Sex-Specific Cell Fusion Pattern of Isogametes in Marine Green Alga, <i>Acetabularia caliculus</i> (Ulvophyceae, Chlorophyta)

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Using field emission scanning electron microscopy (FE-SEM) and fluorescence microscopy, the respective relations between the arrangements of the gamete cell fusion site and the inheritance pattern of chloroplast DNA (cp-DNA) as a sex-specific marker were studied for the isogamous dasycladalean alga Acetabularia caliculus. The gamete had two flagella elongated from the cell anterior. One oval-shaped eyespot situated on the cell posterior was visualized using FE-SEM. When the gametes belonging to the opposite mating types were mixed together, the two gametes aligned side-by-side at their lateral side and fused to form a quadriflagellate planozygote. In the planozygote, the two flagella from the opposite mating type gametes lay side-by-side and two eyespots aligned on the same side of the cell, suggesting opposite positioning of the cell fusion sites between two parental gametes. To confirm whether the gamete fusion pattern observed in the planozygotes was a result of sex-specific fusion or not, the inheritance pattern of cp-DNA was observed using fluorescence microscopy after staining with DAPI concomitantly with the cell fusion pattern. When the eyespots were used as positional marker and were viewed from the surface that included the eyespots, the chloroplast of one parental gamete was found to contain cp-DNAs, although cp-DNAs were not detected in the chloroplast derived from another parental gamete. These results suggest that the gamete fusion pattern observed by FE-SEM was a result of sex-specific fusion.

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