Number Of Ovarioles Research Articles

Associations between reproduction and work in workers of the Asian hive bee Apis cerana

If a honey bee (Apis spp.) colony becomes queenless, about 1/3 of young workers activate their ovaries and produce haploid male-producing eggs. In doing so queenless workers maximize their inclusive fitness because the normal option of vicarious production of relatives via their queen’s eggs is no longer available. But if many workers are engaged in reproduction, how does a queenless colony continue to feed its brood and forage? Here we show that in the Asian hive bee Apis cerana hypopharyngeal gland (HPG) size is larger in queenless workers than in queenright workers and that bees undertaking brood-rearing tasks have larger HPG than same-aged bees that are foraging. In queenless colonies, workers with a smaller number of ovarioles are more likely to have activated ovaries. This reinforces the puzzling observation that a large number of ovarioles reduces reproductive success in queenless A. cerana. It further suggests that reproductive workers either avoid foraging or transition to foraging later in life than non-reproductive workers. Finally, our study also showed that ovary activation and larger-than-average numbers of ovarioles had no statistically detectable influence on foraging specialization for pollen or nectar.

Convergent Reduction of Ovariole Number Associated with Subterranean Life in Beetles.

BackgroundSome species of obligate cavernicolous beetles are known to possess a unique feature—a contraction of the larval cycle. In contrast to many other subterranean beetles, life-cycle contraction in Trechini ground beetles (Carabidae) is correlated with a reduction in the number of eggs and a drastic reduction in the number of ovarioles. This remarkable peculiarity has only been reported for a small number of closely related species.ResultsWe give a description of the female internal reproductive system for six species of Trechini, including five subterranean species, with a particular focus on the western Pyrenean radiation of Aphaenops, a group for which nothing is known regarding the early life stages. We redescribe the internal female genitalia of A. crypticola Linder. Study of the ovarioles allowed us to infer the postembryonic development of the larvae for each species examined. We then used a phylogenetic framework to recognize two independent reductions in the number of ovarioles in the Pyrenean lineage. We discuss the multiple convergent evolutions in ovariole number and the potential link between a reduction of ovariole number and troglobiomorphism in a phylogenetic context.ConclusionsThere is an extreme reduction in ovariole number and size within the species studied; the eggs produced by small ovarioles have a remarkably large size. A reduction to one ovariole has occurred independently at least twice in this subterranean group. A reduction in the number of ovarioles in ground beetles is one of the striking consequences of subterranean specialization and it is correlated with another remarkable adaptation of subterranean beetles, a reduction in the number of larval instars.

Phenotypic Variation and Sexual Size Dimorphism in Dichroplus elongatus (Orthoptera: Acrididae).

Patterns of body size evolution are of particular interest because body size can affect virtually all the physiological and life history traits of an organism. Sexual size dimorphism (SSD), a difference in body size between males and females, is a widespread phenomenon in insects. Much of the variation in SSD is genetically based and likely due to differential selection acting on males and females. The importance of environmental variables and evolutionary processes affecting phenotypeic variation in both sexes may be useful to gain insights into insect ecology and evolution. Dichroplus elongatus Giglio-Tos is a South American grasshopper widely distributed throughout Argentina, Uruguay, most of Chile, and southern Brazil. In this study, we analyzed 122 adult females of D. elongatus collected in eight natural populations from central-east Argentina. Females show large body size variation among the analyzed populations and this variation exhibits a strong relationship with fecundity. Our results have shown that larger females were more fecund than smaller ones. We found that ovariole number varied along a latitudinal gradient, with higher ovariole numbers in populations from warmer locations. A considerable female-biased SSD was detected. SSD for three analyzed morphometric traits scaled isometrically. However, SSD for thorax length displayed a considerable variation across the studied area, indicating a larger relative increase in female size than in male size in warmer environmental conditions.

Drosophila melanogaster larvae make nutritional choices that minimize developmental time

Organisms from slime moulds to humans carefully regulate their macronutrient intake to optimize a wide range of life history characters including survival, stress resistance, and reproductive success. However, life history characters often differ in their response to nutrition, forcing organisms to make foraging decisions while balancing the trade-offs between these effects. To date, we have a limited understanding of how the nutritional environment shapes the relationship between life history characters and foraging decisions. To gain insight into the problem, we used a geometric framework for nutrition to assess how the protein and carbohydrate content of the larval diet affected key life history traits in the fruit fly, Drosophila melanogaster. In no-choice assays, survival from egg to pupae, female and male body size, and ovariole number – a proxy for female fecundity – were maximized at the highest protein to carbohydrate (P:C) ratio (1.5:1). In contrast, development time was minimized at intermediate P:C ratios, around 1:2. Next, we subjected larvae to two-choice tests to determine how they regulated their protein and carbohydrate intake in relation to these life history traits. Our results show that larvae targeted their consumption to P:C ratios that minimized development time. Finally, we examined whether adult females also chose to lay their eggs in the P:C ratios that minimized developmental time. Using a three-choice assay, we found that adult females preferentially laid their eggs in food P:C ratios that were suboptimal for all larval life history traits. Our results demonstrate that D. melanogaster larvae make foraging decisions that trade-off developmental time with body size, ovariole number, and survival. In addition, adult females make oviposition decisions that do not appear to benefit the larvae. We propose that these decisions may reflect the living nature of the larval nutritional environment in rotting fruit. These studies illustrate the interaction between the nutritional environment, life history traits, and foraging choices in D. melanogaster, and lend insight into the ecology of their foraging decisions.

Interspecific hybridization does not affect the level of fluctuating asymmetry (FA) in the Drosophila bipectinata species complex.

The Drosophila bipectinata species complex comprises of four very closely related species namely D. bipectinata, D. parabipectinata, D. malerkotliana and D. pseudoananassae. It was found that irrespective of the evolutionary divergence among the species, FA which is reflective of the developmental precision remains nearly same in four species. During the present study, the level of FA in different morphological traits was studied in interspecific hybrids and compared with that of parental species with the view that it would throw light on the degree of divergence between the parental species. If they have not diverged much, the interspecific hybrids may have a similar FA level, incompatibilities between their genomes being negligible. On the other hand, if there is substantial divergence, the level of FA may be higher due to incompatibility between the genomes of the parental species. The morphological traits taken were sternopleural bristle number and wing length in both males and females and ovariole number and sex-comb tooth number in females and males respectively. However, except in a few cases, we could not detect any significant differences in the level of FA in hybrids as compared to pure species. On the other hand, a number of abnormalities like poor viability, dystrophied ovaries, asymmetrical eyes etc., could be detected in hybrids from crosses involving D. pseudoananassae as one of the parents. Therefore, we conclude that specific developmental pathways are more susceptible to developmental disturbances due to genomic incompatibilities than the large complex system bringing about developmental stability.

FGF signaling supports Drosophila fertility by regulating development of ovarian muscle tissues

The thisbe (ths) gene encodes a Drosophila fibroblast growth factor (FGF), and mutant females are viable but sterile suggesting a link between FGF signaling and fertility. Ovaries exhibit abnormal morphology including lack of epithelial sheaths and muscle tissues that surround ovarioles. Here we investigated how FGF influences Drosophila ovary morphogenesis and identified several roles. Heartless (Htl) FGF receptor was found to be expressed within somatic cells at the larval and pupal stages, and phenotypes were uncovered using RNAi. Differentiation of terminal filament cells was affected, but this effect did not alter the ovariole number. In addition, proliferation of epithelial sheath progenitors, the apical cells, was decreased in both htl and ths mutants, while ectopic expression of the Ths ligand led to these cells' over-proliferation suggesting that FGF signaling supports ovarian muscle sheath formation by controlling apical cell number in the developing gonad. Additionally, live imaging of adult ovaries was used to show that htl RNAi mutants, hypomorphic mutants in which epithelial sheaths are present, exhibit abnormal muscle contractions. Collectively, our results demonstrate that proper formation of ovarian muscle tissues is regulated by FGF signaling in the larval and pupal stages through control of apical cell proliferation and is required to support fertility.

Morphometric Identification of Queens, Workers and Intermediates in In Vitro Reared Honey Bees (Apis mellifera).

In vitro rearing is an important and useful tool for honey bee (Apis mellifera L.) studies. However, it often results in intercastes between queens and workers, which are normally are not seen in hive-reared bees, except when larvae older than three days are grafted for queen rearing. Morphological classification (queen versus worker or intercastes) of bees produced by this method can be subjective and generally depends on size differences. Here, we propose an alternative method for caste classification of female honey bees reared in vitro, based on weight at emergence, ovariole number, spermatheca size and size and shape, and features of the head, mandible and basitarsus. Morphological measurements were made with both traditional morphometric and geometric morphometrics techniques. The classifications were performed by principal component analysis, using naturally developed queens and workers as controls. First, the analysis included all the characters. Subsequently, a new analysis was made without the information about ovariole number and spermatheca size. Geometric morphometrics was less dependent on ovariole number and spermatheca information for caste and intercaste identification. This is useful, since acquiring information concerning these reproductive structures requires time-consuming dissection and they are not accessible when abdomens have been removed for molecular assays or in dried specimens. Additionally, geometric morphometrics divided intercastes into more discrete phenotype subsets. We conclude that morphometric geometrics are superior to traditional morphometrics techniques for identification and classification of honey bee castes and intermediates.

Morphological allometry and intersexuality in horsehair-worm-infected mantids, Hierodula formosana (Mantodea: Mantidae).

Parasitic castration is a strategy used by parasites to minimize damage to the host by consuming its reproductive system, which results in the morphological alteration of the host. We determined that the forewing shape and density of the antennal sensilla of field-collected adult male mantids (Hierodula formosana), infected by horsehair worms (Chordodes formosanus) was partially feminized (intersexuality), and both male and female mantids infected by horsehair worms exhibited allometric changes in their wings and walking legs. In addition, the testes of most infected male adults disappeared or reduced in size, whereas the number of ovarioles in infected female adults was unaffected. The infection mainly influenced the structures related to host reproduction and locomotion, suggesting unbalanced energy exploitation and the reduction of parasitic virulence. In addition, the intersexuality of infected male adults indicated that sexual differentiation in insects, which researchers have considered to be an autonomous process, was influenced by the infection. The similarity of the antennae of infected male adults with those of last-instar female nymphs suggested that parasitic juvenilization may cause such feminization, but the mechanism of parasitic influence on insect sex characteristics should be studied further.

A soluble pyrophosphatase is essential to oogenesis and is required for polyphosphate metabolism in the red flour beetle (Tribolium castaneum).

Polyphosphates have been found in all cell types examined to date and play diverse roles depending on the cell type. In eukaryotic organisms, polyphosphates have been mainly investigated in mammalian cells with few studies on insects. Some studies have demonstrated that a pyrophosphatase regulates polyphosphate metabolism, and most of them were performed on trypanosomatids. Here, we investigated the effects of sPPase gene knocked down in oogenesis and polyphosphate metabolism in the red flour beetle (Tribolium castaneum) A single sPPase gene was identified in insect genome and is maternally provided at the mRNA level and not restricted to any embryonic or extraembryonic region during embryogenesis. After injection of Tc-sPPase dsRNA, female survival was reduced to 15% of the control (dsNeo RNA), and egg laying was completely impaired. The morphological analysis by nuclear DAPI staining of the ovarioles in Tc-sPPase dsRNA-injected females showed that the ovariole number is diminished, degenerated oocytes can be observed, and germarium is reduced. The polyphosphate level was increased in cytoplasmic and nuclear fractions in Tc-sPPase RNAi; Concomitantly, the exopolyphosphatase activity decreased in both fractions. Altogether, these data suggest a role for sPPase in the regulation on polyphosphate metabolism in insects and provide evidence that Tc-sPPase is essential to oogenesis.

Pheromones affecting ovary activation and ovariole loss in the Asian honey bee Apis cerana

The Asian hive bee Apis cerana has similar queen mandibular pheromones (QMP) to the Western honey bee Apismellifera. However the effects of individual QMP components have never been tested to determine their effects on the reproductive physiology of A. cerana workers. We fed one queen equivalent of each of the major components of A. cerana QMP to groups of c.a. 500day-old, caged, workers twice a day until the workers were 10days old. Half of the cages were also provided with 10% royal jelly in the food. Workers were sampled each day and dissected to determine the number of ovarioles and the degree of ovary activation (egg development). In cages treated with 9-carbon fatty acids ovary activation was minimal, whereas the 10-carbon acids suppressed ovary activation very little. Royal jelly enhanced ovary activation, especially in cages treated with 10-carbon acids. The number of ovarioles declined with bee age, but the rate of decline was slowed by the 9-carbon acids in particular. The results show conservation of the composition and function of QMP between A. cerana and A. mellifera and support the hypothesis that QMP is an honest signal of queen fecundity rather than a chemical castrator of workers.

The relationship between ovarioles number and female size in blackflies of the high Andes of Colombia.

Fitness of a single species is often measured in terms of the number of viable offspring produced. We explore the relationship between the number of ovarioles and the size of females in five species of Simuliidae in two páramo regions of Colombia. Individuals of seven species of female blackflies that landed on animals were collected from mules in the Ucumarí Regional Park (RPNU) and from cattle in the Chingaza Natural National Park (ChNNP). The ovarioles of the five most abundant species were dissected out and counted, and a one-factor ANOVA was performed to explore differences in the mean number of ovarioles produced by different female size categories. Simulium ignescens Roubaud and S. ignescens-like species were collected in RPNU and S. ignescens, Simulium muiscorum Bueno et al, Simulium cormonsi Wygodzinsky and Simulium pautense Coscarón & Takaoka, in ChNNP. In addition, we also analyzed the Pearson product-moment correlation between the mean number of ovarioles per blackfly and female size within species using those which more than 20 individuals were collected. Ovarioles were meroistic-polytrophic type. A multiple range test (least significant difference) indicated that the largest size group had the largest mean number of ovarioles per female. In the other size groups, there were approximately equal numbers of ovarioles per female. Pearson's correlation coefficient was separately measured and indicated a 50% correlation between female size and ovariole number in S. ignescens. Although ovariole number is probably a genetically driven feature, other environmental and physiological conditions, including infections, can influence the number of eggs.

Reproductive Tract Histology of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae)

Abstract The bronze bug Thaumastocoris peregrinus Carpintero & Dellapé (Hemiptera: Thaumastocoridae) is a pest of Corymbia and Eucalyptus. Thus far, there are no effective control methods for this pest in commercial plantations. A study of the reproductive tract could provide basic strategies for controlling this insect. T. peregrinus females and males, of different ages, were obtained from mass rearing and analyzed using light microscopy. The male reproductive tract has a pair of testes with three globular follicles isolated by a peritoneal sheath, and two pairs of well-developed mesadenial tubular accessory glands. The female reproductive tract includes a pair of ovaries, each with two meroistic telotrophic ovarioles, opening into two long lateral oviducts, which joints in a short common oviduct. This common oviduct ends in a large and folded, thick-cuticle lined bursa copulatrix. Eggs with embryos in the stages of anatrepsis, catatrepsis, and post dorsal closure were found in the reproductive tract of females. The reproductive tract of T. peregrinus males and females are similar to those of other Thaumastocoridae, but differ in the number of ovarioles per ovary in its females.

The Biology of Senegalese Grasshopper (Oedaleus Senegalensis, Krauss, 1877) (Orthoptera: Acrididae)

The Senegalese grasshopper, Oedaleus senegalensis (Orthoptera: Acrididae) is known as pest of economic importance which cause damage to millet and sorghum crops besides pastures in western Sudan. The study was done to investigate the biology of the insect on its natural habitat. Observations in the field were made twice/ week, from April-November each year for three continuous seasons 2006/2007, 2007/2008 and 2008/2009, in two selected sites one around Gellabiya village in North Kordofan State west Elobeid town. The other was around Kuik village in South Kordofan State north Kadugli town. The nymphs and the adults were collected by sweep net from the field and transferred to the laboratory for study. Female adults of the Senegalese grasshopper were dissected and examined under binocular Microscope (Paralux, Optigue de précision CE 0602397) to investigate the fecundity through ovarioles numbers and function. Duration of each of the developmental stages of the pest was determined. Results showed that the mean number of ovarioles is 36 / ovary and two egg pods were laid /female. The mean fecundity was 44 eggs. The female laid eggs twice during life cycle. Ovariole function average was 76% in the first laying and 36% in the second laying. Mean developmental duration after hatching of O. senegalensis to adult (nymphal instars) was 25 day. The respective nymphal duration from the 1st to the 5th instar was 4, 5, 5, 5, and 6 days, respectively.

Influences of pyriproxyfen on fecundity and reproduction of the Pharaoh ant (Hymenoptera: Formicidae).

The effects of the juvenile hormone analog pyriproxyfen (at concentrations of 0.1, 0.5, and 1%) on egg production, number of ovarioles, and length of oocytes were examined in queens of the Pharaoh ant Monomorium pharaonis (L.). Pyriproxyfen significantly reduced egg production in queens from week 3 onwards. Queens that were exposed to 1% pyriproxyfen stopped producing eggs at week 8. After 8 wk, ovaries were dissected from all queens, and the number of ovarioles and the length of the largest oocytes were recorded. The ovaries of queens in treated colonies were smaller than those in untreated queens, and the number of ovarioles in the ovaries was significantly lower in all pyriproxyfen-treated queens. Queens treated with the highest concentrations of pyriproxyfen tended to have significantly shorter oocytes than untreated queens. Histological studies of the ovaries revealed that pyriproxyfen caused vacuolation in the ovarioles, thickening of the tunica propria, development of small eggs, and underdevelopment of nurse cells and the follicular epithelium. Exposure to pyriproxyfen reduced egg production and induced severe morphological changes in the ovaries of queens, and the effects increased with increased concentration of pyriproxyfen.

Structure of the female reproductive system of the lac insect, Kerria chinensis (Sternorrhyncha, Coccoidea: Kerridae)

AbstractThe ovaries of female lac insects, Kerria chinensis Mahd (Sternorrhyncha: Coccoidea: Kerridae), at the last nymphal stage are composed of several balloon‐like clusters of cystocytes with different sizes. Each cluster consists of several clusters of cystocytes arranging in rosette forms. At the adult stage, the pair of ovaries consists of about 600 ovarioles of the telotrophic‐meroistic type. An unusual feature when considering most scale insects is that the lateral oviducts are highly branched, each with a number of short ovarioles. Each ovariole is subdivided into an anterior trophic chamber (tropharium) containing six or seven large trophocytes and a posterior vitellarium harbouring one oocyte which is connected with the trophic chamber via a nutritive cord. No terminal filament is present. Late‐stage adult females show synchronized development of the ovarioles, while in undernourished females, a small proportion of ovarioles proceed to maturity.

Dimorphic ovary differentiation in honeybee (Apis mellifera) larvae involves caste-specific expression of homologs of ark and buffy cell death genes.

The establishment of the number of repeated structural units, the ovarioles, in the ovaries is one of the critical events that shape caste polyphenism in social insects. In early postembryonic development, honeybee (Apis mellifera) larvae have a pair of ovaries, each one consisting of almost two hundred ovariole primordia. While practically all these ovarioles continue developing in queen-destined larvae, they undergo massive programmed cell death (PCD) in worker-destined larvae. So as to gain insight into the molecular basis of this fundamental process in caste differentiation we used quantitative PCR (qPCR) and fluorescent in situ hybridization (FISH) to investigate the expression of the Amark and Ambuffy genes in the ovaries of the two honeybee castes throughout the fifth larval instar. These are the homologs of ark and buffy Drosophila melanogaster genes, respectively, involved in activating and inhibiting PCD. Caste-specific expression patterns were found during this time-window defining ovariole number. Amark transcript levels were increased when ovariole resorption was intensified in workers, but remained at low levels in queen ovaries. The transcripts were mainly localized at the apical end of all the worker ovarioles, but appeared in only a few queen ovarioles, thus strongly suggesting a function in mediating massive ovariolar cell death in worker larvae. Ambuffy was mainly expressed in the peritoneal sheath cells covering each ovariole. The levels of Ambuffy transcripts increased earlier in the developing ovaries of queens than in workers. Consistent with a protective role against cell death, Ambuffy transcripts were localized in practically all queen ovarioles, but only in few worker ovarioles. The results are indicative of a functional relationship between the expression of evolutionary conserved cell death genes and the morphological events leading to caste-specific ovary differentiation in a social insect.

Cheating workers with large activated ovaries avoid risky foraging

Eusocial insects undoubtedly evolved from solitary ancestors, but how this occurred is not well established. The Ground Plan hypothesis suggests that gene networks that once regulated the oviposition and foraging phases of an ancestral solitary insect’s life cycle have been co-opted to establish the queen–worker dimorphism in extant eusocial insects; queens permanently express genes that were once expressed during the oviposition phase, whereas workers express genes that were once associated with foraging. An extension of the Ground Plan hypothesis, the Reproductive Ground Plan-forager hypothesis, proposes that foraging specialization by worker honey bees for either pollen or nectar is controlled by the same reproductive gene networks. According to the Reproductive Ground Plan-forager hypothesis, workers with more ovarioles forage early in life and specialize in pollen collection. Here we find that among workers of a highly reproductive honey bee subspecies, Apis mellifera capensis, there is a positive correlation between ovariole number and age at onset of foraging, and no association between ovariole number and foraging preference, thus contradicting key aspects of the Reproductive Ground Plan-forager hypothesis. We also find a negative association between ovariole number and ovary activation, suggesting that high ovariole number is not directly related to reproductive potential as previously assumed.

Insulin signalling underlies both plasticity and divergence of a reproductive trait in Drosophila.

Phenotypic plasticity is the ability of a single genotype to yield distinct phenotypes in different environments. The molecular mechanisms linking phenotypic plasticity to the evolution of heritable diversification, however, are largely unknown. Here, we show that insulin/insulin-like growth factor signalling (IIS) underlies both phenotypic plasticity and evolutionary diversification of ovariole number, a quantitative reproductive trait, in Drosophila. IIS activity levels and sensitivity have diverged between species, leading to both species-specific ovariole number and species-specific nutritional plasticity in ovariole number. Plastic range of ovariole number correlates with ecological niche, suggesting that the degree of nutritional plasticity may be an adaptive trait. This demonstrates that a plastic response conserved across animals can underlie the evolution of morphological diversity, underscoring the potential pervasiveness of plasticity as an evolutionary mechanism.

Nurse bee behaviour manipulates worker honeybee (Apis mellifera L.) reproductive development

The evolution of nonreproductive castes is a fundamental question in evolution biology. The honeybee Apis mellifera L. has a reproductive division of labour: the queen is the primary egg-layer in a colony and has more than 200 ovarian filaments (ovarioles), whereas a worker normally does not reproduce and has fewer than 20 ovarioles. The number of ovarioles influences worker foraging behaviour and the propensity to become an egg-layer in the absence of the queen, suggesting that reproductive regulatory networks evolved with foraging division of labour in honeybee workers. Cooperation between nurse bee feeding behaviour and larval developmental programming results in the differentiation of queens and workers along with variation in ovariole number, body mass and foraging behaviour. Here, we tested how nurse bees affect ovariole number and body mass in workers, and how larvae respond to food delivery during different larval life stages. Our findings demonstrate that nurses control larvae growth and ovariole number by temporally manipulating food delivery and that the response of larvae to food differs with larval life stage and genotype. Body mass of larvae was more sensitive to nutrition during the first to the fourth instar (L1–L4), whereas ovariole number was more sensitive during the fifth instar (L5). Overall, we were able to decouple the nurse feeding program and the larval development program in honeybees. We conclude that nurse feeding behaviour during L5 is critical for modulating ovariole number in workers.

SIMILARITIES AND DIFFERENCES IN ALTITUDINAL VERSUS LATITUDINAL VARIATION FOR MORPHOLOGICAL TRAITS INDROSOPHILA MELANOGASTER

Understanding how natural environments shape phenotypic variation is a major aim in evolutionary biology. Here, we have examined clinal, likely genetically based variation in morphology among 19 populations of the fruit fly (Drosophila melanogaster) from Africa and Europe, spanning a range from sea level to 3000 m altitude and including locations approximating the southern and northern range limit. We were interested in testing whether latitude and altitude have similar phenotypic effects, as has often been postulated. Both latitude and altitude were positively correlated with wing area, ovariole number, and cell number. In contrast, latitude and altitude had opposite effects on the ratio between ovariole number and body size, which was negatively correlated with egg production rate per ovariole. We also used transgenic manipulation to examine how increased cell number affects morphology and found that larger transgenic flies, due to a higher number of cells, had more ovarioles, larger wings, and, unlike flies from natural populations, increased wing loading. Clinal patterns in morphology are thus not a simple function of changes in body size; instead, each trait might be subject to different selection pressures. Together, our results provide compelling evidence for profound similarities as well as differences between phenotypic effects of latitude and altitude.