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Sequence analysis of the long arm of rice chromosome 11 for rice-wheat synteny.

The DNA sequence of 106 BAC/PAC clones in the minimum tiling path (MTP) of the long arm of rice chromosome 11, between map positions 57.3 and 116.2 cM, has been assembled to phase 2 or PLN level. This region has been sequenced to 10x redundancy by the Indian Initiative for Rice Genome Sequencing (IIRGS) and is now publicly available in GenBank. The region, excluding overlaps, has been predicted to contain 2,932 genes using different software. A gene-by-gene BLASTN search of the NCBI wheat EST database of over 420,000 cDNA sequences revealed that 1,143 of the predicted rice genes (38.9%) have significant homology to wheat ESTs (bit score >/= 100). Further BLASTN search of these 1,143 rice genes with the GrainGenes database of sequence contigs containing bin-mapped wheat ESTs allowed 113 of the genes to be placed in bins located on wheat chromosomes of different homoeologous groups. The largest number of genes, about one-third, mapped to the homoeologous group 4 chromosomes of wheat, suggesting a common evolutionary origin. The remaining genes were located on wheat chromosomes of different groups with significantly higher numbers for groups 3 and 5. Location of bin-mapped wheat contigs to chromosomes of all the seven homoeologous groups can be ascribed to movement of genes (transpositions) or chromosome segments (translocations) within rice or the hexaploid wheat genomes. Alternatively, it could be due to ancient duplications in the common ancestral genome of wheat and rice followed by selective elimination of genes in the wheat and rice genomes. While there exists definite conservation of gene sequences and the ancestral chromosomal identity between rice and wheat, there is no obvious conservation of the gene order at this level of resolution. Lack of extensive colinearity between rice and wheat genomes suggests that there have been many insertions, deletions, duplications and translocations that make the synteny comparisons much more complicated than earlier thought. However, enhanced resolution of comparative sequence analysis may reveal smaller conserved regions of colinearity, which will facilitate selection of markers for saturation mapping and sequencing of the gene-rich regions of the wheat genome.

Base Sequence↗

Identification and characterization of genes differentially expressed in the resistance reaction in wheat infected with Tilletia tritici, the common bunt pathogen.

The differentially virulent race T1 of common bunt (Tilletia tritici) was used to inoculate the wheat lines Neepawa (compatible) and its sib BW553 (incompatible) that are nearly isogenic for the Bt-10 resistance gene. Inoculated crown tissues were used to construct a suppression subtractive hybridization (SSH) cDNA library. Of the 1920 clones arrayed from the SSH cDNA library, approximately 10 % were differentially regulated. A total of 168 differentially up-regulated and 25 downregulated genes were identified and sequenced; 71 % sequences had significant homology to genes of known function, of which 59 % appeared to have roles in cellular metabolism and development, 24 % in abiotic/biotic stress responses, 3 % involved in transcription and signal transduction responses. Two putative resistance genes and a transcription factor were identified among the upregulated sequences. The expression of several candidate genes including a lipase, two non-specific lipid transfer proteins (ns-LTPs), and several wheat pathogenesis-related (PR)-proteins, was evaluated following 4 to 32 days postinoculation in compatible and incompatible interactions. Results confirmed the higher overall expression of these genes in resistant BW553 compared to susceptible Neepawa, and the differential up-regulation of wheat lipase, chitinase and PR-1 proteins in the expression of the incompatible interaction.

Gene Expression Profiling↗

Physical arrangement of retrotransposon-related repeats in centromeric regions of wheat.

Cereal centromeres commonly contain many repetitive sequences that are derived from Ty3/gypsy retrotransposon. FISH analysis using a large DNA insert library of wheat identified a 67-kb clone (R11H) that showed strong hybridization signals on the centromeres. The R11H clone contains Ty3/gypsy retrotransposon-related sequences; both integrase and CCS1 family sequences were identified. Subsequently, we isolated additional 23 large-insert clones which also contained the integrase and CCS1 sequences. Based on the number of the integrase repeats in the clones determined by DNA gel blot analysis, we concluded that the retrotransposon-like sequences are tandemly repeated in wheat centromeres in ca. 55-kb interval on average. This conclusion is consistent with the results of FISH analysis on the extended DNA fibers.

Base Sequence↗

Assessment of exposure to wheat flour and the shape of its relationship with specific sensitization.

OBJECTIVES: Dust and wheat-allergen exposure were assessed among bakers, flour millers, and bakery-ingredient producers, and the risk for sensitization was studied. METHODS: About 520 inhalable dust and wheat-allergen measurements were made among 270 Dutch workers. Data on sensitization to wheat and common allergens (atopy) were also available. Exposure was estimated according to the sector of industry, job title, and tasks. The shape of the relationship between sensitization and exposure was studied using a two-stage modeling approach: semi-parametric generalized additive modeling and, consequently, a simple description of the relationship using a parametric logistic model. To reduce the effect of exposure measurement errors (attenuation), a combination of the actual measured exposure and variance-weighted estimates of exposure was used. Results The effect of exposure to both inhalable dust and wheat allergens on sensitization was described best by a linear relationship in three industries and a quadratic relationship in one industry. The relation for the whole study population was best described as quadratic, and the probability of sensitization increased with exposure up to -2.7 mg/m3 for inhalable dust and approximately 25.7 microg EQ/m3 for wheat allergens. The risk decreased at higher exposures (P = 0.0121 and P = 0.0731 for dust and wheat, respectively). Atopy and sector of industry modified the sensitization risk significantly in all the analyses. Using a variant-weighted estimator to calculate exposures corrected for the bias and resulted in almost the same point risk estimators. Conclusions Exposure-response relationships for allergens may be nonlinear and differ between industries. A threshold is not indicated on which to base occupational exposure standards; alternatively, other approaches, such as benchmarking, seem warranted.

Allergens↗

Symptom and personality profiles of young adults from a college student population with self-reported illness from foods and chemicals.

Despite much debate over a presumptively somatic vs psychological etiology of nonatopic food and chemical sensitivities, little systematic research has addressed the issues. The present study investigated self-reported illness from several common foods (wheat, dairy, eggs) and chemicals (pesticide, car exhaust, paint, perfume, new carpet), symptom patterns, and psychological profiles of a sample of young adult college students (n = 490, age 19.4 +/- 2.4, 52% female/48% male). Subjects were divided into 4 groups on the basis of sample medians for frequency of illness from the foods (FI) and chemicals (CI); high FI with high CI (FI/CI), high FI alone, high CI alone, and NOILL (low FI and CI). FI was associated with more defensiveness (denial of negativity) while CI was linked with more shyness (avoidance of novelty). Women outnumbered men in all groups (FI/CI: 61%; FI: 80% CI: 55%) except the NOILL (40% women). Nevertheless, the FI/CI, FI, and/or CI groups still had significantly higher total symptom scores as well as more indigestion, headache, and memory trouble than did the NOILL group, even after depression, anxiety, shyness, defensiveness, and gender were covaried. The illness groups reported significantly more limitation of foods that mobilize endogenous opioids or generate exogenous opioids (sweets, fats, bread) as well as more illness from opiate drugs, small amounts of beverage alcohol, and late meals. Nasal symptoms from pollens or animals were more common in the FI/CI (42%) and CI (42%) than in FI (26%) or NOILL (28%) groups. Premenstrual tension syndrome and irritable bowel were also more common in the FI/CI group. The findings indicate that young adults outside the clinical setting who are relatively higher in FI and/or CI have distinctive symptom and psychological patterns. Covariate analyses suggest that important symptoms in FI and CI individuals such as indigestion, headache, and memory problems may occur in addition to rather than as simply part of emotional distress. The data are consistent with a previously hypothesized role of olfactory-limbic and hypothalamic pathways and with a time-dependent sensitization model for illness from foods and chemicals.

Adolescent↗

A light- and developmentally-regulated DNA-binding interaction is common to the upstream sequences of the wheat Calvin cycle bisphosphatase genes.

We have characterised a DNA-binding interaction common to the upstream sequences of the wheat fructose-1,6-bisphosphatase (FBPase) and sedoheptulose-1,7-bisphosphatase (SBPase) genes. The recognition site for this sequence-specific binding activity, designated wheat FBPase factor (WF-1), is located within 125 bp of the transcription start site of each gene. Within these regions there are no sequence motifs similar to those shown to be important for light-regulated expression in other species. The binding activity was not detected in wheat root nuclear extracts, or in pea leaf extracts. There was a higher level of binding activity in light-grown than in dark-grown wheat leaves. The level was also found to decline when light-grown plants were given an extended dark treatment, but could be reinduced by light. Utilising the gradient of developmental maturity which exists within the wheat leaf it was found that WF-1 activity increases during leaf development.

Base Sequence↗

The native alpha 2 beta 2 tetramer is the only subunit structure of the insulin receptor in intact cells and purified receptor preparations.

The native subunit structure of the insulin receptor was reinvestigated by two-dimensional nonreducing/reducing gel electrophoresis. Human insulin receptor expressed in murine fibroblasts was found to be a single oligomer, the alpha 2 beta 2 heterotetramer. The structure was assessed using receptor metabolically labeled with [35S]methionine, and using receptor autophosphorylation at two levels of purification: the insulin affinity-purified receptor and the more commonly used wheat germ agglutinin-Sepharose-enriched fraction from whole membrane extracts. Lower molecular weight oligomers and free subunits were observed only upon heating the sample prior to electrophoresis. This artifact of sample handling was dependent upon three factors: (i) temperature, (ii) time of heating, and (iii) impurities typically present in partially purified receptor preparations. We conclude that the alpha 2 beta 2 tetramer is the only insulin receptor subunit structure native in intact cells and subsequently isolated from cell membranes.

Animals↗

Modeling light and temperature effects on leaf emergence in wheat and barley.

Phenological development affects canopy structure, radiation interception, and dry matter production; most crop simulation models therefore incorporate leaf emergence rate as a basic parameter. A recent study examined leaf emergence rate as a function of temperature and daylength among wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) cultivars. Leaf emergence rate and phyllochron were modeled as functions of temperature alone, daylength alone, and the interaction between temperature and daylength. The resulting equations contained an unwieldy number of constants. Here we simplify by reducing the constants by > 70%, and show leaf emergence rate as a single response surface with temperature and daylength. In addition, we incorporate the effect of photosynthetic photon flux into the model. Generic fits for wheat and barley show cultivar differences less than +/- 5% for wheat and less than +/- 10% for barley. Barley is more sensitive to daylength changes than wheat for common environmental values of daylength, which may be related to the difference in sensitivity to daylength between spring and winter cultivars. Differences in leaf emergence rate between cultivars can be incorporated into the model by means of a single, nondimensional factor for each cultivar.

Hordeum↗

[On antimutagenesis mechanism in plants].

The effect of nitrosylmethylurea (NMU) on the mitotic index and the frequency of cells with aberrations, as well as the effects of pre- and posttreatment with antioxidant ambiol on the NMU effects were studied on seedlings of common winter wheat Triticum aestivum, cultivar Moskovskaya 39. Both pre- and posttreatment with ambiol resulted in antimutagenic effect but after posttreatment, the effect was lower. Irrespective of type of seedling treatment with ambiol and the time of their fixation (45, 48, and 51 h), when mitotic index is plotted versus frequency of cells with aberrations, all experimental points fall on the same regression line with coefficient of correlation of -0.82 (P < 0.001). This implies that the same mechanism underlies antimutagenic effect irrespective of when the antimutagen was applied, before or after the knockout mutagen dose. This also suggests that the antimutagenic effect is independent of the degree of the mutagen-induced damage, because by the time of posttreatment, the volume of genome damage is already determined and the antimutagen fails to change it. Finally, this suggests that irrespective of time of antimutagen treatment, the mutation frequency is reduced by the mechanism of stimulated repopulation.

Antimutagenic Agents↗

Mycotoxin research in the Hungarian Central Veterinary Institute.

Mycotoxin research conducted in the Hungarian Central Veterinary Institute is reviewed briefly. The effect of zearalenone was studied by experiments in several animal species (cattle, pig, sheep, chicken, goose, duck, guinea fowl, fish) and in cell cultures in vitro. The chicken (Gallus domesticus) has proved to be resistant to the toxin. In the susceptible species zearalenone causes the most severe damage to the sexual organs. The metabolism of zearalenone, T-2 toxin and diacetoxyscirpenol was also studied. These toxins are resistant to physicochemical factors but are easily transformed in biologically active environment. The acute toxicity study of trichothecene toxins is described along with morphological changes, with particular respect to those of the lymphoid and myeloid organs. Trichothecene toxins impair the natural defence mechanism of the organism and result in the manifestation of different diseases (swine dysentery, caecal coccidiosis). Deoxynivalenol contamination of the feeds, first of all of wheat, is common in Hungary. Its effect on pig fattening was investigated in a field trial. The toxins of different storehouse moulds were also examined.

Animals↗

Characterization of a Gibberellin-Insensitive Dwarf Wheat, D6899 : EVIDENCE FOR A REGULATORY STEP COMMON TO MANY DIVERSE RESPONSES TO GIBBERELLINS.

The effects of gibberellin in two wheat varieties, Nainari 60 and D6899, have been studied. D6899 is a dwarf wheat having a single locus mutation, the Rht 3 gene ("Tom Thumb" gene), which is located on chromosome 4A. When compared with the standard height wheat variety, Nainari 60, D6899 does not have a slowdown in cellular metabolism such as respiration rate, protein content, rate of protein synthesis, and uptake of amino acids. The content of ATP is even higher in D6899. However, all of the gibberellin-mediated physiological processes that we have studied, including leaf elongation, synthesis and release of hydrolytic enzymes, and secretion of phosphate ions and reducing sugars in aleurone layers, are retarded in D6899. D6899 and Nainari 60 have essentially the same uptake and metabolism of gibberellin and their levels of endogenous inhibitors such as abscisic acid do not differ drastically. The dosage-response curve of the gibberellin-mediated alpha-amylase production indicates that a rate-limiting step, which is common to many of the diverse gibberellin responses, is partially blocked in D6899 wheat.

Journal Article↗

Haplotype diversity at fusarium head blight resistance QTLs in wheat.

Fusarium head blight (FHB) reduces grain yield and quality in common and durum wheat. Host FHB resistance is an effective control measure that is achieved by stacking multiple resistance genes into a wheat line. Therefore, breeders would benefit from knowing which resistance sources carry different resistance genes. A diverse collection of FHB-resistant and -susceptible wheat lines was characterized with microsatellite markers linked to FHB resistance quantitative trait loci (QTLs) on chromosomes 2DL, 3BS (distal to the centromere), 3BSc (proximal to the centromere), 4B, 5AS and 6BS identified in wheat lines Maringa, Sumai 3 and Wuhan 1. Putative Sumai 3 QTLs were commonly observed in advanced breeding lines, whereas putative Maringa and Wuhan 1 QTLs were relatively rare. Marker data suggested the 3BS, 3BSc and 5AS QTLs in the Brazilian cv. Maringa were derived from Asian germplasm and not from Frontana or other Brazilian lines. Haplotype diversity was reduced near the 5AS QTL, which might impact the deployment of this QTL. Finally, Brazilian germplasm was not closely related to other resistance sources and might be useful for pyramiding with Asian wheat-derived FHB resistance.

Agriculture↗

Enhancing the resistance of triticale by using genes from wheat and rye.

At present two separate nomenclature systems exist for wheat and rye. This paper provides a proposed common catalogue of wheat, rye and triticale resistance gene symbols. More than 130 postulated wheat resistance genes are listed. Over 39 rye and 6 triticale resistance (R) genes have been identified and named. Genes responsible for reaction to powdery mildew and to leaf, stem and yellow rusts are the best-represented group of resistance genes. From the common catalogue it can be concluded that there exists a potential for further transfer of rye resistance genes to wheat and triticale. Many molecular markers can be applied for marker-assisted gene transfer, but the expression of the R genes in the new genetic background of triticale remains to be investigated.

Edible Grain↗

High transferability of bread wheat EST-derived SSRs to other cereals.

The increasing availability of expressed sequence tags (ESTs) in wheat (Triticum aestivum) and related cereals provides a valuable resource of non-anonymous DNA molecular markers. In this study, 300 primer pairs were designed from 265 wheat ESTs that contain microsatellites in order to develop new markers for wheat. Their level of transferability in eight related species [Triticum durum, T. monococcum, Aegilops speltoides, Ae. tauschii, rye (Secale cereale), barley (Hordeum vulgare), Agropyron elongatum and rice (Oryza sativa)] was assessed. In total, 240 primer pairs (80%) gave an amplification product on wheat, and 177 were assigned to wheat chromosomes using aneuploid lines. Transferability to closely related Triticeae species ranged from 76.7% for Ae. tauschii to 90.4% for T. durum and was lower for more distant relatives such as barley (50.4%) or rice (28.3%). No clear putative function could be assigned to the genes from which the simple sequence repeats (SSRs) were developed, even though most of them were located inside ORFs. BLAST: analysis of the EST sequences against the 12 rice pseudo-molecules showed that the EST-SSRs are mainly located in the telomeric regions and that the wheat ESTs have the highest similarity to genes on rice chromosomes 2, 3 and 5. Interestingly, most of the SSRs giving an amplification product on barley or rice had a repeated motif similar to the one found in wheat, suggesting a common ancestral origin. Our results indicate that wheat EST-SSRs show a high level of transferability across distantly related species, thereby providing additional markers for comparative mapping and for following gene introgressions from wild species and carrying out evolutionary studies.

Chromosome Mapping↗

Population structure within lineages of Wheat streak mosaic virus derived from a common founding event exhibits stochastic variation inconsistent with the deterministic quasi-species model.

Structure of Wheat streak mosaic virus (WSMV) populations derived from a common founding event and subjected to serial passage at high multiplicity of infection (MOI) was evaluated. The founding population was generated by limiting dilution inoculation. Lineages of known pedigree were sampled at passage 9 (two populations) and at passage 15, with (three populations) or without mixing (four populations) of lineages at passage 10. Polymorphism within each population was assessed by sequencing 17-21 clones containing a 1371 nt region (WSMV-Sidney 81 nts 8001-9371) encompassing the entire coat protein cistron and flanking regions. Mutation frequency averaged approximately 5.0 x 10(-4)/nt across all populations and ranged from 2.4 to 11.6 x 10(-4)/nt within populations, but did not consistently increase or decrease with the number of passages removed from the founding population. Shared substitutions (19 nonsynonymous, 10 synonymous, and 3 noncoding) occurred at 32 sites among 44 haplotypes. Only four substitutions became fixed (frequency = 100%) within a population and nearly one third (10/32) never achieved a frequency of 10% or greater in any sampled population. Shared substitutions were randomly distributed with respect to genome position, with transitions outnumbering transversions 5.4:1 and a clear bias for A to G and U to C substitutions. Haplotype composition of each population was unique with complexity of each population varying unpredictably, in that the number and frequency of haplotypes within a lineage were not correlated with number of passages removed from the founding population or whether the population was derived from a single or mixed lineage. The simplest explanation is that plant virus lineages, even those propagated at high MOI, are subject to frequent, narrow genetic bottlenecks during systemic movement that result in low effective population size and stochastic changes in population structure upon serial passage.

Base Sequence↗

[Anaphylactic reactions after intake of products with wheat].

BACKGROUND: Food allergy is common in small children. Sensitisation to wheat is frequent, reactions to ingested wheat more seldom and mostly associated with atopic eczema and exercise-induced anaphylaxis. MATERIAL AND METHODS: Eleven children with severe reactions after ingestion of small amounts of wheat were referred to us for further diagnostic procedures. Skin prick tests, total and specific IgE as well as oral provocation test were performed. RESULTS: All but two children had strongly positive skin prick tests results to wheat. The same two children had specific IgE to Immunocap f4 (a-amylase) < 10 kUa/L while the nine other children had specific IgE to f4 > 29 kUa/L. All children had general reactions when provoked orally with small amounts of wheat with the need for administration of adrenalin and in most cases oral steroids. INTERPRETATION: It is important to identify children at risk for general reactions to wheat, as cereals are staple foods in our community. Identification of the proteins involved in serious reactions is essential.

Anaphylaxis↗

High-resolution mapping of the leaf rust disease resistance gene Lr1 in wheat and characterization of BAC clones from the Lr1 locus.

Leaf rust is the most common disease in wheat production. There are more than 45 specific resistance genes described and used in wheat breeding to control epidemics of leaf rust, but none of them has been cloned. The leaf rust disease resistance gene 1 ( Lr1) is a good model gene for isolation by map-based cloning because it is a single, dominant gene which is located in the distal region of chromosome 5DL of wheat. As the first step towards the isolation of this gene we constructed a high-resolution genetic map in the region of the Lr1 locus by saturation mapping of two large segregating F(2) populations (Thatcher Lr1 x Thatcher, Thatcher Lr1 x Frisal). The resistance gene Lr1 was delimited in a 0.16-cM region between the RFLP markers ABC718 and PSR567 (0.12 cM from ABC718 and 0.04 cM from PSR567). A genomic BAC library of Aegilops tauschii (D genome) was screened using the RFLP markers ABC718 and PSR567. Five positive BAC clones were identified by ABC718 and four clones by PSR567. Two NBS-LRR type of resistance gene analogs, which encode proteins highly homologous to the bacterial blight disease resistance protein Xa1 of rice, were identified on BAC clones isolated with PSR567. Polymorphic BAC end probes were isolated from both ends of a 105-kb large BAC clone identified by ABC718. The end probes were mapped at the same locus as ABC718, and no recombination event was found within 105 kb around ABC718 in our analysis of more than 4,000 gametes.

Amino Acid Sequence↗

Monitoring and identification of aflatoxins in wheat, gram and maize flours in Bihar state (India).

The occurrence of aflatoxins in some common food commodities: wheat, gram and maize flours is reported. Of the total 416 collections during three years (1987-1989) 162 were recorded to be aflatoxin-positive. The incidence of aflatoxins was maximum in the samples of 1987. Most of the contaminated samples contained aflatoxins at levels above 20 micrograms/kg. The environmental conditions and traditional agronomic and storage practices relating to aflatoxin incidence are discussed.

Aflatoxins↗