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.     

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.     

Parallel polymorphism for supernumerary segments in Chorthippus parallelus (Zetterstedt)

French populations of Chorthippus parallelus like those from Britain are polymorphic with respect to supernumerary segments on the M₇ and S₈ chromosomes. The pattern of polymorphism parallels that of British material on both morphological and behavioural grounds. It agrees also in its effect on mean cell chiasma frequency and the absence of any effect on between cell variance. Finally the agreement extends also to the distribution of karyotypes within populations, those of the S₈ showing a Hardy-Weinberg distribution whereas those of the M₇ show a deficiency of structurally heterozygous types. All these parallels imply a common origin for the British and French material, an origin which, so we must suppose, antedated the physical separation of Britain and France some 8,000 years ago.     

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.     

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.     

Segregation-distortion in the B-chromosome system ofTettigidea lateralis (say) (Orthoptera: Tetrigidae)

A marginal population ofTettigidea lateralis was found to be polymorphic with respect to a large, mitotically stable supernumerary (B) chromosome. Male and female individuals may carry one or two B-chromosomes. In the male sex the frequency of individuals with one B was 33.8% whereas that of 2B-carriers was 2.9 %. A comparison with a small sample of female individuals suggests similar frequencies of B-chromosome carriers in the two sexes. The pycnocity cycle of the B's is virtually identical to that of the X chromosome which is always distinguishable by virtue of its larger size and other structural details. Persistent heterochromatic associations between the B and the X, which may last until metaphaseanaphase I, lead to a preferential migration of the B with the X to the same pole in male carriers of a single supernumerary. This distortional segregation of the B-chromosome may produce a differential transmission of the supernumerary to the two sexes if the various types of gametes are equally functional. Achiasmate, persistent B-B associations in 2B individuals can also cause segregation-distortion. The two supernumeraries segregate to the same pole in approximately 1/3 of the spermatocytes, but their poleward movement relative to that of the X is random. Both –B and +B individuals show only a single chiasma per individual bivalent. However, the presence of a single B raises very significantly the frequency at which the chiasma forms at the extreme distal ends of the L₁-L₂ and M₃-M₄ autosomes. The effect on recombination exerted by the supernumeraries and the possible implications of the segregation-distortion system ofT. lateralis are discussed in the light of recent studies on comparable B-chromosome polymorphisms.Research supported by N.R.C. of Canada.     

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     

Cytogenetic studies on Chorthippus jucundus (Fisch.) (Orthoptera) III. The meiotic consequences of a spontaneous centric fusion

A spontaneous structural heterozygote originated from a centric fusion involving two acrocentric autosomes (M₅ and S₈) in a male Chorthippus jucundus (2n=16+X) was tested for the effects on chiasmata characteristics and the role that these could play in metaphase-I orientation of the trivalent. Results show non-existence of inter- or intra-chromosomal effects on chiasma frequency when compared with standard individuals, and displacement of the single chiasma on the S₈ to a distal position while no significant change is appreciated in the M₅ chiasma distribution. The presence of two chiasmata on the M₅ and one on the S₈ always induces a linear orientation of the trivalent. However, when two chiasmata are present (one on each chromosome) and independently of their location on each arm (proximal, interstitial of distal) any of the possible orientations (linear or convergent) can be attained. The frequency of linear orientation is higher than that expected at random in the resulting trivalent during metaphase-I and the rate of reorientation is very low.The meitotic behaviour of such a trivalent is discussed taking into account the already mentioned aspects together with the size of the fused element.     

Testing the efficiency of three 15N-labeled nitrogen compounds for indirect labeling of grasshoppers via plants in the field

In field studies of plant–insect herbivore interactions it is often difficult to establish which herbivore has fed on a particular plant. We investigated the suitability of three different ¹⁵N‐labeled nitrogen compounds (ammonium, nitrate, and glycine) for indirect marking of three grasshopper species [Omocestus viridulus (L.), Chorthippus parallelus (Zett.), and Chorthippus biguttulus (L.) (Orthoptera: Acrididae)] through labeling their food plants in the field. In two short‐term experiments grassland plots of 1 m² were separately labeled with either one of the different nitrogen compounds. Grasshoppers were caged on three food‐plant species [Dactylis glomerata L., Holcus lanatus L. (Poaceae), and Trifolium repens L. (Fabaceae)] present in these plots for 72 h. Significantly enriched δ¹⁵N values in grasshoppers were found in all plant/grasshopper combinations. Enrichment in grasshoppers was positively correlated with the enrichment of plants and labeling with nitrate resulted in highest ¹⁵N enrichment. In a long‐term experiment, individuals of C. biguttulus were placed in a cage covering an area of 1 m² for 37 days, with sampling of grasshoppers at regular intervals. δ¹⁵N values of the grasshopper and a common food plant, D. glomerata, increased steadily over time, up to 40‐fold by the end of the experiment. Our results demonstrate that ¹⁵N‐labeling of plants is an appropriate tool for the investigation of insect–plant interactions under natural conditions.     

Central nervous control of hindleg coordination in stridulating grasshoppers

 Stridulation in many gomphocerine grasshoppers is characterized by specific phase shifts between the two hindlegs as well as different movement patterns produced by the left and the right leg. The underlying neuronal excitation patterns are generated by networks on either side of the metathoracic ganglion. The role of the intraganglionic commissures in right-left coordination and the production of differing movement patterns was investigated by transecting the metathoracic ganglion mediosagittally in Omocestus viridulus, Chorthippus biguttulus and Chorthippus mollis. In all three species, after this operation both hindlegs produced the same pattern and no longer different movements. The effects of transection on coordination differed: rapid movement rhythms, like those typical of Ch. biguttulus and the vibratory parts of the song of Ch. mollis, on the two sides drifted with respect to one another. In contrast, the slow rhythms characteristic of O. viridulus and the song subunits of Ch. mollis were completely synchronized. It is inferred that in intact animals the pathways for coordination of the rapid stridulatory rhythms are exclusively intraganglionic, whereas the phase relations of the slow rhythms are additionally influenced by way of anterior right-left connections, perhaps within the suboesophageal ganglion. Accepted: 15 October 1996     

The cytogenetic systems of grasshoppers and locusts

Two new cases of supernumerary heterochromatic segments are described. One of these, found in a heterozygous state in eight males of Chorthippus jucundus, is present terminally on the short arm of the longest autosome. The other, present in a single male heterozygote of Trimerotropis tolteca modesta, is again terminally sited, this time on the long arm of the second smallest auto-some. Both segments are considered to be products of tandem duplication rather than translocation. The segment system of C. jucundus, like that of a number of other categories of heterochromatic material, produces a striking increase in mean cell chiasma frequency compared to basic homozygotes from the same population. It is argued that this effect of the segment on recombination provides a potential basis for selection leading to altered fitness. This, in turn, may well have determined the evolution of at least some forms of supernumerary material in natural populations.     

Postglacial expansion and genome subdivision in the European grasshopper Chorthippus parallelus

A noncoding nuclear DNA marker sequence (Cpnl-1) was used to investigate subdivision in the grasshopper Chorthippus parallelus and deduce postglacial expansion patterns across its species range in Europe. Investigation of the spatial distribution of 71 Cpnl-1 haplotypes and estimation of levels of genetic differentiation (K_ST values) between populations and geographic regions provided evidence for subdivision of C. parallelus into at least five major geographic regions and indicated that the French form of C. parallelus originated after range expansion from a Balkan refugium, Further evidence for subdivision of C. parallelus between Italy and northern Europe suggests that the Alps may have formed a significant barrier to gene flow in this grasshopper.     

Improvement of antagonistic capability of Trichoderma harzianum by UV-Irradiation for management of Macrophomina phaseolina

Antagonistic capability of Trichoderma harzianum was improved through UV-irradiation. Four different type of mutants, T. harzianum - M_a (Th-M_a), T. harzianum - M_b(Th-M_b), T. harzianum - M_c (Th-M_c), T. harzianum - M_d (Th-M_d) of T. harzianum and the parent strain (Th-P) were selected for further studies. Th-M_a and Th-M_b showed more antagonistic capability against Macrophomina phaseolina than its parent strain Th-P in dual culture. Biochemical analysis of these four mutants and the parent strain showed that Th-M_a releases higher level of two lytic enzymes i.e. chitinases and cellulases and Th-M_b produces more β-1,3-glucanase activity than the parent strain. Culture filtrate of Th-M_a also showed antifungal properties. Study of the competitive saprophytic ability (CSA) of these four mutants and the parent strain were also made. Th-M_a exhibited higher CSA than the parental isolate while Th-M_d had less CSA than all other mutants and the parent strain of T. harzianum.     

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.     

Identified descending brain neurons control different stridulatory motor patterns in an acridid grasshopper

During courtship sequences male grasshoppers of the species Omocestus viridulus successively perform with their hindlegs three different stridulatory movement patterns: ordinary stridulation, hindleg shaking and precopulatory movements. Microinjection of acetylcholine into protocerebral neuropil regions can either elicit complete courtship sequences or evoke one of the three motor patterns. Intracellular recordings and stainings revealed three types of descending brain neurons: B-DC-3, B-DC-4 and B-DC-5. All three types of interneurons have a medial axon position in the connectives. They cross the midline of the protocerebrum and exhibit a profuse arborization pattern within the medial dorsal protocerebral neuropil. Stimulation of each type of interneuron specifically elicits one particular motor pattern of courtship behaviour. Courtship of the grasshopper O. viridulus may therefore be controlled by successive activation of these descending brain neurons. Accepted: 27 September 1996     

Effects of different supernumerary segments on chiasma distribution of the polymorphic species Chorthippus jucundus (Orthoptera: Acrididae)

The analysis of 18 wild populations of the polymorphic species Chorthippus jucundus has revealed the presence of five distal extra segments. They differ in location, size and staining properties. Two of them (L₁H and L₁h) are located in the largest element and do not C-band, while the remainder (S₈H, S₈h and S₈h₁) appear in the shortest chromosome and show positive C-banding. Those located in the same chromosome differ basically in their size. However, any of these blocks induce a similar change in the pattern of chiasma distribution within the affected bivalent. Additionally four patterns of C-banding as regards the presence of distal bands in some autosomes have been detected in the different populations.     

The supernumerary segment system of Stethophyma

Three species of the genus Stethophyma carry supernumerary heterochromatic segments. The European species, S. grossa, has segments located close to the centromere on the S₁₁ chromosome pair, while the North American species, S. lineatum and S. gracile, have both interstitial and terminal segments on the S₁₀ and S₁₁ chromosomes. The latter species show a high degree of segment variation between individuals and the interstitial segments also show variation in size. The presence of segments on the S₁₀ and S₁₁ chromosomes, whether homozygous or heterozygous, modifies the pattern of chiasma distribution in these chromosomes when compared with that found in the basic homozygotes. When interstitial, they also lead to a high frequency of ring bivalents.Two points suggest that interstitially located supernumerary segments may prove to be more common than has previously been accepted. Firstly, combined equational and reductional segregation in unequal bivalents is only otherwise explicable in terms of chiasma formation in a short arm. Secondly the rod chromosomes of many Acridids may well be telocentric as in the case under study.It is proposed that these segments have arisen through a process of duplication with no evidence of interchromosomal movement.On educational leave from the Forest Research Laboratory, Canadian Forestry Service, Fredericton, New Brunswick, Canada.     

Neuropharmacological evidence for inhibitory cephalic control mechanisms of stridulatory behaviour in grasshoppers

In gomphocerine grasshoppers the neuromuscular patterns of stridulatory hindleg movements are produced by metathoracic rhythm generators under the control of cephalic command neurons. Injections of cholinergic agonists into the protocerebrum activate this command system which induces the performance of stridulatory sequences, resembling natural species specific movements. Injections of GABA, glycine and picrotoxin into the central protocerebrum of the species Omocestus viridulus, Chorthippus mollis and Ch. biguttulus revealed a contribution of inhibitory mechanisms to the control of the stridulatory behaviour. The experiments suggest that inhibition interferes with the cephalic command systems at three levels: (1) sustained inhibition through picrotoxin sensitive receptors acting on all command units while grasshoppers are at rest, and during stridulation on all command units except the one activating the pattern generators of the currently performed movements; (2) premature termination of song sequences, experimentally induced by injections of GABA and glycine; and (3) coupling of a timing mechanism that terminates a song sequence or its subunits with a particular movement pattern after specific durations. These results together with those from previous studies on the pharmacological activation of stridulatory behaviour suggest that a balance of inhibitory and excitatory inputs to the command system selects the appropriate song type and controls its performance. Accepted: 11 June 1998