Karyogamy in rice zygotes: Actin filament-dependent migration of sperm nucleus, chromatin dynamics, and de novo gene expression

Abstract

In angiosperms, the fusion of a sperm cell with an egg cell, termed plasmogamy, triggers egg activation. Then, karyogamy, migration of the sperm nucleus toward the egg nucleus and their subsequent nuclear fusion, progresses, and de novo gene expression from the zygotic genome is initiated for early embryogenesis. Therefore, karyogamy is an important post-fusion event that bridges egg activation and de novo gene expression in fused gametes/zygotes. In this study, we monitored the progression of karyogamy in rice zygotes produced by in vitro fusion. The results indicated that the sperm nucleus migrated adjacent to the egg nucleus via an actin cytoskeleton, and the egg chromatin then appeared to move unidirectionally into the sperm nucleus through a possible nuclear connection. An enlargement of the sperm nucleus accompanied this possible chromatin remodeling. Then, 30–70 min after fusion, the sperm chromatin began to decondense, and karyogamy was completed. The development of early rice zygotes from plasmogamy to karyogamy could be divided into eight stages, and paternal and de novo synthesized transcripts were separately detectable in zygotes at early and late karyogamy stages, respectively, by RT-PCR using zygotes at each karyogamy stage.