Physiological and Transcriptomic Analysis of a Sepal Mutant in Phalaenopsis

MADS-box transcription factors have undergone in-depth investigations regarding their function in regulating the development of plant floral organs. Flower type mutants serve as critical biological models for investigating the regulatory mechanisms of MADS-box genes in floral organ development, while simultaneously constituting essential genetic resources for molecular breeding programs. In this work, we examined a lip-like sepal of the peloric mutant in Phalaenopsis ‘Huayang’, which exhibited changes in both the morphology and color of the sepals. Our cryo-SEM investigations revealed that the mutation type belonged to a sepal labellum-like variation in Phalaenopsis ‘Huayang’. Nine glycosylated anthocyanins were identified and their contents were significantly upregulated in the Se-red of mutant flowers. Transcriptomic analysis identified 9408 differentially expressed genes, including 4934 upregulated and 4474 downregulated genes. In addition, 57 MADS-box genes were identified and classed into five groups (Mα, Mβ, Mγ, MIKC*, and MIKCC) according to a phylogenetic comparison with Arabidopsis homologs. Furthermore, 29 MADS genes were screened from the MIKCC group, and these genes may play a crucial role in the regulation of floral organ development. Through real-time PCR analysis and protein interaction analysis, we identified three genes that were upregulated in the mutant, which may be involved in sepal development. The subcellular localization results demonstrated that three genes were found within the nucleus. Taken together, our results elucidated the molecular mechanism of sepal variation in Phalaenopsis ‘Huayang’. Our results could enhance our comprehension of the regulatory mechanisms underlying floral patterning and promote the molecular breeding process of Phalaenopsis.