Cytogenetics of the parthenogenetic grasshopper Warramaba (formerly Moraba) virgo and its bisexual relatives

Hybrids of both sexes were obtained from the reciprocal crosses, carried out in the laboratory, between the species believed to have given rise, by hybridization some thousands of years ago, to the parthenogenetic species W. virgo. All males with a Y-chromosome derived from P196 died before reaching the adult stage, but two males from the reciprocal cross (i.e., with a Y from P169) survived up to the adult stage. Their testes were small but normal in structure and histology. At the first meiotic division almost all the chromosomes were univalents, 0–2 bivalents being formed. Some chiasmata between non-homologous chromosomes were present. Segregation at first anaphase is irregular so that sperms with 3–15 chromosomes are formed. Such hybrids appear to be entirely sterile.     

Cytogenetics of the parthenogenetic grasshopper Warramaba (formerly Moraba) virgo and its bisexual relatives

The karyotypes of Warramaba virgo and the two bisexual species which gave rise to it, by hybridization, have been studied by the C- and G-banding techniques. W. virgo consists of a number of clones which differ by a limited number of inversions, translocations and fusions and also in C-banding pattern. In general, each local population consists of a single clone, although instances are known where two clones exist at the same locality. A total of fifteen cytological clones are here recognised and described. Some of these are known from single localities, while others have been found at several collecting sites in the same general area. The differences between them in banding pattern are relatively minor, i.e. the architecture of the karyotype has been highly conserved. A total of 50 C-bands have been recognised in W. virgo, but some of these are only present in a single clone and most clones have 39–40 bands. Most of these are centromeric or interstitial; a few are terminal (telomeric) in the short arms. There are three types of bands, dark-staining centromeric ones, intermediate ones that are somewhat less densely staining, and lighter ones that appear grey even after successful C-banding. The C-banding pattern of W. virgo provides additional evidence for its origin by hybridization between the bisexual species P196 and P169. It is possible that W. virgo arose only once (i.e. from one or more matings between P196 and P169 individuals in the same deme, within a few generations). However, two clones (Zanthus and Boulder), which are themselves closely similar, differ so much in banding pattern from all others that they may have arisen independently.     

Cytogenetics of the parthenogenetic grasshopper Warramaba (formerly Moraba) virgo and its bisexual relatives

The distribution of late-replicating segments along the chromosomes of five clones of W. virgo is described. Some, but not all of these segments correspond to C-bands. In general, the autoradiographic profiles (histograms of linear grain density along the length of chromosomes labeled with tritiated thymidine in late S-phase) show strong resemblances throughout the five clones. However, some significant differences exist, and these are particularly marked in the case of the Boulder clone, which is anomalous in many other respects. — A similar study has also been carried out on the two bisexual species of Warramaba (P169 and P196) that gave rise, by hybridization more than half a million years ago, to the parthenogenetic W. virgo. In the case of P169, the autoradiographic profiles of the three large chromosomes (X+A, B+5, CD) which it has contributed to the W. virgo karyotype are extremely similar to those of the corresponding chromosomes in the virgo clones we have studied. In the case of P196 there is likewise, in most instances, a close resemblance of the autoradiographic profiles of the AB, X₁ and CD chromosomes to those of the same chromosomes in the virgo clones, but that of the X₁ shows no particular resemblance to the anomalous profile of the X₁ in the Boulder clone, in which the X₁ has undergone a structural reorganisation. The autoradiographic profile of the P196 CD chromosome does, however, show a much closer resemblance to that of the corresponding chromosome in the Boulder clone than to those of the CD₁₉₆ in the other four virgo clones studied. These investigations confirm the considerable evolutionary stability of DNA replication patterns.     

Cytogenetics of the parthenogenetic grasshopper Moraba virgo and its bisexual relatives

An undescribed bisexual grasshopper species closely related to the all-female parthenogenetic Moraba virgo has an X₁X₂Y sex chromosome mechanism which incorporates 7 chromosomes of the ancestral karyotype (the original X and three pairs of autosomes). Apparently three separate chromosomal fusions have occurred, one of these being a tandem fusion followed by a crossover which stabilized it in the sex chromosome system. M. virgo probably arose from an ancestor which had X₁X₂Y males but lacked the tandem fusion present in the new species, the females of virgo having the constitution X₁X₁X₂O.Supported by Public Health Service Grant GM-07212 from the Division of General Medical Sciences, U.S. National Institutes of Health, and by a grant from the Australian Research Grants Committee.     

Cytogenetics of the parthenogenetic grasshopper Warramaba virgo and its bisexual relatives

Combinations of DNA-binding fluorescent dyes and counterstains that enhance selectivity and contrast in primary stain fluorescence were used to differentiate types of C-bands in the genus Warramaba. Chromomycin A₃ (in conjunction with two A-T binding counterstains), which identifies chromosome segments enriched in G-C base pair clusters, stains only a minority of the C-bands in Warramaba species, but these include all those known to contain 18S + 26S rRNA cistrons and most of those containing 5S rRNA genes. DAPI/actinomycin D fluorescent staining is positive for a very few bands, including two (in the Standard phylad of W. virgo) that are at or adjacent to sites containing 5S rRNA cistrons. One of the latter regions is also positively stained by DAPI/distamycin A which, in addition, highlights some centromeric bands. The fluorescent staining patterns of the Standard and Boulder-Zanthus phylads of W. virgo are significantly different, confirming their independent origin by hybridization between different races of the ancestral species “P169” and “P196”.     

Cytogenetics of the parthenogenetic grasshopper Warramaba virgo and its bisexual relatives

A study of the chromosomal location and genomic organization of the ribosomal RNA cistrons in the genus Warramaba, involving in situ hybridization and restriction enzyme analysis as well as C- and N-banding and silver staining, has confirmed that the parthenogenetic species W. virgo has two phylads. These phylads appear to have originated independently by hybridization between the precursors of the present day bisexual species P169 and P196. The clones of the Standard phylad of W. virgo have their 18S+26S rDNA cistrons located in C-bands 4, 44 and 49, while those of the Boulder-Zanthus phylad have them in C-bands 50, 74 and 87.5. The relative numbers of the ribosomal genes at the different sites vary greatly from clone to clone and are closely correlated with the width of the corresponding C- and N-bands. Site 49 of the ribosomal cistrons is present as a separate band in the eastern race A of P196 but has been incorporated into band 50 in the western race B of this species. The former race is assumed to be ancestral to the Standard phylad of W. virgo, the latter to the Boulder-Zanthus phylad, but there has been loss of the 74 and 87.5 sites in the the Standard phylad and the 4 and 44 sites in the Boulder-Zanthus clones. The ribosomal cistrons in W. picta, a species with a primitive karyotype, occur in several sites, only some of which have counterparts in P169 and P196. The 5S rDNA cistrons are located in bands 59.5, 69 and 72.5 in the Standard phylad of W. virgo. — The genomic organization of the 18S+26S rDNA cistrons, as shown by restriction enzyme analysis, is different in the two W. virgo phylads and there are also differences in organization between P196A and P196B. The pattern in P196B and that in the Boulder-Zanthus phylad suggest that they are related. As in the in situ analyses, the genomic organizations of the ribosomal cistrons in both W. virgo phylads are not simply the additive products of those in any known populations of P169 and P196. New repeat lengths indicative of segmental amplification events occur in particular clones of W. virgo. — Throughout the genus Warramaba the N-banding technique stains all bands containing 18S+26S and 5S rDNA cistrons. The Olert silver technique stains band 72.5 in the Standard phylad, but does not correlate with the locations of 18S+26S ribosomal genes.     

Cytogenetics of the parthenogenetic grasshopper Warramaba virgo and its bisexual relatives

Cloned highly repeated DNA sequences were used to investigate the origins of W. virgo. The chromosomal and genomic organization of these sequences in the parthenogenetic grasshopper W. virgo and its sexual relatives indicate that W. virgo has had two independent origins. Data from the cloned sequences together with that from rapidly renaturing DNA are consistent with a hybridization event between two sexual species for each origin of the parthenogenetic species. Previously published data on C-banding and other karyotypic features (White and Contreras 1981) strengthen the dual origin conclusion. — The cloned DNA sequences, pWv 1 and pWv 5, have differentiated northern and southern races in the sexual species P196. The southern race appears to have hybridized with another sexual species, P169, to give rise to the Boulder/Zanthus clones of W. virgo. The northern race of P196 may have crossed with a species similar to P169 to give rise to the remaining W. virgo clones which are now present in both eastern and western Australia. White (1980) proposed that the origin of W. virgo was in western Australia and that the eastern populations were established by migration. Consistent with this hypothesis is our finding that the cloned DNA sequences have identical genomic and chromosomal organisation in populations of W. virgo in the two disjunct areas.Respectfully dedicated to Professor Wolfgang Beermann on his 60th birthday, in recognition of his great contributions to modern cytogenetics     

Cytogenetics of the grasshopper Moraba scurra

Summary The cytology of a male individual of the grasshopper Moraba scurra which was heterozygous for a complex translocation involving breaks in four different, non-homologous, chromosomes, is described. Chains of up to eight chromosomes occur at first metaphase. Certain chromosomes which never normally show more than a single chiasma form two chiasmata with a fairly high frequency in this individual.Supported by Public Health Service Grant No. RG7212-Cl, from the Division of General Medical Sciences, U.S. National Institutes of Health.     

Finding of Bov-B LINE retroelement in parthenogenetic and bisexual lizard species of the genus Darevskia (Lacertidae)

The Bov-B LINE retrotransposon was first discovered in Ruminantia and was long considered to be specific for this order. Later, this mobile element was described in snakes and some lizard species. Analysis of phylogenetic relationships of Bov-B LINE elements from different ruminants, snakes, and lizard species led to the suggestion on horizontal transfer of this retrotransposon from Squamata to Ruminantia. In the Squamata group, Bov-B LINE element was found in all snakes and some lizard species examined. The element was not detected in the genomes of some species of the genera Lacerta and Podarcis. In the present study, using PCR amplification and sequencing of PCR products, Bov-B LINE element was identified in the genomes of parthenogenetic and bisexual species of the genus Darevskia (Lacertidae), as well as in such species as Lacerta agilis and Zootoca vivipara, where this retrotransposon had not been not detected before.     

Finding of Bov-B LINE retroelement in parthenogenetic and bisexual lizard species of the genus Darevskia (Lacertidae)

The Bov-B LINE retrotransposon was first discovered in Ruminantia and was long considered to be specific for this order. Later, this mobile element was described in snakes and some lizard species. Analysis of phylogenetic relationships of Bov-B LINE elements from different ruminants, snakes, and lizard species led to the suggestion on horizontal transfer of this retrotransposon from Squamata to Ruminantia. In the Squamata group, Bov-B LINE element was found in all snakes and some lizard species examined. The element was not detected in the genomes of some species of the genera Lacerta and Podarcis. In the present study, using PCR amplification and sequencing of PCR products, Bov-B LINE element was identified in the genomes of parthenogenetic and bisexual species of the genus Darevskia (Lacertidae), as well as in such species as Lacerta agilis and Zootoca vivipara, where this retrotransposon had not been not detected before.     

Cytogenetics of the hybrids of three members of the grasshopper genus Vandiemenella (Orthoptera: Eumastacidae: Morabinae)

Certain members of the grasshopper genus Vandiemenella (earlier referred to as the viatica group) form a group of chromosomally distinct taxa with contiguous distributions through south-eastern Australia. Meiosis of hybrids between three of the members that occur on Kangaroo Island, South Australia, was studied. The three members were the taxonomically undescribed form P24(XY) and two chromosomal races of V. viatica. Hybrids were either collected from narrow overlap zones (viatica ₁₉-viatica ₁₇, viatica ₁₉-P24 (XY), P24(XY)-vitica ₁₇) or were reared from grasshoppers collected from areas close to the overlap zones. — Chromosomal heterozygotes were found in all three overlap zones, but were frequent only in the overlap zone of P24(XY) and viatica ₁₇. Male chromosomal heterozygotes of P24(XY) and viatica ₁₉ and of viatica ₁₇ and viatica ₁₉ had abnormalities of meiosis, such as asynapsis (or desynapsis), which were generally associated with the chromosomes that distinguish the taxa. These abnormalities would be expected to lead to a reduction of fertility and so may contribute to reproductive isolation. Analysis of embryos from female heterozygotes of P24(XY) and viatica ₁₇ indicated that similar abnormalities of meiosis occurred in these females. Very few meiotic abnormalities, however, were seen in the corresponding male hybrids. The structurally different X chromosomes of female hybrids of P24(XY) and viatica ₁₇ were recovered in a distorted ratio in their embryos. The distortion was only slight and would not be expected to be of importance in the evolution of the group, particularly since the distortion favoured the original rather than the derived type of chromosome.     

Home ranges of parthenogenetic and bisexual species in a community of Darevskia lizards (Reptilia: Lacertidae)

We analysed the home ranges of a community of Darevskia rock lizards composed of a bisexual species (D. valentini), two parthenogens (D. armeniaca and D. unisexualis), and two backcross forms between bisexual and unisexual forms. We estimated home range areas of ink-marked, GPS-located lizards using Minimum Convex Polygon (MCP) and 95% of the locations for those individuals with five or more sightings. The bisexual D. valentini was the species with the largest home ranges, distances travelled, and the most intersections. No differences between unisexual species and backcrosses were recorded for any comparison. In males, home range size and perimeter were related to morphological characteristics. Contrary to what has been described in allopatry, unisexual species showed smaller home ranges and fewer overlaps than sympatric bisexual species. We tentatively suggest that the presence of potential bisexual partners might increase sexual competition among parthenogenetic females while differences in habitat use should also be considered.     

Genetic similarity and diversity of parthenogenetic and bisexual populations of the freshwater snail Melanoides tuberculata (Gastropoda: Prosobranchia)

Genetic variability and similarity were analysed in four parthenogenetic and five bisexual populations of the snail Melanoides tuberculata found in Israel. Electrophoretic studies of six enzymatic systems revealed 28 zones of activity. The average genetic identity between populations was low—0.725. A particularly low similarity (0.628) was obtained between parthenogenetic populations, compared to the average of 0.822 observed among the bisexual ones. The percentage of fixed electrophoretic bands in parthenogens was 53.9% compared to 12.5% observed in bisexual populations. The diversity of parthenogenetic populations was found to be lower than those of bisexual. The amount of electrophoretic diversity between populations of the parthenogenetic group was found to be 80%, whereas within the bisexual group the diversity between populations was only 42%.     

Contrasting patterns of molecular diversity and Wolbachia infection in bisexual and parthenogenetic Strophosoma weevils (Coleoptera: Curculionidae)

It has been postulated that parthenogenesis in weevil species is of hybrid origin, but some have speculated that Wolbachia infection plays a role through the modification of host breeding systems. Here we focus on Strophosoma weevils, which are known to be pests in young forest stands. Using molecular data, we investigated the diversity of the two most common Strophosoma species in Europe: S. capitatum, which reproduces bisexually, and S. melanogrammum, which is parthenogenetic. Also researched were their associations with the endosymbiotic bacterium Wolbachia. These species of weevil were found to be clearly distinguishable based on their mitochondrial DNA, with the bisexual taxa being more diverse. However, the nuclear DNA divergence of the two species was very low, and the parthenogenetic taxon was found to be heterozygous. Wolbachia infection was detected in all individuals of the S. melanogrammum populations and less than half of the S. capitatum populations. Moreover, multiple Wolbachia strains were found in both taxa (two in the former and three in the latter). The results of this research suggest that parthenogenesis in this genus is of hybrid origin and that Wolbachia could have played a role in speciation of these weevils.     

Pollen morphology of Trigonostemon and its relatives (Euphorbiaceae)

The pollen of Trigonostemon and the related genera Dimorphocalyx, Ostodes, Tritaxis and Jatropha (outgroup) has been studied with light microscopy, and scanning and transmission electron microscopy. The two major pollen types within Trigonostemon correlate well with macromorphological characters. Species belonging to the Trigonostemon reidioides type have pollen with ‘croton pattern’ ornamentation, a pistil with deeply divided stigmas (to at least half the length of the stigma arm) and stamens with a protruding appendage on the connective, while species of the Trigonostemon verrucosus type have verrucate (to almost gemmate) pollen, stigmas that are shortly cleft and stamens without an appendage on the connective. Dimorphocalyx, Ostodes, Tritaxis and Jatropha (outgroup) have similar pollen morphology, while Trigonostemon deviates from these genera in the absence of the ‘vertically’ striate ornamentation on the subunits. Therefore, when compared with an existing phylogeny of the Euphorbiaceae, the pollen characters of Trigonostemon appear to be derived. Moreover, because the ‘croton pattern’ ornamentation itself is widely shared by the ‘inaperturate crotonoids’, the loss of that structure in the Trigonostemon verrucosus type pollen is considered a further apomorphy.     

Viola hirta (Violaceae) and its relatives in Norway

Viola hirta is widespread in most of Europe but rare in Norway, occurring in four isolated enclaves around the Oslo Fjord. Allozymic and morphological patterns of variation were studied in 320 individuals of Viola hirta (2n = 20) and its relatives V. collina (2n = 20), V. odorata (2n = 20), and V. suavis (2n = 40) from 22 Norwegian populations. Ongoing introgression from both V. collina and V. odorata , as hypothesised by several authors, was not supported by our data. All hybrids proved to be primary ones and, apparently, introgression is rare. However, allozyme markers suggested that V. hirta may have been affected by ancient introgression from V. collina and possibly also from the East European V. ambigua (2n = 40) during its evolutionary history. Geographical differentiation, presumably relatively recent and post-glacial, was evident in V. hirta. Variation occurred also between and within V. odorata populations, most likely a consequence of independent introductions by man. No variation was observed in V. collina or in V suavis. A key to the Norwegian species of subsection Viola and their primary hybrids based on morphological characters is provided in Flora Nordica note No. 26.