Journal article
Genotyping single nucleotide polymorphisms for selection of barley β-amylase alleles
Plant Molecular Biology Reporter, Vol.20(2), pp.149-159
2002
Abstract
A high-throughput single nucleotide polymorphism (SNP) genotyping system was developed and used to select barley seedlings carrying superior alleles of β-amylase. In the malting process, β-amylase is a key enzyme involved in the degradation of starch. Four allelic forms of the enzyme are found in barley, each exhibiting a different rate of thermal inactivation, or thermostability. The level of thermostability influences starch degradation, which determines the yield of fermentable sugars for alcohol production during brewing. Control of the fermentability level is important for barley breeding programs to allow targeting quality profiles of new varieties to suit end-user requirements. Alignment of the cDNA sequences encoding the 4 enzyme forms revealed 6 SNPs (cSNPs). The 4 alleles could be identified unambiguously by codominantly genotyping 2 of the cSNPs using a duplex single nucleotide primer extension (SNuPE) assay. Two genotyping primers with their 3' ends directly flanking the selected SNPs were annealed to the amplified target sequences and extended by single dideoxynucleotides complementary to the polymorphic nucleotides. Extended primers were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS). Making use of the inherent molecular weight difference between DNA bases, incorporated nucleotides were identified by the increase in mass of the extended primers. A cleaved amplified polymorphic sequence (CAPS) assay enabling broader classification of the alleles was also developed to facilitate the transfer of this marker to other laboratories. Plants carrying alternative β-amylase alleles were selected at the seedling stage for barley breeding.
Details
- Title
- Genotyping single nucleotide polymorphisms for selection of barley β-amylase alleles
- Authors/Creators
- M. Paris (Author/Creator) - Murdoch UniversityM.G.K. Jones (Author/Creator) - Murdoch UniversityJ.K. Eglinton (Author/Creator) - The University of Adelaide
- Publication Details
- Plant Molecular Biology Reporter, Vol.20(2), pp.149-159
- Publisher
- Springer New York
- Identifiers
- 991005541348007891
- Copyright
- © International Society for Plant Molecular Biology 2002
- Murdoch Affiliation
- State Agricultural Biotechnology Centre
- Language
- English
- Resource Type
- Journal article
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Source: InCites
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- Collaboration types
- Domestic collaboration
- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.85 Food Science & Technology
- 3.85.99 Cereal Starch Properties
- Web Of Science research areas
- Biochemical Research Methods
- Plant Sciences
- ESI research areas
- Plant & Animal Science