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α-Amylase gene expression during kernel development in relation to pre-harvest sprouting in wheat and triticale

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Abstract

Triticale, an artificial hybrid of wheat and rye, combines the good yield and nutritional value of wheat with good disease resistance of rye. Despite these advantages, pre-harvest sprouting (PHS) can seriously limit triticale production. Although different field and laboratory tests exist, it remains difficult to breed for PHS tolerance and mechanisms behind PHS are still elusive. As part of an integrated study focussing on the genetic and physiological background of PHS, a multidisciplinary approach was pursued monitoring α-amylase, an enzyme that is involved in PHS, both at transcriptional and post-transcriptional levels during kernel development. Plants of PHS-tolerant and PHS-susceptible varieties of wheat and triticale were grown under controlled conditions from flowering to harvest. At regular time points, kernels were harvested for germination tests, α-amylase analysis and RNA extraction. RNA extracts were used in an RT-qPCR assay to obtain expression profiles of the α-Amy1 and α-Amy2 genes. The α-amylase activity profiles during kernel development corresponded well with results from previous reports. The relative gene expression profiles revealed that α-Amy2 peaked in the beginning of kernel development, while α-Amy1 increased towards harvest maturity. Furthermore, the relative α-Amy1 expression was higher in the PHS-susceptible varieties compared to the PHS-tolerant varieties.

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Abbreviations

C q :

Quantification cycle

DM:

Dry matter

DPA:

Days post anthesis

IEF:

Isoelectric focusing

LMA:

Late maturity α-amylase activity

PHS:

Pre-harvest sprouting

pI:

Isoelectric point

PM:

Physiological maturity

RT-qPCR:

Real-time reverse transcriptase polymerase chain reaction

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Acknowledgments

The research of Sarah De Laethauwer was financed by the research funding of University College Ghent. We are also grateful to the Institute for Agricultural and Fisheries Research—Plant Sciences Unit for their expertise and technical assistance. In addition, we would like to thank Dr. Mark D. Wilkinson for his cooperation on the Phadebas assay and Dr. Kolumbina Mrva for her tips on the IEF assay.

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Authors and Affiliations

  1. Associated Faculty of Applied Bioscience Engineering, University College Ghent and Ghent University, Valentin Vaerwyckweg 1, 9000, Ghent, Belgium

    Sarah De Laethauwer, Ingeborg Stals & Geert Haesaert

  2. Plant Sciences Unit, Institute for Agricultural and Fisheries Research, Caritasstraat 21, 9090, Melle, Belgium

    Jan De Riek

  3. Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Dirk Reheul

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  1. Sarah De Laethauwer
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  2. Jan De Riek
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  3. Ingeborg Stals
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  4. Dirk Reheul
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  5. Geert Haesaert
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Correspondence to Sarah De Laethauwer.

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Communicated by A. Gniazdowska-Piekarska.

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De Laethauwer, S., De Riek, J., Stals, I. et al. α-Amylase gene expression during kernel development in relation to pre-harvest sprouting in wheat and triticale. Acta Physiol Plant 35, 2927–2938 (2013). https://doi.org/10.1007/s11738-013-1323-9

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  • DOI: https://doi.org/10.1007/s11738-013-1323-9

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