curr Biol 24:2548–2555. https://doi.org/10.1016/j.cub.2014.09.013
Wang Q (2014) Mechanism analysis of the function of γ-tubulin complex protein 5 during Arabidopsis embryogenesis. Chinese Academy of Sciences
Wieczorek M, Bechstedt S, Chaaban S, Brouhard GJ (2015) Microtubule-associated proteins control the kinetics of microtubule nucleation. Nat Cell Biol 17:907–916. https://doi.org/10.1038/ncb3188
Yi P, Goshima G (2018) Microtubule nucleation and organization without centrosomes. Curr Opin Plant Biol 46:1–7. https://doi.org/10.1016/j.pbi.2018.06.004
Zeng CJ, Lee YR, Liu B (2009) The WD40 repeat protein NEDD1 functions in microtubule organization during cell division in Arabidopsis thaliana. Plant Cell 21:1129–1140. https://doi.org/10.1105/tpc.109.065953
Zhang X, Henriques R, Lin S, Niu Q, Chua N (2006) Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. Nat Protoc 1:641–646. https://doi.org/10.1038/nprot.2006.97
Acknowledgments
We thank Dr. Jinlin Feng (Shanxi Normal University) for critically reading the manuscript. We thank Professor Dongtao Ren (China Agricultural University) for kindly providing the pSuper1300-GFP vector.
Funding
This work was supported by a grant from 2018 Graduate Science and Technology Innovation of Shanxi Normal University (01053012) and 2019 Innovation Project of Postgraduate Education of Shanxi Province (2019BY090).
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Ma, D., Gao, L. & Han, R. Effects of the protein GCP4 on gametophyte development in Arabidopsis thaliana. Protoplasma 258, 483–493 (2021). https://doi.org/10.1007/s00709-020-01520-1
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DOI: https://doi.org/10.1007/s00709-020-01520-1