RESUMEN
The health of the honey bee Apis mellifera is challenged by the ectoparasitic mite Varroa destructor, and the numerous harmful pathogens it vectors. Existing pesticide-based Varroa controls are not sustainable. In contrast, one promising approach for improved honey bee health is the breeding of hygienic bees, capable of detecting and removing brood that is parasitized or diseased. In three experiments we find evidence to support the hypothesis that stock-specific chemical brood signals are induced by Varroa and Deformed Wing Virus, and elicit hygienic response in the honey bee. By collecting, analyzing, and running bioassays involving mite-infested and control brood extracts from three honey bee breeding stocks we: 1) found evidence that a transferrable chemical signal for hygienic behavior is present in Varroa-infested brood extracts, 2) identified ten stock-specific hydrocarbons as candidates of hygienic signaling, and 3) found that two of these hydrocarbons linked to Varroa and DWV were also elevated in brood targeted for hygienic behavior. These findings expand our understanding of honey bee chemical communication, and facilitate the development of improved hygienic selection tools to breed honey bees with greater resistance to Varroa and associated pathogens.
Asunto(s)
Comunicación Animal , Abejas , Aseo Animal , Hidrocarburos/metabolismo , Virus ARN , Varroidae , Animales , Abejas/parasitología , Abejas/virologíaRESUMEN
Although extensive research has been carried out on the desert ants in the genus Cataglyphis in recent years, some of the specific intra- and interspecific relationships remain elusive. The present study disentangles the phylogenetic relationships among the C. bicolor complex in Israel using an integrative approach based on genetic markers, morphometric measurements, and chemical analyses (cuticular hydrocarbons). Several species delimitation approaches based on four nuclear, two mitochondrial, and eleven microsatellite markers, as well as 16 body measurements and 56 chemical variables, were employed to deciphering the occurrence of cryptic species in our data set. Our findings support the occurrence of at least four distinct species in the C. bicolor group in Israel, one of which may be a complex of three more recent species. The findings confirm the distinctiveness of C. isis and C. holgerseni. They attest the presence of a recently discovered species, C. israelensis, in the central mountain ridge and the occurrence of another clade distributed from the Negev to the Mediterranean coast, comprising the species C. niger, C. savignyi, and C. drusus. Although these three species are separated on the basis of mtDNA, this subgrouping was not supported by any of the nuclear sequence markers nor by the microsatellite analysis. This genetic structure may thus either reflect a possible recent speciation, or a geographical structuring of a single species. Overall, using these different sources of evidence we locate our samples within a global phylogeny of the bicolor group and discuss the processes that underlie speciation in this group.
Asunto(s)
Hormigas/clasificación , Animales , Hormigas/química , Hormigas/genética , Citocromos b/clasificación , Citocromos b/genética , ADN Mitocondrial/genética , ADN Mitocondrial/metabolismo , Complejo IV de Transporte de Electrones/clasificación , Complejo IV de Transporte de Electrones/genética , Haplotipos , Israel , Repeticiones de Microsatélite/genética , Filogenia , Análisis de Componente Principal , Especificidad de la EspecieRESUMEN
Juvenile hormone (JH) is an important regulator of development and physiology in insects. While in many insect species, including bumble bees, JH functions as gonadotropin in adults, in some highly eusocial insects its role has shifted to regulate social behavior including division of labor, dominance and aggression. Studying JH functions across social insect species is important for understanding the evolution of sociality; however, these studies have been limited because of the inability to reduce JH levels without surgically removing its glandular source, the corpora allata. Precocene is known to inhibit JH biosynthesis in several non-social insects, but has been poorly studied in social insects. Here, we tested whether precocene-I can effectively reduce JH levels in Bombus terrestris workers, and examined its effects on their physiology and behavior. Precocene-I treatment of three-worker groups decreased JH titer and ovarian activation, irrespective of the bees' dominance rank within the group, and was remedied by JH replacement therapy. Precocene-I also decreased aggressiveness and increased ester-sterility signal production; these changes were rank-dependent, and affected mainly the most reproductive and the least aggressive workers, respectively, and could not be remedied by JH replacement therapy. These results clearly confirm the role of JH as a gonadotropin and mediator of aggression in B. terrestris, and indicate that JH effects are associated with worker dominance rank. The ability to chemically reduce JH titer provides us with a non-intrusive method to probe the evolutionary changes associated with JH and the hormonal mechanisms that are associated with reproduction and behavior in social insects.
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Agresión/efectos de los fármacos , Abejas/efectos de los fármacos , Abejas/fisiología , Benzopiranos/farmacología , Hormonas Juveniles/biosíntesis , Hormonas Juveniles/fisiología , Ovario/efectos de los fármacos , Ovario/fisiología , Conducta Social , Animales , Conducta Animal/efectos de los fármacos , Conducta Animal/fisiología , Femenino , Infertilidad Femenina , Reproducción/efectos de los fármacosRESUMEN
In their invasive ranges, Argentine ant populations often form one geographically vast supercolony, genetically and chemically uniform within which there is no intraspecific aggression. Here we present regional patterns of intraspecific aggression, cuticular hydrocarbons (CHCs) and population genetics of 18 nesting sites across Corsica and the French mainland. Aggression tests confirm the presence of a third European supercolony, the Corsican supercolony, which exhibits moderate to high levels of aggression, depending on nesting sites, with the Main supercolony, and invariably high levels of aggression with the Catalonian supercolony. The chemical analyses corroborated the behavioural data, with workers of the Corsican supercolony showing moderate differences in CHCs compared to workers of the European Main supercolony and strong differences compared to workers of the Catalonian supercolony. Interestingly, there were also clear genetic differences between workers of the Catalonian supercolony and the two other supercolonies at both nuclear and mitochondrial markers, but only very weak genetic differentiation between nesting sites of the Corsican and Main supercolonies (F(ST) = 0.06). A detailed comparison of the genetic composition of supercolonies also revealed that, if one of the last two supercolonies derived from the other, it is the Main supercolony that derived from the Corsican supercolony rather than the reverse. Overall, these findings highlight the importance of conducting more qualitative and quantitative analyses of the level of aggression between supercolonies, which has to be correlated with genetic and chemical data.
Asunto(s)
Agresión , Hormigas/fisiología , Variación Genética , Hidrocarburos/análisis , Especies Introducidas , Animales , Hormigas/genética , Europa (Continente) , Francia , Genética de Población , Repeticiones de Microsatélite , Datos de Secuencia MolecularRESUMEN
In social Hymenoptera, within-colony relatedness is usually high due to the haplodiploid sex-determining system. However, factors such as the presence of multiple reproductive queens (polygyny), multiple queen matings (polyandry) or worker reproduction result in decreased relatedness among workers and the brood they rear, and consequently dilute their inclusive fitness benefits from helping. Here, we investigated population genetic structure, mating system, worker reproduction and parthenogenesis in the desert ant Cataglyphis sabulosa. Analysis of worker genotypes showed that colonies are headed by a single queen, mated with 1-5 males. The inbreeding coefficient within colonies and the levels of relatedness between the queens and their mates were positive, indicating that mating occurs between related individuals. Moreover, the mates of a queen are on average related and contribute equally to worker production. Our analyses also indicate that colonies are genetically differentiated and form a population exhibiting no isolation-by-distance pattern, consistent with the independent foundation of new colonies (that is, without the help of workers). Finally, both ovarian dissections and genetic data on the parentage of males show that workers do not reproduce in queenright colonies; however, they lay both haploid (arrhenotokous males) and diploid (thelytokous females) eggs in queenless colonies. In contrast to the congeneric species C. cursor, where new queens are produced by thelytokous parthenogenesis, female sexuals of C. sabulosa result from classical sexual reproduction.
Asunto(s)
Hormigas/genética , Animales , Hormigas/fisiología , Femenino , Masculino , PartenogénesisRESUMEN
In the honeybee, Apis mellifera, the queen monopolizes reproduction, while the sterile workers cooperate harmoniously in nest maintenance. However, under queenless (QL) conditions, cooperation collapses and reproductive competition among workers ensues. This is mediated through aggression and worker oviposition, as well as shifts in pheromones, from worker to queen-like composition. Many studies suggest a dichotomy between conflict resolution through aggression or through pheromonal signaling. In this paper, we demonstrate that both phenomena comprise essential components of reproductive competition and that pheromone signaling actually triggers the onset of aggression. We kept workers as QL groups until first aggression was observed and subsequently determined the contestants' reproductive status and content of the mandibular (MG) and Dufour's glands (DG). In groups in which aggression occurred early, the attacked bee had consistently more queen-like pheromone in both the MG and DG, although both contestants had undeveloped ovaries. In groups with late aggression, the attacked bee had consistently larger oocytes and more queen-like pheromone in the DG, but not the MG. We suggest that at early stages of competition, the MG secretion is utilized to establish dominance and that the DG provides an honest fertility signal. We further argue that it is the higher amount of DG pheromone that triggers aggression.
Asunto(s)
Agresión , Abejas/fisiología , Conducta Animal , Feromonas/fisiología , Transducción de Señal/fisiología , Predominio Social , Animales , Femenino , Feromonas/metabolismo , Reproducción , Glándulas Sebáceas/metabolismoRESUMEN
Chemical analyses revealed that the labial gland complex of worker honeybees possesses a series of hydrocarbons dominated by odd-numbered carbon chain alkanes along with minor amounts of alkenes and branched alkanes. Foragers contained significantly more secretion than nurse bees. Experiments with bees from colonies induced to have a division of labor independent of age revealed that the differences in the amount of secretion were task, but not age dependent.
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Abejas/fisiología , Conducta Animal , Hidrocarburos/química , Glándulas Salivales/química , Factores de Edad , Animales , Masculino , VolatilizaciónRESUMEN
There are two separate, and presumably opposing, processes that affect colony odor in the desert ant Cataglyphis niger: (1) biosynthesis and turnover of these chemicals by individual ants, and (2) homogenization of colony odor through exchange of cues. The first increases signal variability; the latter decreases it. The impact of these factors was tested by splitting colonies and monitoring the profile changes occurring in the postpharyngeal glands (PPG) and cuticular hydrocarbons. From each of two polygynous nests four daughter colonies were formed, three monogynous and one queenless. Thereafter, 10 ants from each were randomly selected each month, for three successive months, for analyses of their PPG and cuticular hydrocarbons. From two colonies we also obtained ants from a known matriline. Over time, there was a shift in hydrocarbon profiles of both the PPG and cuticular washes in each of the tested colonies. Moreover, by subjecting selected hydrocarbon constituents to a discriminant analyses based on their relative proportions, all of the daughter colonies (queenright and queenless) were distinguishable from each other and from their respective mother colonies. In each of the queenright daughter colonies, the queen profile was indiscriminable from that of the workers and often was in the center of the group. Full sisters were clearly distinguishable from their nestmates, emphasizing the genetic versus environmental processes that govern colony odor. The effect of time was always superior to the separation effect in contributing to odor segregation. Comparison of the Mahalanobis distances indicated that the shift in hydrocarbon seems to proceed along parallel lines rather than in divergence. However, there was no overt aggression between ants that originated from the different subgroups in dyadic encounters. It appears that in this species a three-month separation pe riod is not sufficient to change the hydrocarbon profile beyond the recognition threshold.
Asunto(s)
Hormigas/fisiología , Hidrocarburos/química , Odorantes/análisis , Animales , Conducta Animal , Evolución Biológica , Ambiente , Femenino , MasculinoRESUMEN
In the social bee, Bombus terrestris, the two castes differ in size and physiology, but not in any other morphological and anatomical aspects. The size differences between the castes are the result of longer instar duration in prospective queen larvae. It appears that queen larvae are programmed to have a higher molting weight at the end of the 2nd, 3rd and 4th instars. Calculation of the growth ratio, the ratio between the logarithm of molting weight at two successive instars, revealed that queen larvae have a linear growth ratio over the entire larval development as predicted by Dyar's rule. In the worker larvae, in contrast, linearity of the growth ratio breaks after the second instar, resulting in larval molting at lower weights than expected by Dyar's rule. We therefore suggest that workers' development is abnormally shortened, either by parental manipulation or by adopting a different growth plan in response to the queen's signal.
Asunto(s)
Abejas , Conducta Social , Animales , Abejas/anatomía & histología , Abejas/fisiología , Peso Corporal , Femenino , Larva , Especificidad de la EspecieRESUMEN
The chemical strategies by which parasites manage to break into the social fortresses of ants offer a fascinating theme in chemical ecology. Semiochemicals used for interindividual nestmate recognition are also involved in the mechanisms of tolerance and association between the species, and social parasites exploit these mechanisms. The obligate parasites are odorless ("chemical insignificance") at the time of usurpation, like all other callow ants, and this "invisibility" enables their entry into the host colony. By chemical mimicry (sensu lato), they later integrate the gestalt odor of this colony ("chemical integration"). We hypothesize that host and parasite are likely to be related chemically, thereby facilitating the necessary mimicry to permit bypassing the colony odor barrier. We also review the plethora of chemical weapons used by social parasites (propaganda, appeasement, and/or repellent substances), particularly during the usurpation period, when the young mated parasite queen synthesizes these chemicals before usurpation and ceases such biosynthesis afterwards. We discuss evolutionary trends that may have led to social parasitism, focusing on the question of whether slave-making ants and their host species are expected to engage in a coevolutionary arms race.
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Hormigas/química , Hormigas/parasitología , Conducta Social , Animales , Ecología , Interacciones Huésped-Parásitos , Modelos BiológicosRESUMEN
Mounting evidence implicates ecdysteroids in queen-worker differentiation during the last larval instars of highly social insects. In the present study, we analyzed ecdysteroid titers in queen and worker larvae of the bumblebee Bombus terrestris from the second to the early fourth instar. B. terrestris is of particular interest because caste is already determined in the second instar, presumably by a pheromonal signal emitted by the egg-laying queen. Caste differences in the adults, however, are only expressed at the physiological and not at the morphological level, except for the distinctly larger size of the queen. In the second and third instar, ecdysteroid titers in queen larvae were generally higher than those of workers. These early caste-specific differences, however, were abolished in the fourth instar. In the early fourth instar we could detect two small ecdysteroid peaks, with the one preceding the cocoon-spinning phase presenting the characteristics of a pupal commitment peak. The synchrony of caste differences in ecdysteroid and juvenile hormone titers suggests a synergistic action of these hormones in caste determination.
RESUMEN
Our aim was to test the existence of Gestalt colony odour in Camponotus fellah. We isolated individual workers to prevent trophallaxis, allogrooming and body contact. After 20 days, the cuticular hydrocarbon profile of the isolated ants diverged from that of the parent colony. Moreover, each isolated individual had its own specific blend. This procedure showed that after about 20 days of isolation there was a turnover of the colony odour, revealing the genetically expressed hydrocarbon profile of each individual. It also showed that the cuticular hydrocarbon profile is polymorphic, and that its homogeneity within a colony is maintained by frequent exchanges of hydrocarbons between workers. Behavioural observations of resident workers, in their nest, towards nestmates reintroduced after isolation indicated that a short isolation period (3-5 days), which induced a minor change in hydrocarbon profile, provoked frequent trophallactic solicitations. These were likely to permit the isolated ants to readjust their hydrocarbon profile to that of the ants in the mother colony. Longer isolation periods (20-40 days) induced a greater change in hydrocarbon profile and made the residents intolerant towards their introduced nestmates. Therefore, our results clearly support the existence of a Gestalt colony odour in C. fellah. They also show that since individual hydrocarbon production is dynamic, workers are obliged to exchange hydrocarbons continually (mainly by trophallaxis) in order to be in the Gestalt, and properly integrate into the colony. Copyright 2000 The Association for the Study of Animal Behaviour.
RESUMEN
Chemical analyses revealed that in Cataglyphis niger both the hemolymph and the crop contain the same hydrocarbons that are found in the postpharyngeal gland (PPG). On the cuticle, on the other hand, alkanes, and in particular nonacosane, were more abundant than in the PPG. Studies of their biosynthesis in vivo, using intact ants, revealed the presence of newly synthesized hydrocarbons in both the PPG and the crop. In decapitated ants (in the absence of the PPG), however, the crop did not contain any newly synthesized hydrocarbons, indicating the PPG as the major source of crop hydrocarbons. The fat body, as demonstrated by in vitro studies, is the major tissue that biosynthesizes hydrocarbons. The PPG failed to do so, but showed good de novo biosynthesis of other lipid constituents. The large amount of hydrocarbons in the crop suggests that the alimentary canal may serve as an outlet for the overflow of PPG hydrocarbons, or as a route for the directed clearance of hydrocarbons from the PPG.These results confirm and enlarge the model proposed for hydrocarbon circulation in C. niger. They are synthesized by the fat body, released to the hemolymph and transported to the cuticle and the PPG. The PPG hydrocarbons are applied to the cuticle by self-grooming, but can also be cleared via the alimentary canal. Partial emptying of the PPG may facilitate the admixing of recognition cues that the ant may acquire from nestmates by trophallaxis. The reason for the dissimilarity in hydrocarbon composition between the PPG and the cuticle is not yet clear; it may be due to secretions from additional glands, or reflect deviant hydrocarbon transport mechanisms between the PPG and the cuticle.
RESUMEN
Plasticity of Dufour's gland secretion in the honey bee is correlated with the individual's plasticity. Queens and queenless (QL) egg-laying workers possess a bouquet of esters and hydrocarbons, whereas queenright (QR) workers produce exclusively hydrocarbons. The effects of social environment (QR vs. QL conditions) and possible physiological constraints on the gland were studied by following the biosynthesis of these classes of compounds in vivo and in vitro. Biosynthesis in vivo followed the prediction based on glandular chemistry. Queens and QL egg-laying workers, but not QR workers or QL foragers, showed incorporation of sodium acetate into both hydrocarbons and esters. In contrast, the in vitro studies revealed that, in addition to queens and QL egg-laying workers, QR nurses retained their ability to produce the queen characteristic esters. Although there was some ester production in foragers, it occurred to a lesser extent. It is possible that the glands in the older foragers undergo irreversible changes. The in vitro incubation also revealed a temporal activation of ester biosynthesis in QR workers. In these glands alcohols, corresponding to the alcohol moiety of the esters, predominated in short-term incubations but decreased as the amount of newly synthesized esters increased. In contrast, queens and QL egg-laying workers showed predominant incorporation into esters from the onset of incubation. Thus, expression within the workers' Dufour's gland is regulated. In the presence of a queen, ester production is inhibited. Once the queen is removed the physiologically unconstrained gland starts to biosynthesize the queen-specific esters after a certain lag needed for the build-up of precursors and the enzymatic machinery.
RESUMEN
The role of ecdysteroids in the regulation of dominance and reproduction in social Hymenoptera is little explored. In the current study we compared ecdysteroid titers in hemolymph of individual queen and worker bumble bees (Bombus terrestris) that differ in their behavior, reproductive status and social environment. Egg-laying queens that head colonies and have ovaries exhibiting all stages of follicle development, had a higher ecdysteroid titer than virgin queens whose ovaries contained only follicles at initial stages. In workers, the relationship between ecdysteroid titers and follicle development appears to be more complex and to be influenced by the bee's social status and social environment. Shortly after emergence, young workers had only follicles at the initial stages of oogenesis and they exhibited a low ecdysteroid titer. No significant correlation was detected between ovary status and ecdysteroid titer in workers, with some workers showing activated ovaries but low ecdysteroid titers. However, at six days of age, a trend towards higher ecdysteroid titer was observed for workers in queenless groups, a condition characterized by rapid follicle development relative to queenright conditions. In these queenless groups, high social status was associated with high ecdysteroid titers. By contrast, in queenright workers ecdysteroid titers were low, even for bees with presumably high social status that had activated ovaries and were observed performing oviposition behavior. This study suggests that ecdysteroids are involved in regulation of reproduction in B. terrestris.
RESUMEN
To begin to explore the role of biogenic amines in reproductive division of labor in social insects, brain levels of dopamine, serotonin, and octopamine were measured in bumble bee (Bombus terrestris) workers and queens that differ in behavioral and reproductive state. Levels of all three amines were similar for mated and virgin queens. Young workers that developed with or without a queen had similar amine levels, but in queenright colonies differences in biogenic amine levels were associated with differences in behavior and reproductive physiology. Dominant workers had significantly higher octopamine levels compared with workers of lower dominance status but of similar size, age, and ovary state. High dopamine levels were associated with the last stages of oocyte development irrespective of worker social status and behavior. These results suggest that biogenic amines are involved in behavioral and physiological aspects of regulation of reproduction in bumble bees.
Asunto(s)
Abejas , Aminas Biogénicas/farmacología , Conducta Sexual Animal , Predominio Social , Animales , Química Encefálica , Femenino , Masculino , ReproducciónRESUMEN
Social deprivation is an unusual situation for ants that normally maintain continuous contact with their nestmates. When a worker was experimentally isolated for 5 days and then reunited with a nestmate, she engaged in prolonged trophallaxis. It is suggested that trophallaxis allows her to restore a social bond with her nestmates and to re-integrate into the colony, particularly via the exchange of colony-specific hydrocarbons. Octopamine reduced trophallaxis in these workers as well as hydrocarbon transfer between nestmates, but not hydrocarbon biosynthesis. Administration of serotonin to such 5-day-isolated ants had no effect on the percentage of trophallaxis. Administration of phentolamine alone, an octopamine antagonist, had no effect, but when co-administrated with octopamine it reduced the effect of octopamine alone and restored trophallaxis to control levels. Moreover, the observed effect of octopamine was not due to a non-specific effect on locomotor activity. Therefore, we hypothesise that octopamine mediates behaviour patterns linked to social bonding, such as trophallaxis. On the basis of an analogy with the role of norepinephrine in vertebrates, we suggest that the levels of octopamine in the brain of socially deprived ants may decrease, together with a concomitant increase in their urge to perform trophallaxis and to experience social contacts. Octopamine administration may reduce this social deprivation effect, and octopamine could therefore be regarded as being partly responsible for the social cohesion between nestmates in ant colonies.
Asunto(s)
Hormigas/fisiología , Comunicación Animal , Animales , Hormigas/metabolismo , Hidrocarburos/metabolismo , Actividad Motora/efectos de los fármacos , Octopamina/antagonistas & inhibidores , Octopamina/farmacología , Octopamina/fisiología , Fentolamina/farmacología , Serotonina/farmacología , Aislamiento SocialRESUMEN
The effects of the social environment and age on juvenile hormone (JH) and reproduction were investigated by measuring ovarian development, hemolymph levels of JH III, and rates of JH biosynthesis from the same individual bumble bees (Bombus terrestris). Differences in social environment were associated with differences in rates of JH biosynthesis, JH titer and ovarian development. Young queenless workers had a higher rate of JH biosynthesis, JH titer and ovarian development than queenright (QR) workers of similar age. Dominant workers in QR colonies had a higher rate of JH biosynthesis, JH titer and ovarian development than low ranked workers of similar size. There was a positive correlation between JH titer and ovarian development, but no correlation between rate of JH biosynthesis and ovarian development or between JH biosynthesis and JH titer. Both JH titer and rate of JH biosynthesis increased with age from emergence to 3 days of age, but 6-day-old workers, egg-laying workers, and actively reproducing queens had high JH titers and highly developed ovaries but low rates of JH biosynthesis. These results show that reproduction in B. terrestris is strongly affected by the social environment and the influence of the environment on reproduction is mediated by JH. Our data also indicate that the rate of JH biosynthesis measured in vitro is not a reliable indicator of JH titer or ovarian development in B. terrestris; possible reasons are discussed.
RESUMEN
The critical period for caste determination and its juvenile hormone (JH III) correlates were studied in Bombus terrestris. Larvae of known age and instar were taken from young colonies, in which they would have been reared as workers, and placed into groups of queenless workers. Under these conditions the critical age for caste determination was 5 days, during the second instar. Endocrine correlates of caste determination were obtained by determining profiles of juvenile hormone titer and juvenile hormone biosynthesis, measured by chiral-specific radioimmunoassay and the in vitro radiochemical assay, respectively. By the middle of the second instar prospective queen larvae had significantly higher rates of juvenile hormone biosynthesis and juvenile hormone titer than prospective worker larvae. Based on the coincidence of timing of both the critical period and the appearance of caste-specific juvenile hormone titer, we suggest that juvenile hormone plays a role in the mechanisms that control caste determination in B. terrestris.
Asunto(s)
Abejas/fisiología , Jerarquia Social , Hormonas Juveniles/fisiología , Animales , Abejas/anatomía & histología , Abejas/crecimiento & desarrollo , Peso Corporal , Hormonas Juveniles/biosíntesis , Larva/anatomía & histología , Larva/fisiología , Muda , Factores de TiempoRESUMEN
To study the possible role of juvenile hormone in caste determination in Bombus terrestris, we measured development and rates of juvenile hormone biosynthesis in vitro in larvae destined to develop into either workers or queens. Larvae of both castes developed through four instars and had the same growth rates. However, the duration of the instars was longer for queen larvae, and their head width at the third and fourth instars was significantly larger. After validating the well-known radiochemical assay of JH for bumble bee larvae, we show that worker larvae corpora allata exhibited a constant and low rate of JH biosynthesis, never more than 5 pmol JH/h/pair. Queen larvae, in contrast, had two peaks of JH biosynthesis: a small one during the first instar, which has previously been correlated with caste determination; and a large peak, previously undetected, above 40 pmol JH/h/pair, during the second and third instars. We suggest that caste determination in this species is mediated by JH and that the duration of larval instars is a key factor. The possibility that the queen influences caste determination via an effect on instar duration is also discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved