Abstract
Pollen tubes require functional mitochondria in order to achieve fast and sustained growth. In addition, cell wall expansion requires a calcium gradient in the tube apex formed by a dedicated array of calcium pumps and channels. Most studies have traditionally focused on the molecular aspects of calcium interactions and transport across the pollen tube plasmalemma. However, calcium transients across mitochondrial membranes from pollen tubes are beginning to be studied. Here, we report the presence of a ruthenium red-sensitive mitochondrial calcium uniporter-like activity in tobacco pollen tubes with functional oxidative phosphorylation. The present study provides a framework to measure in situ specifics of mitochondrial transport and respiration in pollen tubes from different plants. The relevance of a mitochondrial calcium uniporter for pollen tube growth is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- MCU:
-
mitochondrial calcium uniporter
- AtMCU1:
-
Arabidopsis thaliana mitochondrial calcium uniporter 1
- AtMCU2:
-
Arabidopsis thaliana mitochondrial calcium uniporter 2
- MICU:
-
EF-hand MCU regulator
- MPT:
-
mitochondrial permeability transition
- PG:
-
pollen germination
- PTA:
-
pollen tube assay
- RuR:
-
ruthenium red
- ROS:
-
reactive oxygen species
References
Baines CP, Gutierrez-Aguilar M (2018) The still uncertain identity of the channel-forming unit(s) of the mitochondrial permeability transition pore. Cell Calcium 73:121–130. https://doi.org/10.1016/j.ceca.2018.05.003
Baughman JM, Perocchi F, Girgis HS, Plovanich M, Belcher-Timme CA, Sancak Y, Bao XR, Strittmatter L, Goldberger O, Bogorad RL, Koteliansky V, Mootha VK (2011) Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 476:341–345. https://doi.org/10.1038/nature10234
Bosch M, Cheung AY, Hepler PK (2005) Pectin methylesterase, a regulator of pollen tube growth. Plant Physiol 138:1334–1346. https://doi.org/10.1104/pp.105.059865
Bosch M, Hepler PK (2005) Pectin methylesterases and pectin dynamics in pollen tubes. Plant Cell 17:3219–3226. https://doi.org/10.1105/tpc.105.037473
Bosch M, Hepler PK (2006) Silencing of the tobacco pollen pectin methylesterase NtPPME1 results in retarded in vivo pollen tube growth. Planta 223:736–745. https://doi.org/10.1007/s00425-005-0131-x
Braccini I, Perez S (2001) Molecular basis of C(2+)-induced gelation in alginates and pectins: the egg-box model revisited. Biomacromolecules 2:1089–1096
Claros MG (1995) MitoProt, a Macintosh application for studying mitochondrial proteins. Comput Appl Biosci 11:441–447
Collins S, Meyer T (2010) Cell biology: a sensor for calcium uptake. Nature 467:283. https://doi.org/10.1038/467283a
De Stefani D, Raffaello A, Teardo E, Szabo I, Rizzuto R (2011) A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature 476:336–340. https://doi.org/10.1038/nature10230
Deluca HF, Engstrom GW (1961) Calcium uptake by rat kidney mitochondria. Proc Natl Acad Sci U S A 47:1744–1750
Denton RM, McCormack JG (1980) On the role of the calcium transport cycle in heart and other mammalian mitochondria. FEBS Lett 119:1–8
Fiskum G (1985) Intracellular levels and distribution of Ca2+ in digitonin-permeabilized cells. Cell Calcium 6:25–37
He L, Li B, Lu X, Yuan L, Yang Y, Yuan Y, du J, Guo S (2015) The effect of exogenous calcium on mitochondria, respiratory metabolism enzymes and ion transport in cucumber roots under hypoxia. Sci Rep 5:11391. https://doi.org/10.1038/srep11391
Hepler PK, Kunkel JG, Rounds CM, Winship LJ (2012) Calcium entry into pollen tubes. Trends Plant Sci 17:32–38. https://doi.org/10.1016/j.tplants.2011.10.007
Heslop-Harrison J, Heslop-Harrison Y (1988) Some permeability properties of angiosperm pollen grains, pollen tubes and generative cells. Sex Plant Reprod 1:65–73. https://doi.org/10.1007/bf00189264
Iwano M, Entani T, Shiba H, Kakita M, Nagai T, Mizuno H, Miyawaki A, Shoji T, Kubo K, Isogai A, Takayama S (2009) Fine-tuning of the cytoplasmic Ca2+ concentration is essential for pollen tube growth. Plant Physiol 150:1322–1334. https://doi.org/10.1104/pp.109.139329
Kovacs-Bogdan E, Sancak Y, Kamer KJ, Plovanich M, Jambhekar A, Huber RJ, Myre MA, Blower MD, Mootha VK (2014) Reconstitution of the mitochondrial calcium uniporter in yeast. Proc Natl Acad Sci U S A 111:8985–8990. https://doi.org/10.1073/pnas.1400514111
Loro G, Drago I, Pozzan T, Schiavo FL, Zottini M, Costa A (2012) Targeting of Cameleons to various subcellular compartments reveals a strict cytoplasmic/mitochondrial Ca(2)(+) handling relationship in plant cells. Plant J 71:1–13. https://doi.org/10.1111/j.1365-313X.2012.04968.x
Loro G, Ruberti C, Zottini M, Costa A (2013) The D3cpv Cameleon reports Ca(2)(+) dynamics in plant mitochondria with similar kinetics of the YC3.6 Cameleon, but with a lower sensitivity. J Microsc 249:8–12. https://doi.org/10.1111/j.1365-2818.2012.03683.x
Lupas A, Van Dyke M, Stock J (1991) Predicting coiled coils from protein sequences. Science 252:1162–1164. https://doi.org/10.1126/science.252.5009.1162
Mammucari C, Raffaello A, Vecellio Reane D, Gherardi G, De Mario A, Rizzuto R (2018) Mitochondrial calcium uptake in organ physiology: from molecular mechanism to animal models Pflugers Arch https://doi.org/10.1007/s00424-018-2123-2
Oxenoid K, Dong Y, Cao C, Cui T, Sancak Y, Markhard AL, Grabarek Z, Kong L, Liu Z, Ouyang B, Cong Y, Mootha VK, Chou JJ (2016) Architecture of the mitochondrial calcium uniporter. Nature 533:269–273. https://doi.org/10.1038/nature17656
Pan X, Liu J, Nguyen T, Liu C, Sun J, Teng Y, Fergusson MM, Rovira II, Allen M, Springer DA, Aponte AM, Gucek M, Balaban RS, Murphy E, Finkel T (2013) The physiological role of mitochondrial calcium revealed by mice lacking the mitochondrial calcium uniporter. Nat Cell Biol 15:1464–1472. https://doi.org/10.1038/ncb2868
Penna E, Espino J, De Stefani D, Rizzuto R (2018) The MCU complex in cell death. Cell Calcium 69:73–80. https://doi.org/10.1016/j.ceca.2017.08.008
Perez-Vazquez V, Saavedra-Molina A, Uribe S (2003) In Saccharomyces cerevisiae, cations control the fate of the energy derived from oxidative metabolism through the opening and closing of the yeast mitochondrial unselective channel. J Bioenerg Biomembr 35:231–241
Perocchi F, Gohil VM, Girgis HS, Bao XR, McCombs JE, Palmer AE, Mootha VK (2010) MICU1 encodes a mitochondrial EF hand protein required for Ca(2+) uptake. Nature 467:291–296. https://doi.org/10.1038/nature09358
Qi Z, Stephens NR, Spalding EP (2006) Calcium entry mediated by GLR3.3, an Arabidopsis glutamate receptor with a broad agonist profile. Plant Physiol 142:963–971. https://doi.org/10.1104/pp.106.088989
Rounds CM, Hepler PK, Fuller SJ, Winship LJ (2010) Oscillatory growth in lily pollen tubes does not require aerobic energy metabolism. Plant Physiol 152:736–746. https://doi.org/10.1104/pp.109.150896
Rounds CM, Winship LJ, Hepler PK (2011) Pollen tube energetics: respiration, fermentation and the race to the ovule. AoB Plants 2011:plr019. https://doi.org/10.1093/aobpla/plr019
Schiott M, Romanowsky SM, Baekgaard L, Jakobsen MK, Palmgren MG, Harper JF (2004) A plant plasma membrane Ca2+ pump is required for normal pollen tube growth and fertilization. Proc Natl Acad Sci U S A 101:9502–9507. https://doi.org/10.1073/pnas.0401542101
Selles B, Michaud C, Xiong TC, Leblanc O, Ingouff M (2018) Arabidopsis pollen tube germination and growth depend on the mitochondrial calcium uniporter complex. New Phytol 219:58–65. https://doi.org/10.1111/nph.15189
Sommakia S, Houlihan PR, Deane SS, Simcox JA, Torres NS, Jeong MY, Winge DR, Villanueva CJ, Chaudhuri D (2017) Mitochondrial cardiomyopathies feature increased uptake and diminished efflux of mitochondrial calcium. J Mol Cell Cardiol 113:22–32. https://doi.org/10.1016/j.yjmcc.2017.09.009
Song LF, Zou JJ, Zhang WZ, Wu WH, Wang Y (2009) Ion transporters involved in pollen germination and pollen tube tip-growth. Plant Signal Behav 4:1193–1195
Sonnhammer EL, von Heijne G, Krogh A (1998) A hidden Markov model for predicting transmembrane helices in protein sequences. Proc Int Conf Intell Syst Mol Biol 6:175–182
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Teardo E, Carraretto L, Wagner S, Formentin E, Behera S, de Bortoli S, Larosa V, Fuchs P, Lo Schiavo F, Raffaello A, Rizzuto R, Costa A, Schwarzländer M, Szabò I (2017) Physiological characterization of a plant mitochondrial calcium uniporter in vitro and in vivo. Plant Physiol 173:1355–1370. https://doi.org/10.1104/pp.16.01359
Uribe S, Rangel P, Pardo JP (1992) Interactions of calcium with yeast mitochondria. Cell Calcium 13:211–217
Uribe-Carvajal S, Luevano-Martinez LA, Guerrero-Castillo S, Cabrera-Orefice A, Corona-de-la-Pena NA, Gutierrez-Aguilar M (2011) Mitochondrial Unselective Channels throughout the eukaryotic domain. Mitochondrion 11:382–390. https://doi.org/10.1016/j.mito.2011.02.004
von Stockum S, Basso E, Petronilli V, Sabatelli P, Forte MA, Bernardi P (2011) Properties of Ca(2+) transport in mitochondria of Drosophila melanogaster. J Biol Chem 286:41163–41170. https://doi.org/10.1074/jbc.M111.268375
Waese J, Fan J, Pasha A, Yu H, Fucile G, Shi R, Cumming M, Kelley LA, Sternberg MJ, Krishnakumar V, Ferlanti E, Miller J, Town C, Stuerzlinger W, Provart NJ (2017) ePlant: visualizing and exploring multiple levels of data for hypothesis generation in plant biology. Plant Cell 29:1806–1821. https://doi.org/10.1105/tpc.17.00073
Williams GS, Boyman L, Chikando AC, Khairallah RJ, Lederer WJ (2013) Mitochondrial calcium uptake. Proc Natl Acad Sci U S A 110:10479–10486. https://doi.org/10.1073/pnas.1300410110
Acknowledgements
This work was supported by grant UNAM-FQ-PAIP 5000-9171 (to M.G.-A.). We would like to thank Dr. Sobeida Sánchez-Nieto and Dr. Felipe Cruz-García for providing valuable material resources and advice for the completion of this study.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Handling Editor: Peter Nick
Electronic supplementary material
Figure S1
(A) Predicted Nicotiana tabacum MCU orthologues using Mus musculus MCU as template were found by a Basic Local Alignment Search Tool and aligned using Clustal Omega. Highly conserved residues encompassing the RuR- and calcium-binding domain are colored. Mitochondrial import probability (P) was assessed using Mitoprot and displayed after each predicted protein sequence segment alignment. (B) Mus musculus MICU1 was used as template for a Basic Local Alignment Search Tool. A phylogenetic tree was constructed using with the NCBI phylogenetic tree tool. (PNG 509 kb)
Rights and permissions
About this article
Cite this article
Flores-Herrera, C., Preciado-Linares, G., Gonzalez-Vizueth, I. et al. In situ assessment of mitochondrial calcium transport in tobacco pollen tubes. Protoplasma 256, 503–509 (2019). https://doi.org/10.1007/s00709-018-1316-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-018-1316-z