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Utilization of Different Bmy1 Intron III Alleles for Predicting β-Amylase Activity and Thermostability in Wild and Cultivated Barley

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Abstract

The third intron of barley (Hordeum vulgare L.) β-amylase 1 (Bmy1) is extremely polymorphic. The use of specific insertion/deletions (indels) in the third intron as markers for cultivar development has been recommended based on associations with β-amylase activity and thermostability. The third intron of Bmy1 in 40 barley genotypes was sequenced and aligned with 15 Bmy1 intron III sequences from GenBank and four alleles (Bmy1.a, Bmy1.b, Bmy1.c, and Bmy1.d) were identified based on indels of 126, 38, 11, and 21 bp. β-Amylase activity and thermostability were assayed in 22 North American cultivars and 12 wild barley genotypes. Cultivars carrying the Bmy1.a and Bmy1.b alleles had β-amylase activity ranges calculated on a fresh weight (FW) basis of 1.8- and 1.5-fold, respectively, and thermostability ranges of 8.8- and 1.2-fold, respectively. β-Amylase activity calculated on a protein basis yielded a 2.4- and 1.4-fold range for Bmy1.a and Bmy1.b, respectively. Significantly different activities were observed in cultivars carrying either Bmy1.a or the Bmy1.b allele when calculated on a FW basis and the Bmy1.a allele when calculated on a protein basis. Significantly different thermostabilities were observed in cultivars carrying the Bmy1.a allele. Wild barleys were found to carry Bmy1.a, Bmy1.b, and Bmy1.c alleles with β-amylase activity ranges calculated on a FW basis of 1.7-, 1.7-, and 2.6-fold, respectively, and thermostability ranges of 1.3-, 1.4-, and 2.1-fold, respectively. β-Amylase activity measured on a protein basis identified a 1.3-, 1.4-, and 2.1-fold range for Bmy1.a, Bmy1.b, and Bmy1.c, respectively. Significantly different activities were found in genotypes with any of these three alleles when calculated on a FW basis yet only in those with the Bmy1.c allele when calculated on a protein basis. Significantly different thermostabilities in genotypes carrying either the Bmy1.b or Bmy1.c allele were observed. In the germplasm studied here, the Bmy1 intron III alleles are not reliable predictors of β-amylase activity and thermostability.

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Acknowledgments

This research was supported by a USDA–ARS, USDA–CREES US Barley Genome Project Special Grant. We thank Charles Karpelenia, Joel Phillips, Mariah Peronto, and Lauren Boerboom for their excellent technical assistance and Brian Steffenson and Mike Casler for their insightful and useful comments. We thank Blake Cooper, Brian Steffenson, Patrick Hayes, and Harold Bockelman for their generous gifts of germplasm that made this study possible.

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Authors and Affiliations

  1. Department of Agronomy, University of Wisconsin-Madison, 1575 Linden Drive, Madison, WI, 53706, USA

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

  2. USDA–ARS Cereal Crops Research Unit, 502 Walnut Street, Madison, WI, 53726, USA

    Cynthia A. Henson

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  1. Marcus A. Vinje
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  2. Stanley H. Duke
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  3. Cynthia A. Henson
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Correspondence to Cynthia A. Henson.

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Vinje, M.A., Duke, S.H. & Henson, C.A. Utilization of Different Bmy1 Intron III Alleles for Predicting β-Amylase Activity and Thermostability in Wild and Cultivated Barley. Plant Mol Biol Rep 28, 491–501 (2010). https://doi.org/10.1007/s11105-009-0168-2

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