Helge Schlüns

Multiple nuptial flights, sperm transfer and the evolution of extreme polyandry in honeybee queens

The honeybee, Apis mellifera, has an extremely polyandrous mating system, which often involves multiple nuptial flights by its queens. To understand the evolution of extreme polyandry, we investigated the cost of multiple nuptial flights in relation to potential benefits. We analysed, with eight DNA microsatellite loci, the paternity of worker offspring of naturally mated queens. Queens that were restricted to one nuptial flight, but may have taken additional nuptial flights if allowed to do so, had significantly fewer matings than queens that started oviposition after a single nuptial flight. Furthermore, the number of sperm stored in a spermatheca increased significantly with the number of matings. We suggest that queens adjust their nuptial flight frequencies according to their mating success in their previous nuptial flights. The number of copulations seems to serve as a signal for the initiation of oviposition. In the light of these findings, we reconsider and discuss the significance of the sperm limitation hypothesis for the evolution of extreme polyandry in A. mellifera.

Relish regulates expression of antimicrobial peptide genes in the honeybee, Apis mellifera, shown by RNA interference

Relationships of immune genes in adult honeybees (Apis mellifera) were investigated using RNA interference (RNAi). Quantitative RT‐PCR was applied to estimate gene expression and the extent of gene silencing. Relish is a transcription factor and forms an important part of the IMD signalling pathway. The expression of the immune gene Relish was significantly reduced by RNAi (ca. 70%). The proposed regulation of antimicrobial peptide genes by Relish could be established for abaecin and hymenoptaecin. These two genes showed a reduction in gene expression to the same extent as Relish. However, the antimicrobial peptide gene defensin‐1 was not affected which suggests defensin‐1 is regulated by a different signalling pathway.

Infection with the trypanosome Crithidia bombi and expression of immune-related genes in the bumblebee Bombus terrestris

Social bees and other insects are frequently parasitized by a large range of different microorganisms. Among these is Crithidia bombi (Kinetoplastida: Trypanosomatidae), a common gut parasite of bumblebees, Bombus spp. (Insecta: Apidae). Bumblebees are important pollinators in commercial and natural environments. There are clear detrimental effects of C. bombi infections on the fitness of bumblebees. However, little has been known about how the bees immune system responds to infections with trypanosome parasites. Here, we study the immune response of Bombus terrestris on infection by C. bombi. We measured the expression of four immune-related genes (Hemomucin, MyD88, Relish, and TEP7) using RT-qPCR in adult B. terrestris workers that were either healthy or infected with the trypanosome parasite C. bombi. The potential recognition gene Hemomucin was significantly upregulated in the infected bees. Further, there was substantial and significant variation in all four genes among different bumblebee colonies irrespective of infection status.

Breeding system, colony and population structure in the weaver ant Oecophylla smaragdina

Weaver ants (Oecophylla smaragdina) are dominant ants in open forests from India, Australia, China and Southeast Asia, whose leaf nests are held together with larval silk. The species, together with its sole congener O. longinoda, has been important in research on biological control, communication, territoriality and colony integration. Over most of the range, only one queen has been found per colony, but the occurrence of several queens per nest has been reported for the Australian Northern Territory. The number of males mating with each queen is little known. Here we report on the colony structure of O. smaragdina using published and new microsatellite markers. Worker genotype arrays reflect the occurrence of habitual polygyny (more than one queen per colony) in 18 colonies from Darwin, Northern Australia, with up to five queens inferred per colony. Monogyny (one queen per colony) with occasional polygyny was inferred for 14 colonies from Queensland, Australia, and 20 colonies from Java, Indonesia. Direct genotyping of the sperm carried by 77 Queensland queens and worker genotypic arrays of established colonies yielded similar results, indicating that less than half of the queens mate only once and some mate up to five times. Worker genotype arrays indicated that queens from Java and the Northern Territory also often mate with more than one male, but less often than those from Queensland. A strong isolation‐by‐distance effect was found for Queensland samples. The variation uncovered means that O. smaragdina is a more versatile study system than previously supposed.

Intraspecific support for the polygyny-vs.-polyandry hypothesis in the bulldog ant Myrmecia brevinoda

The number of queens per colony and the number of matings per queen are the most important determinants of the genetic structure of ant colonies, and understanding their interrelationship is essential to the study of social evolution. The polygyny‐vs.‐polyandry hypothesis argues that polygyny and polyandry should be negatively associated because both can result in increased intracolonial genetic variability and have costs. However, evidence for this long‐debated hypothesis has been lacking at the intraspecific level. Here, we investigated the colony genetic structure in the Australian bulldog ant Myrmecia brevinoda. The numbers of queens per colony varied from 1 to 6. Nestmate queens within polygynous colonies were on average related (rqq = 0.171 ± 0.019), but the overall relatedness between queens and their mates was indistinguishable from zero (rqm = 0.037 ± 0.030). Queens were inferred to mate with 1–10 males. A lack of genetic isolation by distance among nests indicated the prevalence of independent colony foundation. In accordance with the polygyny‐vs.‐polyandry hypothesis, the number of queens per colony was significantly negatively associated with the estimated number of matings (Spearman rank correlation R = −0.490, P = 0.028). This study thus provides the rare intraspecific evidence for the polygyny‐vs.‐polyandry hypothesis. We suggest that the high costs of multiple matings and the strong effect of multiple mating on intracolonial genetic diversity may be essential to the negative association between polygyny and polyandry and that any attempt to empirically test this hypothesis should place emphasis upon these two key underlying aspects.

Genetic caste determination in termites: out of the shade but not from Mars.

Several ant species are known with genetic effects on caste determination but, for termites, the role of environment has been assumed to be omnipotent. Now Hayashi et al. report that commitment to the nymph and worker pathways in Reticulitermes speratus follows a simple model involving two alleles at a sex-linked locus. The spread of this system of genetic caste determination seems best explained by selection at the colony level. This remarkable system may be widely applicable throughout termites, although it cannot be universal, and may provide a window into causal aspects of the molecular biology of caste determination.

Characterization of Polymorphic Microsatellites in the Giant Bulldog Ant, Myrmecia brevinoda and the Jumper Ant, M. pilosula

Abstract The ant genus Myrmecia Fabricius (Hymenoptera: Formicidae) is endemic to Australia and New Caledonia, and has retained many biological traits that are considered to be basal in the family Formicidae. Here, a set of 16 dinucleotide microsatellite loci were studied that are polymorphic in at least one of the two species of the genus: the giant bulldog ant, M. brevinoda Forel, and the jumper ant, M. pilosula Smith; 13 are novel loci and 3 are loci previously published for the genus Nothomyrmecia Clark (Hymenoptera: Formicidae). In M. brevinoda, the total of 12 polymorphic microsatellites yielded a total of 125 alleles, ranging from 3 to 18 with an average of 10.42 per locus; the observed and expected heterozygosities ranged from 0.4000 to 0.9000 and from 0.5413 to 0.9200, respectively. In M. pilosula, the 9 polymorphic loci yielded a total of 67 alleles, ranging from 3 to 12 with an average of 7.44 per locus; the observed and expected heterozygosities ranged from 0.5625 to 0.9375 and from 0.4863 to 0.8711, respectively. Five loci were polymorphic in both target species. In addition, 15 out of the 16 loci were successfully amplified in M. pyriformis. These informative microsatellite loci provide a powerful tool for investigating the population and colony genetic structure of M. brevinoda and M. pilosula, and may also be applicable to a range of congeners considering the relatively distant phylogenetic relatedness between M. pilosula and the other two species within the genus Myrmecia.

Colony genetic structure in the Australian jumper ant Myrmecia pilosula

Eusocial insects vary significantly in colony queen number and mating frequency, resulting in a wide range of social structures. Detailed studies of colony genetic structure are essential to elucidate how various factors affect the relatedness and the sociogenetic organization of colonies. In this study, we investigated the colony structure of the Australian jumper ant Myrmecia pilosula using polymorphic microsatellite markers. Nestmate queens within polygynous colonies, and queens and their mates, were generally unrelated. The number of queens per colony ranged from 1 to 4. Queens were estimated to mate with 1–9 inferred and 1.0–11.4 effective mates. This is the first time that the rare co-occurrence of polygyny and high polyandry has been found in the M. pilosula species group. Significant maternity and paternity skews were detected at the population level. We also found an isolation-by-distance pattern, and together with the occurrence of polygynous polydomy, this suggests the occurrence of dependent colony foundation in M. pilosula; however, independent colony foundation may co-occur since queens of this species have fully developed wings and can fly. There is no support for the predicted negative association between polygyny and polyandry in ants.