The shedding of gametes of Botryllus primigenus Oka occurs in the natural environment early in the morning, about 1 h after dawn. The shedding of the sperm from a testis is completed within a few seconds. All the spermatozoa in a testis are released at once. Ovulation begins about 10 min after shedding of sperm and is completed in about 1 min. In general, combinative testes, i.e., testes accompanied by egg follicles, macure first, followed by egg follicles, solitary testes located on the left side of the zooid, and finally those on the right side, in that order. As a result, a population of zooids of the same generation in a colony sheds sperm on 2 or 3 successive days. Illumination preceded by darkness effectively induces shedding of gametes experimentally. The required minimum dark-adaptation period is 40 min; the required minimum illumination period is dependent on light-intensity, 8, 4 and 1 min being required at 70, 200 and 3,000 lux, respectively; the latent period is temperature-dependent, 100 to 130, 70 to 90 and 50 to 60 min being required at 21o, 26o and 27.5°C, respectively. An experiment in which only part of the colony was illuminated clearly indicated that each zooid responds to light-stimulus independently.