Targeted modification of storage protein content resulting in improved amino acid composition of barley grain

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  • Md. Shafiqul Islam Sikdar, Department of Agronomy, Hajee Mohammad Danesh Science and Technology University, Danmark
  • S Bowra, Danmark
  • Daiana Schmidt, Departamento de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Danmark
  • Giuseppe Dionisio
  • Preben Bach Holm, Danmark
  • Eva Vincze
C-hordein in barley and ω-gliadins in wheat are members of the prolamins protein families. Prolamins are the major component of cereal storage proteins and composed of non-essential amino acids (AA) such as proline and glutamine therefore have low nutritional value. Using double stranded RNAi silencing technology directed towards C-hordein we obtained transgenic barley lines with up to 94.7 % reduction in the levels of C-hordein protein relative to the parental line. The composition of the prolamin fraction of the barley parental line cv. Golden Promise was resolved using SDS-PAGE electrophoresis, the protein band were excised and the proteins identified by quadrupole-time-of-flight mass spectrometry. Subsequent SDS-PAGE separation and analysis of the prolamin fraction of the transgenic lines revealed a reduction in the amounts of C-hordeins and increases in the content of other hordein family members. Analysis of the AA composition of the transgenic lines showed that the level of essential amino acids increased with a concomitant reduction in proline and glutamine. Both the barley C-hordein and wheat ω-gliadin genes proved successful for RNAi-gene mediated suppression of barley C-hordein level. All transgenic lines that exhibited a reduction for C-hordein showed off-target effects: the lines exhibited increased level of B/γ-hordein while D-hordein level was reduced. Furthermore, the multicopy insertions correlated negatively with silencing.
OriginalsprogEngelsk
TidsskriftTransgenic Research
Vol/bind25
Nummer1
Sider (fra-til)19-31
Antal sider13
ISSN0962-8819
DOI
StatusUdgivet - feb. 2016

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