Common buckwheat (Fagopyrum esculentum) is a short-season grain crop that is a source of rutin and other phenolic compounds. In this study, we isolated the cDNAs of 11 F. esculentum enzymes in the flavonoid biosynthesis pathway, namely, phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate:CoA ligase (4CL) 1 and 2, chalcone synthase (CHS), chalcone isomerase (CHI), flavone 3-hydroxylase (F3H), flavonoid 3'-hydroxylase (F3'H), flavonol synthase (FLS) 1 and 2, and anthocyanidin synthase (ANS). Quantitative real-time polymerase chain reaction analysis showed that these genes were most highly expressed in the stems and roots. However, high performance liquid chromatography analysis indicated that their flavonoid products, such as rutin and catechin, accumulated in the flowers and leaves. These results suggested that flavonoids may be transported within F. esculentum. In addition, light and dark growth conditions affected the expression levels of the biosynthesis genes and accumulation of phenolic compounds in F. esculentum sprouts.

Radish [Raphanus sativus (Rs)] is an important dietary vegetable in Asian countries, especially China, Japan, and Korea. To elucidate the molecular mechanisms of anthocyanin accumulation in radish, the gene expression of enzymes directly involved in anthocyanin biosynthesis was analyzed. These genes include phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol reductase (DFR), and anthocyanidin synthase (ANS). RsDFR and RsANS were found to accumulate in the flesh or skin of two radish cultivars (Man Tang Hong and Hong Feng No.1). Radish skin contained higher CHS, CHI, and F3H transcript levels than radish flesh in all three cultivars. In the red radish, 16 anthocyanins were separated and identified by high-performance liquid chromatography (HPLC) and elctrospray ionization-tandem mass spectrometry (ESI-MS/MS). Some of them were acylated with coumaroyl, malonoyl, feruoyl, and caffeoyl moieties. Furthermore (-)-epicatechin and ferulic acid were also identified in the three cultivars.

Buckwheat sprouts are considered an excellent dietary source of phenolic compounds. The time duration and amount of light for sprouting strongly affect the nutritional quality of sprouts. In this study, these two factors were investigated in two cultivars of tartary buckwheat sprouts: Hokkai T8 and T10. The transcriptional levels of flavonoid biosynthetic genes were investigated in light/dark- and dark-treated sprouts. Among the main flavonoid biosynthesis structural genes, FtPAL, Ft4CL, FtF3H, FtDFR, and FtANS exhibited higher transcriptional levels than others as compared to that of a housekeeping gene (histone H3) during sprouting; FtF3'H1, FtF3'H2, FtFLS2, and FtANS were substantially upregulated at 2, 4, and 6 days in light/dark-treated T10 sprouts than in dark-treated ones. However, FtDFR was downregulated in 8 and 10 day old light/dark-treated sprouts of both cultivars. High-performance liquid chromatography (HPLC) analysis revealed that increasing the culture time did not affect the accumulation of flavonoids or anthocyanins. However, light contributed the production of anthocyanins in Hokkai T10 sprouts. The anthocyanins included cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, and delphinidin-3-O-coumarylglucoside, which were identified by HPLC and electrospray ionization-tandem mass spectrometry. Instead of anthocyanins, Hokkai T8 sprouts produced large amounts of 4 flavonoid C-glycosylflavone compounds in both light/dark and dark conditions: orientin, isoorientin, vitexin, and isovitexin. These results indicate that these two types of tartary buckwheat sprouts have different mechanisms for flavonoid and anthocyanin biosynthesis that also vary in light/dark and dark conditions.

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Korean mint (Agastache rugosa), a perennial, medicinal plant of the Labiatae family, has many useful constituents, including monoterpenes and phenylpropanoids. Among these, tilianin and rosmarinic acid, 2 well-known natural products, have many pharmacologically useful properties. Chalcone synthase (CHS) and chalcone isomerase (CHI) catalyze the first and second committed steps in the phenylpropanoid pathway of plants, leading to the production of tilianin. In this study, cDNAs encoding CHS (ArCHS) and CHI (ArCHI) were isolated from A. rugosa using rapid amplification of cDNA ends (RACE)-PCR. Amino acid sequence alignments showed that ArCHS and ArCHI shared high sequence identity and active sites with their respective orthologous genes. Quantitative real-time PCR analysis was used to determine the expression levels of genes involved in tilianin and rosmarinic acid biosyntheses in the flowers, leaves, stems, and roots of A. rugosa. High-performance liquid chromatography (HPLC) revealed that the accumulation pattern of tilianin matched the expression patterns of ArCHS and ArCHI in different organs of A. rugosa. Moreover, acacetin, the precursor of tilianin, also demonstrated an accumulation pattern congruent with the expression of these 2 genes. The transcription levels of ArPAL, ArC4H, and Ar4CL were the highest in the leaves or flowers of the plant, which also contained a relatively high amount of rosmarinic acid. However, the roots showed a significant content of rosmarinic acid, although the transcription of ArPAL, ArC4H, and Ar4CL were low. The findings of our study support the medicinal usefulness of A. rugosa and indicate targets for increasing tilianin and rosmarinic acid production in this plant.