Supplemental Figure S1
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Fig. S1. Genes regulated during callus culture and SAM regeneration. (A) RT-PCR for 18S rRNA or WUS transcript from pCLV3::GFP-ER; pWUS::DsRed-N7 bearing plants. Two-week-old callus samples induced for 9 days on SIM medium were imaged for WUS reporter signal and against CLV3 reporter signal (selecting against mature meristems). WUS transcript was observed in 14 out of 15 samples in which the WUS reporter was observed (boxed lanes). (B) PIN1-GFP signal (green) was upregulated after incubation on CIM (4 days) in the vicinity of callus formation (arrow). Propidium iodide (red). (C) pCUC2::CUC2-VENUS signal (red) was observed in meristem progenitor cells (arrowhead), co-labeled by pPIN1::PIN1-GFP expression (green). (D) After 24 hours of observation, pCUC2::CUC2-VENUS signal (red) was upregulated in the shoot progenitors (arrowhead). pPIN1::PIN1-GFP expression was upregulated in early primordia. (E) After 48 hours, pCUC2::CUC2-VENUS was expressed in the meristem-primordia boundaries. Another cluster of shoot progenitors was initiated nearby (arrowhead). (F) Cells marked by the CUC2 reporter (green) proliferated to form mounds of progenitor cells that were encompassed by cells expressing the WUS reporter (red), which did not divide rapidly. (G) Root meristems (arrowheads), marked by strong pARR5::GFP-ER reporter (green), formed in regions of low ARR5 reporter signal and were stained by propidium iodide (red). (H-S) Same as in Fig. 5J-L but with separate channels for pPIN1::PIN1-GFP (green), pSTM::STM-VENUS (blue) and pWUS::DsRed-N7 (red). Scale bars: 50 μm (B-F,H-S); 100 μm (G). Pn, primordia; In, incipient primordia.
