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The speed of mitochondrial movement is regulated by the cytoskeleton and myosin in Picea wilsonii pollen tubes

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Abstract

Strategic control of mitochondrial movements and cellular distribution is essential for correct cell function and survival. However, despite being a vital process, mitochondrial movement in plant cells is a poorly documented phenomenon. To investigate the roles of actin filaments and microtubules on mitochondrial movements, Picea wilsonii pollen tubes were treated with two microtubule-disrupting drugs, two actin-disrupting drugs and a myosin inhibitor. Following these treatments, mitochondrial movements were characterized by multiangle evanescent wave microscopy and laser-scanning confocal microscopy. The results showed that individual mitochondria underwent three classes of linear movement: high-speed movement (instantaneous velocities >5.0 μm/s), low-speed movement (instantaneous velocities <5.0 μm/s) and variable-speed movement (instantaneous velocities ranging from 0.16 to 10.35 μm/s). 10 nM latrunculin B induced fragmentation of actin filaments and completely inhibited mitochondrial vectorial movement. Jasplakinolide treatment induced a 28% reduction in chondriome motility, and dramatically inhibition of high-speed and variable-speed movements. Treatment with 2,3-butanedione 2-monoxime caused a 61% reduction of chondriome motility, and the complete inhibition of high-speed and low-speed movements. In contrast to actin-disrupting drugs, microtubule-disrupting drugs caused mild effects on mitochondrial movement. Taxol increased the speed of mitochondrial movement in cortical cytoplasm. Oryzalin induced curved mitochondrial trajectories with similar velocities as in the control pollen tubes. These results suggest that mitochondrial movement at low speeds in pollen tubes is driven by myosin, while high-speed and variable-speed movements are powered both by actin filament dynamics and myosin. In addition, microtubule dynamics has profound effects on mitochondrial velocity, trajectory and positioning via its role in directing the arrangement of actin filaments.

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Abbreviations

EWM:

Evanescent wave microscopy

LatB:

Latrunculin B

Jas:

Jasplakinolide

BDM:

2,3-Butanedione 2-monoxime

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Acknowledgments

This work was supported by grants from the Knowledge Innovation Program of the Chinese Academy of Sciences (KJCX2-YW-L08) and A Hundred Talents Programme from the Chinese Academy of Sciences, and by the European Union Research Training Network TIPNET (project HPRN-CT-2002-00265), by Grant Agency APVT (grant no. APVT-51-002302). DCL is supported by the University of Saskatchewan and research grants from the Natural Sciences and Engineering Research Council of Canada, and the Canada Foundation for Innovation. Visits to China were supported by the Royal Society of Edinburgh International Exchange Programme.

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Authors and Affiliations

  1. Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Xiangshan, 100093, Beijing, China

    Maozhong Zheng, Qinli Wang, Xiaohua Wang, Feng Wang, Tong Chen & Jinxing Lin

  2. Graduate School of Chinese Academy of Sciences, 100049, Beijing, China

    Maozhong Zheng, Xiaohua Wang & Feng Wang

  3. Institute of Biophysics, Chinese Academy of Sciences, 100101, Beijing, China

    Yan Teng

  4. Department of Plant Cell Biology, Institute of Cellular and Molecular Botany, Rheinische Friedrich-Wilhelms-University Bonn, Kirschallee 1, 53115, Bonn, Germany

    Jozef Šamaj

  5. Faculty of Science, Palacký University Olomouc, 783 71, Olomouc, Czech Republic

    Jozef Šamaj

  6. Department of Biology, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5E2, Canada

    David C. Logan

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  1. Maozhong Zheng
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  2. Qinli Wang
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  3. Yan Teng
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  4. Xiaohua Wang
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  5. Feng Wang
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  7. Jozef Šamaj
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  8. Jinxing Lin
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Correspondence to Jinxing Lin or David C. Logan.

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Supplemental Fig. S1

Effect of taxol and oryzalin on microtubules in Picea wilsonii pollen tubes. Samples were chemically fixed and immunolabeled with anti-tubulin antibody, then visualized by confocal microscopy. a In untreated pollen tubes microtubules were found throughout the tube, especially as a dense network in the cell cortex. b A pollen tube treated with 100 μM oryzalin for 10 min, showing disruption to the microtubule cytoskeleton. c A pollen tube treated with 5 μM taxol for 10 min, showing thick microtubules. Bars = 20 μm. (TIFF 586 kb)

425_2009_1086_MOESM2_ESM.avi

Supplementary video 1: Video sequence showing mitochondrial movement in a pollen tube treated with 0.5% (w/v) DMSO for 10 min. Images were collected at 5-s intervals by confocal microscopy and played back at 10 frames per second (fps). Total elapsed time is 260 s. (AVI 5188 kb)

Supplementary video 2: Video sequence showing mitochondrial movement in a pollen tube treated with 0.5% (w/v) ethanol for 10 min. Images were collected at 5-s intervals by confocal microscopy and played back at 10 frames per second (fps). Total elapsed time is 325s. (AVI 4846 kb)

425_2009_1086_MOESM4_ESM.avi

Supplementary video 3: Video sequence showing the enhanced mitochondrial movement in the cortical cytoplasm in a pollen tube treated with 5 μM taxol for 10 min. Images were collected at 5-s intervals by confocal microscopy and played back at 5 frames per second (fps). Total elapsed time is 315 s. (AVI 9953 kb)

425_2009_1086_MOESM5_ESM.avi

Supplemental video 4: Video sequence showing mitochondrial movement in an untreated Picea wilsonii pollen tube. Images were collected at 5-s intervals by confocal microscopy and played back at 10 frames per second (fps). Total elapsed time is 500 s. (AVI 6236 kb)

425_2009_1086_MOESM6_ESM.avi

Supplementary video 5: Video sequence showing various mitochondrial movements in an untreated Picea wilsonii pollen tube. Images were collected at 200 ms intervals using multiangle evanescent wave microscopy and played back at 20 frames per second (fps). Total elapsed time is 17.6 s. (AVI 9648 kb)

Supplemental video 6: Video sequence showing modified trajectories of mitochondrial movements in a pollen tube treated with 100 μM oryzalin for 10 min. Images were collected at 200 ms intervals using multiangle evanescent wave microscopy and played back at 20 frames per second (fps). Total elapsed time is 10 s. (AVI 10673 kb)

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Zheng, M., Wang, Q., Teng, Y. et al. The speed of mitochondrial movement is regulated by the cytoskeleton and myosin in Picea wilsonii pollen tubes. Planta 231, 779–791 (2010). https://doi.org/10.1007/s00425-009-1086-0

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