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Characterisation of mildew resistant wheat-rye substitution lines and identification of an inverted chromosome by fluorescent in situ hybridisation.

Seven different mildew resistant wheat lines derived from crosses between triticale and bread wheat were examined by molecular cytogenetics and chromosome C-banding in order to determine their chromosomal composition. Genomic in situ hybridisation (GISH) showed the presence of rye germplasm in all the lines and identified three substitution lines, three double substitution lines and one addition-substitution line. C-banding identified rye chromosomes 1R and 4R in the addition-substitution line, rye chromosomes 1R and 6R in two substitution lines and 1R and 2R in the third line, and rye chromosome 1R in the three substitution lines. Two of the latter lines (7-102 and 7-169) contained a modified form of the chromosome; fluorescent in situ hybridisation (FISH) using five different repetitive DNA-probes showed a pericentric inversion of 1R in both lines. The breakpoints of the 1R inversion were between (1) the 5S rDNA site and the NOR-region on the satellite of the short arm, and (2) between two AAC(5) sites close to the centromere on the long arm. The role of the rye chromosomes in the mildew resistance, the utilisation of the inverted 1R and the significance of the lines in wheat breeding are discussed.

Chromosome Banding↗

Utilisation of rye by chickens: effect of microbial status, diet gamma irradiation and sodium taurocholate supplementation.

Several experiments were conducted to determine the influence of intestinal microflora when rye diets are fed to young chicks. In a conventional environment dietary rye severely depressed growth, amino acid and fat retention, and metatarsal bone ash. These conditions could be effectively alleviated by gamma irradiation of the diet and maintenance in a germ-free environment. In gnotobiotic chicks associated with streptococci dietary rye caused effects similar to those seen in a conventional environment, however the responses were less severe. Since addition of sodium taurocholate improved fat retention from a rye diet to a greater extent than from a wheat diet, a deficiency of conjugated bile salts might contribute to the steatorrhea observed in rye-fed chicks.

Amino Acids↗

Occurrence of ochratoxin A in Danish wheat and rye, 1992-99.

Ochratoxin A concentrations in rye and wheat in Denmark for 1992-99 are reported. The results show that the concentration of ochratoxin A is higher in rye than in wheat for both conventionally and organically grown rye and wheat. The levels in organically grown rye are higher than in conventionally grown based on multiyear mean contents. However, the difference between the two groups of cereals has decreased since the Danish food-monitoring system for ochratoxin A was started in 1986; 2.0% of all samples exceeded the Danish maximum limit of 5 micro g kg(-1) introduced in 1995. For rye samples, 3.2% exceeded the maximum limit, and for wheat samples, 0.5% exceeded the maximum limit.

Carcinogens↗

The apparent ileal and faecal digestibilities of amino acids and carbohydrates of rye, barley and triticum grains in pigs.

Six pigs of initial weight 40 +/- 1.7 kg with a T-piece cannula fitted at the terminal ileum were given diets composed of 96.3% of rye, barley or triticum grains ground to pass the 2.6 mm mesh, 3.3% mineral and vitamin mixture and 0.4% Cr2O3. Each diet was given to all pigs in a Latin square design, and faeces and ileal digesta were collected on 7, 8, 9 and 10, 11 and 12 day of feeding each diet, respectively. The grains varied widely in the content of pentosans, polyuroids, beta-glucan and monosaccharides. The antitrypsin activities were 0.84, 2.73 and 1.62 TUI/g in barley, rye and triticum, respectively. Ileal and faecal apparent digestibilities of nitrogen and amino acids were higher (P less than or equal to 0.01) in Triticum than in barley and rye. The digestibility of carbohydrates (pentosans, beta-glucan, polyuroids and non-structural carbohydrates) were lower in pigs fed a rye diet than of those fed a Triticum diet. The results are discussed with respect of the role to non starch polysaccharides, especially pentosans in digestibility of nitrogen and energy in animals receiving a rye diet.

Amino Acids↗

The rye mutants identify a role for Ssn/Srb proteins of the RNA polymerase II holoenzyme during stationary phase entry in Saccharomyces cerevisiae.

Saccharomyces cerevisiae cells enter into a distinct resting state, known as stationary phase, in response to specific types of nutrient deprivation. We have identified a collection of mutants that exhibited a defective transcriptional response to nutrient limitation and failed to enter into a normal stationary phase. These rye mutants were isolated on the basis of defects in the regulation of YGP1 expression. In wild-type cells, YGP1 levels increased during the growth arrest caused by nutrient deprivation or inactivation of the Ras signaling pathway. In contrast, the levels of YGP1 and related genes were significantly elevated in the rye mutants during log phase growth. The rye defects were not specific to this YGP1 response as these mutants also exhibited multiple defects in stationary phase properties, including an inability to survive periods of prolonged starvation. These data indicated that the RYE genes might encode important regulators of yeast cell growth. Interestingly, three of the RYE genes encoded the Ssn/Srb proteins, Srb9p, Srb10p, and Srb11p, which are associated with the RNA polymerase II holoenzyme. Thus, the RNA polymerase II holoenzyme may be a target of the signaling pathways responsible for coordinating yeast cell growth with nutrient availability.

Cell Division↗

Reduction of intestinal viscosity through manipulation of dietary rye and pentosanase concentration is effected through changes in the carbohydrate composition of the intestinal aqueous phase and results in improved growth rate and food conversion efficiency of broiler chicks.

The effect of dietary rye (0, 200, 400 and 600 g/kg substituting for wheat) and pentosanase concentration (0, 1, 2, 4, 8, 16 g/kg) on weight gain, molecular weight distribution of soluble carbohydrates in the intestinal lumen and lumenal viscosity in broiler chicks was investigated. A 4 x 6 factorial design was used with four replicates per treatment and six birds per replicate pen. Diets were fed from 1 to 19 d of age, at which time body weight, food intake and intestinal viscosity and molecular weight distribution of carbohydrate complexes in proximal and distal gut sections were determined. Weight gain and food conversion efficiency (FCE) improved with increasing pentosanase and decreasing rye concentration. Intestinal viscosity, which rose as digesta passed from the proximal to distal small intestine, fell with pentosanase addition and decreasing rye concentration. Intestinal viscosity, which correlated positively with reduced weight gain and FCE, was in turn correlated with the lumenal concentration of soluble high-molecular-weight carbohydrates (HMC, greater than 500 kDa), which constituted less than 15% of the total lumenal carbohydrate concentration. The arabinose and xylose content of the HMC increased with increasing rye concentration, suggesting that HMC composition in addition to concentration may determine intestinal viscosity. The results indicate that pentosanase isolated from rye by extraction methods may not be representative of those released by digestion.

Animal Feed↗

Rye bread enhances the production and plasma concentration of butyrate but not the plasma concentrations of glucose and insulin in pigs.

The present investigation was undertaken to study the gastrointestinal and physiologic properties of diets based on soft and crisp wheat and rye breads similar in dietary fiber (DF; 230-235 g/kg dry matter) but with different proportions of the main DF polymers: in wheat, cellulose, and in rye, arabinoxylans (AX). The 2 diets provided all macronutrients; consequently, they had lower fat and sugar contents and a higher DF content than human mixed diets. The nutritional properties were studied in experiments in which pigs with cannulated ilea and catheterized portal veins and mesenteric arteries served as models for humans. The characteristics studied were degradation of nutrients, flow at the ileum, fecal output, absorption of nutrients deriving from the assimilation of cereal carbohydrates, and the insulin response. Apparent viscosity at the terminal small intestine, the ileal flow of water, flow and digestibility of noncarbohydrate constituents, but not of carbohydrates at the terminal ileum or the plasma concentrations of glucose and insulin, were higher when pigs consumed the rye compared with the wheat diet. The 2 diets provided approximately equal amounts of carbohydrates available for fermentation in the large intestine but because AX from the rye diet was more degradable than cellulose from the wheat diet, the quantitative degradation in the large intestine was more than twice as high when pigs consumed the former compared with the latter diet. The consequences included moister feces and significantly enhanced gut production and plasma concentrations of butyrate when pigs consumed the rye diet compared with the wheat diet.

Animal Feed↗

Influence of enzymes on performance and digestive parameters of broilers fed rye-based diets.

An experiment was conducted to study the influence of enzyme supplementation (ES) to rye-based diets on rate of food passage through the digestive tract, viscosity of jejunum content, volatile fatty acid concentration in ceca, and performance of broilers. There were seven treatments; six diets arranged factorially with three varieties of rye (Petkus, Prima, and Saratov V) and two levels of ES (0 or 500 ppm of an enzyme complex containing 858 IU of beta-glucanase and 864 IU of xylanase/g) and an additional control diet based on corn. Each treatment was replicated seven times (12 chicks caged together), and the trial lasted 25 d. Rye feeding increased intestinal viscosity and impaired bird performance at 25 d (P < 0.001). Among rye diets the greatest feed intake and weight gain were obtained with Petkus variety, which also produced the lowest intestinal viscosity. Enzyme addition reduced the time needed to recover 1% (0.78 vs. 0.98 h; P < 0.05) and 50% (4.2 vs. 6.5 h; P < 0.01) of the marker in feces and reduced the mean retention time of marker in the gastrointestinal tract (17.1 vs. 18.8 h; P < 0.05). Also, ES reduced intestinal viscosity (P < 0.001) and improved feed intake, daily gain, and feed conversion of birds from 4 to 25 d (P < 0.01) but did not modify volatile fatty acid concentration in ceca. We concluded that ES added to rye diets decreased intestinal viscosity and accelerated digestive transit, improving productive performance of broilers.

Animal Nutritional Physiological Phenomena↗

Randomised controlled short-term intervention pilot study on rye bran bread in prostate cancer.

The short-term effects of rye bran bread intake in prostate cancer were investigated. Ten men with conservatively treated prostate cancer were randomised to a daily supplement of 295 g of rye bran bread and eight men to 275 g of wheat bread (control) with similar fibre content for three weeks. Blood samples, ultrasound-guided core biopsies of the prostate, and urine samples were taken. In the rye group, there was a significant increase in plasma enterolactone, and the apoptotic index increased significantly from 2.1% (SD 1.3) to 5.9% (SD 1.8), P<0.005 as measured by a TUNEL index in four cases in the rye group and seven cases in the control group. Besides a significant decrease in weight in both groups, only small changes were observed in plasma concentrations of prostate specific antigen (PSA), circulating sex hormones, excreted oestrogens, insulin-like growth factor (IGF)-I, and in the endothelial fibrinolytical system. High intake of rye bran bread is suggested to increase apoptosis in prostate tumours.

Aged↗

Characterization and Quantification of Intrinsic Ice Nucleators in Winter Rye (Secale cereale) Leaves.

Extracellular ice formation in frost-tolerant organisms is often initiated at specific sites by ice nucleators. In this study, we examined ice nucleation activity (INA) in the frost-tolerant plant winter rye (Secale cereale). Plants were grown at 20[deg]C, at 5[deg]C with a long day, and at 5[deg]C with a short day (5[deg]C-SD). The threshold temperature for INA was -5 to -12[deg]C in winter rye leaves from all three growth treatments. Epiphytic ice nucleation-active bacteria could not account for INA observed in the leaves. Therefore, the INA must have been produced endogenously. Intrinsic rye ice nucleators were quantified and characterized using single mesophyll cell suspensions obtained by pectolytic degradation of the leaves. The most active ice nucleators in mesophyll cell suspensions exhibited a threshold ice nucleation temperature of -7[deg]C and occurred infrequently at the rate of one nucleator per 105 cells. Rye cells were treated with chemicals and enzymes to characterize the ice nucleators, which proved to be complexes of proteins, carbohydrates, and phospholipids, in which both disulfide bonds and free sulfhydryl groups were important for activity. Carbohydrates and phospholipids were important components of ice nucleators derived from 20[deg]C leaves, whereas the protein component was more important in 5[deg]C-SD leaves. This difference in composition or structure of the ice nucleators, combined with a tendency for more frequent INA, suggests that more ice nucleators are produced in 5[deg]C-SD leaves. These additional ice nucleators may be a component of the mechanism for freezing tolerance observed in winter rye.

Journal Article↗

Antifreeze proteins in winter rye leaves form oligomeric complexes

Antifreeze proteins (AFPs) similar to three pathogenesis-related proteins, a glucanase-like protein (GLP), a chitinase-like protein (CLP), and a thaumatin-like protein (TLP), accumulate during cold acclimation in winter rye (Secale cereale) leaves, where they are thought to modify the growth of intercellular ice during freezing. The objective of this study was to characterize the rye AFPs in their native forms, and our results show that these proteins form oligomeric complexes in vivo. Nine proteins were separated by native-polyacrylamide gel electrophoresis from apoplastic extracts of cold-acclimated winter rye leaves. Seven of these proteins exhibited multiple polypeptides when denatured and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After isolation of the individual proteins, six were shown by immunoblotting to contain various combinations of GLP, CLP, and TLP in addition to other unidentified proteins. Antisera produced against individual cold-induced winter rye GLP, CLP, and TLP all dramatically inhibited glucanase activity in apoplastic extracts from cold-acclimated winter rye leaves, and each antiserum precipitated all three proteins. These results indicate that each of the polypeptides may be exposed on the surface of the protein complexes. By forming oligomeric complexes, AFPs may form larger surfaces to interact with ice, or they may simply increase the mass of the protein bound to ice. In either case, the complexes of AFPs may inhibit ice growth and recrystallization more effectively than the individual polypeptides.

Journal Article↗

Composition of guttation fluid from rye, wheat, and barley seedlings.

This work was undertaken to determine the kinds and amount of substances that would account for the previously demonstrated differential growth of Claviceps purpurea on guttation fluids from Rosen rye, Genesee wheat, and Traill barley seedlings. Chromatographic methods were used for determining amino acids and sugars, spot tests and spectrometric methods for inorganic materials, and microbiological methods for vitamins.Total sugar content is about equal in rye and barley fluids, but lower in wheat. Glucose is the principal sugar component of the rye and barley fluids and galactose highest in wheat. Most of the amino acid in all 3 fluids is aspartic acid or asparagine. Barley fluid is far higher than the other 2 in total amino acids, with wheat the lowest. Most inorganic elements are found to be highest in barley and lowest in wheat, with the exception of iron where rye is highest and barley lowest. Barley fluid is highest in choline, p-aminobenzoic acid, thiamine, and uracil, while rye is highest in inositol and pyridoxine. Wheat is much lower than the other 2 in choline and inositol.

Journal Article↗

Cyclic adenosine 3':5'-monophosphate in axenic rye grass endosperm cell cultures.

Cyclic adenosine 3':5'-monophosphate (cAMP) was extensively purified from rye grass (Lolium multiflorum) endosperm cells grown in axenic suspension culture. The cAMP was purified by neutral alumina and anion and cation exchange chromatography. The cAMP was quantitated by means of a radiochemical saturation assay using a beef heart cAMP-binding protein and also by an assay involving activation of beef heart protein kinase. The cAMP levels found (corrected for recovery of tracer cyclic 3',5'-[8-(3)H]AMP included from the point of sample extraction) ranged from 2 to 12 pmol/g fresh weight. The material purified from rye grass cultures was indistinguishable from authentic cAMP with respect to chromatography in two cellulose thin layer systems, behavior on dilution in both the saturation and protein kinase activation assays, and rates of degradation by a mammalian cAMP phosphodiesterase. The cAMP from rye grass cultures was completely degraded by a mammalian cAMP phosphodiesterase, and 1-methyl-3-isobutylxanthine inhibited such degradation. The protein kinase activation and saturation assays gave essentially the same values for the cAMP content of axenic rye grass culture extracts. Material satisfying the above criteria for identity with cAMP was also isolated from the culture medium. The increase observed in medium cAMP levels during culture growth provides evidence for the synthesis and secretion of cAMP by rye grass endosperm cells in suspension culture.

Journal Article↗

Barley and rye prolamins induce an mRNA interferon-gamma response in coeliac mucosa.

BACKGROUND: In coeliac disease, wheat, barley and rye are traditionally excluded in the gluten-free diet. However, few studies have examined the small intestinal immune response to barley and rye. AIM: To investigate the immunogenicity of barley and rye prolamins (hordein and secalin respectively) in comparison with wheat gliadin. METHODS: Duodenal biopsies from 22 coeliac patients and 23 disease controls were cultured for 4 h with gliadin, hordein or secalin and compared with culture medium alone. Proinflammatory cytokines, interferon-gamma and interleukin-2, were quantified by TaqMan polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: Hordein caused the greatest increase in interferon-gamma mRNA in coeliac patients (median: 3.3-fold) in comparison with control subjects (median: 0.28-fold, P < 0.085). Secalin and gliadin induced similar levels of interferon-gamma mRNA with median fold-changes of 3.4 and 2.8, respectively, in coeliac patients in comparison with 1.6- and 1.1-fold increases in control subjects (P < 0.294 and P < 0.105, respectively). The median fold-changes for interleukin-2 mRNA did not differ between coeliac patients and controls. Cytokine protein was not upregulated. CONCLUSION: The findings of this study provide evidence that barley and rye cause immune activation in the mucosa of coeliac patients and support the practice that barley and rye should be excluded from the gluten-free diet.

Adult↗

Immunoglobulin E specific to wheat and rye flour proteins.

We have used the radioallergosorbent test (RAST) to determine IgE antibodies specific to wheat flour proteins in the sera of seven groups of patients. In some cases rye-specific IgE was also determined. Wheat and rye RAST scores showed a good correlation, presumably due to cross-reactions. Among bakers with asthma, positive scores, 0.5-3, occurred with a prevalence of 43%, and among children with eczema, scores in the range 0.5-4 were found with a prevalence of 54%. A score of 0.5 was a marginal value which was also occasionally encountered with sera from patient groups with no history of immediate hypersensitivity to wheat or rye. These groups included adults and children with allergic rhinitis and asthma, children from the general population and children with coeliac disease. The RAST appeared useful in the diagnosis of allergy to inhaled flour dust among bakers. Among children with eczema, positive wheat and rye RAST results were a common finding, which only occasionally could be linked to strong and unequivocal reactions to the foods in question. Both in bakers and children with eczema, wheat and rye RAST results showed good agreement with intracutaneous skin test results.

Adult↗

Activation of the alternative pathway of complement by grain. I. C3PA conversion and quantification of complement consumption by rye.

Ground whole rye and airborne rye dust of comparable size distribution were tested for their ability to activate the complement cascade via the alternative pathway. Precipitin-negative pooled normal human serum was incubated with increasing amounts of the two rye dusts. Electrophoresis of the resultant supernatant fluids demonstrated the conversion of the proactivator of the third component of complement to the gamma-migrating activator of the third component. This activation was completely prevented by pre-treating the serum with the chelator EDTA, while pre-treatment with EGTA allowed suboptimal arc conversion, strongly implying that complement was activated via the alternative pathway. Quantification of the supernanant fluids showed dose-dependent complement consumption as defined by both CH100 immunodiffusion and CH50 tube haemolytic techniques. Airborne rye dust showed a greater quantitative potential than ground whole rye for activating the alternative pathway. These results indicate the possibility of the direct action of airborne organic dusts on the induction of inflammatory sequelae in the lungs of both sensitized and unsensitized individuals.

Air Pollutants↗

Identification of crossreacting wheat, rye, barley and soya flour allergens using sera from individuals with wheat-induced asthma.

We have conducted radio allergosorbent test (RAST), competitive RAST inhibition, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting using sera from patients with wheat-induced asthma to investigate the immunological relationship between wheat, rye, barley and soya, and to identify common proteins between these flours. RAST showed strong associations between the levels of specific IgE to wheat flour and those of rye and barley flour. Competitive RAST inhibition showed that wheat, rye, barley and soya flours contained crossreacting proteins, in decreasing concentrations. Wheat, rye and barley flours had similar protein profiles on gel electrophoresis. Soya flour contained a number of high molecular weight proteins not present in the other cereals. Western blotting using sera from 21 wheat flour hypersensitive individuals identified a large number of allergens in the different flours. Proteins of 69, 33, 26, 21 and 12 kDa were identified as major wheat flour allergens. Rye flour proteins of 21 and 12 kDa, and barley flour proteins of 69, 52 and 10 kDa were the major allergens identified by serum from wheat hypersensitive individuals. The major common protein of soya and wheat flour had a molecular weight of 21 kDa. The majority of crossreacting allergens identified between the different flours have molecular weights similar to those of known flour enzymes or enzyme inhibitors.

Allergens↗

Associated mycoflora of rye bread.

Penicillium roqueforti (27%), P. corylophilum (20%) and Eurotium species (15%) made up the important mycoflora associated with rye bread on 3425 identified fungi isolates. These fungi were dominant as spoilers of packaged rye bread in almost every month of a 4 year investigation. Penicillium decumbens (3%), Paecilomyces variotti (8%) and Aspergillus flavus (5%) were found more rarely, but were the major species found over periods of a few months. Penicillium commune (5%), P. solitum (4%), A. niger (4%) and Mucor species (2%) were a constant, but small, part of the total mycoflora of rye bread. Identification of the fungi in the production environment in a rye bread factory showed the locality of potential contamination sources. Elimination of the contamination sources by improved cleaning and disinfection procedures quickly resulted in a significant reduction in the frequency of mould growth in the packaged rye bread.

Aspergillus flavus↗