Abstract
The question of how pollen tubes orient themselves on their way to the egg cell is a major focus of plant reproduction research. The role of physical guidance through the tissues of the pistil in relation to the mechanical perception and growth adaptation of the pollen tubes has not been sufficiently investigated. In order to advance research on the mechanical perception of pollen tubes and their force application during invasive growth, we present simple methods for the observation and mechanical characterization of pollen tubes in vitro, which can be established with little effort in any biological laboratory with standard equipment. Pollen grains are germinated in a hydrogel containing agarose and their growth is recorded in 3D using brightfield microscopy. Using suitable analysis software, parameters such as growth rate and pollen tube diameter can then be determined to estimate the exerted penetration force.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Palanivelu R, Tsukamoto T (2012) Pathfinding in angiosperm reproduction: pollen tube guidance by pistils ensures successful double fertilization. Wiley Interdiscip Rev Dev Biol 1:96–113. https://doi.org/10.1002/wdev.6
Sanati Nezhad A, Naghavi M, Packirisamy M et al (2013) Quantification of cellular penetrative forces using lab-on-a-chip technology and finite element modeling. Proc Natl Acad Sci 110:8093. https://doi.org/10.1073/pnas.1221677110
Burri JT, Vogler H, Läubli NF et al (2018) Feeling the force: how pollen tubes deal with obstacles. New Phytol 220:187–195. https://doi.org/10.1111/nph.15260
Gossot O, Geitmann A (2007) Pollen tube growth: coping with mechanical obstacles involves the cytoskeleton. Planta 226:405–416. https://doi.org/10.1007/s00425-007-0491-5
Ghanbari M, Packirisamy M, Geitmann A (2018) Measuring the growth force of invasive plant cells using Flexure integrated Lab-on-a-Chip (FiLoC). Technology 06:101–109. https://doi.org/10.1142/S2339547818500061
Cheung AY, Wu H-M, di Stilio V et al (2000) Pollen-Pistil interactions in Nicotiana tabacum. Ann Bot 85:29–37. https://doi.org/10.1006/anbo.1999.1016
Lennon KA, Roy S, Hepler PK, Lord EM (1998) The structure of the transmitting tissue of Arabidopsis thaliana (L.) and the path of pollen tube growth. Sex Plant Reprod 11:49–59. https://doi.org/10.1007/s004970050120
Brewbaker JL, Kwack BH (1963) The essential role of calcium in pollen germination and pollen tube growth. Am J Bot 50:859–865. https://doi.org/10.1002/j.1537-2197.1963.tb06564.x
Gerum RC, Richter S, Fabry B, Zitterbart DP (2017) ClickPoints: an expandable toolbox for scientific image annotation and analysis. Methods Ecol Evol 8:750–756. https://doi.org/10.1111/2041-210X.12702
Fakhouri S, Hutchens SB, Crosby AJ (2015) Puncture mechanics of soft solids. Soft Matter 11:4723–4730. https://doi.org/10.1039/C5SM00230C
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Reimann, R., Kah, D. (2020). Characterization of Growth Behavior and the Resulting Forces Applied by Pollen Tubes in a 3D Matrix. In: Geitmann, A. (eds) Pollen and Pollen Tube Biology. Methods in Molecular Biology, vol 2160. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0672-8_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0672-8_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0671-1
Online ISBN: 978-1-0716-0672-8
eBook Packages: Springer Protocols