Metabolome and transcriptome analyses of the flavonoid biosynthetic pathway for the efficient accumulation of anthocyanins and other flavonoids in a new duckweed variety (68-red)

Duckweed is a kind of aquatic plant with the characteristics of high nutritional value and medicinal benefits. However, most researches focused on the natural germplasms. The underlying metabolic pathway remains to be systematically elaborated in duckweed. In our laboratory, one reddish-purple mutant with high-flavonoids was screened from a mutant library of Spirodela polyrhiza 6068, named 68-red. The content of anthocyanins and proanthocyanidins in 68-red mutant increased by 563.47% and 231.19%, respectively, compared to wild type. It is interesting that cynaroside and orientin content were significantly increased, in contrast, apigetrin and vitexin were decreased in 68-red mutant. Considering this, metabolome and transcriptome were employed to explore the flavonoids biosynthetic pathway. Here, a total of 734 metabolites were identified in the wild type and 68-red mutant. Among which, cyanidin-3-O-glucoside, cyanidin-3-O-galactoside, pelargonidin-3-O-glucoside and pelargonidin-3-O-(6″-O-malonyl)glucoside were significantly accumulated, which were positively correlated with deep reddish-purple of 68-red mutant. In addition, proanthocyanidins (B1, B2, B3, B4, C1, C2), flavonoid and its glycosides (11 luteolin and its glycosides, 14 quercetin and its glycosides, 14 kaempferol and its glycosides, 2 apigenin glycosides) were significantly accumulated, 2 apigenin glycosides were down-regulated in 68-red mutant. The transcriptome data and qRT-PCR indicated that 16 enzyme genes in flavonoids biosynthetic pathway (PAL, C4H, CHSs, F3H, ANS, ANR, F3'Hs, DFRs, LAR, GT1, BZ1) were significantly up-regulated in 68-red mutant. Correlation analysis found that three copies of F3'H gene play important roles in the synthesis of anthocyanins, luteolin and apigenin glycosides. In conclusion, the 68-red mutant is a high quality germplasm resources for food and medical industry. Metabolome and transcriptome provide new insight for exploring the enzyme genes and functional metabolites in duckweed.