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Studies on rye (Secale cereale L.) lines exhibiting a range of extract viscosities. 2. Rheological and baking characteristics of rye and rye/wheat blends and feeding value for chicks of wholemeals and breads.

Five rye lines exhibiting a wide range of extract viscosities were evaluated for the rheological and baking properties of their flours, individually and in blends with hard red spring wheat flour. Commercial cultivars of rye and triticale were included in the study as controls. Extract viscosities of rye flours were higher than those of corresponding wholemeals, indicating shifting of water-extractable arabinoxylan into flour during roller milling. Falling numbers of the rye flours correlated positively with their extract viscosities in the presence (r = 0.73, p < 0.05) or absence (r = 0.65, p < 0.05) of an enzyme inhibitor. Farinograms revealed the weakness of rye and triticale flours compared to wheat flour. Extract viscosities of rye flours were negatively correlated (r = -0.65, p < 0.05) with mixing tolerance index and positively correlated (r = 0.64, p < 0.05) with dough stability, suggesting a positive impact of extract viscosity on dough strength. Extract viscosity was negatively correlated (r = -0.74, p < 0.05) with loaf volume and specific volume (r = -0.73, p < 0.05) and positively correlated (r = 0.73, p < 0.05) with loaf weight of rye/wheat bread. Overall, the results indicated that 30% of flour from high or low extract viscosity rye could be incorporated into rye/wheat breads without seriously compromising bread quality. Inclusion of rye, particularly high extract viscosity rye, in chick diets seriously impeded growth performance and feed efficiency. Part of the arabinoxylan survived bread-making and exerted an effect on chicks, although substantially lower digesta viscosities were observed in chicks fed rye bread diets than in those fed rye wholemeals.

Animal Feed↗

Antioxidant effects of phenolic rye (Secale cereale L.) extracts, monomeric hydroxycinnamates, and ferulic acid dehydrodimers on human low-density lipoproteins.

Dietary antioxidants that protect low-density lipoprotein (LDL) from oxidation may help to prevent atherosclerosis and coronary heart disease. The antioxidant activities of purified monomeric and dimeric hydroxycinnamates and of phenolic extracts from rye (whole grain, bran, and flour) were investigated using an in vitro copper-catalyzed human LDL oxidation assay. The most abundant ferulic acid dehydrodimer (diFA) found in rye, 8-O-4-diFA, was a slightly better antioxidant than ferulic acid and p-coumaric acid. The antioxidant activity of the 8-5-diFA was comparable to that of ferulic acid, but neither 5-5-diFA nor 8-5-benzofuran-diFA inhibited LDL oxidation when added at 10-40 microM. The antioxidant activity of the monomeric hydroxycinnamates decreased in the following order: caffeic acid > sinapic acid > ferulic acid > p-coumaric acid. The antioxidant activity of rye extracts was significantly correlated with their total content of monomeric and dimeric hydroxycinnamates, and the rye bran extract was the most potent. The data suggest that especially rye bran provides a source of dietary phenolic antioxidants that may have potential health effects.

Antioxidants↗

Grain characteristics, chemical composition, and functional properties of rye (Secale cereale L.) as influenced by genotype and harvest year.

Grain characteristic, chemical composition, and functional properties of rye were measured in 19 different cultivars grown in one location in up to 3 years. The cultivars included 8 adapted hybrids, 7 adapted population cultivars, and 4 nonadapted population cultivars. The results showed a significant influence of both harvest year and genotype on grain characteristics, chemical composition, and functional properties of the grain. Multivariate data analysis confirmed that the variations in the data were explained by yearly and genotype differences. Calculations of variance components showed that the variations in plant height, harvest yield, and protein content were mainly due to genotype differences and to a lesser extent to differences among harvest years. The kernel weight, hardness index, and content of dietary fiber components, however, were more strongly influenced by the harvest year than by the genotype. Differences in starch properties measured by falling number (FN), amylograph peak viscosity, and temperature at peak viscosity were more strongly influenced by harvest year. The water absorption was strongly influenced by genotype effects, compared to yearly differences. FN and amylograph peak temperature were positively correlated (r = 0.94). No correlation was found between the water absorption and the relative proportion of water-extractable arabinoxylan (AX) compared to the total AX content. However, the degree of ferulic acid cross-linking showed a negative correlation (r = -0.70) with the water absorption.

Analysis of Variance↗

Cultivar and year-to-year variation of phytosterol content in rye (Secale cereale L.).

Intake of phytosterols (and -stanols) has been shown to decrease the level of low-density lipoprotein cholesterol and thus protect against development of cardiovascular diseases. Therefore, studies on the cultivar and year-to-year variation in phytosterol content in rye grains have been performed. The phytosterol content and composition of different rye cultivars, grown under identical conditions on the same field in three consecutive years, were analyzed. Both cultivar and year-to-year variation in sterol content were statistically significant (p < 0.0001). The total sterol content varied from 1007 +/- 21 mg/kg in the highest yielding cultivar, Tsulpan 3, to 761 +/- 10 mg/kg in the lowest yielding cultivar (Amando in the 1999 harvest). Because the meteorological conditions varied substantially between the different years, it was possible to deduce the impact of varying weather conditions on phytosterol content in the different cultivars. The studied cultivars had all the lowest phytosterol contents in the dry and warm harvest season of 1999. Although there were statistically significant cultivar and year-to-year variations in the sterol composition (p < 0.0001), these were only between 2 and 4% of the total sterol content.

Climate↗

Chromosomal location of 46 new RAPD markers in rye (Secale cereale L.).

The polymerase chain reaction (PCR) was used to locate RAPD markers using disomic wheat-rye addition lines in order to develop a set of molecular markers distributed on the seven rye chromosomes. We carried out RAPD amplifications on genomic DNA of wheat 'Chinese Spring' (CS), rye 'Imperial' (I), the amphiploid wheat-rye and the seven disomic wheat-rye addition lines (1R-7R) using 140 different 10-mer oligonucleotides. Forty six new RAPD markers were located on the seven rye chromosomes and all the disomic wheat-rye addition lines were identified on the basis of their amplification patterns. The number of RAPD bands located on 1R, 2R, 3R, 4R, 5R, 6R and 7R chromosomes were 5, 8, 11, 8, 8, 10 and 6, respectively. The seven wheat-rye addition lines can be distinguished using only the following three 10-mer oligonucleotides: OPA16, OPF19 and GEN3-605, the other RAPD primers being useful for this purpose. The use of these RAPDs as a source of molecular markers that could be linked to interesting genes or other important agronomic traits is discussed.

Chromosome Mapping↗

Generation of rye (Secale cereale L.) plants with low transgene copy number after biolistic gene transfer and production of instantly marker-free transgenic rye.

Rye is extremely recalcitrant to tissue culture and genetic transformation. We describe the efficient and reproducible production of stably expressing transgenic rye plants after biolistic gene transfer to callus tissue derived from immature embryos. Key factors in the production of transgenic rye plants include the identification of biolistic gene transfer parameters and a selection protocol, which does not affect its regeneration ability. The bar gene was used as a selectable marker and selection was performed by spraying the regenerated shoots with 0.05% Basta solution without any previous selection of tissue cultures. Based on Southern blot analysis, a total of 21 transgenic rye plants with independent transgene integration patterns were produced. A low transgene copy number was observed in most transgenic plants and 40% of the plants had a single transgene copy insert. The high frequency of single transgene copy inserts might be a consequence of the selection system, which is based on the identification of stably expressing transgenic plantlets rather than stably expressing tissue cultures. All transgenic rye lines with single transgene inserts showed stable transgene expression in sexual progenies, but indications of transcriptional and post-transcriptional gene silencing were observed in few transgenic lines with multiple transgene inserts. Tissue culture-based selection was not necessary for the generation of transgenic rye. The identification of 17 transgenic rye plants without using any selectable marker gene by PCR amplification of transgene sequences is also demonstrated. Instant generation of selectable marker-free transgenic rye avoids a negative impact of selective agents on the transgenic tissue cultures, responds to public concerns on the safety of selectable markers and will support multiple transformation cycles for transgene pyramiding.

Aminobutyrates↗

Flow karyotypes and chromosomal DNA contents of genus Triticum species and rye (Secale cereale).

The flow cytometry and chromosome imaging method were jointly used for analyzing genome content and chromosomal DNA content of hexaploid wheat (AABBDD), hexaploid triticale (AABBRR), tetraploid wheat (AABB), and AA, BB, DD genome donors and RR genome rye. Their genome sizes were 34.4 pg, 40.9 pg, 26.2 pg, 12.1 pg, 13.7 pg, 10.5 pg, and 16.9 pg, respectively. The 2C nuclear DNA content of BB genome donor with 13.7 pg was the highest value among the other genome donors, AA or DD. The genome content of tetraploid wheat, unlike hexaploid wheat or hexaploid triticale, was larger than the sum of the genomes of AA and BB genome donors. The DNA content of each chromosome ranged from 1.22 pg in DD genome donor to 2.61 pg in rye. Each chromosome peak was divided into three to four groups. Only one chromosome was included in the highest chromosomal DNA peak in hexaploid wheat, tetraploid wheat, DD genome donor and rye but two chromosomes in AA, BB genome donors, and hexaploid triticale. Correlation between 2C nuclear DNA content and chromosome density volume was the highest value compared with the other chromosomal parameters of chromosome area, or chromosome length.

Chromosomes, Plant↗

Growth inhibition and root ultrastructure of cucumber seedlings exposed to allelochemicals from rye (Secale cereale).

Inhibition of "Calypso" cucumber seedling growth by rye allelochemicals, 2(3H)-benzoxazolinone BOA and 2,4-dihydroxy-1,4(2H)-benzoxazin-3-one DIBOA, was studied by analyzing the growth of seedling tissues and organs. Light and electron microscopy of seedling root cells were also carried out to investigate the mechanism(s) of root growth inhibition and mode of action of these compounds. BOA inhibited root elongation and reduced the number of cucumber lateral roots by 77 and 100% at 0.1 and 0.43 mg BOA/ml deionized (DI) water, respectively. DIBOA also inhibited root growth, but did not affect the number of lateral roots. BOA increased size of cucumber cortical root cells fivefold, but DIBOA had no effect. Both compounds reduced the regeneration of root cap cells and increased the width of cortical cells resulting in increased root diameter. BOA and DIBOA caused increased cytoplasmic vacuolation, reduced ribosome density and dictyosomes, reduced number of mitochondria, and reduced lipid catabolism. Starch granules in amyloplasts of seedling roots treated with BOA and DIBOA were also greatly reduced compared to the control. Changes in cellular ultrastructure indicated that BOA and DIBOA reduced root growth by disrupting lipid metabolism, reducing protein synthesis, and reducing transport or secretory capabilities.

Benzoxazines↗

Early ribonucleic acid synthesis during the germination of rye (Secale cereale) embryos and the relationship to early protein synthesis.

Incorporation studies with radioactive precursors showed that synthesis of protein and RNA is initiated in germinating embryos of rye within the first hour of imbibition of water. By polyacrylamide-gel fractionations of radioactive nucleic acid components, the appearance of products of transcription of the genome was shown to follow the sequence: heterogeneous (ribonuclease-sensitive) RNA, 4S and 5S RNA by 20min, 31S and 25S rRNA by 40min, and 18S RNA by 60min. "Fingerprint' analysis of T1-ribonuclease digests show that all the large oligonucleotides present in 25S and 18S RNA are present in the 31S species, indicating that 31S RNA is the precursor rRNA molecule to both 25S and 18S RNA. The importance of these early RNA syntheses and in particular the possible template function of the heterogeneous RNA is discussed in relation to the concept of long-lived mRNA and the coding for protein synthesis in the first hours of germination.

Edible Grain↗

Identification of S-locus linked PCR fragments in rye (Secale cereale L.) by denaturing gradient gel electrophoresis.

Rye inbred lines segregating at the S-locus and homozygous at the Z-locus were investigated by PCR with primers derived from Brassica SLG-sequences. After denaturing gradient gel electrophoresis (DGGE), a 280 bp PCR-fragment displays a polymorphism perfectly correlated to the underlying S-genotypes. This is the first report on S-related DNA polymorphism in a bifactorial self-incompatibility system of the Poaceae.

Alleles↗

Chromosome differentiation and pairing behavior of polyploids: an assessment on preferential metaphase I associations in colchicine-induced autotetraploid hybrids within the genus Secale.

Preferential chromosome association at metaphase I has been analyzed and compared in autotetraploid cells obtained by colchicine treatment of hybrid diploid rye plants with different degrees of chromosomal divergence between homologs. The tendency to identical over homologous, but not identical, pairing preferences detected when homologous partners are contributed by less related parental lines indicates that chromosome differentiation may play an important role on preferential pairing behavior of polyploids. However, associations between more similar (identical) partners are not always favored, thus suggesting that additional factors must be considered. Other hypotheses for explaining pairing preferences in competitive situations are discussed. No clear relationship has been found between multivalent frequencies at metaphase I and chromosome differentiation between homologs or preferential pairing behavior. Therefore evolutionary divergences among related genomes should be carefully stated when evaluated from metaphase I configuration frequencies.

Chromosomes↗

Cel1, probably encoding a cellobiohydrolase lacking the substrate binding domain, is expressed in the initial infection phase of Claviceps purpurea on Secale cereale.

At the host-pathogen interface of hyphae penetrating host cell walls in the rye ovary, a lack of cellulase-gold labeling of beta-1, 4-glucan in host cell walls indicates that enzymatic degradation of cellulose might be an important factor during the infection of rye by Claviceps purpurea. Using cbh1 from Trichoderma reesei as a probe, a putative cellulase gene (cel1) was isolated from a genomic library of the C. purpurea strain T5. The coding region of 1,616 bp contains two introns and a putative signal peptidase cleavage site, leaving a coding capacity of 437 amino acids for the mature protein. The derived amino acid sequence shares significant homology with other fungal cellobiohydrolases and lacks the substrate binding domain. Expression analysis using reverse transcriptase-polymerase chain reaction (RT-PCR) shows that cel1 is induced during the first days of infection of rye by C. purpurea. It may be involved in the penetration and degradation of host cell walls by depolymerizing plant beta-1, 4-glucan and, therefore, play a role in the infection process.

Amino Acid Sequence↗

CPTF1, a CREB-like transcription factor, is involved in the oxidative stress response in the phytopathogen Claviceps purpurea and modulates ROS level in its host Secale cereale.

CPTF1, a transcription factor with significant homology to ATF/CREB bZIP factors, was identified during an expressed sequence tag (EST) analysis of in planta-expressed genes of the phytopathogen Claviceps purpurea. Using a gene-replacement approach, deletion mutants of cptf1 were created. Expression studies in axenic culture showed that the H2O2-inducible gene cpcat1 (encoding a secreted catalase) had a reduced basal expression level and no longer responded to oxidative stress in the delta cptf1 mutant. Biochemical analyses indicated that CPTF1 is a general regulator of catalase activity. Delta cptf1 mutants showed significantly reduced virulence on rye. Electron microscopical in situ localization revealed significant amounts of H2O2 in delta cptf1-infected rye epidermal tissues, indicating that the plant tissue displayed an oxidative burst-like reaction, an event not detected in wild-type infections. These data indicate that CPTF1 is involved not only in oxidative stress response in the fungus but also in modulation of the plant's defense reactions.

Amino Acid Sequence↗

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↗

Allergens from rye pollen (Secale cereale). II. Characterization and partial purification.

Rye pollen was incubated for 30 min and proteins extracted at this time were collected as extract A (EA). The same pollen grains were resuspended in buffer and incubated for 18.5 h. Proteins extracted in this period were designated extract B (EB). Both extracts were subfractionated by DEAE ion-exchange chromatography and allergen presence in peaks detected by the dot-immunobinding technique. The results reveal that unretained proteins (peaks 1 and 2) and proteins eluted at 0.2 M NaCl from extract B contain the highest proportion of allergens. SDS-PAGE of chromatographic peaks showed that peak 2 from extract B contains a highly purified 28 kDa band. On the skin of allergic patients this band gave a stronger positive prick test than for the crude extract.

Allergens↗