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In a preceding paper (Briza, P., Winkler, G., Kalchhauser, H., and Breitenbach, M. (1986) J. Biol. Chem. 261, 4288-4294), we reported the presence of dityrosine in the outer layers of yeast ascospore walls. Both outer layers seen in electron micrographs of yeast ascospore walls are sporulation-specific. Here we show that the second of these two outer layers consists of chitosan. In intact spores, it is shielded from staining with primulin by the outermost layer. However, in purified sporepmid:3042773 fatcat:ad77z7ucx5gjfl66ky62xmwhhm