As she was preparing to do her dissertation research on the interactions between the Asian chestnut gall wasp, the chestnut blight disease and the European chestnut, Pilar Fernandez-Conradi read a lot of papers about fungal-insect-plant interactions.  She was impressed by the diversity of outcomes that resulted when plants were attacked by both insects and fungi, and wondered whether there were any generalities to glean from these research findings. She asked two basic questions. First, if a plant is infected by a fungus, is it more or less likely to be attacked by insects than is an uninfected plant?  Second, does an insect that attacks a fungal-infected plant perform better or worse than it would have on an uninfected plant?

Three-way interaction between the chestnut tree, the chestnut gall wasp, and the fungus Gnomopsis castanea. Female wasps induce the plant to create galls, which house developing larvae. Green globular galls (with a hint of rose-color) have not been infected by a fungus, while the very dark tissue is the the remains of a gall that was attacked by the fungus. Credit: Pilar Fernandez-Conradi.

Fernandez-Conradi and her colleagues thought they were more likely to discover a negative effect of fungal infection on the preference and performance of herbivorous insects.  Several studies had shown that nutrient quantity and quality of host plants is reduced by fungal infection, so it makes sense that insects would avoid infected plants.  But the researchers also knew that fungal infection can, in some cases, actually increase the sugar concentration of some plants, so insects might prefer those plants and also develop more rapidly on them. In addition, fungal infection can induce chemical defenses in plants that might make them less palatable to insects, or alternatively, fungal infection could weaken plant defenses making them more palatable to attacking insects.

To resolve this conundrum, Fernandez-Conradi and her colleagues did a meta-analysis, of the existing literature, identifying 1113 case studies based on 101 papers.  To be considered in the meta-analysis, all of the studies had to meet the following criteria: (1) report insect preference or performance on fungal-infected vs. uninfected plants, (2) report the Genus or species of the plant, fungus and insect, (3) report the mean response and a measure of variation (standard error, standard deviation or variance). The measure of variation allows researchers to calculate the effect size, which calculates the strength of the relationship that is being explored. The researchers found that, in general, insects avoid and perform worse on infected plants than they do on uninfected plants.

Mean effect size of insect preference and performance (combined) in response to fungal infection infection.  Error bars are 95% confidence intervals (CIs).  In this graph, and the next two graphs as well, a solid data point indicates a statistically significant effect.  You can also visually test for statistical significance by noting that the error bar does not cross the dashed vertical line that represents no effect (at the 0.0 value). The negative value indicates that insects respond negatively to fungal infection.

Fernandez-Conradi and her colleagues then broke down the data to explore several questions in more detail. For example, they wondered if the type of fungus mattered.  For their meta-analysis, they considered three types of fungi with different lifestyles: (1) biotrophic pathogens that develop on and extract nutrients from living plant tissues, (2) necrotrophic pathogens that secrete enzymes that kill plant cells, so they can develop and feed on the dead tissue, and (3) endophytes that live inside living plant tissue without causing visible disease symptoms.

Effect of fungus lifestyle on insect performance.  k = the number of studies.  Different letters to the right of CIs indicate significant differences among the variables (lifestyles).

The meta-analysis showed an important fungus-lifestyle effect (see the graph to your left).  Insect performance was strongly reduced in biotrophic pathogens and endophytes, but not in necrotrophic pathogens, where insect performance actually improved slightly (but not significantly). The researchers point out that biotrophic pathogens and endophytes both develop in living plant tissues, while necrotrophic pathogens release cell-wall degrading enzymes which can cause the plant to release sugars and other nutrients.  These nutrients obviously benefit the fungus, but can additionally benefit insects that feed on the plants.

To further explore this lifestyle effect, Fernandez-Conradi and her colleagues broke down insect response into performance and preference, focusing on chewing insects, for which there were the most data. Insects showed lower performance on and reduced preference (i.e. increased avoidance) for plants infected with biotrophic pathogens. They also performed equally poorly on endophyte-infected plants, but did not avoid endophyte-infected plants (see graph below). This was surprising since you would expect natural selection to favor insects that can choose the best plants to feed on. The problem for insects may be that endophytic infection is basically symptomless, so the insects may, in many cases, be unable to tell that the plant is infected, and likely to be less nutritionally rewarding.

Effects of fungal infection on preference and performance of chewing insects.  k = the number of studies.  Different letters to the right of CIs indicate significant differences among the variables. Variables that share one letter have similar effect sizes.

Many ecological studies deal with two interacting species: a predator and a prey, or a parasite and its host.  Fernandez-Conradi and her colleagues remind us that though two-species interactions are much easier to study, many important real-world interactions involve three or more species. Their meta-analysis highlights that plant infection by pathogenic and endophytic fungi reduces the performance and preference of insects that feed on these plants. But fungus lifestyle plays an important role, and may have different effects on performance and preference. Their meta-analysis also suggests other related avenues for research.  For example, how are plant-fungus-insect interactions modified by other species, such as viruses, bacteria and parasitoids (an animal that lives on or inside an insect, and feeds on its tissues)? Or, what are the underlying molecular (hormonal) mechanisms that determine the response of the plant to fungal infection, and to insect attack?  Finally, how does time influence both plant and insect response?  If a plant is recently infected by a fungus, does it have a different effect on insect performance and preference than does a plant that has suffered from chronic infection.  There are very few data on these (and other) questions, but they are more likely to get pursued now that some basic relationships have been uncovered.

note: the paper that describes this research is from the journal Ecology. The reference is Fernandez‐Conradi, P., Jactel, H., Robin, C., Tack, A.J. and Castagneyrol, B., 2018. Fungi reduce preference and performance of insect herbivores on challenged plants. Ecology, 99(2), pp.300-311. Thanks to the Ecological Society of America for allowing me to use figures from the paper. Copyright © 2018 by the Ecological Society of America. All rights reserved.