Integrated analysis of miRNA–mRNA regulatory networks in <i>Anoectochilus roxburghii</i> in response to blue laser light

Anoectochilus roxburghii (Wall.) Lind. has significant medicinal and economic value, and the social demand for this species is increasing annually. Laser light sources have different luminescent mechanisms compared with ordinary light sources and are also important factors regulating the synthesis of functional metabolites in A. roxburghii. However, the regulatory mechanism through which A. roxburghii responds to blue laser light has not been investigated. Previous studies have shown that blue-laser treatment results in more red leaves than blue- or white-light treatment. Here, the differences in the effects of laser treatment on A. roxburghii were analyzed by transcriptome sequencing. Gene Ontology analysis revealed that the membranes, calcium ion binding, brassinosteroid-mediated signaling pathway and response to salicylic acid play important roles in the response of A. roxburghii to blue laser light. Kyoto Encyclopedia of Genes and Genomes analysis revealed the involvement of multiple pathways in the response to blue-laser treatment, and among these, beta biosynthesis, flavone and flavonol biosynthesis, thiamine metabolism, limonene and pinene biosynthesis, and peroxisomes play core roles. Cytoscape interaction analysis of the differentially expressed miRNA targets indicated that novel_miR_66, novel_miR_78 and novel_miR_212 were most likely involved in the effect of blue laser light on A. roxburghii. Metabolic content measurements showed that blue laser light increased the beet red pigment, thiamine, total flavonoid and limonene contents, and qPCR analysis confirmed that novel_miR_21, novel_miR_66, novel_miR_188 and novel_miR_194 might participate in the blue-laser signaling network through their target genes and thereby regulate the functional metabolite accumulation in A. roxburghii. This study provides a scientific basis for high-yield A. roxburghii production.