Skip to main content
Springer Nature Link
Log in

Sperm dimorphism in terms of nuclear shape and microtubule accumulation in Cyrtanthus mackenii

  • Original Article
  • Published:
Sexual Plant Reproduction Aims and scope Submit manuscript

Abstract

Pollen tubes of Cyrtanthus mackenii, a species with bicellular pollen, were cultured in vitro to investigate nuclear phase changes during generative cell division and male germ unit (MGU) formation, using flow cytometric analysis. Results revealed that sperm cells were formed after 12 h of culture. During sperm maturation, the nuclei of sperm cells were not associated with the vegetative nucleus (unassociated sperm cells; Sua) and became longer than those of sperm cells associated with the vegetative nucleus (Svn). These findings indicate that the pair of sperm cells in the C. mackenii MGU is dimorphic in terms of nuclear shape. Dimorphism coincides with anti-α-tubulin antibody immunofluorescence, which was higher in the Sua than in Svn. Following treatment with oryzalin, triggering microtubule depolymerization, differences between nuclear shapes in the two sperm nuclei disappeared, suggesting that microtubule accumulation between sperm cells in the MGU correlates with differences in the nuclear shape.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Cai G, Ovidi E, Romagnoli S, Vantard M, Cresti M, Tiezzi A (2005) Identification and characterization of plasma membrane proteins that bind to microtubules in pollen tubes and generative cells of tobacco. Plant Cell Physiol 46:563–578

    Article  CAS  PubMed  Google Scholar 

  • Chen SH, Liao JP, Kuang AX, Tian HQ (2006) Isolation of two populations of sperm cells from the pollen tube of Torenia fournieri. Plant Cell Rep 25:1138–1142

    Article  CAS  PubMed  Google Scholar 

  • Dumas C, Knox RB, McConchie CA, Russell SD (1984) Emerging physiological concepts in fertilization. What’s New Plant Physiol 15:17–20

    Google Scholar 

  • Gou X, Yuan T, Wei X, Russell SD (2009) Gene expression in the dimorphic sperm cells of Plumbago zeylanica: transcript profiling, diversity, and relationship to cell type. Plant J 60:33–47

    Article  CAS  PubMed  Google Scholar 

  • Hirano T, Hoshino Y (2009) Detection of changes in the nuclear phase and evaluation of male germ units by flow cytometry during in vitro pollen tube growth in Alstroemeria aurea. J Plant Res 122:225–234

    Article  PubMed  Google Scholar 

  • Hoshino Y, Scholten S, von Wiegen P, Lörz H, Kranz E (2004) Fertilization-induced changes in the microtubular architecture of the maize egg cell and zygote—an immunocytochemical approach adapted to single cells. Sex Plant Reprod 17:89–95

    Article  CAS  Google Scholar 

  • Kranz E, Hoshino Y, Okamoto T (2008) In vitro fertilization with isolation higher plant gametes. In: Suarez MF, Bozhkov PV (eds) Plant embryogenesis: method in molecular biology, vol 427. Humana Press, Totowa, NJ, pp 51–69

    Chapter  Google Scholar 

  • Lalanne E, Twell D (2002) Genetic control of male germ unit organization in Arabidopsis. Plant Physiol 129:865–875

    Article  CAS  PubMed  Google Scholar 

  • Lee YRJ, Li Y, Liu B (2007) Two Arabidopsis phragmoplast-associated kinesins play a critical role on cytokinesis during male gametogenesis. Plant Cell 19:2595–2605

    Article  CAS  PubMed  Google Scholar 

  • Mogensen HL (1992) The male germ unit: concept, composition, and significance. Intl Rev Cytol 140:129–147

    Article  Google Scholar 

  • Mori T, Kuroiwa H, Higashiyama T, Kuroiwa T (2003) Identification of higher plant GlsA, a putative morphogenesis factor of gametic cells. Biochem Biophys Res Commun 306:564–569

    Article  CAS  PubMed  Google Scholar 

  • Palevitz BA, Tiezzi A (1992) Organization, composition, and function of the generative cell and sperm cytoskeleton. Intl Rev Cytol 140:149–185

    Article  Google Scholar 

  • Russell SD (1984) Ultrastructure of the sperm of Plumbago zeylanica. II. Quantitative cytology and three-dimensional organization. Planta 162:385–391

    Article  Google Scholar 

  • Russell SD (1985) Preferential fertilization in Plumbago: ultrastructural evidence for gamete-level recognition in an angiosperm. Proc Natl Acad Sci USA 82:6129–6132

    Article  CAS  PubMed  Google Scholar 

  • Russell SD (1991) Isolation and characterization of sperm cells in flowering plants. Annu Rev Plant Physiol Plant Mol Biol 42:189–204

    Article  CAS  Google Scholar 

  • Russell SD, Cass DD (1981) Ultrastructure of the sperms of Plumbago zeylanica. 1. Cytology and association with vegetative nucleus. Protoplasma 107:85–107

    Article  Google Scholar 

  • Russell SD, Strout GW (2005) Microgametogenesis in Plumbago zeylanica (Plumbaginaceae). 2. Quantitative cell and organelle dynamics of the male reproductive cell lineage. Sex Plant Reprod 18:113–130

    Article  Google Scholar 

  • Russell SD, Rougier M, Dumas C (1990) Organization of the early post-fertilization megagametophyte of Populus deltoides ultrastructure and implications for male cytoplasmic transmission. Protoplasma 155:153–165

    Article  Google Scholar 

  • Saito C, Nagata N, Sakai A, Mori K, Kuroiwa H, Kuroiwa T (2002) Angiosperm species that produce sperm cell pairs or generative cells with polarized distribution of DNA-containing organelles. Sex Plant Reprod 15:167–178

    Article  CAS  Google Scholar 

  • Singh MB, Xu H, Bhalla PL, Zhang Z, Swoboda I, Russell SD (2002) Developmental expression of polyubiquitin genes and distribution of ubiquitinated proteins in generative and sperm cells. Sex Plant Reprod 14:325–329

    Article  CAS  Google Scholar 

  • Southworth D, Cresti M (1997) Comparison of flagellated and nonflagellated sperm in plants. Am J Bot 84:1301–1311

    Article  Google Scholar 

  • Theunis CH, Pierson ES, Cresti M (1991) Isolation of male and female gametes in higher plants. Sex Plant Reprod 4:145–154

    Article  Google Scholar 

  • Theunis CH, Pierson ES, Cresti M (1992) The microtubule cytoskeleton and the rounding of isolated generative cells of Nicotiana tabacum. Sex Plant Reprod 5:64–71

    Article  Google Scholar 

  • Tian HQ, Zhang Z, Russell SD (2001) Sperm dimorphism in Nicotiana tabacum L. Sex Plant Reprod 14:123–125

    Article  Google Scholar 

  • Tian HQ, Yuan T, Russell SD (2005) Relationship between double fertilization and the cell cycle in male and female gametes of tobacco. Sex Plant Reprod 17:243–252

    Article  Google Scholar 

  • Twell D, Park SK, Hawkins TJ, Schubert D, Schmidt R, Smertenko A, Hussey PJ (2002) MOR1/GEM1 has an essential role in the plant-specific cytokinetic phragmoplast. Nat Cell Biol 4:711–714

    Article  CAS  PubMed  Google Scholar 

  • Weterings K, Russell SD (2004) Experimental analysis of the fertilization process. Plant Cell 16:107–118

    Article  Google Scholar 

  • Yang YH, Qiu YL, Xie CT, Tian HQ, Zhang Z, Russell SD (2005) Isolation of two populations of sperm cells and microelectrophoresis of pairs of sperm cells from pollen tubes of tobacco (Nicotiana tabacum). Sex Plant Reprod 18:47–53

    Article  Google Scholar 

  • Zhang Z, Russell SD (1999) Sperm cell surface characteristics of Plumbago zeylanica L. in relation to transport in the embryo sac. Planta 208:539–544

    Article  CAS  Google Scholar 

  • Zhang Z, Xu H, Singh MB, Russell SD (1998) Isolation and collection of two population of viable sperm cells from the pollen of Plumbago zeylanica. Zygote 6:295–298

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to the Nikon Imaging Center at Hokkaido University for assistance with confocal microscopy, image acquisition, and analysis. This work was partly supported by a Grant-in-Aid for Young Scientists from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and by a grant from the Takeda Science Foundation.

Author information

Authors and Affiliations

  1. Division of Innovative Research, Creative Research Initiative ‘Sousei’ (CRIS), Hokkaido University, Kita 21, Nishi 10, Kita-ku, Sapporo, 001-0021, Japan

    Tomonari Hirano & Yoichiro Hoshino

  2. Field Science Center for Northern Biosphere, Hokkaido University, Kita 11, Nishi 10, Kita-ku, Sapporo, 060-0811, Japan

    Yoichiro Hoshino

Authors
  1. Tomonari Hirano
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Yoichiro Hoshino
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence to Yoichiro Hoshino.

Additional information

Communicated by Scott Russell.

Electronic supplementary material

Below is the link to the electronic supplementary material.

497_2009_123_MOESM1_ESM.tif

Fig. S1 DAPI staining of nuclei in a pollen tube cultured for 6 h. The pollen tube was observed under bright field (a) and fluorescence (b) followed by DAPI staining. Arrowheads and arrows indicate vegetative nucleus and generative cell, respectively. Bar = 100 μm (TIFF 1.30 mb)

Supplementary material 2 (DOC 39 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hirano, T., Hoshino, Y. Sperm dimorphism in terms of nuclear shape and microtubule accumulation in Cyrtanthus mackenii . Sex Plant Reprod 23, 153–162 (2010). https://doi.org/10.1007/s00497-009-0123-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00497-009-0123-2

Keywords