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PubMed2003

Analysis and sorting of rye (Secale cereale L.) chromosomes using flow cytometry.

Procedures for chromosome analysis and sorting using flow cytometry (flow cytogenetics) were developed for rye (Secale cereale L.). Suspensions of intact chromosomes were prepared by mechanical homogenization of synchronized root tips after mild fixation with formaldehyde. Histograms of relative fluorescence intensity obtained after the analysis of DAPI-stained chromosomes (flow karyotypes) were characterized and the chromosome content of the DNA peaks was determined. Chromosome 1R could be discriminated on a flow karyotype of S. cereale 'Imperial'. The remaining rye chromosomes (2R-7R) could be discriminated and sorted from individual wheat-rye addition lines. The analysis of lines with reconstructed karyotypes demonstrated a possibility of sorting translocation chromosomes. Supernumerary B chromosomes could be sorted from an experimental rye population and from S. cereale 'Adams'. Flow-sorted chromosomes were identified by fluorescence in situ hybridization (FISH) with probes for various DNA repeats. Large numbers of chromosomes of a single type sorted onto microscopic slides facilitated detection of rarely occurring chromosome variants by FISH with specific probes. PCR with chromosome-specific primers confirmed the identity of sorted fractions and indicated suitability of sorted chromosomes for physical mapping. The possibility to sort large numbers of chromosomes opens a way for the construction of large-insert chromosome-specific DNA libraries in rye.

Cell Separation↗
PubMed1994

Detection of rye chromosome 2R using the polymerase chain reaction and sequence-specific DNA primers.

Sequences derived from a rye gamma secalin gene were used as primers in polymerase chain reactions using DNA obtained from a series of wheat and triticale genetic stocks. A 473-bp fragment, the predicted size based on the distance between the selected primers, was found only in rye, triticales, and wheat lines carrying rye chromosome 2RS. Use of triticale lines with various wheat chromosome substitutions confirmed the chromosomal origin of the rye-specific marker. The presence of the 473-bp PCR product was always associated with the production of 75K secalins in grain samples. Thus, the primer sequences, and the clone of origin (pSC503), were both derived from the SEC-2 locus of rye chromosome 2RS.

Base Sequence↗
PubMed1995

Fluorescent in situ hybridization and C-banding analyses of highly repetitive DNA sequences in the heterochromatin of rye (Secale montanum Guss.) and wheat incorporating S. montanum chromosome segments.

The molecular characterization of C-banded regions of Secale montanum Guss. by means of in situ hybridization was performed in order to provide new information about their chromosome structure relative to cultivated rye, Secale cereale L. Accurate identification of individual chromosomes was achieved using simultaneous and (or) successive fluorescent in situ hybridization (FISH) and C-banding. FISH identification was performed using total rye DNA, three highly repetitive rye DNA sequences (pSc119.2, pSc74, and pSc34), and the ribosomal RNA probes pTa71 (18S, 5.8S, and 26S rDNA) and pTa794 (5S rDNA). FISH was also used to identify the chromosome segment involved in two spontaneous translocation lines recovered from a 'Chinese Spring'--S. montanum wheat-rye addition line. FISH analysis revealed the exact translocation breakpoints and allowed the identification of the transferred rye segments. The value of this type of analysis is discussed.

Chromosome Banding↗
PubMed1996

A PCR assay for detection of a 2RL.2BS wheat-rye chromosome translocation.

A 2RL.2BS wheat-rye translocation, present in the wheat germplasm line Hamlet, carries a gene for resistance to Hessian fly biotype L, one of the most virulent biotypes presently encountered in wheat production environments. Unlike several other wheat-rye chromosome translocations common in wheat breeding programs, 2RL lacks genes encoding storage proteins or other easily selected markers. Oligonucleotide primers synthesized from published sequences derived from the R173 family of moderately repetitive rye DNA were used in the DNA polymerase chain reaction (PCR) to identify specific markers for 2RL. The same primers, when used with DNA extracted from additional wheat-rye translocation lines of importance to the wheat breeding community, gave distinctive PCR products for each genotype. The single primer pair, PAWS5 and PAWS6, may, therefore, have wide applicability for the identification of wheat-rye chromosomal translocations presently encountered in wheat breeding populations.

Base Sequence↗
PubMed2004

Different numbers of rye B chromosomes induce identical compaction changes in distinct A chromosome domains.

In rye each B chromosome (B) represents 5.5% of the diploid A genome. Rye Bs have several nuclear to whole plant effects although they seem to bear no genes except for the ones that lead to their maintenance within a population. In this context, and considering that rye Bs are enriched in repetitive non-coding regions that build up heterochromatin (het), we investigated the influence of Bs on the organization of two chromatin fractions, namely the ribosomal DNA (facultative het) and satellite (non-het) domain of rye chromosome 1 by silver staining on root tip metaphase cells. The results show that rye Bs cause condensation both in the NOR and in the chromosome 1 satellite domain. Since the silver staining technique used is indicative of the transcriptional activity of the NORs, the condensation observed at those loci demonstrates that the rRNA gene arrays are down-regulated in the presence of Bs, regardless of their number per individual. Furthermore, the organizational changes of metaphase NORs find parallel with the interphase organization of ribosomal chromatin, since the frequency of cells with intranucleolar condensed rDNA regions increases drastically and nuclear matrix attachment pattern is altered in the presence of the Bs. Our results show an identical effect of the Bs on the organization of two distinct chromosome domains displaying a presence/absence dichotomy.

Chromosomal Instability↗
PubMed1984

Proliferation of mouse lymphocytes to rye grass pollen extract: effect of inflammatory exudate cells.

The ability of inflammatory peritoneal exudate cells to modify a murine lymphoproliferative response to rye grass pollen extract was investigated. Popliteal and mesenteric lymph node cells from Balb/c mice immunised with either aqueous rye or rye grass emulsified in complete Freund's adjuvant were co-cultured with 2 X 10(3)-10(5) macrophage or polymorphonuclear neutrophil (PMN)-enriched exudate cells. There was a dose-dependent increase in the response to rye grass antigen with increasing numbers of PMN. A similar enhancement was seen with up to 10(4) macrophages per culture; higher numbers were inhibitory. Lymph node cells from rye grass or ovalbumin-immune mice that had been challenged with a combination of sensitising antigen plus 10(7) PMN gave an increased antigen-specific response in vitro. Neither antigen alone, PMN nor a mixture of PMN plus heterologous antigen was active. Adoptive transfer of a similar number of peritoneal macrophages plus antigen was also without effect. These results are in accord with the suggestion that cells obtained from acute inflammatory lesions can directly influence the activity of immunocompetent lymphocytes.

Animals↗
PubMed2003

Lignan precursors from flaxseed or rye bran do not protect against the development of intestinal neoplasia in ApcMin mice.

Phytoestrogens, like isoflavonoids and lignans, have been postulated as possible colorectal cancer protective constituents. To investigate this hypothesis, two high-fiber sources rich in lignan precursors, i.e., rye bran and flaxseed, were tested for their ability to modulate intestinal tumor development in ApcMin mice. Test diets consisted of a control diet (a Western-style diet, adjusted for fiber and/or phytate content) supplemented with 5% flaxseed or 30% rye bran. Chemical analysis of diets and blood samples confirmed the enhanced systemic exposure of mice fed the test diets to the major lignan precursors, i.e., secoisolariciresinol and matairesinol. No statistically significant difference was observed in the incidence and multiplicity of small intestinal and colon tumors at terminal sacrifice between mice fed the control diet or the diet supplemented with 5% flaxseed. With the rye bran diet a statistically significant enhancement of the number of small intestinal tumors in female mice was observed. The number of colon tumors, however, was comparable between the control and rye bran-fed mice of either sex. Furthermore, no activating point mutations in the K-ras oncogene nor positive immunohistochemical staining for the p53 gene were observed in a set of 48 colon tumors. In conclusion, our results demonstrate that increased intake of lignan precursors from flaxseed or rye bran, administered in a Western-style diet, does not protect against intestinal tumor development in an appropriate animal model for intestinal neoplasia such as the ApcMin mice.

Animals↗
PubMed2005

Rye bran diet increases epithelial cell apoptosis and decreases epithelial cell volume in TRAMP (transgenic adenocarcinoma of the mouse prostate) tumors.

A rye bran diet has been suggested to retard prostate cancer growth and increase tumor cell apoptosis in animal models and in men with prostate cancer. Transgenic mice designed to develop prostate cancer (transgenic adenocarcinoma of the mouse prostate, TRAMP) were therefore treated with a rye bran diet. TRAMP mice were fed a rye bran (n = 13) or a control diet (n = 15) from 4 to 24 wk of age. Tumor histology, cell proliferation, and apoptosis were accessed using stereological and immunohistochemical techniques. In the dorsolateral prostate lobe in TRAMP mice, epithelial cell apoptosis was increased by 31%, and the volume fraction of epithelial cells was reduced by 20% (P < 0.05) in the rye bran-treated group compared with the control diet group. There were, however, no significant changes detected in lobe weights or tumor grade. In conclusion, a rye bran diet increases epithelial apoptosis and reduces the epithelial cell fraction in TRAMP tumors.

Adenocarcinoma↗
PubMed1976

Effect of ultraviolet light and oral vitamin D3 on rachitic chicks fed diets containing either corn or rye.

Three experiments using day-old chicks were conducted in battery brooders to further study the rye-vitamin D antagonism. Birds were fed a vitamin D3-free diet containing corn or rye and submitted to diverse treatments for the first 10 days. At this time the chicks were either continued on the same diet or changed to other grain-type diets. The effect of ultraviolet light exposure on the chicks and of a single oral dose of vitamin D3 was studied, and body weight gain and bone ash were determined after a one-week experimental period. Rachitic chicks on a corn diet responded significantly better than rye-fed chicks to a single oral dose of vitamin D3, based on bone ash of fat-free, dry tibia. This rachitogenic effect of rye was completely overcome by exposing the chicks to ultraviolet light or by water extraction or acid-autoclaving this grain. The results also demonstrate that the interference by rye does not persist after this grain is removed from the diet.

Administration, Oral↗
PubMed1980

Growth depression of chicks fed a crude rye extract containing pectic substances.

A crude extract with a high proportion of pectic substances was prepared from rye. This was fed to chicks in two studies to determine if rye pectic substances are responsible for the growth depression of chicks fed rye. The growth of chicks was significantly (P greater than .01) depressed when fed a diet containing this extract. Addition of penicillin to the diet improved the growth of chicks. Increasing the concentration of the extract in the diet produced an incremental depression in weight gain. The intestines of chicks fed this extract contained large quantities of gas that were reduced by penicillin supplementation. These results suggest that rye pectic substances may be responsible for the growth depression of chicks fed rye.

Animal Feed↗
PubMed1987

Antinutritional activity of a water-soluble pentosan-rich fraction from rye grain.

A series of experiments was designed to analyze the antinutritive compound(s) in rye that depress nutrient utilization; these were found to be mainly associated with the rye flour fraction. The bran fraction, however, more effectively depressed feed intake than the flour fraction. A starch-rich isolate, which contained most of the antinutritive activity from rye, as well as a protein-rich isolate devoid of activity, were prepared from the flour. Treating the starch-rich isolate of rye flour with .1 N NaOH stabilized the antinutritive factor. Centrifugation of the NaOH isolate produced four fractions. The supernatant fraction contained considerable quantities of water-soluble and highly viscous pentosans and was the only fraction that significantly depressed nutrient utilization. Increasing additions of the water soluble fraction to a control diet resulted in nearly proportionate decreases in the retention of dry matter and fat. An ethanol fractionation produced an ethanol-insoluble precipitate that contained a two-fold increased concentration of the water-soluble pentosans and demonstrated a two-fold increase in antinutritive activity. These results support the hypothesis that the rye antinutritional compounds are viscous pentosans.

Animal Nutritional Physiological Phenomena↗
PubMed1991

The effects of enzyme supplementation on the nutritive value of rye grain (Secale cereale) for the young broiler chick.

Two dose-response experiments utilizing differing enzyme concentrations were carried out to determine the level of a crude enzyme preparation (Trichoderma viride cellulase) required to obtain maximum growth performance in young broiler chicks fed a 60% rye diet. Enzyme concentrations utilized in these studies ranged from .05 to 12.8 g enzyme/kg diet. Enzyme supplementation at the 3.2 g/kg diet level yielded increases in feed consumption and weight gain of 71 and 193%, respectively; and the feed to gain ratio was improved by 43% compared with that for the unsupplemented rye control diet. As no further growth response was obtained by increasing the enzyme concentration, the enzyme level utilized in a subsequent study was 3.2 g/kg diet. A balance study using 2-wk-old broiler chicks was carried out to determine the effect of enzyme supplementation on the bioavailable energy (AMEn) and apparent protein digestibility (APD) of a 60% wheat diet replaced by rye in 10% increments. In the 60% rye diets, the enzyme addition increased the dietary AMEn and APD by 23 and 12%, respectively, compared with the unsupplemented diet. The AMEn and APD of the 60% wheat control diet were increased 9.7 and 4.6%, respectively, by enzyme addition. The data suggest that the AMEn of rye grain is increased by enzyme supplementation; however, alternate methodology will have to be employed to quantitatively determine the extent of these improvements.

Animal Feed↗
PubMed1992

The effect of enzyme supplementation on the apparent metabolizable energy and nutrient digestibilities of wheat, barley, oats, and rye for the young broiler chick.

The influence of enzyme supplementation on the bioavailable energy (AME(n)) and apparent digestibilities of lipid (ALD) and protein (APD) in young broiler chicks was examined for diets containing either wheat, hulled or hulless barley, naked oats, or spring rye. Dietary AME(n), APD, and ALD values were depressed (P less than or equal to .01) for all test grains (except hulled Bedford barley) as the inclusion rate of the grain replacing wheat increased. The antinutritives, beta-glucans (barley and oats) and pentosans (rye), had the most pronounced effect on ALD. The decreases in ALD were 43, 77, and 67% for chicks fed diets containing 70% Scout barley (hulless), Terra oats, and Gazelle rye, respectively, compared with those fed the control wheat diet. Enzyme supplementation increased (P less than or equal to .01) AME(n), APD, and ALD for all test cereals. The corresponding increases in the AME(n), of the enzyme-supplemented diets containing 70% HY320 wheat, Bedford barley, Scout barley, Terra oats, and Gazelle rye diets were 4, 7, 42, 33, and 14%, respectively, compared with their unsupplemented counterparts. Enzyme treatment also improved (P less than or equal to .01) weight gains and feed conversion efficiencies of chicks fed diets containing each of the cereals. Overall, the results demonstrate that the nutritive value of cereal grains such as wheat, barley, oats, and rye can be improved by the addition of crude fungal extracts to the diet of young chicks.

Animal Feed↗
PubMed2003

The influence of rye cytoplasm on meiotic stability of tetraploid Secalotriticum.

Chromosome pairing in tetraploid Secalotriticum was analysed. In the studied plants wheat chromosomes in PMCs during metaphase I showed a higher degree of pairing, in comparison to the rye genome. This is reflected in a very low frequency of univalents and a higher frequency of ring bivalents. The occurrence of wheat univalents was dependent on wheat mixogenome. In plants with an unstabilized fourth homoeologous group, a heteromorphic bivalent 4A-4B was observed in 39.9% of PMCs, whereas in plants with an unstabilized seventh homoeologous group, chromosome 7A-7B pairing was found in all analysed cells. Rye univalents were present in all plants studied. The highest mean frequency of univalents and rod bivalents, both in wheat and in rye genomes, were recorded in plants whose first homoeologous group contained chromosome 1A. The mean number of terminal chiasmata per chromosome amounted to 1.78 in the wheat genome and 1.36 in the rye genome. It may be concluded that the plasmagenes in Secalotriticum did not increase the meiotic stability of the rye genome and also did not stabilize plant fertility.

Chromosomes↗
PubMed2003

The chromosomal location of rye AFLP bands.

23 AFLP bands were assigned to different rye chromosomes by means of two different sets of wheat-rye addition lines. Only one AFLP band could be assigned to 4R, and no specific AFLPs were found on the 5R chromosome. Only one AFLP band was explicitly assigned to 4R, and no specific AFLPs were found on the 5R chromosome. At least seven co-migrating AFLPs showed the same chromosomal location in both sets of addition lines. A total of 22 AFLPs were assigned to chromosome 1R using wheat-rye substitution lines. Six of them have counterparts in one of the addition lines analyzed, but only four have the same chromosomal location. Six and four of the total AFLPs located using addition (23) and substitution (22) lines segregated in the mapping population DS2 x RXL10, but only six were simultaneously assigned to the same chromosome by both approaches. Although co-migrating AFLPs could be located on different rye chromosomes using addition and substitution lines, we believe that AFLPs can be useful as rye chromosome markers.

Chimera↗
PubMedDate not supplied

[Actinomycete complexes in the rhizosphere of winter rye on soddy podzolic soil].

The structure of actinomycete complexes in the rhizosphere of winter rye (Secale cereale L.) varieties originating from different ecosystems and geographical zones was studied in field experiments on soddy podzolic soil. In addition to streptomycetes, the complexes studied contained actinomycetes of the genera Micromonospora and Streptosporangium, represented at high occurrence and comparable abundance rates. The rhizosphere of most of the studied rye varieties was dominated by micromonosporas. The antifungal potential of mycelial prokaryotes associated with winter rye was assessed. The taxonomic and functional structure of actinomycete complexes was shown to be similar in rye varieties originating from the nonchernozem zone of Russia. However, the actinomycete complex proved to be much different in the rye variety that developed in the steppe zone of the lower Volga region.

Actinobacteria↗
PubMed2005

[Genetic collection of meiotic mutants of rye Secale cereale L].

Genetic collection of meiotic mutants of winter rye Secale cereale L. (2n = 14) was created. Mutations were detected in inbred F2 generations after self-fertilization of the F1 hybrids, obtained by individual crossing of rye plants (cultivar Vyatka) or weedy rye with plants from autofertile lines. The mutations cause partial or complete plant sterility and are maintained in collection in a heterozygous state. Genetic analysis accompanied by cytogenetic study of meiosis has revealed six mutation types. (1) Nonallelic asynaptic mutations sy1 and sy9 caused the formation of only axial chromosome elements in prophase and anaphase. The synaptonemal complexes (SCs) were absent, the formation of the chromosome "bouquet" was impaired, and all chromosomes were univalent in meiotic metaphase I in 96% (sy1) and 67% (sy2) of cells. (2) Weak asynaptic mutation sy3, which hindered complete termination of synapsis in prophase II. Subterminal asynaptic segments were always observed in the SC, and at least one pair of univalents was present in metaphase I, but the number of cells with univalents did not exceed 2%. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19, which caused partially nonhomologous synapsis: change in pairing partners and fold-back chromosome synapsis in prophase I. In metaphase I, the number of univalents varied and multivalents were observed. (4) Mutation mei6, which causes the formation of ultrastructural protrusions on the lateral SC elements, gaps and branching of these elements. (5) Allelic mutations mei8 and mei10, which caused irregular chromatin condensation along chromosomes in prophase I, sticking and fragmentation of chromosomes in metaphase I. (6) Allelic mutations mei5 and mei10, which caused chromosome hypercondensation, defects of the division spindle formation, and random arrest of cells at different meiotic stages. However, these mutations did not affect the formation of microspore envelopes even around the cells, whose development was blocked at prophase I. Analysis of cytological pictures of meiosis in double rye mutants reveled epistatic interaction in the mutation series sy9 > sy1 > sy3 > sy19, which reflects the order of switching these genes in the course of meiosis. The expression of genes sy2 and sy19 was shown to be controlled by modifier genes. Most meiotic mutations found in rye have analogs in other plant species.

Chromatin Assembly and Disassembly↗
PubMed2006

[Effects of sodium nitroprusside on mitochondrial function of rye and wheat root tip under aluminum stress].

Sodium nitroprusside (SNP) could ameliorate the inhibition effect of Al on root growth of rye (Secale cereale L. cv. King) and wheat (Triticum aestivum L. cv. Jinmai47). Respiratory rate, P/O, OPR (oxygenated phosphate rate), R(3) (oxygen consumption rate with ADP and substrate present), R(4) (oxygen consumption rate with substrate) and RCR (respiratory control ratio, R(3)/R(4)) of root tips from rye and wheat decreased, as well as the activities of H(+)-ATPase, H(+)-PPase, Na(+)-K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase, but they increased with SNP treatment. It showed that mitochondrial respiratory functions of root tips from rye and wheat were damaged, phosphorylation was un-coupled by Al, but that of rye was less than that of wheat. Rye has high Al-resistance ability than wheat. SNP is one of donor of NO, it is suggested that NO can ameliorate remarkably respiratory dysfunction resulted from Al stress, so that NO can ameliorate the inhibition effect of Al on plant growth.

Aluminum↗