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Functions of microbial symbionts in fungus-farming beetles

Fruiting bodies of ambrosia fungus under SEM microscope. © Peter Biedermann
Fruiting bodies of a Raffaella ambrosia fungus under SEM microscope (1000× magnification). © Peter Biedermann
Mold fungus under SEM microscope. ©  Universitätsklinikum Jena
Mold fungus under SEM microscope.
© Universitätsklinikum Jena

Ambrosia beetles, like the fruittree pinhole borer Xyleborinus saxesenii, live within self-dwelled systems in wood. On the walls of the tunnels and chambers, they grow their fungal gardens. The gardens are dominated by two food fungi, but other microbes grow there as well. Some mold fungi seem to be parasitic, and they negatively affect the beetles’ offspring numbers. In this project, we aim at gaining insights into the factors influencing the composition of the microbial community in the beetles’ nests and the mechanisms of beetle fungiculture.

In a first step, we identify the community of microbes associated with two native ambrosia beetle species, the fruittree pinhole borer X. saxesenii and the black timber bark beetle Xylosandrus germanus. We investigate the succession in which different microbes occur within the nests. Subsequently, we test how these microbes interact with each other. To this end, we apply state-of-the-art molecular and visualizing techniques (i.e., next-generation sequencing, SEM, FISH). 

Previous studies have indicated that X. saxesenii beetles are able to influence the fungi growing in their nests in two ways: First, they are able to suppress the spread of molds. Second, they can enhance the growth and fruiting of their food fungus. Only in presence of beetles, the fungus produces fruiting bodies, so called conidia. The underlying mechanisms of these effects are also studied in this project. Preliminary results indicate that the beetles make use of other symbionts, which hence represent additional mutualistic player in this system, besides the beetles and their food fungus.

People involved in this project:

Pamela Baumann
(Bachelor and Master project)


Dan Vanderpool
Microbial genomics and symbiosis, Univ. of Montana, US

Miroslav Kolarik
Institute of Microbiology, Czech Academy of Science, Prague, CZ 

Symbionts of parasitic Dendroctonus bark beetles

Different symbionts cultivated in the lab. ©  Jürgen Lösel
Different symbionts cultivated in the lab.
© Jürgen Lösel

Bark beetles (Curculionidae: Scolytinae) of the genus Dendroctonus live in symbiosis with fungi and are widely used as model insects to study symbiosis. This is because they cover a wide range of ecological niches on the one hand, and on the other hand they have a huge economic impact. There are aggressive species that kill the trees that they inhabit, and others that inhabit stressed or dead trees. A third, parasitic group of Dendroctonus species develop in living trees without killing them. This third group is hardly studied, but very interesting as their hosts unlike dead trees are continuously defending themselves and it is unknown how the beetles overcome the tree defences.

Therefore, the beetles associate with peculiar bacterial and fungal microorganisms that help them to overcome the tree defenses. In this study, we use complementary culture-dependent and culture-independent methods to provide the first comprehensive overview of the bacterial and fungal symbionts of the parasitic beetles D. micans, D. punctatus, and D. valens. We also compare their microbial communities with those of other tree-inhabiting insects. Our first results show that many bacteria and yeasts are shared among the three species and their developmental stages, especially nitrogen-fixing and detoxifying microorganisms. Our study casts new light on the symbionts of bark beetles, and will contribute to understand how symbioses affect insects with various ecologies.  

People involved in this project:

Loic Dohet
(PhD project)
Biological Control and Spatial Ecology Lab (LUBIES), Université Libre de Bruxelles, BE


Jean-Claude Gregoire
Biological Control and Spatial Ecology Lab (LUBIES), Université Libre de Bruxelles, BE

Social immunity in ambrosia beetles & chemical recognition of fungal symbionts

Two X. saxesenii adults. © Gernot Kunz
Adults of X. saxesenii in a nest. © Gernot Kunz

In complex insect societies, the different individuals live in close spatial association and are highly related to each other. These two factors should favor the spread of parasites, but also the evolution of social behaviors to fight them, apart from chemical defense mechanisms. In the first part of this project, we study the social defense mechanisms in the ambrosia beetle Xyleborinus saxesenii. To this end, we experimentally expose them to pathogenic microorganisms, namely mold fungi. First results show that the beetles actively up-regulate hygienic behaviors like grooming each other (allo-grooming) and themselves (self-grooming), thereby likely reducing parasite load on their body surfaces, and cannibalism, thereby getting rid of infected individuals. Such collective pathogen defenses (social immunity) have been unknown in beetles so far.

In the second part of this project, we test if the beetles use olfactory cues to orient themselves during dispersal in order to locate a suitable habitat, i.e., a tree that has recently died and probably has specific chemical characteristics. In a pilot study, we found that the beetles are attracted to the volatiles of their main mutualistic fungus and to ethanol, which is emitted by recently dead trees due to degradation processes. This indicates that the beetles select new breeding grounds based on the presence of conspecifics that have already found a fungus garden successfully, and on the status of the host tree. Surprisingly, we did not find a repellent effect of volatiles produced by pathogenic fungi.

People involved in this project:

Jon Andreja Nuotclà
(MSc and PhD project)
Dep. Behavioural Ecology, University of Bern, CH


Prof Michael Taborsky
Dep. Behaviour Ecology, University of Bern, CH

Gerrit Holighaus, 
Dep Forest Zoology, University of Göttingen, DE

Dineshkumar Kandasamy,
Dep Biochemistry, MPI-CE, DE

Prof Marko Rohlfs
Dep. Population Ecology, University of Bremen, DE

Yeganeh Gharabigloozare, 
Dep. Biochemistry, MPI-CE, DE

Further reading:

Nuotclà (2015): Master thesis.
Available from author on request.

Diversity of fungal symbionts of Ambrosia beetles

Lab culture of ambrosia fungus. ©  Peter Biedermann
Lab culture of ambrosia fungus. © Peter Biedermann

Ambrosia beetles live in tunnel systems (‘galleries’) within the wood of recently dead or dying trees, where they plant a mixture of fungi on the tunnel walls. Exact compositions of these fungal communities are hardly studied, but it is known that they are dominated by beetle-specific ambrosia fungus species. Ambrosia fungi are Ascomycetes belonging to the genera Raffaelea (Sordariomycetidae), Ambrosiella (Ceratocystidaceae) or Fusarium (Hypocreales). They produce swollen hyphens, which serve as the beetles’ sole food source. It is known that the fungi reproduce asexually, but it has never been investigated if sexual reproduction also occurs. When females leave their natal nest and disperse, they carry asexual spores to their new nest, a mechanism called vertical transmission. However, it is also not fully clear if occasional transmission between existing nests (horizontal transmission) is possible.

If the fungi only reproduce asexually, and if vertical transmission is the only way spores reach new nests, the result would be low genetic diversity of fungi among beetle populations. In this project, we determine the genetic diversity of fungi from beetle populations all around Central Europe. The aim is to unravel the respective diversity of Ambrosiella or Raffaelea genotypes across and within beetle populations. Our results will help to understand how frequently horizontal exchange of symbiotic fungi occurs between nests, and they will shed light on the occurrence and frequency of sexual reproduction of the fungi. In addition, new fungi species will be identified in collaboration with T. Harrington.

People involved in this project:

Loic Dohet
(PhD project)
Biological Control and Spatial Ecology Lab (LUBIES), Université Libre de Bruxelles, BE


Prof Tom Harrington
Department of Plant Pathology, Iowa State University, US

Social behaviour of the coffee berry borer

A coffe berry borer on a coffee seed. ©  Peggy Grebb / USDA-ARS
A coffee berry borer on a coffee seed.
© Peggy Grebb / USDA-ARS

The coffee berry borer (Hypothenemus hampei) is the major pest of coffee, causing immense yield losses in coffee-producing countries worldwide. If we want to assess potential methods for biocontrol of this species, we need to better understand its life cycle and behavior, as well as its interaction with pathogens.

The beetle spends most of its life cycle inside the coffee berry. In order to elucidate the behavior of the coffee berry borer, we developed a diet sandwich, consisting of coffee berry borer artificial diet within two glass plates. This construction allows for observing the beetles inside their nests. We are currently determining the behaviors of the larvae, males, mated and unmated females. We are particularly curious if we find signs of a social lifestyle, namely delayed dispersal of adult offspring from their mother’s nest and their engagement in cooperative tasks and breeding. Further, we test how the beetles react towards an experimental injection of microbial pathogens (e.g., Beauveria bassiana) into their artificial diet in order to explore the potential of using natural enemies for biocontrol of this pest.

Principal investigator:

Fernando Vega,
US Department of Agriculture, Maryland, US

Interactions of native and invasive ambrosia beetles

Lab-reared ambrosia beetles cultures within glass tubes. ©  Peter Biedermann
Lab-reared ambrosia beetles cultures within glass tubes. © Peter Biedermann

International movement of non-native forest insects poses a severe threat to forests worldwide. Bark and ambrosia beetles (Curculionidae: Scolytinae) represent one of the most successful groups of invaders, in part because they are easily transported in wood products and packaging material. As international trade continues to steadily increase, the risks of introducing these organisms are also on the rise. Understanding why bark and ambrosia beetles establish so easily in habitats outside their native range is recognized as a research priority by both scientists and forest managers, with the aim to promote risk assessment and management strategies for non-native species.

In this project, we want to assess what effect non-native ambrosia beetle species can have on native ones. Therefore, we test the competitive abilities of a native versus a non-native ambrosia beetle by rearing them together in artificial media. Furthermore, to investigate if competitive effects are mediated by the symbiotic fungi of the beetles, we test the main fungal associate of the two beetle species against each other on agar plates.

Principal investigator:

Davide Rassati,
DAFNAE-Entomology, University of Padova, IT

Responses of fungal endophytes of black poplar to aphids

Pemphigus aphid gall on poplar. © S. Unsicker

Galling Pemphigus aphids induce major changes in the defense chemistry of black poplar. In this project we determine how these chemical changes affect the plants' fungal pathogen and endosymbiont communities.

We collect leaf samples with and without aphids in the field, determine their defense chemistry and culture fungi from these samples. In addition we check how major fungal players interact with each other and how they affect the leaf chemistry in experimental trials.


Sybille Unsicker,
Dep. Biochemistry, MPI-CE

Interactions of thistles, endosymbiotic fungi, and beetle herbivores

Thistle tortoise beetle (Cassida rubiginosa) © Andrew Cannizzaro / Wikipedia / CC BY 2.0
Thistle tortoise beetle (Cassida rubiginosa)© Andrew Cannizzaro / Wikipedia / CC BY 2.0

Like many other plants, thistles are often infected with endosymbiotic fungi that grow within their leaves. The plants are inoculated with the fungi when tortoise beetles feed on them, as they vector fungal spores.

In this project, we studied the community of endosymbiotic fungi in the cabbage thistle Cirsium oleraceum and the creeping thistle Cirsium arvense (also referred to as the Canada thistle). We were interested in wheter the beetles gain fitness benefits from feeding on fungus-infested leaves. To this end, we conducted feeding experiments using infected versus non-infected cabbage thistles and the thistle tortoise beetle Cassida rubiginosa.

People involved in this project:

Margo Wisselink,
Laboratory of Genetics, Wageningen University, NL


Prof Duur Aanen
Laboratory of Genetics, Wageningen University, NL