Germinated brown rice relieves hyperlipidemia by alleviating gut microbiota dysbiosis

Hyperlipidemia is a frequent metabolic disorder that is closely associated with diet. It is believed that brown rice, containing the outer bran layer and germ, is beneficial for the remission of hyperlipidemia. This study established a rat model of hyperlipidemia by feeding a high-fat diet. The hypolipidemic potential of germinated brown rice(Gbrown) and germinated black rice(a germinated black-pigmented brown rice, Gblack) were explored in the model rats, mainly in the aspects of blood lipids, lipases, apolipoproteins, and inflammation. The gut microbiota in hyperlipidemic rats receiving diverse dietary interventions was determined by 16S rDNA sequencing. The results showed that the intervention of Gbrown/Gblack alleviated the hyperlipidemia in rats, evidenced by decreased TC, TG, LDL-C, and apolipoprotein B, and increased HDL-C, HL, LPL, LCAT, and apolipoprotein A1. Gbrown/Gblack also weakened the inflammation in hyperlipidemia rats, evidenced by decreased TNF-α, IL-6, and ET-1. In addition, 16S rDNA sequencing revealed that the diet of Gbrown/Gblack elevated the abundance and diversity of gut microbiota in hyperlipidemia rats. At the phylum level, Gbrown/Gblack decreased Firmicutes, increased Bacteroidetes, and decreased the F/B ratio in hyperlipidemia rats. At the genus level, Gbrown/Gblack decreased Streptococcus and increased Ruminococcus and Allobaculum in hyperlipidemia rats. Some differential microbial genera relating to lipid metabolism were also determined, such as the Lachnospira and Ruminococcus in the Gblack group, and the Phascolarctobacterium, Dorea, Turicibacter, and Escherichia-Shigella in the Gbrown group. Notably, the beneficial effect of Gblack was stronger than Gbrown. To sum up, the dietary interventions of Gbrown/Gblack contributed to the remission of hyperlipidemia by alleviating the dysbiosis of gut microbiota.

Brown rice:a missing nutrient-rich health food

Brown rice(BR)is a traditional health food being rich in various active substances, which have effective preventive and therapeutic effects on many diseases. In this review, we systemically summarized the efficacy of BR on cardiovascular diseases(CVDs), hyperlipidemia, hypertension, diabetes, obesity, osteoporosis, neurodegenerative diseases, cancer, immune disorders, inflammation, anxiety, liver and stomach damage, etc. Underlying mechanisms have also been analyzed, which include regulating lipid synthesis and metabolism, improving insulin levels and increasing glucose metabolism, anti-oxidation, promoting the immune system, transcription and non-transcriptional regulation, blocking cell cycle, thereby improving disease symptoms and reducing its recurrence rate. Generally, BR has been almost disappeared in modern society, but it is promising to exploit the potential to become a functional food. Further research on the nutritional value, variety differences, storage problems, taste and molecular mechanism of BR will be conducive to help meticulously design clinical intervention trials and ultimately improve the treatment and prevention of various diseases.

Gapless Genome Assembly of ZH8015 and Preliminary Multi-Omics Analysis to Investigate ZH8015's Responses Against Brown Planthopper Infestation

Accurate genomic information is essential for advancing genetic breeding research in specific rice varieties.This study presented a gapless genome assembly of the indica rice cultivar Zhonghui 8015(ZH8015)using Pac Bio HiFi,Hi-C,and ONT(Oxford Nanopore Technologies)ultra-long sequencing technologies,annotating 43037 gene structures.Subsequently,utilizing this genome along with transcriptomic and metabolomic techniques,we explored ZH8015's response to brown planthopper(BPH)infestation.Continuous transcriptomic sampling indicated significant changes in gene expression levels around 48 h after BPH feeding.Enrichment analysis revealed particularly significant alterations in genes related to reactive oxygen species scavenging and cell wall formation.Metabolomic results demonstrated marked increases in levels of several monosaccharides,which are components of the cell wall and dramatic changes in flavonoid contents.Omics association analysis identified differentially expressed genes associated with key metabolites,shedding light on ZH8015's response to BPH infestation.In summary,this study constructed a reliable genome sequence resource for ZH8015,and the preliminary multi-omics results will guide future insect-resistant breeding research.

Developmental Genetic Analysis of Brown Rice Weight Under Different Environmental Conditions in indica Rice

Analysis of genetic main effects and genotype x environment (GE) interaction effects for brown rice weight (BRW) at four different filling stages in indica lice (Oryza sativa L.) was conducted for two-year experimental data by using developmental genetic models and corresponding statistical approaches for quantitative traits of seeds in cereal crops. It was indicated that the genetic main effects and their GE interaction effects of triploid endosperm, cytoplasmic and diploid maternal plant genes were important for BRW at different filling stages of rice, especially for endosperm or maternal additive main effects and their additive interaction effects. Because of the higher additive effects and additive interaction effects for BRW at different filling stages, the better improving effects for this trait could be expected by selection in rice breeding. The results of conditional genetic variance components showed that the new expression of quantitative genes in endosperm and maternal plant for BRW was mostly found at all different filling stages of rice. The gene expression, however, was most active at the early filling stages especially for the first (1-7 d) and the second filling stages (8-14 d after flowering). The phenomena that some genes were spasmodically expressible among filling stages of rice were detected for some genetic effects especially for net cytoplasmic main effects or its interaction effects and net dominance main effects. Predicted genetic effects at different filling stages of rice showed that some parents such as V20 and Zuo 5 were better than others for improving the BRW.

Analysis of Resistance-related Proteins in Rice Against Brown Planthopper by Two-dimensional Electrophoresis

A recombinant inbred population (RI) was constructed from a cross between B5, an introgression. line from the wild rice Oryza officinalis Wall. ex Watt, and susceptible cultivar Minghui 63 ( O. sativa L.). The brown planthopper ( BPH) resistances of RI lines were evaluated. Based on bulked segregant analysis (BSA), two protein bulks were made by extracting proteins from equally mixed seedlings of extremely resistant and susceptible plants selected from the RI population, respectively. Two-dimensional electrophoresis was used to detect the changes of polypeptide pattern. Results showed that a protein P40 ( pI 6.3, Mw 40 kD) was significantly reduced or vanished after BPH infestation for 48 h in the susceptible bulk, while it remained uninfluenced in the resistant bulk. In connection with the physiological changes of the resistant and susceptible lines subjected to BPH sucking, we suppose that the protein P40 is related to the interaction responses of lice plants to BPH infestation.

Effects of Storage Temperature on Pasting Properties of Brown Rice

[Objective] The aim of this study was to explore the effects of storage temperature on pasting properties of brown rice.[Method] Rapid viscosity analyzer was used to determine the viscosity of brown rice under different storage temperatures（30,25,20 and 15 ℃）.[Result] The peak viscosity value increased with storage time prolonging.The higher the storage temperature was,the more rapidly the increasing of peak viscosity value;different storage temperatures and storage time showed significant effects on peak viscosity value of brown rice;the difference on peak viscosity value of brown rice under different storage temperatures was significant.The hot viscosity value of brown rice stored under 30 and 25 ℃ gradually increased with the storage time prolonging;and storage time and storage temperature showed significant effects on hot viscosity value;there was extremely significant difference on hot viscosity value of brown rice under 30,25 and 20,15 ℃.The breakdown value showed a first increasing and then decreasing trend with the storage time prolonging;storage temperature and storage time had extremely significant effect on the breakdown value of brown rice;the breakdown value of brown rice among each storage temperature was extremely significant.The final viscosity of brown rice increased with storage time prolonging.The higher the temperature was,the higher the increasing rate of final viscosity;storage time and temperature showed extremely significant effect on the final viscosity of brown rice;the difference on final viscosity among different storage temperature treatments was extremely significant.The changes on setback value of brown rice under different storage temperature treatments were different with the storage time prolonging;and the storage temperature showed extremely significant effect on the setback value of brown rice.[Conclusion] The study had provided theoretical basis for the storage of brown rice.

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

Density Restriction Effects of Reproductive Rate of Rice Brown Planthopper Population in Super Rice Yongyou 6

[ Objective ] The paper was to study the density restriction effects of reproductive rate of rice brown planthopper （BPH） population in super indiea and japonica hybrid rice Yongyou 6. [Method] Different amounts of rice brown planthoppor were released to super indiea and japonica hybrid flee Yongyou 6 during fillering, booting and gain filling stage, respectively. The reproductive rate and variation dynamics of rice brown planthoppor in super rice was also investigated and analyzed. [Result] Tillering stage： the population quantity of rice brown planthoppor during booting and gain filling stage gradually increased with the increasing basal amount of its population during tillering stage; when the population density of rice brown planthopper during grain filling stage reached a certain limit （ 〉 250 head/cluster）, the population density during milking stage showed decrease trend with the increasing of such density; the development of rice brown planthoppor population in super rice Yongyou 6 showed the characteristics of fast in middle and slow in two ends. Booting stage： the development of rice brown planthopper pop- ulation from booting to gain filling stage increased with the increasing basal amount of density, and their correlation during milking stage was not significant; flee brown planthopper population still had high reproduction capacity in the middle and late stage of super rice, which had latent risk on rice yield. Gain filling stage ： when population density of rice brown planthopper 〈 70 head/cluster, the development of it population increased with the increase of basal amount of density, which had great threat to yield. [ Conclusion] The result provided theoretical basis for establishment and improvement of monitoring early warning system, cost-ef- fective control of occurrence and damage of rice brown planthopper, and improvement of the sustained control ability against rice brown planthopper.

Identification of an anticancer compound against HT-29 cells from Phellinus linteus grown on germinated brown rice

Objective:To isolate and identify the anticancer compound against proliferation of human colon cancer cells from ethyl acetate(EtOAc)extract ol Phellinus linteus grown on germinated brown rice(PB).Methods:EtOAc extract of PB was partitioned with n-hexane,EtOAc,and water-saturated n-butanol.Anticancer compound of n-hexane layer was isolated and identified by HPLC and NMR,respectively.Cytotoxicity against HT-29 cells was tested by SRB assay.Results:The n-hexane layer obtained after solvent fractionation of PB EtOAc extracts showed a potent anticancer activity against the HT-29 cell line.Atractylenolide I,a eudesmane-type sesquiterpene lactone,a major anticancer substance of PB,was isolated from the n-hexane layer by silica gel column chromatography and preparative-HPLC.This structure was elucidated by one-and two-dimensional NMR spectroscopic data.Atractylenolide I has not been reported in mushrooms or rice as of yet.The isolated compound dose-dependently inhibited the growth of HT-29 human colon cancer cells.Conclusions:Atractylenolide I might contribute to the anticancer effect of PB.

Characterizing and estimating rice brown spot disease severity using stepwise regression,principal component regression and partial least-square regression

Detecting plant health conditions plays a key role in farm pest management and crop protection. In this study, measurement of hyperspectral leaf reflectance in rice crop (Oryzasativa L.) was conducted on groups of healthy and infected leaves by the fungus Bipolaris oryzae (Helminthosporium oryzae Breda. de Hann) through the wavelength range from 350 to 2 500 nm. The percentage of leaf surface lesions was estimated and defined as the disease severity. Statistical methods like multiple stepwise regression, principal component analysis and partial least-square regression were utilized to calculate and estimate the disease severity of rice brown spot at the leaf level. Our results revealed that multiple stepwise linear regressions could efficiently estimate disease severity with three wavebands in seven steps. The root mean square errors (RMSEs) for training (n=210) and testing (n=53) dataset were 6.5% and 5.8%, respectively. Principal component analysis showed that the first principal component could explain approximately 80% of the variance of the original hyperspectral reflectance. The regression model with the first two principal components predicted a disease severity with RMSEs of 16.3% and 13.9% for the training and testing dataset, respec-tively. Partial least-square regression with seven extracted factors could most effectively predict disease severity compared with other statistical methods with RMSEs of 4.1% and 2.0% for the training and testing dataset, respectively. Our research demon-strates that it is feasible to estimate the disease severity of rice brown spot using hyperspectral reflectance data at the leaf level.

Induced defense responses in rice plants against small brown planthopper infestation

The small brown planthopper(SBPH), Laodelphax striatellus Fallén(Homoptera: Delphacidae), is a serious pest of rice(Oryza sativa L.) in China. To understand the mechanisms of rice resistance to SBPH, defense response genes and related defense enzymes were examined in resistant and susceptible rice varieties in response to SBPH infestation. The salicylic acid(SA) synthesis-related genes phenylalanine ammonia-lyase(PAL), NPR1, EDS1 and PAD4 were induced rapidly and to a much higher level in the resistant variety Kasalath than in the susceptible cultivar Wuyujing 3 in response to SBPH infestation. The expression level of PAL in the Kasalath rice at 12 h post-infestation(hpi) increased 7.52-fold compared with the un-infested control, and the expression level in Kasalath was 49.63, 87.18, 57.36 and 75.06 times greater than that in Wuyujing 3 at 24, 36, 48 and 72 hpi, respectively. However, the transcriptional levels of the jasmonic acid(JA) synthesis-related genes LOX and AOS2 in resistant Kasalath were significantly lower than in susceptible Wuyujing 3 at 24, 36, 48 and 72 hpi. The activities of the defense enzymes PAL, peroxidase(POD), and polyphenol oxidase(PPO) increased remarkably in Kasalath in response to SBPH infestation, and were closely correlated with the PAL gene transcript level. Our results indicated that the SA signaling pathway was activated in the resistant Kasalath rice variety in response to SBPH infestation and that the gene PAL played a considerable role in the resistance to SBPH.

Expression Analysis of Genes Related to Rice Resistance Against Brown Planthopper, Nilaparvata lugens

Brown planthopper(BPH) is an insect species that feeds on the vascular system of rice plants. To examine the defence mechanism of rice plants against BPH, the pathogenesis-related genes(PR1a, PR2, PR3, PR4, PR6, PR9, PR10a, PR13, PR15 and PRpha), signaling molecule synthesis genes(AOS, AXR, ACO and LOX), antioxidant-related genes(CAT, TRX, GST and SOD) and lignin biosynthesis-related genes(CHS, CHI and C4H) were investigated in a resistant rice variety. AOS, PR6,PR9 and PR15 genes showed significantly increased relative expression levels at 24.38-, 19.17-, 14.71-, and 12.74-fold compared to the control. Moderate increased relative expression levels of lignin biosynthesis-related gene(C4H), pathogenesis-related genes(PR4, PR10a and PRpha), and antioxidant-related gene(GST) were found, while CHI, LOX, SOD, TRX1 and AXR showed decreased relative expression levels. It was thus clearly shown that wound-induced response genes were activated in rice plants after BPH attacks through AOS activation. Jasmonic acid signaling molecule may activate PR6, PR15, GST and CAT subsequently increasing their expression for H_2O_2 detoxification. PR6 were expressed at the highest relative level among the PR genes. These genes therefore have also a considerable synergistic role with the other genes against BPH by interfered their digestion tract system.

Pyramiding blast,bacterial blight and brown planthopper resistance genes in rice restorer lines

Rice blast, bacterial blight （BB） and brown planthopper （BPH） are the three main pests of rice. This study investigated pyr-amiding genes resistant to blast, BB and BPH to develop restorer lines. Ten new lines with blast, BB and/or BPH resistance genes were developed using marker-assisted selection （MAS） technique and agronomic trait selection （ATS） method. Only HR13 with resistance genes to blast, BB and BPH was obtained. In addition to blast and BB resistance, four lines （HR39, HR41, HR42, HR43） demonstrated moderate resistance to BPH, but MAS for BPH resistance genes were not conducted in developing these four lines. These data suggested that there were unknown elite BPH resistance genes in the Zhongzu 14 donor parent. A more effective defense was demonstrated in the lines withPi1 andPi2 genes although the weather in 2012 was favorable to disease incidence. Blast resistance of the lines with a single resistance gene,Pita, was easily inlfuenced by the weather. Overal, the information obtained through pyramiding multiple resistance genes on developing the restorer lines is helpful for rice resistance breeding.

Donors for Resistance to Brown Planthopper Nilaparvata lugens(St?l) from Wild Rice Species

Out of 1 989 wild accessions sown in seed boxes for screening, only 1 003 wild accessions with good germination were screened against brown planthopper(BPH), Nilaparvata lugens(St?l) under greenhouse conditions. The collection comprised of accessions from 11 wild species and African cultivated rice. The germplasm was screened for BPH following standard seed box screening technique in the greenhouse. As many as 159 accessions were identified as resistant during the year 2012 based on one year screening. A selected set of BPH resistant accessions were screened again during 2013. Based on the two years screening, seven accessions of O. nivara(AA), one accession of O. officinalis(CC), seven accessions of O. australiensis(EE), five accessions of O. punctata(BB and BBCC) and nine accessions of O. latifolia(CCDD) were confirmed to be resistant to BPH. So far no BPH resistance genes have been identified and designated from O. nivara and O. punctata, hence these may act as new sources of resistance.

LncRNAs are potentially involved in the immune interaction between small brown planthopper and rice stripe virus

Small brown planthopper(SBPH, Laodelphax striatellus Fallén) is an important vector of major crop pathogen rice stripe virus(RSV). Controlling SBPH population is an efficient approach to control RSV. Long non-coding RNAs(lnc RNA) have been reported to block virus replication in hosts. However, the function of lnc RNAs in RSV infection and replication is still unknown. Here, we aimed to study regulatory mechanisms of lnc RNA in an immune system during RSV infection. First, lnc RNA genes were predicted from SBPH transcriptomes using a bioinformatics pipeline based on characteristics of lnc RNA. We identified 4 786 lnc RNA genes corresponding to 5 790 transcripts in SBPH from an RNA-Seq dataset of 15 transcriptomes. Differential expression analysis indicated that 3, 11, and 25 lnc RNA genes were highly expressed in gut, salivary gland, and ovary, respectively, of viruliferous SBPH(Student’s t-test, P<0.05). We randomly selected eight lnc RNAs for expression validation using quantitative real-time PCR, confirming the differential expression of these lnc RNAs between viruliferous and non-viruliferous SBPH. In summary, we present evidence that the expression of lnc RNA genes was induced by RSV infection, suggesting that RSV might be involved in the antivirus immune system in SBPH and participate in regulating the RSV replication mechanism. These data provide helpful information for future investigations of the interaction between lncRNA and RSV.

Study on the Optimum Moisture Amount Added to Brown Rice once during the Wet Conditioning

In the experiment, the brown rice whose moisture content was 12.5% was used as raw material. The brown rice was grouped, then moisturized differently and milled. While milling, the energy consumption, the rate of broken rice and the crack rate were tested. It is confirmed that the stress crack owing to the moisture added to the brown rice can be avoided when the moisture amount added once is limited to no more than 1.5%. It is also proved that the energy consumption can be reduced, the yielding rate of rice can be increased and that the quality of rice can be improved.

Highly Sensitive and Specific Monoclonal Antibody-Based Serological Methods for Rice Ragged Stunt Virus Detection in Rice Plants and Rice Brown Planthopper Vectors

Rice ragged stunt virus（RRSV） is a serious rice disease in Asia, causing serious yield losses on rice. The capsid protein（CP） gene of the major outer capsid protein of RRSV was expressed in Escherichia coli BL21（DE3） using the pMAL-C2 X expression vector. The recombinant protein was used as the immunogen to immunize BALB/c mice. A hybridoma cell line 8A12 secreting monoclonal antibody（MAb） against RRSV was obtained by fusing mouse myeloma cells（Sp 2/0） with spleen cells from the immunized BALB/c mice. Western blot analysis showed that the MAb 8A12 can specifically react with RRSV CP. Using the MAb, an antigen-coated-plate enzyme-linked immunosorbent assay（ACP-ELISA）, a dot enzyme-linked immunosorbent assay（dot-ELISA）, and immunocapture-RT-PCR（IC-RT-PCR） assay were developed to detect RRSV. The established ACP-ELISA, dot-blot ELISA and IC-RT-PCR methods could detect RRSV in infected rice tissue crude extracts with dilutions of 1：40 960, 1：1 280 and 1：655 360（w/v, g mL-1）, respectively. The ACP-ELISA and dot-blot ELISA methods could detect RRSV in infected insect vector crude extracts with dilutions of 1：12 800 and 1：1 600（an individual planthopper μL-1）, respectively. The field survey revealed that Rice ragged stunt disease occurs on rice in Hainan, Yunnan, Guangxi, Sichuan, Guizhou, Fujian, Hunan, Jiangxi and Zhejiang in China.

Evaluation of Rice Germplasm for Resistance to the Small Brown Planthopper (Laodelphax striatellus) and Analysis of Resistance Mechanism

One hundred and thirty-eight rice accessions were screened for resistance to the small brown planthopper （SBPH） （Laodelphax striatellus Fallen） by the modified seedbox screening test. Twenty-five rice accessions with different levels of resistance to SBPH were detected, accounting for 18.1% of the total accessions, which included 2 highly resistant, 9 resistant and 14 moderately resistant varieties. Compared with indica rice, japonica rice was more susceptible to SBPH. Antixenosis test, antibiosis test and correlation analysis were performed to elucidate the resistance mechanism. The resistant check Rathu Heenati （RHT）, highly resistant varieties Mudgo and Kasalath, and resistant variety IR36 expressed strong antixenosis and antibiosis against SBPH, indicating the close relationship between resistance level and these two resistance mechanisms in the four rice varieties. Antibiosis was the dominant resistance pattern in the resistant varieties Daorenqiao and Yangmaogu due to their high antibiosis but low antixenosis. Dular, ASD7 and Milyang 23 had relatively strong antixenosis and antibiosis, indicating the two resistance mechanisms were significant in these three varieties. The resistant DV85 expressed relatively high level of antixenosis but low antibiosis, whereas Zhaiyeqing 8 and Guiyigu conferred only moderate antibiosis and antixenosis to SBPH, suggesting tolerance in these three varieties. Antibiosis and antixenosis governed the resistance to SBPH in the moderately resistant accession 9311. Antixenosis was the main resistance type in V20A. Tolerance was considered to be an important resistance mechanism in Minghui 63 and Yangjing 9538 due to their poor antibiosis and antixenosis resistance. The above accessions with strong antibiosis or antixenosis were the ideal materials for the resistance breeding.

Transmission of Rice Black-Streaked Dwarf Virus from Frozen Infected Leaves to Healthy Rice Plants by Small Brown Planthopper (Laodelphax striatellus)

In order to preserve virus for identifying the resistance of rice varieties against rice black-streaked dwarf disease, a simple and reliable method was developed, through which virus-free small brown planthopper （SBPH） acquired rice black-streaked dwarf virus （RBSDV） from frozen infected leaves and the virus was transmitted to healthy rice plants. The experimental results showed that SBPH could obtain RBSDV from frozen infected rice leaves and the virus could be transmitted to a susceptible rice variety. For the ability to acquire RBSDV and transmit the virus to healthy plants by SBPH, there was no significant difference between frozen infected leaves and in vitro infected leaves. The novel method could be applied to identification of rice variety resistance to rice black-streaked dwarf disease, facilitating the breeding process for rice black-streaked dwarf disease resistance.

Study on the Relationship between.the Moisture Content of Brown Rice and Milling Characteristics

An experiment about rice milling was made to study on the relationship between the different moisture content of brown rice and the energy consumption, the broken rice rate, the crack rate and the head rice yield. It could be concluded that the head rice yield increased at first falls and then along with the raise of moisture content and it could reach the maximum 70.78% when moisture content was 15.5%; the energy consumption of rice milling decreased along with the increase of moisture contents; the broken rice rate fell at first and then increased along with the raise of moisture contents and it could reach the minimum 4.28% when the moisture content was 15.5%.