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Metrafenone: studies on the mode of action of a novel cereal powdery mildew fungicide.

Powdery mildew fungi are among the major pathogens causing diseases of cereals in the world. The mode of action of a novel systemic benzophenone fungicide, metrafenone, which is based on a precursor that is discussed in the preceding paper, has been analysed on the powdery mildew fungi of barley (Blumeria graminis Speer f. sp. hordei Marchal) and wheat (Blumeria graminis Speer f. sp. tritici Marchal). Preventive treatments reduced germination and blocked development beyond formation of appressoria, which penetrated less often. Moreover, metrafenone turned out to be an efficient curative fungicide, which rapidly affected fungal survival at low concentrations. The fungicide induced swelling, bursting and collapse of hyphal tips, resulting in the release of globules of cytoplasm. Bifurcation of hyphal tips, secondary appressoria and hyperbranching were also frequently observed. A histochemical analysis showed that metrafenone caused disruption of the apical actin cap and apical vesicle transport as well as weakening of the cell wall at hyphal tips. Finally, metrafenone strongly reduced sporulation. Reduced sporulation was associated with malformation of conidiophores that showed irregular septation, multinucleate cells and delocalisation of actin. Microtubules appeared to be only secondarily affected in metrafenone-treated B. graminis. The results suggest that the mode of action of metrafenone interferes with hyphal morphogenesis, polarised hyphal growth and the establishment and maintenance of cell polarity. Metrafenone likely disturbs a pathway regulating organisation of the actin cytoskeleton.

Ascomycota↗

Variation in the effects of take-all disease on grain yield and quality of winter cereals in field experiments.

Relationships between take-all intensity and grain yield and quality were determined in field experiments on cereal crops using regression analyses, usually based on single-point disease assessments made during anthesis or grain-filling. Different amounts of take-all were achieved by different methods of applying inoculum artificially (to wheat only) or by using different cropping sequences (in wheat, triticale or barley) or sowing dates (wheat only) in crops with natural inoculum. Regressions of yield or thousand-grain weight on take-all intensity during grain filling were similar to those on accumulated disease (area under the disease progress curve) when these were compared in one of the wheat experiments. Regressions of yield on take-all intensity were more often significant in wheat than in the less susceptible crops, triticale and barley, even when a wide range of disease intensities was present in the latter crops. The regressions usually had most significance when there were plots in the severe disease category. Thousand-grain weight and hectolitre weight usually responded similarly to total grain yield. Decreased yield was often accompanied by a significant increase in the percentage of small grains. When severe take-all was present in wheat, regressions showed that nitrogen uptake was usually impaired. This was sometimes accompanied, however, by increased percentage nitrogen in the grain as a consequence of smaller grain size with decreased endosperm. Significant effects of take-all, both positive and negative, on Hagberg falling number in wheat sometimes occurred. Significant regressions of yield on take-all assessed earlier than usual, ie during booting rather than grain-filling in wheat and triticale and during anthesis/grain-filling rather than ripening in barley, had steeper slopes. This is consistent with observations that severe disease that develops early can be particularly damaging, whilst the crops, especially barley, can later express tolerance by producing additional, healthy roots. The regression parameters, including maximum potential yield (y-axis intercept) and the extrapolated maximum yield loss, also varied according to the different growing conditions, including experimental treatments and other husbandry operations. These differences must be considered when assessing the economic potential of a control measure such as fungicidal seed treatment.

Algorithms↗

Occurrence of barley yellow dwarf virus in autumn-sown cereal crops in the United Kingdom in relation to field characteristics.

The incidence of barley yellow dwarf virus and of its aphid vectors was surveyed in untreated parts of autumn-sown cereal crops, mainly wheat and barley, in the United Kingdom in 1995-8. The incidence of virus in the spring was related to the incidence of aphids in the preceding autumn. Both virus and aphid incidences could also be related to a range of crop and field characteristics, in particular sowing date, regions as defined by their geographical position, topography and climate, the proximity of the field to the sea, the extent of arable land in the vicinity of the field, and the aspect and size of the field. Proximity of cropped fields to setaside areas had no significant effect on either aphid or virus incidence.

Animals↗

Abnormalities of spindle and cytokine behavior leading to the formation of meiotic restitution nuclei in intergeneric cereal hybrids

Mobile stages of meiosis have been analysed by visualizing the spindle in fertile cereal F1 hybrids. We describe four different mechanisms of the formation of restitution nuclei in meiotic division: (1) centripetal migration of telophase chromosome groups from the poles of a curved spindle at early telophase; (2) centripetal migration of the chromosome groups at late telophase when cell plate formation has failed; (3) preferable migration of univalents to one of the poles although spindle appearance is morphologically normal; and (4) in the absence of chromosome segregation where kinetochore fibers have failed to form. Copyright 1999 Academic Press.

Journal Article↗

Satellite cereal yellow dwarf virus-RPV (satRPV) RNA requires a douXble hammerhead for self-cleavage and an alternative structure for replication.

The 110 nt hammerhead ribozyme in the satellite RNA of cereal yellow dwarf virus-RPV (satRPV RNA) folds into an alternative conformation that inhibits self-cleavage. This alternative structure comprises a pseudoknot with base-pairing between loop (L1) and a single-stranded bulge (L2a), which are located in hammerhead stems I and II, respectively. Mutations that disrupt this base-pairing, or otherwise cause the ribozyme to more closely resemble a canonical hammerhead, greatly increase self-cleavage. In a more natural multimeric sequence context containing the full-length satRPV RNA and two copies of the hammerhead, wild-type RNA cleaves much more efficiently than in the 110 nt context. Mutations in the upstream hammerhead, including a knock-out in the catalytic core, affect cleavage at the downstream cleavage site, indicating that multimers of satRPV RNA cleave via a double hammerhead. The double hammerhead includes base-pairing between two copies of the L1 sequence which extends stem I. Disruption of L1-L1 base-pairing slows cleavage of the multimer. L1-L2a base-pairing is required for efficient replication of satRPV RNA in oat protoplasts. Mutations that affect self-cleavage of the multimer do not correlate with replication efficiency, indicating that the ability to self-cleave is not a primary determinant of replication. We present a replication model in which multimeric satRPV RNA folds into alternative conformations that cannot form in the monomer. One potential metastable intermediate conformation involves L1-L2a base-pairing that may facilitate formation of the double hammerhead. However, we conclude that L1-L2a also performs some other essential function in the satRPV RNA replication cycle, because the L1-L2a base-pairing is more important than efficient self-cleavage for replication.

Avena↗

Cell fate specification in the cereal endosperm.

While superficially simple, endosperm development is a complex, dynamic process. Cereal endosperms contain three major cell types: starchy endosperm, transfer cells and aleurone. The localized accumulation of the END1 transcript in the syncitial endosperm suggests that signals from the maternal placental tissue specify transfer cell type early. Aleurone fate is plastic and requires the continual input of positional cues to maintain cell identity. Starchy endosperm appears to be the default cell type. Mutant patterns suggest that a regulatory hierarchy integrates endosperm development. Requirements for gametic imprinting, maternal : paternal genome ratios and putative chromatin modeling factors indicate the importance of genomic control.

Cell Differentiation↗

Nutritional evaluation of dry-roasted navy bean flour and mixtures with cereal proteins.

A navy bean flour prepared by dry roasting in a salt bed as a medium of heat exchange was found to have a higher PER than beans which had been autoclaved in the conventional manner. This difference was attributed to a small but significant improvement in the digestibility of the protein. The PER of the roasted beans was higher than the autoclaved beans even in the presence of supplemental methionine. When various proportions of roasted beans and corn were fed at a level of 8.3% protein in the diet, a mixture in which 40% to 60% of the protein was provided by either beans or corn had a PER essentially the same as casein. Diets containing roasted beans and various cereal grains (oats, barely, buckwheat, wheat germ, and rice) were formulated in proportions calculated to give the highest chemical scores. In most cases the PER' were not significantly different from that of casein, and, in the case of rice, the PER was higher than that of casein. Supplementation of such diets with their first limiting amino acid failed to produce a further enhancement of the PER.

Amino Acids, Essential↗

Nutritional evaluation of oilseeds and legumes as protein supplements to cereals.

Several oilseed and legume protein products were fed to rats as the sole source of dietary protein, and in blends with cereals for the determination of protein efficiency ratio (PER) and biological availability of amino acids. In addition oilseed protein isolates were fed to mice for the determination of PER. Results of the mouse study revealed that the adjusted PER (casein = 100)for Target rapeseed isolate (108) was higher than those of sunflower (74), safflower (77), soybean (86) or flax (92) isolates. Results of the rat trials revealed that the adjusted PER for Twoer rapeseed meal (88) was higher than those of fababean (21), field pea (59) and soybean meal (72). Supplementation with methionine (0.2%) resulted in improved PER for fababean (84), field pea (101) and soybean meal (97). Mustard flour and rapeseed flour gave PER of 109 and 106, respectively, while the value of sunflower flour was low (56). Protein isolates of Tower rapeseed and soybean gave PER of 92 and 80, respectively. Blending of legumes and oilseeds with wheat flour (PER = 28) gave high PER values (60--85), as also occurred in rice blends (71--88). Supplementation of wheat-legume blends with lysine (0.4%), methionine (0.2%) and threonine (0.1%) brought all PER values above 100. It appeared that differences in PER of the diets paralleled the levels of the first limiting amino acid for rat growth. Results of balance trials indicated that the availability of the limiting amino acid(s) was lower than other essential amino acids for each protein source.

Amino Acids↗

Effect on nitrogen retention by adults of different proportions of indispensable amino acids in isonitrogenous cereal-based based diets.

Nitrogen retention of adults who consumed diets in which cereals furnished 6.0 g N and 0.9 g of lysine was improved by increasing lysine to 1.8 g without altering other amino acids. In a second experiment, 70% of the 6.0 g of dietary N was supplied by rice + wheat and 30% by mixtures of amino acids so designed that the total intakes of amino acids were equivalent to those in diets containing 6.0 g N from whole egg (E), egg + potato (EP), rice + wheat (RW), rice + soy (RS), wheat + milk (WM) or corn + beans (CB). Mean N balances of young men in descending order were, g/day: E 0.69 +/- 0.23, RS 0.44 +/- 0.15, EP 0.43 +/- 0.09, WM 0.24 +/- 0.16, CB 0.16 +/- 0.13 and RW -0.02 +/- 0.10. In the same order, these diet provided, g/day: lysine 2.6, 1.9, 2.3, 1.6, 1.4 and 1.0; S-acids 2.2, 1.7, 1.6, 2.2, 1.9 and 2.3; and tryptophan 0.7, 0.4, 0.6, 0.5, 0.3 and 0.4. N balances resulting from diets E, RS and EP did not differ significantly from each other but E was superior to CB and RW (P less than 0.01). Relative amounts and propertions of the essential amino acids could be varied without altering nitrogen retention until at least one amino acid became limiting. Several patterns of indispensable amino acids therefore may be equally effective in meeting needs of adults, but both amounts and relative proportions should be considered.

Adult↗

Sugars act as signal molecules and osmotica to regulate the expression of alpha-amylase genes and metabolic activities in germinating cereal grains.

The molecular mechanisms that initiate and control the metabolic activities of seed germination are largely unknown. Sugars may play important roles in regulating such metabolic activities in addition to providing an essential carbon source for the growth of young seedlings and maintaining turgor pressure for the expansion of tissues during germination. To test this hypothesis, we investigated the physiological role of sugars in the regulation of alpha-amylase gene expression and carbohydrate metabolism in embryo and endosperm of germinating rice seeds. RNA gel blot analysis revealed that in the embryo and aleurone cells, expression of four alpha-amylase genes was differentially regulated by sugars via mechanisms beyond the well-known hormonal control mechanism. In the aleurone cells, expression of these alpha-amylase genes was regulated by gibberellins produced in the embryo and by osmotically active sugars. In the embryo, expression of two alpha-amylase genes and production of gibberellins were transient, and were probably induced by depletion of sugars in the embryo upon imbibition, and suppressed by sugars influx from the endosperm as germination proceeded. The deferential expression of the four alpha-amylase genes in the embryo and aleurone cells was probably due to their markedly different sensitivities to changes in tissue sugar levels. Our study supports a model in which sugars regulate the expression of alpha-amylase genes in a tissue-specific manner: via a feedback control mechanism in the embryo and via an osmotic control mechanism in the aleurone cells. An interactive loop among sugars, gibberellins, and alpha-amylase genes in the germinating cereal grain is proposed.

Carbohydrates↗

Polymorphism for ribosomal RNA gene arrangement in the mitochondrial genome of fall rye (Secale cereale L.).

A restriction-fragment-length polymorphism (RFLP) in mitochondrial DNA (mtDNA) was detected between varieties of fall rye (Secale cereale L.) by Southern hybridization with rrn18, the gene encoding the mitochondrial 18S ribosomal RNA. Restriction mapping showed that the RFLP is based on differing numbers of genomic contexts (one vs three) for a recombining-repeat element (the "18S/5S repeat"). From examination of other Secale species, we conclude that the one-context state arose relatively recently, putatively by deletion of two of an ancestral set of three distinct genomic loci containing the mitochondrial 18S/5S repeat. This is consistent with our earlier conclusion that the 18S/5S repeat has probably existed in at least two genomic copies throughout much of the history of the grass family (at least 40 million years). Interestingly, the intervarietal difference in the number of distinct rrn18 loci is not accompanied by a major difference in the number of rrn18 copies per unit mass of mtDNA. This suggests the existence of a mechanism that can compensate rather precisely for differences in mitochondrial gene dosage, perhaps by over-replication or stabilization of specific subgenomic molecules.

Base Sequence↗

Lactic acid fermentation in the production of foods from vegetables, cereals and legumes.

Lactic acid bacteria perform an essential role in the preservation and production of wholesome foods. Generally the lactic acid fermentations are low-cost and often little or no heat is required in their preparation. Thus, they are fuel-efficient. Lactic acid fermented foods have an important role in feeding the world's population on every continent today. As world population rises, lactic acid fermentation is expected to become even more important in preserving fresh vegetables, fruits, cereals and legumes for feeding humanity.

Dairy Products↗

Organization of retro-element and stem-loop repeat families in the genomes and nuclei of cereals.

Sequences homologous to the retro-element BIS-1 and the stem-loop repeat Hi-10 are present in the genomes of a number of cereal species. A detailed characterization of these elements indicated that they are non-randomly organized in the genomes of at least two of these species, namely barley and rye. In contrast to the BIS-1 retro-elements, the stem-loop repeats are also non-randomly organized into discrete domains in interphase nuclei from barley and rye. Features of the organization of these repeats along chromosomes and within interphase nuclei of rye, barley and rice are discussed.

Base Sequence↗

A DNA-polymerase-related reading frame (pol-r) in the mtDNA of Secale cereale.

Mitochondrial (mt)DNA of Secale cereale contains an open reading frame (pol-r), the potential translation product of which shows significant homology to the type-B DNA polymerase encoded by the S1 plasmid of Zea mays; it contains the highly-conserved domains IIa to V of family B polymerases. The pol-r ORF is transcribed, as proven by RT-PCR, but the transcript is not edited. Upstream of the putative start codon a potential promoter motif was detected, fitting well into the postulated consensus sequence of the transcription initiation regions of Z. mays and Triticum aestivum. The pol-r ORF occurs in mtDNA of the fertile rye variety "Halo" and the cytoplasmic male-sterile (CMS) line "Pampa". Both ORFs are almost identical, apart from the 3' terminus; pol-r from Halo can code for 289 amino acids, pol-r from Pampa for 312 amino acids. Based on codon usage and the lack of editing, pol-r is considered to be a "young" gene, probably introduced in the mtDNA of rye by recombination with an mt plasmid.

Amino Acid Sequence↗

[Thorium in soil, vegetables, cereals and fruit].

Thorium contents (alpha-activities of the naturally occurring isotopes Th-228, Th-230, and Th-232) were determined in soils, vegetables, cereals, and fruits. The thorium content of plants depends on the degree of contamination by soil resuspension and thus on the specific surface of the plants. The activity of the isotope Th-230 is almost the same as that of the main isotope Th-232. Th-228, with about the same activity as Th-232 in soil, increases to about 10-fold the activity in vegetables, 29-fold in sweet chestnuts and 740-fold in Brazil nuts. Thorium concentration factors from the soil to these vegetable products are calculated; they include the total concentration, not only the soluble portion of thorium.

Edible Grain↗

[Residue analysis of pyrethroid insecticides in cereal grains, milled fractions and bread].

A procedure is described for the residual analysis of eight pyrethroid insecticides at levels of 2-5 micrograms/kg. Residues are extracted from cereal grains, flour or bread with a mixture of acetonitrile/water (2 + 1) and are partitioned into petroleum ether. Clean-up is performed by gel permeation chromatography on Bio-Beads S-X3 and an additional Florisil column. Electron capture gas liquid chromatography on a non-polar SE-30 capillary column separates the isomers of the compounds. Under the conditions used in the experiments with cyfluthrin, cypermethrin, deltamethrin, fenvalerate, and permethrin, the compound levels in rye and wheat did not decrease significantly. Losses during milling and baking were small. Consequently, pyrethroid insecticides remain effective in grain stored over a long period of time. However, considerable residues result in the milled fractions as well as in different types of bread.

Bread↗

A survey on the occurrence of Fusarium mycotoxins in Bavarian cereals from the 1987 harvest.

Bavarian cereals and wheat flour from the 1987 harvest were analysed for nivalenol (NIV) and deoxynivalenol (DON) using high performance liquid chromatography (HPLC) and for T-2 toxin and zearalenone (ZEA) by enzyme-linked-immunosorbent assay (ELISA). The study included 190 field samples of wheat, barley, rye and oat with visibly damaged ears, 45 samples of wheat intended for feed production and two series of wheat flour (type 550) and whole wheat flour collected in October 1987 and June 1988. The field samples examined showed a high DON contamination of wheat (87%) with an average of 3.96 mg/kg and a maximum of 43.8 mg/kg. Mean levels between 0.33 mg/kg and 0.27 mg/kg DON could be detected in barley, rye and oat. Of the wheat samples, 58% contained ZEA with a maximum of 1.560 mg/kg. The highest levels of ZEA were detected in samples which also showed high concentrations of DON. The NIV and T-2 toxin levels were comparatively low. Thirty percent of the samples showed NIV concentrations between 0.04 mg/kg and 0.29 mg/kg and 38% contained between 0.005 and 0.60 mg/kg of T-2. In the wheat samples for feed production, only DON was detected with an average of 0.190 mg/kg and a maximum of 0.75 mg/kg. The highest DON levels (0.58 mg/kg) from October 1987 were found in the wheat flour samples which were lower than the highest DON concentration (3.24 mg/kg) detected in the samples collected during June 1988. This fact was probably due to a substantial amount of non-contaminated wheat from 1986. The toxin concentrations in the whole wheat flour were not higher than in the type 550 flour.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

[Comparative studies of partial amino acid sequences of prolamins and glutelins from various cereals. VI. Anion exchange chromatography of peptide fractions obtained by cation exchange chromatography].

The main peptide fractions obtained from prolamins and glutelins of wheat, rye, barley, and corn by chymotryptic hydrolysis, gel filtration and cation exchange chromatography [1, 2] were further separated by anion exchange chromatography. The amino-acid compositions of the peptides obtained from wheat, rye and barley prolamins are closely related. Typical compositions are Glx greater than 44, Pro greater than 28, and Phe greater than 7 mol-%. The peptide fractions from corn prolamine are different (Glx greater than 20, Leu greater than 20, Ala greater than 12, Pro greater than 10 mol-%). The peptide fractions from wheat, rye and barley glutelins are less similar. Typical compositions are Glx greater than 30, Pro greater than 15, and Gly greater than 8 mol-%. Most of the fractions from corn glutelin contain many amino-acids with Glx, Leu, and Ala predominating. Wheat differs significantly from the other cereals because of the unique composition of some high molecular weight peptide fractions from glutelin, which are rich in Glx (greater than 49) and Gly (greater than 19 mol-%).

Amino Acid Sequence↗