The influence of anther cold pretreatments and donor plant genotypes on in vitro androgenesis in wheat (Triticum aestivum L.)

Cold pretreatments applied to excised anthers in liquid potato 2 medium proved to be unnecessary. Generally, cold pretreatments inhibited anther response and productivity as the duration was lengthened or as the pretreatment temperature was lowered. There were significant differences in response attributed to the anther donor genotype. Green and albino plants as well as roots only have been regenerated from the spring wheat cultivars Sinton, Neepawa, Pitic 62 and DW 50. Most plants were haploid.     

The role of balancing selection and overdominance in maintaining allozyme polymorphism

Three approaches to the estimation of the role of balancing selection in maintaining allozyme polymorphism are considered: 1) Analysis of the stationary distributions of allelic frequencies in a native subdivided population; 2) Comparison of the genotypic distributions at the early and late developmental stages in successive generations of the same population; 3) Analysis of the joint variability of monogenic and polygenic traits.The conclusion is drawn that allozyme polymorphism must not be regarded as a transient phase of molecular evolution but as its stationary phase. The mechanisms responsible for supporting such stability are discussed.     

Deterministic single-locus density-dependent selection

Density-regulated selection is considered for a single, multiallelic gene locus and separated generations. Characteristics resulting from the basic assumption that the average population fitness decreases with increasing density are derived. Under this assumption, it proves to be necessary to distinguish between regions of allelic frequencies which imply limited population growth, unlimited growth, or ultimate extinction when the population stays in the respective region. Particular attention is given to the investigation of the region of limited growth and the carrying capacity function defined on it. Relationships between and the average fitness (adaptive surface) in the non-density dependent model are explained.Besides stability properties of equilibrium points, more general characteristics concerning the asymptotic behavior of population trajectories are treated. In this context, the problems of sudden loss of alleles and of population extinction as a result of large fluctuations in density are discussed.     

The identification of a DNA polymorphism of the α fibrinogen gene,and the regional assignment of the human fibrinogen genes to 4q26-qter

Summary We have identified a common restriction fragment length polymorphism of the fibrinogen gene with the enzyme TaqI. This polymorphism is probably due to a single base change that creates or destroys a TaqI recognition site about 1000 basepairs from the 3 end of the fibrinogen géne. The frequency of the rare allele in 83 unrelated healthy individuals is 0.33. We have used in situ hybridisation of the fibrinogen cDNA to localise the gene on chromosome 4q29–31. We have confirmed this regional localisation by restriction fragment detection in a human x Chinese hamster somatic cell hybrid which contains a translocated human chromosome 4 with a breakpoint at 4q26. The , , and fibrinogen genes are all present on human chromosome 4q26-qter.     

Chemoenzymatic galactosialylation with integrated cofactor regeneration

As a precursor for the chemical synthesis of sialylated oligosaccharides, the trisaccharide glycoside Neu5Ac (2-8)Gal(1-4)GlcNAc(1-O)-pent-4-ene was synthesized starting from GlcNAc(1-O)-pent-4-ene, UDP-glucose andN-acetylneuraminic acid in a one pot reaction employing galactosyltransferase and (2-6)sialyl-transferase in a complete cofactor regeneration system.Abbreviations Neu5Ac N-acetylneuraminic acid - CMP-Neu5Ac cytidine 5-monophosphosialate - CMP cytidine 5-monophosphate - CDP cytidine 5-diphosphate - CTP cytidine 5-triphosphate - Gal galactose - GlcNAc N-acetylglucosamine - UDP uridine 5-diphosphate - UDP-Glc uridine-5-diphosphoglucose - UDP-Gal uridine-5-diphosphogalactose - PEP phosphoenolpyruvate     

Potential bias in inbreeding depression estimates when using pedigree relationships to assess the degree of homozygosity for loci under selection

A potential bias in estimation of inbreeding depression when using pedigree relationships to assess the degree of homozygosity for loci under selection is indicated. A comparison of inbreeding coefficients based on either pedigree or genotypic frequencies indicated that, as a result of selection, the inbreeding coefficient based on pedigree might not correspond with the random drift of allelic frequencies. Apparent differences in average levels of both inbreeding coefficients were obtained depending on the genetic model (additive versus dominance, initial allelic frequencies, heritability) and the selection system assumed (no versus mass selection). In the absence of selection, allelic frequencies within a small population change over generations due to random drift, and the pedigree-based inbreeding coefficient gives a proper assessment of the accompanying probability of increased homozygosity within a replicate by indicating the variance of allelic frequencies over replicates. With selection, in addition to random drift, directional change in allelic frequencies is not accounted for by the pedigree-based inbreeding coefficient. This result implies that estimation of inbreeding depression for traits under either direct or indirect selection, estimated by a regression of performance on pedigree-based coefficients, should be carefully interpreted.Deceased     

1∶1 and 2∶1 phase entrainment in a system of two coupled limit cycle oscillators

A model of a pair of coupled limit cycle oscillators is presented in order to investigate the extent of, and the transition between, 11 and 21 phase entrainment, a phenomenon exhibited by numerous diverse biological systems. The mathematical form of the model involves a flow on a phase torus given by two coupled first order nonlinear ordinary differential equations which govern the oscillators' phase angles (i.e. their respective positions around their limit cycles). The regions corresponding to 11 and 21 phase entrainment in an appropriate parameter space are determined analytically and numerically. The bifurcations occurring during the transition from 11 to 21 phase entrainment are discussed.     

Control of lectins in Triticum aestivum and Aegilops umbellulata by homoeologous group 1 chromosomes

Summary Each of the three genomes in hexaploid wheat controls the expression of a specific lectin in the embryo. The chromosomes which control their synthesis were determined using nullisomic-tetrasomic and inter-varietal chromosome substitution lines of Chinese Spring. All three wheat lectins were shown to be controlled by the homoeologous group 1 chromosomes. Using ditelosomic lines of Chinese Spring the lectin genes could be localized on the long arms of chromosomes 1A and 1D. Inter-specific addition and substitution lines of Aegilops umbellulata chromosomes to Chinese Spring indicated that chromosome 1U, which is homoeologous to the group 1 chromosomes of wheat, controls lectin synthesis.     

Actin filaments and microtubules in the preprophase band and phragmoplast of tobacco cells

Summary Treatment with lysine prior to fixation of tobacco BY-2 cells with formaldehyde improved the preservation of actin filaments in the cells and enabled us to observe both networks of actin filaments and microtubules in the same cells. By using this method, we observed that (1) actin filaments were present in the preprophase band; (2) the actin filaments in the preprophase band and phragmoplast were runnig in the same direction as the microtubules in their respective structures; (3) a cortical network of actin filaments was present throughout all stages of cell cycle.The present method did not preserve the cortical actin filaments in interphase cells. The procedure for staining microtubules destroyed them.Abbreviations EGTA Ethyleneglycol-bis(-aminoethyl ether)N,N,N,N-tetraacetic acid - PIPES Piperazine-N,N-bis(2-ethanesulfonic acid) - PMSF Phenylmethylsulfonyl fluoride - TLCK Na-p-tosyl-L-lysine chloromethyl ketone     

Expression of the β-subunit of β-conglycinin in seeds of transgenic plants

Soybean (Glycine max (L.) Merr.) seeds contain the storage protein -conglycinin, encoded by a multigene family. -Conglycinin consists of three subunits; , , and . A genomic clone for a -subunit of -conglycinin has been characterized by restriction-enzyme mapping and hybrid selected in-vitro translation followed by immunoprecipitation. In order to determine the developmental regulation of this -subunit gene, its expression was studied in seeds of transgenic petunia (Petunia hybrida) and tobacco (Nicotiana tabacum L.) plants. The -subunit expressed in seeds of petunia and tobacco was recognized by anti--conglycinin serum at a relative molecular mass of 53 000, equivalent to that of the native protein. Separation of the petunia-seed proteins by isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis showed that multiple isoelectric forms of the -subunit were produced. There was approximately a twofold variation in the accumulation of the -subunit protein in the mature seeds of transgenic petunia plants, each containing a single -subunit gene. However, the level of protein accumulation in mature seeds and the amount of -subunit mRNA in developing seeds was not correlated. Accumulation of the -subunit protein in transgenic seeds was less than the -subunit protein that accumulated in transgenic petunia seeds containing a single -subunit gene and less than the amount of the -subunit in mature soybean seeds which contain 8–13 -subunit genes. In transgenic tobacco plants, the accumulation of the -subunit protein in seeds was generally well correlated with the number of genes that were incorporated in the different transformants.Abbreviations kb kilobase - kDa kilodalton - M_r relative molecular mass - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis     

A model for population regulation with density- and frequency-dependent selection

The life-cycle of a species with separate generations is divided into a reproduction phase and a growing-up phase. In the reproduction phase we assume random mating and selection due to genotype differences in fecundity of the parents and viability of the offspring. During the growing-up phase we assume a (deterministic) death process in continuous time with death rates for the genotypes which increase linearly with the genotype population sizes.In the absence of genotype differences the model gives logistic population regulation. With genotype differences the model generalizes the usual separate generations selection patterns. In addition to these we exhibit cases with three polymorphic equilibria or with a stable cycle.     

Genetically determined differences in the response of alpha1-antitrypsin levels in human serum to typhoid vaccine

Summary Normal individuals and those homozygous and heterozygous for the gene which determines deficiency in serum ₁-antitrypsin (₁-at) were given an intravenous injection of typhoid vaccine. Following this injection the trypsin-inhibiting capacity of serum and the concentration of ₁-at increases markedly in normal individuals. Heterozygotes for the deficiency gene have an elevation of ₁-at and trypsin-inhibiting capacity of their serum but reach only 50 percent of the levels reached by homozygotes for the common gene (normal individuals). Their values are however temporarily in the range of normal men not given typhoid vaccine. Homozygotes for the deficiency gene show only a small elevation of ₁-at under these conditions. These findings demonstrate that (1) the increase in trypsin-inhibiting capacity of serum after an injection of typhoid vaccine is largely due to the elevation of the ₁-at concentration, and (2) the ₁-at deficiency gene inhibits the quantitative response of the ₁-at to such a stimulus.Supported in part by USPHS grant HE-06285 from the National Heart Institute.Some of these studies were carried out in the General Clinical Research Center Ward, U. C., FR-79, supported by the Division of Research Facilities and Resources, U.S. Public Health Service.     

Genetic differentiation amongst populations of the coral Pocillopora damicornis off southwestern Australia

An electrophoretic study of genetic differentiation amongst local populations of the reef-coral Pocillopora damicornis was used to group coral heads into units defined as the area of effective dispersal of a clone, and termed colonies. For reefs off southwestern Australia, colonies were usually under a few hundred metres in extent. Although most new recruits within a colony were derived asexually, sexually produced propagules acted to connect populations outside the boundaries of a colony. Such connections were weak, and allelic frequencies varied considerably over a few kilometres. The primary agent of genetic differentiation was suggested to be the small effective population size resulting from the asexual proliferation of a few genotypes at any site. The effective number of genotypes per colony was approximately six. Asexual reproduction appears also to limit gene flow and accentuate selection in this species.     

The origin and evolution of sexual reproduction up to the evolution of the male-female phenomenon

Summary Sexual reproduction is a composite, not a singular, phenomenon and as such can be subdivided into a number of componentsi.e. fusion, recombination, fission, and the male-female phenomenon. These components can evolve independently, though any evolutionary change in one component is likely to influence the future evolution of the other components. The ambiguity that surrounds the term sex due to a failure to recognise the composite nature of sexual reproduction has led to considerable confusion in past discussions of the evolution of the phenomenon. This paper considers the possible chronological interaction of the components of sexual reproduction both with each other and with the sequence of selective pressures that seem likely to have acted. This chronological approach is used to consider: the origin of sexual reproduction; the evolution of sexual reproduction in the common ancestor of the procaryotes and eucaryotes; the modification of the ancestral system in the procaryote line following the procaryote-eucaryote dichotomy; and the modification of the ancestral system in the eucaryote line up to the origin of the male-female phenomenon.It is suggested that the fusion and recombination of the first living organisms were chronological continuations of the fusion and recombination of complex organic molecules that led up to the origin of life. The evolution of the third major component of sexual reproductioni.e. fission (replication), by definition coincided with the origin of life. Initial selection on the components of sexual reproduction are likely to have been related to the optimum manifestations of size, complexity, diversity, multiplication, and distribution. Resultant early evolutionary trends are likely to have been: selective fusion between more-similar organisms; increase in number of fissions per fusion; and less recombination.The procaryote-eucaryote dichotomy is argued to have evolved in response to the increasing cellular problems of packing and replicating an increasing amount of hereditary material. The evolution of a single circular hereditary organelle in the procaryote line is argued to have led to the loss of total fusion and the specialisation of individuals into either donors or recipients. The donor-recipient phenomenon of procaryotes is directly analogous to the male-female phenomenon of eucaryotes and leads to parallel evolution due to sexual selection in both groups. In the eucaryote line the ancestral mechanism of sexual reproduction is argued to have persisted through, but to have been greatly modified by, the evolution of complex machinery (mitotic/meiotic) for the handling of multiple hereditary organelles at cell division and reduction division. The evolutionary modification of the ancestral system of sexual reproduction is suggested to have led in eucaryotes to the evolution of: the species phenomenon; allelic recombination; and the male-female phenomenon.     

Induction of Reactive Oxygen Species and Phytoalexins in Onion (Allium cepa) Cell Culture by Biotic Elicitors Derived from the Fungus Botrytis cinerea

Metabolites of a phytopathogenic fungus Botrytis cinerea Pers. were analyzed for the presence of biotic elicitors. Three groups of elicitors competent in inducing defense responses inAllium cepa cells were identified and partly purified. The recognition of the elicitor signal in onion cells was shown to elevate the concentration of reactive oxygen species (ROS), namely, superoxide anion-radical (O₂^{\overset{-}.}) and hydrogen peroxide (₂₂). The intensity of ROS release depended on chemical identity of elicitor and its concentration. The most active ROS production in onion cells was induced by a protein fraction isolated from the medium for fungus culturing. The carbohydrate elicitors extracted from the fungus cytoplasm and cell walls of mycelia were much less effective. The dynamics of ROS generation comprised two stages. The first stage represented fast and low-amplitude changes that peaked in 15 min after the elicitor treatment. The second stage was more durable and extensive; it occurred in 1.5–6 h after the treatment.     

Asexual reproduction and genetic determination of growth form in the coral Pavona cactus: biochemical genetic and immunogenic evidence

Summary Tissue grafting and electrophoresis were used to study the genotypic structure of a population of the scleractinian coral, Pavona cactus. Three growth forms were distinguished within one continuous population of this morphologically variable species. Both techniques provided evidence of localized asexual reproduction within each growth form, a result consistent with numerous field observations of naturally occurring fragments. A perfect association between clonal genotype and growth form was found in an electrophoretic survey of 80 colonies. 23 multi-locus genotypes were detected in the 80 colonies tested. All genotypically similar colonies had the same growth form, even where colonies were separated by 50 m. Although environmental gradients undoubtedly modify colony morphology, the high correlation between genotype and growth form suggests that major differences in colony morphology are genetically determined.Tissue grafting tests did not reliably identify all clones. Fusions developed between all electrophoretically indistinguishable colonies, consistent with the initial assumption that fusion between paired colonies would indicate selfrecognition. However, there was also one fusion in 20 pairings of electrophoretically different colonies. Although there was general agreement between the two techniques, the one inconsistent fusion suggests that caution should be exercised in the application of histocompatibility tests as bioassays for clonal population structure, and that electrophoresis is the more appropriate technique for this species.The ability of genotypes to dominate in intraspecific competitive interactions and to survive fragmentation was assessed. An intraspecific dominance hierarchy was identified among the 6 clones tested. Competition was highly asymmetrical between dominant and subordinate-ranking clones. Genotypes that were most successful in producing widespread clones were found to dominate intraspecific competitive interactions and had high rates of fragment survival. Offprint requests to: D.J. Ayre Contribution No 252 from the Australian Institute of Marine Science     

Evolutionary origin of expandable G-C-rich triplet repeat DNA sequences

A model explaining properties exhibited by fragile-X DNA systems arises from observations that time-dependent base substitutions are expressed at G-C sites but not at A–T sites (Biochem. Genet.32:383, 1994). [CGG]_n sequences are classified as most sensitive to evolutionary base substitution processes involving time-dependent populating of G-C sites with enol-imine states having enhanced stability. Increased density of these states in oocyte DNA would introduce a ground-state collapse double-helix of reduced energy that would inhibit strand separation by the replicase. Evolutionarily altered G in CGG triplets allows CGG to be transcribed as CTG, an initiation codon. And this will cause reinitiation of DNA synthesis, thereby adding additional CGG units to the collapsed double helix. This situation would not occur in slower-evolving male haploid DNA that replicates frequently.     

Nucleotide sequence of cytoplasmic 5S ribosomal RNA fromEuglena gracilis

Summary The nucleotide sequence of cytoplasmic 5S ribosomal RNA fromEuglena gracilis has been determined to be: G- _A ^C -G-U-A-C-G-G-C-C-A-U-A-C-U-A-C-C-G-G-G-A-A-U-A-C-A-C-C-U-G-A-A-C-C-C-G--U-C-G-A-U-U-U-C-A-G-A-A-G-U-U-A-A-G-C-C-G-G-G-U-U-A-G-G-C-C-C-A-G-U-U-A-G-U-A-C-U-G-A-G-U-G-G-G-C-G-A-C-C-A-C-U-U-G-G-G-A-A-C-A-C-U-G-G-G-U-G-C-U-G-U-A-C-G-C-U-Up. This RNA is 119 nucleotides long and the sequence of a probable tRNA-binding site is GAUU (position 41–44 from the 5-terminus), which is the same as that of a trypanosoma species,Crithidia fasci-culata. TheEuglena 5S rRNA has a pseudouridine residue at position 38 and 3-terminus is phosphorylated. The 5S rRNA sequence ofEuglena resembles those of several other protozoa and higher animals rather than plants.On leave from Department of Zoology, Hiroshima University, Hiroshima, Japan     

The E. coli cell surface: on the genetic organization of the tolPAB cluster

Summary Strongly polar mutations in the tolP AB cluster have been isolated using bacteriophage Mu. Complementation tests with these Mu-induced mutants indicated that tolP and tolA are co-transcribed in a clockwise direction, and that tolB is independently transcribed.In addition, the transduction behaviour of bacteriophage dtol-29 is described. This transducing phage appears to carry all of tolB and tolA, and terminate in tolP. With tolB and tolP recipients, dtol-29 transduced as expected (by complementation and recombination, respectively). With a tolA recipient, dtol-29 behaved anomalously. Transduction frequencies were reduced 20-fold (compared to a tolB recipient), and two distinct sizes of transduction colonies arose on the deoxycholate selection plates. Analysis of these colonies showed that all the large colonies arose by recombination-transduction, while all the small colonies arose by complementation-transduction. These observations are in agreement with the results obtained with the Mu-induced mutants; furthermore, they suggest that either a weak internal promoter exists between tolP and tolA, or that the tolA cistron on dtol-29 is under the control of gene Q.Research Fellow of the National Cancer Institute of Canada.     

An experimental test of the efficiency of family selection in chickens

Summary Responses to single trait selection on individual phenotype and sire-family mean phenotype for survivor's egg weight and rate of lay were measured for a single generation in 13 replicates. Each replicate-selection criterion-trait subclass consisted of eight sire families or 72 females measured and was reproduced from the best 25% of the families or individuals. The realized heritability of egg weight was 0.39 and that of rate of lay was 0.31, both of which were significantly greater than zero but not significantly different from the predicted values based on halfsib correlations in the base population.The standardized response to sire-family selection was less than the response to individual selection for both traits and the difference was significant for rate of lay (0.10; 0.31) but not for egg weight (0.22; 0.39). The predicted responses to sire-family selection were less than those for individual selection for both traits, and the observed responses to sire-family selection were not significantly different from the predicted values for either trait.These experimental results do not disagree with the theoretical expectations of the relative efficiencies of individual and sire-family selection.Journal paper no. 7479, Purdue University, Agricultural Experiment Station. This investigation was conducted as a part of the cooperative research of the NC-89 Regional Poultry Breeding Project entitled Nature and Utilization of Genetic Variation in Poultry Improvement