Summary Different colors, such as purple, brown, red and white, occur in the pericarp of rice. Here, two genes affecting proanthocyanidin synthesis in red‐ and brown‐colored rice were elucidated. Genetic segregation analysis suggested that the Rd and A loci are identical, and both encode dihydroflavonol‐4‐reductase (DFR). The introduction of the DFR gene into an Rcrd mutant resulted in red‐colored rice, which was brown in the original mutant, demonstrating that the Rd locus encodes the DFR protein. Accumulation of proanthocyanidins was observed in the transformants by the introduction of the Rd gene into the rice Rcrd line. Protein blot analysis showed that the DFR gene was translated in seeds with alternative translation initiation. A search for the Rc gene, which encodes a transacting regulatory factor, was conducted using available DNA markers and the Rice Genome Automated Annotation System program. Three candidate genes were identified and cloned from a rice RcRd line and subsequently introduced into a rice rcrd line. Brown‐colored seeds were obtained from transgenic plants by the introduction of a gene containing the basic helix–loop–helix (bHLH) motif, demonstrating that the Rc gene encodes a bHLH protein. Comparison of the Rc locus among rice accessions showed that a 14‐bp deletion occurred only in the rc locus.

The 80% ethanol extracts of 5 purple-fleshed sweet potato cultivars were separated into 2 fractions, anthocyanins-and phenolic compounds-rich fractions, to clarify the contribution of these constituents to the radical-scavenging activity. The separation was accomplished with an ethyl acetate liquid/liquid extraction. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity in each fraction and the contributors varied according to the cultivars. The dominant DPPH radical-scavengers in "Ayamurasaki" and "Kyushu-132" were anthocyanins rather than phenolic compounds, while those in "Miyanou-36" and "Bise" were phenolic compounds, such as chlorogenic acid. Furthermore, the high-performance liquid chromatography analysis of anthocyanins showed that "Ayamurasaki" and "Kyushu-132" were rich in anthocyanins with peonidin aglycon, whereas "Miyanou-36," "Bise," and "Tanegashimamurasaki" contained cyanidin aglycon.

The extracts from white-, black-, and red-hulled rice were prepared by sequential extraction with six different polar solvents, and their radical-scavenging activities were measured by methods using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH*) and tert-butyl hydroperoxyl radical (t-BuOO*). The extracts prepared with highly polar solvents, methanol and deionized water, exhibited higher DPPH* and t-BuOO* scavenging activities in all three cultivars. In addition, the acetone extract from red-hulled rice exhibited a high DPPH* and t-BuOO* scavenging activity, while no such activity was detected for the acetone extracts from white- and black-hulled rice. The major components responsible for the radical scavenging in the acetone extract from red-hulled rice were identified as procyanidins by acidic hydrolysis, vanillin assay, and Sephadex LH-20 chromatography. GPC analysis of the acetylated procyanidins revealed that the average molecular weight is about 5000, in a range of about 500-18,000.

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1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity of the 70% aqueous acetone extract from the seed coat of the brown soybean variety, Akita-Zairai, was investigated. The activity of the seed coat of Akita-Zairai was much higher than that of three other reddish-brown varieties, but lower than that of two black varieties, and was closely dependent on the content of phenolic compounds. In the LH20 column chromatography of Akita-Zairai, high DPPH radical-scavenging activities were detected in the fractions eluted with MeOH and 70% aqueous acetone. Proanthocyanidins were also detected in fractions showing high radical-scavenging activities. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis showed that the degree of polymerization (DP) of the procyanidins contained in the brown or black soybean seed coat was as high as DP30.

Absorption of acylated anthocyanins in purple-fleshed sweet potato (Ipomoea batatas cv. Ayamurasaki) in rats was studied to obtain evidence that the acylated anthocyanins themselves could exert a physiological function in vivo. Peonidin 3-caffeoylsophoroside-5-glucoside (Pn 3-Caf‚sop-5-glc) in purple-fleshed sweet potato was directly absorbed into rat and present as an intact acylated form in plasma. After oral administration of the purple-fleshed sweet potato anthocyanin (PSA) concentrate containing 38.9 µmol of Pn 3-Caf‚sop-5-glc/kg of body weight, Pn 3-Caf‚sop-5-glc was detected in the plasma, and the C max value and t max were estimated as 50.0 ( 6.8 nmol/Lof plasma and 30 min, respectively. Furthermore, the plasma antioxidant capacity was significantly elevated from 58.0 ( 12.0 to 89.2 ( 6.8 µmol of Trolox equivalent/L of plasma 30 min after the administration of the PSA concentrate.

Stabilization of the levels of active oxygen species (AOS) is important to the survival of organisms. To clarify the system controlling levels of AOS in plants, this study used an electron spin resonance (ESR) method to directly measure superoxide radical (O(2)(.-)) scavenging activities in the wild-type Arabidopsis thaliana (Col and Ler ecotypes), two anthocyanin mutants (tt3 and ttg1), and an ascorbic acid mutant (vtc1). Under ordinary growth conditions, Arabidopsis contained superoxide-scavenging activity (SOSA) of approximately 300-500 SOD units/g of fresh weight. The ESR pattern indicated that most (40-50%) of this activity was due to ascorbic acid. For the analysis of SOSA under conditions of oxidative stress, synthesis of AOS was induced by gamma-irradiation. The radical scavenging activity in irradiated plants increased approximately 10-fold following an associated increase in the accumulation of ascorbic acid and anthocyanin. The accumulation of ascorbic acid and anthocyanin was suppressed by treatment with an antioxidant before irradiation and was induced by treatment with a radical-generating reagent. The contributions of ascorbic acid and anthocyanin to the total superoxide radical scavenging activity differed among ecotypes. In the Ler ecotype, ascorbic acid accumulated at twice the level of that in the Col ecotype, and induction of anthocyanin was half that in Col. To confirm the activity of ascorbic acid and anthocyanin against AOS stress, the viability of the wild type and mutants (tt2, tt3,tt5, ttg1, and vtc1) was examined after gamma-irradiation. Only the plants in which ascorbic acid and anthocyanin were induced had the ability to grow and flower.