Effects of Insemination Quantity on Honey Bee Queen Physiology

Mating has profound effects on the physiology and behavior of female insects, and in honey bee (Apis mellifera) queens, these changes are permanent. Queens mate with multiple males during a brief period in their early adult lives, and shortly thereafter they initiate egg-laying. Furthermore, the pheromone profiles of mated queens differ from those of virgins, and these pheromones regulate many different aspects of worker behavior and colony organization. While it is clear that mating causes dramatic changes in queens, it is unclear if mating number has more subtle effects on queen physiology or queen-worker interactions; indeed, the effect of multiple matings on female insect physiology has not been broadly addressed. Because it is not possible to control the natural mating behavior of queens, we used instrumental insemination and compared queens inseminated with semen from either a single drone (single-drone inseminated, or SDI) or 10 drones (multi-drone inseminated, or MDI). We used observation hives to monitor attraction of workers to SDI or MDI queens in colonies, and cage studies to monitor the attraction of workers to virgin, SDI, and MDI queen mandibular gland extracts (the main source of queen pheromone). The chemical profiles of the mandibular glands of virgin, SDI, and MDI queens were characterized using GC-MS. Finally, we measured brain expression levels in SDI and MDI queens of a gene associated with phototaxis in worker honey bees (Amfor). Here, we demonstrate for the first time that insemination quantity significantly affects mandibular gland chemical profiles, queen-worker interactions, and brain gene expression. Further research will be necessary to elucidate the mechanistic bases for these effects: insemination volume, sperm and seminal protein quantity, and genetic diversity of the sperm may all be important factors contributing to this profound change in honey bee queen physiology, queen behavior, and social interactions in the colony.

[1]  M. Winston,et al.  EFFECTS OF HONEY BEE ( APIS MELLIFERA L.) QUEEN MANDIBULAR GLAND PHEROMONE ON FORAGING AND BROOD REARING , 1992 .

[2]  M. Winston,et al.  Semiochemical basis of the retinue response to queen honey bees , 1988, Nature.

[3]  A. Collins,et al.  Sperm storage and antioxidative enzyme expression in the honey bee, Apis mellifera , 2004, Insect molecular biology.

[4]  J. M. Graham,et al.  The hive and the honey bee. , 1992 .

[5]  B. Oldroyd,et al.  Effects of Intracolonial Genetic Diversity on Honey Bee (Hymenoptera: Apidae) Colony Performance , 1992 .

[6]  Mark L. Winston,et al.  The effect of queen pheromones on worker honey bee ovary development , 2003, Naturwissenschaften.

[7]  L. Roth.  Hypersexual Activity Induced in Females of the Cockroach Nauphoeta cinerea , 1962, Science.

[8]  M. Wolfner.  The gifts that keep on giving: physiological functions and evolutionary dynamics of male seminal proteins in Drosophila , 2002, Heredity.

[9]  C. Gillott.  Male accessory gland secretions: modulators of female reproductive physiology and behavior. , 2003, Annual review of entomology.

[10]  T. Seeley,et al.  Lower disease infections in honeybee (Apis mellifera) colonies headed by polyandrous vs monandrous queens , 2006, Naturwissenschaften.

[11]  M. Winston,et al.  Selection on worker honeybee responses to queen pheromone (Apis mellifera L.) , 2000, Naturwissenschaften.

[12]  Ken Sasaki,et al.  Depression of brain dopamine and its metabolite after mating in European honeybee (Apis mellifera) queens , 2005, Naturwissenschaften.

[13]  M. Winston,et al.  Suppression of queen rearing in European and Africanized honey beesApis mellifera L. by synthetic queen mandibular gland pheromone , 1995, Insectes Sociaux.

[14]  P. Neumann,et al.  Multiple nuptial flights, sperm transfer and the evolution of extreme polyandry in honeybee queens , 2005, Animal Behaviour.

[15]  R. Crozier,et al.  Polyandry in the genus Apis, particularly Apis andreniformis , 1997, Behavioral Ecology and Sociobiology.

[16]  J. Woyke.  Dynamics of Entry of Spermatozoa into the Spermatheca of Instrumentally inseminated Queen Honeybees , 1983 .

[17]  E. Morgan,et al.  Chemistry of a Mating Plug in Bumblebees , 2000, Journal of Chemical Ecology.

[18]  H. H. Laidlaw,et al.  Queen Rearing and Bee Breeding , 1997 .

[19]  N. Koeniger,et al.  Proteins in spermathecal gland secretion and spermathecal fluid and the properties of a 29 kDa protein in queens of Apis mellifera , 2004 .

[20]  Mark L. Winston,et al.  Pheromone Communication in the Honeybee (Apis mellifera L.) , 2005, Journal of Chemical Ecology.

[21]  H. Nijhout.  Abdominal stretch reception in Dipetalogaster maximus (Hemiptera: Reduviidae) , 1984 .

[22]  M. Winston,et al.  Species- and Caste-Determined Mandibular Gland Signals in Honeybees (Apis) , 1997, Journal of Chemical Ecology.

[23]  P. Schmid-Hempel,et al.  A nonspecific fatty acid within the bumblebee mating plug prevents females from remating , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Y. Ben-Shahar.  The foraging gene, behavioral plasticity, and honeybee division of labor , 2005, Journal of Comparative Physiology A.

[25]  O. Kaftanoğlu,et al.  Effects of insemination on the initiation of oviposition in the queen honeybee. , 1982 .

[26]  A. Mercer,et al.  The effects of queenlessness on the maturation of the honey bee olfactory system , 1998, Behavioural Brain Research.

[27]  J. Lobo,et al.  Estimation of the number of matings in Apis mellifera: extensions of the model and comparison of different estimates , 1993 .

[28]  J. Ringo.  Sexual receptivity in insects. , 1996, Annual review of entomology.

[29]  K. Hartfelder,et al.  The initial stages of oogenesis and their relation to differential fertility in the honey bee (Apis mellifera) castes. , 2004, Arthropod structure & development.

[30]  M. Winston,et al.  Variation in worker response to honey bee (Apis mellifera L.) queen mandibular pheromone (Hymenoptera: Apidae) , 2005, Journal of Insect Behavior.

[31]  J. Woyke,et al.  FURTHER INVESTIGATIONS ON NATURAL MATING OF INSTRUMENTALLY INSEMINATED APIS MELLIFERA QUEENS , 1995 .

[32]  H. A. Dade.  Anatomy and Dissection of the Honeybee , 1962 .

[33]  R. Crozier,et al.  Evolution of Mating Behavior in the Genus Apis and an Estimate of Mating Frequency in Apis cerana (Hymenoptera: Apidae) , 1998 .

[34]  M. Winston,et al.  Queen attendance behavior of worker honey bees (Apis mellifera L.) that are high and low responding to queen mandibular pheromone , 1995, Insectes Sociaux.

[35]  T. Seeley,et al.  Honeybee Ecology: A Study of Adaptation in Social Life , 1985 .

[36]  M. Winston,et al.  Semiochemicals of the honeybee queen mandibular glands , 1990, Journal of Chemical Ecology.

[37]  Gene E Robinson,et al.  Pheromone-mediated gene expression in the honey bee brain , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Winston,et al.  New components of the honey bee (Apis mellifera L.) queen retinue pheromone , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[39]  Christina M. Grozinger,et al.  Uncoupling primer and releaser responses to pheromone in honey bees , 2007, Naturwissenschaften.

[40]  R. Crozier,et al.  Evolution of social insect colonies , 1996 .

[41]  M. Blum,et al.  Chemistry of the Drone Honey Bee Reproductive System. II. Carbohydrates in the Reproductive Organs and Semen , 1962 .

[42]  M. Winston,et al.  Anarchistic queen honey bees have normal queen mandibular pheromones , 2005, Insectes Sociaux.

[43]  J. Free,et al.  Pheromones of social bees , 1987 .

[44]  P. Kulinowski,et al.  Magnetic resonance microscopy of internal structure of drone and queen honey bees , 1996 .

[45]  L. Keller,et al.  The role of queen pheromones in social insects: queen control or queen signal? , 1993, Animal Behaviour.

[46]  M. Winston,et al.  EFFECT OF QUEEN MANDIBULAR PHEROMONE ON INITIATION AND MAINTENANCE OF QUEEN CELLS IN THE HONEY BEE (APIS MELLIFERA L.) , 1996, The Canadian Entomologist.

[47]  G. Robinson,et al.  Gene Expression Profiles in the Brain Predict Behavior in Individual Honey Bees , 2003, Science.

[48]  M. Wolfner,et al.  Seminal fluid regulation of female sexual attractiveness in Drosophila melanogaster. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Tautz,et al.  Beyond 9-ODA: SEX Pheromone Communication in the European Honey Bee Apis mellifera L. , 2006, Journal of Chemical Ecology.

[50]  D. Tarpy,et al.  Sampling Error, Effective Paternity, and Estimating the Genetic Structure of Honey Bee Colonies (Hymenoptera: Apidae) , 2002 .

[51]  D. Tarpy,et al.  Sex determination and the evolution of polyandry in honey bees (Apis mellifera) , 2002, Behavioral Ecology and Sociobiology.

[52]  G. Robinson,et al.  Influence of Gene Action Across Different Time Scales on Behavior , 2002, Science.

[53]  C. Butler,et al.  The Role of the Queen in Preventing Oogenesis in Worker Honeybees , 1963 .

[54]  T. Sugawara.  Oviposition behaviour of the cricket Teleogryllus commodus: The site of action of an oviposition-stimulating factor and the role of the nervous system , 1986 .

[55]  M. Wolfner,et al.  Comparative structural modeling and inference of conserved protein classes in Drosophila seminal fluid. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[56]  N. Gary,et al.  Chemical Mating Attractants in the Queen Honey Bee , 1962, Science.

[57]  G. Robinson,et al.  Volume Changes in the Mushroom Bodies of Adult Honey Bee Queens , 1995, Neurobiology of Learning and Memory.

[58]  T. Chapman,et al.  Functions and analysis of the seminal fluid proteins of male Drosophila melanogaster fruit flies , 2004, Peptides.

[59]  D. Tarpy,et al.  No Behavioral Control over Mating Frequency in Queen Honey Bees (Apis mellifera L.): Implications for the Evolution of Extreme Polyandry , 2000, The American Naturalist.

[60]  A. Łomnicki,et al.  Multiple mating of queens and the sterility of workers among eusocial hymenoptera , 1987 .

[61]  N. Koeniger.  The biology of the honey bee , 1988, Insectes Sociaux.

[62]  T. Seeley,et al.  Queen promiscuity lowers disease within honeybee colonies , 2007, Proceedings of the Royal Society B: Biological Sciences.

[63]  H. H. Laidlaw.  Instrumental Insemination of Honeybee Queens: Its Origin and Development , 1987 .

[64]  L. Riddiford,et al.  THE SWITCHOVER FROM VIRGIN TO MATED BEHAVIOR IN FEMALE CECROPIA MOTHS: THE ROLE OF THE BURSA COPULATRIX. , 1973, The Biological bulletin.

[65]  M. Wolfner,et al.  The Drosophila seminal fluid protein Acp26Aa stimulates release of oocytes by the ovary , 2000, Current Biology.

[66]  M. Winston,et al.  Honeybee response to queen mandibular pheromone in laboratory bioassays , 1990, Journal of Chemical Ecology.

[67]  W L Pak,et al.  cGMP-dependent changes in phototaxis: a possible role for the foraging gene in honey bee division of labor , 2003, Journal of Experimental Biology.

[68]  M. Wolfner,et al.  Tokens of love: functions and regulation of Drosophila male accessory gland products. , 1997, Insect biochemistry and molecular biology.

[69]  G. Robinson,et al.  Queen mandibular gland pheromone influences worker honey bee (Apis mellifera L.) foraging ontogeny and juvenile hormone titers. , 1998, Journal of insect physiology.