Fine mapping and targeted SNP survey using rice-wheat gene colinearity in the region of the Bo1 boron toxicity tolerance locus of bread wheat
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Toxicity due to high levels of soil boron (B) represents a significant limitation to cereal production in some regions, and the Bo1 gene provides a major source of B toxicity tolerance in bread wheat (Triticum aestivum L.). A novel approach was used to develop primers to amplify and sequence gene fragments specifically from the Bo1 region of the hexaploid wheat genome. Single-nucleotide polymorphisms (SNPs) identified were then used to generate markers close to Bo1 on the distal end of chromosome 7BL. In the 16 gene fragments totaling 19.6 kb, SNPs were observed between the two cultivars Cranbrook and Halberd at a low frequency (one every 613 bp). Furthermore, SNPs were distributed unevenly, being limited to only two genes. In contrast, RFLP provided a much greater number of genetic markers, with every tested gene identifying polymorphism. Bo1 previously known only as a QTL was located as a discrete Mendelian locus. In total, 28 new RFLP, PCR and SSR markers were added to the existing map. The 1.8 cM Bo1 interval of wheat corresponds to a 227 kb section of rice chromosome 6L encoding 21 predicted proteins with no homology to any known B transporters. The co-dominant PCR marker AWW5L7 co-segregated with Bo1 and was highly predictive of B tolerance status within a set of 94 Australian bread wheat cultivars and breeding lines. The markers and rice colinearity described here represent tools that will assist B tolerance breeding and the positional cloning of Bo1.
KeywordsBread Wheat Double Haploid Double Haploid Line Cleave Amplify Polymorphic Sequence Cleave Amplify Polymorphic Sequence Marker
The authors would like to thank K.T. Lawlor for excellent technical assistance, M. Pallotta for unpublished mapping data of SSRs, J.G. Paull for comments, P. Eckermann for statistical advice, and H. O’Sullivan for critical comments on previous versions of the manuscript. While conducting this research T. Schnurbusch was partly supported by a Research Fellowship, Feodor-Lynen-Program, from the Alexander-von-Humboldt Foundation, Bonn-Bad Godesberg, Germany, and partly by the Australian Centre for Plant Functional Genomics (ACPFG), Adelaide, Australia.
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