Plant peptide hormone phytosulfokine promotes embryo development of mass in Pinus massoniana

Pinus massoniana is a critical afforestation and ecological tree species in China. However, the continued existence of this pine is severely threatened by pine wilt disease. Somatic embryogenesis serves as a highly efficient clonal propagation approach. Although significant progress has been made in somatic embryogensis research on P. massoniana, resulting in the successful regeneration of plants, the limited embryogenic potential of improved cell lines and loss of embryogenic properties resulting from prolonged proliferation have posed obstacles to the industrialization of SE production. In this study, we investigated the effect of phytosulfokine on embryo development of cell lines from P. massoniana which lead to a cascade of physicochemical changes. Eight embryogenic cell lines of P. massoniana were used to observe phenotype and cytological changes. Physiological factors and the contents of nutrients and endogenous hormones were measured before and after phytosulfokine addition. We found that PSK promoted a change in the embryogenic mass of P. massoniana, leading to their development from pro-embryogenic mass (PEM)I to PEMII or PEMIII stages of pro-embryos. In addition, PSK accumulated soluble sugar, protein, and starch, and maintained redox homeostasis during cell line proliferation by reducing H₂O₂ levels. Our findings increase our understanding of how PSK affects somatic embryogensis in P. massoniana, thereby providing a valuable tool for establishing efficient somatic embryogensis systems in conifer species.