Skip to main content
SpringerLink
Log in

Microtubules and microfilaments coordinate to direct a fountain streaming pattern in elongating conifer pollen tube tips

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

This study investigates how microtubules and microfilaments control organelle motility within the tips of conifer pollen tubes. Organelles in the 30-μm-long clear zone at the tip of Picea abies (L.) Karst. (Pinaceae) pollen tubes move in a fountain pattern. Within the center of the tube, organelles move into the tip along clearly defined paths, move randomly at the apex, and then move away from the tip beneath the plasma membrane. This pattern coincides with microtubule and microfilament organization and is the opposite of the reverse fountain seen in angiosperm pollen tubes. Application of latrunculin B, which disrupts microfilaments, completely stops growth and reduces organelle motility to Brownian motion. The clear zone at the tip remains intact but fills with thin tubules of endoplasmic reticulum. Applications of amiprophosmethyl, propyzamide or oryzalin, which all disrupt microtubules, stop growth, alter organelle motility within the tip, and alter the organization of actin microfilaments. Amiprophosmethyl inhibits organelle streaming and collapses the clear zone of vesicles at the extreme tip together with the disruption of microfilaments leading into the tip, leaving the plasma membrane intact. Propyzamide and oryzalin cause the accumulation of membrane tubules or vacuoles in the tip that reverse direction and stream in a reverse fountain. The microtubule disruption caused by propyzamide and oryzalin also reorganizes microfilaments from a fibrillar network into pronounced bundles in the tip cytoplasm. We conclude that microtubules control the positioning of organelles into and within the tip and influence the direction of streaming by mediating microfilament organization.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Figs. 1–4 Fig. 1 Fig. 2 Fig. 3 Fig. 4
Figs. 5–10 Fig. 5 Fig. 6 Fig. 7 Fig. 8a, b Fig. 9 Fig. 10

Similar content being viewed by others

Abbreviations

APM :

Amiprophosmethyl

FITC :

Fluorescein isothiocyanate

LATB :

Latrunculin B

References

  • Anderhag P, Hepler PK, Lazzaro MD (2000) Microtubules and microfilaments are both responsible for pollen tube elongation in the conifer Picea abies (Norway spruce). Protoplasma 214:141–157

    Google Scholar 

  • Andersland JM, Fisher DD, Wymer CL, Cyr RJ, Parthasarathy MV (1994) Characterization of a monoclonal antibody prepared against plant actin. Cell Motil Cytoskel 29:339–344

    CAS  Google Scholar 

  • Åström H, Sorri O, Raudaskoski M (1995) Role of microtubules in the movement of the vegetative nucleus and generative cell in tobacco pollen tubes. Sex Plant Reprod 8:61–69

    Google Scholar 

  • Brandizzi F, Snapp EL, Roberts AG, Lippincott-Schwartz J, Hawes C (2002) Membrane protein transport between the endoplasmic reticulum and the Golgi in tobacco leaves is energy dependent but cytoskeleton independent: evidence from selective photobleaching. Plant Cell 14:1293–1309

    Article  CAS  PubMed  Google Scholar 

  • Cai G, Romagnoli S, Moscatelli A, Ovidi E, Gambellini G, Tiezzi A, Cresti M (2000) Identification and characterization of a novel microtubule based motor associated with membranous organelles in tobacco pollen tubes. Plant Cell 12:1719–1736

    CAS  PubMed  Google Scholar 

  • Chen CY, Wong EI, Vidali L, Estavillo A, Hepler PK, Wu HM, Cheung AY (2002) The regulation of actin organization by actin-depolymerizing factor in elongating pollen tubes. Plant Cell 14:2175–2190

    Article  CAS  PubMed  Google Scholar 

  • Chen CY, Cheung AY, Wu HM (2003) Actin-depolymerizing factor mediates Rac/Rop GTPase regulated pollen tube growth. Plant Cell 15:237–249

    Google Scholar 

  • Cole L, Davies D, Hyde GJ, Ashford AE (2000) Brefeldin A affects growth, endoplasmic reticulum, Golgi bodies, tubular vacuole system, and secretory pathway in Pisolithus tinctorius. Fungal Genet Biol 29:95–106

    Article  CAS  PubMed  Google Scholar 

  • Dawkins MD, Owens JN (1993) In vitro and in vivo pollen hydration, germination, and pollen tube growth in white spruce, Picea glauca (Moench) Voss. Int J Plant Sci 154:506–521

    Article  Google Scholar 

  • de Win AHN, Knuiman B, Pierson ES, Geurts H, Kengen HMP, Derksen J (1996) Development and cellular organization of Pinus sylvestris pollen tubes. Sex Plant Reprod 9:93–101

    Article  Google Scholar 

  • Feijo JA, Sainhas J, Holdaway-Clarke T, Cordeiro MS, Kunkel JG, Hepler PK (1999) Growing pollen tubes possess a constitutive alkaline band in the clear zone and a growth dependent acidic tip. J Cell Biol 144:483–496

    Article  PubMed  Google Scholar 

  • Gibbon BC, Kovar DR, Staiger CJ (1999) Latrunculin B has different effects on pollen germination and tube growth. Plant Cell 11:2349–2363

    CAS  PubMed  Google Scholar 

  • Hepler PK, Vidali L, Cheung AY (2001) Polarized cell growth in higher plants. Annu Rev Cell Dev Biol 17:159–187

    CAS  PubMed  Google Scholar 

  • Heslop-Harrison J, Heslop-Harrison Y (1989) Myosin associated with the surfaces of organelles, vegetative nuclei and generative cells in angiosperm pollen grains and tubes. J Cell Sci 94:319–325

    Google Scholar 

  • Heslop-Harrison J, Heslop-Harrison Y, Cresti M, Tiezzi A, Moscatelli A (1988) Cytoskeletal elements, cell shaping and movement in the angiosperm pollen tube. J Cell Sci 91:49–60

    Google Scholar 

  • Hoffman JC, Vaughn KC (1994) Mitotic disruptors act by a single mechanism but vary in efficacy. Protoplasma 179:16–25

    CAS  Google Scholar 

  • Iwai S, Ishiji A, Mabuchi I, Sutoh K (2003) A novel actin-bundling kinesin-related protein from Dictyostelium discoideum. J Biol Chem DOI 10.1074/jbc.M308022200

  • Kadota A, Yoshizaki N, Wada M (1999) Cytoskeletal changes during resumption of tip growth in non-growing protonema cells of the fern Adiantum capillus-veneris L. Protoplasma 207:195–202

    Google Scholar 

  • Krishnakumar S, Oppenheimer DG (1999) Extragenic suppressors of the Arabidopsis zwi-3 mutation identify new genes that function in trichome branch formation and pollen tube growth. Development 126:3079–3088

    CAS  PubMed  Google Scholar 

  • Lancelle SA, Hepler PK (1992) Ultrastructure of freeze substituted pollen tubes of Lilium longiflorum. Protoplasma 167:215–230

    Google Scholar 

  • Lazzaro MD (1996) The actin microfilament network within elongating pollen tubes of the gymnosperm Picea abies (Norway spruce). Protoplasma 194:186–194

    CAS  Google Scholar 

  • Lazzaro MD (1999) Microtubule organization in germinated pollen of the conifer Picea abies (Norway spruce—Pinaceae). Am J Bot 86:759–766

    PubMed  Google Scholar 

  • Lazzaro MD, Donohue JM, Soodavar FM (2003) Disruption of cellulose synthesis by isoxaben causes tip swelling and disorganizes cortical microtubules in elongating conifer pollen tubes. Protoplasma 220:201–207

    Article  CAS  PubMed  Google Scholar 

  • Miller DD, Lancelle SA, Hepler PK (1996) Actin microfilaments do not form a dense meshwork in Lilium longiflorum pollen tubes. Protoplasma 195:123–132

    Google Scholar 

  • Parton RM, Fischer-Parton S, Watahiki MK, Trewavas AJ (2001) Dynamics of the apical vesicle accumulation and the rate of growth are related in individual pollen tubes. J Cell Sci 114:2685–2695

    CAS  PubMed  Google Scholar 

  • Pierson ES, Derksen J, Traas JA (1986) Organization of microfilaments and microtubules in pollen tubes grown in vitro and in vivo in various angiosperms. Eur J Cell Biol 41:14–18

    Google Scholar 

  • Pierson ES, Miller DD, Callaham DA, van Aken J, Hackett G, Hepler PK (1996) Tip-localized calcium entry fluctuates during pollen tube growth. Dev Biol 174:160–173

    Article  CAS  PubMed  Google Scholar 

  • Reddy ASN (2001) Molecular motors and their functions in plants. Int Rev Cytol 204:97–178

    CAS  PubMed  Google Scholar 

  • Romagnoli S, Cai G, Cresti M (2003) In vitro assays demonstrate that pollen tube organelles use kinesin related motor proteins to move along microtubules. Plant Cell 15:251–269

    Google Scholar 

  • Runions CJ, Owens JN (1999) Sexual reproduction of interior spruce (Pinaceae) II: fertilization to early embryo formation. Int J Plant Sci 160:641–652

    Article  Google Scholar 

  • Schwuchow J, Sack FD, Hartmann E (1990) Microtubule distribution in gravitropic protonemata of the moss Ceratodon. Protoplasma 159:60–69

    CAS  PubMed  Google Scholar 

  • Taylor LP, Hepler PK (1997) Pollen germination and tube growth. Annu Rev Plant Physiol Plant Mol Biol 48:4161–491

    Article  Google Scholar 

  • Tominaga M, Morita K, Sonobe S, Yokota E, Shimmen T (1997) Microtubules regulate the organization of actin filaments at the cortical region in root hairs of Hydrocharis. Protoplasma 199:83–92

    CAS  Google Scholar 

  • Vidali L, McKenna ST, Hepler PK (2001) Actin polymerization is essential for pollen tube growth. Mol Biol Cell 12:2534–2545

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Kevin Vaughn for insightful suggestions and Richard Cyr for the pea root actin antibody. Dow AgroSciences donated propyzamide and oryzalin, and Bayer Corporation donated APM. This work was supported by research grants from the Department of Biology, College of Charleston and by NIH grant number RR-16461-01 from the BRIN Program of the National Center for Research Resources. C.D. Justus, P. Anderhag and J.L. Goins were all undergraduate students.

Author information

Authors and Affiliations

  1. Department of Biology, College of Charleston, Charleston, SC 29424, USA

    Charles D. Justus, Per Anderhag, Jeanne L. Goins & Mark D. Lazzaro

Authors
  1. Charles D. Justus
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Per Anderhag
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jeanne L. Goins
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mark D. Lazzaro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mark D. Lazzaro.

Electronic Supplementary Material

Supplemental videos of pollen tubes

Videos are in Quicktime format. Download the free Quicktime plugin. You can contact the authors at lazzarom@cofc.edu or see the Lazzaro Lab website at www.cofc.edu/~lazzaro

(QuickTime Movie 2.6 MB)

(QuickTime Movie 1.7 MB)

(QuickTime Movie 3.1 MB)

(QuickTime Movie 5.3 MB)

(QuickTime Movie 4.5 MB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Justus, C.D., Anderhag, P., Goins, J.L. et al. Microtubules and microfilaments coordinate to direct a fountain streaming pattern in elongating conifer pollen tube tips. Planta 219, 103–109 (2004). https://doi.org/10.1007/s00425-003-1193-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-003-1193-2

Keywords

Search

Navigation