High resolution melting analysis for the detection of EMS induced mutations in wheat SBEIIa genes

A - Papers appearing in refereed journals

Botticella, E., Sestili, F., Hernandez-Lopez, A., Phillips, A. L. and Lafiandra, D. 2011. High resolution melting analysis for the detection of EMS induced mutations in wheat SBEIIa genes. BMC Plant Biology. 11, p. 156. https://doi.org/10.1186/1471-2229-11-156

AuthorsBotticella, E., Sestili, F., Hernandez-Lopez, A., Phillips, A. L. and Lafiandra, D.
Abstract

Background
Manipulation of the amylose-amylopectin ratio in cereal starch has been identified as a major target for the production of starches with novel functional properties. In wheat, silencing of starch branching enzyme genes by a transgenic approach reportedly caused an increase of amylose content up to 70% of total starch, exhibiting novel and interesting nutritional characteristics.

In this work, the functionality of starch branching enzyme IIa (SBEIIa) has been targeted in bread wheat by TILLING. An EMS-mutagenised wheat population has been screened using High Resolution Melting of PCR products to identify functional SNPs in the three homoeologous genes encoding the target enzyme in the hexaploid genome.

Results
This analysis resulted in the identification of 56, 14 and 53 new allelic variants respectively for SBEIIa-A, SBEIIa-B and SBEIIa-D. The effects of the mutations on protein structure and functionality were evaluated by a bioinformatic approach. Two putative null alleles containing non-sense or splice site mutations were identified for each of the three homoeologous SBEIIa genes; qRT-PCR analysis showed a significant decrease of their gene expression and resulted in increased amylose content. Pyramiding of different single null homoeologous allowed to isolate double null mutants showing an increase of amylose content up to 21% compared to the control.

Conclusion
TILLING has successfully been used to generate novel alleles for SBEIIa genes known to control amylose content in wheat. Single and double null SBEIIa genotypes have been found to show a significant increase in amylose content.

Year of Publication2011
JournalBMC Plant Biology
Journal citation11, p. 156
Digital Object Identifier (DOI)https://doi.org/10.1186/1471-2229-11-156
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
European Union
Funder project or codeCentre for Crop Genetic Improvement (CGI)
Development of the lightscanner as a generic platform for novel allele discovery
Project HEALTHGRAIN
Publisher's version
Output statusPublished
Publication dates
Online10 Nov 2011
Publication process dates
Accepted10 Nov 2011
Copyright licenseCC BY
PublisherBiomed Central Ltd
ISSN1471-2229

Permalink - https://repository.rothamsted.ac.uk/item/8q9ww/high-resolution-melting-analysis-for-the-detection-of-ems-induced-mutations-in-wheat-sbeiia-genes

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