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Transient expression and analysis of fluorescent reporter proteins in plant pollen tubes

Nature Protocols volume 6, pages 419–426 (2011)Cite this article

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Abstract

The pollen tube is an excellent single-cell model system for studying cellular processes in plant cell biology. This protocol describes a detailed step-by-step procedure with optimized conditions for introducing various fluorescent reporter proteins into lily, tobacco and Arabidopsis pollen grains by means of biolistics for their transient expression and subsequent analysis in germinating pollen tubes. The whole experiment consists of four major stages: coating gold microcarriers with DNA constructs, preparation of pollen grains, transformation of plasmid DNA into pollen grains by particle delivery system and germination of bombarded pollen grains in optimized germination media to obtain pollen tubes for protein trafficking, protein localization, drug treatment and organelle dynamics analysis. This protocol takes about 4–12 h from pollen preparation to protein detection.

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Figure 1: Transient expression of fluorescent reporter proteins in growing lily pollen tubes.
Figure 2: Transient expression of fluorescent reporter proteins in growing tobacco and Arabidopsis pollen tubes.

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Acknowledgements

This work was supported by grants from the Research Grants Council of Hong Kong (CUHK488707, CUHK465708, CUHK466309 and CUHK466610), Chinese University of Hong Kong (CUHK) Schemes B and C, University Grants Committee/Area of Excellence (to L.J.).

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Authors and Affiliations

  1. School of Life Sciences, Centre for Cell and Developmental Biology, The Chinese University of Hong Kong, Hong Kong, China

    Hao Wang & Liwen Jiang

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  1. Hao Wang
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  2. Liwen Jiang
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Contributions

H.W. executed the experiments and wrote the manuscript. L.J. supervised the study and edited the final version of the manuscript.

Corresponding author

Correspondence to Liwen Jiang.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Video 1

Dynamics of GFP-BP-80 in growing lily pollen tube. (MOV 249 kb)

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Wang, H., Jiang, L. Transient expression and analysis of fluorescent reporter proteins in plant pollen tubes. Nat Protoc 6, 419–426 (2011). https://doi.org/10.1038/nprot.2011.309

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