Abstract: In our study, a complete open reading frame of PyMFG was obtained by RT-PCR based on the sequence of differentially expressed genes (Contig-21827) screened by transcriptome sequencing of Pyropia yezoensis. Sequence analysis showed that the ORF of the PyMFG was 1 224 bp in length and encoded a polypeptide fragment of 407 amino acids with a molecular weight of 46.24 ku, and theoretical pI of 9.08. Domain analysis revealed that the protein contains a conserved TEA domain and a YAP domain, which belongs to the TEA-ATTS superfamily domain. We found that the protein and fungal conidia forming proteins clustered into one large branch with close genetic relationship through multiple sequence alignment and phylogenetic analysis. In addition, the comparison of the ability of releasing monospores and qRT-PCR analysis between different strains indicated that the expression trend of PyMFG in PY26W and PY26W' was highly consistent with macroscopic statistical results, while the trend in PY26R and PY26R' were exactly existed difference. By inference, the gene type and molecular mechanism regulating the formation and release of monospores in the parental (PY26W) and partial parental strain (PY26W') and the female parental (PY26R) as well as partial female strain (PY26R') existed certain difference, leading to the divergence between the macroscopic statistical results of monospores and the qRT-PCR results.