A Study of Odd Mating-Type Determination Factor by Transfer of Macronuclear Karyoplasm in PARAMECIUM TETRAURELIA

The unique feature of the "B system" of mating-type determination found in Paramecium tetraurelia is the existence of a cytoplasmic difference between odd (O) and even (E) cells created and maintained by the action of their macronuclei. Thus far, the presence of a determining factor that controls the differentiation of the developing zygotic macronucleus for O mating type has not been verified. Results of crosses between cells of differing clonal age and complementary mating type suggest that, for one to two fissions after autogamy, O cells produce some factor that determines the gametic nucleus (micronucleus) as mating type O. Direct evidence for the production of O-determining factor by the young O macronucleus was obtained by transplanting young O macronuclear karyoplasm (a part of the macronucleus) into E cells: 32-35% of E exautogamous clones transformed to O; transformation of E exautogamous clones to O reached as high as 72% by transfer of young O macronuclear karyoplasm from a conjugant, 3-4 hr after mixing. This indicates that O determinants produced by the O macronucleus can also act during the sensitive period of development of the new macronucleus. These O-determining factors may be produced or activated at the sexual stage and then decrease in activity in subsequent fissions after new macronuclear reorganization.     

Mating-Type Inheritance and Maturity Times in Crosses between Subspecies of TETRAHYMENA PIGMENTOSA

Subspecies 6 and 8 of T. pigmentosa (formerly syngens 6 and 8 of T. pyriformis) share a mating-type system controlled by three alleles with "peck-order" dominance at a single locus. The system is apparently closed and limited to three mating types that are homologous, but not identical, in the subspecies. These relationships are reflected in new mating-type designations.—The viability in some intersyngenic crosses is excellent, and the inheritance of major mating types in first-generation hybrids and their progeny follows the pattern of subspecies 8.—The period of immaturity is shorter than that previously reported for subspecies 8, with 50% of the subclones maturing between 46 and 100 fissions after conjugation. Maturity curves are generally sigmoid, but some are apparently biphasic. The onset of maturity in triplicate sublines from the same synclone is usually highly correlated.     

Mutants of Sexual Maturity in PARAMECIUM CAUDATUM Selected by Erythromycin Resistance

Cells of Paramecium caudatum, syngen 3 usually become sexually mature about 50 fissions after conjugation. In order to study the genetic mechanisms that control fission-dependent expression of maturity, an attempt was made to obtain early mature mutants by treatment with N-methyl-N'-nitro-N-nitro-soguanidine. A new cytoplasmic marker, erythromycin resistance, was used to eliminate nonconjugant and macronuclear regeneration clones. Twenty early mature clones were obtained from five different mutagenized cultures. Three of them were genetically analyzed by crosses to wild-type stocks. The results show all three mutants to be controlled by incompletely dominant genes, i.e., the homozygotes became mature 20-25 fissions and the heterozygotes 15 fissions earlier than the wild-type clones. At least two different loci are suggested for the early maturity.     

Analysis of germinal aging in Paramecium caudatum by micronuclear transplantation

The role of the micronucleus in the age-dependent increase in mortality after conjugation in Paramecium has been investigated using micronuclear transplantation. The clone of Paramecium caudatum used for this study had a lifespan of about 750 fissions. In this clone, the fission rate began to decrease about 450 fissions after conjugation. Mortality after selfing conjugation also began to appear at about 450 fissions and gradually increased with clonal age. Cells at about 650 fissions showed 10–70% survival after selfing conjugation but when their micronuclei were transplanted into amicronucleate cells of about 450 fissions, the progeny survival increased to 70–90%. When micronuclei from cells 700–750 fissions old were transplanted into amicronucleate cells of 100–150 fissions, however, increase in progeny survival was very rare. The results indicate that micronuclei in cells up to the age of 650 fissions can function normally if the cytoplasmic environment is young.     

Novel features of computer-simulated clonal life of Paramecium caudatum

The clone of the ciliated protozoan Paramecium caudatum has the immaturity period of about 60 fissions and the lifespan of about 600 fissions. These life cycle figures have been depicted through laboratory experiments that allow continuous cell divisions for hundreds, which never occur in nature. We here constructed the nature-mimicking model culture that alternated the log- and stationary phases to allow conjugation, and computer-simulated the age structure modifying parameters such as cell distributions to start the culture, fission rates, death rates, immaturity periods, probabilities of conjugation, proportions of transplantation and so on. The average and maximum ages in the culture after thousands of alternations were converged to 43±2 and 140±5 fissions, respectively, when parameters for the immaturity period and the maximum clonal lifespan were set at 60 and 600 fissions. This result explains why cells collected in nature are usually young and vigorous. The average and maximum ages proportionally prolonged as the immaturity period was prolonged, as reported true for species of the ciliate. These results indicate the validity of our simulation. The average and maximum ages remained unchanged when the initial condition for starting the culture was changed from two complementary mating-type cells to a population with a quadratic-function distribution, and when the fission rate at the log-phase and the death rate at the stationary phase were modified for older ages. The average and maximum ages changed slightly when either the conjugation rate or the proportion of transplantation was somewhat lowered. Although they changed considerably when such parameters as the immaturity period, conjugation rate and death rate were extremely modified, no clones with the age over 230 fissions appeared in any simulations. These results indicate the robustness of the model, which provides us with fresh insight into the structural system of the clonal lifespan of P. caudatum in nature.     

New approaches to the problem of macronuclear assortment

Macronuclear assortment is a vegetative (somatic) process in the ciliate protozoan genus Tetrahymena which leads to the production of “homozygous” phenotypes from sexually-produced heterozygotes. The appearance of pure types follows a model of 45 randomly distributed and replicating subunits in the polyploid macronucleus. Models for the organization of macronuclear chromatin must explain the following: (1) haploid genomes are distributed at each fission; (2) not all loci assort; (3) different loci begin assortment at different times following conjugation. Therefore, either haploid subunits distributed at fission must frequently exchange parts (by somatic recombination, gene conversion, or progressive chromosome fragmentation), or they may actually disintegrate between fissions. Some of these possibilities can be distinguished by considering the kinetics of the appearance of pure types at two or more loci simultaneously. Conditions which give optimum discrimination between hypotheses of limited recombination and complete scrambling (by disintegration of haploid units between fissions) have been developed. A method for determining the number of assorting subunits in very young macronuclei is also given.     

The impact of group fissions on genetic structure in Native South America and implications for human evolution

In a series of publications beginning in the 1960s, Neel and colleagues suggested that genetically nonrandom, or "lineal", population fissions contributed to genetic structure in ancient human groups. The authors reached this conclusion by studying the genetic consequences of village fissions among the Yanomamo, a Native South American group thought to have been relatively unaffected by European contact and, therefore, representative of the human past. On the basis of ethnographic accounts and pedigree data, they further concluded that patrilineal relationships were particularly important in shaping the genetic structure of villages following fissions. This study reexamines the genetic consequences of village fissions using autosomal STRs, Y-chromosome STRs, and mitochondrial DNA sequences collected from large samples of individuals from multiple Yanomamo villages. Our analyses of the autosomal STRs replicate the previous finding that village fissions have produced substantial genetic structure among the Yanomamo. However, our analyses of Y-chromosome STRs and mtDNA d-loop polymorphisms suggest that other population processes, including village movements, inter-village migration, and polygynous marriage, affect genetic structure in ways not predicted by a simple model of patrilineal fissions. We discuss the broader implications of population fissions for human evolution and the suitability of using the Yanomamo as a model for the human past.     

Effects of fluctuating temperature and immersion on asexual reproduction in the intertidal sea anemone Hauplanella luciae (Verrill) in laboratory culture

Haliplanella luciae (Verrill) was maintained in laboratory cultures according to a 2 × 2 factorial design. The factors were 1) constant immersion or continuously alternating periods of 6 h immersion followed by 6 h emersion and 2) 18 °C constant temperature or 24 °C during one 6-h emersion period each day and 18 °C for the remaining 18 h. Significantly different numbers of fissions were observed between both the two immersion and two temperature conditions. A negative correlation was found between the number of fissions and the total dry weight of tissue per bowl. There was a different distribution of individual weights in the two immersion conditions. Some instances of pedal laceration were observed.     

Chromosomal Diversity and Karyotype Evolution in South American Macaws (Psittaciformes,Psittacidae)

Most species of macaws, which represent the largest species of Neotropical Psittacidae, characterized by their long tails and exuberant colours, are endangered, mainly because of hunting, illegal trade and habitat destruction. Long tailed species seem to represent a monophyletic group within Psittacidae, supported by cytogenetic data. Hence, these species show karyotypes with predominance of biarmed macrochromosomes, in contrast to short tailed species, with a predominance of acro/telocentric macrochromosomes. Because of their similar karyotypes, it has been proposed that inversions and translocations may be the main types of rearrangements occurring during the evolution of this group. However, only one species of macaw, Ara macao, that has had its genome sequenced was analyzed by means of molecular cytogenetics. Hence, in order to verify the rearrangements, we analyzed the karyotype of two species of macaws, Ara chloropterus and Anodorhynchus hyacinthinus, using cross-species chromosome painting with two different sets of probes from chicken and white hawk. Both intra- and interchromosomal rearrangements were observed. Chicken probes revealed the occurrence of fusions, fissions and inversions in both species, while the probes from white hawk determined the correct breakpoints or chromosome segments involved in the rearrangements. Some of these rearrangements were common for both species of macaws (fission of GGA1 and fusions of GGA1p/GGA4q, GGA6/GGA7 and GGA8/GGA9), while the fissions of GGA 2 and 4p were found only in A. chloropterus. These results confirm that despite apparent chromosomal similarity, macaws have very diverse karyotypes, which differ from each other not only by inversions and translocations as postulated before, but also by fissions and fusions.     

GENESIS: genome evolution scenarios

Summary: We implemented a software tool called GENESIS for threedifferent genome rearrangement problems: Sorting a unichromosomalgenome by weighted reversals and transpositions (SwRT), sortinga multichromosomal genome by reversals, translocations, fusionsand fissions (SRTl), and sorting a multichromosomal genome byweighted reversals, translocations, fusions, fissions and transpositions(SwRTTl). Availability: Source code can be obtained by the authors, oruse the web interface http://www.uni-ulm.de/in/theo/research/genesis.html Contact: simon.gog{at}uni-ulm.de Associate Editor: Chris Stoeckert     

Chromatin morphology and cytokinesis in pleurocapsalean cyanobacteria

Chromatin morphology and cytokinesis are described in the cyanobacterium Pleurocapsa sp. CALU 1126 capable of multiple fission (multiple reproduction of the mother cell, the macrocyte, with the formation of unique reproductive cells, the baeocytes by means of differential interference contrast (DIC) and fluorescence microscopy). Two kinds of chromatin behavior in the cell cycle are revealed, including (1) the formation of numerous chromatin areas prior to their compartmentalization by multiple fission and (2) chromatin condensation in the phase of binary fission and chromatin decondensation in growth period. The cytokinetic essence of multiple fission is shown to consist of successive binary fissions of the macrocyte, while in between, the mother cells (prebaeocytes) do not grow.     

A comparative banding analysis of chromosomes in three species of lemurs (Primates,Lemuridae)

Banded karyotypes were compared in 3 species of lemurs,Lemur catta (2n=56),L. f. fulvus (2n=60) andL. mongoz (2n=60). The karyotypes of the latter 2 species were indistinguishable from each other in both Q-band and G-band patterns, except thatL. mongoz had an unusually large Y chromosome. The karyotypic differences betweenL. catta and the other 2 species were mainly explained by centric fusions (or fissions) and pericentric inversions. Contrary to the general similarity in Q-band and G-band patterns, the C-band patterns were highly variable among the 3 species. All chromosomes ofL. fulvus had a distinctive C-band in the centromeric region, whileL. mongoz had only a few chromosomes with an apparent C-band.L. catta was remarkable by showing interstitial and terminal C-bands in some elements, in addition to the centromeric band which was observed in about 20 pairs.     

Recombination and Assortment in the Macronucleus of TETRAHYMENA THERMOPHILA: A Theoretical Study by Computer Simulation

The compound nature of the macronucleus of Tetrahymena thermophila presents multiple opportunities for recombination between genes on the same macronuclear chromosome. Such recombinants should be detectable through their assortment at subsequent amitotic macronuclear divisions. Thus, a macronucleus that is initially AB/ab should produce recombinant assortees of the genotypes Ab/aB. Computer simulation shows that, when the recombination frequency is two or fewer times per cell cycle, recombinant assortees are produced at experimentally measurable frequencies of less than 40%. At higher recombination frequencies, linked genes appear to assort independently. The simulations also show that recombination during macronuclear development can be distinguished from recombination in subsequent cell cycles only if the first appearance of recombinant assortees is 100 or more fissions after conjugation. The use of macronuclear recombination and assortment as a means of mapping macronuclear genes is severely constrained by the large variances in assortment outcomes; with experimentally small sample sizes, such mapping is impossible.     

New insights into sex chromosome evolution in anole lizards (Reptilia,Dactyloidae)

Anoles are a clade of iguanian lizards that underwent an extensive radiation between 125 and 65 million years ago. Their karyotypes show wide variation in diploid number spanning from 26 (Anolis evermanni) to 44 (A. insolitus). This chromosomal variation involves their sex chromosomes, ranging from simple systems (XX/XY), with heterochromosomes represented by either micro- or macrochromosomes, to multiple systems (X₁X₁X₂X₂/X₁X₂Y). Here, for the first time, the homology relationships of sex chromosomes have been investigated in nine anole lizards at the whole chromosome level. Cross-species chromosome painting using sex chromosome paints from A. carolinensis, Ctenonotus pogus and Norops sagrei and gene mapping of X-linked genes demonstrated that the anole ancestral sex chromosome system constituted by microchromosomes is retained in all the species with the ancestral karyotype (2n?=?36, 12 macro- and 24 microchromosomes). On the contrary, species with a derived karyotype, namely those belonging to genera Ctenonotus and Norops, show a series of rearrangements (fusions/fissions) involving autosomes/microchromosomes that led to the formation of their current sex chromosome systems. These results demonstrate that different autosomes were involved in translocations with sex chromosomes in closely related lineages of anole lizards and that several sequential microautosome/sex chromosome fusions lead to a remarkable increase in size of Norops sagrei sex chromosomes.     

Group Fission in Barbary Macaques (Macaca sylvanus) at Affenberg Salem

We analyzed eight group fissions occurring during a 20-year period in three groups of a free-ranging provisioned Barbary macaque population. The founder group fissioned four times within 3.5 years after transfer to the enclosure, indicating that external factors—new environment, more space, absence of other groups—facilitated group fissions. Two groups resulting from these fissions, split twice within 2.5 and 1 years, respectively, many years later. The process of fissioning lasted from a few months to almost 2 years. Fissions were preceded by peripheralization/subgrouping of mainly young adult males (8-10 years old), suggesting that male competition was the primary force for the fissions. The males were joined by middle- to low-ranking but not the lowest-ranking females. The resulting new groups were usually smaller than the groups in which the former -matriline—old groups—stayed, and they were also more variable in size and sex ratio, suggesting that variable numbers of surplus individuals were expelled during fission. Mean adult sex ratios were similar in both groups after fission, indicating that the competitively superior males in the old groups (groups + -matriline) could not increase their breeding opportunities. Female kin, even of large matrilinies, almost always stayed together during fission. Natal males strongly preferred to join the old groups, and this preference was most pronounced in juveniles and subadults. Hence, most natal males stayed with maternally related females, i.e., remained true natal males, if the females stayed in old groups. They were separated from female kin, i.e., became seminatal, if the females joined the new groups. These seminatal males did not differ from natal males with respect to matrilineal rank, but they had more female relatives, above all more close relatives (sisters), indicating that avoidance of mating with maternal kin was important for group choice. Despite joining the same group as female kin during fissioning, breeding opportunities of natal males (ratio of unrelated females/male) were not less than that of their seminatal peers, because natal males had fewer female relatives. Only a minority of both groups of males would have done better by joining the alternative group. Paternal relatives were distributed during fission by chance, and loss of patrilinies was therefore much less pronounced. We conclude that the rules governing social relationships among Barbary macaque males are less apt to cope with the high number of males resulting from provisioning, whereas the rules regulating social relationships of females living in a nepotistic, female-bonded society are very robust in this respect.     

Reversal and convergence in marsupial chromosome evolution

The karyotypes of marsupial species are characterized by their relatively low number of chromosomes, and their conservation. Most species have diploid numbers lying between the two modes, 2n = 14 and 2n = 22, but the karyotype of Aepyprymnus rufescens is exceptional in containing 2n = 32 chromosomes. Many differences in diploid number between marsupial species can be accounted for by particular fissions and fusions, which are easy to detect because of the low numbers of chromosomes in each karyotype. This should be a system in which it is possible to detect reversals and repeated chromosome rearrangements. We have used chromosome-specific paints derived from A. RUFESCENS to compare the karyotypes of eight marsupial species, representing closely and distantly related taxa, to trace chromosome change during evolution, and especially to detect reversals and convergence. From these and other painting comparisons, we conclude that there have been at least three reversals of fusions by fissions, and at least three fusions or fissions that have occurred independently in different lineages.     

Propagation of cortical differences in tetrahymena

Progeny clones were derived from crosses arranged so that the number of ciliary meridians (corticotype) was unusually high in one partner, and normal in the other. An analysis of the propagation of corticotypes during maintenance of these clones for up to 1,000 fissions indicated that corticotypes above 21 undergo a rapid downward shift, while corticotypes in the range of 18 to 21 change slowly. Although these observations are consistent with Nanney's earlier deduction of a "stability center" at corticotype 19, there appears to be little if any difference in the stability of perpetuation of corticotypes 18, 19 and 20. Within this "stability range," the inertia of maintenance of pre-existing corticotypes is sufficiently strong that sister clones derived from an exconjugant pair can remain different for 1,000 fissions. These findings are consistent with observations made earlier, and those in the present study, indicating that cells in stock cultures express a substantial range of corticotypes even when maintained with frequent transfer. The results suggest that mechanisms of spatially ordered structural assembly within the cell can show sufficient fidelity to allow long-term vegetative perpetuation of phenotypic differences without artificial selection.     

Baeocytes in the cyanobacterium <Emphasis Type="Italic">Pleurocapsa</Emphasis> sp.: Characterization of the differentiated cells produced by multiple fission

Electron microscopy of cyanobacterium Pleurocapsa sp. CALU 1126 revealed that multiple fission proceeds by successive binary fissions. The cultivation conditions were determined when the number of baeocytes (products of multiple fission) was comparable with that of macrocytes (products of binary fission), and cell sorting was achieved for the first time. Juvenile baeocytes were shown to differ from macrocytes in: (1) the absence of sheath; (2) the linear-peripheral configuration of their lamellar system; (3) lower content of phycobiliproteins and higher content of carotenoids; (4) the set of PSII polypeptides. Baeocytes can therefore be considered differentiated cells characterized by the uncoupling between energy and biosynthetic metabolism.     

Telomere sequence localization and karyotype evolution in higher plants

Data for chromosomal localization of theArabidopsis-type of telomeric sequence repeats (TTTAGGG)_n are compiled for 44 species belonging to 14 families of angiosperms, gymnosperms and bryophytes. For 23 species and seven families this is the first report. Species of all families, except theAlliaceae, revealed these sequences at their chromosome termini. This indicates thatArabidopsis-type telomeric repeats are highly conserved. It is inferred that they represent the basic telomere sequence of higher plant phyla. In theAlliaceae, a deviating sequence (and mechanism?) for the stabilization of chromosome termini has possibly evolved secondarily. Nine species revealed interstitial telomeric sequences in addition to the terminal ones, in three species (Vicia faba, Pinus elliottii, P. sylvestris) also at centromeric positions. Interstitial telomeric sequences may indicate karyotype reconstructions, in particular alterations of chromosome numbers by chromosome fusion — or inversions with one breakpoint within the terminal array of repeats. They may contribute to stabilization of chromosome breaks, especially centric fissions, and increase the frequency of meiotic and illegitimate recombination.     

Karyotype of the rare Johnston’s genet<Emphasis Type="Italic">Genetta johnstoni</Emphasis> (Viverridae) and a reassessment of chromosomal characterization among congeneric species

We present herein new data on karyotypes of members of the genusGenetta. G- and C-banded chromosomes of the Johnston’s genetGenetta johnstoni Pocock, 1908 (2n = 50 / FNa = 92) are described for the first time, and compared with those ofG. genetta (2n = 54 / FNa = 92). In addition, the standard karyotype ofG. maculata (2n = 52 / FNa = 96) was studied. A reassessment of taxonomic attribution of previously published material allowed us to characterize (2n, FNa, and chromosome morphology) the karyotypes of three genets, previously unknown (G. pardina, G. letabae andG. tigrina). Our results show that despite a rather low interspecific variability in 2n and FNa, all the species of genets (exceptG. pardina andG. maculata) appear differentiated when chromosomal morphology is taken into account. Although chromosomal banding data are limited, confrontation of G-band karyotypes with preliminary molecular phylogenetic results reveals that karyotypic evolution within the genusGenetta might involve various rearrangements like Robertsonian and tandem translocations, pericentric inversions, and centromere fissions; thus providing at least for some taxa a solid postzygotic isolation. Finally, our study suggests that cytogenetic analyses might constitute a useful tool for questioning interspecific boundaries, especially within the taxonomically debated complex of large-spotted genets.