Genetic polymorphisms in Japanese fragrant landraces and novel fragrant allele domesticated in northern Japan.

Rice fragrance is an important characteristic for Southeast Asian consumers, and fragrant landraces from Japan were first recorded in the 17th century. Principal component analysis clearly showed that Japanese fragrant landraces were genetically different from non-Japanese fragrant landraces. Japanese fragrant landraces were composed of six clades, none of which carried the most common fragrance mutation, an 8-bp deletion in exon 7 of Badh2. Fragrant landraces comprised two major groups carrying different Badh2 mutations. One group carried a known SNP at exon13 and the other a SNP at the exon1-intron1 junction as splicing donor site. The latter was considered to be a potential splicing mutant group as a novel allele at Badh2. Heterozygosity (He) scores in the two fragrant groups were not significantly different from non-fragrant landraces and modern cultivars. However, lower He scores were found around the Badh2 locus in the two groups. The potential splicing mutant group showed a more extended haplotype than the E13 SNP group. A likely causal factor responsible for loss of function is a novel splicing mutation allele that may have been generated quite recently. The fragrance allele has dispersed as a result of out-crossing under local environmental conditions.

Physiological factors affecting transcription of genes involved in the flavonoid biosynthetic pathway in different rice varieties.

Flavonoids play an important role in the grain color and flavor of rice. Since their characterization in maize, the flavonoid biosynthetic genes have been extensively studied in grape, Arabidopsis, and Petunia. However, we are still a long way from understanding the molecular features and mechanisms underlying the flavonoid biosynthetic pathway. The present study was undertaken to understand the physiological factors affecting the transcription and regulation of these genes. We report that the expression of CHI, CHS, DFR, LAR, and ANS, the 5 flavonoid biosynthetic genes in different rice varieties, differ dramatically with respect to the stage of development, white light, and sugar concentrations. We further demonstrate that white light could induce the transcription of the entire flavonoid biosynthetic gene pathway; however, differences were observed in the degrees of sensitivity and the required illumination time. Our study provides valuable insights into understanding the regulation of the flavonoid biosynthetic pathway.

Anti-oxidative analysis, and identification and quantification of anthocyanin pigments in different coloured rice.

Anthocyanin pigments in coloured rice cultivars were isolated and identified using high-performance liquid chromatography techniques. Two black rice cultivars (Asamurasaki, Okunomurasaki) contained three major anthocyanins: cyanidin-3-glucoside, peonidin-3-glucoside and malvidin. Chinakuromai (black) rice additionally contained a fourth anthocyanin, petunidin-3-glucoside. Four red rice cultivars contained only malvidin. The total anthocyanin content varied greatly among black rice cultivars (79.5-473.7 mg/100g), but was lower in red rice (7.9-34.4 mg/100g). Total phenolic content was similar between red (460.32-725.69 mg/100g) and black (417.11-687.24 mg/100g) rice. The oxygen radical absorbing capacity was ranked as follows: red (69.91-130.32 µmol Trolox/g)>black (55.49-64.85 µmol Trolox/g)>green (35.32 µmol Trolox/g)>white (21.81 µmol Trolox/g) rice. The antioxidant capacity resulted mainly from the seed capsule, not the endosperm. The anthocyanin pigments contributed little to the total antioxidant capacity of red (0.03-0.1%) and black (0.5-2.5%) rice cultivars. Hence, the antioxidant capacity is derived mainly from other phenolic compounds.

A flow-through electrochemiluminescence sensor for salt stress on rice plants.

Cultivation of rice plants (Koshihikari, IR4595 and IR28) in 250 mmol/L NaCl over 16 days provided interesting profiles of responses on a flow-through electrochemiluminescence (ECL) system with tris(2,2'-bipyridine)ruthenium complex. Although some time lags were noted among these rice plants, the increment in the ECL responses was clearly observed on the 11th day. The obtained ECL response profiles were consistent with the proline profiles obtained by HPLC. Such consistency demonstrated that the present ECL system would find a utility as a useful sensor for salt stress on rice plants.

Glucosylation of phenolic compounds by Pharbitis nil hairy roots: I. Glucosylation of coumarin and flavone derivatives.

Hairy roots of medicinal morning glory (Pharbitis nil) showed potent glucosylation activity against umbelliferone and aesculetin, so the glucosylation activity against several phenolic compounds was tested. Some coumarin derivatives and flavone derivatives having phenolic hydroxyl groups were incubated with the hairy roots. The coumarin derivatives and flavone derivatives almost disappeared from the culture medium in half a day. In the case of the coumarin derivatives, a 7-hydroxyl group was easily glucosylated. A methyl group at C-8 somewhat decreased the glucosylation to a hydroxyl group at C-7 of the coumarin skeleton. The 4-hydroxy coumarin derivatives were changed to acetophenone-type glucosides by incubation with the hairy roots through decarboxylation. Several flavonol derivatives were tested for glucosylation by the hairy roots. 3-Hydroxy flavone, 3.6-dihydroxyflavone and 3,7-dihydroxyflavone were glucosylated to give 3-glucosylated derivatives. Of these, 3,6-dihydroxyflavone was highly glucosylated, but not 3-hydroxyflavone or 3,7-dihydroxyflavone to the same degree. In the case of the flavones, a 3-hydroxy group could be predominantly glucosylated, and hydroxyl groups on the A and B ring of the flavones affected glucosylation by the hairy roots.

Umbelliferone released from hairy root cultures of Pharbitis nil treated with copper sulfate and its subsequent glucosylation.

Hairy root cultures of Pharbitis nil treated with CuSO4 and methyl jasmonate (MeJA) produced umbelliferone (1) and scopoletin (2) in the culture medium, and skimmin (3), a beta-D-glucopyranoside of 1, was isolated from the hairy roots. While 1 in the medium increased and reached a maximal level 16 h after the treatment with CuSO4, the amount of 3 in the hairy roots decreased, reaching a minimal level after 8 h, before recovering to a level higher than the basal level after 24 h and then continuously increasing. These observations suggest that 1 was released by the hydrolysis of 3. Umbelliferone (1) inhibited hairy root growth, while skimmin (3) did not. This result suggests that, after the release of 1 as a phytoalexin, the hairy roots glycosylated 1 for the detoxification and re-use of 3 as a source of phytoalexin.

Partitioning of 13C-labelled photosynthate varies with growth stage and panicle size in high-yielding rice.

A super-high-yielding rice (Oryza sativa L.) cultivar, Takanari, and a traditional japonica rice cultivar, Nakateshinsenbon, were grown under field conditions to compare partitioning of 13C-labelled photosynthate to different plant organs during the period of reproductive development. The flag leaf and the two leaves immediately below it on the main culm were exposed individually to 13CO2 and the movement of the heavy carbon isotope to grains, hull, panicle branches and vegetative parts of plant was assessed. Also, the effect of a reduction of sink size on the partitioning of 13C to different organs was studied by removing some of the primary branches of the panicle. 13C taken up by the three leaves in the post-heading period, moved mostly to the grains and hull of the panicle. At this stage, the uppermost three leaves and the panicle consisted of a single source-sink unit. Partitioning of 13C to the rest of the vegetative structures of the plant was minimal. In the case of Nakateshinsenbon, the flag leaf supplied most of the carbon assimilates for the grains and contributions from the other two leaves were much smaller. However, in Takanari, the contribution of 13C to grains from the second leaf was equivalent to that of the flag leaf. In Takanari, removal of more than one third of the primary branches of the panicle significantly reduced partitioning from the third leaf of the culm, but partitioning from the flag leaf was not significantly changed. In contrast, branch removal treatment significantly depressed transport of carbon assimilates from the flag leaf in Nakateshinsenbon. The obligatory nature of the source-sink relationship in rice is discussed. It is concluded that in lower-yielding traditional rice, photosynthesis in the flag leaf supplies carbon assimilates to the developing grains. But in the super-yielding rice Takanari, the main source area is extended to include the two leaves below the flag leaf so as to sustain an extra large panicle. Even greater grain-filling is possible in super-yielding rice, if the source area is increased further.

Distribution of amylose, nitrogen, and minerals in rice kernels with various characters.

The distribution of chemical constituents is known not to be even within a rice kernel. To improve the eating quality of rice or to give it some special traits by adjusting the milling intensity, we investigated the distribution of amylose, nitrogen (N), and specific minerals (P, K, Mg, Ca, and Mn) in rice kernels of 11 cultivars with various characteristics cultivated under similar conditions. The distributions of these constituents were determined using flour samples prepared consecutively by abrasive milling from the outer to the inner portions of hulled rice. In all the cultivars tested, N and the minerals were found to be more abundant in the outer than in the inner portion, but amylose was rich in the inner portion. P, Mg, K, and Mn were extremely rich in the outer portion, while N and Ca were only relatively rich there. Koshihikari, which is the most popular cultivar in Japan because of its excellent eating quality, showed the highest Mg/K ratio in the outermost portion of polished rice. The color of flour samples became pure white going from outside portions toward the center of the kernel, even if the sample was from red rice or purple-black rice because only the surface of hulled rice contains pigments. These findings suggest that the outer portion contains various compounds other than starch and the inner portion contains relatively pure starch. Rice palatability and other characteristics can be improved through controlling the degree of milling using the biased distribution of chemical constituents within a rice kernel.

Protective effect of colored rice over white rice on Fenton reaction-based renal lipid peroxidation in rats.

Rice has been one of the most important grains. While polished white rice is favored, colored strains of rice, red, or black, have been maintained for religious purposes in Japan. We studied whether feeding of unpolished colored rice instead of white rice ameliorates oxidative renal tubular damage in rats induced by ferric nitrilotriacetate. Whereas renal lipid peroxidation was exacerbated in white rice-fed group in comparison with standard chow group, this exacerbation was not observed in red or black rice-fed groups. These changes were dependent on the proportion of colored rice to standard chow in the diet. Cyanidin 3-O-beta-D-glucoside was detectable neither in the serum nor kidney after one week of colored rice diet, but serum protocatechuic acid was significantly increased after black rice diet. There was a generalized decrease in the renal glutathione peroxidase activity in rice diet groups. Renal enzymatic activities of superoxide dismutase, glutathione S-transferase and NAD(P)H quinone reductase were not associated with the levels of lipid peroxidation. However, renal catalase activity was significantly increased in black rice-fed groups. These may partly explain the antioxidative effect. Furthermore, colored strains of rice are rich in proteins. Thus, our data warrants further investigation of the antioxidative effect of colored rice.