The effects of ‘gibberellin-insensitive’ dwarfing alleles in wheat on grain weight and protein content

Summary Three series of near-isogenic wheat lines differing in dwarfing alleles, in the varietal backgrounds of Maris Huntsman, Maris Widgeon and Bersee, and the F₂ grain on intravarietal F₁ hybrids, produced with a chemical hybridising agent, were examined for grain size and protein content. Individual F₂ grains from Rht1/rht, Rht2/rht and Rht3/rht F₁ spikes were classified for Rht genotype by assaying embryo half grains in a gibberellic acid seedling response test, while the remaining half was used for protein determination. Mean grain weight and protein percentage were lower in all homozygous isogenic lines and the Rht/rht F₁ hybrids than in the respective tall lines, in an allele dose-dependent manner. In all the hybrids, the Rht genotype of individual F₂ grains, which segregated within the spikes of F₁ plants, had no significant effects on grain weight or protein. Consequently, the pleiotropic effects of the Rht alleles on these yield and quality components must be attributed to their presence in maternal plant tissues rather than in the endosperm or embryo tissues of individual grains.     

Distance-constraint approach to protein folding. I. Statistical analysis of protein conformations in terms of distances between residues

A statistical analysis of protein conformations in terms of the distance between residues, represented by their C atoms, is presented. We consider four factors that contribute to the determination of the distanced _i,i+k between a given pair ofith and(i+k)th residues in the native conformation of a globular protein: (1) the distancek along the chain, (2) the size of the protein, (3) the conformational states of theith to(i+k)th residues, and (4) the amino acid types of the and(i+k)th residues. In order to account for the dependence on the distancek along the chain, the statistics are taken for three ranges, viz., short, medium, and long ranges (k8; 9k20; andk21; respectively). In the statistics of short-range distances, a mean distanceD _k and its standard deviationS _k are calculated for each value ofk, with and without taking into account the conformational states of all residues fromi toi+k (factors 1 and 3). As an Appendix, the relations for converting from the distances between residues into other conformational parameters are discussed. In the statistics of long-range distances, a reduced distanced* _ij (the actual distance divided by the radius of gyration) is used to scale the data so that they become independent of protein size, and then a mean reduced distanceD _l (a, a) and its standard deviation _l (a, a) are calculated for each amino acid pair (a, a) (factors 2 and 4). The effect of the neighboring residues along the chain on the value of the distanced* _ij is explored by a linear regression analysis between the actual reduced distanced* _ij and the mean value over theD _l for all possible pairs of residues in the two segments of the (i–2)th to the (i+2)th and the (j–2)th to the (j+2)th residues. The effect is assessed in terms of the tangentA _l (a, a) of the calculated regression line for each amino acid pair (a, a). In the statistics of medium-range distances, only factors 1 and 4 are considered, to simplify the analysis. The scaled distanced _i,i+k =(d _i,i+k -D _k )/S _k is used to eliminate the dependence onk, the distance along the chain. The propertiesD _m (a, a), _m (a, a) andA _m (a, a) corresponding toD _l (a, a), _l (a, a), andA _l (a, a), and also calculated for each amino acid pair (a, a). The results are interpreted as follows: the smaller values ofD _l (a, a) andD _m (a, a) indicate a preference of the pair (a, a) for a contact (e.g., pairs between hydrophobic amino acids, and pairs of Cys with aromatic amino acids), and the larger values of these quantities indicate a preference for distant mutual location (e.g., pairs between strong hydrophilic amino acids); the smaller values of _l (a, a) and _m (a, a) indicate a strong preference for either contact or noncontact (e.g., pairs between hydrophobic amino acids, and pairs between strong hydrophobic and hydrophilic amino acids, respectively), and the larger values of these quantities indicate the ambivalent/neutral nature of the preference for contact and noncontact (e.g., pairs containing Ser or Thr); the smaller values ofA _l (a, a) andA _m (a, a) indicate that the distance of an (a, a) pair is determined independently of the amino acid character of the neighboring residues along the chain (e.g., some pairs of Cys or Met with other amino acids) and the larger values of these quantities indicare that such amino acid character contributes strongly to the determination of the distance (e.g., pairs containing Ser or Thr, and pairs between amino acids with small side chains). The difference between the statistics for the long- and medium-range distances is also discussed; the former reflect the difference between the hydrophobic and hydrophilic character of the residues, but the latter cannot be easily interpretable only in terms of hydrophobicity and hydrophilicity. The data analyzed here are used in the optimization of an object function to compute protein conformation in a subsequent paper.     

Cloning and sequencing of the hemA gene of Rhodobacter capsulatus and isolation of a δ-aminolevulinic acid-dependent mutant strain

Summary The Rhodobacter capsulatus hemA gene, coding for the enzyme -aminolevulinic acid synthase (ALAS), was isolated from a genome bank by hybridization with a hemT probe from Rhodobacter sphaeroides. Subcloning of the initial 3.9 kb HindIII fragment allowed the isolation of a 2.5 kb HindIII-BglII fragment which was able to complement the -aminolevulinic acid-requiring (ALA-requiring) Escherichia coli mutant SHSP19. DNA sequencing revealed an open reading frame coding for a protein with 401 amino acids which displayed similarity to the amino acid sequences of other known ALASs. However, no resemblance was seen to the HemA protein of E. coli K12. Based on the sequence data, an ALA-requiring mutant strain of R. capsulatus was constructed by site-directed insertion mutagenesis. Introduction of a plasmid, containing the hemA gene of R. capsulatus on the 3.9 kb HindIII fragment, restored ALA-independent growth of the mutant indicating that there is only one gene for ALA biosynthesis in R. capsulatus. Transfer of the R factor pRPS404 and hybridization analysis revealed that the ALAS gene is not located within the major photosynthetic gene cluster.Part of this research was presented at the Symposium on Molecular Biology of Membrane-Bound Complexes in Phototrophic Bacteria, Freiburg, FRG, 2–5 August 1989     

On the mode of antirepressor action in anti-immune cells

Summary The mode of antirepressor action in anti-immune cells was analysed in respect to its two main features, low lysogenization and high cell killing. By means of complementation experiments between and i434 in anti-immune cells, it was shown that the antirepressor no longer channels phages towards lysis in such cells if the genes which are needed for lysogenization are provided in trans by the heteroimmune phage i434. Since complementation could be demonstrated, it was possible to exclude that direct action of the antirepressor over repressor production is responsible for the feature under analysis. It was also shown that both int- and cII-product are required to improve lysogenization and to prevent high levels of killing.Recipient of an EMBO predoctoral fellowship.     

A simple mathematical modeling of the growth of microorganism colonies

The growth rates of aspergillus, fusarium, and penicillium microorganism colonies in Czapex Dox Agar as a feed material, under room conditions, are observed to be linear. This phenomenon is mathematically modeled and exactly predicted on the basis of the exponential growth assumption of a single microorganism. The approach allows an easy determination of the multiplication constant of a had microorganisms been allowed to grow freely microorganism, in given conditions.     

Conserved elements in the 3′ untranslated regions of c-fos and actin mRNAs

In this study, the nucleotide sequences of the 3 untranslated regions (UTR) of the mouse and human c-fos genes, and the rat and human -actin genes were examined. It is shown (i) that the 3 UTR of c-fos is highly conserved between mouse and man, (ii) that multiple copies of a 12 bp element occur, in clusters, in the 3 UTR both of c-fos and of -actin. This conserved 12 bp element is analogous to the putative repressor binding site previously identified (Renan,Bioscience Reports,5 (1985), 739–753). These findings provide additional support for the proposal that regulatory signals are located in the 3 UTR's of certain genes.     

The multiplication constant of a microorganism in a colony is normally reduced by irradiation but still remains as a characteristic constant-A new approach to determining irradiation pasteurization doses

This work is based on a previous observation and on a related mathematical modeling regarding the linear growth of a colony of microorganisms under given conditions. We had previously shown that the growth rate of the colony is merely proportional to the individual exponential multiplication constant, , of the microorganisms.Tiny colonies of penicillium are subjected to different doses of irradiation. The subsequent observation of the colonies' growth rate beautifully furnishes a measure of how the multiplication constant, , of the microorganism is affected by irradiation.The plot of with respect to the irradiation dose, shows a linear interdependence between the two quantities. The extrapolation of this plot easily yields the radiation pasteurization dose of the microorganisms in hand.     

Photoperiodism and thermoperiodism in the predatory mite Amblyseius potentillae are probably based on the same mechanism

Summary A comparative study was made of the photoperiodic and thermoperiodic induction of diapause in the phytoseiid mite Amblyseius potentillae. Sensitivity to thermoperiod was found to be highest during the protonymphal and deutonymphal stages, with some sensitivity still being present in the young adult. Summation of both photoperiodic and thermoperiodic cycles was shown to take place, which demonstrated the presence of a photoperiodic counter as well as a thermoperiodic counter in these mites. Vitamin A appeared to be necessary for some early step in the physiological mechanism of diapause induction and not just for the expression of the diapause response. The light sensitivity threshold for photoperiodic induction of diapause was found to be extremely low, viz. less than 0.02 W/cm². Moreover, the light sensitivity threshold appeared to be strongly temperature dependent in A. potentillae. Experiments in which the mites experienced various sequences of short-day photoperiods and short-day thermoperiods, applied either concurrently or in succession, showed that the information collected by the photoperiodic counter and the thermoperiodic counter is integrated into one induction sum. These results strongly suggest that photoperiodic and thermoperiodic induction of diapause in these mites is based on the same physiological mechanism.Abbreviations DD continuous darkness - LL continuous light - LD light-dark cycle (e.g. LD 16:8 is a cycle of 16 h of light and 8 h of darkness) - TC thermoperiodic cycle (e.g. TC 16:8 (27°: 15°) is a thermoperiod with a 16 h thermophase of 27 °C and an 18 h cryophase of 15°C)     

Geometry of the dry-state oligomerization of 2′,3′-cyclic phosphates

Summary Evaporation of a solution of thymidine plus either theexo or theendo diastereomer of uridine cyclic 2,3-O, O-phosphorothioate (U > p(S) in 1,2-diaminoethane hydrochloride buffer gave the 2,5 and 3,5 isomers of (P-thio) uridylylthymidine (Up(S)dT) in a ratio of 1:2 with a combined yield of about 20%. These isomers were re-converted to U > p(S) and dT by a reaction that is known to proceed by an in-line mechanism. Both the 2,5 and 3,5 isomers gave as product the same diastereomer of U > p(S) that had been used originally in their formation. These dry-state prebiotic reactions (Verlander, Lohrmann, and Orgel 1973) are thus shown to be stereospecific, and both the 2,5 and 3,5 internucleotide bonds are formed by an in-line mechanism.Abbreviations DAE 1,2-diaminoethane - HPLC high pressure liquid chromatography - RNase bovine pancreatic ribonuclease A, EC 3.1.4.22 - TEAB triethylammonium bicarbonate - tris tris(hydroxymethyl)aminomethane - UMP(S) uridine monophosphorothioate - U > p uridine cyclic 2,3-phosphate - U > p(S) uridine cyclic 2,3-O, O-phosphorothioate - Up(S)dT (P-thio)uridylylthymidine - U2p(R_p-S)5dT (P-thio)uridylylthymidine with theR configuration at phosphorous, and a 2,5 internucleotide linkage     

Facultative parasites and host respiration II. Diseased tissue from which the pathogen has been removed

Summary Aseptic tissue cultures of Crown-gall ofScorzonera hispanica L. andHelianthus tuberosus L. have a higher rate of respiration than tissue cultures of the respective healthy tissue. Cultures of healthy tissue ofS. hispanica which have been subjected to high concentrations of auxin (napthalene acetic acid) until they reached an habituated stage in which they could grow without addition of the auxin, show an increased respiration similar to that of Crown-gall tissue cultures. It is concluded that the respiratory increase shown by Crown-gall tissue is not due to the retentive effect of a respiratory toxin alone but due also to a pathogen induced auxin metabolism.This work formed part of a Ph. D. thesis submitted to the Univ. of London, July 1961.     

Phytochrome dependent decrease of gibberellin-sensitivity

Exogenous gibberellin removes the genetical suppression of mesocotyl elongation in dark-grown seedlings of the rice cultivar Nihon Masari (japonica type). This gibberellin effect can be cancelled by light. All light effects can be accounted for by phytochrome. Dose-response and fluence-response studies show that phytochrome induces a reduction of the sensitivity to exogenous gibberellins. A cytological analysis of cell elongation and cortical microtubules led to a model where gibberellin and red light regulate mesocotyl elongation by controlling microtubule orientation in the epidermis of the mesocotyl. This causes corresponding changes of cellular extension growth, which can account for a large part of the observed growth responses. Comparative studies involving antimicrotubular drugs and gibberellin-synthesis inhibitors in the rice cultivar Kasarath (indica type) and a hybrid cultivar suggest that some of the differences between the cultivars are due to differences in gibberellin-sensitivity.     

Correlation between decreased reparative capability of UV lesions and higher spontaneous mutability in a mutator strain of E. coli K-12

Summary The biauxotrophic strain of E. coli K-12 (), met 1/his 7, which exhibits an 8 times higher rate of met1met ⁺-backmutation as compared with the parent strain met 1, was found to be also more sensitive to UV irradiation. In addition, the maximum UV induction of prophage occurs at lower doses, and the capacity of the strain to propagate induced prophage is reduced. The mutant strain has also lost part of the ability to repair UV-induced lesions in phage T 1.The phenotypes high mutability and UV sensitivity could not be separated by means of recombination.From these findings it is concluded that part of the spontaneously occuring changes in DNA (premutations) which lead to met ⁺ genotype are similarly repaired as some UV induced ones, and that the mutator mum ⁺ of strain met 1/his 7 causes a reduced repair of both changes.     

Extracellular α-amylase from Streptomyces rimosus

Summary A purification procedure for an extracellular -amylase from Streptomyces rimosus, oxytetracycline-producing strain, is described. The enzyme obtained was shown to be an acidic (pI 4.75) monomer with a relative molecular mass (M_r) of 43 000, containing three cysteines involved in the catalytic activity of the enzyme. Its amino-terminal part has 57–67% homology with amylases from other Streptomyces species. S. rimosus -amylase is sensitive to higher temperatures, and partially stabilized by Ca²⁺ ions. It hydrolyses starch (optimum at pH 5.0–6.0) in an endohydrolase manner giving rise to maltotriose, maltotetraose and higher oligosaccharides. Starch granules, except those from rice, were not significantly affected by the isolated -amylase.     

Gustatory intensity discrimination in rat NTS: a tool for the evaluation of neural coding theories

Summary The neural coding of taste quality in vertebrates currently is addressed by the labeledline and the across-fiber pattern theory. Experimental tests that have tried to distinguish between these theories by manipulating taste quality have been problematic in that both are fairly successful. However, the two theories maintain that different numbers of neurons participate in coding the presence of a substance. In the labeled-line theory, only those neurons labeled by that substance are supposed to be involved, whereas in the across-fiber pattern theory, all neurons responding to the substance are taken into account. We therefore expected that the theories might predict different intensity discriminating abilities among the four classical taste stimuli, which could provide a less equivocal test between the theories.A previous paper (Maes 1984) described a model for the neural basis of intensity discrimination, based on signal-detection theory. In the present paper, experimental data obtained from single neurons in the nucleus of the solitary tract of rat are analyzed in terms of this model. Whole-mouth stimulation with the four classical taste substances was used. For both theories, predictions of the intensity discriminating acuity for the four substances were derived, using different response analysis intervals, and including two plausible non-linearities in neural processing.The acuity profile across the four stimuli (consisting of the reciprocals of the four differential thresholds) turned out to be sensitive to the choice of coding theory and non-linearity. The interval chosen for response analysis had a strong effect. The predicted profiles should be checked against behavioral data on intensity discrimination in rat, but since such data are not yet available, a comparison was made with human psychophysical data, for illustrative purposes. It appeared that a unique combination of variables (tonic part of the response, across-fiber pattern coding with a contrast enhancement type of non-linearity) could be selected to match the psychophysical profile.In the Discussion, special attention is paid to the fundamental differences between the two coding theories, to the relevance of whole-mouth stimulation, and to the informational significance of various parts of the phasic/tonic response.Abbreviations AFP across-fiber pattern - CT chorda tympani - DA discrimination acuity - DT differential threshold - H sour - LL labeled line - N salty - NTS nucleus tractus solitarius - PTA pontine taste area - Q bitter - S sweet     

rho Mutations restore lamB expression in E. coli K12 strains with an inactive malB region

Summary lamB, the structural gene for receptor, is the second gene of the malK-lamB operon in the malB region of the Escherichia coli K12 chromosome. lamB is essentially not expressed in the absence of an active malT gene product, the activator or the maltose regulon. A malT strain is resistant to phage . We show that: (i) Introduction of rho mutations in malT mutants restores lamB expression to a level sufficient to render the strain sensitive to phage ; (ii) This restoration is not dependent on the main promoter of the malK lamB operon. It depends on the distal part of the malK gene.We propose that rho inactivation unmasks the activity of a promoter located near the distal end of malK. Experiments with Mu insertions in gene malK suggest that in the (-) orientation a Mu promoter is also able to allow lamB expression in a rho background.     

Direct and indirect effects of human interferon α on renal cell carcinoma: a new in vitro assay system for evaluating cytokine-mediated antitumor effects

A highly purified natural -interferon (nIFN) was tested in vitro for direct and indirect antiproliferative activity against renal cell carcinoma (RCC), using a modified human tumor clonogenic assay and clinically achievable concentrations. In preclinical experiments, the indirect (cytokine-mediated) antiproliferative activity of nIFN was investigated using ACHN cells (established human RCC cell line). Continuous exposure to nIFN at concentrations of more than 5 IU/ml in the presence of feeder cells (a mixture of 5×10⁴ monocytes/dish and 5×10⁵ lymphocytes/dish, obtained from healthy donors) significantly inhibited colony formation of ACHN cells in comparison with growth inhibition in the absence of feeder cells (P<0.05). Various cytokines were measured in the supernatants lying over the medium on the feeder-layer agarose containing the same conditioned feeder cells. With IFN at 500 IU/ml, tumor necrosis factor (TNF) and IFN were detected at markedly high levels for 2–24 h. Neutralizing anti-TNF monoclonal antibody significantly reduced the indirect antiproliferative activity. Using our modified human tumor clonogenic assay technique, sufficient numbers of colonies for drug testing were observed in 19 of 31 surgical specimens (61.3%). In these clinical materials, nIFN at a clinically achievable concentration (50 IU/ml) significantly inhibited colony growth in the presence of feeder cells consisting of 5×10⁴ monocytes/dish and 5×10⁵ lymphocytes/dish, obtained from the patient whose tumor was examined (P<0.05). In colony-forming cases, a significant correlation between the percentage colony survival and TNF concentration in the supernatant was observed (r=–0.95,P<0.01). These results suggest that this assay system may be an appropriate technique for evaluating the antiproliferative activities of nIFN involving cytokine-mediated action, and that TNF may play an important role in this cytokine-mediated activity.This work was supported in part by a grant-in-aid for promoting research (no. 02771010) from the Ministry of Education     

Carotenoid pigments of Sorangium compositum (Myxobacterales) including two new carotenoid glucoside esters and two new carotenoid rhamnosides

Summary The carotenoid pigments of the myxobacterium Sorangium compositum were analyzed by chromatographical and chemical techniques and by visible, infra red, and mass spectroscopy. Besides -carotene, neurosporene, torulene, lycopene, and 1,2-dihydro-1-hydroxy--carotene, four new carotenoid glycosides were found. These pigments were identified as 1,2-dihydro-1-hydroxy-torulene glucoside ester (I), 1,2-dihydro-3,1-dihydroxy-torulene glucoside ester (III), 1,2-dihydro-1-hydroxy-torulene rhamnoside (II), and 1,2-dihydro-3,1-dihydroxytorulene rhamnoside (IV).Fifth communication on the carotenoids of myxobacteria. Fourth communication see Arch. Mikrobiol. 76, 364–380 (1971).     

A monoclonal antibody chromosome marker analysis used to locate a loose smut resistance gene in wheat chromosome 6A

Many genes have been located in wheat chromosomes, yet little is known about the location of genes for resistance to Ustilago tritici, which causes loose smut. Crosses were made between the loose smut susceptible alien substitution lines Cadet 6Ag(6A) and Rescue 6Ag(6A) (lines in which Agropyron chromosome 6 is substituted by wheat chromosome 6A) and four cultivars resistant to U. tritici race T19: Cadet, Kota, Thatcher and TD18. The segregating progeny were tested for reaction to race T19 and for the level of binding with a monoclonal antibody specific to a chromosome 6A-coded seed protein. The antibody, which does not bind to seed protein extracts in the absence of the 6A chromosome, was used as a chromosome marker. An association was established between resistance to race T19 and the presence of chromosome 6A for each of the cultivars tested, indicating that resistance to race T19 resides in chromosome 6A. Ustilago tritici race T19 resistance in Cadet appears to be located in the short arm of chromosome 6A, based on the evaluation of the Cadet 6A long ditelosomic stock, which was susceptible, and the Cadet 6A-short: 6-Agropyron- short alien translocation stock, which was resistant.     

The comparative effects of IL-1 and TNF on AMN-anti-Ly 2.1 immunoconjugate therapy

The administration of mTNF and hIL-1 was investigated for their potential to increase the anti-tumor activity of AMN-anti-Ly-2.1 against the Ly-2.1⁺ murine thymoma ITT(1) 75 NS E3. Dose response studies using mTNF alone demonstrated a single 10g iv injection produced 30% inhibition in tumor size while 3 doses of 1g administered on alternative days produced 70% tumor inhibition. By contrast, hIL-1 was unable to significantly reduce E3 tumor size using single doses up to 10g or a total of 30g administered in 3 doses (iv or ip). However, intratumor injection of hIL-1 (20g injected in 2 doses) produced 20% inhibition in tumor size. Combination therapy using AMN immunoconjugates with mTNF showed enhanced antitumor activity compared to each agent alone. Biodistribution studies revealed that anti-tumor activity, was due to increased localization (2–3 fold) of AMN immunoconjugates in the presence of mTNF- whereas huIL-1 was without effect unless accompanying toxicity was seen. Clearly for this tumor, mTNF potentiated the effects of AMN immunoconjugates. Despite the shared biological properties of these cytokines, mTNF is superior to hIL- for augmenting drug immunoconjugate.Abbreviations AMN Aminopterin - CBF₁ (C57BL6xBALB/c)F₁ mice - DMSO Dimethyl sulfoxide - E3 ITT(1)75NS E3 - HAMA human-anti-mouse antibody - i.p. intraperitoneal(ly) - I.T. intratumor - i.v. intravenous(ly) - hIL-1 recombinant human Interleukin-1-alpha - MoAb monoclonal antibody - PBS phosphate buffered saline - SE standard error - s.c. subcutaneous(ly) - mTNF recombinant murine tumor necrosis factor-alpha     

Effect of alpha-isopropylmalate on the synthesis of RNA and protein in Neurospora

Summary The leu-3/-IPM (-isopropylmalate) regulatory system, previously shown to control several genes of leucine, isoleucine, valine, and histidine biosynthesis, appears likely to be involved also in the regulation of overall RNA and protein synthesis in Neurospora. Upon addition of -IPM the synthesis of all major species of stable RNA was found to be transiently inhibited by approximately 50%. A similar reduction was observed in overall protein synthesis. The inhibition was dependent in both cases on a functional leu-3 gene product, in conformance with previously established patterns of -IPM dependent gene regulation. The overt resemblance of the phenomenon described here to the stringent response of bacteria is noted but neither the mechanism of inhibition nor the precise role of -IPM in the process has been established.