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[Developmental genetic analysis of brown rice thickness of indica rice (Oryza sativa L.)].

Analysis of developmental genetic effects for brown rice thickness (BRT) trait in indica rice (Oryza sativa L.) at four different filling stages was conducted and a developmental genetic model and corresponding statistical approaches for quantitative traits of triploid endosperm in cereal crops was used. The results indicated that the diploid maternal plant, triploid endosperm and cytoplasmic genetic effects were important for BRT trait at all filling stages of rice and those effects were the major effect at initial medium, filling period and mature period respectively. The additive and dominance effects were the major effect alternatively at four filling stages of rice. Significant endosperm and maternal dominance effects for BRT suggest that the utilization of heterosis for BRT is viable. The results of conditional genetic variance components shown that the expression of new quantitative genes in endosperm, cytoplasm and maternal plant for BRT was found at most filling stages of rice. The gene expression was most active at the early filling stages, especially at the second stage (8-14 days after flowering). Near to mature period (22-28 days after flowering), however, the expression of genes decreased sharply and even closed. The phenomena that the genes expressed spasmodically, i.e. the net genetic effects equaled to zero in some filling stages were detected for some genetic effects. Dominance and cytoplasmic correlation coefficients were significant at 0.05 or 0.01 probability level in some filling stages meanwhile there were the most strong relationship between mature period and other periods for BRT.

Genetic Variation↗

ICON rice seed treatment toxicity to crayfish (Procambarus clarkii) in experimental rice paddies.

Outdoor pools (2.3 x 2.3 m) were used to simulate typical rice agricultural practices in Louisiana, USA, to evaluate the toxicity of ICON (active ingredient [a.i.] fipronil) and its degradates to crayfish (Procambarus clarkii). Six paddies were planted with seed treated with ICON 6.2 FS at an exaggerated application rate of 0.05 kg a.i./ha (recommended rate, 0.042 kg a.i./ha), simulating three rice-planting scenarios. Two reference paddies were planted with untreated seed. Crayfish were exposed to tail water within 24 to 48 h after seeding, simulating standard Louisiana agricultural and water management practices. At 50 d after planting, a separate group of crayfish was caged in situ for 14 d to evaluate toxicity. An additional 50 crayfish were added to two paddies approximately 100 d after rice planting and held for 29 weeks to evaluate bioaccumulation. Residues of fipronil and its degradates in water and soil were similar to residue concentrations measured from rice fields in Louisiana. Tail water from the treated paddies was not toxic to crayfish. The fipronil 96-h median lethal concentration (LC50) for adult crayfish was 180 microg/L, which would provide at least a sixfold safety factor between the maximum fipronil concentration in tail water and the crayfish LC50. In situ exposures of crayfish also were not toxic. Concentrations of fipronil and its degradates after 29 weeks of exposure were less than 5 microg/kg in crayfish tail muscle tissue. These results demonstrate that label instructions adequately protect crayfish in a rice-crayfish cropping scenario when ICON is applied at maximum application rates as a seed treatment.

Agriculture↗

[Effect of rice-duck mutualism on nutrition ecology of paddy field and rice quality].

The production of nuisanceless rice was conducted by the technique of rice-duck mutualism without applying any pesticide and chemical fertilizer. The results showed that compared to the control, the effect of eliminating weeds was over 99.4%, and the cardinal numbers of plant diseases and insect pests reduced obviously. Nutrients and dissolved oxygen in paddy water and rapidly available nutrients in soil increased, but after maturing stage, compared to basal fertility, rapidly available P and K decreased. The N, P, and K absorption amounts of rice plant, rice yield, setting panicles rate, filled grains and ripened rate increased. The milling, appearance, nutritional, cooking and eating quality of rice improved, especially in reducing chalkiness. The comprehensive benefits of paddy field enhanced obviously.

Animals↗

Genetic analysis of rice varietal diversity for rice blast control.

Two Indica hybrid rice of Shanyou63 (A) and Shanyou22 (B), two glutinous landraces of Huanghenuo (C) and Zinuo (D) and three improved Japonica rice of Hexi41 (E), Chujing12 (F) and 8126 (G) were selected and their genetic resistance relationship was estimated using resistance gene analogue (RGA). The results showed that there were similar genetic relationships between hybrid varieties at the genetic similarity (GS) of 0.86,and among improved Japonica varieties at the GS of 0.84, while highly genetic diversifications between traditional varieties, Indica and Japonica varieties, traditional and modern variety ( GS:0.45). The results also showed that clustering analysis based on RGA data were generally corresponded to known pedigrees and blast field resistances of the varieties. Based on varietal differences in RGA data and agronomic traits, plot experiments of five mixed-planting combinations of A/C, A/D, B/C, B/D and A/B and two combinations of E/C and E/F/G were conducted in Jianshui and Shiping counties ( Indica rice growing region) and Luxi County (warm Japonica region) in Yunnan Province in past two years, respectively. The results demonstrated that rice blast management was more effective in five mixed-planting combinations of varieties with different genetic backgrounds (GS: 0.45-0.77) than in two combinations with similar genetic relationships (GS: 0.84-0.90), compared with their monocultures. It is evident for the highly susceptible landraces in mixed-planting to achieve disease control, with significant decreases both in incidence and severity. The blast control efficiencies of landraces in different mixture combinations reached to 54.47%-92.18%. The control efficiencies of improved varieties varied from 15.12% to 25.54% in mixture combinations with closed genetic relationship. In addition,the total yield of 5 varietal combinations with distant genetic relationship increased 539.0-904.0 kg/ha in the mixed-planting plots, at increase rates of 5.6%-10.2%. Mixed rice varieties with similar genetic background did not achieve significant yield increase. Otherwise, the yield of E/F/G decreased 2.7%-4.0% compared with pure stand. The results can provide scientific basis of varietal combinations in diversification experiments for blast control.

Genetic Variation↗

Comparison of ileal digested production of parental rice and rice genetically modified with cowpeas trypsin inhibitor.

OBJECTIVE: To compare the ileal digestibility of protein and amino acids in parental rice and rice genetically modified with sck gene. METHODS: Six experimental swines were surgically fixed with a simple T-cannula at the terminal ileum and fed with parental rice and rice genetically modified with sck gene alternately. The ileum digesta were collected and analyzed for determination of apparent and true digestibility of protein and amino acids. RESULTS: The apparent and true digestibility of protein was similar in these two types of rice. Except for the apparent digestibility of lysine, there was no difference in the apparent and true digestibility of the other 17 amino acids. CONCLUSION: The digestibility of protein and amino acids is not changed by the insertion of foreign gene, so it can meet the request of "substantial equivalence" in digestibility of protein and amino acids.

Amino Acids↗

Overexpression of rice OSH genes induces ectopic shoots on leaf sheaths of transgenic rice plants.

Five rice homeobox (OSH) genes were overexpressed under the control of the cauliflower mosaic virus 35S promoter or the rice actin gene promoter in transgenic rice plants. Almost all of the transgenic plants showed abnormal phenotypes, which could be classified into three types according to their severity. Plants with the most severe phenotype formed only green organs, with many shoot apices on their adaxial sides. Plants with an intermediate phenotype formed bladeless leaves with normally developed leaf sheaths. Plants with a mild phenotype formed normal leaf sheaths and blades, but lacked ligules and showed diffusion of the blade-sheath boundary. The leaf structure of this phenotype was similar to that of dominant maize mutants, such as Kn1, Rs1, Lg3, and Lg4. Based on these phenotypes, we suggest that ectopic expression of the rice OSH genes interferes with the development of leaf blades and maintains leaves in less differentiated states. These results are discussed in relation to the leaf maturation schedule hypothesis (M. Freeling et al., 1992, BioEssays 14, 227-236).

Actins↗

Physical mapping of a rice lesion mimic gene, Spl1 , to a 70-kb segment of rice chromosome 12.

The rice lesion mimic mutant spotted leaf 1 ( spl1) was first identified in the rice ( Oryza sativa) cultivar Asahi in 1965. This mutant displayed spontaneous disease-like lesions in the absence of any pathogen, and was found to confer resistance to multiple isolates of rice blast. We employed a map-based cloning strategy to localize the Spl1 gene. A total of ten cleaved amplified polymorphic sequence (CAPS) markers linked to the Spl1 gene were identified and mapped to an 8.5-cM region on chromosome 12. A high-resolution genetic map was developed using these ten CAPS markers and a segregating population consisting of 3202 individuals. A BAC contig containing four BAC clones was constructed, and Spl1 was localized to a 423-kb region. Seven spl1 mutants were obtained from the IR64 deletion mutant collection, and molecular analysis using these mutants delimited the Spl1 gene to a 70-kb interval, covered by two BAC clones. These results provide the basis for cloning this gene, which is involved in cell death and disease resistance in rice.

Genetic Markers↗

The rice OsGAE1 is a novel gibberellin-regulated gene and involved in rice growth.

Gibberellins (GAs) are a class of phytohormones that regulate many aspects of plant growth and development processes including stem elongation, flowering, and seed germination. A novel GA-enhanced gene, designated as OsGAE1, was identified using microarray analysis of GA-regulated genes. OsGAE1 expressed in a dose- and time course-dependent manner with minimum expression at 1 microM GA(3) and maximum expression at 50 microM GA(3) starting from 30 min and peaked at 24 h after GA(3) treatment. OsGAE1 expression was up-regulated by GA(3) at transcript level while no significant effect was observed for other hormones. OsGAE1 was expressed in Escherichia coli with N-terminal His(6) tag and the recombinant protein migrated at 38 kDa, slightly larger than the predicted 29 kDa, during SDS-PAGE. Anti-OsGAE1 antibodies immunoreacted with a protein of 40 kDa in rice leaf sheath. OsGAE1 expressed mainly in growing leaf sheath and callus compared to leaf and root. In situ hybridization and OsGAE1 promoter analysis revealed that OsGAE1 expressed in shoot apex meristem and young primary leaves. Northern blot, Western blot, and GUS activities revealed that OsGAE1 is up-regulated by GA(3). Transgenic rice expressing OsGAE1 in antisense orientation exhibited severely affected vegetative and reproductive growth. The transgenic plants were 55-70% short compared to control. These results suggest that OsGAE1 is differentially expressed in rice leaf sheath in relation to GA(3) and it encodes a functional protein which is involved in GA-regulated growth and development of rice.

Amino Acid Sequence↗

Effects of lead contamination on soil microbial activity and rice physiological indices in soil-Pb-rice (Oryza sativa L.) system.

The effect of lead (Pb) treatment on the soil microbial activities (soil microbial biomass and soil basal respiration) and rice physiological indices were studied by greenhouse pot experiment. Pb was applied as lead acetate at six different levels in two different paddy soils, namely 0 (control), 100, 300, 500, 700, 900 mg kg-1 soil. The results showed that the application of Pb at lower level (<300 mg kg-1) as lead acetate resulted in a slight increase in soil microbial activities compared with the control, and had an inhibitory influence at high concentration (>500 mg Pb kg-1 soil), which might be the critical concentration of Pb causing a significant decline in the soil microbial activities. However, the degree of influence on soil microbial activities by Pb was related to the clay and organic matter contents of the soils. On the other hand, when the level of Pb treatments increased to 500 mg kg-1, there was ecological risk for both soil microbial activities and plants. The results also revealed that there was a consistent trend that the chlorophyll contents increased initially, and then decreased gradually with increase in Pb concentration. Pb was effective in inducing proline accumulation and its toxicity causes oxidative stress in rice plants. In a word, soil microbial activities and rice physiological indices, therefore, may be sensitive indicators reflecting environmental stress in soil-Pb-rice system.

Biomass↗

A validated stability-indicating HPLC with photodiode array detector (PDA) method for the stress tests of Monascus purpureus-fermented rice, red yeast rice.

A stability-indicating reversed phase high-performance liquid chromatography (RP-HPLC) with photodiode array (PDA) detection method was developed and validated for the assay of monacolin series compounds including monacolin K, L, J and their hydroxyl acid forms as well as dehydroxymonacolin K simultaneously in Monascus purpureus-fermented rice, red yeast rice. Well-resolved peaks of seven main compounds of monacolin family were profiled on a C(18) reverse-phase column using a linear gradient of 0.1% trifluoroacetic acid and acetonitrile as the mobile phase, and the detection wavelength was set at 237nm. The method was validated with respect to specificity, chromatographic parameters, linearity, precision, accuracy, limits of detection and quantitation. The stability stress testing for fermented red yeast rice powder was carried out to show the effects of high temperature (80 degrees C), high humidity at room temperature (92.5% RH, 25 degrees C), high humidity at high temperature (75% RH, 60 degrees C) and light (sunlight) in solid state. The results exhibited that monacolins decreased significantly under the conditions of high humidity at high temperature (75% RH, 60 degrees C) and sunlight. Monacolin K and its hydroxyl acid form would be dehydrolyzed and turned to dehydromonacolin K at high temperature (80 degrees C) while the monacolin K, J and L would be transformed into their corresponding hydroxyl acid forms under the condition of high humidity (92.5% RH, 25 degrees C). The indication is that monacolins in red yeast rice powder are light-sensitive and thermal-sensitive. Therefore, it has been suggested that the preparations containing monacolins be stored in the place of cool and lightproof. The proposed degradation pathways were discussed as well. The multi-components assay for stability of botanical products could provide much more information than the normal marker-orientation method.

Chromatography, High Pressure Liquid↗

Purification of the rice embryo lectin and its binding to nitrogen-fixing bacteria from the rhizosphere of rice.

A lectin was purified from rice embryos by aqueous acid extraction of crude embryo powder, followed by ammonium sulfate precipitation, affinity chromatography on agarose p-aminophenyl-beta-D-N-acetylglucosamine and gel-filtration on AcA 54. Its homogeneity was checked by polyacrylamide gel electrophoresis, gel-filtration and immunological methods. The hemagglutinating activity of the purified rice lectin was 0.02 micrograms/ml. This lectin labelled with [14C] acetic anhydride was shown to interact in vitro with different bacteria isolated from the rhizosphere of rice. The most efficient binding was obtained with Beijerinckia V.. The affinity constant Ka was (1.04 +/- 0.30) X 10(7) M-1 and each bacterium contained 1660 +/- 150 lectin receptor sites. In contrast, no interaction between bacteria isolated from the rhizosphere of maize or E. coli K 12 and rice lectin was evidenced.

Agglutination Tests↗

Pathway of CH4 formation in anoxic rice field soil and rice roots determined by 13C-stable isotope fractionation.

In anoxic rice fields methane is produced by either reduction of CO2 or cleavage of acetate. We measured the delta 13C-values of CH4 and CO2, acetate and organic carbon during time course experiments with anoxic methanogenic soil and root samples and used these values to calculate the fractions of CH4 (and acetate) produced from CO2 reduction. Comparison with radiotracer and/or inhibitor studies constrained the kinetic fractionation factors used for calculation. The fractionation factors for the conversion of CO2 to CH4 and of acetate to CH4 were on the order of alpha = 1.07 (epsilon = -70%) and epsilon > or = - 20%, respectively. The pathway of CH4 production changed with time of anoxic incubation. Anoxic slurries of rice field soil first produced CH4 predominantly (>50%) from CO2, then predominantly (>80%) from acetate and finally (after about one month) according to the theoretically expected ratio (33% CO2 and 67% acetate). Anoxic rice roots, on the other hand, initially produced CH4 exclusively from CO2, followed by contribution of acetate of about 40-60%. Rice roots also produced acetate that partially originated (< or = 1 30%) from reduction of CO2 as determined by calculation of isotopic fractionation using fractionation factors from the literature. The results demonstrate that there is quite some variability in pathways of CH4 production, and also indicate that isotopic fractionation factors may be different in different habitats and change with time.

Carbon Dioxide↗

Enhanced resistance to sheath blight by constitutive expression of infection-related rice chitinase in transgenic elite indica rice cultivars.

Genetic transformation has been attempted for management of rice sheath blight disease, caused by Rhizoctonia solani. We introduced a PR-3 rice chitinase gene (RC7), isolated from R. solani-infected rice plants, into indica rice cultivars IR72, IR64, IR68899B, MH63, and Chinsurah Boro II by the biolistic and PEG-mediated transformation system. Inheritance was studied up to the T(2) generation by Southern blot analysis. Western blot analysis of transgenic plants with polyclonal antibody revealed the presence of chitinase protein with a molecular weight of 35 kDa that reacts with chitinase antibody. The transformants synthesized different levels of chitinase proteins constitutively and progeny from the plants containing the chitinase gene showed different levels of enhanced resistance when challenged with the sheath blight pathogen R. solani.

Journal Article↗

Formate dehydrogenase in rice plant: growth stimulation effect of formate in rice plant.

NAD-dependent formate dehydrogenase (FDH, EC 1.2.1.2.) plays a key role in the final step of the catabolism of methanol in methylotrophs. In certain plant species, this enzyme is a mitochondrial and NAD-dependent enzyme. In this study, the complete cDNA sequence of the rice (Oryza sativa cv Hinohikari) FDH gene, which contains an open reading frame (ORF) of 1128 bp, was determined. The sequence corresponds to a protein of 376 amino acids and shows strong homology to the NAD-dependent FDHs from other plants. Our experimental results showed that the expression of the rice cDNA in Escherichia coli induced FDH activity, indicating that the cDNA encodes a functional peptide of FDH. The application of formate to the roots of rice seedlings leads to growth increment both in terms of the length of the aerial part and the fresh weight. The FDH activity and the accumulation of FDH transcripts were strongly enhanced in the root portions of the formate-fed plants. This enhancement indicates that the transcriptional regulation which responds to formate functions in the rice plant, and that formate can act as an effective inducer for FDH activity.

Journal Article↗

Extraction of fumonisins B1 and B2 from white rice flour and their stability in white rice flour, cornstarch, cornmeal, and glucose.

To extract fumonisin B1 (FB1) and fumonisin B2 (FB2) from Thai white rice flour, different solvent mixtures, temperatures, pH values, and addition of enzymes or ethylenediaminetetraacetic acid disodium salt (Na2EDTA) were examined. Three extractions with 0.1 M Na2EDTA achieved the highest recoveries. Initial recoveries of fumonisins added to white rice flour, cornstarch, cornmeal, and glucose varied with commodity. Fumonisins disappeared in Thai white rice flour after 12 h, but 55% remained in another white rice flour. With cornstarch 20-30% fumonisins remained after 24 h; only 43% of 14C-labeled FB1 materials extracted from cornstarch was eluted with methanol from an immunoaffinity column. Fumonisins were stable in cornmeal for 24 h but only approximately 50% remained after 30 days. With glucose, 25% of FB1 and FB2 remained 24 h after addition; N-(1-deoxy-D-fructos-1-yl)FB(1) andN-(carboxymethyl)FB(1) were detected in lower amounts than residual FB(1) after 3 months.

Carboxylic Acids↗

Fibrillar microfilaments associated with a high-amylose rice, goami 2, a mutant of ilpumbyeo, a high-quality japonica rice.

The ultrastructure of cooked and malt-treated cooked rice of Ilpumbyeo (IP) and its mutant Goami 2 (G2), which have extreme contrasts in physicochemical properties, cooking quality, and ultrastructural characteristics in raw grains (1, 2), was compared. In cooked rice of IP, starch granules in endosperm cells were evenly coalesced, appearing as homogeneously smooth sheetlike matrix and/or globules, whereas those in G2 were a heterogeneously coarse matrix in which a novel structural feature, the microfilaments, was embedded. In malt-treated cooked rice of IP, most starch was hydrolyzed by the malt enzymes, appearing as empty vacuoles surrounded by the cell wall, whereas that in G2 was highly resistant to malt treatment, remaining as distinct structural features, the malt-resistant compound starch granules. The property of G2's compound starch granules, which are tolerant of mechanical and chemical treatments thereby retaining their structural integrity (2) and of cooking and malt treatment thereby retaining their physical hardness, appears to play a major role in determining the quality of cooked rice of G2.

Actin Cytoskeleton↗

Resistance to rice yellow mottle virus (RYMV) in cultivated African rice varieties containing RYMV transgenes.

The disease caused by rice yellow mottle virus (RYMV) is a serious problem for African rice growers in large-scale irrigated programs. As there are very few suitable natural sources of RYMV resistance, we have investigated a transgenic approach using widely grown, RYMV-susceptible cultivars of rice and a transgene encoding the RNA-dependent RNA polymerase of RYMV. Transformed lines were resistant to RYMV strains from different African locations. In the most extreme examples there was complete suppression of virus multiplication. Resistance was stable over at least three generations. Subject to satisfactory field testing, these transgenic lines may be suitable for introduction into RYMV-affected rice-growing areas. In the most resistant line, transcription analysis indicated that the resistance derives from an RNA-based mechanism associated with posttranscriptional gene silencing.

Genome, Viral↗

The regulatory regions of the rice tungro bacilliform virus promoter and interacting nuclear factors in rice (Oryza sativa L.).

The Rice Tungro Bacilliform Virus (RTBV) promoter confers phloem-specific gene expression in transgenic rice plants. A series of promoter deletion mutants were fused with the Escherichia coli beta-glucuronidase A (uidA) reporter gene and introduced into transgenic rice plants. The RTBV promoter confers substantially stronger expression in shoots than in roots. A fragment of the promoter comprising nucleotides -164 to +45 relative to the transcriptional start site contains sufficient information for phloem-specific gene expression. Within this region, nucleotides -164 to -43 were essential for promoter function since deletion of this fragment dramatically reduced promoter activity. Gel-retardation assays identified two groups of rice nuclear factors (RNFG1 and RNFG2) that bind to the -164 to +45 promoter fragment. Competition and DNasel footprinting experiments indicated that RNFG1 bound to nucleotides -3 to +8 (Box I) while RNFG2 bound to nucleotides -53 to -39 (Box II). Interactions between the two groups of factors were observed. In addition, we found differences in the binding of nuclear factors from shoots versus from roots, in agreement with the different activities of the promoter in these two organs. It is proposed that binding of RNFG1 and RNFG2 between nucleotides -164 to +45 is essential for the tissue-specific expression of this promoter.

Base Sequence↗