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Characterisation and functional analysis of two barley caleosins expressed during barley caryopsis development

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Abstract

Two full-length cDNA sequences homologous to caleosin, a seed-storage oil-body protein from sesame, were identified from a series of barley grain development cDNA libraries and further characterised. The cDNAs, subsequently termed HvClo1 and HvClo2, encode proteins of 34 kDa and 28 kDa, respectively. Real-time RT-PCR indicated that HvClo1 is expressed abundantly during the later stages of embryogenesis and is seed-specific, accumulating in the scutellum of mature embryos. HvClo2 is expressed mainly in the endosperm tissues of the developing grain. We show that HvClo1 and HvClo2 are paralogs that co-segregate on barley chromosome 2HL. Transient expression of HvClo1 in lipid storage and non-storage cells of barley using biolistic particle bombardment indicates that caleosins have different subcellular locations from the structural oil-body protein oleosin, and by inference participate in different sorting pathways. We observe that caleosin sorts via small vesicles, suggesting a likely association with lipid trafficking, membrane expansion and oil-body biogenesis.

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Abbreviations

EST:

Expressed sequence tag

DPA:

Days post-anthesis

GFP:

Green fluorescent protein

RFP:

Red fluorescent protein

SNP:

Single-nucleotide polymorphism

TAG:

Triacylglyceride

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Acknowledgements

This work was supported by the Scottish Executive Environment and Rural Affairs Department. We thank Professor Jason Tzen for kindly providing the anti-sesame caleosin sera, Ms Clare Booth for DNA sequencing, Ms Diane Davidson for demonstrating the transient expression experiment, Dr Christophe Lacomme for suggestions and ideas, Dr Gordon Jamieson for analysing caleosin sequences and Dr Luke Ramsay for genetic mapping.

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Authors and Affiliations

  1. Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, Scotland, UK

    Hui Liu, Peter Hedley, Linda Cardle, Kathryn M. Wright, Ingo Hein, David Marshall & Robbie Waugh

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  1. Hui Liu
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  2. Peter Hedley
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  3. Linda Cardle
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  4. Kathryn M. Wright
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  5. Ingo Hein
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  7. Robbie Waugh
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Correspondence to Robbie Waugh.

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Liu, H., Hedley, P., Cardle, L. et al. Characterisation and functional analysis of two barley caleosins expressed during barley caryopsis development. Planta 221, 513–522 (2005). https://doi.org/10.1007/s00425-004-1465-5

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