The supernumerary segment system of Stethophyma

Three species of the genus Stethophyma have been cytologically examined and all three show variation both for supernumerary heterochromatic segments and for the distribution of standard heterochromatin among the autosomes. The European species, S. grossum, for example, shows considerable interpopulation variation for standard heterochromatin while two of the populations, from Spain and Austria, show supernumerary segment polymorphism. The segments are located interstitially on the S₁₁ chromosome but occupy different positions in the different populations. — In all species, the presence of the extra heterochromatic segments increases the mean chiasma frequency. Moreover, the influence of the segments upon mean chiasma frequency is different in different populations and in different species. In the Spanish population, the increase is both intra- and interchromosomal whereas in Austria the influence of the segment is completely interchromosomal. — In the American species, S. gracile and S. lineatum, where supernumerary heterochromatic segments are carried on both S₁₀ and S₁₁ chromosomes, the effect on chiasma frequency shows a dosage relationship, an increase in the number of segments per individual being correlated with an increase in mean chiasma frequency. It is suggested that the interstitial segments found in all species have originated by direct duplication of chromosome material. By contrast the terminal segments in S. lineatum and S. gracile may be derived by translocation from a B-chromosome since such a chromosome has been found in one individual of the former species. — The variation in segment structure and the distribution of standard heterochromatin, among the European species of S. grossum suggests that these systems have evolved independently in different populations.On educational leave from the Forest Research Laboratory, Fredericton, N. B. Canada.     

Spontaneous chromosome mutations in Truxaline grasshoppers

Three distinct mutant conditions are described in single male individuals from three species of short horn grasshopper. Of these, one is an entire germ line mutant of Myrmeleotettix maculatus, heterozygous for a centric fusion between single M₄ and M₅ telocentric chromosomes. In contrast, the remaining two mutants are present in mosaic form. One is heterozygous for an L₁-M₄ interchange in Omocestus viridulus, the other tetrasomic for the M₄ chromosome in Chorthippus parallelus which in addition is characterised by the inclusion of a supernumerary heterochromatic segment on one S₈ homologue. Centric fusion in Myrmehotettix maculatus has neither disturbed the chiasma potential of the elements constituting the fusion multiple nor, has it apparently influenced the production of balanced gametes. The pattern of chiasma formation in the L₁-M₄ interchange multiple lends support to the contention that the process of chiasma formation originates near the distal end of chromosome arms in Omocestus viridulus. There is no interaction between the two mutant conditions of tetrasomy and the presence of supernumerary segments in Chorthippus parallelus. Moreover, because of the precocious nature of two of the four M₄ homologues there is little tendency to form multivalents. The two M₄ bivalents share a similar mean chiasma frequency.     

Supernumerary segments in five species of grasshoppers (Orthoptera: Acridoidea)

Supernumerary segments have been observed in five species of grasshoppers: Calliptamus barbarus, Oedipoda fuscocincta, Acrotylus insubricus, Omocestus raymondi and Chorthippus ariasi. In four cases they are located in the S-chromosomes, but in A. insubricus they are carried by the megameric pair (M₉). In C. barbarus and O. fuscocincta we have observed non-homologous association during diplotene between the extra segments and the X-chromosome. The supernumerary segments of these two species are distally located. However, in anaphase-I the unequal bivalents divide reductionally in 30% and 20% of the cells, respectively. Finally, the supernumerary segments of C. barbarus do not produce any effect on mean chiasma frequency, but they decrease significantly the between-cell variance of chiasmata in males carrying them.     

Male and female segregation distortion for heterochromatic supernumerary segments on the S8 chromosome of the grasshopper Chorthippus jacobsi

The mode of inheritance of supernumerary segments located on three different chromosome pairs was investigated in controlled crosses with specimens of the grasshopper Chorthippus jacobsi. While extra segments located on chromosomes M₅ and M₆ showed Mendelian inheritance, that on S₈ did not. Thus, the two supernumerary heterochromatic chromosome segments located distally on the S₈ chromosome accumulated through non-Mendelian transmission through both sexes. The observed transmission patterns may be explained by gametic selection for spermatozoa carrying segmented S₈ chromosomes, in addition to meiotic drive for segmented S₈ chromosomes in heterozygous females. The significance of these findings for the maintenance of these polymorphisms in natural populations is discussed.by S.A. Gerbi     

Parallel polymorphism for supernumerary segments inChorthippus parallelus (zetterstedt)

A complex and parallel pattern of polymorphism for heterochromatic supernumerary segments in the M₇ and S₈ chromosomes has been found in 14 populations of the meadow grasshopperChorthippus parallelus. Nine distinct karyotype classes for these two chromosome pairs occur though they are not equally represented in different populations. Populations differ also with respect to the frequency of supernumerary segments they contain. In all populations the presence of supernumerary segments leads to a significant elevation of mean cell chiasma frequency compared to individuals from the same population lacking such segments. The extent of the effect appears to differ in different populations. The observed frequencies of S₈ karyotypes conform to the expectations of a Hardy-Weinberg distribution. Those of the M₇, however, do not, and in all but one of the 14 populations there is a significant excess of homokaryotypes. In the Ashurst population 26% of the individuals sampled were characterised by germ-line polysomy for the M₄ chromosome, either in the form of entire tetrasomics or more usually as mosaics ranging from tri- to hepta-somics. In all these polysomics the M₄ chromosomes in excess of two were regularly heteropycnotic at first meiotic prophase from zygotene to diakinesis. As a consequence of this multivalents are rare. Extra M₄ chromosomes do not modify the chiasma characteristics of the other chromosomes in the complement. Nor do they modify the action of the supernumerary segments in any way.     

The effect of supernumerary chromosomes on meiosis in Puschkinia libanotica (Liliaceae)

The presence of the partly heterochromatic supernumerary chromosomes in pollen mother cells of Puschkinia libanotica raises the chiasma frequency in the A chromosome bivalents. The pattern of chiasma distribution along each of the five A bivalents was related to the DNA labelling pattern of mitotic chromosomes. Regions that showed heavy labelling at the end of the DNA synthetic phase had fewer chiasmata than lightly labelled regions. As this relation is the opposite to that found by Rees and Evans in another species we regard any correlation between labelling pattern and chiasma distribution as fortuitous.     

A polymorphism for heterochromatic supernumerary segments in Chorthippus parallelus

Supernumerary heterochromatic segments have been found in the S₈ bivalent of two populations of Chorthippus parallelus. These segments occur both in the heterozygous and the homozygous states. In both populations the frequencies of basic homozygotes, structural heterozygotes and structural homozygotes conform to a Hardy-Weinberg distribution. In the structurally heterozygous and homozygous states there is a significant increase in the chiasma frequency of the bivalents other than the S₈ itself. It is suggested that the adaptive role of these supernumerary segments can be explained in terms of this interchromosomal effect.To Professor J. Seiler in commemoration both of his 80th birthday and his many distinguished years of cytological service.     

Chiasma redistribution in the presence of different sized supernumerary segments in a grasshopper: dependence on nonhomologous synapsis.

Eight different sized supernumerary segments located at distal ends of the long arms of chromosomes M4, M5, M6, and S8 of the grasshopper Stenobothrus festivus were studied in males with regard to the synaptic process and chiasma distribution in the bivalents that carry them. The M4, M5, and M6 bivalents heterozygous for extra segments were always monochiasmate, in contrast to their bichiasmate condition observed in basic homozygotes. Furthermore, the presence of any of these extra segments led to chiasma redistribution in the carrier bivalents, so that such chiasmata were formed preferentially further away from the extra segment. The intensity of this effect is dependent on the size of the segment. Not all heteromorphic bivalents exhibited synaptonemal complexes with equalized axes at pachytene, but there was always a variable proportion of heterosynapsis around the distal ends of the long arms that was dependent on both the size of the segment and the size of the carrier chromosome. It is proposed that the absence of chiasmata in nonhomologous synapsed regions is responsible for the results obtained. Length measurements of the different extra segments and their carrier chromosomes between pachytene and diplotene indicated that synaptonemal complex is underrepresented in supernumerary heterochromatin.     

Genetic and environmental components of chiasma control

Variation in chiasma frequency within and between individuals has been investigated in Schistocerca gregaria and Stethophyma grossum. By taking sequential samples of the same testis in S. gregaria it has been demonstrated that there is considerable variation in chiasma frequency between times, within individuals, and that this variation is not evenly distributed between individuals of the same population. The response and recovery patterns after exposure to successive temperature treatments also indicate a differential sensitivity of individuals within the experimental population. — An analysis of interfollicular variation in chiasma frequency has revealed significant differences between follicles within individuals both in S. gregaria and a Spanish population of S. grossum. A comparable analysis on individuals of S. grossum carrying supernumerary segments shows that the presence of such segments increases the amount of variation between follicles within individuals and also between individuals within the population. — The distribution of cell chiasma frequencies in S. gregaria is normal whereas in Stethophyma cell frequencies approximate to a Poisson distribution. The structurally different supernumerary segments, present in Austrian and Spanish populations of S. grossum, both increase mean chiasma frequency and they modify the between cell variance in different ways — the former interchromosomally and the latter intrachromosomally. — The differences in chiasma frequency between follicles and between times within individuals and the differential reaction to heat shock reflect differences in the pattern of genotype-environment interaction. Similarly it can be argued that the presence of polymorphisms involving supernumerary material may play an equivalent role by increasing the between cell or between bivalent variance within individuals, a unique form of genotype-environment interaction at the endophenotypic level. — Such variation in genotype sensitivity to environmental modification has important adaptive value especially in organisms which are subject to recurrent, often random, changes in their environment.     

Chromosomal similarities and differences among four Neotropical Elateridae (Conoderini and Pyrophorini) and other related species, with comments on the NOR patterns in Coleoptera

This work deals with the comparative cytogenetic analysis of four Neotropical Elateridae species and reviews the nucleolar organizer region (NOR) patterns on Coleoptera chromosomes, for the first time. The cytogenetic characterization of Conoderus malleatus (Conoderini), Pyrearinus candelarius, Pyrophorus divergens and Pyrophorus punctatissimus (Pyrophorini) was accomplished through the study of mitotic and meiotic cells submitted to standard (Giemsa) and differential staining [silver impregnation and GC‐specific chromomycin A₃ (CMA₃) plus AT‐specific 4′‐6‐diamidino‐2‐phenylindole (DAPI) fluorochromes]. The analysis of spermatogonial cells revealed the diploid numbers: 2n = 17 in C. malleatus and 2n = 15 in P. candelarius, P. divergens and P. punctatissimus. In these species, the X0 type sex‐determination system and the acrocentric morphology of almost all chromosomes were observed. The study of meiotic cells of the four species revealed the occurrence of total synapsis between the autosomes, the presence of one terminal or interstitial chiasma in the majority of the bivalents, and the reductional behaviour and regular segregation of all chromosomes. Although the three Pyrophorini species demonstrated many similar karyotypical characteristics, there was one discrepancy, which was noted in the diplotene cells and concerns the number of bivalents with two chiasmata; P. candelarius only presented one bivalent, P. divergens showed two bivalents and P. punctatissimus exhibited up to four bivalents with two chiasmata. Testicular cells impregnated with silver nitrate demonstrated two terminal NORs located on the fourth autosomal pair of the Conoderini species and on the second autosomal pair of the three Pyrophorini representatives. Use of CMA₃/distamycin A (DA)/DAPI staining on the P. candelarius and P. punctatissimus chromosomes revealed that the CMA₃ labelled regions were coincident with the NORs. The main strategies of karyotypical differentiation that have occurred among the four Elateridae species and other related species, and the general trends of the NOR shifts during Coleoptera chromosomal evolution are discussed in this work.     

SUPERNUMERARY CHROMOSOMES IN TRADESCANTIA II. EFFECT OF COLCHICINE ON MEIOTIC ISOCHROMOSOME CONFIGURATIONS

The effect of colchicine on meiotic pairing and configuration frequencies of three homologous isosupernumerary chromosomes was investigated. In the absence of colchicine, the three isochromosomes displayed a high degree of interchromosomal pairing and chiasma formation. As a consequence, a high frequency of bivalents and trivalents were observed at diakinesis-metaphase I. The unique structure of isochromosomes enables them to pair intrachromosomally (i.e., foldback pairing) yet the preferential occurrence of interchromosomal pairing suggests that all six arms of the three isosupernumerary chromosomes were in close association prior to or upon initiation of synapsis. Supernumerary chromosomes in microsporocytes treated during presynapsis or early synapsis with colchicine exhibited a significant reduction (P < 0.001) in the number of bivalents and trivalents at diakinesis. However, there was no reduction in overall chiasma frequency among supernumeraries due to the induction of increased intrachromosomal pairing and chiasma formation. A colchicine-sensitive association or alignment of homologues preceding effective pairing has been demonstrated in standard chromosomes of a number of plant species. This study provides the first evidence to indicate that at least certain supernumerary chromosomes may display presynaptic association as well. The results also support the strongly held contention that colchicine is not directly preventing or inhibiting the actual formation of chiasmata, since no reduction in chiasma frequency was observed in the isochromosomes.     

Chiasma frequency,distribution and interference maps of mouse autosomes

Chiasma frequencies were analysed and chiasma positions measured in diakinesis/metaphase I autosomal bivalents from oocytes and spermatocytes of F₁ hybrid C3H/HeH×101/H mice. Twenty chromosome size ranks, including the presumptive X bivalent, could be distinguished in oocytes, and nineteen autosomal ranks plus the XY pair spermatocytes. Overall, mean cell chiasma frequencies of the two sexes did not differ significantly once the contribution of the presumptive X bivalent and the XY pair were taken into account. Sex related differences in chiasma distribution patterns were evident, however. In monochiasmate bivalents, the chiasma was most commonly located interstitially in oocytes while in spermatocytes it could be either interstitial or distal. In dichiasmate bivalents, the chiasmata tended to be more centrally located in oocytes than in spermatocytes. Minimum inter-chiasma distances did not appear to show any great variation in chromosome pairs of different sizes, however, mean inter-chiasma distances did increase with the bivalent length. The minimum-inter chiasma distance data suggest that chiasma interference is complete over a chromosomal segment equating to approximately 60 Mb. Measurement of the positions of chiasmata along chromosome arms open up the possibility of producing chiasma-based genetic maps for all the autosomes of the mouse.     

Parallel polymorphism for supernumerary segments in Chorthippus parallelus (Zetterstedt)

Heterochromatic supernumerary segments, present in either or both the M₇ and the S₈ chromosomes of the Ashurst (AS) population of Chorthippus parallelus, lead to a significant elevation in mean cell chiasma frequency. This effect appears to be dosage independent within both the S₈ and the M₇ karyomorphs when these are considered separately. Neither is there any marked interaction between the two systems when they are combined with one another. The selective factors controlling the distribution of S₈ karyomorphs within the population appear, however, to act quite independently of those which regulate the M₇ karyomorph pattern. This is revealed in two ways. First, whereas the frequency of S₈ karyomorphs conform to the expectations of a Hardy-Weinberg distribution those of the M₇ do not. Secondly, though the pattern of distribution of S₈ types is uniform throughout the AS population that of the M₇ is not. This heterogeneity of M₇ karyomorphs implies that the population is in some way internally stratified in terms of its breeding structure.     

Supernumerary chromosomes in Tradescantia I. Meiotic behavior of three homologous isochromosomes

The meiotic behavior of three homologous iso-supernumerary chromosomes in an individual of Tradescantia ohiensis Raf. has revealed sevealed several interesting features. There is a high frequency of pairing-partner exchange within bivalents and trivalents. The mean chiasma frequency per chromosome is relatively high (1.046) for supernumerary chromosomes and is not significantly different from that of the much larger normal (A) chromosomes. The high frequency of trivalent and bivalent configurations would seem to suggest that the isosupernumerary chromosomes are premeiotically dispositioned to allow for full random pairing among all six homologous arms.     

Heterosynapsis and suppression of chiasmata within heterozygous pericentric inversions of the Sitka deer mouse

The patterns of chromosomal pairing and chiasma distribution were analyzed in male Sitka deer mice (Peromyscus sitkensis) polymorphic for terminally positioned pericentric inversions of chromosomes 6 and 7. Gand C-banding of somatic metaphases indicated that the inversions involved 30% and 40% of chromosomes 6 and 7, respectively. Analysis of silver-stained synaptonemal complexes in surface-spread zygotene and pachytene nuclei from heterozygous individuals revealed that inversion loops were not formed. The inverted segments proceeded directly to heterosynapsis without an intervening homosynaptic phase, and the heteromorphic bivalents remained straight-paired throughout pachynema. C-banded pachytene nuclei corroborated the occurrence of heterosynapsis, as the heteromorphic bivalents exhibited nonaligned centromeres. Analysis of diplonema and diakinesis indicated that crossing over had not occurred within the heterosynapsed inverted segments. The observation of chiasma suppression within the inversions indicates that pericentric inversion heterozygosity does not lead to the production of unbalanced gametes. Heterosynapsis of the inverted segments during zygonema and pachynema and the resulting chiasma suppression therefore represent a meiotic mechanism for the maintenance of pericentric inversion polymorphisms in this population of P. sitkensis.     

Heterochromatin content and chiasma distribution in the megameric chromosomes of Stethophyma gracile and S. lineatum (Orthoptera: Acrididae)

Single populations of Stethophyma gracile and S. lineatum from Eastern Canada show substantial differences in the relative amount of heterochromatic and euchromatic material in the extensively heterochromatic (megameric) M₉ autosome. No differences in chromosome phenotype between these two species have been previously reported. A new polymorphism with respect to an interstitial, supernumerary H-block was also found in the S. lineatum population, and it appears to have some interchromosomal influence upon chiasma condition. These findings suggest that the North American Stethophyma forms may parallel the extensive and deme-specific heterochromatin variation found in the European populations of the Palaearctic S. grossum. Special attention was focused upon the structural organization of the proximal and distal H-segments on the M₉ during the early prophase of the first meiotic division in the male sex. These conspicuous, hetregions, prior to diplotene, are clearly interrupted and contain short, euchromatic sequences. This condition can readily account for the absence of chiasma localization and the distribution of chiasmata along the H-blocks at diplotene-diakinesis in the megameric element. This is the only member of the complement showing no evidence of pronounced proximal or proximal/distal chiasma localization at diplotene. Possible explanations for this interesting phenomenon are suggested.     

Comparative chiasma analysis using a computerised optical digitiser

A new computerised technique has been devised for measuring the distribution of chiasmata along diplotene bivalents. The method involves the introduction into the field of view of the microscope, of a fine light spot which can be accurately manipulated along the chromosomes of each bivalent. The data recorded include (a) the positions of the chiasmata along the bivalent in terms of their relative distances from the centromere and (b) the individual bivalent and cellular chiasma frequencies. — The method has been applied to the analysis of chiasma distribution patterns in the two known species of the genus Caledia, C. species nova 1 and C. captiva and in two chromosomal races of the latter. Statistical tests indicate that within bivalents at least 40% of the comparative distribution patterns of chiasmata between races and species are significantly different. Similar comparisons between populations within races reveal only 18% significant differences. — The observed distribution patterns of chiasmata in this genus suggest that chiasma formation is sequential from centromere to telomere. — The variation in the frequency and distribution of chiasmata between races and species suggests that the interference distances between successive chiasmata are, at least partially, independent of chiasma frequency and position. — The interracial and interspecific differences in chromosome structure are correlated with changes in chiasma pattern.     

SUPERNUMERARY CHROMOSOMES IN SAXIFRAGA VIRGINIENSIS (SAXIFRAGACEAE)

Acetocarmine squashes of root tips have demonstrated that 2n = 20 and 38 in Saxifraga virginiensis. These contrast with the earlier reported count of 2n = 28 for this species. In several populations supernumerary chromosomes were detected. Both intrapopulational and interpopulational variation in supernumerary chromosome number were detected, with the largest number of supernumerary chromosomes observed being six. Because these supernumerary chromosomes are equal in size to many of the smaller A chromosomes during mitotic metaphase, the presence of supernumerary chromosomes in this species could not be ascertained by analysis of mitotic metaphase preparations alone. During mitotic prophase, however, the supernumerary chromosomes of S. virginiensis are highly heterochromatic, appearing more densely coiled and darkly stained than the A chromosomes. This characteristic facilitated the recognition of supernumerary chromosomes in this species. The similarity in size of A and supernumerary chromosomes during mitotic metaphase and the observation of six supernumerary chromosomes in one population suggest that the count of 2n = 28 reported earlier for S. virginiensis may actually be a misinterpretation of 2n = 20 plus 8 supernumerary chromosomes. Furthermore, these findings and the observation of this same supernumerary chromosome phenomenon in other species of Saxifraga raise the possibility that some of the many disparate chromosome counts attributed to aneuploidy in the large genus Saxifraga may also be the result of misinterpretations of supernumerary chromosomes as A chromosomes.     

THE GENUS COLLINSIA. XVI. SUPERNUMERARY CHROMOSOMES

Dhillon , T. S., and E. D. Garber . (U. Chicago, Chicago, Ill.) The genus Collinsia. XVI. Supernumerary chromosomes. Amer. Jour. Bot. 49 (2) : 168–170. 1962.—The pairing and transmission of supernumerary chromosomes in Collinsia solitaria and in hybrids between C. sparsiflora subsp. arvensis and C. bruceae were studied. Two supernumerary chromosomes usually formed a bivalent with 1 chiasma at metaphase I; 4 supernumerary chromosomes occasionally yielded a trivalent and univalent at this stage. Plants with 2–4 supernumerary chromosomes were fertile and plants with 5–8, sterile. Plants with a given number of supernumerary chromosomes when used as seed or pollen parent gave gametes with a higher number of such chromosomes.     

Polymorphism involving heterochromatic segments in Metrioptera brachyptera

A complex pattern of polymorphism involving terminal heterochromatic segments on L₃ and L₄ chromosomes has been uncovered in eight populations of Metrioptera brachyptera. There are individuals in every population which carry reduced segments on one or both L₄'s. In six populations, enlarged heterochromatic segments have been encountered on the L₃ chromosomes in some individuals. The L₄ system is almost certainly stable although the frequency of L₄ karyotypes does not conform to a Hardy-Weinberg distribution in all populations. Stability of the L₃ polymorphism could not be ascertained. A reduction of L₄ heterochromatin leads to a significant rise both in mean cell chiasma frequency and between cell variance. The effect on chiasma frequency is transchromosomal. The normal pattern of strict chiasma localisation tends to be disrupted in germ lines which include modified L₄ chromosomes. There is a reduction in the number of proximal and distal chiasmata and an increase in the frequency of interstitial ones. It is proposed that the standard L₄ heterochromatin may function in conserving heterozygosity and promoting uniformity between parent and offspring. Partial removal may lead to an effective increase in recombination and produce a greater diversity of genotypes for selection to act upon.