Phenotypic Variations in the Life History of Two Clones of Macrobiotus richtersi (Eutardigrada, Macrobiotidae)

A comparative study of life history traits of two clones (CDMr01 and CDMr02) of a triploid thelytokous apomictic population of the eutardigrade Macrobiotus richtersi has been carried out. Both clones were reared under the same lab conditions: temperature of 14 °C, photoperiod of 12 h/12 h (L/D), and nematodes ad libitum as food. Statistical analysis of the life history traits considered has indicated interclonal variability. The two clones were significantly different in the number of eggs per clutch (fertility), number of eggs laid per female per life span (fecundity), hatching percentage of eggs and hatching time. Similarities between clones have been observed with regard to life span, total number of ovipositions per life span, and age at first oviposition. In addition, significant differences in hatching time, with eggs hatched over a long period, were found within each clone. Interclonal variability in life history traits indicated the effects of genetic factors, whereas intraclonal variability reflected the effects of environmental factors. The evolutionary and adaptive significance of the life history phenotypic variations is discussed.     

Allozymes as aids for identification and differentiation of some Populus maximowiczii Henry Clonal varieties

Allozyme genotypes of nine Populus maximowiczii clonal varieties originating in China and Japan were determined for 35 loci coding for 12 enzyme systems in root tips. The interclonal variability was controlled by 13 polymorphic loci. Seven unique multilocus genotypes were identified. The unique genotypes differed from each other on an average of 6.73 loci. Principal Component (PC) 1 from the Principal Component Analysis of the clonal allozyme genotype data separated the clones into two distinct groups; one consisting of clones of Chinese and another clones of Japanese origins. Japanese clones of different sources were further differentiated by the PC 2. Allozyme genotypes were found to be useful for identification of clones and differentiation of their geographic origins.     

INTRA- AND INTERCLONAL COMPETITION IN THE CLEISTOGAMOUS GRASS AMPHIBROMUS SCABRIVALVIS

The relationship of differences in life history traits among genotypes to competitive ability is not well known for most clonal plants. It has been hypothesized that genetically identical clones will compete more intensively than genetically distinct clones. The perennial grass Amphibromus scabrivalvis, which produces basal corms and cleistogamous seeds enclosed by leaf sheaths, exhibits pronounced clonal growth via rhizome and ramet production. In a controlled greenhouse experiment, clones of four genotypes of this species were grown under three regimes: alone in the absence of competition, paired with a clone of the same genotype (intraclonal competition), and paired with a clone of a different genotype (interclonal competition). There were differences in some biomass measures and in ramet and corm production among the four genotypes grown in the absence of competition. All genotypes showed a significant reduction in total biomass under both intra- and interclonal conditions, indicating that competition had occurred. For three of four genotypes, biomass allocation to corm increased under competition, while allocation to cleistogamous seeds was constant or increased slightly. Although some genotypes in specific interclonal combinations were less affected by competition than in intraclonal combinations, there was no support for the contention that the effects of competition were more intense for genetically identical clones.     

Clone-specific immune reactions in a trematode-crustacean system

Variability of immune responses is an essential aspect of ecological immunology, yet how much of this variability is due to differences among parasite genotypes remains unknown. Here, variation in immune response of the crab, Macrophthalmus hirtipes, is examined as a function of experimental exposure to 10 clonal cercarial lineages of the trematode Maritrema novaezealandensis. Our goals were (1) to assess the variability of the host immune reaction elicited by 10 parasite clones, (2) to test if the heterozygosity-fitness correlation, whereby organisms with higher heterozygosities achieve a higher fitness than those with lower heterozygosities, applies to heterozygous parasites eliciting weak immune responses, and (3) to see how concomitant infections by other macroparasites influence the crab's immune response to cercariae. Parasite clones were distinguished and heterozygosities calculated using 20 microsatellite markers. We found that exposure to cercariae resulted in increased haemocyte counts, and that although interclonal differences in immune response elicited were detected, parasite heterozygosity did not correlate with host immune response. Additionally, the presence of other pre-existing parasites in hosts did not influence their immune response following experimental exposure to cercariae. Overall, the existence of variability in immune response elicited by different parasite clones is promising for future ecological immunology studies using this system.     

Methylation patterns in the gene for the alpha subunit of chorionic gonadotropin are inherited with variable fidelity in clonal lineages of human fibroblasts.

Cytosine methylation in DNA of an endogenous single-copy gene locus, the alpha-subunit of human chorionic gonadotropin (alpha-hCG), was assessed in a mass culture and individual clonal lineages of human diploid fibroblasts. Progressive hypomethylation at -CCGG- sites in this gene occurred during the replicative lifespan of the mass culture and was shown to arise randomly during clonal expansion. Thus, some clones and subclones lost -CCGG- methylation in the alpha-hCG gene region while others maintained essentially complete methylation. These data indicate significant interclonal variability in the fidelity with which DNA methylation is transmitted in an endogenous gene.     

Visible and "cryptic" segregation of parental chromosomes in embryonic stem hybrid cells

Chromosome segregation of the parental chromosomes was studied in 20 interspecific hybrid clones obtained by fusion of Mus musculus embryonic stem cells with Mus caroli splenocytes. FISH analysis with labeled species specific probes and microsatellite markers was used for identification of the parental chromosomes. Cytogenetic analysis has shown significant intra- and interclonal variability in chromosome numbers and ratios of the parental chromosomes in the hybrid cells: six clones contained all M. caroli chromosomes, nine clones showed moderate segregation of M. caroli chromosomes (from 1 to 7), and five clones showed extensive loss of M. caroli chromosomes (from 12 to complete loss of all M. caroli autosomes). Both methods demonstrated "cryptic" segregation of the somatic partner chromosomes. For instance, five clones with near-tetraploid chromosome sets contained only few M. caroli chromosomes (from 1 to 8). The data obtained suggest that the tetraploid chromosome set per se is not a sufficient criterion for conclusion on the absence of chromosome loss in the hybrid cells. Note that "cryptic" chromosome segregation occurred at a high frequency in the examined hybrid clones. Thus, "cryptic" segregation should be borne in mind for assessing pluripotency and genome reprogramming of embryonic stem hybrid cells.     

Comparison of Nicotiana tabacum and Nicotiana nesophila hybrids produced by ovule culture and protoplast fusion

Summary Somatic hybrids were produced between Nicotiana tabacum and N. nesophila, two species incapable of conventional sexual hybridization. Sexual hybrids, though, could be produced between these two species by using ovule culture only when N. nesophila was female. Clones of somatic hybrids were compared with sexual hybrids. Statistically significant variation was observed between clones, but not between sexual hybrids, for pollen viability, flower morphology, leaf morphology, and trichome density. As all clones of somatic hybrids have 96 chromosomes, the variability could not be explained by interclonal variation in chromosome number. Variation between somatic hybrids could be the result of cytoplasmic segregation or recombination, mitotic recombination or small chromosomal rearrangements prior to plant regeneration. Variation between clones could be exploited as these interspecies hybrids are now being used to incorporate disease resistance into cultivated tobacco.     

Morphological, developmental and electrophoretic variation within and between obligately apomictic Taraxacum species

Interclonal variation in six morphometric characters and in the onset and rate of capitulum production, together with inter-and intra-clonal variation in esterase zymograms was examined in each of the two obligately apomictic species, Taraxacum pseudohamatum (section Hamata ) and T. unguilobum (section Naevosa ). Interclonal and interspecific variation in esterase zymograms was also examined within and between seven additional obligately apomictic species within the section Hamata. Within T. pseudohamatum , interclonal variation in the morphometric and electrophoretic characters was extensive; no variation was recorded in the onset and rate of capitulum production. Within T. unguilobum , interclonal variation in the morphometric characters and in the onset and rate of capitulum production was recorded; five of the six clones electrophoretically assayed shared the same esterase zymogram but one of these clones contained an individual which exhibited an aberrant zymogram. Within the section Hamata , considerable variation in esterase zymograms was recorded; this variation was both inter-and intra-specific although one zymogram predominated, being represented in each species and in 65% of the clones.     

Intraclonal and interclonal differences of ploidy in transplantable rat rhabdomyosarcoma RA-2 detected using DNA flow cytometry

DNA contents were determined using flow cytometry in cells of 71 clones of the rat transplantable rhabdomyosarcoma passed through more than 70 cycles of selection for increased malignancy. 58 clones (82%) consisted of cells with diploid DNA content, whereas 13 clones (18%) displayed not only diploid, but also near tetraploid cell subpopulations, the latter covering from 20 to 60% of the total number of cells measured. The data obtained show the existence of intra- and interclonal karyotypic heterogeneity in populations of tumor clonogenic cells.     

Influence of temperature and cell size on the division rate and chemical content of the diatom Chaetoceros curvisetum Cleve

Cell size and temperature influenced the division rate and chemical content of the diatom Chaetoceros curvisetum Cleve when grown at 15, 20 and 25°C in nutrient replete media. Cell-size dependent trends of division rate in individual clones changed with temperature in a complex fashion. Considerable interclonal variability in division rate within a restricted range of cell sizes was also found. Cellular levels of carbon, nitrogen, protein, chlorophyll a, and silicon were linearly related to cell size. Cellular levels of carbon and chlorophyll per unit volume and silicon per unit surface area changed with temperature. No temperature effect on cellular levels of nitrogen and protein was found.     

Caractères taxinomiques et spécificité dans le genreAsplanchna (Rotifère)

A comparative analysis of eight characters in two clones ofA. brightwelli and a clone ofA. intermedia, fed withParamecium orBrachionus, is made to determine the intraclonal, interclonal and interspecific variations in this complex of species.The trophi morphology appears the most species specific criterium whereas the other features (size of males, diameter and ornamentation of resting eggs, nuclear number in syncytial glands) show variability, either genetic or related to environmental conditions. The variation observed in the ornamentation of resting eggs and in the nuclear number of vitellogen was distributed along a continuous gradient.

     

Interclonal differences, plasticity and trade-offs of life history traits of Cyperus esculentus in relation to water availability

Cyperus esculentus is an exotic clonal (or pseudoannual) weed in Japan, and its range is steadily increasing. To investigate its interclonal variation and phenotypic plasticity in response to water availability, five clones of C. esculentus , collected from different sites in Japan, were grown singly in pots placed outdoors under dry and wet conditions. All the traits examined showed considerable variation among the five clones. However, two clones from Tochigi were similar to each other; thus, they might have originated from the same founder population. The clone from Ibaraki was quite different from the others. Therefore, it is suggested that the Japanese populations of C. esculentus might have resulted from multiple introductions of genotypes from geographically separated and, hence, genetically differentiated, source populations. All the clones also showed considerable plasticity in response to water availability. Clones with a larger ramet number had a greater plasticity, whereas tuber size was invariant across water treatments. Highly plastic traits had generally low interclonal variation in plasticity. All the clones had high productivity and produced more ramets and tubers under wet conditions than under dry conditions. Moreover, water availability could partially regulate the mode of its reproduction; wet conditions favored tuber production (vegetative propagation) while dry conditions favored sexual reproduction. A number of trade-offs occurred between the traits of clonal growth, storage and sexual reproduction, indicating that allocation among the competing functions/organs is mutually exclusive in plants. The results obtained here suggest that C. esculentus is more likely to invade wet habitats than dry habitats.     

Molecular and dendrochronological analysis of natural root grafting in Populus tremuloides (Salicaceae)

Trembling aspen (Populus tremuloides) is a clonal tree species, which regenerates mostly through root suckering. In spite of vegetative propagation, aspen maintains high levels of clonal diversity. We hypothesized that the maintenance of clonal diversity in this species can be facilitated by integrating different clones through natural root grafts into aspen's communal root system. To verify this hypothesis, we analyzed root systems of three pure aspen stands where clones had been delineated with the help of molecular markers. Grafting between roots was frequent regardless of their genotypes. Root system excavations revealed that many roots were still living below trees that had been dead for several years. Some of these roots had no root connections other than grafts to living ramets of different clones. The uncovered root systems did not include any unique genotypes that would not occur among stems. Nevertheless, acquiring roots of dead trees helps to maintain extensive root systems, which increases the chances of clone survival. Substantial interconnectivity within clones as well as between clones via interclonal grafts results in formation of large genetically diverse physiological units. Such a clonal structure can significantly affect interpretations of diverse ecophysiological processes in forests of trembling aspen.     

Genetic Diversity among Different Clones of the Gynogenetic Silver Crucian Carp, Carassius auratus gibelio, Revealed by Transferrin and Isozyme Markers

Genetic diversity among four clones (A, D, E, F) of gynogenetic silver crucian carp was studied using transferrin and isozymes in the blood as markers. Of the five proteins investigated, three (transferrin, esterase and superoxide dismutase) indicated polymorphism and eight polymorphic loci were detected. These loci were probably encoded by codominant alleles and their inheritance patterns were analyzed. Intraclonal homogeneity and interclonal heterogeneity were observed in these clones, which allowed us to infer the clonal nature and evolutionary relationship between them. Clonal diversity in this population of silver crucian carp in China was also compared with data reported from gynogenetic crucian carp in Germany.     

Somatic embryogenesis and somaclonal variation in Norway spruce: morphogenetic, cytogenetic and molecular approaches

Four embryogenic clones of Norway spruce have been subcultivated and observed over several years to determine the evolution of production of mature embryos and to assess the quality of the embryos produced. A wide range of intraclonal quantitative and qualitative variability has been observed within this production. Certain morphologic deviations appeared at the immature stage and after maturation, such as immature embryos with a diffuse organization, complete or part albino mature embryos or acclimated somatic seedlings comparable to dwarf mutants. All of these phenotypic variations could be the result of a modification of the genome itself or of only the expression of the genome. Two approaches, chromosome counting and RAPD (random amplified polymorphic DNA), were chosen for their capacity to detect genotypic variations: respectively, genomic and chromosomic or genic mutations. The cytogenetic approach revealed, for the first time in this species, three cases of mutated acclimated somatic plants: one totally trisomic and two chimeras with trisomic buds and diploid roots. Other cases of 5-year-old trisomic, double trisomic, tetraploid or mixoploid embryogenic masses were also detected. The molecular approach (RAPD) revealed no somaclonal variation despite the large sample of DNA and primers used and the important interclonal variation observed. Received: 9 June 1996 / Accepted: 21 June 1996     

Fine-scale adaptation in a clonal sea anemone

Local adaptation in response to fine-scale spatial heterogeneity is well documented in terrestrial ecosystems. In contrast, in marine environments local adaptation has rarely been documented or rigorously explored. This may reflect real or anticipated effects of genetic homogenization, resulting from widespread dispersal in the sea. However, evolutionary theory predicts that for the many benthic species with complex life histories that include both sexual and asexual phases, each parental habitat patch should become dominated by the fittest and most competitive clones. In this study we used genotypic mapping to show that within headlands, clones of the sea anemone Actinia tenebrosa show restricted distributions to specific habitats despite the potential for more widespread dispersal. On these same shores we used reciprocal transplant experiments that revealed strikingly better performance of clones within their natal rather than foreign habitats as judged by survivorship, asexual fecundity, and growth. These findings highlight the importance of selection for fine-scale environmental adaptation in marine taxa and imply that the genotypic structure of populations reflects extensive periods of interclonal competition and site-specific selection.     

Triterpene methyl ethers in leaf waxes of Saccharum and related genera

The triterpene methyl ethers in the leaf waxes of over 80 clones of Saccharum officinarum, S. edule, S. robustum, S. spontaneum and a limited number of related species were compared as possible chemotaxonomic markers by GLC. The principal components were arundoin, crusgallin and cylindrin. The overall interspecific variation was small, but arundoin was particularly characteristic of S. officinarum. However, each species showed marked interclonal variation, which was related to chromosome numbers and geographical origin. Most S. spontaneum clones from India were atypical containing no triterpene methyl ethers.     

Predator-induced morphological defence in ciliates: interclonal variation for sensitivity to the inducing factors

This study brings the first evidence for the interclonal variation in the expression of predator-induced morphological defence in ciliates. The degree of response to the ciliate predator Stylonychia mytilus was compared in 18 clones of Euplotes octocarinatus . Although all the examined clones were inducible, they attained similar level of the response at a different "dose" of inducing factor. One of the clones maintained the "induced form" permanently, i.e. in the absence of any predator. Furthermore, comparing the effects of two different predators S. mytilus and a turbellarian Stenostomum sp. revealed differences among closely related clones in their relative responses to these predators. The degree of response in a given clone to one predator species did not preclude its response level to the other one suggesting that the expression of the defence to different predators might change independently. Our findings indicate that there is genetic variation for sensitivity to the inducing factors in Euplotes and hence the possibility for selection on this variation.     

Functional analysis of macrophage hybridomas. I. Production and initial characterization

A series of macrophage hybridomas were generated by fusion of splenic adherent cells with P388D1 tumor cells. Forty-two cell lines were established, and each was cloned by limiting dilution. Six clones that exemplified the spectrum of macrophage heterogeneity were selected for further analysis. Qualitative and quantitative differences in phenotype and functional activity were noted. Some clones constitutively expressed Ia antigens, whereas others only expressed detectable levels of Ia after lymphokine activation. The level of antigen-presenting activity generally correlated with the level of Ia expression. Furthermore, interclonal differences were noted in the levels of receptor-mediated phagocytosis and IL 1 secretion. Generally, the hybridoma clones maintained stable phenotypic and functional properties during approximately 1 yr of continuous in vitro culture. These cloned hybridoma cell lines represent a useful resource to analyze macrophage biology and to dissect structure and function relationships.     

Phenotypic heterogeneity in a panel of infectious molecular human immunodeficiency virus type 1 clones derived from a single individual.

Previously, we and others have demonstrated a relation between the clinical course of human immunodeficiency virus type 1 (HIV-1) infection and biological properties of HIV-1 variants such as replication rate, syncytium-inducing (SI) capacity, and cytotropism. For the molecular analysis of the biological variability in these properties, we generated a panel of phenotypically distinct yet genetically highly homologous infectious molecular clones. These clones were derived from HIV-1 isolates, mostly recovered by direct clonal isolation, from a single individual in whom a transition from non-SI to SI isolates had been identified over time. Of 17 molecular clones tested, 8 were infectious. The clones exhibited differences in SI capacity and T-cell line tropism. Their phenotypes corresponded to those of their parental isolates, formally demonstrating that biological variability of HIV-1 isolates can be attributed to single molecular clones. With these clones we demonstrated that SI capacity and tropism for the H9 T-cell line, almost invariably coupled in primary HIV-1 isolates, are discernible properties. Also different requirements appeared to exist for H9 and Sup T1 cell line tropism. We obtained evidence that T-cell line tropism is not caused by differences in level of HIV-1 expression but most probably is restricted at the level of virus entry. Restriction mapping of four clones with divergent phenotypes revealed a high degree of nucleotide sequence homology (over 96.3%), indicating the usefulness of these clones for the tracking of genetic variability critical for differences in biological phenotype.