Abstract
Given the important role that starch plays in food and non-food uses of many crops, particularly wheat, efforts are being made to manipulate its composition through modification of the amylose/amylopectin ratio. Approaches used to achieve this goal include the manipulation of the genes involved in the starch biosynthetic pathway using natural or induced mutations and transgenic methods. The use of mutagenesis to produce novel allelic variation represents a powerful tool to increase genetic diversity and this approach seems particularly appropriate for starch synthase genes for which limited variation exists. In this work, an EMS-mutagenised population of bread wheat cv. Cadenza has been screened by combining SDS–PAGE analysis of granule bound starch proteins with a TILLING (Targeting Induced Local Lesions IN Genomes) approach at the gene level. In particular we have focused on two groups of synthase genes, those encoding the starch synthase II (Sgp-1) and those corresponding to the waxy proteins (Wx). SDS–PAGE analysis of granule bound proteins allowed the identification of single null genotypes associated with each of the three homoeologous loci. Molecular characterization of induced mutants has been performed using genome specific primer pairs for Sgp-1 and Wx genes. Additional novel allelic variation has also been detected at the different Sgp-1 homoeoloci by using a reverse genetic approach (TILLING). In particular single nucleotide substitutions, introducing a premature stop codon and creating amino acid substitutions, have been identified.
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Acknowledgments
This study is partially financially supported by the European Commission in the Communities 6th Framework Programme, Project HEALTHGRAIN (Food-CT-2005-514008). It reflects the authors’ views and the Community is not liable for any use that may be made of the information contained in this publication. Construction of the mutagenized population of wheat cv. Cadenza and establishment of the TILLING platform was carried out within the Wheat Genetic Improvement Network funded by the Department for Environment, Food and Rural Affairs of the UK.
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Sestili, F., Botticella, E., Bedo, Z. et al. Production of novel allelic variation for genes involved in starch biosynthesis through mutagenesis. Mol Breeding 25, 145–154 (2010). https://doi.org/10.1007/s11032-009-9314-7
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DOI: https://doi.org/10.1007/s11032-009-9314-7