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. Author manuscript; available in PMC: 2014 Apr 2.
Published in final edited form as: Curr Biol. 2012 May 17;22(12):1090–1094. doi: 10.1016/j.cub.2012.04.041

Figure 1.

Figure 1

Polytubey is rare in wild type, but increases significantly when gamete fusion fails. (A) hap2(gcs1) enhances polytubey. Differentially marked pollen tubes can be tracked as they grow into an ovule at the micropylar end (M). Representative images of ovules targeted by a DsRed+ pollen tube (left), a GFP+ pollen tube (center), or by two different pollen tubes (right). The number of each type of targeting event observed is also shown (bottom). WT, LAT52:GFP pollen mixed with LAT52:DsRed (both types of pollen are wild type) or hap2-1 (hap2-1/HAP2(GCS1), LAT52:GFP mixed with wild-type, LAT52:DsRed pollen). Scale bars, 20μm. (B) duo1 enhances polytubey. duo1-3/+ heterozygous plants were manually self-pollinated and pollen tubes were analyzed 24 hours later using aniline blue staining[14]. Ovules are shown that have attracted a single pollen tube (left panel), two pollen tubes (center panel), or three pollen tubes (right panel). The percentage of ovules with two or more pollen tubes is plotted for No-0 (wild type, 9 pistils, 260 targeted ovules) and duo1-3/+ (8 pistils, 275 ovules targeted). Error bars represent standard deviation (each pistil is one trial). Rates of polytubey are higher in the No-0 accession (4%) than in ms-1 (Landsberg accession, 1%, Figure 1A) and are significantly enhanced by duo1-3 (36%). (C) Multiple hap2-1 sperm are released into a single ovule. The first panel shows an ovule targeted by a wild-type HTR10:HTR10:mRFP, LAT52:GFP pollen tube. Green signal is released from the burst pollen tube; sperm nuclei (SN, red signal) have fused (F) with the egg cell and central cell nuclei. The second panel shows an ovule targeted by a hap2-1, HTR10:HTR10:mRFP, LAT52:GFP pollen tube. The pollen tube has burst (green signal), but sperm cell nuclei remain unfused (UF). The third panel shows an ovule containing four unfused hap2-1, HTR10:HTR10:mRFP sperm. The last panel shows an ovule containing eight unfused hap2-1, HTR10:HTR10:mRFP sperm. Five sperm nuclei are clearly visible, the others are outside the plane of this image. In this ovule, pollen tube cytoplasm (GFP) appears to fill the area occupied by both synergids. Scale bars, 10μm.