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. Author manuscript; available in PMC: 2014 Jul 8.
Published in final edited form as: Curr Biol. 2013 Jun 20;23(13):1209–1214. doi: 10.1016/j.cub.2013.05.021

Figure 2. MYB97, MYB101, and MYB120 are required for pollen tube burst, sperm release and synergid degeneration.

Figure 2

ms1 pistils were pollinated with wild-type LAT52:GFP or myb triple mutant LAT52:GFP pollen 12 hours prior to dissection and imaging by confocal microscopy. (A) Wild-type GFP+ pollen tubes target the micropyle (m) and burst in the female gametophyte yielding a diffuse green signal (arrow). (B) myb triple mutant GFP+ target ovules, overgrow (arrowhead) and fail to burst, with a contained GFP signal inside the pollen tube (inset). Scale bars, 50μm. (C) Quantitative analysis of pollen tube burst by confocal microscopy. The percentage of ovules receiving a pollen tube with a normal burst or a coiling phenotype is shown as a percentage (+/− sd). P-value <0.005, student’s t-test. (D) Wild-type ACT11:MSI1:GFP ovule with position of micropyle (m) and funiculus (fn) indicated. Inset: sn, synergid nucleus; ecn, egg cell nucleus; ccn, central cell nucleus. (E–G) First panel, GFP and DsRed channels merged; middle panel, DsRed channel; last panel, GFP Channel. (E) Wild-type RFP+ sperm nuclei (arrows) undergo double fertilization. (F,G) myb triple mutant RFP+ sperm nuclei remain condensed and associated (arrowheads). (G) Multiple pairs of unfused sperm (arrowheads) were observed myb triple mutant pollinations. Scale bars, 50μm. (H) Quantitative analysis of double fertilization. 1 UFP, ovules with one pair of unfused sperm; 2 UFP, two pairs of unfused sperm; 1 UFP 1 FP, one unfused pair and one fused pair of sperm; 1 FP, one fused pair of sperm. (I–L) Wild-type ACT11:MSI1:GFP pistils were pollinated with wild-type LAT52:DsRed or myb triple mutant, LAT52:DsRed and allowed to grow for 12h. (I) Wild-type, LAT52:DsRed pollen tubes targeting wild-type, ACT11:MSI1:GFP ovules (dsn, degenerated synergid; sn, persisting synergid nucleus), Inset is GFP channel alone. (J) Coiling myb triple pollen tubes with two intact synergid nuclei, (K) one intact synergid or, (L) two degenerated synergids. Scale bars, 50μm. (M) Quantitative analysis of synergid degeneration. Percent of ovules targeted by pollen tubes with; 2 SN, two persisting synergid nuclei; 1 SN, one persisting synergid nucleus; 0 SN, no persisting synergid nuclei; N, total targeted ovules.