Isolation and characterization of a spore development mutant in <i>Pyropia yezoensis</i> (Rhodophyta) induced via insertional mutagenesis

Mechanistic elucidation of spore development is essential for understanding reproductive strategies and improving seaweed cultivation. In this study, we isolated a mutant with spore development defects in Pyropia yezoensis (Rhodophyta) from among genetic transformants generated via insertional mutagenesis. We characterized the mutant phenotype and identified the genomic region containing the mutation. The mutant produced spores (archeospores, zygotospores, and conchospores) at every relevant stage of the life history. No evident differences were observed in the appearance of the released spores at any stage under light microscopy between the wild-type and mutant strains; however, the mutant exhibited a trend of reduced spore release compared with the wild-type. Additionally, the archeospores released by the mutant remained round after being released from the sporangium, failed to adhere to the substratum, did not germinate, and showed no cell wall development after 4 days of standard culture, in contrast to the wild-type. Similar results were observed for the zygotospores and conchospores. These findings indicate that the main characteristic of this mutant was poor spore release, and the released spores failed to germinate. Whole-genome sequencing revealed the introduction of exogenous DNA into the nuclear genome of the mutant and suggested the presence of duplications, deletion, and inversion in the mutant genome. Notably, a deletion of approximately 13 kb was detected in the mutant genome flanking the insertion, and this region contained a plant homeodomain, suggesting that homeodomain-containing proteins may regulate spore development in P. yezoensis.