CYTOGENETIC STUDIES IN CLARKIA,SECTION PRIMIGENIA. I. A CYTOLOGICAL SURVEY OF CLARKIA AMOENA

Snow , Richard . (U. California, Davis.) Cytogenetic studies in Clarkia, section Primigenia. I. A cytological survey of Clarkia amoena. Amer. Jour. Bot. 50(4): 337–348. Illus. 1963.—Clarkia amoena (2n = 14) shows great cytological diversity based on reciprocal translocations. About ½ the plants sampled were heterozygotes; in 2 of the subspecies the frequency was as high as ⅔. The metaphase configurations were varied, including rings of 4, 6, 8, 4 + 4, 6 + 4, and more unusual types forming a chain of 3 + a univalent or a chain of 4 + a univalent. The latter have only 13 chromosomes but are not true monosomies. Some plants with supernumerary chromosomes were found, in addition to a plant with only 12 chromosomes which was derived from a “monosomic” type. From crosses of various homozygous lines to a standard strain, it was found that the standard end arrangement of chromosomes occurs widely and commonly throughout the species. A minimum of 7 different end arrangements, giving a ring of 4 with the standard strain, and 1 giving a ring of 6, account for the chromosome pairing found in interstrain crosses. No doubt more would be found with further testing. Some of the translocations may be components of adaptive polymorphic systems, though there is no proof of this yet. No balanced lethal systems similar to those in the closely related genus Oenothera were found.     

Meiotic pairing and segregation of translocation quadrivalents in yeast

Meiotic pairing and segregation were studied in three different heterozygous reciprocal translocation strains of the baker’s yeast, Saccharomyces cerevisiae. Pachytene translocation quadrivalents were identified by a combination of immunofluorescence and fluorescence in situ hybridization and the karyotypes of meiotic products were determined by pulsed-field gel electrophoresis. The translocations differed with respect to the relative sizes of the chromosomes involved and the positions of translocation breakpoints, and produced translocation quadrivalents of widely different shapes. This allowed us to study the influence of the morphology of quadrivalents on their segregation behaviour. In all cases alternate predominated over adjacent segregation. 3:1 disjunction of chromosomes was more frequent when translocation breakpoints were close to the centromeres. If a translocation breakpoint was distant from the centromere, the occurrence of an intervening chiasma influenced the pattern of segregation. In general, quadrivalent formation and segregation resembled the behaviour of translocation heterozygotes in most higher eukaryotes. We therefore conclude that, although chromosome condensation does not occur in yeast metaphase, centromere orientation and chromosome disjunction are governed in a way similar to that of higher eukaryotes. Received: 6 February 1998; in revised form: 19 May 1998 / Accepted: 23 May 1998     

A naturally occurring chromosomal translocation in Myzus persicae (Sulzer)

Populations of M.persicae from glasshouses in Britain were found to have a structurally heterozygous chromosome complement. Biometric data from somatic metaphase chromosomes, and the pairing configurations of chromosomes at spermatocyte meiosis, indicate either a simple or a reciprocal translocation involving autosomes 1 and 3. All except one of the clones started from these populations showed high levels of esterase activity indicative of resistance to organophosphorus insecticides. The British translocated clones were all androcyclic in reproductive character, but of several distinct genotypes. A similar or identical translocation was found in seven out of nine clones started from M.persicae collected in the field in Japan. The translocation was present in Japanese M.persicae of all reproductive categories, being somehow maintained in the sexual breeding system of the aphid. In breeding experiments, translocation heterozygosity was inherited independently from colour and reproductive characters. The translocation was also found in a sample of M.persicae from southern California.     

Contribution of chromosomal imbalance to sperm selection and pre-implantation loss in translocation-heterozygous Chinese hamsters

Chinese hamster stocks with various structurally abnormal chromosomes have been produced by X irradiation. Among these stocks, 18 with various reciprocal translocations were used to investigate the participation of unbalanced gametes in fertilization and the development of unbalanced embryos. Among males as well as females heterozygous for the same translocation, there is no difference in the frequency of each disjunctional class. The participation of chromosomally unbalanced gametes in fertilization was investigated by chromosomal analysis of meiotic cells in heterozygotes for the 18 reciprocal translocations and pronuclei of fertilized ova obtained from crossing these heterozygotes. Compared with the expected frequencies from MII scoring, the frequencies of male pronuclei having a common deficiency of chromosome 1 (1q17-->1q42) or chromosome 3 (3p23-->3q31) decreased significantly in one-cell embryos. However, the frequencies of male pronuclei with other abnormalities were all consistent with those expected from MII scoring. In contrast, the frequencies of female pronuclei with any karyotype including the same abnormalities as those decreased in male pronuclei from the translocation heterozygotes were all consistent with those estimated from MII scoring. These results revealed clearly that most gametes with nullisomies as well as disomies for any chromosomal segments may participate in fertilization, whereas only male gametes nullisomic for certain segments of chromosomes 1 and 3 failed to participate in fertilization. The zygotic selection of chromosomal imbalance was also investigated by direct chromosomal and morphological analyses of preimplantation embryos from crosses between karyotypically normal females and male heterozygotes from the 18 stocks with various reciprocal translocations. These analyses revealed that some embryos were arrested in development at the two-cell stage. The karyotype of these two-cell embryos had a common deficiency in a segment of chromosome 1 or chromosome 2. Embryos with partial monosomy including chromosomes 1, 3, 4 and 5 showed arrested development at four- to eight-cell stages. Among day 4 embryos, some chromosomally unbalanced embryos, mainly with a deficiency of segments of chromosomes 1p, 1q, 2q, 5q, 7q and 8, had fewer blastomeres than karyotypically normal and balanced embryos. The homology between Chinese hamster and mouse chromosomes relating to abnormal embryogenesis at early stages has been partially confirmed from reported maps of chromosomes. The Chinese hamster is useful for further cytogenetic studies during the stages of meiosis and early embryogenesis.     

STABILITY OF A CHROMOSOMAL HYBRID ZONE IN THE CLARKIA NITENS AND CLARKIA SPECIOSA SSP. POLYANTHA COMPLEX (ONAGRACEAE)

Clarkia nitens and Clarkia speciosa polyantha (Onagraceae) are distinct chromosomal taxa differing by at least six reciprocal translocations. Where the taxa have come into contact, a chromosome boundary zone exists characterized by high levels of translocation heterozygosity due to at least 12 new chromosome arrangements which have evolved there. Previous studies have shown that these boundary arrangements are distributed such that they provide for full interfertility between adjacent populations. It was hypothesized that the geographic distributions of each of these arrangements will remain generally stable due largely to the very adverse effect that major changes would have on fertility. Evidence is presented here that over a ten year period the frequencies and geographic distributions of the chromosome arrangements within this boundary region have remained stable. The frequencies of the various chromosomal configurations (nine pairs, ring of four, two rings of four, ring of six, ring of four + ring of six, and ring of eight) from surveys in 1968 and 1978 have been analyzed statistically. In general, the analysis indicates that there have been no detectable changes over the 10-yr period.     

CYTOGENETIC STUDIES IN CLARKIA,SECTION PRIMIGENIA. II. A CYTOLOGICAL SURVEY OF CLARKIA ARCUATA AND CLARKIA LASSENENSIS

Clarkia arcuata and C. lassenensis are the 2 members of the subsection Flexicaules. Although closely related morphologically, they show very different patterns of chromosomal variability in nature. About 25% of the plants grown from wild seed of C. arcuata, a predominantly cross-pollinating species, were heterozygous for 1 or 2 translocations; such heterozygotes were found in 5 of the 9 populations sampled. An analysis of the chromosome pairing in intraspecific crosses indicated that at least 5 different translocations giving a ring of 4 with the “standard” strain, 2 giving a ring of 6, and 2 giving a ring of 8 are present in nature. No arrangement was found with widespread distribution, and it is impossible to say at present what might be the primitive arrangement of this species. One population was found to contain an inversion, a rearrangement which is very rare in Clarkia at the intraspecific level. In C. lassenensis, a predominantly self-pollinating species, only 6% (3 plants) of a sample of 53 were translocation heterozygotes, and these heterozygotes were found in only 2 of 13 populations. Intraspecific crosses indicated that one chromosome arrangement, the “standard,” was present throughout the species range.     

Synapsis and chiasma distribution in mice heterozygous for translocations in chromosomes 16 and 17

Electron microscopic analysis of synaptonemal complexes and analysis of chiasmata distribution in male mice heterozygous for Robertsonian translocation T(16; 17)7Bnr - (Rb7), for synaptonemal reciprocal translocation T(16;17)43H - (T43), in double heterozygotes for these translocations and in males with partial trisomy of the proximal region of chromosome 17 was carried out. Synaptic disturbances around the breakpoints of the translocations, such as asynapsis of homologous regions of partners and non-homologous synapsis of centromeric regions of acrocentric chromosomes, were revealed. Synaptic regularity in the proximal part of the chromosome 17 appeared to be affected by no t12 haplotype. Good coincidence between sizes of mitotic chromosomes and corresponding lateral elements of synaptonemal complexes was found for all chromosomes, with the exception of Rb7 in trisomics. In the latter karyotype, the proximal part of chromosome 17 involved in Robertsonian fusion seems to be shortened in the course of zygotene and never synapted with homologous segment of neither the acrocentric chromosome 17 nor large product of reciprocal translocation. Drastic increase in chiasmata frequency in the proximal part of chromosome 17 was revealed in heterozygotes for T43H and in trisomics, as compared with the double heterozygotes Rb7/T43. The latter finding was explained by the existence of two independent pairing segments in the former karyotypes.     

Mitomycin C-induced meiotic crossing-over on the interstitial segments in the Chinese hamster heterozygous for a reciprocal translocation

Using Chinese hamsters heterozygous for T(2;10)3Idr and T(1;3)8Idr reciprocal translocations, the authors studied mitomycin C (MMC)-induced crossing-over on the interstitial segments. Marker chromosomes with unequal-length chromatids resulting from crossing-over were clearly detectable, and the frequencies of such marker chromosomes were constant among individual males which were heterozygous for the same reciprocal translocation. The frequency of MMC-induced crossing-over on the interstitial segments increased roughly with increase in dose. These findings, therefore, indicated that marker chromosomes with unequal-length chromatids in translocation heterozygotes may be a useful indicator for detection of the cytogenetic effects of environmental mutagens on germ cells.     

Differentiated ZZ/ZW sex chromosomes in Apareiodon ibitiensis (Teleostei,Parodontidae): cytotaxonomy and biogeography

Conventional and molecular chromosomal analyses were carried out on three populations of Apareiodon ibitiensis sampled from the hydrographic basins of the São Francisco River and Upper Paraná River (Brazil). The results reveal a conserved diploid number (2n = 54 chromosomes), a karyotype formula consisting of 50 m‐sm + 4st and a ZZ/ZW sex chromosome system that has not been previously identified for the species. C‐banding analysis with propidium iodide staining revealed centromeric and terminal bands located in the chromosomes of the specimens from the three populations and allowed the identification of heteromorphism of heterochromatin regions in the Z and W chromosomes. The number of 18S sites located through fluorescent in situ hybridization (FISH) varied between the populations of the São Francisco and Upper Paraná Rivers. The location of 5S rDNA sites proved comparable in one pair of metacentric chromosomes. Thus, the present study proposes a ZZ/ZW sex chromosome system for A. ibitiensis among the Parodontidae, and a hypothesis is presented regarding possible W chromosome differentiation stages in this species through DNA accumulation, showing geographical variations for this characteristic, possibly as a consequence of geographical reproductive isolation.     

Comparative Cytogenetic Variation of the Salivary Gland Polytene Chromosomes in Chironomus riparius Mg., 1804 (Diptera, Chironomidae) from Two Polluted Biotopes of Bulgaria and Russia

The structural–functional variation ofChironomus riparius salivary gland polytene chromosomes was studied in two geographically isolated Palearctic regions, Bulgaria (village Pancharevo) and Russia (St. Petersburg). The two biotopes, where larvae were collected, were polluted with various heavy metals from anthropogenic sources. Hereditary paracentric heterozygous inversions were characteristic of the Russian population, whereas somatic paracentric or pericentric heterozygous inversions were more common in the Bulgarian one. All inversions occurred at low frequencies. Other aberrations found in the two populations included somatic deletions resulting in a pompon structure of chromosome IVG, heterozygous translocation between chromosomes IVG and IIIF, enlargement of individual disks, and the appearance of a heterozygous block close to the centromere of chromosome IVG. In addition, changes in functional activity of the nucleolus organizer and Balbiani rings (BRc/BRb) were observed. Several aberration breakpoints proved to coincide with satellites of the Alu and Hinf families.     

Is there an interchromosomal effect in reciprocal translocation carriers? Sperm FISH studies

Chromosome translocations have been known to affect disjunction of chromosomes unrelated to the translocation in the mouse and in Drosophila. However, in humans, an interchromosomal effect in chromosome translocations has not been demonstrated. The availability of techniques that allow the study of nondisjunction in sperm cells has permitted us to evaluate the possibility of an interchromosomal effect in male translocation heterozygotes. In this study, multicolor fluorescence in situ hybridization was used to determine levels of disomy for the clinically relevant chromosomes X, Y, 13, 18, and 21 in 332,858 spermatozoa from nine reciprocal translocation heterozygotes and nine controls with normal karyotypes. The specific translocations studied were as follows: t(10;12)(p26.1;p13.3), t(2;18)(p21;q11.2), t(3;19)(p25;q12), t(5;8)(q33;q13), t(11;22)(q23;q11), t(3;4)(p25;p16), t(8;9) (q24.2;q32), t(10;18)(q24.1;p11.2), and t(4;10)(q33;p12.2). Comparisons of disomy rates between carriers and controls were performed by using the Mann-Whitney test. Our results showed that the rates of sex chromosome hyperhaploidy were similar in controls (0.21%) and in translocation carriers (0.19%). Similarly, the frequencies of disomy for chromosomes 13, 18, and 21 did not differ significantly between controls and carriers (0.05% versus 0.08%, 0.07% versus 0.03%, and 0.14% versus 0.20%, respectively). Sex chromosome nondisjunction was more common than nondisjunction of chromosomes 13 and 18 both in controls (P=0.0057) and in carriers (P=0.0008). Similarly, the rates of chromosome disomy for chromosome 21 were higher than those for chromosomes 13 and 18 in both controls (P=0.0031) and translocation carriers (P=0.0057). In our study, the excess of chromosome 21 disomy versus disomy of the other autosomes was more pronounced in carriers than in controls. Thus, although the difference of disomy 21 between controls and carriers was not statistically significant, it is worthy of attention.     

Heterosis without selectional coadaptation inDrosophila ananassae

Summary The relative viabilities of homozygous and heterozygous karyotypes were measured by making crosses between strains ofD. ananassae homozygous for ST or inverted gene orders in the second and third chromosomes. The strains utilized during the present study originated from widely separated localities in India, Kuala Lumpur and Kota Kinabaru, Malaysia and Chian Mai, Thailand. The presence of heterosis in many interpopulation crosses is evident from the results which show that the inversion heterozygotes formed by chromosomes coming from distant populations exhibit heterosis. On the other hand, heterosis is absent in two intrapopulation crosses. Thus the present results provide evidence that heterozygosis for many genes and gene complexes does produce high fitness without previous selectional coadaptation.     

A reciprocal translocation radically reshapes sex‐linked inheritance in the common frog

X and Y chromosomes can diverge when rearrangements block recombination between them. Here we present the first genomic view of a reciprocal translocation that causes two physically unconnected pairs of chromosomes to be coinherited as sex chromosomes. In a population of the common frog (Rana temporaria), both pairs of X and Y chromosomes show extensive sequence differentiation, but not degeneration of the Y chromosomes. A new method based on gene trees shows both chromosomes are sex‐linked. Furthermore, the gene trees from the two Y chromosomes have identical topologies, showing they have been coinherited since the reciprocal translocation occurred. Reciprocal translocations can thus reshape sex linkage on a much greater scale compared with inversions, the type of rearrangement that is much better known in sex chromosome evolution, and they can greatly amplify the power of sexually antagonistic selection to drive genomic rearrangement. Two more populations show evidence of other rearrangements, suggesting that this species has unprecedented structural polymorphism in its sex chromosomes.     

Cytogenetic analysis of thioTEPA-induced translocations in the spermatids of male mice

Spermatocytes of 36 reciprocal translocation heterozygotes at diakinesis-MI stage were studied cytogenetically. There were 38.8 % cells with rings and 32.8 % - with chains and univalents among 3666 cells. It is concluded that intensive mutational process producing 30 % of translocation heterozygotes is characterized by random breakage along the chromosomes.     

Nondisjunction, disturbances in spindle structure, and characteristics of chromosome alignment in maturing oocytes of mice heterozygous for Robertsonian translocations

To correlate the chromosomal constitution of meiotic cells with possible disturbances in spindle function and the etiology of nondisjunction, we examined the spindle apparatus and chromosome behavior in maturing oocytes and analyzed the chromosomal constitution of metaphase II-arrested oocytes of CD/Cremona mice, which are heterozygous for a large number of Robertsonian translocation chromosomes (18 heterobrachial metacentrics in addition to two acrocentric chromosomes 19 and two X chromosomes). Spreading of oocytes during prometaphase 1 revealed that nearly all oocytes of the heterozygotes contained one large ring multivalent, apart from the bivalents of the two acrocentric chromosomes 19 and the X chromosomes, indicating that proper pairing and crossing-over between the homologous chromosome arms of all heterobrachial chromosomes took place during prophase. A large proportion of in vitro-matured oocytes arrested in metaphase II exhibited numerical chromosome aberrations (26.5% hyperploids, 40.8% hypoploids, and 6.1% diploids). In addition, some of the oocytes with euploid chromosome numbers (26.5% of the total examined) appeared to be nullisomic for one chromosome and disomic for another chromosome, so that aneuploidy levels may even be higher than expected on the basis of chromosome counts alone. Although oocytes of the complex heterozygous mice seemed able initially to form a bipolar spindle during first prometaphase, metaphase I spindles were frequently asymmetrical. Chromosomes in the multivalent did not align properly at the equator, centromeres of neighboring chromosomes in the multivalent remained maloriented, and pronounced lagging of chromosomes was observed at telophase I in oocytes obtained from the Robertsonian translocation heterozygotes. Therefore, disturbance in spindle structure and chromosome behavior appear to correlate with the chromosomal constitution in these oocytes and, ultimately, with failures in proper chromosome separation. In particular, reorientation appears to be a rare event, and malorientation of chromosomes may remain uncorrected throughout prometaphase, as we could not find many typical metaphase I stages in heterozygotes. This, in turn, could be the basis for malsegregation at anaphase and may ultimately induce a high rate of nondisjunction and aneuploidy in the oocytes of CD/Cremona mice, leading to total sterility in heterozygous females.     

MEGASPOROGENESIS AND GAMETOPHYTE SELECTION IN PAEONIA CALIFORNICA

Walters , James L. (U. California, Goleta.) Megasporogenesis and gametophyte selection in Paeonia californica. Amer. Jour. Bot. 49(7): 787–794. Illus. 1962.—In the ovules of Paeonia californica, a massive archesporium produces numerous (estimated at 30–40) megasporocytes, many of which complete meiosis. Several continue into gametophyte development, which is of the Polygonum type, and at the time of fertilization there are from 1 to 4 gametophytes per ovule. Rarely does more than 1 seedling per seed appear in germination. This species is characterized by extensive translocation heterozygosity, and other meiotic irregularities, in its natural populations. It shows a complete range from plants forming only pairs to those with all their chromosomes in rings at meiosis. The latter types have as high as 90% bad pollen. The course of events in the ovules is compared with the “Renner-effect” found in Oenothera. The multiple megasporocytes and subsequent events are seen as a mechanism which insures each ovule a high probability of containing a viable egg in spite of meiotic behavior which can produce 90% sterility, and thus insures high seed set in the translocation heterozygotes.     

Translocations of Chromosome End-Segments and Facultative Heterochromatin Promote Meiotic Ring Formation in Evening Primroses

Due to reciprocal chromosomal translocations, many species of Oenothera (evening primrose) form permanent multichromosomal meiotic rings. However, regular bivalent pairing is also observed. Chiasmata are restricted to chromosomal ends, which makes homologous recombination virtually undetectable. Genetic diversity is achieved by changing linkage relations of chromosomes in rings and bivalents via hybridization and reciprocal translocations. Although the structural prerequisite for this system is enigmatic, whole-arm translocations are widely assumed to be the mechanistic driving force. We demonstrate that this prerequisite is genome compartmentation into two epigenetically defined chromatin fractions. The first one facultatively condenses in cycling cells into chromocenters negative both for histone H3 dimethylated at lysine 4 and for C-banding, and forms huge condensed middle chromosome regions on prophase chromosomes. Remarkably, it decondenses in differentiating cells. The second fraction is euchromatin confined to distal chromosome segments, positive for histone H3 lysine 4 dimethylation and for histone H3 lysine 27 trimethylation. The end-segments are deprived of canonical telomeres but capped with constitutive heterochromatin. This genomic organization promotes translocation breakpoints between the two chromatin fractions, thus facilitating exchanges of end-segments. We challenge the whole-arm translocation hypothesis by demonstrating why reciprocal translocations of chromosomal end-segments should strongly promote meiotic rings and evolution toward permanent translocation heterozygosity. Reshuffled end-segments, each possessing a major crossover hot spot, can furthermore explain meiotic compatibility between genomes with different translocation histories.     

Chromosomes tell half of the story: the correlation between karyotype rearrangements and genetic diversity in sedges,a group with holocentric chromosomes

Chromosome rearrangements may affect the rate and patterns of gene flow within species, through reduced fitness of structural heterozygotes or by reducing recombination rates in rearranged areas of the genome. While the effects of chromosome rearrangements on gene flow have been studied in a wide range of organisms with monocentric chromosomes, the effects of rearrangements in holocentric chromosomes—chromosomes in which centromeric activity is distributed along the length of the chromosome—have not. We collected chromosome number and molecular genetic data in Carex scoparia, an eastern North American plant species with holocentric chromosomes and highly variable karyotype (2n = 56–70). There are no deep genetic breaks within C. scoparia that would suggest cryptic species differentiation. However, genetic distance between individuals is positively correlated with chromosome number difference and geographic distance. A positive correlation is also found between chromosome number and genetic distance in the western North American C. pachystachya (2n = 74–81). These findings suggest that geographic distance and the number of karyotype rearrangements separating populations affect the rate of gene flow between those populations. This is the first study to quantify the effects of holocentric chromosome rearrangements on the partitioning of intraspecific genetic variance.     

Strukturheterozygotie beiVicia faba

Summary The karyotypes and the meiotic behaviour of two spontaneous reciprocal translocations and one pericentric inversion are described. One of these translocations was characterized by chiasma-formation in the interstitial-segments, the other was not. The types of orientation and distribution of the chromosomes from the chain-configurations in meta-anaphase I are specified, compared mutually and with those of translocation rings and chains in maize and the similarities and differences are pointed out. In the range of the heterozygous pericentric inversion there was no pairing in meiotic prophase and therefore no chiasma formation and sterility. It will be tried to cross especially one of the translocations with the inversion-type in order to rebuild the karyotype ofVicia faba. In this case it would be possible to distinguish between all five pairs of small chromosomes according to their morphological structure, a possibility very important in localizing chromosome aberrations on the chromosomes ofVicia faba.

---

---

Mit 15 Abbildungen     

Estimation of Fitness Components in DROSOPHILA MELANOGASTER. II. Influence of Reduced Transmission Frequency

Results are presented of further analyses of the significant effects of reciprocal crosses reported by Katz and Cardellino (1978) in regard to viability indices of wild-type second chromosome heterozygotes. The observed differences between reciprocal crosses can be explained by the existence of reduced transmission frequencies of the wild-type homologue from Pm/+ and Cy/+ paternal parents. Mean estimates of transmission frequencies from Pm/+ and Cy/+ males in California and Japan populations are significantly less than the Mendelian expectation of 1/2. The transmission frequencies of +(i) chromosomes from Pm/+(i) and Cy/+(i) males are also found to be positively correlated in the California and pooled populations, suggesting that the degree of distortion is primarily due to the +(i) chromosome rather than to Cy or Pm. A sufficient estimator of relative viability that is independent of distorted transmission frequencies is derived for use in the Cy/Pm technique of viability estimation.