Fig. 5.

Pollen tube reception is compromised in the mutant of VHA-A. (A and B) Aniline blue staining of wild-type pistil (A) or vha-A/+ pistil (B) emasculated and hand-pollinated with wild-type pollen at 48 HAP. Arrows indicate unfertilized ovules. Two to three overlapping high-magnification images were taken for one pistil. The images were then overlaid with Photoshop (Adobe Systems) to show the whole pistil. (C and D) Representative aniline blue staining of wild-type (C) or vha-A/+ (D) ovules at 48 HAP. Arrowheads point at micropyle in C and a tangled pollen tube in D. Percentage of the displayed category (fertilized in C; defective in pollen tube reception in D) is shown on top right of the image. Results are means ± SEM. Approximately 200 ovules were analyzed in four independent experiments. (E and F) An ovule from WT (E) or vha-A/+ plants (F) pollinated with Pro_LAT52:GFP;Pro_HTR10:HTR10-mRFP pollen at 12 HAP. Images shown on the left are merges of RFP and GFP channel images, whereas on the right are merges of bright-field image and fluorescence image. Dotted line in F indicates the growing pollen tube in the embryo sac. Arrows point at sperm cells. (Scale bars: A and B, 200 µm; C–F, 20 µm.)