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Storage-protein regulation and lipid accumulation in microspore embryos of Brassica napus L.

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Abstract

Embryos derived in vitro from isolated microspores of Brassica napus L. were compared with their zygotic counterparts. Parameters investigated included storage-protein accumulation and gene expression, fattyacid composition, storage-lipid biosynthesis, and the appearance of oil-body proteins. The microspore embryos accumulate storage-protein and show increases in levels of their transcripts during the torpedo stage. These embryos were sensitive to abscisic acid (ABA) with respect to accumulation of storage-protein mRNA and oil-body proteins. Post-transcriptional regulation of cruciferin accumulation is indicated by a disparity between ABA-enhanced transcript accumulation and a less marked effect at the level of protein accumulation. To investigate storage-lipid profiles, two cultivars of Brassica napus, Reston and Topas, were used. The former accumulates major quantities of C20 (11.2%) and C22 (39.9%) fatty acids in its seeds, the latter predominantly C18 fatty acids. The higher-molecular-weight fatty acids (>C18) normally occur only in seeds and were used as biochemical markers for seed-specific metabolism in microspore embryos. Microspore embryos from Reston were found to accumulate C20 (10.6%) and C22 (31.2%) fatty acids after 35 d in culture at levels and proportions comparable to those found in seeds. Similarly, microspore embryos of Topas had a fatty-acid profile similar to that of mature Topas seed. Activities of enzymes involved in the accumulation of storage lipids (erucoyl-CoA synthetase [EC 6.2.1.3], erucoyl-CoA thioesterase [EC 3.1.2.2] and erucoyl-CoA acyltransferase [EC 2.3.1.15 or EC 2.3.1.20]) were detected in torpedostage microspore embryos. Their specific activities were higher than have been reported to date for analogous preparations from zygotic embryos of B. napus. The similarities in storage-lipid and protein composition of these embryos to their zygotic counterparts, along with their sensitivity to ABA, indicate that microspore embryos might be exploited to facilitate studies of biochemistry and gene regulation in oilseeds.

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Abbreviations

ABA:

abscisic acid

DW:

dry weight

FW:

fresh weight

kDA:

kilodalton

TAG:

triacylglycerol

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Authors and Affiliations

  1. Plant Biotechnology Institute, National Research Council of Canada, 110 Gymnasium Road, S7N 0W9, Saskatoon, Sask., Canada

    David C. Taylor & Edward W. Underhill

  2. Federal Centre for Lipid Research, Institute for Biochemistry and Technology, H.P. Kaufmann Institut, Piusallee 68, D-4400, Münster, Germany

    Nikolaus Weber

  3. Plant Research Centre, Agriculture Canada, K1A 0C6, Ottawa, Ont.

    M. Keith Pomeroy & Wilfred A. Keller

  4. Department of Biological Sciences, University of Calgary, 2500 University Drive NW, T2N 1N4, Calgary, Alta.

    Ron W. Wilen & Maurice M. Moloney

  5. Biotechnica Canada Inc., 6815 8th Street NE, T2E 7H7, Calgary, Alta., Canada

    William R. Scowcroft & Larry A. Holbrook

Authors
  1. David C. Taylor
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  2. Nikolaus Weber
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  3. Edward W. Underhill
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  4. M. Keith Pomeroy
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  5. Wilfred A. Keller
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  6. William R. Scowcroft
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  7. Ron W. Wilen
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  8. Maurice M. Moloney
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Additional information

National Research Council Canada No. 31224

The authors would like to thank Marjorie Norris (Calgary), Nancy Long and John Grainger (Ottawa) for excellent technical assiatance with the preparation and amintenance of cultures. Helpful discussions with Drs. Randall Weselake, Sam MacKenzie, Lois Edwards and Ed Yeung are greatly appreciated. This work was supported in part by the National Research Council of Canada contribution program and by an operating grant (No. A3490) from Natural Sciences and Engineering Research Council of Canada to M.M.M.

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Taylor, D.C., Weber, N., Underhill, E.W. et al. Storage-protein regulation and lipid accumulation in microspore embryos of Brassica napus L.. Planta 181, 18–26 (1990). https://doi.org/10.1007/BF00202320

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