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The elongating leaves of young rye seedlings do not show a gravitropic response when placed horizontally. However, after treatment with ethylene, either supplied exogenously via ethephon or by application of its precursor 1-aminocyclopropane-1-carboxylic acid (ACC), gravicompetence is seen. The inhibition of ethylene perception by 1-methylcyclopropene (MCP) prevents gravicompetence. Young rye leaves provide a useful model system in which to identify the components of the gravity sensing or response systems, the presence of which govern gravicompetence.
A recent study demonstrated that gravi-incompetent coleoptile-less seedlings of rye exhibit gravi-competence after exogenous application of ethylene. Treatments and conditions which induce and interfere with this phenomenon were analysed in more detail. Aminocyclopropane-1-carboxylic acid (ACC) as a precursor of ethylene has similar gravicompetence-inducing effects and also appropriate conditions of light, which strongly enhances ethylene synthesis. Both effects can be inhibited by the ethylene-perception blocking agent methylcyclopropene (MCP) or inhibitors of ethylene synthesis such as aminovinylglycine (AVG), indicating that light exerts its gravicompetence-generating effect via induced/enhanced ethylene synthesis. Gain in gravicompetence is accompanied by the induced/enhanced occurrence of calreticulin and lipoxygenase as detected by 2D-gels and Q-TOFF-analyses. Previously gravicompetent, light-grown coleoptile-less seedlings are characterized by gravi-incompetent growth during subsequent horizontal gravistimulation when perception of ethylene is inhibited by MCP. The results demonstrate that continuous perception of ethylene is an indispensable step, permanently required for the regulation of gravitropic growth in germinating primary shoots of rye, either within the process of graviperception and/or of the transduction of the gravi-signal.
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A single-strand-specific nuclease from rye germ (Rn nuclease I) was characterized as a tool for secondary and tertiary structure investigation of RNAs. To test the procedure, yeast tRNA(Phe) and tRNA(Asp) for which the tertiary structures are known, as well as the 3'-half of tRNA(Asp) were used as substrates. In tRNA(Phe) the nuclease introduced main primary cuts at positions U33 and A35 of the anticodon loop and G18 and G19 of the D loop. No primary cuts were observed within the double stranded stems. In tRNA(Asp) the main cuts occurred at positions U33, G34, U35, C36 of the anticodon loop and G18 and C20:1 positions in the D loop. No cuts were observed in the T loop in intact tRNA(Asp) but strong primary cleavages occurred at positions psi 55, C56, A57 within that loop in the absence of the tertiary interactions between T and D loops (use of 3'-half tRNA(Asp)). These results show that Rn nuclease I is specific for exposed single-stranded regions.
A single-strand-specific nuclease from rye germ (Rn nuclease I) was used for secondary and tertiary structure investigations of tRNAs with large variable arms (class II tRNAs). We have studied the structure in solution of two recently sequenced tRNA(Leu): yeast tRNA(Leu)(ncm5UmAA) and bovine tRNA(Leu)(XmAA) as well as yeast tRNA(Leu)(UAG), tRNA(Leu)(m5CAA) and tRNA(Ser)(IGA). The latter is the only tRNA with a long variable arm for which the secondary and tertiary structure has already been studied by use of chemical probes and computer modelling. The data obtained in this work showed that the general model of class II tRNAs proposed by others for tRNA(Ser) can be extended to tRNAs(Leu) as well. However interesting differences in the structure of tRNAs(Leu) versus tRNA(Ser)(IGA) were also noticed. The main difference was observed in the accessibility of the variable loops to nucleolytic attack of Rn nuclease I: variable loops of all studied tRNA(Leu) species were cut by Rn nuclease I, while that of yeast tRNA(Ser)(IGA) was not. This could be due to differences in stability of the variable arms and the lengths of their loops which are 3 and 4 nucleotides in tRNA(Ser)(IGA) and tRNAs(Leu) respectively.
We studied the interaction of ethidium bromide with rye and calf thymus chromatin. Both types of chromatin have the same dye accessibility, which is about 50% of that of DNA. From this result we conclude that the molecular structure of these two chromatins is similar. For rye, the extraction of H1 produces no change in the binding of ethidium bromide. The subsequent extraction of H2A and H2B produces a 14% increase in the binding, and the removal of H3 and H4, another 54% increase. At this stage, the number of binding sites is still less than that of DNA. This is presumably due to the presence of some tightly bound non-histones. Thus, the arginine-rich histones and the tightly bound non-histones are most responsible for limiting the binding of ethidium bromide to rye chromatin.
GrainGenes, http://www.graingenes.org, is the international database for the wheat, barley, rye and oat genomes. For these species it is the primary repository for information about genetic maps, mapping probes and primers, genes, alleles and QTLs. Documentation includes such data as primer sequences, polymorphism descriptions, genotype and trait scoring data, experimental protocols used, and photographs of marker polymorphisms, disease symptoms and mutant phenotypes. These data, curated with the help of many members of the research community, are integrated with sequence and bibliographic records selected from external databases and results of BLAST searches of the ESTs. Records are linked to corresponding records in other important databases, e.g. Gramene's EST homologies to rice BAC/PACs, TIGR's Gene Indices and GenBank. In addition to this information within the GrainGenes database itself, the GrainGenes homepage at http://wheat.pw.usda.gov provides many other community resources including publications (the annual newsletters for wheat, barley and oat, monographs and articles), individual datasets (mapping and QTL studies, polymorphism surveys, variety performance evaluations), specialized databases (Triticeae repeat sequences, EST unigene sets) and pages to facilitate coordination of cooperative research efforts in specific areas such as SNP development, EST-SSRs and taxonomy. The goal is to serve as a central point for obtaining and contributing information about the genetics and biology of these cereal crops.
1. Chlorophyllase [EC 3.1.1.14] was extracted from the acetone-dried powder of the chloroplasts of greened rye seedlings with 1% cholate, and purified 870-fold with a yield of about 30%. The purification procedure was composed of fractionations with acetone and ammonium sulfate, and hydrophobic chromatography on a phenyl-Sepharose CL-4B column. 2. The purified enzyme was pure as analyzed by molecular-sieve chromatography and isoelectric electrophoresis. It had an isoelectric point of 4.5 and a molecular weight of 39,000. 3. The purified enzyme was stable at pH 6-9 and 4 degrees C. At pH 7.5, it was stable in the presence and absence of 30% acetone. However, at 30 degrees C, it was not stable above a 10% concentration of acetone. 4. The purified enzyme hydrolyzed chlorophylls a and b from spinach into chlorophyllides a and b and phytols, respectively; and bacteriochlorophyll a from Rhodospirillum rubrum into bacteriochlorophyllide a and a derivative of phytol, possibly all-trans-geranylgeraniol. The hydrolysis rates were stimulated to their maxima in the presence of 30% acetone; maximum stimulation was about 50% with bacteriochlorophyll a and about 400% with chlorophyll a. 5. At pH 7.5 and 30 degrees C in the presence of 30% acetone, the Km values and specific activities were 12 microM and 480 nmol . min-1 . mg-1 for chlorophylls a, and 4 microM and 170 nmol . min-1 . mg-1 for R. rubrum bacteriochlorophyll a, respectively.
1. The chlorophyllase [EC 3.1.1.14] purified from greened rye seedlings hydrolyzed the bacteriochlorophyll isolated from Rhodospirillum rubrum, but not the pigment bound to the membrane of chromatophores or spheroplasts from the bacterium. 2. Acetone, if added at such concentrations that the bound bacteriochlorophyll would not be solubilized, enabled the enzyme to hydrolyze the bound pigment. The acetone concentrations required for half the maximum hydrolysis rates were 16% with chromatophores and 7% with spheroplasts. 3. The enzymic hydrolysis of the bound bacteriochlorophyll in the presence of acetone removed bacteriochlorophyllide from the membrane, leaving its esterifying alcohol, possibly all-trans-geranylgeraniol, in situ. 4. Washing of chromatophores with 30% acetone removed about 10% of the bound bacteriochlorophyll. The bound pigment remaining after washing was not hydrolyzed by the enzyme unless acetone was added. 5. It seems possible that light-harvesting bacteriochlorophyll was mostly, if not all, bound to the inner surface of chromatophores (the outer surface of spheroplasts), having its esterifying alcohol residue buried in the membrane and its porphyrin residue emerging from the membrane into the inside solution; thus, chlorophyllase could not make contact with the ester linkage between the esterifying alcohol and porphyrin moieties of the pigment unless the esterifying alcohol residue was partly exposed.
Compared to rye plants grown under normal conditions of mineral nutrients, those grown under deficient conditions of mineral nutrients were shown to have a high potential activity of the root vacuolar H(+)-pyrophosphatase (H(+)-PPiase), with a low level of PPi in roots. Immunoblot analysis suggested a qualitative change of the enzyme.
Two experiments were conducted, each with 24 White Pekin ducks, to determine the AME and TME content of five feedstuffs for ducks. In each experiment, fasting losses were obtained from six ducks and six ducks were used for each feedstuff. Each experiment lasted 102 h with an initial 48-h period and a 54-h excreta collection period. During the first 48-h period, all birds were tube-fed dextrose (30 g/100 mL of water) at 8 and 32 h after feed was withdrawn. Thirty grams of each feedstuff were tube-fed (30 g/100 mL of water) at 48 and 54 h after feed was withdrawn. The birds from which fasting losses were obtained were intubated with 30 g dextrose (30 g/100 mL of water) at 48 and 54 h after feed was withdrawn. Excreta were collected during the last 54 h into bags screwed onto lids sutured around the vent of each bird. In the first experiment, the feedstuffs evaluated were corn, dehulled oats, and wheat. The fasting energy and nitrogen losses per bird in the 54-h collection period were 12.1 kcal and 0.29 g, respectively. The AMEn values for the birds fed corn, dehulled oats, and wheat were 3.10, 3.48, and 3.14 kcal/g, respectively. The TMEn values for the respective feedstuffs were 3.27, 3.64, and 3.30 kcal/g. In the second experiment, the feedstuffs evaluated were corn, parboiled rice, and rye. The fasting energy and nitrogen losses per bird in the 54-h collection period were 18.9 kcal and 1.09 g, respectively. The AMEn values for the birds fed corn, parboiled rice, and rye were 3.24, 3.45, and 2.69 kcal/g, respectively. The TMEn values for the respective feedstuffs were 3.40, 3.61, and 2.85 kcal/g. The data provide new information on AMEn and TMEn values of corn, wheat, parboiled rice, dehulled oats, and rye for ducks.
A total of 640 22-wk-old pullets (Shaver SX 288) housed four birds per cage in 40 experimental units (four cages per unit), were randomly assigned eight experimental diets in a 2 x 2 x 2 factorial arrangement. The treatments consisting of two grain sources (wheat and rye) two levels each of crude enzyme preparation (0 and 0.1% Roxazyme G), and added inorganic phosphorus (0 and 0.105%) were fed for five 4-wk periods. At 42 wk of age, 40 individually caged layers were fed the experimental diets with 0.3% chromic oxide (5 individual birds per treatment) to determine AMEn and available P. Plasma P and Ca were also determined. Egg production, feed intake (FI), egg weight, feed efficiency (FE), and specific gravity of eggs were significantly (P < or = 0.05) affected by the experimental periods. Hens fed wheat-based diets had higher (P < or = 0.05) tibia ash (54.3 vs 52.5%), excreta dry matter (22.0 vs 17.7%), and eggshells with > or = 1.080 specific gravity (93.5 vs 89.9%) than birds fed rye. Enzyme supplementation significantly improved AMEn (P < or = 0.01) and FE by 6.2 and 3%, respectively. Egg production increased numerically from 87.6 to 90.1%. Inorganic P supplementation significantly increased egg production (P < or = 0.01), FI, FE, and AMEn (P < or = 0.05) by 4.4, 2, 3, and 2.8%, respectively, but significantly (P < or = 0.05) decreased the proportion of eggs having a specific gravity > or = 1.080 from 92.8 to 90.6%. The enzyme and inorganic P supplementation had no effect on tibia ash content and total plasma Ca and P. Rye can be used in layer rations yielding satisfactory performance when fed with a fungal crude enzyme preparation high in pentosanase/xylanase activity.
Two experiments were conducted to examine the effects of different fat types, i.e., soybean oil (S) and beef tallow (T), in rye-based broiler diets, either unsupplemented (-) or supplemented (+) with xylanase (Avizyme 1300 at 1 g/kg diet), on selected bacterial groups adhering to the epithelium of crop, duodenum, jejunum, and ileum (Experiment 1, 16 d of age), on mean retention time (MRT) of digesta, and on digestibility of N and dry matter in successive segments of the digestive tract (Experiment 2, 24 d of age). Live weight of enzyme-treated and S-fed chickens was significantly higher than that for unsupplemented or T-fed birds, respectively, in both experiments. In Experiment 1, a reduction in bacterial colonization from crop to duodenum was followed by a continuous increase as far as the ileum. Xylanase supplementation significantly reduced enterobacteria and total anaerobe microbes with a similar trend for Gram-positive cocci and enterococci. The latter two groups were significantly enhanced in birds fed T. In Experiment 2, xylanase supplementation resulted in a decrease in MRT in several segments of the digestive tract. This effect was most pronounced in the small intestine, where MRT of 268, 217, 241, and 209 min in groups S-, S+, T-, and T+, respectively, were measured. Apparent digestibility of N and dry matter was slightly improved by xylanase supplementation in the jejunum and ileum. Nitrogen digestibility by the terminal ileum was 80.3, 83.7, 79.4, and 82.2% for the S-, S+, T-, and T+ groups, respectively, and dry matter digestibility amounted to 61.2, 65.5, 62.1, and 64.0%, respectively.