Isolation and characterization of highly polymorphic microsatellite loci in the 2‐Spot Ladybird,Adalia bipunctata

Contrary to theoretical predictions, female 2‐spot ladybirds (Adalia bipunctata) mate many more times than necessary to maintain high fertilisation success and may gain through the acquisition of material or genetic benefits. In order to investigate this mating system in detail, microsatellite markers have been isolated using a modified enrichment technique. Thirty‐nine loci were successfully amplified by polymerase chain reaction (PCR), of which only two were monomorphic. Detailed characterization of ten loci revealed very high levels of polymorphism. These markers are likely to be invaluable tools with which to study population genetics and patterns of paternity in this species.     

Glutathione S-transferase isoenzymes in the two-spot ladybird, Adalia bipunctata (Coleoptera: Coccinellidae)

Isoenzymes of glutathione S-transferase (GST) in adult Adalia bipunctata, an aphidophagous predator, were studied. Cytosolic GST activity was studied in each beetle developmental stage. The highest activities towards both 1-chloro-2,4-dinitrobenzene (CDNB) and 2,4-dinitro-1-iodobenzene (DNIB) occurred in adults. The enzyme distribution was investigated in adults. While most of the enzymatic activity was found in the abdomen (40-50 and 34-63% respectively) using several concentrations of both CDNB and DNIB, significant differences were observed for the head and the thorax depending on the substrate. Activities were more abundant in the thorax with DNIB (37-47%) compared to the 13-19% obtained with CDNB. Some GST activity was also detected in the elytra. GSTs were purified by epoxy-activated Sepharose 6B affinity chromatography and applied to an HPLC column to determine the native molecular weight (69 kDa). Three isoenzymes were separated by chromatofocusing at pH ranges 7-4. Three bands with molecular mass from 23 to 26 kDa were visualised on SDS-PAGE. Their isoelectric points were 6.66, 6.36, and 6.21. The substrate specificities and the kinetic parameters (Vm and Km) of the isoenzymes showed large differences depending on the isoenzyme. Arch.     

The inheritance of a wingless character in the 2spot ladybird (Adalia bipunctata)

A viable wingless 2spot ladybirdAdalia bipunctata (L.) was found in the wild. Breeding through four generations revealed that the wingless trait was controlled by a recessive allele which displays variable levels of expression. The wingless ladybird is discussed in relation to its potential as a biocontrol agent. One ladybird also occurred in this stock which is suggestive of a supergene controlling the colour polymorphism in this species.     

Expansion of the geographical distribution of an exotic ladybird beetle, Adalia bipunctata (Coleoptera: Coccinellidae), and its interspecific relationships with native ladybird beetles in Japan

The first record of the exotic ladybird beetle, Adalia bipunctata (Coleoptera; Coccinellidae), in Japan was in 1993 at Osaka Nanko Central Park. Since that time, studies on the life history and geographical distribution of A. bipunctata have been ongoing, and its establishment in the Osaka Nanko area has been confirmed. A. bipunctata is a predacious ladybird beetle and a member of a guild that overlaps in habitat and prey with that of native ladybird beetles such as Harmonia axyridis and Menochilas sexmaculatus. We investigated the distribution of A. bipunctata and its interspecific relationships with native predacious ladybird beetles. In some areas, A. bipunctata was dominant in interspecific relationships with native ladybird beetles. For the first 10 years after A. bipunctata was discovered, it occurred only in the Osaka Nanko area, but the present geographical distribution indicates that it has expanded its range. Though the population density of this species was highest at the area recorded first, and tended to decrease in inverse proportion to the distance from Osaka Nanko Central Park, a satellite occurrence was observed in a remote area. The numbers of aphid and tree species (leaf shelter for aestivation and over-wintering) utilized by A. bipunctata have recently increased. Such increases will cause the rate of distribution of A. bipunctata to accelerate. Interspecific competition between H. axyridis and A. bipunctata, which occurs earlier than H. axyridis, may be avoided by desynchronization of the occurrence seasons, and another common predacious ladybird beetle, Coccinella septempunctata, may escape interspecific competition by habitat segregation.     

Dimorphism of wings, wing-folding and wing-toiletry devices in the ladybird, Rhyzobius litura (F.) (Goleoptera: Goccinellidae), with a discussion of inter-population variation in this and other wing-dimorphic beetle species

Dimorphism of tergal wing-folding spicule patches and wing-toiletry fringes accompanies wing dimorphism in the ladybird, Rhyzobius litura. Such dimorphism of 'ancillary' structures is of rare occurrence in the Coleoptera; other instances are discussed. Data are presented concerning the incidence of long and short winged individuals of R. litura in samples from six populations in southern England. Factors likely to influence the relative proportions of the morphs in small populations, and in samples of different kinds, are discussed. Evidence that inter-population variation in the incidence of macropterous individuals has a 'geographical' component in R. litura is inconclusive.     

Differences between the 7-spot and 2-spot ladybird beetles (Coccinellidae) in their toxic effects on a bird predator

Abstract. 1. Experiments with nestling blue tits Parus caeruleus L. examined the effects of feeding them 7-spot ladybird Coccinella septempunctata (L.), 2-spot ladybird Adalia bipunctata (L.) or controls Tenebrio sp.
2. A feeding rate of 51/3 7-spot ladybirds per day 'killed' nestlings within 2 days. Three of six nestlings fed at half this rate survived 4 days. In contrast, all experimental birds survived sixty-four 2-spot ladybirds fed over 4 days (equivalent by weight to the high 7-spot diet).
3. Nestlings fed 7-spot ladybirds also grew more slowly than birds fed 2-spot ladybirds. The latter birds showed no difference in weight gain from the controls although they begged more strongly for food.
4. Cadavers of birds fed 7-spot ladybirds showed evidence of severe liver damage.
5. The results indicate a substantial difference between the ladybird species in their toxicity to nestling blue tits. They support the hypothesis that 2-spot ladybirds are largely-edible, polymorphic Batesian mimics of well-protected, monomorphic species such as the 7-spot ladybird.     

Bacteria in ovarioles of females from maleless families of ladybird beetles Adalia bipunctata L. (Coleoptera: Coccinellidae) naturally infected with Rickettsia,Wolbachia, and Spiroplasma

Ovarioles were found to be infected with Spiroplasma, Wolbachia, and Rickettsia in Adalia bipunctata females with maleless progeny in different natural populations. Ooplasm was infected with few Wolbachia bacteria. In ooplasm infected by Rickettsia, bacteria were present in small foci. Spiroplasmas were found encapsulated into ooplasm from the wider intercellular spaces between epithelial and oocyte cells. The cytoplasm of follicular epithelia infected with Rickettsia was heavily destroyed, but the nucleus was intact and free from bacteria. The essential feature of follicular epithelium cells from Spiroplasma and Wolbachia infected A. bipunctata females was inclusions of three types: crystalline, filaments, and concentric myelin-like lamellae. Observations of smears prepared from ovaries of A. bipunctata from natural populations revealed a low concentration of bacteria within a microscopy field (less 10 bacteria) in more than 90% of specimens, and only a few ovaries were heavily infected. Two different ways of bacterial invasion of the oocyte are suggested: Spiroplasma-like, through the intercellular spaces in the epithelium and Rickettsia-like, through the cytoplasm of follicular epithelium cells. Bacteria were not found in germarium zones and we suggest that each follicle is infected from haemolymph.     

A countrywide survey of colour morph frequencies in the spider Enoplognatha ovata (Glerck) (Araneae: Theridiidae): evidence for natural selection

Data on the frequencies of the main colour morphs ( lineata, redimita and ovata ) of Enoplognatha ovata have been collected from a total of 454 Ordnance Survey 10 km squares distributed throughout Great Britain. Only c. 0.5% of samples were monomorphic, for lineata in each case. Multiple regression analysis has been used to assess possible associations between morph frequencies and principal components derived from nine environmental variables. The distribution of morph frequencies is not random but shows weak clines associated with certain climatic factors. These large scale clines indicate the action of natural selection although very local variations in morph frequencies may result from selection and/or drift.     

Home-range characteristics and the influence of seasonality on female reproduction in white-handed gibbons (Hylobates lar) at Khao Yai National Park, Thailand

A three-year (2001-2003) study was carried out on the home range characteristics of seven wild white-handed gibbon (Hylobates lar) groups focusing on the spatio-temporal distribution of food resources at Khao Yai National Park in northeastern Thailand. These results were combined with 23 years (1980-2003) of reproductive performance data on seven females from the same focal groups. Reproductive performance was equal among females with regard to birth, weaning and maturation ratios, and independent of variation in food availability. Offspring mortality, however, was significantly positively correlated with home-range size. In addition, there was an increase in offspring mortality just after weaning, suggesting that the increase in the daily distance traveled by juveniles contributed to this mortality. Conceptions clustered during the first half of the year when food production was at its peak, which presumably allowed females to accumulate sufficient body reserves to resume ovarian cycling. Our results place Khao Yai gibbons closer to Cercopithecidae than great apes in terms of the temporal pattern of reproductive events, though gestation, lactation, inter-birth interval, and offspring maturation are considerably longer in gibbons, placing them closer to the other apes. Our findings underline the unique phylogenetic position of these small-bodied apes in terms of reproductive patterns in primates.     

Natural selection on a leaf-shape polymorphism in the ivyleaf morning glory (Ipomoea hederacea)

Leaf shape is one of the most variable plant traits. Previous work has provided much indirect evidence that leaf-shape variation is adaptive and that leaf shape influences thermoregulation, water balance, and resistance to natural enemies. Nevertheless, there is little direct evidence that leaf shape actually affects plant fitness. In this study, we first demonstrate that populations of the ivyleaf morning glory, Ipomoea hederacea, in North and South Carolina are frequently polymorphic at a locus that influences leaf shape. We then employ several field experiments to show that this polymorphism is subject to selection. In two of the experiments, at different sites, heterozygotes enjoyed a fitness advantage over both homozygotes. At a third site, in one year directional selection favored lobed leaves, whereas in a second year the pattern of fitnesses was consistent with similar directional selection or heterozygote superiority. Computer simulations of heterozygote advantage under the high selfing rates of I. hederacea indicate that balancing selection of the magnitude observed can by itself stabilize the polymorphism, although spatially and temporally variable selection may also contribute to its long-term maintenance.     

Heterosis, epistasis and linkage disequilibrium in a wild population of the polymorphic butterfly Danaus chrysippus (L.)

The colour polymorphism of the Danaus chrysippus population at Dar es Salaam, East Africa, is controlled at three major loci, each with two alleles. Two of the loci, one governing ground colour and the other forewing pattern, are closely linked. The third locus, determining hindwing pattern, assorts independently. Thirty-eight broods raised from wild mated pairs, Fl and F2 generations gave 857 offspring of 23 genotypes (out of 27 possible). The forewing length, taken as an index of size, was investigated in relation to the genotype. Heterosis is evident at all three loci. The two linked loci show epistatic interaction of an unexpected kind: double heterozygotes are smaller than heterozygotes at only one locus but larger than double homozygotes. The heterotic effect at the third, unlinked locus is the most pronounced and is additive to that at the other two. Heterosis is more marked in males than females. The possibility that body size has importance in connexion with sexual selection, food resources and mimetic relationships is discussed. Analysis of gene and chromosome frequencies in the wild parents of 61 broods suggests that double heterozygotes for the two linked loci may have heterozygous advantage. Seventy-eight per cent of chromosomes are repulsion phase: thus, there is pronounced linkage disequilibrium which must be maintained by selection as crossing over is almost 296. In particular, the chromosome carrying both dominant alleles in coupling is rare. Consideration of the centres of distribution and present ranges of the alleles at all three loci suggests that three geographical races, aegyptius, dorippus and alcippus, were isolated by forest barriers, during wet periods in the Pleistocene, in south-west, north-east and north-west Africa respectively. They have probably expanded their ranges in the post-glacial period to overlap and interbreed in central and east Africa. Either heterozygous advantage or seasonal (directional) selection or a combination of both is responsible for the persistence of the polymorphism.     

Activity and shelter selection by Myotis myotis and Rhinolophus hipposideros hibernating in the Kateřinská cave (Czech Republic)

In 1992–1993, the bat species Myotis myotis and Rhinolophus hipposideros hibernating in the Kateřinská cave were investigated by means of regular censuses without any handling and marking of the animals. Three basic parameters of their shelters were recorded (position in cave, type and relative height). In total, during 26 checks we registered 1141 findings of nine bat species. Movement activity, expressed as percentage of new findings during a particular visit, was registered during the whole winter season. Its level fluctuated in different ways and the hibernation period of R. hipposideros could be divided into three different parts, while the level of M. myotis movement activity was relatively high during all season. The shelter selection of R. hipposideros was not dependent on the part of cave where the bats were hibernating, and it did not change during the season. Hibernating specimens of R. hipposideros most frequently used exposed places, in which they were always hanging free. Myotis myotis was registered in all types of shelter with one exception. Rhinolophus hipposideros used mainly the middle part of the cave at a distance between 121 and 180 m from the entrance. The most preferred part of the cave by M. myotis was a small segment of Corridor (between 21 and 30 m), i.e., the entrance part of the cave. Rhinolophus hipposideros is a highly specialized species which prefers parts of the cave with very stable microclimatic conditions and, on the contrary, M. myotis appears to be indifferent to all parameters studied, and it uses the shelters indiscriminately.     

Age, growth and mortality of red bandfish, Cepola macrophthalma (L.), in the western Aegean Sea (Greece)

Age, growth and mortality of red bandfish, Cepola macrophrhalma (L.), from the western Aegean Sea were studied using the otoliths from 1113 fish and the length and weight of 3351 fish. Two regions were established (north and south of the Euripos Strait). The red bandfish grows allometrically (slope of length-weight regressions ≤2) and relatively rapidly until age 4 or 5 years. It is proposed that this rapid linear growth represents an adaptation which evolved to reduce the vulnerability of red bandfish to whole-fish swallowers. Back-calculated lengths-at-age showed significant differences between regions and between sexes. Males were larger than females after age 2 years. Maximum age of northern fish was also higher than southern fish (8 and 5 years respectively). Southern fish grow at a faster rate (K=0.379) but to a significantly smaller size (L_x=424.8mm t.l. , W_x=25.1 g) than northern fish (K=0.214, L_x=676.1 mm t.l. , W_x=90.9 g). Mortality rates (total and natural instantaneous mortalities) of southern fish were also higher than northern fish. We suggest that these variations represent responses to the different conditions of temperature and food availability prevailing in the two regions.     

Surviving winter: diapause syndrome in the southern green stink bug Nezara viridula in the laboratory,in the field,and under climate change conditions

The southern green stink bug Nezara viridula (L.) (Heteroptera: Pentatomidae) has long attracted attention not only as a serious pest of numerous agricultural crops, but also as a species expanding its range in many parts of the world. Nezara viridula has also been widely used as a model in different experimental studies. The present review focuses on reproductive (i.e. adult) winter diapause, which is the pivotal element of the species' seasonal cycle. Results from numerous field experiments and observations, as well as laboratory ecophysiological investigations conducted during the few last decades, are analyzed and interpreted. Experimental findings are used to describe in detail the dynamics of physiological changes during overwintering. Reproductive diapause in N. viridula is controlled in both sexes by photoperiodic conditions. The induction of diapause is associated with a reversible change of body colour from green or yellow to russet (or brown). The proper timing of adult emergence and the induction of diapause, as well as the size of adults, is vitally important for successful overwintering. Nezara viridula has been shown to respond strongly to the current trend in climate change by shifting the limit of its northern range, particularly in central Japan. Analysis of historic climate data suggests that the environmental conditions during the last few decades have become more favourable for the overwintering survival of N. viridula in many locations in central Japan. This has likely promoted the northward spread of the species. The relationships between reproductive diapause, reversible body colour change, overwintering success and the recent range expansion are analyzed. Perspectives of the range dynamics of the species are discussed in light of further predicted climate change.     

Linkage disequilibrium and natural selection for mimicry in the Batesian mimic Hypolimnas misippus (L.) (Lepidoptera: Nymphalidae) in the Afrotropics

On two occasions, on opposite sides of the African continent (Cape Coast, Ghana, and Dar es Salaam, Tanzania), high adult population densities in the polymorphic butterfly Hypolimnas misippus (a presumed mimic of Danaus chrysippus) were followed by linkage disequilibrium in combinations of fore‐ and hindwing colour patterns. On both occasions, disequilibrium was caused by significant changes in morph frequencies favouring rarer and more mimetic forms. Recaptures were too few for analysis at Dar, although the changes there took place within a single generation and must have been the result of differential survival. Recapture rate data and survival rate estimates at Cape Coast support the hypothesis that selective predation was responsible, as does the observation of synchronous linkage disequilibrium at Dar in the model D. chrysippus, indicating parasitic mimicry. There was clear selection for the perfection of mimicry for forewings at Dar and for hindwings at Cape Coast. Disequilibrium is also reported for two other sites, Legon (Ghana) and Boksburg (South Africa) and, in all four sites, it was associated with an increase in the most mimetic forms. New chemical evidence is presented to support the contention that D. chrysippus is a defended model. Although all the evidence leads to the conclusion that H. misippus is a Batesian mimic of D. chrysippus, many questions remain, particularly with regard to the identity of predators, the episodic nature of selective predation events, and their apparent lack of lasting and significant impact on overall gene frequencies. We conclude that H. misippus presents both challenges and opportunities for studies on mimicry, and we suggest that linkage disequilibrium can be a useful generic indicator for Gestalt predation on polymorphic prey. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 180–194.     

Carry‐over effects of conditions at the wintering grounds on breeding plumage signals in a migratory bird: roles of phenotypic plasticity and selection

To understand the consequences of ever‐changing environment on the dynamics of phenotypic traits, distinguishing between selection processes and individual plasticity is crucial. We examined individual consistency/plasticity in several male secondary sexual traits expressed during the breeding season (white wing and forehead patch size, UV reflectance of white wing patch and dorsal melanin coloration) in a migratory pied flycatcher (Ficedula hypoleuca) population over an 11‐year period. Furthermore, we studied carry‐over effects of three environmental variables (NAO, a climatic index; NDVI, a vegetation index; and rainfall) at the wintering grounds (during prebreeding moult) on the expression of these breeding plumage traits of pied flycatcher males at individual and population levels. Whereas NAO correlates negatively with moisture in West Africa, NDVI correlates positively with primary production. Forehead patch size and melanin coloration were highly consistent within individuals among years, whereas the consistency of the other two traits was moderate. Wing patch size decreased with higher NAO and increased with higher rainfall and NDVI at the individual level. Interestingly, small‐patched males suffered lower survival during high NAO winters than large‐patched males, and vice versa during low NAO winters. These counteracting processes meant that the individual‐level change was masked at the population level where no relationship was found. Our results provide a good example of how variation in the phenotypic composition of a natural population can be a result of both environment‐dependent individual plasticity and short‐term microevolution. Moreover, when plasticity and viability selection operate simultaneously, their impacts on population composition may not be evident.     

Lack of association between winter coat colour and genetic population structure in the Japanese hare,Lepus brachyurus (Lagomorpha: Leporidae)

Seasonal changes in fur colour in some mammalian species have long attracted the attention of biologists, especially in species showing population variation in these seasonal changes. Genetic differences among populations that show differences in seasonal changes in coat colour have been poorly studied. Because the Japanese hare (Lepus brachyurus) has two allopatric morphotypes that show remarkably different coat colours in winter, we examined the population genetic structure of the species using partial sequences of the SRY gene and six autosomal genes: three coat colour‐related genes (ASIP, TYR, and MC1R) and three putatively neutral genes (TSHB, APOB, and SPTBN1). The phylogenetic tree of SRY sequences exhibited two distinct lineages that diverged approsimately 1 Mya. Although the two lineages exhibited a clear allopatric distribution, it was not consistent with the distribution of morphotypes. In addition, six nuclear gene sequences failed to reveal genetic differences between morphotypes. Population network trees for 11 expedient populations divided the populations into four groups. Genetic structure analysis revealed an admixture of four genetic clusters in L. brachyurus, two of which showed large genetic differences. Our results suggest ancient vicariance in L. brachyurus, and we detected no genetic differences between the two morphotypes. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 761–776.     

Phenotypic plasticity, seasonal climate and the population biology of Bicyclus butterflies (Satyridae) in Malawi

Abstract.

• 1 Seasonal polyphenism is studied in a community of five African butterflies of the genus Bicyclus at the transition between a wet and a dry season from May to July.
• 2 Butterflies characterized by large eyespots and, especially in B.sufitza (Hewitson), a pale band (the wet season form) are replaced over this period by butterflies lacking conspicuous wing markings (the dry season form, dsf). The latter butterflies also tend to be larger, but more variable in size. Butterflies of an intermediate phenotype are recruited over a comparatively short interim period.
• 3 This turnover coincides with a period of declining temperature and drying of the habitat, including the grasses on which larvae feed. Butterflies are progressively more likely to rest on brown leaf litter rather than on green herbage.
• 4 A relationship with temperature is supported by laboratory experiments with B.saJitza and B.anynana (Butler) showing that increasingly extreme dsf butterflies develop with decreasing rearing temperature in the final larval instar.
• 5 Some differences in behaviour and activity were observed between the seasonal forms. Butterflies of the dsf develop ovarian dormancy and fat bodies. They can survive to reproduce at the beginning of the rains in November.
• 6 Capture-recapture experiments showed that the adult butterflies have a comparatively long life expectancy and are quite sedentary.
• 7 The results are discussed in relation to a hypothesis linking the polyphenism to seasonal changes in resting background and selection for crypsis.

     

Foraging area and hunting technique selection of Common Kestrel (Falco tinnunculus) in winter: the role of perch sites

In winter, Common Kestrel minimizes energy expenditure by using the low-cost, low-profit technique of perch hunting. Existence of the perch sites is the precondition of perch hunting. Therefore, one can predict that the kestrels would prefer a habitat with more perch sites, and perch sites should have an important role in the kestrels' hunting technique use, habitat selection and habitat use in winter. To test this prediction, the authors manipulated two areas in a grassland. They increased the potential perch sites in one area with bamboo poles (hereafter test area) and kept another as control (hereafter control area). They observed and compared the kestrels' use and their behaviors in these two areas. Far more kestrels appearing in the test area with increasing perch sites than in the control area were recorded. The kestrels stayed in the test area with more perch sites significantly longer than in the control area. And in the test area with more perch sites, kestrels hunted 77.24% of the total hunting with the technique of perch hunting. In the control area, kestrels hunted only with the technique of flight-hunting. There was a significant correlation between the technique used by kestrels and the areas with or without perch sites. In the test area with increasing perch sites kestrels spent 51.8% of their time in perching and 12.1% in air, which were 30.1% and 34.8%, respectively, in the control area. There was no significant difference in hunting profit between areas. The results suggest that perch sites play an important role in the selection of hunting technique and foraging habitat for kestrels in winter, and kestrels appear to prefer the habitat with suitable perch sites in winter.     

栖木在越冬红隼（Falco tinnunculus）的觅食地与捕食方式选择中的作用

栖木是重要的生境因子,对于鸟类生境选择和利用具有重要意义。2003年9月-2004年3月,采用固定样地对照观测的方法,分析研究栖木在越冬红隼（Falco tinnunculus）的栖息地选择与捕食方式选择中的作用。研究结果表明,增加了栖木的试验区（下称试验区）,红隼的出现频率远大于未增加栖木的对照区（下称对照区）;试验区红隼个体平均停留时间显著长于对照区;在试验区红隼栖停捕食占捕食次数的77.24%,而在对照区红隼只进行飞行捕食,红隼捕食方式和样地中是否增加栖木具有极显著相关性;试验区,红隼栖停行为时间占51.8%,大于对照区（30.1%）,对照区中,红隼在空中的行为如飞行、翱翔、悬停等行为时间所占比例为34.8%,远高于试验区（12.1%）。这些结果表明,栖木在越冬红隼的栖息地选择和捕食方式选择中具有重要的作用,栖木的存在是栖停捕食的先决条件,冬季红隼偏好于具有较好栖木条件的栖息地,并且偏向于采取低消耗低收益的栖停捕食。