Abstract
Speltoid spikes are characterized by pyramidal spike morphology featuring an elongated rachis and tenacious glumes. Speltoids are considered undesirable spike aberrants in wheat breeding leading to increased heterogeneity within a cultivar candidate. As a consequence, the presence of speltoids may result in rejection of a cultivar candidate during official field trials or denial of cultivar certification during seed multiplication. A reliable method is, thus, required to assess the occurrence of speltoids, early on in a wheat breeding program. The domestication gene Q located on the long arm of wheat chromosome 5A is known to suppress the speltoid phenotype in wheat. Here, a quantitative pyrosequencing assay was developed to distinguish between normal wheat plants, which possess two copies of the Q allele, and aberrants, which are either aneuploids lacking the correct number of chromosome 5A copies or plants which carry the primitive q allele. An accurate and reproducible determination of the Q gene copy number was achieved for different wheat genotypes based on homoeologous sequence quantification with two primer combinations at the Q locus. Single plants with one to four copies of the Q allele could be detected by quantitative pyrosequencing which corresponded to the occurrence of speltoid (1 Q allele), normal (2 Q alleles), and compact (more than 2 Q alleles) spikes. Q and q specific alleles could be differentiated at SNP position 2299 of the Q gene. This SNP is assumed to be related to the emergence of free-threshing wheat forms. To our knowledge this is the first report for detection of aneuploids and differentiation of Q alleles in bread wheat using pyrosequencing technology. In future, quantitative pyrosequencing assay can be applied in wheat breeding programs to carry out marker-assisted selection against the presence of speltoid spike aberrants.
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Acknowledgments
We are grateful to Ms. Roswitha Ende for excellent technical assistance, and the German wheat breeding companies KWS Lochow GmbH, Bergen, Germany, and W. von Borries-Eckendorf GmbH & Co. KG, Leopoldshöhe, Germany, and the Federal Plant Variety Office, Hannover, Germany, for providing speltoid bread wheat material. We further thank SaKa Pflanzenzucht GmbH & Co. KG, Hamburg, Germany, Saatzucht Josef Breun GmbH & Co. KG, Herzogenaurach, Germany, and Saatzucht Streng GmbH & Co. KG, Uffenheim, Germany, for conducting field trails. This project was financially supported by the Federal Ministry of Economics and Technology (BMWi, code number KF 2104501 MD8) and administrated by “Arbeitsgemeinschaft industrieller Forschungsvereinigungen e.V.” (AiF) and “Gemeinschaft zur Förderung der privaten deutschen Pflanzenzüchtung e.V.” (GFP).
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Förster, S., Schumann, E., Eberhard Weber, W. et al. Discrimination of alleles and copy numbers at the Q locus in hexaploid wheat using quantitative pyrosequencing. Euphytica 186, 207–218 (2012). https://doi.org/10.1007/s10681-011-0561-4
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DOI: https://doi.org/10.1007/s10681-011-0561-4