Too much of a good thing is killing Monarch butterflies

There was a time in the mid-Pleisticine when a photo of an ecological event was an awesome novelty, and a movie of an ecological event even more so.  Dodderers of an ecological bent (myself included), can vividly recall viewing a series of photos or a movie, either in a seminar or in an ancient ecology text, of a blue jay consuming a monarch butterfly, Danaus plexippus.  Consumption is immediately followed by explosive vomiting, as the cardenolides within the monarch butterfly claim another victim.  The monarch sequesters these cardenolide toxins from its larval food (milkweed), and incorporates them into its tissues as a means of protecting itself from predators – presumably blue jays learn from this very aversive experience.  I should point out that the individual sacrificial butterfly enjoys no fitness from this learning event – which raises some evolutionary questions we will not explore at the present.

Karen Oberhauser

Five instars (stages of development) of monarch caterpillars on a milkweed leaf. Credit: Karen Oberhauser

Rather we turn our attention to the relationship between milkweed, monarchs, and climate change. In several places in this blog we’ve talked about how climate change has influenced the behavior or physiology of a single species. For example, my first blog (Jan 31, 2017) discusses how increasing temperatures create more females in a loggerhead turtle population. But there are fewer studies that explore how climate change influences the ecological landscape, ultimately affecting interactions between species.  Along these lines, Matt Faldyn wondered if increased air temperature would change the chemical constitution of milkweed in a way that might influence monarch populations.  As he describes, “With milkweed toxicity, there is a ‘goldilocks’ zone where monarchs prefer to feed on milkweed that produce enough toxins in order to sequester these (cardenolide) chemicals as an antipredator/antiparasite defense, while also avoiding reaching a tipping point of toxicity where feeding on very toxic milkweeds negatively impacts monarch fitness.” He expected that at higher temperatures, milkweed would become stressed, and be physiologically unable to sustain normal levels of cardenolide production.

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Monarch butterfly feeds on a native milkweed, Asclepias incarnata. Credit: Teune at the English Language Wikipedia.

For their research, Faldyn and his colleagues worked with two milkweed species.  Asclepias incarnata is a common, native milkweed found throughout the monarch butterfly’s range in the eastern and southeastern United States.  Asclepias curassavica is an exotic species that has become established in the southern United States.  In contrast to A. incarnata, A. curassavica does not die back over the winter months; consequently some monarch populations are no longer migratory, relying on A. curassavicato provide them with a year round food supply.

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The exotic milkweed, Asclepias curassavica. Credit: 2016 Jee & Rani Nature Photography (License: CC BY-SA 4.0)

To protect against herbivory, milkweeds have two primary chemical deterrants: (1) the already-mentioned cardenolides, which are toxic steroids that disrupt cell membrane function, and (2) release of sticky latex, which can gum up caterpillar mouthparts and actually trap young caterpillars.

field_noborderii.jpgThe researchers wanted to simulate climate change under field conditions, so they created open-top chambers with plexiglass plates that functioned much like mini-greenhouses, into which they placed one milkweed plant that was covered with butterfly netting.  This setup raised ambient temperatures by about 3°C during the day and 0.2°C at nighttime.  Control plots were single milkweed plants with butterfly netting. Half of the plants were native milkweed, and the other half were the exotic species.

For their experiments, Faldyn and his colleagues introduced 80 monarch caterpillars (one per plant) and allowed them to feed normally until they pupated.  Pupae were brought into the lab and allowed to metamorphose into adults.

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Matt Faldyn holds two monarch butterflies in the laboratory. Credit Matt Faldyn.

At normal (ambient) temperatures, monarchs survived somewhat better on exotic milkweed.  But at warmer temperatures, there is a strikingly different picture. Monarch survival is unaffected by warmer temperatures on native milkweed, but is sharply reduced by warmer temperatures on exotic milkweed (top graph below). The few that managed to survive warm temperatures on exotic milkweed grew much smaller, based on their body mass and forewing length (middle and bottom graph below)

FaldynFig1

Survival (top), adult mass (middle) and forewing length (bottom) of monarch butterflies raised under normal (ambient) and warmed temperatures.  Error bars are 95% confidence intervals.

Both milkweed species increased production of both types of chemicals over the course of the experiment. But by the end of the experiment, the exotic species released 3-times the quantity of latex and 13-times the quantity of cardenolides than did the native milkweed species.

FaldynFig2

Average amount of latex released at the beginning and end of the experiment.  Error bars are 95% confidence intervals.

FaldynFig2

Average cardenolide concentration at the beginning and end of the experiment.

The researchers argue that the exotic milkweed, Asclepias curassavica, may become an ecological trap for monarch butterflies, in that it attracts monarchs to feed on it, but will, under future warmer conditions, result in dramatically reduced monarch survival. Interestingly, these results are not what Faldyn originally expected; recall that he anticipated that temperature-stressed plants would reduce cardenolide production. The tremendous increase in cardenolide production in exotic milkweed at warmer temperatures may simply be too much toxin for the monarchs to process. The researchers predict that as climate warms, milkweed ranges will expand further north into Canada, and lead to northward shifts of monarch populations as well.  They urge nurseries to emphasize the distribution of native rather than exotic milkweed, so that monarchs will be less likely to become victims of this ecological trap.

note: the paper that describes this research is from the journal Ecology. The reference is Faldyn, M. J., Hunter, M. D. and Elderd, B. D. (2018), Climate change and an invasive, tropical milkweed: an ecological trap for monarch butterflies. Ecology. doi:10.1002/ecy.2198. 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.

Fungi attack plants – insects respond!

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?

D. Kuriphilus+Gnomo

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.

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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.

Fernandez-conradiFig1

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.

Fernandez-conradiFig2

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.

Parrotfish put on their big boy pants

While it would be awesome if parrotfish were named for their conversational abilities, it turns out that they earn their moniker for their specialized teeth that are fused together for scraping algae from coral, thus resembling a parrot’s beak. Despite lacking verbal skills these fish are incredible. Approximately 100 species occupy reefs, rocky coastlines and eelgrass meadows in tropical and subtropical waters. Many species are sequential hermaphrodites, beginning life as females and then changing into males after reaching a certain size. While female reproductive success is limited by the number of eggs she can produce, male reproductive success can be much higher if he can fertilize the eggs of many females.  So if a parrotfish transitions into a large male, and can control access to numerous females, he will enjoy greater reproductive success than if he had remained a female.

C. spilurusBrettTaylor

Two Chlorurus spilurus parrotfish show off their teeth and colors.  The large colorful fish on the right is a male, while the smaller darker fish to his left is a female. Credit: Brett Taylor.

Phenotypic plasticity describes the ability of an individual with a particular genetic makeup to vary in a variety of traits (such as what it looks like, or how it behaves) in response to different environmental conditions. About 15 years ago, Nick Gust’s PhD research on tropical reef fish revealed that tremendous variation in parrotfish traits existed over a distance of a few kilometers. But what causes this variation? When funding became available, Brett Taylor jumped at the opportunity to pinpoint the causes, focusing on the diverse parrotfish community in the Great Barrier Reef (GBR).

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Eastern slope of the Great Barrier Reef hosts a diversity of fish and coral species. Credit: Brett Taylor.

Taylor and his colleagues surveyed 82 sites within 31 reefs across 6 degrees of latitude in the northern GBR. To standardize data collection, divers, armed with a multitude of cameras and GPS devices, swam at a standardized rate (about 20 meters/minute) for 40 minutes per survey, recording each parrotfish along a 5 m wide swath. They collected data about the habitat and the environment, about the physical traits of each individual parrotfish (such as size and sex), and about the type and abundance of parrotfish and their predators present at each site.

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Researcher takes notes while conducting a dive.  Credit Kendra Taylor.

The researchers wanted to identify what factors influenced growth rate, maximum body size, and the size at sex change, and how these factors related to the parrotfish mating system. Four species of parrotfish were sufficiently abundant across the GBR to allow researchers to do this type of analysis.

TaylorFig1

Four parrotfish species  abundant along exposed outer shelf (yellow sites) and protected inner shelf (blue) regions of the Great Barrier Reef. Males are larger and more colorful.

The GBR varies structurally across a relatively small spatial scale of 40 – 100 km, with outer shelf regions (eastern) exposed to wave action, and inner shelf regions (western) relatively protected. All four species tended to change sex at a larger size in protected sites than they did at exposed sites. However, the differences are only compelling for two of the species: C. spilurus and S. frenatus. There were fewer data points for the other two species, so it is possible (but unknown) that they too would show a more pronounced trend if more data were available.

TaylorFig1bottom

Proportion terminal phase (sex-changed males) in relation to body size (measured to the fork of the tail) in exposed (yellow) and sheltered (blue) sites.

Not surprisingly, parrotfish grew larger in protected areas. Presumably, less wave action provided a more benign environment for rapid growth, both of parrotfish and their preferred food items (algae growing on rocks and coral).

TaylorFig2I

Standardized maximum size (Lmax) attained by parrotfish in sheltered vs. exposed sites.

The researchers were somewhat surprised that most other factors, such as latitude, coral cover, sea surface temperature, and predator abundance, had very little effect on the size at sex change. Rather, the size at sex change appears to be strongly influenced by the local size distribution. In protected habitats, parrotfish grow large and change sex at a large size, while in exposed habitats, parrotfish are smaller, and change sex at a smaller size.

But sex is never simple. Nick Gust’s PhD research showed that C. spilurus had different patterns of sexual allocation in protected vs. exposed areas. In protected areas, the mating system is haremic, with a large male defending a territory and servicing a harem of females. In exposed areas, the mating system is mixed; there still are large territorial males with their harems, but they compete with many more small males, and group spawning is much more prevalent. Theoretically, the presence of these small males may make it less worthwhile for a female to transition into a male, and may influence the optimal size for transitioning in exposed reefs. Given that we still don’t know the mating system details of the other parrotfish in this study, it will be fascinating to see if they too show similar patterns of haremic vs. mixed mating systems in relation to habitat structure.

note: the paper that describes this research is from the journal Ecology. The reference is Taylor, B. M., Brandl, S. J., Kapur, M., Robbins, W. D., Johnson, G., Huveneers, C., Renaud, P. and Choat, J. H. (2018), Bottom-up processes mediated by social systems drive demographic traits of coral-reef fishes. Ecology 99(3): 642-651. doi:10.1002/ecy.2127. 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.

Field gentian – when it’s good to be eaten

We tend to think of plants as victims – after all any interested herbivore can simply walk, fly or crawl over to its favorite plant, and begin munching. But not so fast! In reality, plants have a variety of ways they can make life difficult for potential herbivores. Plants can escape herbivores by simply growing in places that are not easily accessible (such as in cracks, or high enough to be out of a herbivore’s reach) or by growing at a time of year when herbivores are away from the plant’s habitat. Plants also use mechanical defenses such as thorns or a diverse array of chemical defenses to thwart overzealous herbivores. A third approach – tolerance – can take many forms. For example, following attack by a herbivore some plants can increase photosynthetic rates or reduce the time until seed production . Tommy Lennartsson and his colleagues were interested in a particular form of tolerance that ecologists call overcompensation, in which damaged plants produce more seeds than undamaged plants.

LennartssonFigure1

Herbivores in action. Notice the difference in vegetation height inside and outside the pasture. Credit: Tommy Lennartsson.

Overcompensation is an evolutionary puzzle, because undisturbed plants produce fewer offspring than partially eaten plants. That outcome seems to fly in the face of the scientific principle that natural selection favors individuals with traits that promote reproductive success. Lennartsson and his colleagues investigated this evolutionary puzzle by comparing two subspecies of the herbaceous field gentian Gentianella campestris. The first subspecies, Gentianella campestris campestris (which we’ll just call campestris), has relatively unbranched shoot architecture when intact, growing to about 20 cm tall, but produces multiple fruiting branches when the dominant apical meristem is eaten. The second subspecies, Gentianella campestris islandica (which we’ll call islandica), is much shorter (about 5-10 cm tall), and always has a multi-branched architecture.

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Two subspecies of field gentian – campestris (left) and islandica (right).

Environmental conditions and soils can vary dramatically, even on a small spatial scale. The field site was a gently-sloped grassland in Sweden that had coarser, dryer soil on the ridge, and finer, wetter and richer soil in the valley. This created a productivity gradient, with taller vegetation in the valley. The average  height of all the vegetation was 15 cm in the high-productivity valley, 10 cm on the medium-productivity slope and 5 cm on the low-productivity ridge.

The researchers used this natural variation to set up an experiment that would allow them to explore hypotheses about why an undisturbed campestris is less successful than one that is partially-eaten. One hypothesis (the overcompensation hypothesis) is that campestris restrains branching to conserve resources, so that when it is grazed it has plenty of resources in reserve to be used for regrowth and the production of prolific branches, flowers and seeds. Limited branching and limited seed production of ungrazed campestris are simply a cost of tolerance, while overcompensation after damage maximizes reproductive success. A second hypothesis (the competition hypothesis) is that restrained branching allows the plant to grow tall, so it can compete better in ungrazed pastures than can the much shorter islandica. These two hypotheses are not mutually exclusive.

To test these two hypotheses, the researchers set up 2 X 2 meter experimental plots in the valley (18 plots), slope (12 plots) and the ridge (6 plots). They planted 2000 seeds per subspecies in each plot, which ultimately yielded about 20 plants of each subspecies per plot. Of course there were many other neighboring plant species in these plots. In the high productivity plots (valley), the neighboring plants in six plots were clipped to a height of 12 cm, six plots to 8 cm and six plots to 4 cm. In the medium productivity plots (which naturally only grew to 10 cm), the researchers cut neighboring plants to 8 cm in 6 plots and 4 cm in six plots. Finally, in the low productivity plots, the researchers cut neighboring plants to 4 cm in all six plots. In mid July, half of the gentian plants in each plot were clipped to the same height as the surrounding vegetation, while the remainder were not clipped.

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Experimental plots from the valley (left), slope (middle) and ridge (right).  Black squares represent plots where neighboring plants were clipped to 12 cm, grey squares to 8 cm, and clear squares to 4 cm. Squares with slashes through them (left)  represent plots that were used for a different purpose.

The beauty of this experimental design, is that by counting seeds, the researchers could assess the reproductive success of both subspecies under conditions of high competition (when surrounded by tall neighbors) and low competition (when surrounded by shorter neighbors). At the same time, clipping the two subspecies allowed the researchers to simulate grazing in these different competitive environments. Lennartsson and his colleagues found that unclipped islandica did better than unclipped campestris when surrounded by short or medium height neighbors, but that islandica success plummeted when the neighbors were very tall (see the left graph below). Campestris reproductive success also dropped when surrounded by tall competitors, but not as much as did islandica, so that campestris produced twice as many seeds than islandica in the high competition environment (also the left graph).

When plants were clipped to simulate grazing, campestris outperformed islandica in all three competitive environments. Campestris actually produced more seeds when it was clipped than when it was not clipped in the low and medium competition environments. Thus campestris overcompensated for grazing under conditions of low and moderate competition (see the right graph below).

LennartssonFig2

Mean (+ standard error) seed production for unclipped (left graph) and clipped (right graph) field gentian subspecies in relation to surrounding vegetation height.  Sample sizes are in bars.

The researchers collected data on growth rates, development, survival probabilities and reproductive success for both species under conditions of being clipped or unclipped at different levels of competition. They then used these data to create a population growth model in relation to the percentage of grazing (damage risk) at different levels of productivity. In these graphs, a stochastic growth rate of 1.0 (on the y-axis) indicates that the population is stable, above 1.0 indicates it will increase and below 1.0 indicates a declining population.

LennartssonFig4

Population growth rate of both subspecies in relation to damage risk at different levels of productivity.  These models predict that the population will increase at growth rates above the dotted line (growth rate = 1.0) and decline below the dotted line.

This model shows that in high productivity environments, campestris always does better than islandica (top graph). However, the model predicts that islandica will decline at any damage level (note in the top graph that all islandica damage values yield a growth rate below 1.0), while campestris will also decline except for very high damage risks. In medium and low productivity populations (middle and bottom graphs), islandica does better than campestris when damage risk is low, but the reverse is true at high damage risk.

So how do these results relate to the two hypotheses for why an undisturbed campestris is less successful than one that is partially-eaten. Campestris overcompensated for damage by producing more seeds and having positive population growth under most levels of productivity. In contrast, islandica undercompensated when damaged, but produced more seeds than campestris when ungrazed, except for in the high productivity environment. These differences in responses support the hypothesis that restrained branching is favored by natural selection in environments where damage from grazing is common (the overcompensation hypothesis). But, the superior performance by campestris in productive ungrazed environments supports the competition hypothesis.

Can we generalize these findings to other plants? Lennartsson and his colleagues point out that many short-lived grassland plants can’t grow tall enough to be effective competitors for light. These plants are thus restricted to environments where the surrounding plants are not very tall. Two factors commonly create conditions where there are short neighboring plants: grazing and unproductive (low nutrient) soils. When grazing is widespread, tolerance mechanisms such as overcompensation are favored by natural selection. When soils are unproductive, unrestrained branching is favored. Therefore, Gentianella campestris provides us with a natural experiment for testing hypotheses about how natural selection acts on plants to promote their reproductive success in a variable environment.

note: the paper that describes this research is from the journal Ecology. The reference is Lennartsson, T., Ramula, S. and Tuomi, J. (2018), Growing competitive or tolerant? Significance of apical dominance in the overcompensating herb Gentianella campestris. Ecology, 99: 259–269. doi:10.1002/ecy.2101. 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.

 

Powdery parasites pursue pedunculate oak

Studying disease transmission is tricky for many reasons. Most humans frown on what might seem like the easiest experimental protocol – release a disease into the environment and watch to see how it spreads. For his doctoral dissertation in 2006, Ayco Tack settled on a different experimental protocol – bring the potential hosts to the disease. In this study, staged in Finland, the hosts were pedunculate oak trees, Quercus robur, and the disease was the powdery mildew parasite, Erysiphe alphitoides. Almost 10 years later, Adam Ekholm continued research on the same system, with Tack as his co-supervisor.

Ayco Tack

Trees on the move. Credit: Ayco Tack.

But before moving trees around, the researchers first needed to see how the disease moved around under field conditions.  Within a tree stand, powdery mildew success will depend on how many trees it occupies, how many trees it colonizes in the future, and how many trees it disappears from (extinction rate). The researchers measured these rates over a four year period (2003 – 2006) on 1868 oak trees situated on the island of Wattkast in southwest Finland. They also measured spatial connectivity of each tree to others in the stand. In this case connectivity is a measure of the distance between a tree and other trees, weighted by the size of the other trees. So a tree that has many large neighbors nearby has high connectivity, while a tree with a few distant and mostly small neighbors has low connectivity. Results varied from year-to-year, but in general, the researchers found higher infection rates, lower extinction rates, and some evidence of higher colonization rates in trees with high connectivity.

Mildew_Adam Ekholm

Oak leaf infected with powdery mildew parasite. Credit: Adam Ekholm.

The importance of connectivity indicated that the parasites simply could not disperse efficiently to distant trees. But perhaps the environment might play a role in colonization rates as well. For example, fungi like powdery mildew tend to thrive in shady and humid environments. Thus a tree out in the open might resist colonization by powdery mildew more effectively than would a tree deep in the forest. To test this hypothesis, Tack and his colleagues placed 70 trees varying distances (up to 300 meters) from an infected oak stand. On one side of the oak stand was an open field, while the other side was closed forest. Thus two variables, distance and environment, could be investigated simultaneously.

Ayco Tack inspecting a potted tree_Tomas Roslin

Ayco Tack inspects an oak tree placed in an open field. Credit: Tomas Roslin.

The researchers collected infection data twice; once in the middle of the growing season (July) and a second time at the end of the growing season (September). Not surprisingly, infection rates were higher by the end of the growing season. In general, infection rates, and infection intensity (mildew abundance) were higher in the forest than in the field, indicating a strong environment effect. In the July survey, trees further from the oak stand had lower infection intensity, but as infection rates increased over the course of the season, the effects of distance diminished, particularly in the forest.

EkholmFig3

Upper two graphs show the impact of habitat type on (a) proportion of trees infected and (b) mildew abundance. The lower two graphs are the influence of distance from parasite source on mildew abundance of trees set in (c) a forest habitat and (d) an open field. Mildew abundance was scored on an ordinal scale with 0 = none and 4 = very abundant.

Ten years later, Adam Ekholm, as part of his PhD dissertation that studies the effect of climate on the insect community on oak trees, added a third element to the mix – the influence of genes on disease resistance. He wondered whether certain genotypes were more resistant to powdery mildew infection. The researchers grafted twigs from 12 large “mother” trees, creating 12 groups of trees, with between 2 – 27 trees per group (depending on grafting success). Each tree in a given group was thus genetically identical to all other trees within that group.

Ayco Tack

Oak tree placed in the forest. Credit: Ayco Tack.

The researchers chose a site that contained a dense stand of infected oaks, but was surrounded by a grassy matrix that contained only an occasional tree. To study the impact of early season exposure, Ekholm and his colleagues divided the trees into two groups; 128 trees were placed in the matrix at varying distances from the infected stand, while 58 trees were placed directly in the midst of the stand for about 50 days, and then moved varying distances away. The researchers scored trees for infection at the end of the growing season (mid-September).

 

Trees that spent 50 days within the oak stand had much higher infection frequency and intensity than trees that were initially placed in the matrix. Some genotypes (for example genotype I in graphs C and D below) were much more resistant to infection than others (such as genotypes D and J). Finally trees further from the source of infection were less susceptible to become colonized over the course of the summer (data not shown).

EkholmFig2

Proportion of trees infected (A) and proportion of leaves infected (B) in response to early season exposure to stand of oaks infected with the powdery mildew parasite (oak stand) or no early season exposure (matrix). Proportion of trees infected (C) and proportion of leaves infected (D) in relation to tree genotype. Genotypes are labeled A – L; numbers in parenthesis are sample size for each group.

These findings illustrate how dispersal, host genotype and the environment influence the spread of a parasite under natural conditions. The parasite exists as a metapopulation – a group of local populations inhabiting networks of somewhat discrete habitat patches. Some populations go extinct while others successfully colonize each year, depending on distance from a source, tree genotype and environment. Ekholm and his colleagues encourage researchers to use similar experimental approaches in other host-parasite systems to evaluate how general these findings are, and to explore how multiple factors interact to shape the dynamics of disease transmission.

note: the paper that describes this research is from the journal Ecology. The reference is Ekholm, Adam; Roslin, Tomas; Pulkkinen, Pertti and Tack, Ayco. J. M. (2017). Dispersal, host genotype and environment shape the spatial dynamics of a parasite in the wild. Ecology. doi:10.1002/ecy.1949. The paper should come out in print very soon. Meanwhile you can also link to Dr. Tack’s website at www.plantmicrobeinsect.com Thanks to the Ecological Society of America for allowing me to use figures from the paper. Copyright © 2017 by the Ecological Society of America. All rights reserved.

Spotted salamander performance per polymorphism persistence

His first winter at the University of Mississippi Field Station, Matt Pintar was wading through some ponds where he noticed a large number of egg masses. Clear jelly surrounded most of these egg masses, but a whitish jelly encased some of them. These egg masses were produced by the spotted salamander, Abystoma maculatum, which immediately made Pintar wonder why these differences exist within this species. Biologists use the term “polymorphism” to describe a situation like this, in which two or more forms (poly = multiple, morph = form) exist within a population.

Pintarmorphs

White egg mass (left) and clear egg mass (right). Credit: Matt Pintar

Could it simply be random chance that there were two egg mass morphs? Or was one morph better than the other in getting fertilized by the appropriate sperm, or in keeping the eggs together? Alternatively, perhaps one morph was better at providing nutrients or protecting against predators. The puzzle is that if one morph was superior to the other, then that morph would be favored by natural selection, should outcompete the other, and ultimately cause the second morph to go extinct. So why did both morphs persist in this population of spotted salamanders?

Pintaradult

Adult male spotted salamander. Credit Matt Pintar

Pintar embryos

Recently hatched larvae. Credit Matt Pintar.

Pintar and his colleague Willliam Resetarits Jr. thought it most likely that the polymorphism was a chance event that provided no benefit to the salamanders. But they did consider the alternative that one morph might be better in some conditions, while the other morph was better in other conditions. Surveys done about 25 years ago suggested that the polymorphism might be connected to differences in water chemistry, so Pintar and Resetarits decided to explore this possible link with a combination of observations of natural ponds and field experiments on artificial ponds.

Pintar ponds

Ponds at the University of Mississippi Field Station. Credit Matt Pintar.

Nutrient levels of ponds at the field station are influenced by two major factors: the type of surrounding habitat and the duration of time each pond holds water over the course of the year (the hydroperiod). Ponds surrounded by trees, as opposed to grass, have higher nutrient levels courtesy of tree leaves that fall into the ponds and leach out their nutrients. Many of these ponds dry out in the summer, so ponds with a longer hydroperiod will have more time to receive and leach nutrients from organic matter.

Ecologists often use water conductivity as a general measure of pond nutrient levels. Ponds with high conductivity have higher nutrient levels than ponds with low conductivity. Pintar and Resetarits sampled the water from 55 ponds and counted the number of white and clear egg masses from 40 ponds that had egg masses in 2015 and 2016. They found a striking relationship between conductivity and egg mass morph. White egg masses were much more common in low-nutrient (low conductivity) ponds.

Fig2A

Proportion of white egg masses in relation to water conductivity (a measure of nutrient level)

The researchers further explored this relationship by setting up artificial ponds that contained either low or high nutrient levels (obtained by putting leaf litter into some of the pools), and inoculating each pond with both white and clear egg masses. Pintar and Resetarits made sure that larvae were not mixed up once they hatched. They then measured the effects of both nutrient levels and morph on many variables associated with growth and development in these artificial ponds.

pintarmesocosm

Some of the artificial ponds used for controlled experiments on the effects of nutrient levels. Credit: Matt Pintar.

In general, eggs from white masses had a significantly higher rate of hatching (about 80%) at both nutrient levels than did eggs from clear masses (about 60%). But eggs from white masses took longer to hatch (Figure (c) below). Importantly, larvae from white masses tended to grow better under low-nutrient conditions than did larvae from clear masses. In contrast, larvae from clear masses grew better under high-nutrient conditions than did larvae from white masses (Figures (d, e, f) below).

Fig1C-f

The relationship between nutrient level and (c) days to hatching, (d) snout-vent length (the distance between the tip of the snout and the cloacal opening), (e) total length and (f) body mass.

These findings indicate that the polymorphism is advantageous in environments with considerable variation in nutrient levels. The white morph tends to do well at low nutrient levels, while the clear morph does better at higher nutrient levels. Pintar and Resetarits suggest that these differences in growth and development are likely to translate to higher adult survival and reproductive rates. The researchers used population modeling to demonstrate that under realistic conditions in which some individuals migrate from one pond to another, both morphs will persist indefinitely in ponds of varying nutrient levels.

We still don’t know why the two morphs perform differently under these different conditions. We do know that the outer jelly layer of white egg masses have white crystals made of proteins that are not water soluble, while the outer jelly layer of clear egg masses have smaller water soluble proteins. Pintar speculates that the firmer consistency of white egg masses could cause them to degrade more slowly and to retain their constituent nutrients more effectively than do the clear morphs.

note: the paper that describes this research is from the journal Ecology. The reference is Pintar, Matthew R., and William J. Resetarits Jr. “Persistence of an egg mass polymorphism in Ambystoma maculatum: differential performance under high and low nutrients.” Ecology (2017). The print version will probably come out in May or June of this year. Meanwhile, you can access it here. Thanks to the Ecological Society of America for allowing me to use figures from the paper. Copyright © 2017 by the Ecological Society of America. All rights reserved.

Cottonwood genes and spider hostels

Back in my working days, people that I met would sometimes ask me what I did for my research. I usually told them that I studied spider sex, which, while true, was a bit misleading, as my interests were actually slightly broader. But studying spider sex was a good fit for my disposition, because, more than anybody I know, I can stare at something for a very long time and not get bored. And when spiders have sex, there can be very long periods, when, to our eyes, nothing is going on. As it turns out there is a great deal of pheromonal communication going on, and considerable vibrational activity as well, but it is not that easy for humans (even abnormally patient ones) to detect.

Taphrina

A spider and her three egg cases within a web she has built in a Taphrina blister. Credit Matt Barbour

The point is that I have a very soft spot in my heart for spiders, and was delighted to learn about an indirect process that provided a comfortable home for needy spiders. Heather Slinn got interested in her project while an undergraduate summer intern. Her colleague, Matt Barbour, pointed out that when he flipped over blistered black cottonwood leaves (Populus trichocarpa) he often found a spider hanging out in there. This observation led to her to study the relationship between cottonwood trees, cottonwood genetics, pathogens that make leaf blisters and spider occupancy rates.

Taphrina fungi form cup-like blisters in the leaves of Populous trees. But these trees vary in how susceptible they are to leaf blisters. The researchers wanted to answer three questions about this relationship. First, do spiders prefer to live in leaf-blisters as opposed to unblistered leaves? Second, are differences in tree susceptibility to Taphrina a result of genetic differences between the trees? And third can differences in Taphrina-resistance account for differences in spider density?

One of the keys to the experiment was establishing a garden with distinct clones of trees of known genetic makeup (genotypes). Slinn and her colleagues studied five different genotypes, with approximately 40 trees per genotype. They dutifully watered them throughout the summer, and then sampled up to 30 leaves from each tree for blister density, blister size, and spider residency, using a modified shop vac to suck up all of the spiders.

DSCN4521

Black Cottonwood garden. Credit Matt Barbour

The researchers discovered that blistered leaves were 35 times more likely to have a spider and/or spider web than were unblistered leaves. Clearly spiders found blistered leaves to be highly attractive homes.

But there were pronounced differences among the five genotypes in blister density, blister size, spider density and the probability that a spider was occupying a blister. Graph A shows that genotypes 1 and 3 (G1 and G3) had the lowest mean density of blisters (about 2 or 3 per meter of plant), while G4 averaged about 20 blisters per meter. Although G3 had relatively few blisters, it did boast the largest blisters (see graph B). The researchers concluded that blister density and size were both under genetic control, but not linked to each other.

SlinnGraph1D

Mean (A) blister density (number per meter of plant) and (B) blister size, for the five tree genotypes.

But how did blister density and size influence spider residency? G4, the genotype with the greatest number of blisters per plant, also had the greatest number of spiders (graph C). But on a per blister basis, we can see that the two genotypes with the largest blisters (G2 and G3 – see graph B) also had the highest probability of housing a spider in their blisters (graph D). So when spiders make decisions about where to live, both size and number are important.

SlinnGraph2D

Mean (C) spider density and (D) probability a blister hosts a spider, for the five tree genotypes.

It is unclear why the spiders are attracted to blisters.  Some spider species have been shown to be attracted to structural complexity, because that provides more or better attachment points for web strands and egg cases.  Depressed blisters may also give protection from abiotic factors such as wind and precipitation.

Like most good studies, this research raises a host of related questions. Why is there so much genetic variation in Taphrina-resistance within this tree species? Slinn suggests that there may be tradeoff whereby investment into Taphrina-resistance might compromise a plant’s ability to invest in other functions such as cold-resistance, or rapid growth and/or high reproductive rates. A second question is how does spider presence influence other species – for example does hosting a spider reduce the number of herbivorous insects that might attack the tree? A third issue raised by the authors is that plants infected with Taphrina may be weaker and more susceptible to herbivores. In that case, the spiders may be present in blisters because they are attracted to the herbivores that are eating the leaves. And finally, herbivores eating the leaves may transmit other diseases affecting our forests, such as dutch elm disease, chestnut blight and white pine blister rust. Unraveling this complex chain of events will keep researchers busy for many years.

note: the paper that describes this research is from the journal Ecology. The reference is Slinn, H. L., Barbour, M. A., Crawford, K. M., Rodriguez‐Cabal, M. A., & Crutsinger, G. M. (2016). Genetic variation in resistance to leaf fungus indirectly affects spider density. Ecology 98(3): 875-881. Thanks to the Ecological Society of America for allowing me to use figures from the paper. Copyright © 2017 by the Ecological Society of America. All rights reserved.