Ovary development and polydnavirus morphogenesis in the parasitic wasp Chelonus inanitus. II. Ultrastructural analysis of calyx cell development, virion formation and release.

Polydnaviruses are unique symbiotic viruses that are formed only in calyx cells in the ovary of parasitic wasps in the families Braconidae and Ichneumonidae; accordingly, two genera, Bracovirus and Ichnovirus are recognized. We have presented a detailed ultrastructural analysis of ovary and calyx cell differentiation and virion morphogenesis, together with the first data on virion release in a bracovirus. Differentiation of the ovary into germarium/vitellarium and the calyx region begins immediately after pupation. In the periphery and central part of the calyx, some cells and their nuclei begin to enlarge and the DNA content increases. The calyx cell nuclei then further increase and become highly lobulated, nuclear pores become very abundant and the cytoplasm is rich in ribosomes. This suggests synthesis and import of viral envelope proteins as viral envelopes appear in the nuclei shortly later. The appearance of viral envelopes is accompanied by a swelling of the nucleus and a change in electron density. Thereafter, the calyx cells reach the final stage with a highly swollen nucleus containing virogenic stroma and mature virions with nucleocapsids. Up to this stage, the DNA content of nuclei increases 120-fold and the volume 45-fold. The mature calyx cells are positioned in the vicinity of the oviduct lumen; for release of virions first the nuclear and then the plasma membrane disintegrate. On the border of the oviduct lumen, cells of an epithelial layer become phagocytic and remove debris, leading to a calyx fluid that contains only densely packed virions.