The B chromosome polymorphism of the grasshopper Eyprepocnemis plorans in North Africa: III. mutation rate of B chromosomes

B chromosome variation in nine Moroccan populations of the grasshopper Eyprepocnemis plorans was analysed for 3 consecutive years. In addition to B1, which was the predominant B chromosome in all nine populations, we found 15 other B variants, albeit at very low frequency. Eight variants were found in adults caught in the wild, four appeared in adults reared in the laboratory and seven were found in embryo progeny of controlled crosses between a 0B male and a B-carrying female. Some variants were found in more than one kind of material. At least the seven B variants that appeared in embryo progeny of females carrying a different B type arose de novo through mutation of the maternal B chromosome. The mutation rate of B chromosomes was 0.73%, on average, which explains the high variety of morphs and banding patterns found. The most frequent de novo mutations observed in these chromosomes were centromere misdivision with or without chromatid nondisjunction, which generates iso-B-chromosomes or telocentric Bs, respectively, as well as translocations with A and B chromosomes and deletions. But the whole variation observed, including that found in adult individuals, suggests that other mutations such as duplications, inversions and centric fusions do usually affect B chromosomes. Finally, B chromosome mutation rate was remarkably similar in both Moroccan and Spanish populations, which suggests that it might be dependent on B chromosome intrinsic factors.     

The B-chromosome polymorphism of the grasshopper Eyprepocnemis plorans in North Africa: II. Parasitic and neutralized B1 chromosomes

The transmission of the B1 chromosome through females has been analysed in three Moroccan populations (Smir, SO.DE.A. and Mechra) of the grasshopper Eyprepocnemis plorans. We analysed transmission ratio (kB) variation at two levels: intra-individual (to test female age effects) and inter-individual (to test for A chromosome effects). In 81.8% of females, kB did not differ among successive egg-pods, suggesting no effect of female age. The remaining females (18.2%), showed significant differences in kB values among egg-pods, but without clear temporal patterns. In Smir, kB ranged between B elimination (0.244) and B accumulation (0.689) but there was no net accumulation (mean +/- s.e. = 0.463 +/- 0.045). In SO.DE.A., all females analysed transmitted B1 at a Mendelian rate, with a mean kBequal to 0.512 +/- 0.020. In Mechra, kB ranged from 0.341 to 0.972, with mean kB (0.575 +/- 0.029) showing a net B accumulation. All these observations suggest that the B1 chromosome could be at a drive-suppression stage in Smir and Mechra, but that it has already been neutralised in SO.DE.A.     

Spatio-temporal dynamics of a neutralized B chromosome in the grasshopper Eyprepocnemis plorans

Spatial and temporal patterns of frequency variation for a neutralized B chromosome in the grasshopper Eyprepocnemis plorans were analyzed along six transects in the east of Spain to explore possible factors affecting the population dynamics of this polymorphism. Three parameters were employed to quantify B frequency: prevalence, load and mean frequency. Of them, load seemed to be the less sensitive parameter, probably due to its small range of variation. Prevalence, however, shows ample variation, but the mean frequency of B chromosomes per individual is the best parameter to characterize B frequency. Only river transects revealed significant differences among populations, and the use of two geographic explicit approaches (Mantel test and distograms) revealed significant isolation by distance (IBD), especially at the Segura River mouth, presumably due to low gene flow and drift. No temporal trend was found in the Segura River transects, which is consistent with the slow changes in B frequency expected during the random walk for neutralized B chromosomes. But these transects showed a clear spatial pattern, with B1 showing lower frequency in the upper course of this river. The present results provide the first empirical evidence of IBD in the evolution of a neutralized B chromosome, and support the notion that B dynamics at this evolutionary stage is best explained by a metapopulation approach.     

Evolution of a complex B-chromosome polymorphism in the grasshopper Eyprepocnemis plorans

B-chromosome systems in several Spanish natural populations of Eyprepocnemis plorans are reported. The geographical distributions of the fourteen types of B chromosomes, which were classified according to size, C-banding pattern, and meiotic behaviour, are described. The results indicate a common origin for most types and a possible independent origin for a few secondary B variants. The origin and mechanisms of evolution of this B chromosome polymorphism are discussed.     

Evolutionary dynamics of a B chromosome invasion in island populations of the grasshopper Eyprepocnemis plorans

Four natural populations of the grasshopper Eyprepocnemis plorans in the Mallorca island were analysed for several years revealing the recent invasion of the B₁ chromosome from the south‐west part of the island (Palma region) towards the north and to the east. In only 10 years, the mean number of Bs in the northern population at Pollença increased from 0.053 to 0.692. Therefore, B chromosome invasion seems to be very rapid and has recently arrived to the north of the island. The south‐west (close to Palma) is the most likely point at which B invasion started in the Mallorca Island. Finally, the number of B chromosomes was significantly associated to an increase in chiasma frequency (and thus recombination) in A chromosomes.     

The B-chromosome system of the grasshopper Eyprepocnemis plorans subsp. plorans (Charpentier)

Supernumerary chromosomes of two types have been observed in the grasshopper Eyprepocnemis plorans subsp. plorans. One of these (the B-type) is similar in size to an S autosome; the other is smaller (B-type). Both are telocentric and mitotically stable. The frequencies of individuals with the B-type supernumeraries in five natural populations were 56, 56, 70, 71 and 30 per cent respectively. The equivalent levels of the B-type supernumerary were 0, 0, 13, 3 and 0 per cent respectively. Because of the relative infrequency of the B-type only the B-type has been studied in detail. In males with 1B, anaphase I segregation of X and B was random in four populations but non-random in that from Otivar. Here the B was distinctive in having a secondary constriction near the centromere. A study of chiasma frequency among A-chromosomes revealed that the B-type supernumerary increases significantly both the mean chiasma frequency and the between-cell variance. A comparison of body morphometrics failed to reveal any effect of these B-chromosomes on the exophenotype.     

A single, recent origin of the accessory B chromosome of the grasshopper Eyprepocnemis plorans

B chromosomes are dispensable chromosomes found in >2000 eukaryotic species, usually behaving as genomic parasites. Most B chromosomes seem to be made up of the same kind of DNA sequences present in the A chromosomes. This sequence similarity makes it difficult to obtain specific molecular probes that may permit B-presence diagnosis without cytogenetic analysis. We have developed a sequence-characterized amplified region (SCAR) marker for B chromosomes in the grasshopper Eyprepocnemis plorans, which specifically amplifies a 1510-bp DNA fragment exclusively in B-carrying individuals. Fluorescent in situ hybridization and fiber FISH analyses showed that this marker is a tandemly repeated DNA sequence closely intermingled with 45S rDNA. PCR reactions showed the presence of SCAR-like sequences in the A chromosomes, but in two separate fragments, supporting the intraspecific origin of B chromosomes in this species. SCAR marker DNA sequence showed to be identical in B chromosome variants from several localities from Spain and Morocco, and it was very similar to those found in B chromosome variants from Greece and Armenia. This strongly suggests that this sequence was already present in the ancestral B chromosome of this species. In addition, the scarce sequence variation observed among several B variants from very distant populations suggests either a functional constraint or, more likely, a recent and unique origin for B chromosomes in this species.     

The B chromosomes of the grasshopper Eyprepocnemis plorans and the intragenomic conflict

The grasshopper Eyprepocnemis plorans harbours an extremely widespread polymorphism for supernumerary (B) chromosomes, which is found in almost all circum-Mediterranean and Caucasian populations hitherto analysed. B chromosomes in this species have been shown to evolve through several stages of parasitic and near-neutral nature, presumably because of an arms race between the standard (A) and B chromosomes. This intragenomic conflict can either be solved with the extinction of the neutralised B chromosome or, more interestingly, with the replacement of the neutralised B by a mutant version being parasitic again and thus prolonging B chromosome life. This species thus provides a complete view of the long-term life-cycle of parasitic B chromosomes.     

Population dynamics of a selfish B chromosome neutralized by the standard genome in the grasshopper Eyprepocnemis plorans

Effects of the B chromosome polymorphism of the grasshopper Eyprepocnemis plorans were analyzed in two natural populations. Postmating sexual selection, female fertility, and survival were studied. The B chromosome lacks drive and has no detectable effects on fitness. A neutral B cannot invade a population and establish a polymorphism, but the confidence limits on our estimates cannot exclude the possibility that the polymorphism is maintained by a balance between weak drive and weak selection against individuals with two and three B's. However, other lines of evidence favor the following model of the dynamics of the B in E. plorans. In a newly invaded population, the B has substantial drive, but the evolution of drive suppressor genes in the A chromosomes neutralizes the B drive so that it becomes near-neutral and begins a random walk toward extinction by stochastic loss. Because the B is common by the time drive disappears, the random walk is likely to continue for a long time. If in the course of the random walk a variant B with greater drive appears, then it will displace the original variant, and a new cycle of drive suppression and drift to extinction occurs. A simulation model of this process suggested that the mean time to extinction is proportional to the two-thirds power of the population size; it is much less affected by subpopulation size or the number of populations in a subdivided population.     

Dynamics of sperm storage in the grasshopper Eyprepocnemis plorans

Abstract. Sperm storage duration in the grasshopper Eyprepocnemis plorans Charpentier (Orthoptera, Acrididae) was found to be 58.5 days but with a wide between-female variation (from 26 to 113 days). One copulation was enough to fertilize two to eight egg-pods and to produce 130 eggs (28–313) and 119 embryos (28–290), on average. Sperm availability, however, decreased progressively with time, so that the majority of females spent a long final period of their lives without laying any pod. Finally, sperm storage duration was positively correlated with clutch size and total production of eggs and embryos.     

Undertransmission of a supernumerary chromosome segment through heterozygous females possessing B chromosomes in the grasshopper Eyprepocnemis plorans.

The transmission ratio (ks) for a supernumerary chromosome segment was studied in a total of 54 heterozygous females collected from two Spanish natural populations of the grasshopper Eyprepocnemis plorans. Our analysis clearly demonstrated that ks is negatively dependent on the number of B chromosomes in the female. The possible mechanisms by which B chromosomes may cause undertransmission of the supernumerary segment, and the implications of this phenomenon for the maintenance of this extra chromosome segment, are discussed.     

The effect of B chromosomes on mating success of the grasshopper Eyprepocnemis plorans

The mating ability of E. plorans was tested in laboratory conditions in six experimental units composed of ten males and fifteen females during 31 days. When significant differences were found (three from the six cages, and in totals) they involved a decrease of matings involving males with B chromosomes. The same tendency seems to exist in females, but to a lesser extent, so that a significant effect is only detected when the totals are considered. Accessory chromosomes also delay, in both sexes, the occurrence of the first mating. No mating preferences depending on the number of Bs were detected.     

The b chromosome polymorphism in Armenian and Turkey populations of the grasshopper Eyprepocnemis plorans plorans (Charpentier) (Orthoptera, Acrididae, Eyprepocnemidinae)

Karyotype analysis of the grasshopper Eyprepocnemis plorans samples derived from Armenian and Turkey populations was performed using C-banding technique. Chromosome polymorphism was associated only with B chromosomes that were revealed in all studied populations. Six new B chromosome morphotypes were described. Four morphotypes were found only in Armenian populations. One morphotype was revealed only in Turkey populations. One B chromosome morphotype was present in Armenian and Turkey populations. B chromosomes derived from Asian populations consisted mostly from C-positive regions. In some of the B chromosomes small C-negative regions were also registered. Morphotypes of the B chromosomes derived from Armenian and Turkey populations drastically differed from the B chromosomes described in the Iberian Peninsula and North African populations. In contrast to the B chromosomes from Armenian and Turkey populations the B chromosomes from Spain populations contained C-positive and C-negative regions alternated in their arms.     

Parallel effects of a B chromosome and a mite that decrease female fitness in the grasshopper Eyprepocnemis plorans

The effects of a genomic parasite (a B chromosome) and an ectoparasite (a mite) on the fitness of the host (the grasshopper Eyprepocnemis plorans) have been analysed in 60 experimental females. These two parasites differ in their infectious transmission mode. B chromosomes are vertically transmitted from host-parents to offspring, but mites are horizontally transmitted from one grasshopper to another within the same generation. The transmission mode can influence the virulence of these parasites, so that it should be expected that B chromosomes would be less virulent than mites. However, as mite transmission is linked to host mobility, some attenuation is also expected. Four egg pods were analysed from each female, the first two egg pods were laid after a mating and the remaining two were not preceded by a mating. The results show that B chromosomes severely decrease the proportion of eggs containing an embryo (egg fertility), mainly from the second egg pod onwards. Mites also decrease egg fertility but, in addition, they produced a decrease in the rate of embryo production over time (embryo productivity), which might be derived from both the fertility decrease and a slight delay in egg production. The analysis of the relative effect of both parasites suggests that they have a synergistic effect on embryo clutch size and egg fertility. Possible mechanisms for the observed effects are discussed.     

Spontaneous translocations between B chromosomes and the normal complement in the grasshopper Eyprepocnemis plorans

A translocation between a B chromosome and a medium-size autosome and a centric fusion between the B and the X chromosome were found in two different natural populations of Eyprepocnemis plorans. The cytological behaviour of both interchanges is described and discussed. These interchanges are highly asymmetrical and may be the beginning of an integration of B chromosomes into the genome of this species.     

Detection of B chromosomes in interphase hemolymph nuclei from living specimens of the grasshopper Eyprepocnemis plorans

The two most important evolutionary properties of B chromosomes are their transmission rate (which suggests their selfishness when significantly higher than 0.5) and their net effects on carrier fitness (usually negative for parasitic Bs). The study of transmission rate unavoidably requires the analysis of many controlled crosses in order to accurately measure population average transmission rate. Therefore, getting a marker closely associated to B presence is of crucial importance to alleviate the load of performing many useless crosses between lacking B individuals. After investigating several cytogenetic techniques on several tissues that may be sampled without drastically damaging live specimens of the grasshopper Eyprepocnemis plorans, we report here the excellent results provided by the CMA3 fluorescence and C-banding techniques applied to hemocyte nuclei. These cells may be easily obtained from both males and females and provide information on B presence even during the interphase stage. The two cytogenetic techniques take advantage of the heterochromatic nature of the B chromosomes, so that Bs made predominantly of ribosomal DNA are revealed by CMA3 as bright bodies in the interphase hemocytes, and Bs mostly made of satellite DNA are visualized by C-banding as intensely stained bodies in these cells.     

Gene expression changes elicited by a parasitic B chromosome in the grasshopper Eyprepocnemis plorans are consistent with its phenotypic effects

Chromosoma - Parasitism evokes adaptive physiological changes in the host, many of which take place through gene expression changes. This response can be more or less local, depending on the organ...