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Microspore-derived cell suspension cultures of oilseed rape as a system for studying gene expression

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Abstract

Abiotic stress, such as extreme temperature, drought, or excessive salinity, is one of the leading causes of crop loss worldwide. Microspore-derived (MD) cell suspension cultures of Brassica napus L. cv. Jet Neuf have been shown to be a useful system for studying the biochemistry of developing oilseeds. In the present study, we describe the application of MD cell suspension cultures of B. napus as a system for studying gene expression in response to abiotic stress, and demonstrate emybryogenic competence in cultures that have been continuously subcultured for more than 20 years. MD cell suspension cultures of B. napus L. cv Jet Neuf were exposed to low temperature or osmotic stress and the expression profile of known stress responsive genes was evaluated. The gene expression profile of BN115, a known cold-responsive gene in B. napus, was similar to that described for intact cold-acclimated plants. Likewise, two late embryogenesis abundant (Lea) genes were shown to be up-regulated in response to low temperature or osmotic stress. The results demonstrate that B. napus MD cell suspension cultures are a useful system for the investigation of changes in gene expression in plants brought about by abiotic stress.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

ABA:

Abscisic acid

BA:

6-Benzylaminopurine

CRBC:

Chicken red blood cells

CTR:

Control

LT:

Low temperature

MD:

Microspore-derived

NAA:

α-Naphthaleneacetic acid

PEG:

Polyethylene glycol

PI:

Propidium iodide

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Acknowledgements

The authors are grateful to Dr. Saleh Shah for discussions on somatic embryogenesis, and Susan Goruk for obtaining chicken red blood cells and assisting with the flow cytometry analysis. As well, the authors thank Drs. Wilf Keller and David C. Taylor for their critical evaluation of the manuscript. This research was funded by Genome Canada, Genome Prairie, the Alberta Agricultural Research Institute, the Alberta Science and Research Authority, a Natural Sciences and Engineering Research Council of Canada Discovery Grant to RJW and the Canada Research Chairs Program.

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Authors and Affiliations

  1. Department of Agricultural, Food & Nutritional Science, University of Alberta, 4-10 Ag/For Building, Edmonton, AB, Canada, T6G 2P5

    Yongzhong Shi, Timothy B. Warrington, Gordon K. Murdoch, E. Chris Kazala, Crystal L. Snyder & Randall J. Weselake

  2. Faculty of Life Science and Technology, Central South Forestry University, Changsha, Hunan, P.R. China, 410004

    Gangbiao Xu

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  1. Yongzhong Shi
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  2. Gangbiao Xu
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  3. Timothy B. Warrington
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  4. Gordon K. Murdoch
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  7. Randall J. Weselake
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Correspondence to Randall J. Weselake.

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Shi, Y., Xu, G., Warrington, T.B. et al. Microspore-derived cell suspension cultures of oilseed rape as a system for studying gene expression. Plant Cell Tiss Organ Cult 92, 131–139 (2008). https://doi.org/10.1007/s11240-007-9313-5

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  • DOI: https://doi.org/10.1007/s11240-007-9313-5

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