Chromosome organisation in the Australian plague locust Chortoicetes terminifera

In Chortoicetes terminifera 45 independently-occurring B-chromosomes were tested and 23 distinct banding variants were detected with either G- or C-banding; six types were found more than once. In particular the Type I banding morph was detected 12 times indicating that individuals carrying this type may be under a different regime of selection compared with individuals bearing other types of banding morph; or the Type I may be subjected to a higher rate of meiotic drive in either or both sexes than other types. Also the Type I appeared to be obviously related to four other banding morphs whereas most types were not obviously related to any other banding morphs, but a few were similar in banding pattern to one or two other types. Three types of B-chromosomes were found in three or more different populations. A relatively high frequency of the Type I banding morph was found in one population, which was probably mainly composed of non-migratory individuals, and also in a laboratory-raised population. The most likely mechanisms for small changes in the banding sequence of the B-chromosomes are three-break insertions which are often indistinguishable from inversions. Rearrangements which add or delete bands, or sequences of bands, to or from B-chromosomes are probably the result of exchanges which are now known to take place in rare individuals with two B-chromosomes. The most distal region of all the banding morphs of the B-chromosome in C. terminifera, plus a short interstitial region in some types, is not late-replicating and has the banding characteristics of euchromatin. The rest of the chromatin of the B-chromosomes is heterochromatic and is the latest replicating heterochromatin in the whole genome. It consists of G-bands, which are also deeply stained with C-banding, and alternating G-interbands, which in turn are stained grey with C-banding. Both of these staining combinations are seen in heterochromatin of the normal complement. The heterochromatin of the B-chromosomes is condensed throughout 1st meiotic prophase in the male and in all somatic interphase nuclei where it can be quickly detected using the G-banding technique. The B-chromosome has a relatively constant, acrocentric morphology with a tendency to increase of length of the long arm as band numbers increase. Isochromosomes of the long arm have been seen only in laboratoryraised embryos. From egg pods with significantly fewer than expected B-chromosomes it is strongly suggested that more than one male may fertilize the eggs in a single pod.     

Chromosome organisation in the Australian plague locust,Chortoicetes terminifera

In Chortoicetes terminifera, G-banding, produced by the trypsin treatment of air-dried slides followed by Giemsa staining, leads to light staining gaps at the secondary constrictions on autosomal pair 6 and regions proximal to the centromere on the long arms of pair 4. The variable short arms of two of the three smallest pairs were usually flared and lightly stained after treatment. In contrast to the relatively minor response of the normal chromosome set to G-banding, the large supernumerary chromosomes of C. terminifera show a spectacular series of dark bands alternating with lightly stained gaps. Two G-band variants of the B-chromosome were found in a laboratory stock. These patterns of G-banding are discernable both at mitosis in adults and embryos of both sexes and at all stages of male meiosis. Some regions which are gaps after G-banding appear as dark bands after C-banding. Consequently the supernumerary chromosome is mainly darkly stained with C-banding. In addition the centromeres and some telomeres are C-banded along with narrow interstitial bands and polymorphic heterochromatic blocks. — C-banding was not always successful, the technique often yields a mixture of G- and C-banding. The disparity of banding between the normal complement and the B-chromosome implies that whatever the source of origin of the B it has undergone spectacular changes in organisation since its origin.     

Indigestibility of plant cell wall by the Australian plague locust, Chortoicetes terminifera

The plant cell wall may play an important role in defence against herbivores since it can be both a barrier to, and nutrient diluter of, the easily digested cell contents. The aim of this study was to investigate the digestibility of the cell wall of three grasses, Triticum aestivum L., Dactyloctenium radulans (R. Br.) Beauv., and Astrebla lappacea (Lindl.) Domin, by the Australian plague locust, Chortoicetes terminifera Walker (Orthoptera: Acrididae, Acridinae) as determined by the Van Soest method [ Van Soest PJ, Robertson JB & Lewis BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74: 3583–3597]. Determination of plant cell wall digestion by locusts required a precise methodological procedure to determine both the exact intake and the concentration of cell wall in the diet and the faeces. Plant cell wall determination is affected by the particle size distribution of the dried plant material. All three grasses differed in the percentage of cell wall per gram dry matter and the proportions of hemicellulose, cellulose, and acid‐detergent sulphuric lignin within the cell wall. The locust was unable to digest the cell wall of any of the grasses. Thus, plant cell walls are a mechanical barrier hindering locusts assimilating nutrients. That is, access, rather than nutrient concentration per se, may be limiting nutrient factor.     

Eight polymorphic microsatellite loci for the Australian plague locust, Chortoicetes terminifera

Few population genetics studies have been carried out on major locust species. In particular, an understanding of the population genetic structure of the Australian plague locust, Chortoicetes terminifera, is lacking. We isolated and characterized eight polymorphic microsatellite loci in C. terminifera, and described experimental conditions for polymerase chain reaction multiplexing and genotyping these loci. The number of alleles per locus ranged from 11 to 29 and the expected heterozygosity ranged from 0.797 to 0.977. One locus was found to be X-linked. Results of cross-taxon amplification tests are reported in four species of the Oedipodinae subfamily.     

Behavioural phase polyphenism in the Australian plague locust (Chortoicetes terminifera)

Swarming and the expression of phase polyphenism are defining characteristics of locust species. Increases in local population density mediate morphological, physiological and behavioural changes within individuals, which correlate with mass marching of juveniles in migratory bands and flying swarms of adults. The Australian plague locust (Chortoicetes terminifera) regularly forms migratory bands and swarms, but is claimed not to express phase polyphenism and has accordingly been used to argue against a central role for phase change in locust swarming. We demonstrate that juvenile C. terminifera express extreme density-dependent behavioural phase polyphenism. Isolated-reared juveniles are sedentary and repelled by conspecifics, whereas crowd-reared individuals are highly active and are attracted to conspecifics. In contrast to other major locust species, however, behavioural phase change does not accumulate across generations, but shifts completely within an individual''s lifetime in response to a change in population density.     

Regulation of water ingestion by the locust, Chortoicetes terminifera: the effect of injections into the haemolymph

Adults of the locust Chortoicetes terminifera which had been deprived of water for 24 h took larger 'meals' of water than insects which had been deprived for 2 h. Water or dilute solutions of NaCl injected into the kaemcoel reduced the amount of water ingested by 24-h, deprived locusts. Injections of more concentrated solutions of NaCl (0.5 m) or glycerol (1.0m) increased the water intake of 2-h deprived locusts. Water injections into 24-h deprived locusts did not reduce their responsiveness to sucrose and it is concluded that haemolymph osmotic pressure plays an important role in determining the specific responsiveness of C. terminifera to water. The time course of the effects of injection indicates that the effects of haemolymph osmotic pressure on water responsiveness are mediated neurally. Injections of water into 24-h deprived insects caused a marked reduction in the sizes of their meals of seedling wheat. It is suggested that the mechanisms responsible for the regulation of water intake might limit total intake when locusts have access only to fresh vegetation of high water content.     

Fat body changes in a locust,Chortoicetes terminifera (Walker) (Orthoptera: Acrididae), parasitized by a nemestrinid fly

The fat body of Chortoicetes terminifera (Walker) (Orthoptera: Acrididae) parasitized by Trichopsidea oestracea (Westwood) (Diptera: Nemestrinidae) was hypertrophied and showed extensive cytological and functional changes. Lipid content (as a percentage of dry weight) was significantly higher in parasitized locusts. Regression of lipid content against parasitoid weight was quadratic in female hosts and linear (negative) in male hosts. The fatty acid composition of C. terminifera was not affected by parasitization. Parasitized male locusts had lower body weight than nonparasitized males and contained less soluble protein. Results indicated that egg development was inhibited in parasitized female locusts. These conditions may be related to changes in the host's fat body.     

Levels of ecdysteroids in diapause and non-diapause eggs of the Australian plague locust,Chortoicetes terminifera (Walker)

The levels of ecdysteroids during embryogenesis in Chortoicetes terminifera were measured by radioimmunoassay. Soon after laying, eggs laid by females reared under non-diapause conditions contained three times as much as those laid by females reared under diapause inducing conditions. In eggs incubated at 32°C which did not enter diapause there was a sharp rise in ecdysteroid levels which coincided with the formation of the first-instar cuticle, but this did not occur in eggs incubated at 20°C even though they developed and hatched normally. Comparisons are made with previous results in other locust species and the possible role of ecdysteroids in embryonic diapause is discussed.     

Analysis of the development of major plagues of the Australian plague locust Chortoicetes terminifera (Walker) using a simulation model

The development and decline of all the past major plagues of Chortoicetes terminifera in Australia has been analysed using all available locust data and a simulation model to estimate the course of events for periods when few locusts were reported. The model is based on the developmental biology of the locust and includes methods of estimating the duration of egg, nymphal and adult development and uses a pasture growth index and potential night displacement index to assess likely survival and migration. The analysis shows that five of the six plagues originated in the arid interior, where good spring and summer rain allowed two or three consecutive generations of successful breeding, before emigration to the agricultural country occurred in the late summer or early autumn. This is contrary to the idea held previously that most plagues developed from local breeding in established outbreak areas in the agricultural belt. The applications of these results, to improve population monitoring and control strategy by suggesting the best timing for surveys and when control would have most effect, are discussed.     

Aerial treatment of the Australian plague locust, Chortoicetes terminifera (Orthoptera: Acrididae) with Metarhizium anisopliae (Deuteromycotina: Hyphomycetes)

Between October 1999 and April 2000, nearly 4000 ha of nymphal bands and adult swarms of Chortoicetes terminifera (Walker) were aerially treated using a ULV oil formulation of strain FI-985 of Metarhizium anisopliae var. acridum. During the mild weather (maxima 22-30 degrees C) of spring (October), there was little change in nymphal bands during the first week but at all doses between 25-100 g (1-4 x 10(12) conidia) ha(-1), the bands rapidly declined 9-12 days after treatment reaching > 90% mortality by 14 days. Metarhizium persisted for some time as there was 50% mortality of locusts fed vegetation collected from the treated blocks seven days after treatment. Persistence was confirmed by the high mortality of bands that invaded from untreated areas and of nymphs that hatched on the plot five to seven days after treatment, though mortality was then delayed until early in the third week. During summer (January), temperatures were high (maxima 36-42 degrees C), and at all doses between 25 and 125 g (1-5 x 10(12) conidia) ha(-1), there was a rapid decline seven to ten days after treatment. By 12-14 days, there was a > 90% decline in numbers in most blocks which was confirmed by helicopter surveys two weeks after treatment that found very few adults within or near treated areas. Mortality was delayed in the high dose where there were blockages of spray equipment during treatment. The clear demonstration that Metarhizium can suppress small local populations of C. terminifera led to the limited operational use of Metarhizium on an organic farm and in a National Park where nearly 2500 ha of bands and swarms were treated. Continued research is needed to develop a commercially viable product so that Metarhizium can form a significant part of a programme of integrated pest management of locusts in Australia.     

Isolation from a marching band increases haemocyte density in wild locusts (Chortoicetes terminifera)

1. The density‐dependent prophylaxis hypothesis predicts that individuals in high‐density populations will invest more resources in immune defence than individuals at lower densities. 2. However, recent work suggests that this prediction may not apply to all situations; solitarious species may paradoxically have higher scores than crowded counterparts in certain immune assays. 3. To investigate the relationship between a key immune parameter and field population densities, the total haemocyte counts (THCs) of Australian plague locusts (Chortoicetes terminifera) from three population densities in Western Australia were compared. 4. THCs were negatively correlated with field population densities, and locusts removed from a marching band and kept in isolation had increased THCs relative to group‐housed controls. 5. These results demonstrate that immune investment can inversely relate to population density in field conditions. 6. We suggest that isolated locusts increase their haemocyte densities relative to crowded conspecifics in response to potentially greater exposure to parasitoids and nematodes.     

The role of pressure in controlling the entry of water into the developing eggs of the Australian plague locust Chortoicetes terminifera (Walker)

The normal internal hydrostatic pressure and the additional pressure necessary to rupture the egg shell was measured in the eggs of Chortoicetes terminifera, Newly laid white eggs burst at c 0.15 kg cm^-2, but after external tanning the chorion withstands c 0.5 kg cm^-2 when removed from its tanned foam ‘corset’ and 1.0 kg cm^-2 if left embedded in the egg pod material. Older eggs with formed cuticles often withstand 2.0 kg cm^-2 but yield at rather lower pressures if they develop ‘pin-holes’. As the OP of the egg contents always exceeds 7.7 kg cm^-2 the rigidity of the wall is clearly insufficient to permit the generation of high hydrostatic pressures capable of preventing water entry during the non-absorbing phases of development. Real hydrostatic pressures are lower than 0.06 kg cm^-2 in the young intact egg and reach only c 0.5 and 0.3 kg cm^-2, respectively, during the absorptive and post-absorptive phases of development. Several events contribute to the sigmoid form of the water uptake curve. Water is at first excluded by a permeability barrier associated with the chorion. Absorption is delayed until the yolk is completely enclosed by the serosal cell layer. After undergoing cleavage, the yolk is then rapidly mobilized to furnish precursors for cuticle synthesis; in consequence, the internal OP rises from δ 0.76d?K to 0.93d?K despite the massive inflow of water which is governed by the osmotic gradient. At blastokinesis the serosa becomes detached from the cuticle; cuticle deposition and yolk mobilization are halted, the OP falling rapidly to cδT 0.53d?K. The bulk entry of water then ceases. Any excessive hydrostatic pressures which develop later are relieved by the formation of self-sealing ‘pin-holes’.     

The effects of temperature and moisture on the inception of diapause in eggs of the Australian plague locust,Chortoicetes terminifera Walker (Orthoptera: Acrididae)

Chortoicetes terminifera lays a mixture of diapause and non-diapause eggs during the autumn months of March and April. Diapause eggs cease development in late anatrepsis when kept in moist soil at 32.5°C. These conditions favour rapid embryogenesis and hatching in non-diapause eggs. Significant reductions in the incidence of diapause were obtained when newly-laid eggs were kept for 40 days at either 15.5°C or 32.5°C in conditions that prevented the uptake of water. In eggs held for a similar period in moist soil at 15.5°C, diapause was virtually eliminated. These eggs did not develop to the stage at which water uptake occurs. It is suggested that the inception of diapause is dependent upon the occurrence of warm, moist conditions at the time of laying and that these requirements are an integral part of the overall inductive process, which also involves the temperature and photoperiod at which the parent insects are reared. A flow-diagram illustrating the development of diapause eggs in relation to temperature and moisture is discussed in relation to the survival of the species.     

The effects of temperature and photoperiod on the morphology of the egg-pod of the Australian plague locust (Chortoicetes terminifera Walker,Orthoptera: Acrididae)

Chortoicetes terminifera can detect absolute differences of both temperature and photo-period, as well as the directional change of these parameters. Insects subjected to long-day conditions, or to an increase in photo-period during development lay long straight pods, whereas those subjected to short days or to a decrease in photoperiod during development lay short pods, which are usually curved and/or laid obliquely. Temperature does not affect oviposition behaviour at long photoperiod, but at medium and short photoperiod, high temperature or an increase in temperature promotes the production of long pods, whereas low temperature or a decrease in temperature stimulates the production of short pods. When temperature and photoperiod change in opposite directions, the effects of photoperiod override those of temperature. The significance of these findings is discussed in relation to egg development and survival.     

Expression from two Drosophila promoters in embryos of the migratory locust

We have accomplished gene transfer into embryos of Locusta migratoria, the African migratory locust. Freshly oviposited eggs were injected with circular or linear plasmids containing the Drosophila hsp70 promoter and the choramphenicol acetyltransferase (CAT) reporter gene (hsp-cat), or with circular plasmid containing the Drosophila copia promoter fused to CAT (copia-cat). Southern blot analysis showed that the hsp-cat plasmid persisted extrachromosomally for at least 8 days after injection. There was no evidence for plasmid replication. Transient expression from the introduced promoters was determined by monitoring CAT enzyme activity. After injection of hsp-cat, activity was detected at varying levels in 6–8% of day 3 and day 9 embryos. Embryos injected with copia-cat, assayed on day 3, had a greater frequency but no higher level of expression. The described gene transfer system is promising for analysis of other promoters, including those of Locusta.