Changes of structure and functional properties of rice protein in the fresh edible rice during the seed development

It has been reported that fresh edible rice has more bioactive compounds and its protein is easier to digest and has lower hypoallergenic than mature rice. In this paper, the changes in structure and functional properties of proteins at five different stages, including early milky stage(EMS), middle milky stage(MMS), late milky stage(LMS), waxy ripe stage(WS)and ripening stage(RS), during the seed development were investigated. It was found that with the seed developing, the molecular weight of fresh rice protein gradually become larger while the secondary structure changed from the highest content of disordered structure at MMS to the highest content of ordered structure at RS, which affect the surface hydrophobicity and then the functional properties of proteins, including foaming properties, emulsifying properties and oil holding capacity. Fresh rice protein at MMS has the strongest surface hydrophobicity while fresh edible rice protein at RS has the strongest oil holding capability. The results of our study can provide a theoretical basis for the application of fresh rice protein in the food industry and help to develop new fresh edible rice food.

Comparative Study of the Protein Contents of Local and Imported Rice Consumed in Senegal

Protein is essential for the growth and maintenance of the body. They play a crucial role in different biological processes. This study focuses on comparing the protein contents of local rice grown in the Senegal River valley and rice imported from Asia. The objective is to evaluate the importance of the protein nutritional value of local rice compared to imported rice. Protein contents were determined using the Kjeldahl method. The results of the protein assays show that local rice varieties such as Sahel 108 and Sahel 134 grown in the Thilène basins had protein percentages comparable to those of imported rice. The protein percentages were 15.19% ± 0.91% for the Sahel 108 variety and 16.62% ± 0.01% for the Sahel 134 variety compared to 15.8% ± 0.01% on average for imported rice. Thus from the point of view of protein content, local rice has a nutritional value identical to that of imported rice which it can validly replace. It is important in Senegal to encourage local production, which would make it possible to reduce imports on the one hand and to make quality rice available to the local Senegalese market on the other. Sahel varieties with high protein contents deserve large-scale development to meet the country’s protein needs.

PREPARATION AND PROPERTIES OF WHEAT GLUTEN/RICE PROTEIN COMPOSITES PLASTICIZED WITH GLYCEROL

Environment friendly thermosetting composites were prepared by blending wheat gluten （WG） and rice protein （RP） at different weight ratios with glycerol as plasticizer followed by compression molding the mixture at 120℃ to crosslink the proteins. Reducing agent of sodium bisulfate and sodium sulfite and crosslinking agent formaldehyde were used to adjust the properties of the composites. Morphology, moisture absorption and tensile properties were evaluated. The results showed that formaldehyde could increase tensile strength of the composites without significant influence on Young＇s modulus and elongation at break. On the other hand, reducing agents could improve tensile strength and extensibility simultaneously, which was much marked at WG/RP ratios from 7/3 to 3/7.

Investigating the effects of ion strength on amyloid fibril formation of rice proteins

Protein fibrillation is recognized as an attractive strategy to broaden and improve the function of food proteins.This research investigated the influence of different NaCl concentrations(0-210 mM)on the thermal aggregation behavior of rice proteins(RP)at pH 2.0.Under lower(<100 mM)and higher(>100 mM)NaCl concentrations,RP formed aggregates with different sizes and structures.At lower concentrations,the zeta-potential of RP increased from 14.3 mV to 22.8 mV during incubation,indicating that the electrostatic repulsion of RP was enhanced and numerous positively charged building blocks were generated.The circular dichroism(CD)results indicated that the native structure of RP was destroyed after heating at 90℃ for 24 h,and a secondary structure dominated byβ-sheets(more than 50%)was formed.Proteins formed long fibrils after 24 h heating,as observed by atomic force microscope(AFM).However,at higher NaCl concentrations,the hydrophobic interaction was the dominant force due to electrostatic shielding.The protein aggregated randomly to form amorphous aggregates with particle sizes over 600 nm.This study will provide a new insight of RP-based fibrils in food application.

Modeling Rice rgMT as a Plant Metallothionein-Like Protein by the Distance Geometry and Homology Methods

Rice metallothionein-like protein (rgMT) shows characteristics of a three-section pattern composed of two highly conserved cysteine rich (CR) domains in the terminals and a spacer without cysteine (cys) residues in the center of the molecule. In this paper, the two CR domains and the spacer region were modeled by the distance geometry and homology methods separately. For the CR domains, twenty random models were generated for each cys combination based on the constraint conditions of CXC (C represents cys, X represents any amino acid other than cys), and CXXC motifs and a metal-sulfur chelating cluster. Four models for the N-terminal and two for C-terminal CR domain containing metal chelating structures formed by different combinations of cys were selected from 900 possible conformations. The GOR method was used to predict the secondary structure of the spacer region and its model was built by the homology method. After three parts of the protein were modeled, they were connected to form a three-dimensional structure model of rgMT. The whole conformation showed that rgMT could form two independent metal-sulfur chelating structures connected by a spacer peptide, without a structural or energy barrier for them to form two independent metal-chelating clusters just as mammalian metallothionein (MT) proteins. As all plant metallothionein-like (MT-L) proteins have the same primary structural characteristic, two CR domains connected by a spacer region, and many have the same cys arrangement pattern as rgMT, the three-dimensional structure model of rgMT will provide an important reference for the structural study of other plant MT-L proteins.

Rice Storage Proteins:Focus on Composition,Distribution,Genetic Improvement and Effects on Rice Quality

Rice storage proteins(RSPs)are plant proteins with high nutritional quality.As the second largest type of storage substance in rice,it is the main source of protein intake for people who consume rice as a staple food.The content and type of RSPs affect the appearance,processing quality and eating quality of rice.These effects involve the distribution of RSPs in rice grains as well as the interactions of RSPs with other components such as starch in rice grains.In the past two decades,some progress has been made in the genetic improvement of RSPs.However,the determination mechanism of protein content and composition in rice is still unclear,and the mechanism of the effect of RSPs on rice quality has not been elucidated.In this review,the composition,biosynthesis and distribution of RSPs,and quantitative trait loci mapping and cloning of RSP genes are summarized,the research progress of the influence of RSPs and their components on rice quality are reviewed,and the research directions in the future are proposed.

Charactering protein fraction concentrations as influenced by nitrogen application in low-glutelin rice cultivars

To optimize both grain yield and quality of low-glutelin rice cultivars under N-fertilizer strategies, two-year field experiments involving three low-glutelin rice cultivars（W1240, W1721, W025） and an ordinary rice cultivar（H9405） with five N treatments were carried out to determine the effects of N application rate and genotype on protein fractions contents and Glutelin/Prolamin ratio（Glu/Pro）. The difference of protein fraction concentrations affected by N application rate existed in genotypes. Ordinary rice cultivar had a larger increase in glutlein concentration affected by N application rate than low-glutelin rice cultivars did. Glutelin in H9405 had a increase of 30.6 and 41.0% under the N4 treatment（360 kg N ha^（–1）） when compared with N0 treatment（no fertilizer N） in 2010 and 2011 respectively, while all the low-glutelin rice cultivars showed relatively smaller increases for two years. Variance analysis showed no significant effect of N application rate on glutelin in W1240 and W025 while the effects on albumin, globulin and prolamin were significant in low-glutelin rice. What＇s more, N treatment had no significant i nfluence on Glu/Pro ratios in low-glutelin rice cultivars while a significant increase in Glu/Pro ratio was observed in ordinary rice cultivar. So low-gultelin rice cultivars showed a different pattern from ordinary rice cultivars when influenced by N application rate.

Western blot detection of PMI protein in transgenic rice

Phosphomannose isomerase （PMI） encoding gene manA is a desirable selective marker in transgenic research. Under- standing of its expression patterns in transgenic plant and establishing highly sensitive detection method based on immunoassay have great impacts on the application of PMI. In this study, PMI-specific monoclonal antibodies were generated using recombinant protein as immunogen, and could be used in Western blot to detect as little as 0.5 ng His-tagged PMI protein or rice expressed PMI protein in sample accounted for 0.4% of single rice grain （about 0.08 mg）. PMI protein driven by CaMV-35S promoter was detected in dozens of tested tissues, including root, stem, leaf, panicle, and seed at all developmental stages during rice growing, and PMI protein accounted for about 0.036% of total protein in the leaves at seedling stage. The established method potentially can be used to monitor PMI protein in rice grains.

Effects of Uniconazole on Nitrogen Metabolism and Grain Protein Content of Rice

The effects of uniconazole by soaking seeds and spraying leaves at booting stage with different concentrations （0, 20 and 40 mg/kg） on the nitrogen metabolism of flag leaf and grains after flowering, and rice grain protein content and yield were studied with hybrid rice combination Shanyou 63. Under uniconazole treatment, the soluble protein content in flag leaf was increased in early and middle period of grain filling, but this content was nearly the same as or even lower than that of control at maturity; Glutamine synthetase activity in superior and inferior grains and non-protein nitrogen content in superior grains at early stage of grain development were promoted, and moreover, the transforming speed from non-protein nitrogen to protein nitrogen was accelerated; Non-protein nitrogen content was lower than that of control at maturity, but protein nitrogen content at each stage was higher than those of control; Protein nitrogen content in superior and inferior grains and protein nitrogen absolutely accumulative content in a grain both were enhanced and protein content and yield in rice grain were raised. The application of uniconazole by soaking seeds and spraying leaves raised crude protein content by an average of 7.2% and 8.3%, and protein yield by an average of 13.1% and 13.4%, respectively.

Transformation of Sucrose to Starch and Protein in Rice Leaves and Grains under Two Establishment Methods

Six rice varieties, PR120, PR116, Feng Ai Zan, PR115, PAU201 and Punjab Mehak 1 were raised under aerobic and transplanting conditions to assess the effects of planting conditions on sucrose metabolising enzymes in relation to the transformation of free sugars to starch and protein in flag leaves and grains. Activities of sucrose synthase, sucrose phosphate synthase and acid invertase increased till flowering stage in leaves and mid-milky stage（14 d after flowering） in grains and thereafter declined in concomitant with the contents of reducing sugar. Under aerobic conditions, the activities of acid invertase and sucrose synthase（cleavage） significantly decreased in conjunction with the decrease in non-reducing sugars and starch content in all the varieties. Disruption of starch biosynthesis under the influence of aerobic conditions in both leaves and grains and the higher build up of sugars possibly resulted in their favoured utilization in nitrogen metabolism. Feng Ai Zan, PR115 and PR120 maintained higher levels of sucrose synthase enzymes in grains and leaves and contents of metabolites（amino acid, protein and non-reducing sugar） under aerobic conditions, while PR116, Punjab Mehak 1 and PAU201 performed better under transplanting conditions, thus showing their adaptation to environmental stress. Yield gap between aerobic and transplanting rice is attributed primarily to the difference in sink activity and strength. Overall, it appear that up-regulation of sucrose synthase（synthesis） and sucrose phosphate synthase under aerobic conditions might be responsible in enhancing growth and productivity of rice varieties.

Effects of Different Nitrogen Fertilizer Levels and Native Soil Properties on Rice Grain Fe, Zn and Protein Contents

Deposition of protein and metal ions （Fe, Zn） in rice grains is a complex polygenic trait showing considerable environmental effect. To analyze the effect of nitrogen application levels and native soil properties on rice grain protein, iron （Fe） and zinc （Zn） contents, 32 rice genotypes were grown at three different locations each under 80 and 120 kg/hm2 nitrogen fertilizer applications. In treatments with nitrogen fertilizer application, the brown rice grain protein content （GPC） increased significantly （1.1% to 7.0%） under higher nitrogen fertilizer application （120 kg/hm2） whereas grain Fe/Zn contents showed non-significant effect of nitrogen application level, thus suggesting that the rate of uptake and translocation of macro-elements does not influence the uptake and translocation of micro-elements. The pH, organic matter content and inherent Fe/Zn levels of native soil showed significant effects on grain Fe and Zn contents of all the rice genotypes. Grain Zn content of almost all the tested rice genotypes was found to increase at Location III having loamy soil texture, neutral pH value （pH 6.83） and higher organic matter content than the other two locations （Locations I and II）, indicating significant influence of native soil properties on brown rice grain Zn content while grain Fe content showed significant genotype × environment interaction effect. Genotypic difference was found to be the most significant factor to affect grain Fe/Zn contents in all the tested rice genotypes, indicating that although native soil properties influence phyto-availability of micronutrients and consequently influencing absorption, translocation and grain deposition of Fe/Zn ions, yet genetic makeup of a plant determines its response to varied soil conditions and other external factors. Two indica rice genotypes R-RF-31 （27.62 μg/g grain Zn content and 7.80% GPC） and R1033-968-2-1 （30.05 μg/g grain Zn content and 8.47% GPC） were identified as high grain Zn and moderate GPC rice genotypes. These results indicate that soil property and organic matter content increase the availability of Fe and Zn in rhizosphere, which in turn enhances the uptake, translocation and redistribution of Fe/Zn into rice grains.

Genetic Diversity of Seed Storage Proteins in Different Ecotype Varieties of japonica Rice and Its Application

One hundred and fifteen varieties （including cultivars and lines） with different ecotypes in japonica rice （Oryza sativa L.） were analyzed for endosperm storage proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） to estimate their genetic diversity for the purpose of genetic improving and variety identification. Nineteen types of profile were identified according to 1） presence/absence of 65 kDa bands, 2） staining intensity of 70, 60, 57, 37-39, 22-23, 13 and 10 kDa bands, 3） migration velocity of 35 kDa （α-4） and 4） band number at 57 kDa location. An unweighted-pair group average method with arithmetic mean （UPGMA） dendregram based on the cluster analysis of genetic similarity of the protein bands showed a small genetic variation among the tested materials, with the similarity coefficients varying between 0.75 and 1.00. Three distinct groups were identified from the cluster analysis of the rice varieties studied at the similarity coefficient level of 0.894. The first group included eight varieties with high amylose content, the second group contained fifteen varieties with high protein content, and the third group had the remaining ninety-two varieties, which accounted for 80% of the total materials. Clear relationship between ecotypes distinguished by maturity and groups revealed by cluster analysis was not found in this study. Only the group of high amylose linked with medium-maturity medium japonica ecotype. The bands of 70 kDa and 65 kDa can be used as protein markers to identify F1 seed purity of japonica hybrid rice Liuyanyou 422.

Effects and Mechanisms of P and K Nutrients on Yield and Protein Content of Fodder Rice

Effects and mechanisms of P and K nutrients on yield and protein content of Weiyou 56, a fodder hybrid rice combination,were studied through pot experiment and biochemical analysis. The results showed that the increase of P and K nutrients enhanced the activities of PEP carboxylase (PEPC), glutamine synthase (GS) and sucrose phosphate synthase (SPS) in leaves, sucrose synthase (SS), ADP glucose pyrophosphorylase (ADPGP) and GS in grains, and the chlorophyll content in leaves, soluble sugar and starch content in grains, protein N and total N content in leaves and grains. Howerer, they decreased soluble sugar content in leaves and led to an increase of protein content in brown rice, biomass, grain yield and harvest index. Excessive P nutrients slightly reduced SPS and ADPG activity in leaves and grains respectively.

Impacts of Environmental Factors on Degradation of CrylAb Insecticidal Protein in Leaf-Blade Powders of Transgenic Bt Rice

The determination of the environmental fate of Bt insecticidal protein released by Bt rice plants in paddy soils is a key issue in its ecological risk assessment. In this study, the impacts of soil water content, pH, and temperature on the degradation of CrylAb protein expressed in the leaves of Bt rice KMD2 were studied in the laboratory. Three types of paddy soils were used, i.e., blue clayey paddy soil, pale paddy soil on quaternary red soil, and marine-fluvigenic yellow loamy paddy soil. Ground powders of KMD2 leaf blades were mixed with each type of soil, and degradation dynamics of Cry lAb were measured using enzyme-linked immunosorbent assay （ELISA）. The degradation rate of CrylAb was high at the early experimental stage, but slowed down steadily at middle and later stages, which could be described by exponential equations, with the half-life period of degradation determined as 1.8-4.0 d. The soil water content, pH, and temperature could affect the degradation of CrylAb, but the effects of soil pH and temperature were relatively greater. In general, CrylAb degradations were slower under lower soil pH and temperature conditions, especially for marine-fluvigenic yellow loamy paddy soil.

Differences in the Content of Protein and Essential Amino Acids between Different Rice Varieties

Evaluation of the protein content is a major indicator of the nutritional quality of rice,and the protein quality of rice is the best among cereal crops. Essential amino acids play an irreplaceable role in human growth,development and health care. Essential amino acid is a key ingredient to measure the nutritional value of rice. Using the experimental rice processed from the rice variety " Yuzhenxiang" sprayed with plant nutrients( patent number: ZL201110103910. 9),namely high essential amino acid nutritional rice,combined with five kinds of high quality rice imported by COFCO and homegrown " Wuchang rice",we send the samples of the seven kinds of rice to Hunan Food Testing Center,and adopt HPLC method to test the content of protein and eight kinds of essential amino acids. Three bags of rice are randomly selected for each kind of rice,and each bag is a replication. The test results show that there are highly significant differences in the content of essential amino acids between different kinds of rice( F = 246. 29**,P =5 ×10- 71),and there are also highly significant differences in the content between different kinds of essential amino acids( F = 3937. 09**,P = 4 × 10- 146). The test results of protein content indicate that there are highly significant differences in the content of protein between different kinds of rice( F = 3937. 0973. 29**,P =5. 81 ×10- 11),and the test results of lysine content show that there are highly significant differences in the content of lysine between different kinds of rice( F =3937. 0973. 29**,P =5. 81 ×10- 11).

Genetic Analysis and Improvement Strategies of Protein Content in Rice Seeds

Enhancing the nutritional quality of food crops is a way to improve human nutrition and health. Grain protein content （ GPC ） is an important nutritional quality trait for rice and many other crops. Therefore, it will be very significant to increase GPC and improve rice quality. This paper mainly reviewed the genetic studies GPC in rice, mapping and cloning of QTLs and genes associated with GPC. However, little is known about the molecular mechanism and gene function related to GPC in rice. Thus, here we put forward the genetic strategy by gene pyramiding or molecular marker assisted selection to improve GPC and nutritional quality in rice. This review could provide important information for genetic improvement of grain nutritional quality of rice.

Identification of a novel gain-of-function mutant allele,slr1-d5,of rice DELLA protein

Controling the height of crops plays a crucial role for their yields. The large scale utilization of semi-dwarf varieties has greatly improved crop yield, providing an effective support for world food security. In rice, a main food for over half of the world’s population, a number of dwarf loci have been identiifed. However, most of them are recessive, such as the ‘green revolution’ genesd1. To gain more beneifcial loci for rice breeding programs, exploring new mutations is needed, especialy the dominant loci which can be used broadly for hybrid breeding. Here, we isolated a novel dominant dwarf rice mutant, slr1-d5. Al of the internodes ofslr1-d5are reduced. We ifnd that the responsiveness ofslr1-d5to gibberelin （GA）, GA3, was signiifcantly reduced. Map-based cloning revealed that the dominant dwarifsm ofslr1-d5was caused by an amino acid substitution in the N-terminal TVHYNP domain of rice DELLA protein, SLR1, where the conserved amino acid Pro （P） was substituted to His （H）. Our ifndings not only further prove the pivotal role of TVHYNP motif in regulating SLR1 stability, but also provide a new dwarf source for improvement of rice germplasms.

Analysis of Male Sterility-Related Proteins of Young Panicle in Rice (Oryza sativa L.) by Two-Dimensional Electrophoresis

For searching out male sterility-related proteins (polypeptides) in rice(Oryza sativa L. ), we examined the difference of panicle protein (polypeptides) between hybrid rice( Wujin2A/R168, Wujin5A/R988) and their parents (male-sterile line Wujin2A, Wujin5A, and restorerline R168, R988) at the formation stage of pollen mother cell by two-dimensional electrophoresis(2-DE). The results revealed that the 2-DE polypeptide maps were similar among these experimentalmaterials. A small group of polypeptides were disappeared in 2-DE polypeptide maps of male-sterileline (Wu-jin2A, WujinSA) by comparing to restorer line (R168, R988) and the first filial (F, )generation (Wujin2A/R168, Wujin5A/R988). The isoelectric points of these polypeptides were pl5.8-6.5, molecular weight 42. 7 X 10~3-66. 2 X 10~3.

Characterization of Agro-diversity by Seed Storage Protein Electrophoresis:Focus on Rice Germplasm from Uttarakhand Himalaya,India

The characteristics of 48 rice varieties from Uttarakhand Himalaya, India were detected by morphological and biochemical markers. The grains of the selected rice varieties varied in their morphological （grain length, grain width and grain weight） and biochemical characters （sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE）. Based on the presence of 70, 65, 60, 57, 37-39, 22-23, 13 and 10 kDa protein bands in the 48 rice varieties, seven types of profiles were identified. An unweighted pair group average method with arithmetic mean （UPGMA） dendrogram based on cluster analysis of genetic similarity of the protein bands showed two distinct groups with 1%-78% similarity coefficients. The presence of characteristic bands in selected varieties is a useful parameter for identification of rice germplasm.

Cloning and Expression Analysis of a Mitogen-Activated Protein Kinase Gene OsMPK14 in Rice

Mitogen activated-protein kinases （MAPKs） are important components in signal transduction pathways responding to various biotic and abiotic stresses. An MAPK gene, OsMPK14 （GenBank Accession No. GQ265780） from rice （Oryza sativa L.）, was cloned by RT-PCR. The full-length cDNA of OsMPK14 consists of 1660 bp in size, containing an open reading frame of 1629 bp, which encodes a 542-amino-acid polypeptide and has a typical protein kinase domain and a phosphorylation activation motif TDY. Sequence alignment and analysis revealed that OsMPK14 was located on rice chromosome 5, and composed of nine exons and eight introns in the coding region. Semi-quantitative RT-PCR was performed to detect the expression patterns of OsMPK14 in rice shoots and roots under darkness, drought, high salinity, low temperature and abscisic acid treatments. The OsMPK14 mRNA was induced by abscisic acid, low temperature and high salinity, but weakly inhibited by drought. In addition, the expression of OsMPK14 was up-regulated in roots, but down-regulated in shoots by light. The results indicate that OsMPK14 could be implicated in diverse rice stimuli-responsive signaling cascades, and its expression might be regulated by multiple factors.