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Laser capture microdissection and cDNA microarrays used to generate gene expression profiles of the rapidly expanding fibre initial cells on the surface of cotton ovules

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Abstract

Cotton (Gossypium hirsutum L.) fibre initial cells undergo a rapid cellular re-programming around anthesis to form the long cellulose fibres prized for textile manufacture. On the day of anthesis the cotton fibre initial cells balloon out from the ovule surface and so are clearly distinguished from adjacent epidermal pavement cells. To enhance our understanding of the molecular processes that determine which cells become fibres and why adjacent epidermal cells remain in a different developmental state we studied the expression profiles of the two respective cell types. Using laser-capture microdissection, coupled with an in vitro RNA amplification system, we used cDNA microarray slides to profile the gene expression in expanding fibre initials compared to the non-expanding epidermal cells at an early stage just after the fibre initials are discernable. Except for a few regulatory genes, the genes that are up-regulated in the cotton fibre initials relative to epidermal cells predominantly encode proteins involved in generating the components for the extra cell membrane and primary cell wall needed for the rapid cell expansion of the initials. This includes synthesis of enzymes and cell wall proteins, carbohydrates, and lipids. An analysis of single channel fluorescence levels confirmed that these classes of genes were also the most highly expressed genes in fibre initials. Genes involved in DNA metabolism were also well represented in the expanding fibre cell, consistent with the limited endoreduplication we previously reported to occur in fibre initial cells.

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Abbreviations

ACC:

1-Aminocyclopropane-1-carboxylic acid

aRNA:

Amplified RNA

DP16:

Cultivar Deltapine 16

dpa:

Days post anthesis

GEO:

Gene expression omnibus

GO:

Gene ontology

LCM:

Laser-capture microdissection

SAM:

S-adenosyl-l-methionine

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Acknowledgements

This work was partly funded by the Australian Cotton Research and Development Corporation and Cotton Seed Distributors, Wee Waa, NSW. We thank Todd Collins for his excellent technical assistance, the University of Western Australia for access to their laser capture microscope, and Gavin Kennedy for curation of the microarray data in the CSIRO Gena database. Iain Wilson, Glenn Stone, David Lovell, Rob Dunne, Peter Baker, and Maree O’Sullivan (CSIRO) provided valuable discussions and advice on the microarray analyses.

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Author notes

  1. Yingru Wu

    Present address: Food Standards Australia New Zealand, P.O. Box 7186, Canberra BC, ACT, 2610, Australia

Authors and Affiliations

  1. CSIRO Plant Industry, P.O. Box 1600, Canberra, ACT, 2601, Australia

    Yingru Wu, Danny J. Llewellyn, Rosemary White, Yves Al-Ghazi & Elizabeth S. Dennis

  2. CSIRO Mathematical and Information Sciences, GPO Box 664, Canberra, ACT, 2601, Australia

    Katya Ruggiero

  3. NSW Agricultural Genomics Centre, P.O. Box 1600, Canberra, ACT, 2601, Australia

    Yingru Wu, Katya Ruggiero & Elizabeth S. Dennis

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  1. Yingru Wu
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  2. Danny J. Llewellyn
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Correspondence to Danny J. Llewellyn.

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Wu, Y., Llewellyn, D.J., White, R. et al. Laser capture microdissection and cDNA microarrays used to generate gene expression profiles of the rapidly expanding fibre initial cells on the surface of cotton ovules. Planta 226, 1475–1490 (2007). https://doi.org/10.1007/s00425-007-0580-5

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