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Differential expression of two β-amylase genes (Bmy1 and Bmy2) in developing and mature barley grain

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Abstract

Two barley (Hordeum vulgare L.) β-amylase genes (Bmy1 and Bmy2) were studied during the late maturation phase of grain development in four genotypes. The Bmy1 and Bmy2 DNA and amino acid sequences are extremely similar. The largest sequence differences are in the introns, seventh exon, and 3′ UTR. Accumulation of Bmy2 mRNA was examined in developing grain at 17, 19, and 21 days after anthesis (DAA). One genotype, PI 296897, had significantly higher Bmy2 RNA transcript accumulation than the other three genotypes at all developmental stages. All four genotypes had Bmy2 mRNA levels decrease from 17 to 19 DAA, and remain the same from 19 to 21 DAA. Levels of Bmy1 mRNA were twenty thousand to over one hundred thousand times more than Bmy2 mRNA levels in genotypes Legacy, Harrington, and Ashqelon at all developmental stages and PI 296897 at 19 and 21 DAA. PI 296897 had five thousand times more Bmy1 mRNA than Bmy2 mRNA at 17 DAA. However, Bmy2 protein was not found at 17 DAA in any genotype. The presence of Bmy2 was immunologically detected at 19 DAA and was present in greater amounts at 21 DAA. Also, Bmy2 protein was found to be stored in mature grain and localized in the soluble fraction. However, Bmy1 protein was far more prevalent than Bmy2 at all developmental stages in all genotypes. Thus, the vast majority of β-amylase activity in developing and mature grain can be attributed to endosperm-specific β-amylase.

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Abbreviations

DAA:

Days after anthesis

RT-qPCR:

Reverse transcription-quantitative polymerase chain reaction

SNP:

Single nucleotide polymorphism

RER:

Relative expression ratio

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Acknowledgments

We thank Charles Karpelenia for his excellent technical assistance and Dr. Ron Skadsen for the generous donation of antibodies. Research supported by USDA-ARS and USDA-CREES U.S. Barley Genome Project Special Grant. Mention of a proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other suitable products.

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Authors and Affiliations

  1. Department of Agronomy, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Marcus A. Vinje, Stanley H. Duke & Cynthia A. Henson

  2. Department of Plant Pathology, University of Wisconsin-Madison, Madison, WI, 53706, USA

    David K. Willis

  3. United States Department of Agriculture-Agricultural Research Service, Cereal Crops Research Unit, Madison, WI, 53726, USA

    Marcus A. Vinje & Cynthia A. Henson

  4. United States Department of Agriculture-Agricultural Research Service, Vegetable Crops Research Unit, Madison, WI, 53706, USA

    David K. Willis

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  1. Marcus A. Vinje
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  2. David K. Willis
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  4. Cynthia A. Henson
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Correspondence to Cynthia A. Henson.

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Vinje, M.A., Willis, D.K., Duke, S.H. et al. Differential expression of two β-amylase genes (Bmy1 and Bmy2) in developing and mature barley grain. Planta 233, 1001–1010 (2011). https://doi.org/10.1007/s00425-011-1348-5

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  • DOI: https://doi.org/10.1007/s00425-011-1348-5

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