Extended Data Fig. 6: Over-expression of DAZ1 and DAZ2 in sperm cells does not affect gamete fusion.
From: EGG CELL 1 contributes to egg-cell-dependent preferential fertilization in Arabidopsis
a-b, qPCR analyses of DAZ1 (a) and DAZ2 (b) in opening flowers of plants expressing pDMP9::DAZ1 or pDAF1::DAZ2. The expression level of DAZ1 and DAZ2 in the wild type (Col-0) was set to 1. Data are the means ± SD from three independent experiments. c-d, Quantitative analyses of pollen containing two sperm cells in different pDMP9::DAZ1 and pDAF1::DAZ2 lines. The number of sperm cells marked by HTR10-RFP was counted in mature pollen grains (from left to right, n = 502, 509, 445, 456, 493, 535 and 439, respectively). e, Representative images of gamete fusion events in WT ovules 8-10 hours after pollination with WT, pDMP9::DAZ1 or pDAF1::DAZ2 pollen. The decondensed sperm chromatin (arrowheads) was taken as a sign of gamete fusion. Arrows indicate pollen tube contents (labeled by pLAT52::GFP). f, Quantitative analyses of double or single fertilization 8-10 HAP in WT ovules, when pollinated with WT, pDMP9::DAZ1 or pDAF1::DAZ2 pollen. From left to right, n = 166, 143, 146, 167, 190, 173 and 156 ovules. g, Seed morphology in self-crossed siliques in wild type, pDMP9::DAZ1 and pDAF1::DAZ2 lines. h, Quantification of developed seeds shown in g (n = 10 siliques for each). From left to right, the total number of seeds is 584, 568, 548, 590, 559, 677 and 640, respectively. In a, b, d, f and h, data are the means ± SD and significant differences by two-sided Student’s t-test are indicated. Scale bars, 10 µm in c, e and 0.5 mm in g. Abbreviations: CC, central cell; EC, egg cell.
