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Protein storage vacuoles of Brassica napus zygotic embryos accumulate a BURP domain protein and perturbation of its production distorts the PSV

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Abstract

BNM2 is a prototypical member of the enigmatic BURP domain protein family whose members contain the signature FX6–7GX10–28PX25–31CX11–12X2SX45–56CHX10CHX25–29CHX2TX15–16PX5CH in the C-terminus. This protein family occurs only in plants, and the cognate genes vary very widely in their expression contexts in vegetative and reproductive tissues. None of the BURP family members has been assigned any biochemical function. BNM2 was originally discovered as a gene expressed in microspore-derived embryos (MDE) of Brassica napus but we found that MDE do not contain the corresponding protein. We show that BNM2 protein production is confined to the seeds and localized to the protein storage vacuoles (PSV) even though the transcript is found in vegetative parts and floral buds as well. In developing seeds, transcript accumulation precedes protein appearance by more than 18 days. RNA accumulation peaks at ~20 days post anthesis (DPA) whereas protein accumulation reaches its maximum at ~40 DPA. Transgenic expression of BNM2 does not abrogate this regulation to yield ectopic protein production or to alter the temporal aspect of BNM2 accumulation. Overexpression of BNM2 led to spatial distortion of storage protein accumulation within PSV and to some morphological alterations of PSVs. However, the overall storage protein content was not altered.

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Acknowledgments

We thank Dr. Dwayne Hegedus for cruciferin and napin antibodies and Dr. Alison Ferrie for microspore embryos, Dr. John Kelly for LC-MS/MS of peptides. PT is grateful for Devin Polichuk for valuable discussions. We thank the PBI DNA Technology Unit for DNA synthesis and sequencing. This work is supported by National Research Council of Canada (NRC) and Natural Science and Engineering Research Council of Canada (NSERC). This is NRCC publication 50148. GS dedicates this paper to the memory of his professor Dr. V. N. Iyer.

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  1. Prapapan Teerawanichpan

    Present address: Department of Food and Bioproduct Sciences, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, S7N 5A8, Canada

  2. Qun Xia

    Present address: School of Agriculture, Research, Extension and Applied Sciences, Alcorn State University, 1000 ASU Drive, Alcorn, MS, 39096, USA

Authors and Affiliations

  1. Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, SK, S7N 0W9, Canada

    Prapapan Teerawanichpan, Qun Xia, Raju Datla & Gopalan Selvaraj

  2. Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, S7N 5B4, Canada

    Sarah J. Caldwell

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Correspondence to Gopalan Selvaraj.

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Teerawanichpan, P., Xia, Q., Caldwell, S.J. et al. Protein storage vacuoles of Brassica napus zygotic embryos accumulate a BURP domain protein and perturbation of its production distorts the PSV. Plant Mol Biol 71, 331 (2009). https://doi.org/10.1007/s11103-009-9541-7

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