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PubMed1986

Monoclonal antibodies to the major Lolium perenne (rye grass) pollen allergen Lol p I (Rye I).

Thirteen monoclonal antibodies (MAbs) were produced against Lol p I (Rye I), the major Lolium perenne (rye grass) pollen allergen. Spleen cells from A/J and SJL mice immunized with highly purified Lol p I (Lol I) were allowed to fuse with cells from the non-secreting Sp2/0-Ag14 myeloma cell line. Each MAb was analyzed for antigenic specificity by radioimmunoassay (RIA) using 125I-Lol I. The epitope specificities of seven of the MAbs were examined by competitive binding against a labelled standard MAb for the Lol I antigen (Ag). The dissociation constant, Kd, of one MAb (No. 3.2) that was studied most extensively was determined by double Ab RIA to be 3.5 X 10(-6) L/M. This MAb recognized the related 27,000-30,000 Group I glycoproteins found in the pollens of nine other species of grass pollens tested, including weak binding to Bermuda grass Group I (Cyn d I), which by conventional analysis using polyclonal anti-Lol I serum shows no detectable binding. Monoclonal antibody No. 3.2 was coupled covalently to Sepharose 4B and used to prepare highly purified Lol I from a partially purified rye pollen extract. Finally, an RIA was developed which permitted the analysis of the Group I components in rye grass and nine other grass pollen species. The latter assay is likely to prove useful in the standardization of grass pollen extracts according to their Group I contents.

Allergens↗
PubMed2005

Cell wall fractions isolated from outer layers of rye grain by sequential treatment with alpha-amylase and proteinase: structural investigation of polymers in two ryes with contrasting breadmaking quality.

Recent studies have indicated that some structural features of arabinoxylans, the major cell wall polysaccharides, might be potential quality markers in the selection of rye breeding materials. To specify the most appropriate characteristics, the differences in the structure of cell wall components were studied in two ryes with high and low breadmaking qualities. Two cell wall fractions were isolated from the outer layers of the grain (pooled shorts and bran fractions) by a consecutive water extraction with alpha-amylase (WE-A) and proteinase K (WE-P). Polysaccharides predominated in the WE-A fraction (approximately 64%, mainly arabinoxylans). By contrast, the WE-P fraction contained mostly protein (approximately 63%), and its level of polysaccharides was relatively low (approximately 18%). The 1H NMR and sugar analysis of the ammonium sulfate precipitated subfractions revealed that the WE-A was built of four arabinoxylan populations with marked structural differences (arabinose-to-xylose ratios, Ara/Xyl, of approximately 0.3, 0.5, 0.8, and 1.2). Instead, the arabinoxylans present in the WE-P were generally enriched in disubstituted xylopyranosyl residues. The ratio of phenolic components to arabinose residues in the WE-P fraction (indicated by 1H NMR) and the proportion of polymers with the highest molecular weights in the WE-A fraction (revealed by HPSEC) distinguished well two ryes with diverse breadmaking qualities. Much less obvious differences between both ryes were observed in the ratio of amide I to amide II band intensities of FTIR spectra for the WE-P and in the level of phenolic acids and ferulic acid dehydrodimers for both cell wall preparations.

Arabinose↗
PubMed1986

[Phytic acid and cereals and cereal products. I: Phytic acid and phytase in rye and rye products].

Phytic acid in food is considered to be responsible for a reduced bioavailability of essential dietary minerals; its detrimental effects can be diminished by hydrolysis with phytase during processing. The average phytic acid content was 8.18 mg/g and 3.44 mg/g and average phytase activity was 3.7 U/g and 2.6 U/g in rye kernels and in flour (Type 997, 1.09 ash content), respectively. Phytate and Phytase were about equally distributed between the two kernel halves (cross sections). During the early stages of germination (3 days) phytase activity did not change, and phytic acid content was reduced to 67%. After milling most of the phytic acid and phytase activity were found in the bran fractions. It is concluded that substrate and enzyme are present in the same kernel structures but separate within the cells. Cooking of ground rye caused a phytate hydrolysis which was the more effective 1.) the smaller the particle sizes were, 2.) the more water was added, and 3.) the longer phytase worked at optimum temperature. Extrusion cooking of the rye whole flour at up to 100 degrees C did not influence the phytic acid level but caused a 23% reduction at 170 degrees C. Phytase activity was reduced by 80% by extrusion cooking at 80 degrees C.

6-Phytase↗
PubMed2003

Evaluation of rye (Secale cereale L.) inbred lines and their crosses for tissue culture response and stable genetic transformation of homozygous rye inbred line L22 by biolistic gene transfer.

The efficient and reproducible production of stably expressing transgenic rye plants is described. Analysis of the genotype-specific callus culture-response of 21 rye inbred lines, single crosses and a population variety resulted in the identification of the highly responsive inbred line L22. Biolistic transformation experiments were performed using line L22 and the impact of different selection agents on the regeneration capacity was analyzed. Using the selectable marker gene nptII and corresponding paromomycin selection resulted in transformation efficiencies of up to 4.0% of the bombarded explants. A total of 17 independent transgenic rye plants were produced, and their stability and level of transgene expression was analyzed. The majority of these lines showed stable transgene expression. In contrast a few transgenic lines with multiple transgene inserts, provided evidence of transcriptional and post-transcriptional gene silencing.

Biolistics↗
PubMed2002

Quantitation of odor-active compounds in rye flour and rye sourdough using stable isotope dilution assays.

Application of the aroma extract dilution analysis on a flavor distillate prepared from freshly ground rye flour (type 1150) revealed 1-octen-3-one (mushroom-like), methional (cooked potato), and (E)-2-nonenal (fatty, green) with the highest flavor dilution (FD) factors among the 26 odor-active volatiles identified. Quantitative measurements performed by stable isotope dilution assays and a comparison to the odor thresholds of selected odorants in starch suggested methional, (E)-2-nonenal, and hexanal as contributors to the flour aroma, because their concentrations exceeded their odor thresholds by factors >100. Application of the same approach on a rye sourdough prepared from the same batch of flour revealed 3-methylbutanal, vanillin, 3-methylbutanoic acid, methional, (E,E)-2,4-decadienal, 2,3-butanedione, and acetic acid as important odorants; their concentrations exceeded their odor thresholds in water and starch by factors >100. A comparison of the concentrations of 20 odorants in rye flour and the sourdough made therefrom indicated that flour, besides the fermentation process, is an important source of aroma compounds in dough. However, 3-methylbutanol, acetic acid, and 2,3-butanedione were much increased during fermentation, whereas (E,E)-2,4-decadienal and 2-methylbutanal were decreased. Similar results were obtained for five different flours and sourdoughs, respectively, although the amounts of some odorants in the flour and the sourdough differed significantly within batches.

Aldehydes↗
PubMed2002

Enzymatic solubilization of arabinoxylans from isolated rye pentosans and rye flour by different endo-xylanases and other hydrolyzing enzymes. Effect of a fungal caccase on the flour extracts oxidative gelation.

Water-extractable (WEP) and water-unextractable (WUP) pentosans were isolated from a rye flour. The effect of a commercial enzyme preparation, Grindamyl S 100 (GS100), containing pentosanase activities, was investigated on WEP, WUP, a mix of WEP and WUP, and the rye flour, with the aim to monitor the solubilization and depolymerization of high molecular weight arabinoxylans and the effect on the viscosity of the reaction medium. The effects of other hydrolyzing enzymes were also tested. Three xylanases were used: xylanase 1 (Xyl-1) from Aspergillus niger, the main activity present in GS100; xylanase 2 (Xyl-2) from Talaromyces emersonii; and xylanase 3 (Xyl-3) from Bacillus subtilis. Xyl-3 was used in combination with Xyl-1, (1,4)-beta-D-arabinoxylan arabinofuranohydrolase, endo-beta-D-glucanase, or ferulate esterase from A. niger, but no synergism was observed. GS100 and xylanases increased the arabinoxylan solubilization, Xyl-3 and Xyl-1 being those that presented the best yields of extraction without extensive depolymerization of water-extractable arabinoxylans. Both xylanases were affected by an inhibitor in rye flour. Flour treated with hot ethanol was used to study the oxidative gelation of flour extracts treated with xylanases, in the presence of laccase from Pycnoporus cinnabarinus. Two doses of xylanases were tested (0.5 and 2.5 units). Only the flour extracts treated with 0.5 unit of Xyl-1 thickened.

Carboxylic Ester Hydrolases↗
PubMed1999

[Efficacy of non-starch polysaccharide hydrolyzing enzymes on nutrient digestibility and gross energy convertibility of barley-rye and wheat-rye based diets for laying hens].

The efficiency of beta-glucanase or xylanase containing enzymes supplemented to rations for laying hens on the basis of barley-rye and wheat-rye were tested in two balance experiments. Five diets for eight hens each (White Lohmann Leghorn Hybrids) have been proved. The balances were carried out in the 23rd and 29th week of life. Statistically significant differences in the digestibility of nutrients like protein, fat and starch as well in the transformation of gross energy and metabolizable energy appeared especially in the first balance period. In the second balance period the starch digestibility was increased significantly from 93.7% to 99.7% by enzyme supplementation of the barley-rye rations. A positive influence of enzymes on the ileal digestibility of starch and fat could also be found. The addition of enzymes led to a decreasing reaction of the viscosity in the feed, in the different parts of the digesta and in the excreta.

Animal Feed↗
PubMed1992

Allergens from rye pollen (Secale cereale). I. Study of protein release by rye pollen during a 19-hour extraction process. Allergen identification.

We have studied the proteins and allergens released by rye pollen in the course of a 19-h pollen incubation process. Nearly 40% of the total extracted proteins were collected during the first 5 min, and most of them had a molecular weight less than 28 kDa. Between 5 and 30 min, 15% of the proteins from total extract were released, showing in the SDS-PAGE analysis an increase in which components moved close to 30 kDa standard. From 30 min to 19 h several extracts were collected. Electrophoretical profile of components from these extracts reveals that bands moving below 28 kDa were practically absent and those of 28 and 23 kDa became very intense. At the end of the process there was a rise of 67 kDa proteins. Dot-immunobinding and immunoblotting techniques reveal that allergens leave the rye pollen, for the most part, after 5 min incubation and are proteins with 28 kDa, 33 kDa, 48 kDa and 67 kDa molecular weights.

Allergens↗
PubMedDate not supplied

[Cytological markers of rRNA gene expression during microsporogenesis in rye, wheat and wheat-rye hybrids].

The quantity of nucleolei during microsporogenesis of wheat, rye and F1 of wheat-rye hybrids has been investigated. Dependence of nucleoli quantity from microsporogenesis stage and number of nucleolar organizer regions in chromosomes have been shown. The volumes of nuclei and nucleoli as well as nucleus-nucleolus ratio have been calculated. The changes of these indices during microsporogenesis were regular. The possibility of using cariometrical indices as cytological markers of rRNA gene expression in the process of microsporogenesis has been substantiated.

Cell Nucleolus↗
PubMed1982

Effects of processed rye bran and raw rye bran on glucose metabolism in alloxan diabetic rats.

Alloxan diabetic rats (D) and control rats (C) were for 14 days fed a basic diet made from a mixture of rye bread and milk containing either no bran (group O), processed bran (group PB) or raw bran (group B). The D-B animals has a lower urinary glucose excretion (36 +/- 4 mmoles/24 hours) than the D-PB animals (63 +/- 5 mmoles/24 hours), which in their turn had a lower excretion than the D-0 animals (100 +/- 3 mmoles/24 hours). The effects of the two brans on weight, serum insulin levels, and non-fasting blood glucose levels were similar in D-rats as well as in C-rats, with the exception of the lower nonfasting blood glucose level in C-B animals than in C-PB animals.

Animals↗
PubMed2001

[Effect of rye chromosomes on features of androgenesis in wheat-rye substituted lines of Triticum aestivum L. sort Saratovskaya 29/Secale cerale L. sort Onokhoiskaia and Triticale].

The characteristic features of androgenesis in six wheat-rye substitution lines Triticum aestivum L. (cv. Saratovskaya 29)/Secale cereale L. (cv. Onokhoiskaya) and triticale (2n = 56) using anther culture at different concentrations of 2,4-D in the growth medium were studied. Under variable cultivation conditions, the significant effect of genotypic diversity on the variability of such androgenesis parameters as the frequency of productive anthers, the frequency of embryoid formation, and the frequency of total regenerated plantlets, was shown. It was demonstrated that chromosomes 1R, 3R, and 7R stimulated the formation of androgenous embryoids, while chromosome 5R produced an opposite effect. In triticale and substitution lines, the regeneration ability of androgenous embryoids induced by elevated 2,4-D concentrations was inhibited. Chromosome 1R of the Onokhoiskaya cultivar was suggested to contain genes suppressing regeneration of green plantlets, while chromosome 3R, conversely, stimulated their formation. Chromosomes 1R, 2R, 3R, and 7R of the Onokhoiskaya cultivar did not inhibit the spontaneous formation of androgenous hexaploids in the substitution lines.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1991

[Photosystem II of rye. Nucleotide sequence of the psbB, psbC, psbE, psbF, psbH genes of rye and chloroplast DNA regions adjacent to them].

In order to determine structures of the barley photosystem II subunits, the following genes have been cloned: psbB, encoding 47 kDa chlorophyll-binding subunit; psbH, encoding 7.7 kDa phosphoprotein; psbE and psbF, encoding 9.3 and 4.4 kDa subunits of the cytochrome b559 apoprotein, respectively; and a fragment of psbC gene, encoding the 43 kDa chlorophyll-binding subunit. The nucleotide sequences of these genes and the deduced amino acid sequences of their products are highly homologous to the corresponding sequences for other plant species.

Chlorophyll↗
PubMed1983

Influence of rye and dietary salt level on water and sodium metabolism in intact and colostomized roosters.

A study of water and Na metabolism was conducted with 24 roosters fed diets based on either corn or rye with two levels (.4 and 1.0%) of added salt. A diet containing water-extracted rye and a diet based on corn with a freeze-dried water extract of rye added at a level of 12% were also used in the study. Measurements were made on intact and colostomized roosters. In intact roosters dry matter metabolizability was depressed when rye diets were compared with corn diets. Measurements in feces from colostromized roosters fed rye as compared to those fed corn indicated that moisture, viscosity, and water holding capacity were increased and that bulk density was decreased. Diet salt level did not influence the responses regarding these parameters, but water extraction of rye completely alleviated the effects of rye on dry matter metabolizability and viscosity and partially reduced the influence of rye on fecal moisture while not altering the responses regarding water holding capacity or bulk density of feces. With the exception of feces moisture, the water extract of rye had no influence regarding these parameters. Roosters fed rye in comparison to those fed corn showed increases in water intake and excreta moisture content and decreases in urine sodium concentrations and plasma clearance of sodium. The response to rye regarding these latter parameters was dependent on diet salt level. The data indicated a reduced availability of sodium at the cellular level in rye-fed birds, which was manifest in a difference in route of excretion of sodium such that urine was the major route in corn-fed birds and feces was the major route for birds fed rye. The dietary fiber content of rye appeared to be a major factor in rye influencing sodium availability.

Animals↗