Abstract
Pollen tubes grow rapidly in a strictly polarized manner as they transport male reproductive cells through female flower tissues to bring about fertilization. Vegetative pollen tube cells are an excellent model system to investigate processes underlying directional cell expansion. In this chapter, we describe materials and methods required for (1) the identification of novel factors essential for polarized cell growth through the isolation and analysis of Arabidopsis mutants with defects in pollen tube growth and (2) the detailed functional characterization of pollen tube proteins based on transient transformation and microscopic analysis of cultured tobacco pollen tubes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bedinger, P. A., Hardeman, K. J., and Loukides, C. A. (1994) Travelling in style: The cell biology of pollen. Trends Cell Biol 4, 132–138.
Johnson, M. A., von Besser, K., Zhou, Q., Smith, E., Aux, G., Patton, D., Levin, J. Z., and Preuss, D. (2004) Arabidopsis hapless mutations define essential gametophytic functions. Genetics 168, 971–982.
Klahre, U., Becker, C., Schmitt, A. C., and Kost, B. (2006) Nt-RhoGDI2 regulates Rac/Rop signaling and polar cell growth in tobacco pollen tubes. Plant J 46, 1018–1031.
Kost, B., Spielhofer, P., and Chua, N.-H. (1998) A GFP-mouse talin fusion protein labels plant actin filaments in vivo and visualizes the actin cytoskeleton in growing pollen tubes. Plant J 16, 393–401.
Gu, Y., Fu, Y., Dowd, P., Li, S., Vernoud, V., Gilroy, S., and Yang, Z. (2005) A Rho family GTPase controls actin dynamics and tip growth via two counteracting downstream pathways in pollen tubes. J Cell Biol 169, 127–138.
Helling, D., Possart, A., Cottier, S., Klahre, U., and Kost, B. (2006) Pollen tube tip growth depends on plasma membrane polarization mediated by Tobacco PLC3 activity and endocytic membrane recycling. Plant Cell 18, 3519–3534.
Boavida, L. C. and McCormick, S. (2007) Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana. Plant J 52, 570–582.
Read, S. M., Clarke, A. E., and Bacic, A. (1993) Stimulation of growth of cultured Nicotiana tabacum W38 pollen tubes by poly(ethylene glycol) and Cu(II) salts. Protoplasma 177, 1–14.
Read, S. M., Clarke, A. E., and Bacic, A. (1993) Requirements for division of the generative nucleus in cultured pollen tubes of Nicotiana. Protoplasma 174, 101–115.
Smyth, D. R., Bowman, J. L., and Meyerowitz, E. M. (1990) Early flower development in Arabidopsis. Plant Cell 2, 755–767.
Hicks, G. R., Rojo, E., Hong, S., Carter, D. G., and Raikhel, N. V. (2004) Geminating pollen has tubular vacuoles, displays highly dynamic vacuole biogenesis, and requires VACUOLESS1 for proper function. Plant Physiol 134, 1227–1239.
Palanivelu, R. and Preuss, D. (2006) Distinct short-range ovule signals attract or repel Arabidopsis thaliana pollen tubes in vitro. BMC Plant Biol 6, 7.
Faure, J. E., Rotman, N., Fortune, P., and Dumas, C. (2002) Fertilization in Arabidopsis thaliana wild type: Developmental stages and time course. Plant J 30, 481–488.
Sessions, A., Burke, E., Presting, G., Aux, G., McElver, J., Patton, D., Dietrich, B., Ho, P., Bacwaden, J., Ko, C., Clarke, J. D., Cotton, D., Bullis, D., Snell, J., Miguel, T., Hutchison, D., Kimmerly, B., Mitzel, T., Katagiri, F., Glazebrook, J., Law, M., and Goff, S. A. (2002) A high-throughput Arabidopsis reverse genetics system. Plant Cell 14, 2985–2994.
Alonso, J. M., Stepanova, A. N., Leisse, T. J., Kim, C. J., Chen, H., Shinn, P., Stevenson, D. K., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, C., Heller, C., Jeske, A., Koesema, E., Meyers, C. C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D., Carter, D. E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C. C., and Ecker, J. R. (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301, 653–657.
Preuss, D., Rhee, S. Y., and Davis, R. W. (1994) Tetrad analysis possible in Arabidopsis with mutation of the QUARTET (QRT) genes. Science 264, 1458–1460.
Johnson-Brousseau, S. A. and McCormick, S. (2004) A compendium of methods useful for characterizing Arabidopsis pollen mutants and gametophytically-expressed genes. Plant J 39, 761–775.
von Besser, K., Frank, A. C., Johnson, M. A., and Preuss, D. (2006) Arabidopsis HAP2 (GCS1) is a sperm-specific gene required for pollen tube guidance and fertilization. Development 133, 4761–4769.
Twell, D., Yamaguchi, J., Wing, R. A., Ushiba, J., and McCormick, S. (1991) Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev 5, 496–507.
Acknowledgments
B.K. was supported by DFG, BBSRC, FORMAS, and VR funding. M.J. was supported by NSF grant IOS-0644623 and would like to thank Rebecca Macri for the illustration in Fig. 2 and Alexander R. Leydon for micrographs.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Johnson, M.A., Kost, B. (2010). Pollen Tube Development. In: Hennig, L., Köhler, C. (eds) Plant Developmental Biology. Methods in Molecular Biology, vol 655. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-765-5_11
Download citation
DOI: https://doi.org/10.1007/978-1-60761-765-5_11
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-764-8
Online ISBN: 978-1-60761-765-5
eBook Packages: Springer Protocols