The male genitalia of the fritillary butterfly Issoria lathonia (L.) were examined and reconstructed based on sagittal and horizontal sections. Nine intrinsic muscles were identified consistent with previous results. The retractor of the anal tube probably operates the "rectal plate", a large, sclerotised, arched plate present ventral to the rectum and dorsal to the phallus in all Issoria s. str. species. The function of the rectal plate is still largely unknown, but it has presumably an important function during copulation. The retractor of the phallus inserts on the phallus, and also on a small, ventral sclerite in the anellus. The retractor of the vesica is smaller in I. lathonia than its counterpart in other Argynnini and originates more centrally inside the phallus. The tergal sclerite, common in most Argynnini, has no attaching muscle and its evolutionary origin remains unclear. The presence of an intrinsic muscle (i3) originating on the tegumen and inserting on the valve in Argynnini cannot be confirmed here. Though generally absent in butterflies, this muscle has been reported once in the North American Argynnis subgenus Speyeria.

Urbanisation results in a marked modification of habitats and influences several ecological processes, some of which give rise to beneficial ecological services. Natural pest control, the effect of predators on prey is one of such services. We quantified changes in the incidence of predation with increasing levels of urbanisation using artificial caterpillars made of green plasticine. Potential predators can be identified by the "attack marks" they leave on these artificial caterpillars. We conducted this study from May to October 2010 around the city of Sorø (Zealand, Denmark), in forests along an urbanisation gradient (rural-suburban-urban). Artificial caterpillars were placed on the ground in order to obtain an estimate of the incidence of predation at ground level. Half (50%) of the 1398 caterpillars were "attacked" and 28.8% of the bites were those of chewing insects. We attributed the majority of these to carabids, the most common group of ground-active arthropods. Chewing insects exerted the greatest predation pressure in the original forest (52.1%), with lower values recorded in the suburban (10.1%) and urban (16.4%) forest fragments. Ants were responsible for only 4.7% of the attacks in forest, 11.3% in suburban and 16.4% in urban forest fragments. Mammals exerted the highest predation pressure in suburban habitats (22.2% vs. 4.9% in forest, and 8.1% in urban forest fragments).

A cytogenetic investigation was performed in eight species of the spittlebug genus Philaenus using silver-NOR (AgNOR)-banding and fluorescence in situ hybridization (FISH) with 18S rDNA and (TTAGG)_n telomeric probes. This is the first application of FISH technique in the Auchenorrhyncha, a suborder of the Hemiptera. FISH along with the rDNA probe revealed differences between species in the number and chromosomal location of major ribosomal RNA gene sites, the so-called nucleolar organizer regions (NORs). However, we found a lack of perfect correlation between the results of AgNOR-staining and rDNA-FISH in the detection of NORs. FISH with the telomeric probe confirmed that the chromosome ends of the Philaenus species are composed of the (TTAGG)_n nucleotide sequence, which is a common motif of insect telomeres.

Mated females of the small copper butterfly Lycaena phlaeas avoid harassment by males by closing their wings and concealing themselves when in the proximity of a con-specific butterfly. This wing-closing behaviour is less frequently exhibited by virgin females that are two days old or older (i.e., potentially receptive) than by mated females. During the first 2 days after emergence, females of L. phlaeas are sexually immature and unreceptive. To determine whether recently emerged virgin females try to avoid male harassment, age-related changes in the frequency of harassment-avoidance behaviour of virgin females were investigated. On the day of emergence, a high percentage of virgin females exhibited wing-closing behaviour. Over the following 2 days, however, the frequency of this behaviour declined sharply and then reached a constant low level. This observation supports the idea that the harassment-avoidance behaviour exhibited by virgin females of L. phlaeas depends on their receptivity.

Thelytokous parthenogenesis is a type of sex determination in which females are produced from unfertilized eggs. Genetic and endosymbiont-induced forms of thelytoky have been described in the Hymenoptera. Our study has revealed that Wolbachia, Cardinium, Spiroplasma and other endosymbionts are probably absent in Iberian populations of harvester ants, Messor barbarus and Messor capitatus (Hymenoptera: Formicidae: Myrmicinae) and are thus not involved as factors in the sex determination of these two species. Our results lend weight to previous suggestions that bacterial parthenogenesis induction in Hymenoptera is probably limited to the reproductive systems of chalcidoid and cynipoid parasitoid wasps (Hymenoptera: Apocrita: Chalcidoidea and Cynipoidea, respectively).

As urbanisation is set to continue, understanding the impact on wildlife becomes increasingly important if we are to be able to conserve biodiversity. As an excellent group of bioindicators, invertebrates can allow us to understand some of the forces in urban areas which impact upon biodiversity and wildlife populations. This paper discusses some of the trends in the abundance, diversity and richness of invertebrates related to urbanisation and the specific urban environmental and traffic factors which may be at play.

Sex ratio and chromosomes studies were performed on six of the seven species of genus Cyclocephala present in the Lesser Antilles. Most sex ratios based on light trapping are skewed, with a frequent, but not exclusive, strong excess of females. These unbalances vary from species to species and island to island for the same species. Comparing old (seventies) and recent captures, the imbalances seem to increase with time. All of the 72 karyotyped males were diploid: 20,XY. Of the 15 females studied, one (C. dominicensis) was triploid: 30,XXX, a condition that only occurs in parthenogenetic species of beetles. We conclude that parthenogenesis is progressively developing within different sexual populations of Cyclocephala in the Lesser Antilles, which accounts for the excess of females recorded there. We propose that preexisting recessive mutations are the cause and that island colonization, by preventing panmictic reproduction, favours the expression of these recessive mutations. This would account for the accelerated occurrence of new characters (speciation /sub-speciation process), including unusual modes of reproduction such as parthenogenesis, and that several genetically related populations express these characters independently on different islands.

Pensoft Series Faunistica No. 76. Pensoft Publishers, Sofia, Moscow, 2008 (Vol. 1-4), 2009 (Vol. 5). 3264 pp. ISBN 978-954-642-317-7 (HB) (Vols 1-5). Price EUR 600 (EUR 120 each Vol.).

O'Toole C. 2013: Bees A Natural History. Firefly Books, New York and Ontario, 240 pp., 125 colour photographs, 3 appendixes. ISBN 978-177085-208-2. Price GBP 30.00.

Whereas the classification of plant communities has a long tradition that of animal assemblages remains poorly developed. Here we propose a classification scheme for orthopteran communities based on regional "character species", "differential species" and "attendant species" at different levels of habitat complexity, which is also applicable to other insect groups. In this context there are three main points of special importance: (i) the geographical reference area, (ii) the hierarchical spatial level (e.g. habitat complex, habitat and microhabitat) and (iii) precise constancy criteria for the definition of character species and differential species. We develop this new approach using a study on orthopteran communites of central German semi-dry calcareous grasslands. Within this habitat, we describe seven structural types that are characterized by specific orthopteran communities. For the arrangement of the structural types several environmental parameters (e.g. height and density of vegetation) were collected. Orthopteran densities were sampled at 80 sites using a biocoenometer (box quadrat). Regional character species of semi-dry grasslands include Myrmeleotettix maculatus, Metrioptera brachyptera, Stenobothrus lineatus and Tetrix tenuicornis. Within this habitat, Chorthippus parallelus, Metrioptera roeselii, Omocestus viridulus, Pholidoptera griseoaptera and Tettigonia viridissima were designated as differential species for particular structural types. Furthermore, Tettigonia cantans and Tettigonia viridissima act as altitudinal differential species. Chorthippus biguttulus is the only attendant species with high constancy values in all structural types. This classification is a powerful tool for arthropod conservation, since it allows one to determine community completeness of very important and threatened habitats, like semi-dry calcareous grasslands.

To date, thousands of microRNAs (miRNAs) and their precursors (pre-miRNAs) have been identified in insects and their nucleotide sequences deposited in the miRBase database. In the present work, we have systematically analyzed, utilizing bioinformatics tools, the featural differences between human and insect pre-miRNAs, as well as differences across 24 insect species. Results showed that the nucleotide composition, sequence length, nucleotides preference and secondary structure features between human and insects were different. Subsequently, with the aid of three available SVM-based prediction programs, pre-miRNA sequences were evaluated and given corresponding scores. Thus it was found that of 2633 sequences from the 24 chosen insect species, 2229 (84.7%) were successfully recognized by the Mirident classifier, higher than Triplet-SVM (72.5%) and PMirP (72.6%). In contrast, four species, including the domesticated silkworm, Bombyx mori L., the fruit fly, Drosophila melanogaster Meigen, the honeybee, Apis mellifera L. and the red flour beetle, Tribolium castaneum (Herbst), were found to be largely responsible for the poor performance of some sequence matching. Compared with other species, B. mori especially showed the worst performance with the lowest average MFE index (0.73). Collectively these results pave the way for understanding specificity and diversity of miRNA precursors in insects, and lay the foundation for the further development of more suitable algorisms for insects.

LANCASTER J. & DOWNES B.J. 2013: AQUATIC ENTOMOLOGY. First Edition. Oxford University Press, Oxford, 296 pp. ISBN 978-0-19-957321-9 (hbk.), ISBN 978-0-19-957322-6 (pbk). Prices: EBook: USD 48.29, Hardcover: USD 95.98, Paperback: USD 62.83.

Despite its limitations, ethanol remains the most commonly used liquid for the preservation of arthropod specimens and their DNA in the field. Arthropod ecology and taxonomy have witnessed a substantial increase in the use of various trapping and molecular methods in the past two decades. However, the methods of collecting and the preservation liquids most widely used in arthropod traps do not properly preserve DNA. Trap-collected specimens are typically of limited utility for molecular studies due to the poor preservation of DNA. A stable and cheap substance that can be used to trap arthropods in and preserve their DNA is therefore needed. Here we test whether (i) 2% SDS, 100mM EDTA, (ii) 1% SDS, 50mM EDTA and (iii) 0.66% SDS, 33mM EDTA can preserve DNA of small and medium-sized beetles for one, four and eight weeks. Preservation of DNA was tested using PCR amplification of parts of the mitochondrial cytochrome c oxidase I (Cox1) and nuclear 28S rRNA genes. All the solutions tested preserved DNA for at least up to eight weeks and we recommend 2% SDS and 100mM EDTA as a cheap, stable and easily transportable alternative to ethanol for preserving specimens and their DNA collected in the field. This solution is also suitable for using as the collection and preservation liquid in arthropod traps.

Little is known about the life cycle of Rhembobius parasitoids (Hymenoptera: Ichneumonidae). Known host records are mainly from saprophagous species of hoverflies of the tribe Eristalini (Diptera: Syrphidae), whose larvae develop in a wide range of aquatic and semi-aquatic breeding sites that are rich in microbial decay. We present an account of the biology and important details of the life cycle of the pupal parasitoid R. quadrispinus in both natural and controlled conditions. Host suitability, the influence of host pupal age and age of the female parasitoid on her production of progeny was evaluated in laboratory tests. Our results showed that the percentage of pupae of E. tenax that were parasitized by R. quadrispinus decreased as pupae matured, and the optimal age for successful parasitism ranged from 1 to 3 days. This biological information could be useful in the biological control of anthropophilic saprophagous eristalines that can act as potential mechanical vectors of pathogens on farms.

In predatory ladybirds male embryos in clutches of eggs infected with male-killing bacterial endosymbionts do not hatch and are consumed by female hatchlings. Moreover, it is predicted that infection with male-killing bacteria should be prevalent in a ladybird population if the fitness advantage to female ladybirds due to the resulting reallocation of resources is high. We compared male-killer bacterial prevalence in two sibling species of the ladybird genus Harmonia that use different host ranges. Harmonia yedoensis feeds mostly on the highly elusive pine aphid and its hatchlings can greatly enhance their ability to capture prey and thus their survival by consuming un-hatched eggs in the clutch. In contrast, Harmonia axyridis feeds on a wide range of prey and consumption of un-hatched eggs by the hatchlings does not necessarily increase their foraging success. In the study area where these two species of ladybird occur sympatrically 14 of the 22 females of H. yedoensis (64%) laid clutches of eggs of which approximately only a half hatched and did not produce male offspring, whereas only one of 12 of the females of H. axyridis (8%) exhibited symptoms indicating infection with male-killing bacteria. In the H. yedoensis that exhibited symptoms of being infected with male-killing bacteria, the total allocation of resources to each hatchling, which was provided by laying larger eggs and eating the un-hatched eggs in the clutch, was considerable. These results suggest that the difference in the prevalence of infection with male-killing bacteria in these two species of ladybird may reflect differences in the quality of their prey and their foraging behaviour.

Larvae of the endangered Scarce Large Blue butterfly, Phengaris (Maculinea) teleius, are initially endophytic, feeding upon developing seeds of Sanguisorba officinalis, but complete their development as social parasites preying on the brood of Myrmica ants. Females show very specific preferences, laying eggs exclusively in young flower heads of the food plant. In the present contribution we report the results of a detailed study of female behaviour at three sites differing in size and abundance of S. officinalis. Tracked individuals switched between feeding on nectar (nectaring), flight and oviposition. They fed exclusively on flowers of reddish, pink or violet colouration, favouring Viccia cracca and S. officinalis. However, females spent about one fifth of the time they were active ovipositing and laid about 20 eggs per hour. Oviposition visits to flower heads were highly stereotyped and followed a specific ritual. Examination of accepted, rejected and randomly sampled flower heads showed that females avoided those containing conspecific eggs and at a site where this was difficult due to a shortage of suitable flower heads, they tended to distribute their eggs evenly among the available flower heads. Our results indirectly indicate the existence of an oviposition-deterring pheromone (ODP) in P. teleius. This hypothetical marking ensures a more even distribution of eggs and possibly increases the chances of survival of caterpillars by reducing competition during both the phytophagous and myrmecophagous stages of development. There are few reports of butterflies producing ODPs and to our knowledge this is the first evidence of their presence in the family Lycaenidae.

Glutamine synthetase (GS) is an essential detoxification enzyme that plays an important role in stress responses; however, little information regarding the function of this enzyme in hymenopteran insects is available. In the present study, we isolated and characterized the gene encoding GS in the Asiatic honeybee, Apis cerana cerana. Multiple alignments and a phylogenetic analysis of GS sequences showed that AccGS belongs to the GSII superfamily and clusters with invertebrate GSs. Real-time quantitative PCR data demonstrated that AccGS is expressed at all developmental stages and in all tissues, with the highest expression observed in the sixth larval instar and in the brain. Moreover, AccGS expression is highly regulated by environmental stress, including xenobiotic, temperature, and ultraviolet light stresses. A disc diffusion assay showed that the recombinant AccGS protein confers resistance to mercuric chloride (HgCl₂) stress in E. coli. This suggests that AccGS may play multiple roles in early development and in environmental stress responses.

Pollen analysis of crop samples was used to identify the food sources of species of beetles of the family Oedemeridae in Bulgaria. A total of 21 species and subspecies were examined; pollen was present in the crops of 18 of the species. Our study is the first to provide information on the food sources of four species and subspecies: Chrysanthia varipes varipes, Anogcodes seladonius turcicus, A. ruficollis and Oedemera flavicans. New data on food sources for many of the other species studied was obtained and feeding on pollen of gymnosperms and honeydew by some of the species was confirmed. Possible mechanisms for digesting pollen are discussed. There was no evidence of either the pollen germinating or being predigested in the crop of the beetles.

Body size is one of the most investigated traits in studies of sexual selection in fruit flies of the genus Drosophila. In D. melanogaster, size has often been correlated with male mating success, i.e. larger males were more successful in achieving copulations both in laboratory and field conditions. In the present paper, we investigated if male body size is a sexually selected trait in competitive conditions, when full-sibs that developed at two different temperatures (18 and 25°C) competed for females. Males developed at a lower temperature were significantly larger than those reared at a higher temperature, but they were not more successful in mating. We conclude that when body size is significantly induced by temperature variability, it is not correlated with male mating success.

Invertebrate diversity has rapidly declined throughout Europe during the last century. Various reasons for this decrease have been proposed including human induced factors like climate change. Temperature changes alter distributions and occurrences of butterflies by determining habitat conditions at different scales. We evaluated changes in the composition of butterfly communities recorded at nine areas of fallow ground in south-western Germany in 1973, 1986, 2010 and 2012 using Pollard's transect technique. To demonstrate the importance of climatic changes in affecting butterfly communities, we calculated the community temperature index (CTI) for each butterfly community in each year. Although they increased slightly, the CTI-values did not match the temperature trends recorded in the study region. However, the reduction in the standard deviations of the CTIs over time is reflected in the marked loss of cold- and warm-adapted species due to their inability to cope with temperature and land-use induced habitat changes. Results of our butterfly surveys indicate a marked decline in species richness and striking changes in the composition of the butterfly communities studied. This trend was most pronounced for habitat specialists, thus mirroring a depletion in trait diversity. Our results indicate that, in the course of large-scale anthropogenic changes, habitat degradation at smaller scales will continuously lead to the replacement of habitat specialists by ubiquitous species.

Gronotoma micromorpha is a solitary egg-pupal and larval-pupal parasitoid. In a previous study, the developmental time of this parasitoid decreased with the age of its leaf miner host, Liriomyza trifolii, being significantly longer when the host was 0-day old (= 0-24 h old egg) than when 4 days old (mature larva) at the time of parasitization at 25°C. In the present study, the suitability and acceptability of 0 and 4 day old L. trifolii as hosts for G. micromorpha, and the ovipositional preference of this parasitoid for these two developmental stages of the host were examined. No significant difference was found in the size of the parasitoid offspring that emerged from hosts parasitized at these two developmental stages. There was no significant difference in the acceptability of the two developmental stages of L. trifolii in no choice tests, but 4 day old larvae were preferred to 0 day old eggs in choice tests. Moreover, parasitization of eggs by G. micromorpha did not appear to result in more of the eggs dying before they hatched. Non-reproductive killing (host feeding and host stinging without oviposition) of host eggs by this parasitoid was also not detected.

Six microsatellite markers were developed for the larch budmoth Zeiraphera diniana Guénée 1845, using two enrichment protocols. The number of alleles ranged from 3 to 15 per locus and observed heterozygosities ranged from 0.09 to 0.98 for the 69 individuals genotyped. Using these markers significant genetic differentiation between one population from Poland and samples from Alpine populations in France and Switzerland (overall F_ST = 0.0298) was detected. However, the two Alpine samples did not differ significantly. These microsatellite markers are valuable tools for studying the population genetics of Zeiraphera diniana.

Studies on the effect of varying light intensity on the spatial distribution of flying insect communities are rare, particularly in complex ecosystems like forests. The horizontal and vertical distribution of Heteroptera was studied at different scales in a large deciduous forest area, the "Steigerwald", in southern Germany. Diversity was affected by (1) vertical position: it was significantly higher near the ground than in the canopy of beech-dominated forests but similar in oak-dominated forests; within the canopy of beech-dominated forests, diversity was significantly higher in the upper than in the lower canopy of intermixed oak trees but similar in beech trees; (2) canopy cover, but in oak forests the response depended on the vertical position: increasing significantly close to the forest floor with decreasing canopy cover, but showing an opposite trend in the canopy; so that in sparse stands (little canopy cover) diversity was significantly higher near the ground, whereas where the forest canopy was medium or dense diversity was higher in the canopy. Moreover, community composition of Heteroptera near the ground differed from that in the canopy in both forest types and near the ground between stands in oak-dominated forest that had canopies of different densities. Results clearly indicate that light intensity is an important direct or indirect factor structuring Heteroptera communities. While in the canopy differences in leaf quality and microhabitats might be important, near the forest floor it is more likely to be the diversity of herbaceous plants.

Theoretical models predict that brood guarding may evolve in situations where eggs are costly to produce or when handling times are long. This study reveals that females of the secondary hyperparasitoid Trichomalopsis apanteloctena guarded cocoon broods of Cotesia kariyai, a gregarious endoparasitoid. Hyperparasitoid females also monopolized host resources and protected their offspring by driving away other conspecific hyperparasitoid females. The females exhibited antagonistic behavior towards competitors through threatening body postures, biting and chasing. Using a video camera to determine how long a hyperparasitoid female attended and parasitized cocoons within a single host brood, it was found that after about 4 days, cocoon guarding behavior became much less apparent. Moreover, more than 90% of hosts were typically parasitized by a hyperparasitoid female over the course of 4 days after she commenced brood guarding. Observations of egg production during a female's lifetime revealed a physiological interval rhythm that typically lasted 3-4 days, which correlates almost exactly with the period during which the cocoons were guarded. To confirm the giving-up time for a host cocoon brood, hyperparasitoid females were given access to 24 h-old cocoon clusters, each containing 60-100 individual cocoons. Ninety percent of the females remained on cocoons for approximately 72 h. Furthermore, twenty-five percent of wasps continued attending and presumably guarding host cocoon broods for more than 138 h after the female first attended the brood. C. kariyai larvae pupate within a few hours of egression from their host and emerge as adults about 5 days (120 h) later. Therefore, many hyperparasitoid females continued to guard older host cocoons of greatly reduced quality as a resource for their progeny and some even after eclosion of the primary parasitoid. Late-brood guarding enabled a hyperparasitoid female to protect her own progeny from other hyperparasitoid females that readily attacked and killed them when she was removed. Our study thus reveals that extended guarding behavior is an adaptive mechanism that probably plays an important role in the survival of the original brood.

Griffiths G.C.D.: Anthomyiidae. In Griffiths G.C.D. (ed.): FLIES OF THE NEARCTIC REGION. Vol. VIII, Part 2, No. 15. pp. 2485-2635. E. Schweizerbart'sche Verlagsbuchhandlung, Stuttgart, 2004, 151 pp., 178 Figs. ISBN 3-510-70027-9.

New aspects of the life-cycle of the flightless wing-polymorphic bug Pyrrhocoris apterus (L.) are described. It was found that 1.1-14.9% of the adult females of P. apterus in 11 samples collected from 4 populations in the South Bohemia had mated prior to entering diapause and their ovaries were in the pre-vitellogenic stage of development with viable sperm in their spermathecae. Some of these females successfully overwintered. The percentage of overwintering females that had been fertilized ranged from 1.1 to 7.2 % and depended on the month and year of collection. The results indicate that pre-diapause mating and overwintering of fertilized females is not unusual in populations of P. apterus in the Czech Republic and and that sperm in overwintering female survives for at least 7-8 months.

The prevalence of obligate parasitic fungi may depend partly on the environmental conditions prevailing in the habitats of their hosts. Ectoparasitic fungi of the order Laboulbeniales (Ascomycetes) infect arthropods and form thalli on the host's body surface. Although several studies report the incidence of infection of certain host species by these fungi, quantitative data on laboulbenialean fungus-host arthropod interactions at the host assemblage level are rarely reported. To clarify the effects of host habitats on infection by ectoparasitic fungi, the incidence of infection by fungi of the genus Laboulbenia (Laboulbeniales) of overwintering carabid beetles (Coleoptera: Carabidae) in three habitats, a riverside (reeds and vines), a secondary forest and farmland (rice and vegetable fields), were compared in central Japan. Of the 531 adults of 53 carabid species (nine subfamilies) collected in the three habitats, a Laboulbenia infection of one, five and one species of the carabid subfamilies Pterostichinae, Harpalinae and Callistinae, respectively, was detected. Three species of fungus were identified: L. coneglanensis, L. pseudomasei and L. fasciculate. The incidence of infection by Laboulbenia was higher in the riverside habitat (8.97% of individuals; 14/156) than in the forest (0.93%; 2/214) and farmland (0%; 0/161) habitats. Furthermore, the incidence of infection by Laboulbenia in the riverside habitat ranged from 0 to 33.3% and differed significantly in the ten microhabitats (riverbank, edge of track, tall reeds, kudzu vines, slope of a hollow, rotten wood, vine reeds, under stones, the shoulder of a terrace and marshy ground) where the carabid beetles overwintered. These results suggest that host habitats and microhabitats are closely associated with successful infection by laboulbenialean fungi.

We present a molecular phylogeny incorporating all five species of the Palaearctic geometrid genus Lythria, based on a 2810-bp combined data matrix comprising the full sequence of the mitochondrial gene cytochrome oxidase subunit one (COI) and fragments of the nuclear genes elongation factor 1 alpha (EF-1α) and wingless (wgl). L. venustata, which was recently rediscovered from Kazakhstan, is shown to be sister taxon to all other members of the genus. The remaining species within the genus form two pairs of sister species: L. purpuraria groups together with L. plumularia, and L. cruentaria with L. sanguinaria. The phylogeny is well supported by characters of the male genitalia of all Lythria species. In addition to the molecular phylogeny of the genus Lythria, we illustrate the external appearance of L. venustata for the first time and describe the anatomy of its male genitalia.

To determine the relationship between protein expression and insect diapause, a proteomic approach was used to investigate the proteins extracted from larvae of the wheat blossom midge Sitodiplosis mosellana Gehin at different developmental stages, including pre-diapause, over-summering diapause, over-wintering diapause and post-diapause. Using 2-DE gels stained with coomassie brilliant blue, about 300 protein spots were detected in the extracts of pre-diapause larvae and 275 for those in each of the other stages. There were 91, 92 and 95 protein spots that showed more than a 2-fold change in abundance in the over-summering diapause, over-wintering diapause and post-diapause stages compared with pre-diapause. Eight protein spots, which showed the greatest difference in the larvae at different stages of diapause, were analyzed using Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Seven of them were successfully identified from their peptide mass fingerprints using the NCBInr database. They were proopiomelanocortin, NADH dehydrogenase subunit 1 and F10F2.5, which were up-regulated or unique to pre-diapause larvae, IKK interacting protein isoform 2 up-regulated in diapause and post-diapause larvae, GA10647-PA unique in over-wintering diapause larvae, purple CG16784-PB isoform B and B0228.6 up-regulated in over-summering and over-wintering diapause larvae. The potential functions of these proteins during wheat blossom midge diapause are discussed.