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Heat stress affects the cytoskeleton and the delivery of sucrose synthase in tobacco pollen tubes

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Abstract

Main conclusion

Heat stress changes isoform content and distribution of cytoskeletal subunits in pollen tubes affecting accumulation of secretory vesicles and distribution of sucrose synthase, an enzyme involved in cell wall synthesis.

Plants are sessile organisms and are therefore exposed to damages caused by the predictable increase in temperature. We have analyzed the effects of temperatures on the development of pollen tubes by focusing on the cytoskeleton and related processes, such as vesicular transport and cell wall synthesis. First, we show that heat stress affects pollen germination and, to a lesser extent, pollen tube growth. Both, microtubules and actin filaments, are damaged by heat treatment and changes of actin and tubulin isoforms were observed in both cases. Damages to actin filaments mainly concern the actin array present in the subapex, a region critical for determining organelle and vesicle content in the pollen tube apex. In support of this, green fluorescent protein-labeled vesicles are arranged differently between heat-stressed and control samples. In addition, newly secreted cell wall material (labeled by propidium iodide) shows an altered distribution. Damage induced by heat stress also extends to proteins that bind actin and participate in cell wall synthesis, such as sucrose synthase. Ultimately, heat stress affects the cytoskeleton thereby causing alterations in the process of vesicular transport and cell wall deposition.

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Abbreviations

HSP:

Heat-shock protein

PI:

Propidium iodide

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Acknowledgments

We thank the employees of the Botanical Garden of University of Siena for kindly supporting us and for growing tobacco plants.

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  1. Dipartimento Scienze della Vita, Università di Siena, via Mattioli 4, 53100, Siena, Italy

    Luigi Parrotta, Claudia Faleri, Mauro Cresti & Giampiero Cai

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  1. Luigi Parrotta
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  2. Claudia Faleri
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  3. Mauro Cresti
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  4. Giampiero Cai
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Correspondence to Giampiero Cai.

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Online Resource 1a, b, c

Original images of actin filaments in control pollen tubes without the overlay used by ImageJ to calculate the anisotropy of cytoskeleton. Arrows in c show the presence of the actin fringe. Bars 10 µm. (PDF 543 kb)

Online Resource 2

Control of immunogold labeling by omitting primary antibody to sucrose synthase. No signal was detected. Bar 500 nm. 2 (PDF 646 kb)

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Parrotta, L., Faleri, C., Cresti, M. et al. Heat stress affects the cytoskeleton and the delivery of sucrose synthase in tobacco pollen tubes. Planta 243, 43–63 (2016). https://doi.org/10.1007/s00425-015-2394-1

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