Isolation of differentially expressed genes in wheat caryopses with contrasting starch granule size

Abstract

In order to identify genes responsible for starch granule initiation during early development of wheat caryopsis, nine winter wheat breeding lines were studied. Two breeding lines, which are the most diverse in A-type granule size (26.85 µm versus 23.65 µm) were chosen for further differential gene expression analysis in developing caryopses at 10 and 15 days post-anthesis (DPA). cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis resulted in 384 transcript-derived fragments, out of which 18 were identified as being differentially expressed. Six differentially expressed genes, together with the six well-known starch biosynthesis genes, were chosen for semi-quantitative gene expression analysis in developing wheat caryopses at 10 and 15 DPA. This study provides genomic information on 18 genes differentially expressed at early stages of wheat caryopses development and reports on the identification of genes putatively involved in the production of large A-type granules. These genes are targets for further validation on their role in starch granule synthesis control and provide the basis for the development of DNA marker tools in winter wheat breeding for enhanced starch quality.

Nonsense-mediated decay of sucrose synthase 1 mRNA with induced premature chain termination codon during cold acclimation in winter wheat

“…The cDNA-AFLP procedure was conducted as described earlier (Armonienė et al, 2013a) with minor modifications. Poly(A)+RNA was isolated from the total RNA with a Dynabeads mRNA Purification Kit (Invitrogen Dynal AS, Oslo, Norway).…”