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Figure S1. Morphological variation of zygotes within sexual spheroids in a culture of Volvox kirkiorum Nozaki et al. 2006-728-2-B8.

Figure S2. Volvox globator L. SAG 199.80. (A, B) Two views of asexual spheroid. (C) Side view of anterior somatic cells in asexual spheroid. (D, E) Sexual spheroid with eggs (e) and sperm packets (s). (F, G) Two views of zygote.

Figure S3. Volvox barberi W. Shaw UTEX 804. (A, B) Two views of asexual spheroid. (C) Side view of anterior somatic cells in asexual spheroid. (D, E) Sexual spheroid with eggs (e) and sperm packets (s). (F, G) Two views of zygote.

Figure S4. Secondary structures of helices I and IV of ITS-1, and helices I, II, and IV of ITS-2 of Volvox rousseletii G. S. West UTEX 1862, V. ferrisii N. Isaka et al., and V. kirkiorum Nozaki et al. A compensatory base change (CBC) and hemi-CBCs are marked with black and gray backgrounds, respectively. The supposed numbers of CBCs and hemi-CBCs (CBC/hemi-CBC) are shown with the nodes of suggested phylogenetic relationships (see Figs. 3 and 4). Note that the helix III of ITS-1 is missing in the three species.

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