Title:
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Charcterising mismatch repair in wheat (triticum aestivum)
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To meet the nutritional requirements of a growing population, world agricultural output
must increase by 70-100% by 2050. However, worldwide wheat yields are close to reaching
a 'glass ceiling'. In order to meet the challenges of a rapidly growing population, scarcity
of resources and a trend toward high-protein diets, traditional agriculture has looked
towards molecular biology for insight and innovation for the future. Studying the
mechanisms underlying DNA damage and repair is crucially important to facilitating
generation of novel variation and developing breeding techniques.
The Mismatch Repair (MMR) system is a highly-conserved series of pathways responsible
for maintaining genomic integrity in prokaryotes and eukaryotes. The constituent proteins
repair DNA mismatches and prevent meiotic recombination between homoeologous
sequences. Work in Arabidopsis, tobacco and tomato has shown that plants with an
impaired MMR system accumulate point mutations and undergo increased
homoeologous recombination, which may be useful in breeding programs.
Two key MMR genes are explored: MutS Homologue 2 (MSH2) acts as a heterodimer with
MSH3, MSH6 or MSH7 to detect point mutations, insertions and deletions. Post Meiotic
Segregation increased 2 (PMS2) is a homologue of the MutL gene, which co-ordinates repair
of mismatches. The two genes were cloned and sequenced from elite cultivar Cadenza, and
the sequences subsequently compared to next generation sequencing data from Chinese
Spring. Three homoeologous copies of each gene were identified in the genome of wheat,
and two of these were sequenced from the transcriptome.
The expression of MSH2 was studied using qPCR in the transcriptome of Cadenza and four
gamma-irradiated deletion lines, generated in a Paragon background. The four deletion
lines showed varying responses: from down-regulation of MSH2 expression by 57% to upregulation
by 82%. The expression of the individual homoeologues of MSH2 were also
explored using a qPCR based method and by examining RNAseq data, but this was
inconclusive.
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