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The relationship between high-molecular-weight glutenin subunit composition and the quality of Japanese hexaploid wheat lines.

To reveal the high-molecular-weight (1-1MW) glutenin subunit composition, the seed storage proteins of 40 Japanese wheat (Triticum aestivum) lines were fractionated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis to determine their HMW glutenin subunit composition. These were identified by comparison of subunit mobility with that previously found in hexaploid wheat. Twelve different, major glutenin HMW subunits were identified. Each line contained three to five subunits, and 11 different glutenin subunit patterns were observed for 11 alleles in Japanese lines. The Glu-1 quality scores were not particularly high for most of the Japanese wheats in the southern part of Japan (Kyushu district). However, the Glu-1 quality scores of several wheat lines in the Hokkaido area (north Japan) were high. South Japanese wheat lines showed specialty allelic variation in the glutenin HMW 145 kfla subunit, different from those in non-Japanese hexaploid wheats.

Alleles↗

Regulation by biotic and abiotic stress of a wheat germin gene encoding oxalate oxidase, a H2O2-producing enzyme.

Germins and germin-like proteins (GLPs) constitute a ubiquitous family of plant proteins that seem to be involved in many developmental and stress-related processes. Wheat germin has been extensively studied at the biochemical level: it is found in the apoplast and the cytoplasm of germinating embryo cells and it has oxalate oxidase activity (EC 1.2.3.4). Germin synthesis can also be induced in adult wheat leaves by auxins and by a fungal pathogen but it remains to be determined whether the same gene is involved in developmental, hormonal and stress response. In this work, we have studied the expression of one of the wheat germin genes, named gf-2.8, in wheat as well as in transgenic tobacco plants transformed with either this intact gene or constructs with GUS driven by its promoter. This has allowed us to demonstrate that expression of this single gene is both developmentally and pathogen-regulated. In addition, we show that expression of the wheat gf-2.8 germin gene is also stimulated by some abiotic stresses, especially the heavy metal ions Cd2+, Cu2+ and Co2+. Several chemicals involved in stress signal transduction pathways were also tested: only polyamines were shown to stimulate expression of this gene. Because regulation of the wheat gf-2.8 germin gene is complex and because its product results in developmental and stress-related release of hydrogen peroxide in the apoplast, it is likely that it plays an important role in several aspects of plant growth and defence mechanisms.

Antioxidants↗

A survey on the occurrence of mycotoxins in wheat and maize from western Romania.

Samples of wheat (n = 25) and maize (n = 30) for animal consumption, collected in 1997 after harvest from western Romania, were analyzed by enzyme immunoassays for mycotoxin contamination. Toxins analyses included deoxynivalenol (DON), 3-acetylDON, 15- acetylDON, fusarenone X (FX), T-2 Toxin (T-2), diacetoxyscirpenol (DAS), zearalenone (ZEA), fumonisin B(1) (FB(1)), aflatoxin B(1) (AFB(1)), ochratoxin A (OA), and citrinin (CT). DON and acetylDONs were the major contaminants in wheat (100%) and maize (46%). Median values for DON, 3-acetylDON, and 15-acetylDON were 880 microg kg(-1), 66 microg kg(-1), and 150 microg kg(-1) in wheat, and 890 microg kg(-1), 180 microg kg(-1), and 620 microg kg(-1) in maize, respectively. Additionally, 3,15-diacetylDON was detected in some samples by HPLC-EIA analysis. All samples were negative for FX (<150 microg kg(-1)). T-2 was found in wheat (n = 6) and maize (n = 1) at levels between 13 and 63 microg kg(-1). DAS (2.6 microg kg(-1)) was found in one maize sample. ZEA occurred in all wheat and in four maize samples, median values were 10 microg kg(-1) and 250 microg kg(-1), respectively. One maize sample contained FB(1) (140 microg kg(-1)). All samples were AFB(1)-negative (<4 microg kg(-1)). OA was found in one wheat sample (37 microg kg(-1)), CT was found in one maize sample (580 microg kg(-1)). This first reported natural occurrence of a range of mycotoxins in Romanian feeding stuff shows that DON and acetyl DONs may be present at levels which may affect animal production.

Journal Article↗

DNA binding mediated by the wheat HMGa protein: a novel instance of selectivity against alternating GC sequence.

The high-mobility-group (HMG) chromosomal protein wheat HMGa was purified to homogeneity and tested for its binding characteristics to double-stranded DNA. Wheat HMGa was able to bind to P268, an A/T-rich fragment derived from the pea plastocyanin gene promoter, producing a small mobility shift in gel retardation assays where the bound complex was sensitive to addition of proteinase K but resistant to heat treatment of the protein, consistent with the identity of wheat HMGa as a putative HMG-I/Y protein. Gel retardation assays and southwestern hybridization analysis revealed that wheat HMGa could selectively interact with the DNA polynucleotides poly(dA).poly(dT), poly(dAdT).poly(dAdT), and poly(dG).poly(dC), but not with poly(dGdC).poly(dGdC). Surface plasmon resonance analysis determined the kinetic and affinity constants of sensor chip-immobilized wheat HMGa for double-stranded DNA 10-mers, revealing a good affinity of the protein for various dinucleotide combinations, except that of alternating GC sequence. Thus contrary to prior reports of a selectivity of wheat HMGa for A/T-rich DNA, the protein appears to be able to interact with sequences containing guanine and cytosine residues as well, except where G/C residues alternate directly in the primary sequence.

Base Sequence↗

Puroindolines: the molecular genetic basis of wheat grain hardness.

The variation in grain hardness is the single most important trait that determines end-use quality of wheat. Grain texture classification is based primarily on either the resistance of kernels to crushing or the particle size distribution of ground grain or flour. Recently, the molecular genetic basis of grain hardness has become known, and it is the focus of this review. The puroindoline proteins a and b form the molecular basis of wheat grain hardness or texture. When both puroindolines are in their 'functional' wild state, grain texture is soft. When either one of the puroindolines is absent or altered by mutation, then the result is hard texture. In the case of durum wheat which lacks puroindolines, the texture is very hard. Puroindolines represent the molecular-genetic basis of the Hardness locus on chromosome 5DS and the soft (Ha) and hard (ha) alleles present in hexaploid bread wheat varieties. To date, seven discrete hardness alleles have been described for wheat. All involve puroindoline a or b and have been designated Pina-D1b and Pinb-D1b through Pinb-D1g. A direct role of a related protein, grain softness protein (as currently defined), in wheat grain texture has yet to be demonstrated.

Alleles↗

Sequences of the waxy loci of wheat: utility in analysis of waxy proteins and developing molecular markers.

The waxy proteins from a number of genetic backgrounds of wheat and its progenitors were analyzed by SDS-PAGE. The deduced amino acid sequences of the waxy proteins of three diploid progenitor species indicated several key amino acid substitutions, which could explain the differences observed in the electrophoretic mobilities of the wheat waxy proteins. A slight difference observed in the apparent molecular weight of the WX-A1 protein of diploid and polyploid wheat was explained by amino acid substitutions or variations in predicted protein structures. Further, twelve different partial genomic clones, representing the individual waxy loci of the various diploid, tetraploid and hexaploid wheats, were isolated and compared. The results indicated significant variations in intron 4 and led to identification of sequences unique to the individual waxy genes and genomes of wheat and its proposed progenitors. The sequence variations observed have a great potential for development as molecular markers for identification of specific waxy loci and study of the various waxy mutants of wheat.

Base Sequence↗

Optimum nitrogen use and reduced nitrogen loss for production of rice and wheat in the Yangtse Delta region.

A long-term field and lysimeter experiment under different amount of fertilizer-N application was conducted to explore the optimal N application rates for a high productive rice-wheat system and less N leaching loss in the Yangtse Delta region. In this region excessive applications of N fertilizer for the rice-wheat production has resulted in reduced N recovery rates and environment pollution. Initial results of the field experiments showed that the optimal N application rate increased with the yield. On the two major paddy soils (Hydromorphic paddy soil and Gleyed paddy soil) of the region, the optimal N application rate was 225-270 kg N hm(-2) for rice and 180-225 kg N hm(-2) for wheat, separately. This has resulted in the highest number of effective ears and Spikelets per unit area, and hence high yield. Nitrogen leaching in the form of NO(3-)-N occurs mainly in the wheat-growing season and in the ponding and seedling periods of the paddy field. Its concentration in the leachate increased with the N application rate in the lysimeter experiment. When the application rate reached 225 kg N hm(-2), the concentration rose to 5.4-21.3 mgN l(-1) in the leachate during the wheat-growing season. About 60% of the leachate samples determined contained NO(3-)-N beyond the criterion (NO(3-)-N 10 mg l(-1)) for N pollution. In the field experiment, when the N application rate was in the range of 270-315 kg hm(-2), the NO(3-)-N concentration in the leachate during the wheat-growing season ranged from 1.9 to 11.0 mg l(-1). About 20% of the leachate samples reached close to, and 10% exceeded, the criterion for N pollution. Long-term accumulation of NO(3-)-N from leaching will no doubt constitute a potential risk of N contamination of the groundwater in the Yangtse Delta Region.

Agriculture↗

Extrusion of wheat or sorghum and/or addition of exogenous enzymes to pig diets influences the large intestinal microbiota but does not prevent development of swine dysentery following experimental challenge.

A study was made of dietary influences on the large intestinal microbiota of pigs and on the incidence of swine dysentery (SD) after experimental infection with Brachyspira hyodysenteriae, the aetiological agent of SD. Animals were fed diets based either on wheat (expts 1 and 2) or sorghum (expt 2). Grains were ground and fed either raw or after high temperature and pressure extrusion and/or after addition of exogenous enzymes to the whole diet to reduce the starch and soluble non-starch polysaccharides available for fermentation in the large intestine. Limiting fermentation creates conditions that apparently reduce the incidence of SD after infection with B. hyodysenteriae. The diets were fed to weaned pigs for 4-6 weeks, then half the animals on each diet were killed and gut samples collected for microbiology. The treatments had little effect on bacterial numbers. In expt 1, dietary extrusion of wheat reduced lactobacilli in the large intestine. Addition of enzymes to extruded wheat-based diets in expt 2 reduced facultative anaerobes and increased non-sporing anaerobes. Addition of enzymes to a raw sorghum diet in expt 3 decreased numbers of facultative anaerobes, while extrusion of sorghum increased total anaerobes. Bacteroides spp. and Fusobacterium spp., which act in synergy with B. hyodysenteriae in SD, were isolated at a higher percentage in pigs fed the untreated wheat diet than in pigs fed the treated wheat diets. Following experimental infection the incidence of SD amongst pigs fed treated wheat diets was slightly lower than those fed the untreated diet, but with sorghum-based diets the opposite was found. Overall, the different dietary treatments used did not significantly reduce SD.

Animals↗

Molecular cloning of a new receptor-like kinase gene encoded at the Lr10 disease resistance locus of wheat.

More than 100 resistance genes against wheat rust pathogens have been described in wheat and its relatives. Although many of them have been extensively used in wheat resistance breeding, none of these resistance loci has yet been analyzed at the molecular level. By screening a set of near-isogenic lines carrying different leaf rust resistance genes with a wheat probe encoding a serine/ threonine protein kinase, we detected a polymorphic DNA fragment in the line with the Lr10 resistance gene. This fragment mapped to the Lr10 disease resistance locus and encodes a receptor-like protein kinase which we called LRK10. LRK10 contains a new type of extracellular domain not found in known plant or animal receptor kinases. Several conserved amino acids in S-domain glycoproteins and receptor-like kinases were also found in LRK10, suggesting that LRK10 and S-domain proteins belong to the same superfamily of specific recognition molecules in plants. Lrk10 was expressed at low levels in young seedlings and belongs to a gene family. Analysis of wheat lines with and without the Lr10 gene demonstrated that Lrk10 and Lr10 belong to the same genetic locus. We conclude that gene isolation based on protein kinase homology can identify new receptor domains and provide candidates for disease resistance genes in the complex wheat genome.

Alleles↗

Comparison of wheat and rye flour skin prick test solutions for diagnosis of baker's asthma.

BACKGROUND: Skin prick tests (SPTs) play an important role in the diagnosis of baker's asthma and in the investigation of sensitization frequencies in field studies. It was the aim of our study to compare different SPT solutions for wheat and rye flour sensitization and to assess the validity of test results. METHODS: Skin prick tests with wheat and rye flour were performed in parallel with extracts from different companies and compared with the results of bronchial challenge tests with both flours (69 rye flour and 51 wheat flour challenge tests). Additionally, specific immunoglobulin E (sIgE) to wheat and rye flour were tested. SPT solutions were analysed for protein content and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: Skin prick test solutions for diagnosis of wheat and rye flour sensitization from three companies differed in protein concentrations and composition with the consequence of widely differing SPT results. Sensitivity of SPTs in comparison with allergen-specific bronchial challenge as a gold standard was between 40 and 67%, specificity was between 86 and 100%, the positive predictive value (PPV) ranged from 81 to 100% and the negative predictive value (NPV) from 44 to 70%. These numbers were only marginally affected by using a combination of challenge test result and sIgE value as a more specific gold standard. CONCLUSION: Improvement and standardization of SPT extracts for wheat and rye flour is highly recommended.

Adult↗

Characterization of a gene from chromosome 1B encoding the large subunit of ADPglucose pyrophosphorylase from wheat: evolutionary divergence and differential expression of Agp2 genes between leaves and developing endosperm.

A full-length genomic clone containing the gene encoding the large subunit of the ADPglucose pyrophosphorylase (Agp2), was isolated from a genomic library prepared from etiolated shoots of hexaploid wheat (Triticum aestivum L., cv, Chinese Spring). The coding region of this gene is identical to one of the cDNA clones previously isolated from a developing wheat grain cDNA library and is therefore an actively transcribed gene. The sequence represented by the cDNA spans 4.8 kb of the genomic clone and contains 15 introns. 2852 bp of DNA flanking the transcription start site of the gene was cloned upstream of the GUS (beta-glucuronidase) reporter gene. This Agp2::GUS construct and promoter deletions were used to study the pattern of reporter gene expression in both transgenic tobacco and wheat plants. Histochemical analysis of GUS expression in transgenic tobacco demonstrated that the reporter gene was expressed in guard cells of leaves and throughout the seed. In transgenic wheat, reporter gene expression was confined to the endosperm and aleurone with no expression in leaves. The cloned Agp2 gene was located to chromosome 1B by gene-specific PCR with nullisomic-tetrasomic lines. Northern analysis demonstrated that the Agp2 genes are differentially expressed in leaves and developing endosperm; while all three classes of Agp2 genes are transcribed in developing wheat grain endosperm, only one is transcribed in leaves. The differences between the Agp2 genes are discussed in relation to the evolution of hexaploid wheat.

Journal Article↗

Effects of wheat bran on exocrine pancreas secretion in the pig.

The aim of this study was to determine the effect of wheat bran consumption on exocrine pancreas secretion in pigs. Sixteen Large-White pigs were divided into two groups. The first group (control) was fed a diet without wheat bran and the second one (experimental) a diet containing 40% wheat bran. After one week the animals were fitted with two permanent fistulae (in the pancreatic duct and the duodenum) and/or with a catheter in a carotid artery. After an 8-day recovery period, pancreatic secretion (volume, protein content and output, chymotrypsin, trypsin, lipase and amylase activities) and plasma levels of some gastro-intestinal peptides [secretin, cholecystokinin (CCK), vasoactive intestinal peptide (VIP), somatostatin and pancreatic polypeptide (PP)] were measured over an experimental period of 5 days. The results show that wheat bran intake induced an increase in the volume (+ 115%) and protein output (+ 36%) of the pancreatic juice secreted in a 24-hour period, whereas protein concentration decreased. All enzyme activities were enhanced by wheat bran. The plasma levels of secretin, VIP, somatostatin and PP were higher in the experimental than in the control group. On the contrary, plasma CCK levels were not affected by wheat bran consumption.

Animals↗

The wheat and barley vernalization gene VRN3 is an orthologue of FT.

Winter wheat and barley varieties require an extended exposure to low temperatures to accelerate flowering (vernalization), whereas spring varieties do not have this requirement. In this study, we show that in these species, the vernalization gene VRN3 is linked completely to a gene similar to Arabidopsis FLOWERING LOCUS T (FT). FT induction in the leaves results in a transmissible signal that promotes flowering. Transcript levels of the barley and wheat orthologues, designated as HvFT and TaFT, respectively, are significantly higher in plants homozygous for the dominant Vrn3 alleles (early flowering) than in plants homozygous for the recessive vrn3 alleles (late flowering). In wheat, the dominant Vrn3 allele is associated with the insertion of a retroelement in the TaFT promoter, whereas in barley, mutations in the HvFT first intron differentiate plants with dominant and recessive VRN3 alleles. Winter wheat plants transformed with the TaFT allele carrying the promoter retroelement insertion flowered significantly earlier than nontransgenic plants, supporting the identity between TaFT and VRN-B3. Statistical analyses of flowering times confirmed the presence of significant interactions between vernalization and FT allelic classes in both wheat and barley (P < 0.0001). These interactions were supported further by the observed up-regulation of HvFT transcript levels by vernalization in barley winter plants (P = 0.002). These results confirmed that the wheat and barley FT genes are responsible for natural allelic variation in vernalization requirement, providing additional sources of adaptive diversity to these economically important crops.

Adaptation, Physiological↗

Genes encoding plastid acetyl-CoA carboxylase and 3-phosphoglycerate kinase of the Triticum/Aegilops complex and the evolutionary history of polyploid wheat.

The classic wheat evolutionary history is one of adaptive radiation of the diploid Triticum/Aegilops species (A, S, D), genome convergence and divergence of the tetraploid (Triticum turgidum AABB, and Triticum timopheevii AAGG) and hexaploid (Triticum aestivum, AABBDD) species. We analyzed Acc-1 (plastid acetyl-CoA carboxylase) and Pgk-1 (plastid 3-phosphoglycerate kinase) genes to determine phylogenetic relationships among Triticum and Aegilops species of the wheat lineage and to establish the timeline of wheat evolution based on gene sequence comparisons. Triticum urartu was confirmed as the A genome donor of tetraploid and hexaploid wheat. The A genome of polyploid wheat diverged from T. urartu less than half a million years ago (MYA), indicating a relatively recent origin of polyploid wheat. The D genome sequences of T. aestivum and Aegilops tauschii are identical, confirming that T. aestivum arose from hybridization of T. turgidum and Ae. tauschii only 8,000 years ago. The diploid Triticum and Aegilops progenitors of the A, B, D, G, and S genomes all radiated 2.5-4.5 MYA. Our data suggest that the Acc-1 and Pgk-1 loci have different histories in different lineages, indicating genome mosaicity and significant intraspecific differentiation. Some loci of the S genome of Aegilops speltoides and the G genome of T. timophevii are closely related, suggesting the same origin of some parts of their genomes. None of the Aegilops genomes analyzed is a close relative of the B genome, so the diploid progenitor of the B genome remains unknown.

Acetyl-CoA Carboxylase↗

Genetic control of endosperm amylase activity and gibberellic Acid responses in standard-height and short-statured wheats.

In contrast to standard-height wheat genotypes, short-statured wheats having major genes for dwarfness do not show increased seedling growth after treatment with gibberellic acid. Endogenous gibberellic acid induces synthesis of amylase in the endosperm of germinating seeds, but the amount of amylase synthesized is greatly increased by exogenous gibberellic acid treatment in standard-height and in short-statured wheats that have dwarfing genes from the variety "Norin 10." "D6899," which has the "Tom Thumb" gene for height reduction, had about one-fourth of the amylase activity of standard-height and Norin 10-derived, short-statured wheats. This genotype showed little or no increased amylase activity after gibberellic acid treatment. Genetic analyses showed that the amount of amylase synthesized was controlled by a single gene and was dependent on the number of copies of the structural gene present in the endosperm. Dwarfism in wheat may be related to a blockage in gibberellic acid utilization because other workers have found that the amount of endogenous amylase synthesized in Norin 10-derived, short-statured wheats is not growth-limiting, but it is not known if low amylase synthesis is related to dwarfism in the Tom Thumb derivative. No recombinants were recovered in a small population, suggesting that the Tom Thumb gene may pleiotropically affect plant height and the lack of response to gibberellic acid in amylase synthesis and seedling growth.

Journal Article↗

Fusarium graminearum and deoxynivalenol contamination in the durum wheat area of Argentina.

Fusarium graminearum head blight of wheat is a destructive disease of the world's wheat-growing areas. This work was performed to analyze the distribution and contamination of deoxynivalenol (DON) and its relationship with F. graminearum kernel invasion in Argentina durum wheat area during two consecutive harvests. A total of 147 samples (cultivars and lines) of durum wheat from 5 locations of the major cropping area (Southern Buenos Aires Province) were analyzed. Percentage of F. graminearum kernel infection was evaluated following the blotter test (ISTA method) and fusarotoxins were analyzed by thin layer chromatography. None of the varieties and lines were free of F. graminearum infection. In the first harvest fungal invasion was very low. From 40 samples, 55% showed DON contamination but only 4 samples (10%) were higher than 2 ppm. In the second harvest, a crop year conducive to scab development, the highest level of F. graminearum kernel invasion observed was 42% on a sample from the humid area (eastern Buenos Aires Province) DON was detected in 47 (78.2%) of 60 samples analyzed and 19 (31.6%) showed levels of DON higher than those established in the guidelines in Canada and USA for food and feedstuff. In both years all locations situated in the humid area showed levels ranging from 0 to > 8 ppm. Within the durum wheat area differences among locations were found. This analysis indicates the need for more information on the problem and distribution of Fusarium mycotoxins in durum wheat grown in Argentina.

Argentina↗

Seedling and adult plant resistance to Sitobion avenae (Hemiptera: Aphididae) in Triticum monococcum (Poaceae), an ancestor of wheat.

Cereal aphids are important pests of wheat, Triticum aestivum L. and Triticum durum Desf. Crop resistance is a desirable method for managing cereal aphids in central North America, where the dominant crop, spring-sown wheat, has a low value per unit area. A diploid ancestor of wheat, Triticum monococcum L., is reported to be partially resistant to Sitobion avenae (Fabricius), the most damaging cereal aphid in the region. To identify potential sources of resistance, 42 accessions of T. monococcum and three cultivated wheats were infested with aphids, seedlings for six days and adult plants for 21 days. Overall resistance was estimated by the biomass loss of foliage and spikes in relation to uninfested control plants. Antibiosis was estimated by the gain in biomass of aphids during infestation, and tolerance was estimated as a biomass conversion ratio, overall resistance divided by antibiosis. A few T. monococcum accessions exhibited partial resistance. No relationship was found between seedling and adult plant resistance: the former exhibited primarily antibiosis and the latter primarily tolerance. Two accessions with antibiosis reduced aphid biomass by 60% compared with commercial wheats. Tolerance was correlated with growth potential, and was useful only in accessions with high growth potential. Four accessions exhibited tolerance levels at least 30% greater than commercial wheats. Highly susceptible accessions also were identified, which would be useful for investigating the inheritance of antibiosis and tolerance.

Animals↗

The effects on chicks of dietary fibre from different sources: a growth factor in wheat bran.

1. Groups of chicks were given a low-residue diet with or without supplements of dietary fibre in the form of wheat bran, wheat straw or bagasse. Growth and food conversion efficiency (g weight gained/g food eaten; FCE) during the first 4 weeks of life were measured. 2. In every one of seven experiments supplementation of the diet with 100 g wheat bran/kg resulted in improved growth, and in three experiments FCE was also increased. 3. Supplementation with coarsely-milled wheat straw to provide an amount of unavailable carbohydrate equivalent to that in the bran diet resulted in poorer growth; finely-milled wheat straw had little effect on growth. 4. The growth-promoting effect of bran was destroyed by sterilization with heat or gamma-radiation. 5. In some experiments weights, lengths and volumes of small intestines were measured. Differences in intestinal dimensions between birds given the diet with and without fibre were not consistent, nor were they correlated with growth rate or FCE. 6. Histometric observations on small intestines from a few birds indicated that those given coarse wheat straw had longer vili and thicker muscularis layers, and the caecal tonsils had a greater area of lymphoid tissue and more follicles.

Animals↗