Biosynthesis of RNA polymerase in Escherichia coli

Summary Effect of temperature-sensitive, assembly-defective mutations in Escherichia coli RNA polymerase (rpoB) or subunit gene (rpoC) was investigated on the expression of wild-type rpoB ⁺C⁺operon, which was introduced by infection of a lambda transducing phage drif ⁺ (rpoB ⁺)-6 after UV-irradiation of the mutant cells. In rpoB2·rpoB7 strain which accumulates assembly-intermediates, free , ₂ complex and premature core, the expression of rpoB ⁺C⁺operon measured by the rate of subunit synthesis was considerably inhibited whereas that of EF(translation elongation factor)-Tu, ribosomal proteins L1 and L7/L12, and some -coded proteins remained unaffected. On the other hand, the expression was enhanced specifically for only rpoB ⁺C⁺operon in either rpoC4 or rpoC1 mutants, which are defective in the association of ₂ complex and subunit or the activation of premature core enzyme, respectively. Upon preincubation of the mutant cells at 42° C prior to phage infection, during which assembly intermediates degraded rapidly, the rate of subunit synthesis relative to other phage-corded proteins increased remarkably in rpoB2·rpoB7 mutant as well as in rpoC4 and rpoC1 mutants. These observations strongly suggested the autogenous regulation for at least (rpoB ⁺C⁺) operon by some trans-active diffusible protein complexes built of RNA polymerase subunits. Nature of the regulatory molecules is discussed.Paper VI in this series is Saitoh and Ishihama (1977)     

Idioms of distress: Kinship and sickness among the people of the Kingdom of Tonga

Idioms of distress refers to the popular expression of emotional tension that arises in the relationship between sickness and kinship. By reference to case studies and discussions among the Polynesian people of Tonga, the author shows where such tension arises and how it influences the sickness process. Sickness is necessarily a collective phenomenon which can best be understood not simply as a clinical event, but as an experience that is part of the experience of family. Various ways of expressing distress as a reflexive encounter between personal and cultural meaning systems are reviewed, as are several new concepts such as doing sickness as kinship, and turning in the process of decision making in the kinship management of sickness. The explanatory models of sickness in Tonga are shown to encompass culturally sanctioned expressions of distress as part of the adaptive coping mechanisms in that society. Distress frequently emerges in somatic form, as a number of studies have shown. However, the author emphasizes the kinship meaning of sickness, kinship management and sickness therapy, the adaptive process of idiomatic expressions of distress, which are expanded here and offered as potential avenues for elaboration in other cultural milieu. Two aspects of the notion idioms of distress are noted, and the phenomenon is understood as a process which acts as a prime mover in social change.     

Nucellar callus of ‘Femminello’ lemon,selected for tolerance toPhoma tracheiphila toxin,shows enhanced release of chitinase and glucanase into the culture medium

Phoma tracheiphila is the causative agent of the disease mal secco. Citrus cultivars differ substantially in respect to their sensitivity to the pathogenP. tracheiphila and its toxin. Some cultivars (e.g., Femminello lemon) are inherently sensitive while others (e.g., Tarocco orange) are tolerant. Cell lines derived from nucellar tissue of Femminello, Tarocco and a cell line selected for tolerance to the fungal toxin (Femminello-S) were used to study host-pathogen interaction. Our results showed that calli or conditioned media of Tarocco and Femminello-S inhibited the size of co-cultivated fungal colonies when compared to Femminello. In addition, conditioned medium of Tarocco as well as FemminelloS, but not Femminello, promoted bursting of hyphal tips. A ten-fold increase in chitinase and glucanase enzymatic activity, as evaluated by radiometric assay and laminarin hydrolysis respectively, was detected in Femminello-S extracellular extracts as compared to Femminello. An increase in chitinase was also shown by immunoblot analysis. Our findings suggest a positive correlation between the presence of chitinase and glucanase in the conditioned media of the cultured cells and the tolerance of those cells toP. tracheiphila toxin.     

Optomotorische Untersuchung des visuellen systems einiger Augenmutanten der Fruchtfliege Drosophila

Summary The optical properties of the compound eye of Drosophila have been analysed using the optomotor reactions of flies with normal and mutant eye pigmentation. The stimulus was provided by cylindrical patterns with different periodic intensity distributions rotating at different speeds. The response consists of a torque about the vertical axis and was recorded under conditions of fixed flight. (Maximum reaction is about 0.04 dyn · cm). The transfer characteristics of the optical system are determined by the inter-ommatidial angle , influencing the resolving power and by the width of the visual field of single ommatidia , influencing the response at high spatial frequencies. The values = 4.6° and = 3.5° are obtained from stimulus-response experiments with Drosophila. They are independent of the presence of screening pigments. Differences in the response of flies with strong (+, se), weak (w ^a), and missing (w) pigmentation can be explained by the increased amount of scattered light in the pigment-deficient eyes. The overall intensities in the equally illuminated receptors are expected to be in the ratio 11825, respectively. The perception of motion depends only on the temporal, not on the spatial phase relations between periodic intensity variations in neighbouring ommatidia. Therefore the inhomogeneous distribution of the inter-ommatidial angle changes the resolving power in different parts of the eye without changing the response to motion. Different simultaneous stimuli of equal strength in different parts of the eye are averaged in the perceptive system of Drosophila according to the number of ommatidia in these parts.     

Comparative study on the chloroplast,mitochondrial and nuclear genome differentiation in two cultivated rice species,Oryza sativa and O. glaberrima,by RFLP analyses

Summary Restriction fragment length polymorphisms of chloroplast (ct), mitochondrial (mt) and nuclear DNA were investigated using eight cultivars of Oryza sativa and two cultivars of O. glaberrima. Relative variability in the nuclear and cytoplasmic genomes was estimated by a common measure, genetic distance. Based on the average genetic distances among ten cultivars for each genome, the evolutionary variabilities of the mitochondrial and nuclear genomes were found to be almost the same, whereas the variability of the chloroplast genome was less than half that of the other two genomes. Cluster analyses on ct and mt DNA variations revealed that chloroplast and mitochondrial genomes were conservative within a taxon and that their differentiations were well-paralleled with respect to each other. For nuclear DNA variation, an array of different degrees of differentiation was observed in O. sativa, in contrast with little variation in O. glaberrima. As a whole, differentiation between O. sativa and O. glaberrima was clearly observed in all three genomes. In O. sativa, no notable difference was found between the cultivars Japonica and Javanica, whereas a large differentiation was noticed between Japonica (including Javanica) and Indica. In all three genomes, the average genetic distances within Indica were much larger than those within Japonica (including Javanica), and almost similar between Japonica (including Javanica) and Indica. These facts indicate that differentiation in O. sativa was due mainly to Indica.     

High-level Expression of Maize <Emphasis Type="Italic">γ</Emphasis>-zein Protein in Transgenic Soybean (<Emphasis Type="Italic">Glycine max</Emphasis>)

Primers were developed for 118 microsatellites isolated from grape (Vitis vinifera) genomic libraries enriched for (AC)n repeats. Only one microsatellite sequence matched other grape SSR-sequences in the GeneBank database. Genotyping was carried out in the parental lines and four offspring of two pseudo-test-cross populations, Cabernet Sauvignon x Seyval and Chardonnay x Bianca, and a further six other grape genotypes (V. vinifera Sultanina, Merlot, Syrah, Müller-Thurgau, Vitis Regent and V. riparia Gloire de Montpellier). A total of 108 microsatellites showed easily scorable alleles and 100 of them segregated according to a configuration suitable for mapping in either cross. A further 8 SSRs, although unsuitable for mapping in those crosses, showed polymorphism in the other genotypes tested. This set of markers was used, along with 75 microsatellites of other repeat-types, to fingerprint 46 offspring of the cross Chardonnay x Bianca. For each full-sib, individual heterozygosity and distance in repeat units between pairs of alleles at each locus (mean d²) were calculated as a tool for predicting highly outbred recombinant individuals. Six microsatellites with segregation ratios significantly distorted towards the lack of homozygous sibs were identified and mapped to linkage groups LG 3 and LG 5. Estimation of heterozygosity at genome-wide level and genotyping at loci for which homozygous sibs are discriminated against are discussed for marker-assisted background selection in outcrossing grapevines.     

ααααanti-3.7 type II: a new α-globin gene rearrangement suggesting that the α-globin gene duplication could be caused by intrachromosomal recombination

Summary We report here a new human -globin gene rearrangement carrying the two normal, 2 and 1, and two hybrid, 1/2, globin genes in the order 5-2-1/2-1/2-1-3. Both the hybrid genes, subtyped with ApaI and RsaI restriction enzymes, were found to be of the uncommon anti 3.7 type II. The hybrid genes were expressed at the biosynthetic level and their interaction with the -thalassaemia IVS 1 nt 1 GA mutation caused thalassaemia intermedia. We also report a case of an -globin gene rearrangement in the twin of one of the -globin gene carriers; the duplicated gene was of the anti 4.2 type and was associated with the absence of RsaI polymorphism. The singular finding of an -anti 3.7 cluster with two identical rare hybrid genes suggests that the reciprocal unequal recombination causing the -globin gene rearrangements could be of the intra-chromosomal rather than the interchromosomal type.     

Evaluation of structural and physiological plant characteristics in relation to the distribution of cadmium in maize inbred lines

To establish the structural and physiological characteristics related to the genotypic variation in Cd distribution between maize inbred lines (shoot Cd excluders and non-shoot Cd excluders), shoot and root morphological parameters were studied on plants grown in nutrient solution. Furthermore, the xylem sap composition and the desorbability of Cd from roots of these inbreds have been compared. No relationship between the morphological characteristics of either shoot (specific leaf area and leaf area ratio) or root (specific root length, specific surface area, and average diameter) and Cd distribution could be assessed. Cadmium concentrations in the xylem exudates from non-shoot Cd excluders were higher than those from shoot Cd excluders, but not related to citrate and malate concentrations. The absolute and relative amounts of Cd desorbed from roots of shoot Cd excluders were about twice as high compared to those of the non-shoot Cd excluders, especially at the lowest Cd concentration in solution. The absence of a relationship between shoot or root morphological parameters and Cd partitioning and the differences between both groups in the amounts of Cd desorbed, even at similar root Cd concentrations, indicate that the differential Cd distribution between shoot Cd excluders and non-shoot Cd excluders may be related to the observed differences in root Cd concentration, desorption characteristics and binding capacity of Cd inside and/or outside the root and its distribution within the roots.     

Structures of the N-linked sugar chains in the PAS-6 glycoprotein from the bovine milk fat globule membrane

The structures of the N-linked sugar chains in the PAS-6 glycoprotein (PAS-6) from the bovine milk fat globule membrane were determined. The sugar chains were liberated from PAS-6 by hydrazinolysis, and the pyridylaminated sugar chains were separated into a neutral (6N) and two acidic chains (6M and 6D), the acidic sugar chains then being converted to neutral sugar chains (6MN and 6DN). 6N was separated into two neutral fractions (6N13 and 6N5.5), while 6MN and 6DN each gave a single fraction (6MN13 and 6DN13). The structure of 6N5.5, which was the major sugar chain in PAS-6, is proposed to be Man16 (Man13) Man14GlcNAc14GlcNAc-PA; 6N13, 6MN13 and 6DN13 are proposed to be Gal13Gal14GlcNAc12Man16 (Gal13Gal14GlcNAc12Man13) Man14GlcNAc14 (Fuc16)GlcNAc-PA;6M and 6D had 1 or 2 additional NeuAc residues at the non-reducing ends of 6MN13 and 6DN13, respectively. © 1998 Rapid Science Ltd     

Synthesis of Gal-β-(1→4)-GlcNAc-β-(1→6)-[Gal-β- (1→3)]-GalNAc-α-OBn oligosaccharides bearing O-methyl or O-sulfo groups at C-3 of the Gal residue: specific acceptors for Gal: 3-O-sulfotransferases

Our recent studies have revealed the existence of two distinct Gal: 3-O-sulfotransferases capable of acting on the C-3 position of galactose in a Core 2 branched structure, e.g., Gal14GlcNAc16(Gal13)GalNac1OBenzyl as acceptor to give 3-O-sulfoGal14GlcNAc13(Gal13)GalNAc1OB 20 and Gal14GlcNAc16(3-O-sulfoGal13)GalNAc1OB 23. We herein report the synthesis of these two compounds and also that of other modified analogs that are highly specific acceptors for the two sulfotransferases. Appropriately protected 1-thio-glycosides 7, 8, and 10 were employed as glycosyl donors for the synthesis of our target compounds.     

An elongated segment of DNA observed between two human α globin genes

Summary Detailed restriction enzyme analysis of the DNA from a Chinese female showed that one of her chromosomes had a >17.5 kb deletion of DNA, including the , 2, and 1 globin genes, which is present in many Southeast Asians with an -thalassemia-1 chromosome. Her normal chromosome had the expected cluster of -like globin genes (5----2-1-3), but the segment of DNA between the two globin genes was elongated by some 0.5–0.7 kb. Analyses of various restriction sites suggested that this normal variant of the human globin gene complex is due to a crossover between a normal chromosome with () and a chromosome with an -thalassemia-2 (–^3.7) and an -21-hybrid gene.     

Ecological theories and Dutch nature conservation

This paper aims to achieve insight into various ecological theories in the Netherlands which have different, and sometimes opposing, views on the conservation of nature. Interviews, publications and archival research brought to light four separate theories: vitalistic/holistic, dynamic, cybernetic and chaos. Diversity is reached through stability according to vitalistic/holistic and cybernetic theories, but through change and instablility according to the dynamic and chaos theories. These two groups are working apart, and continue to have their own ideas. Prediction of the future is only possible with the vitalistic/holistic and cybernetic theories. Ecologists who adhere to these theories feel responsible and able in different ways to change ecological nature towards desirable end goals. The other two theories, dynamic and chaos, appear to be less activist.     

Biological,chemical and hydrological characters of coastal lagoons of Ghana,West Africa

Two types of lagoon which occur along the West Coast of Africa described as open and closed, were studied using oceanographic methods. The parameters were temperature, salinity, dissolved oxygen and productivity. It was observed that rainfall, evaporation and the tides have a combined effect on the variations in the temperature and the salinity. The opening of the lagoons into the sea affects the level of the productivity and the rate at which salinity increases with time.Minimum temperatures at the heads of the lagoons were found to be lower than those near the ocean, while the maximum temperatures were higher at the heads. During the dry period, due to continuous evaporation of water, the salinity in the closed lagoon usually rises steeply, while those in the open lagoon rise gently.The levels of dissolved oxygen concentration in the open lagoon are higher than those in the closed lagoon. This is the result of the fact that phytoplankton count in the open lagoon is higher than that in the closed lagoon. There is also a greater diversity of phytoplankton and more live phytoplankton cells per cubic centimetre of water in the open lagoon than in the closed lagoon.As a result of all this, the main organic productivity in the open lagoon is higher than those in the closed lagoon.     

Multiple shoot regeneration from root organ cultures of Populus alba x P. grandidentata

Excised roots of various ages from Crandon and Hansen clones of Populus alba x P. grandidentata were tested for their regeneration capacity. Sixty-day-old excised roots that contained root tips were found to be most suitable. The highest number of shoots (an average of 111 shoots/root segment with Crandon and 98 with Hansen) was obtained by adding 22M and 14M zeatin to the medium, respectively. The two clones of hybrid poplar responded similarly to growth regulator treatments; however, the number of shoots produced was greater from the root organs derived from Crandon clones. Regenerated shoots were rooted in basal Woody Plant Medium without any growth regulators. Successful transplantation into soil and growth was achieved with all plants.     

The effect of specific chromosome and cytoplasm substitutions on the tissue culture response of wheat (Triticum aestivum) callus

Summary Calli were initiated from immature embryos of four lines of hexaploid wheat (Triticum aestivum L. em. Thell), the euplasmic nuclear donor Chinese Spring, Chinese Spring in which both 4B chromosomes were substituted by those of the variety Cappelle-Desprez and two alloplasmic lines in which these nuclei were substituted into the cytoplasm of Aegilops ovata. The calli were found to differ in their initia growth rates and their ability to organise shoot primordia and regenerate shoots. The Cappelle 4B chromosomes had a very significant effect on all these characters. The potential for modelling genotypes for improved tissue culture characteristics is discussed.     

Plant regeneration from Citrus protoplasts: Variability in methodological requirements among cultivars and species

Summary Nucellar callus lines were established from two orange cultivars (Nucellar Shamouti, Shamouti Landau), three mandarin cultivars (Murcott, Dancy, Ponkan) one grapefruit cultivar (Duncan) and sour orange (Citrus aurantium). These callus lines were initiated from in vitro cultured ovules of young fruits and maintained an embryogenic capacity. The plating efficiencies of protoplasts derived from these calli, as well as those of protoplasts from lemon (cv. Villafranca) nucellar callus were differentially affected by the maceration enzymes and by the sugars used as osmotic stabilizers. Plants with normal morphological features were regenerated from cultured protoplasts derived from each of the nucellar callus lines. The establishment of eight new protoplast systems in Citrus paves the way for cell genetics studies and for novel breeding approaches in these economically important orchard trees.Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. E-290, 1981 series     

Poly(U)-directed peptide-bond formation from the 2′(3′)-glycyl esters of adenosine derivatives

Summary The self-condensation of 2(3)-O-glycyl esters of adenosine, adenosine-5-(O-methylphosphate) and P₁, P₂-diadenosine-5-pyrophosphate in 6.2 mM solutions at pH 8.0 and -5°C in the presence of 12.5 mM poly(U) yields approximately 3 times as much diketopiperazine as reactions without poly(U). As the concentration of 2(3)-O-(glycyl)-P₁, P₂-diadenosine-5-pyrophosphate is decreased from 6.2 mM to 1.5 mM the yield of diketopiperazine in the presence of poly(U) decreases slightly from 6.6% to 5.2%, whereas, in the absence of poly(U) the yield of diketopiperazine decreases substantially from 2.4% to 0.75%. The enhanced yield of diketopiperazine that is attributed to the template action of poly(U) is temperature dependent and is observed only at temperatures below 10°C (5°C to -5°C) for 6.2 mM 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) and below 23°C (15°C to -5°C) for 6.2 mM 2(3)-O-(glycyl)-P₁, P₂-diadenosine-5-pyrophosphate. The absence of a template effect at high temperatures is attributed to the melting of the organized helices. The hydrolysis half-lives at pH 8.0 and -5°C of 2(3)-O-(glycyl)-adenosine, 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate), 2(3)-O-(glycyl)-P₁, P₂-diadenosine-5-pyrophosphate, and 5-O-(glycyl)-adenosine in the presence of poly(U) are substantially larger than their half-lives in the absence of poly(U). The condensation of 2(3)-O-(glycyl)-adenosine yields 5% of 5-O-(glycyl)-adenosine in the presence of poly(U) compared to 0.7% in the absence of poly(U).Abbreviations DKP diketopiperazine - (gly)₂ glycylglycine - (gly)₃ glycylglycylglycine - AppA-gly 2(3)-O-(glycyl)-P₁, P₂-diadenosine-5-pyrophosphate - MepA-gly 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) - Ado-2(3)-gly 2(3)-O-(glycyl)-adenosine - Ado-5-gly 5-O-(glycyl)-adenosine - Boc-gly N-tert-butyloxycarbonylglycine - AppA P₁, P₂-diadenosine-5-pyrophosphate - MepA adenosine-5-(O-methylphosphate) - AppA-Boc-gly 2(3)-O-(Boc-glycyl)-P₁, P₂-diadenosine-5-pyrophosphate - Ado-5-Boc-gly 5-O-(Boc-glycyl)-adenosine - Ado-2(3)-Boc-gly 2(3)-O-(Boc-glycyl)-adenosine     

Screening of turfgrass species and cultivars for NaCl tolerance

Summary Information regarding the relative levels of salt tolerance between cultivars of Kentucky bluegrass (Poa pratensis L.) is lacking. The objectives of this study were to 1) develop a simple, quick and sensitive method of screening turfgrass species for NaCl tolerance and 2) to compare the relative salt tolerance of five cultivars of Kentucky bluegrass (Ram I, Adelphi, Baron, Bensun, and Nassau) to other known salt tolerant turfgrass species such as alkalaigrass (Puccinellia distans (L.) Parl. cv. Fults) and two cultivars of red fescue (Festuca rubra L. Dawson, and Checker).Alkalaigrass and both cultivars of red fescue retained a high level of salt tolerance compared to the Kentucky bluegrass cultivars. Significant variability in salt tolerance was apparent among the Kentucky bluegrass cultivars with Adelphi and Ram I exhibiting the best overall tolerance.     

Patterns of direct and indirect embryogenesis from mesophyll protoplasts of Medicago sativa

Clones of three cultivars of Medicago sativa (Rambler, Regen S and Rangelander) were used as sources of mesophyll protoplasts. Although all three clones readily produced protoplasts, the subsequent development patterns in culture varied greatly among genotypes, with protoplasts from Regen S and Rambler forming calli which could be induced to form embryos, and protoplasts from Rangelander undergoing direct embryogenesis. Protoplasts of Regen S exhibited high rates of division while those of Rangelander tended to aggregate with only a few cells per aggregate surviving. The surviving cells gave rise to proembryos within the aggregates; these proembryos developed into differentiated embryos after 5–7 weeks of culture. Based on the initial protoplast population, the efficiency of embryo formation averaged 0.13% and ranged from 0.001–0.4%. Observations during the early stages of culture indicated that cell aggregation was a prerequisite for direct embryogenesis.     

Cytogenetic analysis of structural rearrangements in three varieties of common wheat,Triticum aestivum

Summary The winter wheat varieties Starke and Cappelle Desprez and the spring wheat Chinese Spring were analysed for structural chromosome rearrangements that resulted in the formation of multivalents in F₁ hybrids. The analyses were carried out using hybrids involving euploids, monosomic and ditelosomic stocks, and double-monotelodisomic constructs. The study confirmed that Cappelle Desprez differs from Chinese Spring in a reciprocal translocation between chromosomes 5B and 7B (Riley et al. 1967); a translocation involving chromosomes 3B and 3D could not be verified. Furthermore, the analysis showed that Starke differs from Chinese Spring in a reciprocal translocation between chromosomes 7A and 7D. Both translocations have a coefficient of multivalent realisation of about 0.84. Further multivalents in euploid Starke, in euploid and some aneuploid stocks of Cappelle Desprez, and in euploid as well as various types of aneuploid hybrids between all three varieties could nearly all be explained hypothesizing that chromosome 2B of both Starke and Cappelle Desprez is a duplication-deficiency chromosome. In the hypothesis a part of the long arm of 2B is missing and replaced by a duplicated part of the long arm of chromosome 2D. The multivalents of this rearrangement showed an average coefficient of realisation of about 0.09.Sven Ellerström died in December 1985