Transcriptomic and protein-protein interactions network analyses of the molecular mechanisms underlying the mycorrhizal interaction in Cypripedium macranthos var. rebunense

Chunyi Jin¹Kota Kambara²Kaien Fujino¹Hanako Shimura^1*

• ¹Graduate School of Agriculture, Hokkaido University, Sapporo, Japan
• ²Asian Research Center for Bioresource and Environmental Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, Japan

To comprehensively analyze the gene expression associated with early fungal colonization, transcriptome analysis of Cypripedium macranthos var. rebunense was performed using mycorrhizal tissues prepared by inoculating seedling plants with a fungus that exhibited different mycorrhizal interaction properties among subcultures.Colonization with the mycorrhizal fungus induced an increased expression of orchid genes encoding enzymes involved in cell wall synthesis, degradation, and modification, as well as those encoding transporters of sugars, amino acids, nucleic acids, and other nitrogen-containing compounds. Enrichment analysis focusing on genes associated with protein-protein interactions (PPI) suggested a potential role of lectin domain-containing receptorlike kinases (LecRLK) in the recognition of fungal colonization and induction of cell wall-modifying enzymes and nutrient transporters required for mycorrhizal formation. Kinase genes such as MAPKKK and serine/threonine protein kinase were upregulated in tissues exhibiting continued peloton formation, whereas these genes exhibited no changes in tissues showing no peloton formation 4 weeks after inoculation. These results suggest that the continuous phosphorylation signaling cascade plays a crucial role in the regulatory pathway for maintaining mycorrhizal interactions between Cypripedium and its mycorrhizal fungus.