The effect of temperature and genetic background on the phenotypic expression of severalVestigial strains ofDrosophila melanogaster

The eightvestigial strains,vg, vg-co-iso,vg;se-co-iso,vg-ms-1-co-iso,vg-ms-1; 3-co-iso,vg-ms-co-iso,vg-ms;se andvg-ms;se-co-iso showed various temperature responses. The five strains of thevg-ms group showed a greater response to temperature than did the three strains of thevg group. This tendency became more pronounced the higher the temperature was. The difference in temperature response between thevg-ms-1; 3-co-iso andvg-ms-co-iso strains indicates that the phenotypic expression ofvestigial is influenced by a modifier or modifiers located on the second chromosome of the Oregon(iso) strain. It was found that the X chromosome of the Oregon(iso) strain showed a slight modifier action in females but not in males of thevg-ms mutant.These gene in thevg-ms;se strain seemed to enhance the sensitivity to heat, and to inhibit the emergence of adult flies from pupal cases at 30°C when combined with thevg-ms gene, but no interaction was seen between these gene and thevg gene. From the results of this experiment, it is assumed that thevg-ms mutant either has a new recessive allele of thevg gene, or a modifier gene(s) closely linked tovg.This work forms part of a thesis for the doctorate of Kyoto University.     

Reversal effect of the genetic background on some small-eye mutants,with special reference to gene action of morphogenetic mutants inDrosophila

The facet number of heterozygous females of theBar-eye mutant ofDrosophila melanogaster varies from ±154 to ±665 according to the full eye strains combined with the mutant. TheBar character is commonly recognized as a semidominant, but its manifestation varies from nearly complete dominance to recessiveness according to the mode of the heterozygous combination. Virtually the same is true of the mutantLobe ² which is usually acknowledged as a complete dominant, but may be manifested as an incompletely dominant gene. Thew ^e andb gp strains decrease the eye size of the heterozygotes withBar, but increase the eye size of the heterozygotes withLobe ².Co-isogenic strains ofBar, bar-3 andeyeless ² mutants with Oregonisogenic increase the facet number to degrees 1.4–1.8 times of the original strains. However, the co-isogenicLobe ² strain with the same Oregonisogenic strain reduces the facets completely, and makes virtually all individuals eye-less. These observations indicate that the same genetic background may give an opposite effect to different small-eye mutants. Analysis was carried out for modifying genes ofbar-3. Modifiers were detected on all large chromosomes, besides a recessive major gene located on the 2nd chromosome of Oregon-isogenic strain.A part of this work was performed in the Division of Biology, California Institute of Technology, Pasadena.     

The Effect of the Genotype on the Expressiveness of Character vestigial and Polyteny of Giant Chromosomes in Drosophila melanogaster Meig.

The effects of the Bar (B) andwhite (w) mutations on the expressiveness of the character vestigial (vg) and the degree of polyteny of salivary gland giant chromosomes were studied in Drosophila melanogaster.Either mutation changed both the expressiveness of vestigial and the degree of chromosome polyteny. A negative association between the vg expressiveness and the degree of chromosome polyteny was revealed and proved to be stronger in females than in males. The parameters under study were shown to differ between females and males.     

Specification of Primary Pigment Cell and Outer Photoreceptor Fates byBarH1Homeobox Gene in the DevelopingDrosophilaEye

In the developingDrosophilaeye,BarH1andBarH2, paired homeobox genes expressed in R1/R6 outer photoreceptors and primary pigment cells, are essential for normal eye morphogenesis. Here, we show evidence thatBarH1ectopically expressed under the control of thesevenlessenhancer (sev-BarH1) causes two types of cone cell transformation: transformation of anterior/posterior cone cells into outer photoreceptors and transformation of equatorial/polar cone cells into primary pigment cells.sev-BarH1repressed the endogenous expression of theroughhomeobox gene in R3/R4 photoreceptors, while theBarH2homeobox gene was activated bysev-BarH1in an appreciable fraction of extra outer photoreceptors. In primary pigment cells generated by cone cell transformation, the expression ofcut,a homeobox gene specific to cone cells, was completely replaced with that ofBarhomeobox genes. Extra outer photoreceptor formation was suppressed and enhanced, respectively, by reducing the activity of Ras/MAPK signaling and by dosage reduction ofyan,a negative regulator of the pathway, suggesting interactions betweenBarhomeobox genes (cell fate determinants) and Ras/MAPK signaling in eye development.     

Vitamin B12 transport in Escherichia coli K12 does not require the btuE gene of the btuCED operon

Summary Transport of vitamin B₁₂ across the cytoplamic membrane ofEscherichia coli requires the products ofbtuC andbtuD, two genes in thebtuCED operon. The role ofbtuE, the central gene of this operon, was examined. Deletions withinbtuE were constructed by removal of internal restriction fragments and were crossed onto the chromosome by allelic replacement. In-frame deletions that removed 20% or 82% of thebtuE coding region did not affect expression of the distalbtuD gene. These nonpolar deletions had little effect on vitamin B₁₂ binding (whole cells or periplasmic fraction) and transport. They did not affect the utilization of vitamin B₁₂ or other cobalamins for methionine biosynthesis, even in strains with decreased outer membrane transport of vitamin B₁₂. ThebtuE mutations did not impair adenosyl-cobalamin dependent catabolism of ethanolamine or repression ofbtuB expression. Thus, despite its genetic location in the transport operon, thebtuE product plays no essential role in vitamin B₁₂ transport.     

Mechanism of induction ofBar-like eye malformation by transient overexpression ofBar homeobox genes inDrosophila melanogaster

TheBar locus ofDrosophila is known to be a small complex consisting of two similar homeobox genes,BarH1 andBarH2. Usingegr as an ommatidium marker, possible mechanisms of formation of malformed eyes were examined. As in the case ofBarH1, overexpression ofBarH2 was found to be capable of inducingBar-like eye malformation. It was suggested that suppression of the anterior progression of the morphogenetic furrow and inhibition of reinitiation of normal ommatidial differentiation were mandatory to formation of the reduced eye morphology inBar mutants. These authors equally contributed to the present paper.     

Cell degeneration and elimination in the imaginal wing disc,caused by the mutationsvestigial andUltravestigial ofDrosophila melanogaster

Summary The mutationsvestigial (vg; recessive) andUltravestigial (vg ^U; dominant) ofDrosophila melanogaster give rise to identical mutant adult phenotypes in which much of the cases this results from cell death in the presumptive wing margin of the wing disc in the third larval instar, but the process of cell degeneration is quite different in the two mutants. Invg cell death occurs continuously throughout the third larval instar, while invg ^U it occurs only in the early third instar. Cells fragment and some of the fragments condense, becoming electron dense (apoptosis). Both condensed and ultrastructurally normal cell fragments are extruded to the basal side of thevg disc epithelium. They accumulate under the basal lamina in the wing pouch area until they are phagocytosed by blood cells entering the wing pouch during the six hours following pupariation. Fragments are not extruded from thevg ^U epithelium but are apparently phagocytosed by neighboring epithelial cells. The basal lamina undergoes mophological changes following pupariation and is phagocytosed by blood cells in both wild-type andvestigial, but investigial the degenerated cell fragments are also engulfed by the same blood cells.     

New palynological and tephrostratigraphical investigations of two salt lagoons on the island of Mljet, south Dalmatia, Croatia

In the sediments of both of the investigated lakes, the tephra from the Mercato-Ottaviano eruption (Vesuvius, southern Italy) (ca. 7900 B.P.) could be identified. The palynological investigations show that from ca. 9000-7200 B.P. (8000-6000 cal B.C.) deciduous oak forests predominated, with only a few representatives of Mediterranean vegetation. At the transition to the central European Atlantic Period those forests changed to an open vegetation type, dominated byJuniperus andPhillyrea. At about 5500 B.P. (4400 cal B.C.), theJuniperus-Phillyrea vegetation was replaced byQuercus ilex woodland that still occurs on the island of Mljet today and is considered to be the natural vegetation of the Dalmatian coastland. The associated vegetation of theQ. ilex forests changed several times. At the beginning of theQ. ilex period,Juniperus values were still high, but soon they decreased andErica spread. In more recent times theQ. ilex forests were partially replaced by plantations ofPinus halcpensis. Indicators of human impact are sparse throughout the pollen record. Clear evidence for human influence exists only from ca. 3100 B.P. (1300 cal B.C.) whenJuglans andPinus halepensis were introduced to the area. Later,Olea andSecale cultivation can be suggested and further spreading ofJuniperus indicates use of the land as pasture.     

Regulation of biosynthesis of pesticidal metabolic complexes inStreptomyces griseus

The complex of pesticidal metabolites produced byStreptomyces griseus LKS-1 consists of a peptide antibiotic (A), nonactic acids (B), macrotetrolides (C), pyrrolizines (D), and of cycloheximide. The latter unwanted phytotoxic compound was eliminated by treatment with mutagens. Combined approaches, including both genetic and physiological manipulations, resulted in the following alterations in the biosynthetic capacity: (1) A more than 80-fold increase in the production of C under a substantial decrease in the yields ofA, B andD, the ratio of the components ofC being steered toward the required more active ones; (2) a more than 300-fold increase in the production ofB under suppression of the formation ofA andC: (3) a 10-fold increase in the yields ofD under suppression ofA andC; (4 a significant increase in the yields ofA with eliminatingB, C andD. The level of inorganic phosphate in fermentation media and the sensitivity of the organism to carbon catabolite repression were important factors participating in the regulation of the above biosynthetic processes.     

Relationships in theAcanthaceae and related families as suggested by cladistic analysis ofrbcL nucleotide sequences

A parsimony analysis of DNA sequences of the chloroplast-encoded generbcL from twelve members of theAcanthaceae s.l., including members of the sometimes segregateThunbergioideae andNelsonioideae, and other families in theBignoniales sensuThorne (1992) is presented. The results largely agree with the classification of theAcanthaceae presented byBremekamp (1965) andThorne (1992) and supportNelsonioideae as a sister group to the rest of theAcanthaceae. Thunbergioideae are placed as a sister toAcanthaceae s.str.Acanthus andAphelandra, both representatives ofAcanthoideae, form a sister group toRuellioideae. An analysis of branch support found that many branches throughout theBignoniales are weakly upheld. This points to the need for further studies in the group using more sequences ofrbcL as well as other data. None of the families ofBignoniales as presently circumscribed (includingAcanthaceae s.l.) were strongly supported, although the larger clade containing the families of theBignoniales was robust.     

Genetic analysis of rec E activities in Bacillus subtilis

Summary ArecE mutant (recE6) ofBacillus subtilis was constructed by insertion of a selectable marker into therecE coding region. The insertional inactivation of therecE gene renders cells very sensitive to DNA damaging agents and severely impairs intermolecular recombination, but does not markedly affect plasmid interstrand annealing and intramolecular recombination. TherecE6 allele was then introduced into a set of DNA repair-deficient strains ofB. subtilis. The removal of DNA damage by therecF,addAaddB,recH,recL andrecP gene products is strictly dependent on an activerecE gene product (recE-dependent pathway). On the other hand, the increased sensitization to purine adducts in theuvrA42recE6 andpolA5recE6 strains suggests that such lethal lesions may be removed either by therecE-dependent or by therecE-independent pathway.     

Cloning of the polyketide synthase gene atX from Aspergillus terreus and its identification as the 6-Methylsalicylic acid synthase gene by heterologous expression

The geneCAL1 (also known asCDC43) ofSaccharomyces cerevisiae encodes the subunit of geranylgeranyl transferase I (GGTase I), which modifies several small GTPases. Biochemical analyses of the mutant enzymes encoded bycall-1, andcdc43-2 tocdc43-7, expressed in bacteria, have shown that all of the mutant enzymes possess reduced activity, and that none shows temperature-sensitive enzymatic activities. Nonetheless, all of thecall/cdc43 mutants show temperature-sensitive growth phenotypes. Increase in soluble pools of the small GTPases was observed in the yeast mutant cells at the restrictive temperature in vivo, suggesting that the yeast prenylation pathway itself is temperature sensitive. Thecall-1 mutation, located most proximal to the C-terminus of the protein, differs from the othercdc43 mutations in several respects. An increase in soluble Rholp was observed in thecall-1 strain grown at the restrictive temperature. The temperature-sensitive phenotype ofcall-1 is most efficiently suppressed by overproduction of Rholp. Overproduction of the other essential target, Cdc42p, in contrast, is deleterious incall-1 cells, but not in othercdc43 mutants or the wild-type strains. Thecdc43-5 mutant cells accumulate Cdc42p in soluble pools andcdc43-5 is suppressed by overproduction of Cdc42p. Thus, several phenotypic differences are observed among thecall/cdc43 mutations, possibly due to alterations in substrate specificity caused by the mutations.     

Restoration of respiratory competence in the yeastCandida utilis after somatic fusion withSaccharomyces cerevisiae

After somatic fusion between a mitochondrial mutant ofCandida utilis andSaccharomyces cerevisiae respiratory competent strain, the complex III of the respiratory chain seems to be restored. Fusion products, FP, synthesizing normal apocytochromeb were recovered and showed normal-shaped mitochondria along the cytoplasm as in theCandida utilis original, respiratory-competent strain.     

The effect of spo0 mutations on the expression of spo0A- and spo0F-lacZ fusions

Summary We have constructedspo0A-lacZ andspo0F-lacZ fusions with a temperate phage vector and have investigated howspo0 gene products are involved in the expression of each of these genes. The expression ofspo0A-lacZ andspo0F-lacZ was stimulated at about the time of cessation of vegetative growth in Spo⁺ cells. This stimulation ofspo0A-lacZ was impaired by mutations in thespo0B, D, E, F orH genes but was not affected by mutations in thespo0J orK genes. Similar results were obtained with thespo0F-lacZ fusion. The effect of thespo0A mutation onspo0A-lacZ expression was characteristic: thespo0A-directed β-galactosidase activity found during vegetative growth was significantly enhanced in thespo0A mutant. This result suggests thatspo0A gene expression is autoregulated being repressed by its own gene product. Another remarkable observation was the effect of thesof-1 mutation, which is known to be aspo0A allele; it suppressed the sporulation deficiency ofspo0B, spo0D andspo0F mutants. Thespo0A-lacZ stimulation, which is impaired by any one of thesespo0 mutations, was restored by the additionalsof-1 mutation.     

A numerical experiment on the determination of kinetic rate constants in the presence of diffusion

Two chemicals,A andB, are allowed to diffuse together and a reaction described by $$A + B\mathop \rightleftharpoons \limits_{K_{ - 1} }^{K_1 } C$$ is allowed to proceed. This system is described mathematically by a system of partial differential equations. A numerical procedure is presented to find the rate constants ofK ₁ andK _?1. A systematic analysis of the effects of errors is also presented.     

Electron donors and the quinone involved in dimethyl sulfoxide reduction inEscherichia coli

Escherichia coli can use dimethyl sulfoxide (DMSO) as an electron acceptor during anaerobic growth on the oxidizable substrate, glycerol. During growth, the DMSO is reduced to dimethyl sulfide (DMS). For the reduction of DMSO, NADH, formate, lactate, reduced benzyl viologen, reduced methyl viologen, and dithionite can serve as electron donors. The terminal reductase and the dehydrogenases linking the various electron donors to the electron transport chain were found to be membrane bound. Chlorate-resistant mutants (chl) were unable to grow and reduce DMSO. However, in the case of thechlD mutant, growth and DMSO reduction can be restored by growth in the presence of high concentrations of molybdate. Mutants ofE. coli blocked in menaquinone (vitamin K₂) biosynthesis—menB, menC, andmenD—were unable to grow with DMSO as an electron acceptor, even though the terminal reductase is present in these mutants. Both growth and DMSO reduction could be restored in these mutants by growth in the presence of the menaquinone intermediates,o-succinylbenzoate and 1,4-dihydroxy-2-naphthoate, depending on the metabolic block of the mutant. Thus menaquinone is involved in electron transport during DMSO reduction.     

Chromosome numbers and rust susceptibility as taxonomic criteria inRosaceae

BothNeuradoideae andChrysobalanoideae seem rust-free. TheSpiraeoideae andPomoideae are heavily susceptible toGymnosporangium rusts. While thePrunoideae resemble theRosoideae in being vulnerable to attacks ofPuccinia species, they are additionally susceptible toTranzschelia andThekopsora, whereas theRosoideae are characteristically afflicted byPhragmidium.—It is suggested (a) to treat theChrysobalanoideae as a separate family (Chrysobalanaceae), (b) to transferDryas from theRosoideae-Potentilleae-Dryadinae to theRosoideae-Cercocarpeae, and (c) to divide theRosoideae into two main groups of tribes: (i) the rust-freeKerrieae andCercocarpeae with x = 9, and (ii) the rust-susceptiblePotentilleae, Ulmarieae, Roseae andSanguisorbeae with x = 7.