Genetic loci regulating the concentrations of anthocyanins and proanthocyanidins in the pericarps of purple and red rice.

The pigmented flavonoids, anthocyanins and proanthocyanidins, have health promoting properties. Previous work determined that the genes Pb and Rc turn on and off the biosynthesis of anthocyanins (purple) and proanthocyanidins (red), respectively. Not yet known is how the concentrations of these pigmented flavonoids are regulated in grain pericarps. Quantitative trait locus (QTL) analysis in a population of rice (Oryza sativa L.) F5 recombinant inbred lines from white pericarp "IR36ae" x red+purple pericarp "242" revealed three QTLs associated with grain concentrations of anthocyanins (TAC) or proanthocyanidins (PA). Both TAC and PA independently mapped to a 1.5 Mb QTL region on chromosome 3 between RM3400 (at 15.8 Mb) and RM15123 (17.3 Mb), named qPR3. Across 2 years, qPR3 explained 36.3% of variance in TAC and 35.8% in PA variance not attributable to Pb or Rc. The qPR3 region encompasses Kala3, a MYB transcription factor previously known to regulate purple grain characteristics. Study of PbPbRcrc progeny showed that TAC of RcRc near isogenic lines (NILs) was 2.1-4.5x that of rcrc. Similarly, study of PbPbRcRc NILs, which had 70% higher PA than pbpbRcRc NILs, revealed a mutual enhancement, not a trade-off between these compounds that share precursors. This suggests that Pb and Rc upregulate genes in a shared pathway as they activate TAC and PA synthesis, respectively. This study provides molecular markers for facilitating marker-assisted selection of qPR3, qPR5, and qPR7 to enhance grain concentrations of pigmented flavonoids and documented that stacking Rc and Pb genes further increases both flavonoid compounds.

Effects of Differences in Resistant Starch Content of Rice on Intestinal Microbial Composition.

The aim of this study was to evaluate the effects of resistant starch (RS) and fat levels on the gut microbiome in C57BL/6 mice. Three levels of RS from three varieties of rice were the major source of carbohydrates and fat levels were low (10%) and high (39%). We confirmed that RS decreased the Firmicutes to Bacteroidetes ratio, increased SCFA production by higher Bacteroidaceae and S24-7 abundance, and enriched predicted gene families of glycosidases and functional pathways associated with carbohydrate and glycan metabolism. We also found correlations between microbial taxa and tissue gene expression related to carbohydrate and lipid metabolism. Moreover, increasing RS levels resulted in a molecular ecological network with enhanced modularity and interspecific synergy, which is less sensitive to high fat intervention. Overall, RS as low as 0.44% from cooked rice can modulate gut microbiome in mice, which correlated to a protective effect against deleterious effects of an obesogenic diet.

Antioxidant Profile and Biosafety of White Truffle Mycelial Products Obtained by Solid-State Fermentation.

Solid-state fermentation may produce therapeutic compounds with higher biomass or better product characteristics than those generated by submerged fermentation. The objectives of this study were to analyze the antioxidant activities and biosafety of products obtained by white truffle (Tuber magnatum) solid-state fermentation in media with different ratios of soybean and red adlay. High levels of antioxidant components and high antioxidant activities such as DPPH radical scavenging, ferrous ion chelation, and reducing power were measured in 20 mg/mL water and ethanol extracts of the white truffle fermentation products. When the solid-state fermentation medium contained soybean and red adlay in a 1:3 ratio (S1A3), the fermentation product had more uniform antioxidant compositions and activities by principal component analysis (PCA). In addition, a 200 ppm water extract of the mycelial fermentation product was able to protect zebrafish embryos from oxidative stress induced by 5 mM hydrogen peroxide. Sprague-Dawley rats were fed the mycelial fermentation product for 90 consecutive days, revealing a no-observed-adverse-effect level (NOAEL) of 3000 mg/kg BW/day. Therefore, mycelial products obtained by white truffle solid-state fermentation can be used instead of expensive fruiting bodies as a good source of antioxidant ingredients.

High Resistant Starch Rice: Variation in Starch Related SNPs, and Functional, and Sensory Properties.

Human diets containing greater resistant starch (RS) are associated with superior glycemic control. Although high amylose rice has higher RS (29 g/kg to 44 g/kg) than lower amylose content varieties, sensory and processing properties associated with RS have not been evaluated. This study used variants of Waxy and starch synthase II a (SSIIa) genes to divide high amylose (256 g/kg to 284 g/kg) varieties into three haplotypes to examine their effects on RS, RVA parameters, and 14 cooked rice texture properties. RVA characteristics were influenced by both genes with peak and hotpaste viscosity differentiating the three haplotypes. Setback from hotpaste viscosity was the only RVA parameter correlated with RS content across three haplotypes (r = -0.76 to -0.93). Cooked rice texture attributes were impacted more by Waxy than by SSIIa with initial starch coating, roughness, and intact particles differentiating the three haplotypes. Pairwise correlation (r = 0.46) and PCA analyses suggested that roughness was the only texture attribute associated with RS content; while protein content influenced roughness (r = 0.49) and stickiness between grains (r = 0.45). In conclusion, variation exists among genetic haplotypes with high RS for sensory traits that will appeal to diverse consumers across the globe with limited concern for negatively affecting grain processing quality.

Effects of Rice with Different Amounts of Resistant Starch on Mice Fed a High-Fat Diet: Attenuation of Adipose Weight Gain.

Increasing the amount of resistant starch (RS) in the diet may confer protective effects against chronic diseases. Rice, a good dietary source of carbohydrates, also contains RS. However, it remains unclear if RS at the amount consumed in cooked rice has a health benefit. To address the question, we examined the effects of cooked rice containing different levels of RS in a diet-induced obesity rodent model. Rice containing RS as low as 1.07% attenuated adipose weight and adipocyte size gain, induced by a moderately high-fat (HF) diet, which correlated with lower leptin levels in plasma and adipose tissue. Rice with 8.61% RS increased fecal short-chain fatty acid levels, modulated HF-diet-induced adipose triacylglycerol metabolism and inflammation-related gene expression, and increased fecal triglyceride excretion. Hence, including rice with RS level at ≥1.07% may attenuate risks associated with the consumption of a moderately HF diet.

Association Analysis of Three Diverse Rice (Oryza sativa L.) Germplasm Collections for Loci Regulating Grain Quality Traits.

Rice ( L.) end-use cooking quality is vital for producers and billions of consumers worldwide. Grain quality is a complex trait with interacting genetic and environmental factors. Deciphering the complex genetic architecture associated with grain quality provides essential information for improved breeding strategies to enhance desirable traits that are stable across variable climatic and environmental conditions. In this study, genome-wide association (GWA) analysis of three rice diversity panels, the USDA rice core subset (1364 accessions), the minicore (MC) (173 accessions after removing non-), and the high density rice array-MC (HDMC) (383 accessions), with simple sequence repeats, single nucleotide polymorphic markers, or both, revealed large- and small-effect loci associated with known genes and previously uncharacterized genomic regions. Clustering of the significant regions in the GWA results suggests that multiple grain quality traits are inherited together. The 11 novel candidate loci for grain quality traits and the seven candidates for grain chalk identified are involved in the starch biosynthesis pathway. This study highlights the intricate pleiotropic relationships that exist in complex genotype-phenotypic associations and gives a greater insight into effective breeding strategies for grain quality improvement.

Hydrolytic rancidity and its association with phenolics in rice bran.

Whole grain rice, which has the bran layer intact, contains more nutrients and health beneficial compounds than its milled rice equivalent. Its consumption is associated with a reduction in the risk of developing several chronic diseases. However, the bran contains non-starch lipids deposited along with the lipid degrading enzymes, lipase and lipoxygenase, resulting in a relatively short shelf life for whole grain rice. We studied the genotypic diversity of lipase induced hydrolytic rancidity (HR) level in the bran of 134 diverse genotypes and found more than a 15-fold variation. Among the genotypes, those with red or brown bran had lower HR than the purple, light brown and white brans. Total phenolic content and anthocyanins were negatively correlated with the HR in purple brans suggesting their inhibitory effect on lipase during bran storage. In conclusion, low HR genotypes could be used as breeding materials to improve the storage stability of whole grain rice.

In Vitro Fermentation Patterns of Rice Bran Components by Human Gut Microbiota.

Whole grain rice is a rich source of fiber, nutrients, and phytochemicals that may promote gastrointestinal health, but such beneficial components are typically removed with the bran during polishing. Soluble feruloylated arabinoxylan oligosaccharides (FAXO) and polyphenols (RBPP) isolated from rice bran are hypothesized to have positive impacts on human gut microbiota through a prebiotic function. Using an in vitro human fecal fermentation bioassay, FAXO and RBPP treatments were assessed for short-chain fatty acids (SCFA) production patterns and by evaluating their impacts on the phylogentic composition of human gut microbiota by 16S rRNA gene sequencing. Fresh fecal samples collected from healthy adults (n = 10, 5 males, 5 females) were diluted with anaerobic medium. Each sample received five treatments: CTRL (no substrates), FOS (fructooligosaccharides), FAXO, RBPP, and MIX (FAXO with RBPP). Samples were incubated at 37 °C and an aliquot was withdrawn at 0, 4, 8, 12, and 24 h Results showed that SCFA production was significantly increased with FAXO and was comparable to fermentation with FOS, a well-established prebiotic. RBPP did not increase SCFA productions, and no significant differences in total SCFA production were observed between FAXO and MIX, indicating that RBPP does not modify FAXO fermentation. Changes in microbiota population were found in FAXO treatment, especially in Bacteroides, Prevotella, and Dorea populations, indicating that FAXO might modulate microbiota profiles. RBPP and MIX increased Faecalibacterium, specifically F. prausnitzii. Combined FAXO and RBPP fermentation increased abundance of butyrogenic bacteria, Coprococcus and Roseburia, suggesting some interactive activity. Results from this study support the potential for FAXO and RBPP from rice bran to promote colon health through a prebiotic function.

Resistant starch: Variation among high amylose rice varieties and its relationship with apparent amylose content, pasting properties and cooking methods.

Resistant starch (RS), which is not hydrolyzed in the small intestine, has proposed health benefits. We evaluated 40 high amylose rice varieties for RS content in cooked rice and a 1.9-fold difference was found. Some varieties had more than two-fold greater RS content than a US long-grain intermediate-amylose rice. The high amylose varieties were grouped into four classes according to paste viscosity and gelatinization temperature based on genetic variants of the Waxy and Starch Synthase IIa genes, respectively. RS content was not different between the four paste viscosity-gelatinization temperature classes. Multiple linear regression analysis showed that apparent amylose content and pasting temperature were strong predictors of RS within each class. Two cooking methods, fixed water-to-rice ratio/time and in excess-water/minimum-cook-time, were compared using six rice varieties that were extremes in RS in each of the genetic variant classes, no difference in RS content due to cooking method was observed.

Antidiabetic Potential of Purple and Red Rice (Oryza sativa L.) Bran Extracts.

Pigmented rice contains anthocyanins and proanthocyanidins that are concentrated in the bran layer. In this study, we determined the phenolic, flavonoid, anthocyanin, and proanthocyanidin content of five rice bran (1 brown, 2 red, and 2 purple) extracts. Each bran extract was evaluated for inhibitory effects on α-amylase and α-glucosidase activity, two key glucosidases required for starch digestion in humans. All purple and red bran extracts inhibited α-glucosidase activity, however only the red rice bran extracts inhibited α-amylase activity. Additionally, each bran extract was examined for their ability to stimulate glucose uptake in 3T3-L1 adipocytes, a key function in glucose homeostasis. Basal glucose uptake was increased between 2.3- and 2.7-fold by exposure to the red bran extracts, and between 1.9- and 3.1-fold by exposure to the purple bran extracts. In red rice bran, the highest enzyme inhibition and glucose uptake was observed with a proanthocyanidin-enriched fraction. Both IITA red bran and IAC purple bran increased expression of GLUT1 and GLUT4 mRNA, and genes encoding insulin-signaling pathway proteins.

Bran data of total flavonoid and total phenolic contents, oxygen radical absorbance capacity, and profiles of proanthocyanidins and whole grain physical traits of 32 red and purple rice varieties.

Phytochemicals in red and purple bran rice have potential health benefit to humans. We determined the phytochemicals in brans of 32 red and purple global rice varieties. The description of the origin and physical traits of the whole grain (color, length, width, thickness and 100-kernel weight) of this germplasm collection are provided along with data of total flavonoid and total phenolic contents, oxygen radical absorbance capacity and total proanthocyanidin contents. The contents and proportions of individual oligomers, from degree of polymerization of monomers to 14-mers, and polymers in bran of these 32 rice varieties are presented (DOI: http://dx.doi.org/10.1016/j.foodchem.2016.04.004) [1].

Concentrations of oligomers and polymers of proanthocyanidins in red and purple rice bran and their relationships to total phenolics, flavonoids, antioxidant capacity and whole grain color.

Proanthocyanidins, a flavonoids subgroup, are proposed to have chronic disease modulation properties. With the eventual goal of enhancing rice phytonutrient concentrations, we investigated the genotypic variation of the concentrations of individual oligomers and polymers of proanthocyanidins in red and purple rice brans. A 4.3-fold variation in total proanthocyanidins (sum of oligomers and polymers) in the extractable fraction was found and the concentration was highly correlated with total phenolics, total flavonoids and antiradical capacity. Variation in the proportion of oligomers and polymers existed, with monomers to trimers, 4-6mers, 7-10mers and polymers accounting for 7, 18, 26.5 and 48.7%, respectively, of the total. The redness value a(∗) of whole grain rice measured in CIE L(∗)a(∗)b(∗) color space was negatively and positively correlated with extractable and non-extractable proanthocyanidins, respectively. The variation found indicates it is possible to select rice with bran containing high levels of total proanthocyanidins and specific degree of polymerization profiles.

Effects of hydrothermal processes on antioxidants in brown, purple and red bran whole grain rice (Oryza sativa L.).

The impacts of parboiling and wet-cooking, alone and in combination, on the concentrations of lipophilic antioxidants (vitamin E and γ-oryzanol), soluble (including proanthocyanidins and anthocyanins) and cell wall-bound phenolics, and antioxidant capacities in whole grain rice from six cultivars having different bran colours were investigated. Parboiling rough and brown rice increased the concentrations of lipophilic antioxidants in whole grain rice but decreased the concentrations of total phenolics and antioxidant capacities found in the soluble fraction. After hydrothermal processing of purple bran rice, the retention of extractable anthocyanins was low, but was high for simple phenolics. For proanthocyanidins found in red bran rice, the extractable oligomers with a degree of polymerization (DP) less than 4, increased up to 6-fold; while for oligomers with DP⩾4 and polymers, there was a significant decrease that was positively correlated with the DP and the temperature of the processing methods. The presence of hulls helped to retain water-soluble antioxidants during parboiling.

Rice varietal differences in bioactive bran components for inhibition of colorectal cancer cell growth.

Rice bran chemical profiles differ across rice varieties and have not yet been analysed for differential chemopreventive bioactivity. A diverse panel of seven rice bran varieties was analysed for growth inhibition of human colorectal cancer (CRC) cells. Inhibition varied from 0% to 99%, depending on the variety of bran used. Across varieties, total lipid content ranged 5-16%, individual fatty acids had 1.4- to 1.9-fold differences, vitamin E isoforms (α-, γ-, δ-tocotrienols, and tocopherols) showed 1.3- to 15.2-fold differences, and differences in γ-oryzanol and total phenolics ranged between 100-275ng/mg and 57-146ngGAE/mg, respectively. Spearman correlation analysis was used to identify bioactive compounds implicated in CRC cell growth inhibitory activity. Total phenolics and γ-tocotrienol were positively correlated with reduced CRC cell growth (p<0.05). Stoichiometric variation in rice bran components and differential effects on CRC viability merit further evaluation elucidate their role in dietary CRC chemoprevention.

Identification of an abundant 56 kDa protein implicated in food allergy as granule-bound starch synthase.

Rice, the staple food of south and east Asian counties, is considered to be hypoallergenic. However, several clinical studies have documented rice-induced allergy in sensitive patients. Rice proteins with molecular weights of 14-16, 26, 33, and 56 kDa have been identified as allergens. Recently, it was documented that the 56 kDa rice allergen was responsible for rice-induced anaphylaxis. The 14-16 kDa allergens have been identified as α-amylase inhibitors; the 26 kDa protein has been identified as α-globulin; and the 33 kDa protein has been identified as glyoxalase I. However, the identity of the 56 kDa rice allergen has not yet been determined. In this study, we demonstrate that serum from patients allergic to maize shows IgE binding to a 56 kDa protein that was present in both maize and rice but not in the oil seeds soybean and peanut. The 56 kDa IgE-binding protein was abundant in the rice endosperm. We have purified this protein from rice endosperm and demonstrated its reactivity to IgE antibodies from the serum of maize-allergic patients. The purified protein was subjected to matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry analysis, resulting in identification of this rice allergen as granule-bound starch synthase, a product of the Waxy gene. Immunoblot analysis using protein extracts from a waxy mutant of rice revealed the absence of the 56 kDa IgE-binding protein. Our results demonstrate that the 56 kDa rice allergen is granule-bound starch synthase and raise the possibility of using waxy mutants of rice as a potential source of the hypoallergenic diet for patients sensitized to the 56 kDa rice allergen.

Growth-inhibitory effects of pigmented rice bran extracts and three red bran fractions against human cancer cells: relationships with composition and antioxidative activities.

We determined the phenolic, anthocyanin, and proanthocyanidin content of three brown, purple, and red rice brans isolated from different rice varieties using HPLC-PDA with the aid of 27 standards of known structure and matching unknown peaks to a spectral library of known compounds. Antioxidative capacities were determined by DPPH and ORAC and cell-inhibiting effects using an MTT assay. Based on the calculated IC(50) values, the light-brown bran had no effect, the purple bran exhibited a minor effect on leukemia and cervical cancer cells, and the red bran exhibited strong inhibitory effects on leukemia, cervical, and stomach cancer cells. High concentrations of protocatechuic acid and anthocyanins in purple bran and proanthocyanidins in red bran were identified. The red bran was further fractionated on a Sephadex column. Fraction 3 rich in proanthocyanidin oligomers and polymers had the greatest activity. Red bran has the potential to serve as a functional food supplement for human consumption.

Phytochemicals and antioxidant capacities in rice brans of different color.

Rice bran, a byproduct of the rice milling process, contains most of the phytochemicals. This study aimed at determining the concentrations of lipophilic, solvent-extractable (free), and cell wall-bound (bound) phytochemicals and their antioxidant capacities from brans of white, light brown, brown, purple, and red colors, and broccoli and blueberry for comparison. The concentrations of lipophilic antioxidants of vitamin E (tocopherol and tocotrienols) and γ-oryzanols were 319.67 to 443.73 and 3861.93 to 5911.12 µg/g bran dry weight (DW), respectively, and were not associated with bran color. The total phenolic, total flavonoid, and antioxidant capacities of ORAC (oxygen radical absorbance capacity), DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging, and iron-chelating in the free fraction were correlated with the intensity of bran color, while variations of these in the bound fraction were less than those in the free fraction among brans. Compounds in the bound fraction had higher antioxidant capacity of ORAC than DPPH, relative to those in the free fraction. The bound fraction of light-color brans contributed as much to its total ORAC as the free fraction. Total proanthocyanidin concentration was the highest in red rice bran, while total anthocyanin was highest in purple brans. The predominant anthocyanin was cyanidin-3-glucoside. Red and purple brans had several fold higher total phenolics and flavonoids as well as ORAC and DPPH, from both free and bound fractions, than freeze-dried blueberry and broccoli. These results indicate that rice brans are natural sources of hydrophilic and lipophilic phytochemicals for use in quality control of various food systems as well as for nutraceutical and functional food application.

Metabolomic and functional genomic analyses reveal varietal differences in bioactive compounds of cooked rice.

Emerging evidence supports that cooked rice (Oryza sativa L.) contains metabolites with biomedical activities, yet little is known about the genetic diversity that is responsible for metabolite variation and differences in health traits. Metabolites from ten diverse varieties of cooked rice were detected using ultra performance liquid chromatography coupled to mass spectrometry. A total of 3,097 compounds were detected, of which 25% differed among the ten varieties. Multivariate analyses of the metabolite profiles showed that the chemical diversity among the varieties cluster according to their defined subspecies classifications: indica, japonica, and aus. Metabolite-specific genetic diversity in rice was investigated by analyzing a collection of single nucleotide polymorphisms (SNPs) in genes from biochemical pathways of nutritional importance. Two classes of bioactive compounds, phenolics and vitamin E, contained nonsynonymous SNPs and SNPs in the 5' and 3' untranslated regions for genes in their biosynthesis pathways. Total phenolics and tocopherol concentrations were determined to examine the effect of the genetic diversity among the ten varieties. Per gram of cooked rice, total phenolics ranged from 113.7 to 392.6 µg (gallic acid equivalents), and total tocopherols ranged between 7.2 and 20.9 µg. The variation in the cooked rice metabolome and quantities of bioactive components supports that the SNP-based genetic diversity influenced nutritional components in rice, and that this approach may guide rice improvement strategies for plant and human health.