Effect of genetic diversity in Hordeum vulgare L. and Secale cereale L. on their crossability and the development and viability of hybrid embryos and plants

A study was made of the effect of genetic diversity in cultivated barley and rye on seed setting, embryonic development, and viability of barley x rye hybrids grown in vitro and planted. The variation in seed-setting frequency was shown to be determined mainly by vegetation conditions and random factors, while the variation in proportion of seeds with embryos among the total seed set was determined by the genetic diversity of barley and rye. Selection of barley genotypes was shown to increase the yield of viable barley x rye hybrids. Based on the phenotypic analysis of barely x rye hybrids obtained in various combinations, low viability and hybrid lethality were attributed to the hybrid necrosis genes expressed from early ontogeny. These genes are assumed to be widespread among barley and rye varieties.     

Molecular and chromosomal organization of DNA sequences coding for the ribosomal RNAs in cereals

The chromosomal locations of ribosomal DNA in wheat, rye and barley have been determined by in situ hybridization using high specific activity ¹²⁵I-rRNA. The 18S-5.8S-26S rRNA gene repeat units in hexaploid wheat (cv. Chinese Spring) are on chromosomes 1B, 6B and 5D. In rye (cv. Imperial) the repeat units occur at a single site on chromosome 1R(E), while in barley (cv. Clipper) they are on both the chromosomes (6 and 7) which show secondary constrictions. In wheat and rye the major 5S RNA gene sites are close to the cytological secondary constrictions where the 18S-5.8S-26S repeating units are found, but in barley the site is on a chromosome not carrying the other rDNA sequences. — Restriction enzyme and R-loop analyses showed the 18S-5.8S-26S repeating units to be approximately 9.5 kb long in wheat, 9.0 kb in rye and barley to have two repeat lengths of 9.5 kb and 10 kb. Electron microscopic and restriction enzyme data suggest that the two barley forms may not be interpersed. Digestion with EcoR1 gave similar patterns in the three species, with a single site in the 26S gene. Bam H1 digestion detected heterogeneity in the spacer regions of the two different repeats in barley, while in rye and wheat heterogeneity was shown within the 26S coding sequence by an absence of an effective Bam H1 site in some repeat units. EcoR1 and Bam H1 restriction sites have been mapped in each species. — The repeat unit of the 5S RNA genes was approximately 0.5 kb in wheat and rye and heterogeneity was evident. The analysis of the 5S RNA genes emphasizes the homoeology between chromosomes 1B of wheat and 1R of rye since both have these genes in the same position relative to the secondary constriction. In barley we did not find a dominant monomer repeat unit for the 5S genes.     

Mechanical Transmission of Barley Mild Mosaic Virus (BaMMV) to Rye (Secale cereale L.)

After mechanical spraygun inoculation barley mild mosaic virus (BaMMV) was detected in barley cv. ‘Gerbel’ (control) as well as in rye cv. ‘Somro’, but not in wheat cv. ‘Kanzler’ and oat cv. ‘Alfred’. ELISA values of infected barley and rye were similar. Furthermore, infected rye plants developed symptoms typical for barley yellow mosaic virus infection.     

Earlier onset of DNA fragmentation in leaves of wheat compared to barley and rye

We have found extensive nucleosomal fragmentation of native DNA extracted from leaves of healthy cereal plants, as indicated by ladder patterns on agarose gels and TUNEL staining. The time of first appearance of fragmentation differed among cereals. Native DNA from the first leaf of 10-day-old plants formed a clear ladder pattern of multiples of 180 bp fragments in wheat and triticale but not in barley and oats. In one cultivar of rye a weak ladder pattern occurred but not in another. Freezing and thawing of samples before DNA extraction resulted in much more extensive DNA fragmentation in wheat but not in rye and barley, indicating that DNA-degrading enzymes are present in the cytoplasm of wheat, but not in barley and rye, at this stage. In barley, nucleosomal fragmentation was first detected in 25-day-old plants. These results indicate that programmed cell death takes place in developing leaves of young cereal plants, but that the time of onset differs among cereal species.     

Repeated sequence DNA relationships in four cereal genomes

The effect of DNA fragment size on the extent of hybridisation that occurs between repeated sequence DNAs from oats, barley, wheat and rye has been investigated. The extent of hybridisation is very dependent on fragment size, at least over the range of 200 to 1000 nucleotides. This is because only a fraction of each fragment forms duplex DNA during renaturation. From these results estimates of the proportions of repeated sequences of each of the cereal genomes that are homologous with repeated sequences in the other species have been determined and a phylogenetic tree of cereal evolution constructed on the basis of the repeated sequence DNA homologies. It is proposed that wheat and rye diverged after their common ancestor had diverged from the ancestor of barley. This was preceded by the divergence of the common ancestor of wheat, rye and barley and the ancestor of oats. Once introduced in Gramineae evolution most families of repeated sequences appear to have been maintained in all subsequently diverging species. — The repeated sequences of oats, barley, wheat and rye have been divided into Groups based upon their presence or absence in different species. Repeated sequences of related families are more closely related to one another within a species than between species. It is suggested that this is because repeated sequences have been involved in many rounds of amplification or quantitative change via unequal crossing over during species divergence in cereal evolution.     

Regeneration potential of different wheat, rye and barley species in leaf explant culture

Regeneration potential of different wheat, rye and barley species in leaf explant culture. Comparative analysis of the induction ability of morphogenetic processes in vitro has been carried out in 16 wheat genotypes, 4 barley species and 6 rye genotypes. It has been shown that tetra- and hexaploid wheat species as well as wild barley species exhibited the highest embryogenic potential in the leaf explant culture while diploid wheat species and rye genotypes showed the lowest one. Genotypic dependence of processes of callus formation, induction of embryogenic calli and regeneration was revealed in the studied species.     

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.

     

Identification of Bilby, a diverged centromeric Ty1-copia retrotransposon family from cereal rye (Secale cereale L.).

A diminutive rye chromosome (midget) in wheat was used as a model system to isolate a highly reiterated centromeric sequence from a rye chromosome. Fluorescence in situ hybridization (FISH) shows this sequence localized within all rye centromeres and no signal was detected on wheat chromosomes. DNA sequencing of the repetitive element has revealed the presence of some catalytic domains and signature motifs typical of retrotransposon genes and has been called the Bilby family, representing a diverged family of retrotransposon-like elements. Extensive DNA database searching revealed some sequence similarity to centromeric retrotransposons from wheat, barley, and centromeric repetitive sequences from rice. Very low levels of signal were observed when Bilby was used as a probe against barley, and no signal was detected with rice DNA during Southern hybridization. The abundance of Bilby in rye indicates that this family may have diverged from other distantly related centromeric retrotransposons or incorporated in the centromere but rapidly evolved in rye during speciation. The isolation of a rye retrotransposon also allowed the analysis of centromeric breakpoints in wheat-rye translocation lines. A quantitative analysis shows that the breakpoint in IDS.1RL and 1DL.1RS and recombinant lines containing proximal rye chromatin have a portion of the rye centromere that may contribute to the normal function of the centromeric region.     

Features of crossability, haploidy and polyembryony in hybrid combinations between common barley Hordeum vulgare L. (2n = 14) and wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya

The role of individual chromosomes of rye in the manifestation of crossability and seedling development in hybrid combinations between common barley Hordeum vulgare L., cultivar Nepolegayushchii (2n = 14) and five wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya (2n = 40 wheat + 2 rye chromosomes). Crossability, which was measured by two parameters--frequency of set grains and frequency of grains with embryos--was shown to be significantly affected by each of the five rye chromosomes examined: 1R, 2R, 3R, 5R, and 6R; the development of barley haploids was affected by rye chromosomes 1 R, 3R, and 5R. We were the first to demonstrate that polyembryony could be induced by mutual effects of barley cytoplasm and rye chromosome 1R. Possible mechanisms controlling the development of haploids and twins in hybrid combinations H. vulgare x T. aestivum/S. cereale are discussed. The conclusion is drawn that hybrid combinations between common barley and wheat-rye substitution lines can serve as new models for studying incompatibility mechanisms in distant crosses and genetic control of parthenogenesis.     

Performance of Growing-Finishing Pigs Fed Diets Based on Normal or Low Viscosity Rye Fed With and Without Enzyme Supplementation

One hundred and forty crossbred pigs (Pig Improvement Canada Ltd, Acme Alberta), weighing an average of 21.5 + 3.6kg, were assigned on the basis of sex, weight and litter to one of five dietary treatments. The control diet was barley-based, while in the remaining four treatments, the grain component consisted of either normal or low viscosity rye fed with or without pentosanase. Digestibility coefficients for dry matter, crude protein and gross energy were significantly ( P = 0.0001) higher for the rye-based diets than the barley based diet. There were no differences in digestibility coefficients between normal and low viscosity diets or between diets supplemented or unsupplemented with pentosanase. Over the entire experimental period (21.5 to 100.7kg), pigs fed the normal viscosity rye gained better ( P = 0.001) and had a higher daily intake ( P = 0.001) than pigs fed the low viscosity rye. Pigs fed the low viscosity rye tended to gain slower than pigs fed barley and with lower intake while pigs fed the normal viscosity rye had similar gains and intake to the pigs fed barley. Enzyme supplementation failed to improve pig performance. Males had higher gains and intake than females ( P < 0.05). Enzyme supplementation, rye viscosity or choice of cereal grain had no effect on any of the carcass traits measured. In conclusion, the results of the present experiment indicate that rye is a good alternative to barley for use in growing-finishing rations and becomes increasing attractive as the age of the pig increases. Breeding efforts directed towards reducing the viscosity of rye are unlikely to be successful in improving the nutritive value of rye for swine.     

On the origin of cultivated rye

The origin of cultivated rye has been studied, taking into consideration evidence from various fields. Based on morphological resemblances and cytogenetic affinities, cultivated rye is included in Secale cereale L. emend. Sencer, which also includes annual wild and weedy ryes. Wild populations of 5. cereale , which have evolved from Secale montanum Guss. emend. Sencer, invaded wheat and barley fields during the early days of cultivation and gave rise to weedy ryes with varying degrees of rachis brittleness. Cultivated rye was selected from weedy ryes for non-brittle rachis and bigger caryopsis both unconsciously and consciously by man. The geographic origin of cultivated rye is postulated for the Büyük Ari Dai (Mt. Ararat) and Lake Van area in eastern Turkey. It spread from this area as a weed in wheat and barley fields towards the north, east and west and imposed itself as a secondary crop under conditions unfavourable for wheat and barley. It thus became a crop in its own right in several places independently, in addition to it being known by the people living in the Caucasus and Transcaucasus from very early agricultural times.     

Resistance genes for rye stem rust (SrR) and barley powdery mildew (Mla) are located in syntenic regions on short arm of chromosome.

Genetic stocks were developed for the localization and eventual cloning of the stem rust resistance gene SrR that occurs in wheat lines carrying the 1RS translocation from Secale cereale 'Imperial' rye. We have used a mutation-based approach for molecular analysis of the SrR region in rye. Forty-one independent mutants resulting in loss of SrR resistance were isolated: many of these were deletions of various sizes that were used to locate SrR with respect to chromosome group 1S markers. The analysis of the mutants showed that markers about 1 Mb apart flanking the barley Mla locus also flank SrR. Additionally, three of the approximately 20 closely related sequences of Mla in rye are deleted in each of six interstitial deletion mutants of SrR. The results indicate that the SrR region in rye is syntenic to the Mla region in barley or that SrR is possibly orthologous to the Mla locus.     

SOME OBSERVATIONS ON THE CEREAL-ROOT EELWORM POPULATION OF FIELD PLOTS OF CEREALS WITH DIFFERENT SOWING TIMES AND FERTILIZER TREATMENTS

In the autumn of 1953 an experiment was begun to follow changes in the cereal-root eelworm population of small plots on a field in Shropshire. The plots were cropped with either oats, wheat, barley or rye, sown in the autumn and spring, and some plots had fertilizer. Each plot received the same treatment for 3 years; in the fourth year an indicator crop of spring oats was grown on all the plots.
Under rye and autumn-sown wheat the eelworm population fell to a level which permitted a good oat crop in 1957. Autumn-sown wheat, barley and rye generally produced lower eelworm populations than their spring-sown counterparts, but autumn-sown oats proved to be the most efficient host. The order of host efficiency was oats (best), barley, wheat, rye. The eelworm populations were generally higher on plots receiving fertilizer treatment. In this experiment all oat plots, and spring-sown barley plus fertilizer, produced populations which severely damaged the 1957 oat crop.     

Host range,mating type and population structure of Magnaporthe sp. of a single barley field in São Paulo state,Brazil

Brazil is blast disease hot spot because severe epidemics have occurred among wheat, triticale, rye, barley and oat crops. Although the first outbreak of barley blast appeared in 1998, little information is available. Therefore, this study aimed to examine host range, mating type composition and population structure of Magnaporthe sp. from a single barley field in São Paulo, Brazil. To examine pathogenicity, 25 Magnaporthe isolates were inoculated on five, three, two and two cultivars of barley, wheat, oat and rice, respectively, and one cultivar each of rye, corn, sorghum, triticale and certain weeds (Cenchrus echinatus, Setaria geniculata, Brachiaria plantaginea and Eleusine indica). Mating type distribution of 33 isolates was investigated by molecular tools. The genotypic divergence of 41 barley and five wheat isolates was investigated by 15 random amplified polymorphic DNA primers and unweighted pair group method with arithmetic mean. The host range of the barley blast pathogen included wheat, oat, rye and triticale but not rice and weeds. Sexual reproduction appeared to not be involved in the high genotypic diversity because only a single isolate, MAT1‐2, was identified. The majority of barley isolates clustered together with wheat blast, except for four, suggesting a different origin.     

Fluorescent labeling of plant chromosomes in suspension by FISH

By optimizing the concentration and time of treatment with hydroxyurea (HU), a DNA synthesis inhibitor, and trifluralin, a microtubule inhibitor, a highly effective (over 60%) cell cycle synchronization method for rye and barley meristem cells was developed. Chromosome suspensions containing highly purified and morphologically intact rye and barley chromosomes were prepared from the meristems of their root tips by homogenization. Digoxigenin-labeled 5S rDNA was used as a probe in FISH for the rye chromosomes in the suspension, and biotin-labeled 17S rDNA and centromeric DNA were used in FISH for the rye and barley chromosome suspensions, respectively. Bright signals were detected at the specific regions of interest on the chromosomes. The results indicate that the method developed in this study is useful for selection and sorting of chromosomes that are not distinguishable by other means, using specific fluorescent labeling by FISH of the chromosomes in suspension.     

Cytoplasmic DNA variation and relationships in cereal genomes

Summary Chloroplast (cp) and mitochondrial (mt) DNAs were isolated from four cereal genomes (cultivated wheat, rye, barley and oats) and compared by restriction nuclease analysis. Cleavage of cp and mt DNAs by Sal I, Kpn I, Xho I and EcoR I enzymes indicated that each cereal group contains specific cytoplasmic DNAs. A phylogenetic tree of cereal evolution has been obtained on the basis of cp DNA homologies. It is suggested that wheat and rye diverged after their common ancestor had diverged from the ancestor of barley. This was preceded by the divergence of the common ancestor of wheat, rye and barley and the ancestor of oats.The molecular weight of the different cp DNAs was determined from the Sal I and Kpn I patterns. cp DNAs from wheat, rye, barley and oats appeared to be characterized by a very similar molecular weight of about 80–82.10⁶ d.In the case of the mt DNAs, the great number of restriction fragments obtained with the restriction enzymes used prevented precise comparisons and determination of molecular weights.     

A Comparative Study of Early Seed Development in Genotypes of Barley and Rye

Early seed development was studied in 17 genotypes of barley,Hordeum vulgare L., and 11 genotypes of rye, Secale cerealeL. The numbers of cells and nuclei in the embryos and endospermsof developing seeds were scored daily for 5 days after selfpollination. For embryos, the mean cell doubling times variedfrom 9.2–12.9 h for barley and 15.7–22.7 h for rye.Endosperm mitotic cycle times of both species were shortestover the first 24 h after pollination but then became longer.A non-linear correlation was found between the number of embryocells and the number of endosperm nuclei in barely and rye andis similar to that for other members of the Triticeae. Hordeum vulgare L., Secale cereale L., barley, rye, embryo, endosperm, mean cell doubling time     

Endogenous rhythmicity of ethylene production in growing intact cereal seedlings

Ethylene evolution from etiolated barley (Hordeum vulgare), wheat (Triticum aestivum), and rye (Secale cereale) seedlings during coleoptile growth followed a rhythmic pattern, with a period of about 16 h for barley and wheat and 12 h for rye seedlings. Leaf emergence disturbed the established rhythm of ethylene evolution.     

Features of Resistance to Take-all Disease in Cereal Species Evaluated by Laboratory Assays

Development of take-all disease, caused by Gaeumannomyces graminis var. tritici, on wheat, barley and rye, respectively, susceptible, moderately susceptible and resistant under field conditions, was evaluated by laboratory assays. The root system, which grew either in vermiculite in plastic tubes or polyethylene sleeves, or in a sand mix in plastic pots, was inculated, and lesion frequency and extension, both considered as resistance factors, were monitored. In assays in tubes and pots, both enabling multiple infection points, the infection scores of wheat, barley and rye on a scale of 0–5 were in decreasing order. In assays in sleeves, with a single point inoculation, lesion length on barley and rye was similar and less than thaton, wheat, but percentage of infected roots was markedly lower on rye than on barley. For large-scale screening programs we suggest to employ both the Tube Assay, which is reliable and easy to perform and the Slanted Sleeve Assay, which is more sensitive allowing detection of even a small degree of resistance. Selected accessions could be subsequently evaluated by the Pot Assay, under more natural conditions, and the resistance to take-all must eventually be verified under field conditions.