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Construction of a chloroplast DNA library from rye (Secale cereale), a cereal plant of temperature-inducible chloroplast ribosome deficiency.

The chloroplast genomes of flowering plants are circular DNA molecules, 120 to 160 kilobase pairs long, encoding the rRNA, all tRNAs, and 21 r-proteins of the chloroplast translational apparatus as well as key protein components of the photosynthetic and carbon reduction cycle reactions. In this paper we describe some characteristics of the rye chloroplast (plastid) genome and the construction and characterization of a clone library of 93% of its DNA in a plasmid and a cosmid vector. The size of rye chloroplast DNA is estimated at 135 kbp, similar to that for wheat and rice but slightly smaller than the estimate for maize (139 kbp). Chloroplast ribosome deficiency is induced in rye seedlings by germination and growth at 32 degrees-34 degrees C; therefore these clones would be useful for analyzing the regulation of chloroplast ribosome synthesis in higher plants, a process that requires coordinate expression of genes located in the nucleus and the chloroplast.

Chloroplasts↗

[Effect of plant residues on the parasitic activity of soilborne pathogens and the saprophytic microflora of the soil. III. Influence of rye and rape grown as winter catch crop on the incidence of Cercosporella herpotrichoides Fron (author's transl)].

In a five-years field trial, the influence of two winter catchcrops, rye and rape, upon the incidence of Cercosporella herpotrichoides has been studied. The winter catch-crops had been repeated three times, followed either by stubble-crop or without in fall of the fourth year. The variants (blocks) had been completed by two variants without catch-crop, either carefully cultivated or overgrown with weeds. Each of the four blocks had been divided three times to be grown with three different crop sequences containing 80%, 60% (with the insusceptible corn), or 40% haulm fruit, respectively. Each of the 12 variants had been divided into two parts by continuous fallow stripe, one part being artificially infested with the pathogen in the beginning (1st winter wheat). In the 2nd, 3rd, and 4th year, only spring crops were grown, followed by winter wheat in the 5th year. The final exploration resulted in a medium to heavy degree of infection, being significantly higher in the 80% cereal crop sequence than in the 60% and 40% sequences. Concerning the catch-crops, infection in the rye variants was equal or significantly lower than in the carefully cultivated variants without catch-crop. On the other hand, infection of the rape variants had become significantly higher than that of the catch-crop-free ones, about as heavily as in the weed-infested variants. From fall 1966 to summer 1969, the development of the saprophytic soil microflora and several other edaphic parameters had been investigated. Remarkable differences in intensity of disintegration have been observed under the influence of the main crops, cereals or potatoes, respectively, but no differences could be stated resulting from the matter of disintegration--residues of rye or rape--, which might interpret the contrasting effects on the pathogen. Too, no indication for antagonistic activity of bacteria or actinomycetes has been noted. The results are compared with known facts of Cercosporella and of other soil borne pathogens. The difference in biology of Cercosporella on the one hand, and of root infecting fungi on the other hand seems to be most important to explain the results obtained.

Actinomycetales↗

[Immunochemical characterization of proteolytic enzymes (BAPAase) from buckwheat and rye seeds].

A comparative study of the immunochemical properties of two serine proteolytic enzymes (BAPases) from buckwheat and rye seeds hydrolyzing N alpha-benzoyl-DL-arginine-p-nitroanilide (BAPA) has been performed. It has been shown that buckwheat and rye seed BAPAases were partially antigenically identical. This was demonstrated using double immunodiffusion in agar and inhibition of one enzyme with an antiserum to the other. At the same time neither of the antisera inhibited trypsin. Thus, buckwheat and rye seed BAPAases have common antigenic determinants and, consequently, common structural features. On the other hand, these enzymes probably have no common structural elements with trypsin.

Benzoylarginine Nitroanilide↗

Studies on lectins. LII. Isolation and characterization of the lectin from rye germ (Secale cereale L.).

Rye germ lectin was isolated by extraction of defatted rye germ, fractionation of the extract by ammonium sulfate precipitation, affinity chromatography of the active substances on chitin-beta-glucan and gel filtration on Sephadex G-50. The lectin shows erythroagglutinating activity at a minimum concentration of 2.5 micrograms/ml. The erythroagglutinating activity is the same against human red blood cells of all types of the ABO system and is inhibited by N-acetyl-D-glucosamine. The lectin has no mitogenic activity against mouse splenic lymphocytes. According to the results of polyacrylamide gel electrophoresis the lectin obtained is a mixture of three very similar isolectins of equal erythroagglutinating activity. Sedimentation analysis indicates homogeneity of the lectin preparation; the molecular weight was 56000 as estimated by sedimentation equilibrium. In the presence of urea and sodium dodecyl sulfate the lectin dissociates into 2 types of subunits with molecular weights of 35000 and 19000. The rye germ lectin contains about 2% of neutral sugar and 1% of D-glucosamine. The amino acid composition of the lectin is characterized by a very high content of glycine and half cystine and a low content of apolar amino acids. N-terminal amino acids of the lectin are apparently blocked.

Acetylglucosamine↗

[Difficulties in the application of Rye's classification to Hodgkin lymph nodes].

Rye's histological classification has been used the world over for the last 15 or so years in Hodgkin's disease but its practical application still poses numerous problems. These are related either to the border-line cases which separate the 4 Rye types or to the sub-classification of nodular sclerosis, or the problem of deciding whether certain classical or recently described histological entities can indeed be called Hodgkin's disease. The Rye classification allows a better approach to a positive and differential diagnosis of Hodgkin's disease and its prognostic and epidemiological usefulness is indisputable. This is to say that the criteria which define the four types should be more specific leaving less for subjectivity. Meanwhile, a collegial analysis of the histological data definitely improves the reliability of this system.

Hodgkin Disease↗

Excretion of amino acid residues from diets based on low-fibre wheat or high-fibre rye bread in human subjects with ileostomies.

OBJECTIVES: The effect of consumption of a wheat-flour bread-based low-fibre diet (13.6 g/day fibre per portion) or a rye bran bread and whole-grain rye crispbread-based high-fibre diet (43.5 g/day fibre per portion) on the ileal excretion of amino acid residues and crude protein was studied. DESIGN: The study was performed as a cross-over design. SETTING: The subjects were studied as outpatients except on the sampling days when all subjects were admitted to the research ward and stayed in a nearby patient hotel overnight. SUBJECTS: Six men and two women, all proctocolectomised for ulcerative colitis, volunteered to participate in the study. INTERVENTIONS: During each dietary period of 3 weeks, food and excreta were collected and analysed on days 3, 17 and 18. RESULTS: The excretion of crude protein and amino acids (free and bound) was generally higher during the high-fibre diet period, although the relative proportion of amino acids was similar in the ileal effluents from the two diets. The apparent digestibilities of crude protein and all amino acids except histidine, tyrosine and proline were significantly (P < 0.05) lower during the high-fibre diet period. CONCLUSION: It was indicated that the effect of the high-fibre rye-based diet was a generally impaired ileal amino acid absorption.

Aged↗

Intestinal degradation in pigs of rye dietary fibre with different structural characteristics.

In order to investigate the effects of dietary fibre (DF) characteristics on carbohydrate degradation and the metabolism in the large intestine, pigs were fed on four rye-bread diets (based on whole rye, pericarp/testa, aleurone or endosperm) with differences in characteristics and amount of DF. The degradability of DF in the large intestine varied greatly between diets. The pericarp/testa DF was hardly degraded in the large intestine, whereas endosperm DF was extensively and rapidly degraded in the caecum. Caecal degradation of aleurone DF was also limited, leaving more material to be degraded in the colon. The undegradable pericarp/testa DF was characterized by high contents of lignin, cellulose, ferulic acids and highly substituted arabinoxylans (the major DF component in rye). Ingestion of this diet resulted in increased faecal bulk and reduced transit time, but with low colonic pH and the lowest concentrations of shortchain fatty acids (SCFA). The aleurone diet, on the other hand, led to a fermentation pattern which may be considered more optimal, with lower colonic pH and higher concentrations of SCFA, in particular butyric acid. Despite the large differences in carbohydrate fermentation only minor significant effects on the presence of protein degradation products and on histological measurements (height and diameter of colonic crypts and thickness of the muscularis external) were observed.

Animals↗

Flow cytometric DNA analysis of wheat-rye addition lines.

The nuclear DNA content of the 7 wheat-rye chromosome addition lines were determined by flow cytometry. The nuclear DNA content of each line was compared to the original wheat parental line. From this information, the amount of DNA per rye chromosome was calculated. The DNA per chromosome ranged from 0.8 picograms (pg) in chromosome 3R to 1.4 pg in chromosome 7R. Flow cytometry was proven to be a rapid and reliable technique for the evaluation of aneuploid plant populations. Flow cytometry has the potential to allow for more rapid screening of aneuploid and euploid cytological stocks than conventional cytogenetic techniques.

Aneuploidy↗

Evaluation of sensitivity of flow cytometry in detecting aneuploidy in wheat using disomic and ditelosomic wheat-rye addition lines.

Flow cytometric DNA analysis was used to study changes in nuclear DNA content induced by the addition of complete or telosomic rye chromosomes into the genome of common wheat (Triticum aestivum L.). The DNA content of each addition line was determined by comparison with an internal reference value and was expressed as a difference with respect to the original wheat parental line. A 1.84% difference in the DNA content could be detected. Nuclei were flow sorted and the presence of rye chromatin in the nuclei with the higher DNA content was demonstrated by Southern hybridization. Flow cytometry was proven to be sensitive enough to detect the small DNA content deviations that are expected to occur in aneuploid plants of wheat.

Aneuploidy↗

A new technology for the production of valuable vitamin extracts from wheat and rye germs.

A method for nearly complete extraction of the lipid fraction from wheat and rye germs was developed, and the technological parameters of the production process were specified. The resulting preparations are extracts rich in vitamin E and other biologically active components. Dermatological studies and application in the cosmetic industry have testified to the suitability of these preparations for use in cosmetic and pharmaceutical products. Owing to their high content of unsaturated fatty acids and to a natural combination of biologically active compounds, the preparations from wheat and rye germs obtained by our method on an industrial scale find increasingly wider application. The post-extraction residue free from the lipid fraction is high-value feed.

Chemical Phenomena↗

The use of freeze-substitution and LR gold in the study of rye grass (Lolium perenne) pollen.

Pollen grains of Lolium perenne (rye grass) were prepared for transmission electron microscopy by rapid freezing in liquid propane, substitution in acetone, methanol or diethyl ether, and embedment in the acrylic resin London Resin gold. These were compared to pollen chemically fixed (CF) in aldehyde/osmium tetroxide and embedded in the epoxy resin Quetol 651. Ultrastructural preservation was superior in freeze-substituted (FS) pollen, particularly with the use of acetone or methanol. Optimally preserved FS pollen displayed a homogeneous aspect of the cytoplasm and nucleoplasm, and smooth, uninterrupted contour or organelles. A striking difference was also seen in the preservation of inclusions in the intine. Varied forms and sizes of intine inclusions were evident in FS pollen but these were not discernible in the CF image. The FS scheme studied here presents enormous potential for both ultrastructural and immunolabelling studies in rye grass pollen. Problems discussed include artifacts associated with each of the substitution solvents used, and a gradient of freezing damage observed within the pollen grain.

Acrylic Resins↗

Identification of barley and rye varieties using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry with neural networks.

Cereal varieties are normally identified using time-consuming methods such as visual examination of either the intact grain or one-dimensional electrophoretic patterns of the grain storage proteins. A fast method for identification of wheat (Triticum aestivum L.) varieties has previously been developed, which combines analysis of alcohol-soluble wheat proteins (gliadins) using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry with neural networks. Here we have applied the same method for the identification of both barley (Hordeum vulgare L.) and rye (Secale cereale L.) varieties. For barley, 95% of the mass spectra were correctly classified. This is an encouraging result, since in earlier experiments only a grouping into subsets of varieties was possible. However, the method was not useful in the classification of rye, due to the strong similarity between mass spectra of different varieties.

Glutens↗

A moderately repeated DNA sequence of wheat and rye genomes.

A 371 base pair segment (bordered by Hind III and Eco RI cutting sites) of wheat embryo nuclear DNA has been cloned and sequenced. It is AT-rich (68%), shares some sequence features with autonomously replicating sequence (ARS) elements, and occurs in approximately 7600 copies per haploid genome. When used as probe for blot hybridization to Hind III-digested wheat DNA, it gives an irregular series of hybridization bands. Essentially the same hybridization pattern was observed for rye DNA. It is concluded that this segment is distributed irregularly but, apparently, according to the same rule in both wheat and rye genomes.

Base Sequence↗

Physical mapping of 5S rDNA reveals a new locus on 3R and unexpected complexity in a rye translocation used in chromosome mapping.

Using fluorescence in situ hybridization (FISH) with probe pScT7, three different 5S rDNA loci were detected in the satellite of rye chromosome 1R (5SDna-R1) and in the short arms of chromosomes 3R (5SDna-R3) and 5R (5SDna-R2) respectively. All three loci showed polymorphism for the hybridization signal intensity. In order to determine the localization of these rye 5S rDNA multigene loci with higher precision within the corresponding chromosome arms, the probe pScT7 was physically mapped by FISH in relation to the following five translocations (Wageningen Tester Set): T850W (1RS/4RL), T248W (1RS/6RS), T273W (1RS/5RL), T305W (2RS/5RS) and T240W (3RS/5RL). Accurate physical maps of the translocation breakpoints had previously been made using electron microscope analysis of spread pachytene synaptonemal complexes of heterozygotes for the different translocations. The results indicate that locus 5SDna-R3 is located between the breakpoint of translocation T240W and the telomere, whereas locus 5SDna-R2 is located between the breakpoint of translocation T305W and the centromere, the hybridization of probe pScT7 on T305W translocated chromosomes demonstrating the complex nature of this translocation. On the other hand, the simultaneous detection of probes pScT7 and pTA71 (18S-5.8S-26S rDNA) with two different fluorochromes, indicated that the breakpoints of translocations T850W and T248W are located between loci Nor-R1 and 5SDna-R1.

Chromosome Mapping↗

The influence of B chromosomes on rDNA organization in rye interphase nuclei.

Patterns of rye rDNA organization in interphase nuclei were studied through the use of in situ hybridization in spreads of root meristem cells from plants with and without B chromosomes (Bs). In cells from plants without Bs each rDNA locus is organized as a single perinucleolar knob of condensed chromatin with decondensed chromatin inside the nucleolus. In plants with Bs there is a marked modification of the pattern, found in more than 23% of nuclei, which involves several regions of condensed chromatin interspersed with decondensed chromatin inside the nucleolus. This B-induced alteration in rDNA interphase organization suggests a change in expression of the rRNA genes located on the A chromosomes probably related to the reduction in nuclear RNA observed previously in plants with Bs. The influence of the Bs on the expression of A chromosome genes, through rearrangement of interphase chromatin, could provide the basis of an explanation for some of the known phenotypic effects of B chromosomes in rye.

Cell Nucleus↗

Immunocrossreactivity of antisera against grain prolamines of wheat, rye, barley and oats.

Prolamines extracted from wheat (gliadin), rye (secalin), barley (hordein) and oats (avenin) were used to raise antibodies in rabbits. The four prolamines were separated by SDS-PAGE and blotted onto nitrocellulose. The immunocrossreactivity of the separated prolamines with the four antisera was studied. The close botanical relationship between wheat and rye, and to a lesser extent barley, is clearly established. The crossreactivity of gliadin, secalin and hordein with anti-avenin serum was found to be weak. In contrast, avenin shows a strong crossreactivity with anti-gliadin serum.

Animals↗

Activity tests of alcohol dehydrogenases in wheat, rye and triticale.

The relative band staining intensities of ADH isoenzymes in wheat and triticale suggest alloploid genome interactions. Rye ADH is scarecely affected by anti-wheat-ADH. Despite the evolutionary divergence of their Adh genes, ADH monomers of wheat and rye form enzymatically active heterodimers in triticale.

Alcohol Oxidoreductases↗

[Protein quality of flatbread and extruded dry flatbread made from full-grain rye groats].

The effect of various processing methods used for the production of dry flatbread from ground whole rye on the nutritional protein quality were investigated by means of the nitrogen balance method in growing rats. The following products were tested: untreated ground whole rye, baked crisp bread, extruded flatbread without and with the addition of 2.5% and 5.0% of saccharose. The apparent protein digestibility (PD) of the 5 samples varied between 69% and 72% and was neither affected by the method of processing nor by the difference in sugar content. Compared to the untreated raw material (NPU = 41%; BV = 59%) net protein utilization and biological value in crisp bread baked by the traditional method (NPU = 35%; BV = 48%) and in extruded flat bread with 5% sugar (NPU = 35%; BV = 51%) were significantly reduced (P less than 0.05). In extruded flatbread without and with 2.5% sugar, NPU and BV of the raw material were essentially maintained. The results indicate that in comparison with the baking process the extrusion of flatbread is more suitable to protect the protein quality. The addition of more than 2.5% sugar caused protein damage in extruded flatbread, presumably as a result of Maillard reactions.

Bread↗