3 research outputs found
Characterization of MT1 (128/Xyl, cf. Fig. S1e) transformants.
<p>(A) PCR analysis of 13 randomly chosen doubled-haploid T<sub>1</sub> seedlings of primary transformant MT1 B4. Lanes: 1β=β1 kb ladder; 2β=β plasmid DNA; 3β=β wild type wheat DNA; 4β16β=βT<sub>1</sub> seeds. (B) Southern blot analysis of homozygous MT1 transformants from T<sub>1</sub> seedlings with the xylanase gene. Lanes: C1, C2, C4 respectively showing 2, 4 and 8 copies of 837 bp probe; Mβ=β DNA ladder; lanes 1β10β=βT<sub>1</sub> doubled-haploid seedlings of 6 different T<sub>0</sub> transformants (1 and 2β=βMT1-1, 3 and 4β=βMT1-2, 5 and 6β=βMT1-3, 7β=βMT1B5, 8 and 9β=βMT1-6, and 10β=βMT1-7); lanes 11β13β=βT<sub>1</sub> doubled-haploid seedlings of MT1-B4; lanes 14β15β=β wild type DNA. (C) Zymogram assay for identification of transgenic wheat grains synthesizing recombinant 1,4-Ξ²-xylanase. Transgenic wheat grains (T<sub>2</sub> of MT1-B4) secrete the enzyme into the medium containing oat-spelt xylan that is stainable with Congo Red. De-polymerization of the xylan by the enzyme results in an unstained yellow ring around the seed. Wild type wheat grains lack the yellow ring (arrows).</p
Electron micrographs of wheat microspores after pretreatment using transmission electron microscopy.
<p>Figures a-c show differences in thickness of intine, number of cytoplasmic organelles and amount of starch accumulated in amyloplasts. (A) represents type I developmental pathway, (B) represents type II developmental pathway, and (C) represents type III developmental pathway. amβ=β amyloplast, exβ=β exine wall, inβ=β intine layer, mtβ=β mitochondria, glβ=β Golgi apparatus, pβ=β proplastid, rerβ=β rough endoplasmic reticulum, stβ=β starch.</p
Developmental pathways of pre-treated wheat microspores in culture, as determined by time-lapse tracking.
<p>According to the type of development, microspores are grouped into three classes: type I (AβH), type II (IβO) and type III (PβR). Microspores were stained with FM 4β64 (Molecular Probes Cat. # T-3166) to confirm their viability in culture. All pictures were taken at a 25Γ magnification and 6Γ optical zoom except for G, O and H, where the former two pictures were taken at the same magnification but at 3Γ optical zoom and the latter picture was taken at 10Γ magnification and 1.7Γ optical zoom.</p