Isolation and characterization of differentially expressed genes in petals of chrysanthemum mutant cultivars developed by irradiation

Abstract

Chrysanthemum is one of the most popular ornamental plants, whose petal colors are primarily determined by pigments including flavonoids/anthocyanins and carotenoids. To develop chrysanthemum cultivars with various petal colors, mutation breeding targeting alteration of pigmentation pattern has been performed. A radiation-induced mutant line, ‘ARTI-purple’, showed a flower color altered from the original bright pink to purple. In this study, we used suppression subtractive hybridization to analyze transcripts and characterize the differential gene expression of chrysanthemum petals between the mutant ‘ARTI-purple’ and its wild-type Chrysanthemum×morifolium cultivar ‘Argus’. One hundred and seventy-six genes were identified (e-value≤1e−5) and classified based on sequence homology to genes with known or putative functions. The genes were categorized functionally by gene ontology analysis and their tentative pathways were confirmed using the TAIR database. The analyses revealed that these genes were related to carbohydrate metabolism, biosynthesis of secondary metabolites, and lipid metabolism. Six genes in a Kyoto encyclopedia of genes and genomes (KEGG) pathway which included the largest number of differentially expressed genes were selected for validation by quantitative PCR, and most of them showed higher expression levels compared with the wild-type. In addition, we isolated two novel clones (PC06E06 and PC08C09) having glutathione S-transferase (GST) family conserved domains and one clone (PC02G08) having a Multidrug and toxic compound extrusion (MATE) family conserved domain based on analysis using conserved domain database (CDD). The expressions of PC08C09 and PC02G08 were upregulated in ‘ARTI-purple’, which implies that anthocyanin accumulation pattern might be altered in mutant. In this study, we identified several differentially expressed genes between ‘Argus’ and ‘ARTI-purple’. The analysis suggested that several metabolic genes as well as glutathione S-transferases and MATEs might be involved in the control of flower pigmentation in chrysanthemum.