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Imaging the actin cytoskeleton in growing pollen tubes

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Abstract

Given the importance of the actin cytoskeleton to pollen tube growth, we have attempted to decipher its structure, organization and dynamic changes in living, growing pollen tubes of Nicotiana tabacum and Lilium formosanum, using three different GFP-labeled actin-binding domains. Because the intricate structure of the actin cytoskeleton in rapidly frozen pollen tubes was recently resolved, we now have a clear standard against which to compare the quality of labeling produced by these GFP-labeled probes. While GFP-talin, GFP-ADF and GFP-fimbrin show various aspects of the actin cytoskeleton structure, each marker produces a characteristic pattern of labeling, and none reveals the entire spectrum of actin. Whereas GFP-ADF, and to a lesser extent GFP-talin, label the fringe of actin in the apex, no similar structure is observed with GFP-fimbrin. Further, GFP-ADF only occasionally labels actin cables in the shank of the pollen tube, whereas GFP-fimbrin labels an abundance of fine filaments in this region, and GFP-talin bundles actin into a central cable in the core of the pollen tube surrounded by a few finer elements. High levels of expression of GFP-talin and GFP-fimbrin frequently cause structural rearrangements of the actin cytoskeleton of pollen tubes, and inhibit tip growth in a dose dependent manner. Most notably, GFP-talin results in thick cortical hoops of actin, transverse to the axis of growth, and GFP-fimbrin causes actin filaments to aggregate. Aberrations are seldom seen in pollen tubes expressing GFP-ADF. Although these markers are valuable tools to study the structure of the actin cytoskeleton of growing pollen tubes, given their ability to cause aberrations and to block pollen tube growth, we urge caution in their use.

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Acknowledgements

We thank Nam-Hai Chua, Zhenbiao Yang, Alice Cheung and Hen-ming Wu for providing us with constructs, and the Gloeckner Company for the donation of L. longiflorum bulbs. We thank Alice Cheung for the contribution of Fig. 6a and b. This research was supported by the National Science Foundation grant Nos. MCB-0077599 and MCB-0516852 to PKH and by the EU Research Training Network TIPNET (Project HPRN-CT-2002–00265), from Brussels, Belgium, to BV. We acknowledge the National Science Foundation Grant No. BBS 8714235 that supports the UMASS Central Microscopy Facility, where all confocal images were acquired. Finally, we acknowledge the David Delisle Research Grant, of the Plant Biology Graduate Program at UMASS, for its support.

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  1. Kathleen L. Wilsen

    Present address: Biological Sciences, University of Northern Colorado, Ross Hall 2480, Greeley, CO, 80639, USA

Authors and Affiliations

  1. Plant Biology Program, University of Massachusetts, 221 Morrill Science Center South, Amherst, MA, 01003-9297, USA

    Kathleen L. Wilsen, Alenka Lovy-Wheeler & Peter K. Hepler

  2. Institute of Botany, University of Bonn, Kirschallee 1, Bonn, Germany

    Boris Voigt & Diedrik Menzel

  3. Biology Department, University of Massachusetts, 221 Morrill Science Center South, Amherst, MA, 01003, USA

    Joseph G. Kunkel

Authors
  1. Kathleen L. Wilsen
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  2. Alenka Lovy-Wheeler
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  5. Joseph G. Kunkel
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  6. Peter K. Hepler
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Correspondence to Peter K. Hepler.

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Communicated by R. Russell

Financial Source: National Science Foundation grant Nos. MCB-0077599 and MCB-0516852 to PKH EU Research Training Network TIPNET (project HPRN-CT-2002-00265), Brussels, Belgium, to BV

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Wilsen, K.L., Lovy-Wheeler, A., Voigt, B. et al. Imaging the actin cytoskeleton in growing pollen tubes. Sex Plant Reprod 19, 51–62 (2006). https://doi.org/10.1007/s00497-006-0021-9

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