In anther cultures ofAnemone canadensisL.,Anemone dichotomaL.,Anemone hupehensisLemoine,Clematis viticellaL. andPapaver setigerumDC. a positive relationship between incubation in 2% CO2and the production of microspore‐derived embryos was observed. In anther cultures ofNicotiana tabacumL.,Anemone hupehensisandClematis viticellaa combination of cold treatment (7°C) and incubation in 2% CO2increased embryo production. InAnemone canadensiscold treatment increased the number of proembryos, whereas incubation in 2% CO2had no effect. InAnemone hupehensis5% CO2increased embryo production by more than 2%. InAnemone dichotomaandPapaver setigerum2% CO2was the more efficient level. CO2had no significant effect on pH in the culture medium in anther cultures ofAnemone canadens