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Fig. 3. Partition of gene expression and cell identity within callus.
(A) Mounds of small, dividing cells marked by the
pCUC2::3XVENUS-N7 reporter (green) formed and (B) gave rise to
new shoot meristems (arrowheads), often observed in clusters. Chlorophyll
autofluorescence is in red. (C,D) Scanning electron micrographs
of early regenerating meristems (arrowheads, C), and (D) a late stage
regenerated shoot emerging from callus. (E) The pWUS::mGFP-ER
reporter (green) was expressed in callus cells poorly stained by FM4-64 dye
(red) following 5 days induction on SIM. (F,G) Shoot progenitors
(F, arrowhead, 12 days on SIM) were labeled with FM4-64 dye, and emerged from
regions with peripheral pWUS::mGFP-ER expression, and formed mature
shoot meristems (G), also strongly stained by FM4-64 dye. The
pWUS::mGFP-ER reporter was upregulated in the center of the
developing meristem (arrowhead). (H) pCUC2::3XVENUS-N7 (green)
and pWUS::DsRed-N7 (red) reporters were active in opposing domains of
cells, sometimes in gradients, shown after 10 days on SIM. (I) Higher
magnification after 11 days on SIM, showing clusters of cells expressing the
CUC2 reporter (arrowheads) surrounded by pWUS::DsRed-N7
expressing cells. (J) At later stages, WUS::DsRed-N7
expression was initiated in the center of the mound of shoot progenitors while
pCUC2::3XVENUS-N7 was restricted to the future peripheral zone, shown
here after 12 days on SIM. Scale bars: 50 µm (A,C,I,J); 100 µm (B,E-H);
300 µm (D). Pn, primordia.
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