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.


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.

Saguaro survival: establishing an icon

Having grown up in the New York metropolitan area, my only contact with the saguaro cactus, Carnegiea gigantea, was from several TV westerns, which dubiously placed these mammoth cacti in New Mexico, Texas and Colorado.  In fact, the saguaro is limited to the Sonoran Desert of northwestern Mexico, extreme southeast California and southern and central Arizona. You won’t find these cacti further north, because a freeze lasting more than 24 hours kills them.  I still remember my first real sighting of these cacti; I was amazed at how distinct they seemed in comparison to the other vegetation, and I delighted in their abundance.

Daniel Winkler - Saguaro Photo 1

Dense patch of saguaros. Credit: Daniel Winkler

Many others delight in their abundance as well.  The flowers, fruits and seeds feed many animals (including humans).  They were an important food for the Tohono O’odham and Pima Indians – eaten fresh or converted into numerous products including wine, juice, jam and syrup.

Daniel Winkler - Saguaro Photo 2

Large saguaro with many fruits emanating from the apex of its branches. Credit: Daniel Winkler

Woodpeckers and flickers excavate nests in the saguaro’s trunk, which are subsequently occupied by other animals such as snakes, arthropods and small mammals.


Saguaro with nest cavity excavated near the top of its trunk. Credit: Daniel Winkler

Daniel Winkler also delighted in the saguaro’s awesomeness. As he describes “I fell in love with answering some basic ecology questions about the saguaro. I was surprised that scientists had been studying this wonderful plant for almost 100 years and there were still many basic questions about the species general biology and ecology that remained unanswered. Thus, I was hooked immediately and became obsessed with saguaro.”

Don Swann - Photo of D. Winkler with young saguaros

Daniel Winkler with young saguaros. Credit: Don Swann

Winkler and his colleagues wanted to know how moisture, temperature and habitat influence the establishment or survival of juvenile saguaro seedlings. Previous research had shown that saguaro height can be used to estimate saguaro age, given knowledge of previous rainfall in a particular area. So buoyed by an army of citizen scientists whom they recruited with the help of social media, student groups from schools and volunteers working at the Saguaro National Park, the research team estimated the age of every saguaro on 36 4-ha plots (1 ha = 10,000 m2).

Going into the study, the researchers knew that rainfall was a very important factor, with saguaros surviving better during wet periods.  But they also knew that historically, some areas located near each other showed different establishment trends, thus they suspected that other variables, particularly land use and other landscape factors, might be important.  They did their research in two different districts within the park: 21 plots in the Rincon Mountain District (RMD) on the east side of the park, and 15 plots in the Tucson Mountain District (TMD) to the west. They classified each plot as a particular habitat type based on slope, elevation and soil-type. Bajada was low elevation, flat and had gravelly porous soils.  Foothills were intermediate elevation and intermediate slope, while sloped habitats had highest elevation, steepest slope, and the coarsest rockiest soils.

Daniel Winkler - Saguaro Photo 4

Panoramic view of Saguaro National Park showing diversity of habitats. Credit: Daniel Winkler.

Winkler and his colleagues calculated the Palmer Drought Severity Index (PDSI) for the years 1950-2003. The PDSI quantifies the water balance between precipitation and evapotranspiration, taking into account not only rainfall but also other factors such as temperature and cloud cover.  The PDSI was estimated by assessing tree ring width for each year in nearby woodlands; wet conditions have wide tree rings (maximum PDSI value = +6), while dry years have narrow tree rings (minimum PDSI value = -6).

The researchers discovered a very strong association between the PDSI and seedling establishment. Low PDSI at the beginning and especially the end of the time frame was associated with low seedling establishment, while high PDSI (particularly in the 1980s was associated with high rates of seedling establishment (top graph below).  But other patterns emerged as well.  For example, establishment was higher in the TMD during the wettest years, but higher in the RMD during the most recent drought (bottom graph below).


Top. Total number of saguaros (left Y-axis) established per hectare from 1950-2003 in relation to PDSI (dashed line, right Y-axis). Bottom. Total number of saguaros established per hectare in the Tucson Mountain District (TMD – filled bars) and the Rincon Mountain District (RMD – open bars)  from 1950-2003 in relation to PDSI (dashed line, right Y-axis).

Saguaro establishment increased in all habitats when conditions were relatively wet (more positive PDSI values).  Under drought conditions, slopes had greatest saguaro establishment, while establishment increased more rapidly in foothills (and to a lesser extent in Bajadas) as moisture levels increased.


Model projecting number of saguaros established in the three major habitats in relation to PDSI.  Shaded regions are 95% confidence intervals.

The researchers were surprised at how tight the connection was between drought and saguaro establishment. But landscape features are also important.  The TMD is warmer and dryer than the nearby RMD, and had substantially lower establishment during the recent drought. The slopes in the RMD are steeper and rockier than sloped areas of the TMD, and may buffer saguaros from drought by capturing water in rock crevices and holding it for longer periods of time so it can be absorbed by saguaro roots. Nurse trees that provide shade to young saguaros may also be more common on the RMD slopes.

Winkler and his colleagues are concerned about the long-term impacts of climate change on saguaro populations, particularly in the drier areas of the TMD. They urge researchers to explore how long-term management of grazing and invasive species influences saguaro establishment across the landscape.  They also encourage researchers to gather some very basic data about saguaros, such as how they access water and how human water use patterns influence the water’s availability to this iconic species.

note: the paper that describes this research is from the journal Ecology. The reference is Winkler, D. E., Conver, J. L., Huxman, T. E. and Swann, D. E. (2018), The interaction of drought and habitat explain space–time patterns of establishment in saguaro (Carnegiea gigantea). Ecology 99: 621-631. doi:10.1002/ecy.2124. 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).


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.


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.


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.


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).


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.

Snails grow large to fight fear

In a recent post (Jan 12), I discussed research showing that song sparrow parents reduce provisioning to their offspring when threatened by predators, ultimately reducing offspring survival rates.  But in a turnabout that highlights the natural world’s dazzling diversity, a recent study by Sarah Donelan and Geoffrey Trussell revealed a very different impact of fear on the development of snail offspring. Donelan had worked as Trussell’s laboratory technician for two years and became fascinated by the egg capsules laid by the carnivorous snail Nucella lapillus, an ecologically important species in rocky intertidal communities. Earlier work had shown that predator-induced fear reduced snail feeding and growth rates, so Donelan decided that for her PhD work she would see how predator-induced fear influenced offspring development.


Adult Nucella alongside ca. 100 egg capsules. Credit: Sarah Donelan.

The researchers recognized that the fear environment experienced by parents before or during reproduction, and by the embryos during early development, could influence growth and development of those embryos. At their research site along the Massachusetts, USA coast, the predatory green crab, Carcinus maenas, can be a source of fear for these adult and embryonic snails. Donelan and Trussell exposed snails to fear by housing separately one male and one female snail in adjacent protected perforated containers (with six blue mussels in each container to feed them) that were set within a large plastic bucket. This bucket also had a somewhat larger perforated container (the risk chamber) containing the dreaded green crab (and two snails to feed it). The control risk chamber had two snails, but no crab.


Experimental setup with buckets containing egg capsules in perforated cages experiencing different exposure to fear. Credit: Sarah Donelan.

In late spring of 2015 and 2016, field-collected female and male snails were matched to create a total of 80 parental pairs. Donelan and Trussell set up experiments to explore the effects of parental experience with predation risk, embryonic experience with predation risk, and duration of embryonic experience.

Parent snails were exposed to a risk chamber (with a crab in the experimental group, and without a crab in the control group) for three days, and then placed together for four days (without risk) to mate. If an egg capsule was laid, the researchers removed it, and immediately exposed it to an experimental or control risk chamber for a week. Embryonic risk duration was further manipulated by continuing to expose half of the egg capsules to risk for a total of six weeks. The table below summarizes the treatments received by parents and offspring.



Mean (+ standard error) shell length (top graph) and tissue mass (bottom graph) of snail embryos exposed to predation risk. Parents were either exposed (solid circles) or not exposed (open circles) to risk before mating.


When parents were not exposed to risk, but their offspring were exposed, these offspring had shorter shells and reduced tissue mass compared to all other groups. When both parents and offspring were exposed to risk, offspring shell length increased by 8% and offspring mass increased by a whopping 40% over risk-exposed offspring whose parents were not exposed to risk (left data points in figures a and b). If embryos were not exposed to risk, parental exposure had no significant impact on embryonic development (right data points on figures a and b). Embryonic risk duration had no impact on development.


In addition, risk-exposed offspring of risk-exposed parents emerged from their egg capsules an average of 4.1 days sooner than other offspring.


Mean (+standard error) number of days until emergence of snail offspring that experienced the presence or absence of predation risk during early development.  Their parents were exposed to risk (solid circle) or no risk (open circle) before mating.

What could be causing these differences in size and rate of development? Donelan and Trussell hypothesized that embryonic snails could grow larger and more quickly if they were somehow able to reduce their metabolic rate. With a reduction in metabolic rate, more energy could be diverted to growth and development, resulting in larger and faster-growing snails. The researchers used an oxygen meter to measure oxygen consumption rates of individual egg capsules (from the eight different treatments in the first experiment) six weeks after deposition, about a week before embryos would begin to emerge. They exposed some of the capsules to predation risk during the experiment (current risk graph below), and left other capsules unexposed. When tested under risky conditions, capsules from parents who were exposed to risk, and that experienced risk as embryos during early development, had 56% lower metabolic rates than the other three groups (left graph), and similarly low metabolic rates as capsules tested without risk (right graph).


Mean (+ standard error) respiration rate of egg capsules that were (left graph) or were not (right graph) exposed to current predation risk.  During early development, the embryos in these capsules experienced risk or no risk, and were produced by parents exposed to risk (solid circles) or no risk (open circles) before mating.

Overall, parental experience with predation risk enhances offspring growth and development in the presence of risk. If the parents lack this exposure, risk-exposed offspring suffer the costs associated with small size and slower development. Currently Donelan and Trussell are trying to figure out what these costs are. Smaller snails have less energy reserves, may feed on a less diverse group of prey, and are less likely to remain in safer habitats than are larger juveniles. But we still don’t know whether these effects on early stages of life have lasting impacts as a snail gets older and larger. More generally, we don’t know whether there are similar types of interactions between parental and embryonic experiences of other stressors, most notably environmental stresses that are already being imposed by climate change.

note: the paper that describes this research is from the journal Ecology. The reference is Donelan, S. C. and Trussell, G. C. (2018), Synergistic effects of parental and embryonic exposure to predation risk on prey offspring size at emergence. Ecology, 99: 68–78. doi:10.1002/ecy.2067. 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.


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.


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.


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).


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.


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.


Homing in on the micro range

I’ve always been fascinated by geography. As a child, I memorized the heights of mountains, the populations of cities, and the areas encompassed by various states and countries. I can still recite from memory many of these numbers – at least based on the 1960 Rand McNally World Atlas. Part of my fondness for geography is no doubt based on my brain’s ability to recall numbers but very little else.

Most geographic ecologists are fond of numbers, exploring numerical questions such as how many organisms or species are there in a given area, or how large an area does a particular species occupy? They then look for factors that influence the distribution and abundance of species or groups of species. Given that biologists estimate there may be up to 100 million species, geographic ecologists have their work cut out for them.

As it turns out, most geographic ecologists have worked on plants, animals or fungi, while relatively few have worked on bacteria and archaeans (a very diverse group of microorganisms that is ancestral to eukaryotes).


Two petri plates with pigmented Actinobacteria. Credit: Mallory Choudoir.

Until recently, bacteria and archaeans were challenging subjects because they were so small and difficult to tell apart. But now, molecular/microbial biology techniques allow us to distinguish between closely related bacteria based on the sequence of bases (adenine, cytosine, guanine, and uracil) in their ribosomal RNA. Bacteria which are identical in more than 97% of their base sequence are described as being in the same phylotype, which is roughly analogous to being in the same species.

As a postdoctoral researcher working in Noah Fierer’s laboratory with several other researchers, Mallory Choudoir wanted to understand the geographic ecology of microorganisms. To do so, they and their collaborators collected dust samples from the trim above an exterior door at 1065 locations across the United States (USA).


Dr. Val McKenzie collects a dust sample from the top of a door sill. Credit: Dr. Noah Fierer.

The researchers sequenced the ribosomal RNA from each sample to determine the bacterial and archaeal diversity at each location. Overall they identified 74,134 gene sequence phyloypes in these samples – that took some work.

On average, each phylotype was found at 70 sites across the USA, but there was enormous variation. By mapping the phylotypes at each of the 1065 locations, the researchers were able to estimate the range size of each phylotyope. They discovered a highly skewed distribution of range sizes, with most phylotypes having relatively small ranges, while only a very few had large ranges (see the graph below). As it turns out, we observe this pattern when analyzing range sizes of plant and animal species as well.


Mean geographic range (Area of occupancy) for each phylotype in the study.  The y-axis (Density) indicates the probability that a given phylotype will occupy a range of a particular size (if you draw a straight line down from the peak to the x-axis, you will note that most phylotypes had an AOO of less than 3000 km2

Taxonomists use the term phylum (plural phyla) to indicate a broad grouping of similar organisms. Just to give you a feel for how broad a phylum is, humans and fish belong to the same phylum. Some microbial phyla had much larger geographic ranges than others. Interestingly, it was not always the case that the phylum with the greatest phylotype diversity had the largest range. For example, phylum Chrenarchaeota had the greatest median geographic range (see the graph below), but ranked only 19 (out of 50 phyla) in number of phylotypes (remember that a phylotype is kind of like a species in this study).


Box plots showing range size distribution for individual phyla. Middle black line within each box is the median value; box edges are the 25th and 75th percentile values (1st and 3rd quartiles).  Points are outlier phylotypes. Notice that the y-axis is logarithmic.

With this background, Choudoir and her colleagues were prepared to investigate whether there were any characteristics that might influence how large a range would be occupied by a particular phylotype. We could imagine, for example, that a phyloype able to withstand different types of environments would have a greater geographic range than a phylotype that was limited to living in thermal pools. Similarly, a phylotype that dispersed very effectively might have a greater geographic range than a poor disperser.

The researchers expected that aerobic microorganisms (that use oxygen for their metabolism) would have larger geographic ranges than nonaerobic microorganisms, which are actually poisoned by oxygen. The data below support this prediction quite nicely.


Geographic range size in relation to oxygen tolerance.  In this graph, and the graphs below, the points have been jittered to the right and left of their bar for ease of viewing (otherwise even more of the points would be on top of each other).

Some bacterial species form spores that protect them against unfavorable environmental conditions. The researchers expected that spore-forming bacteria would have larger geographic ranges than non-spore-forming bacteria.


Geographic range in relation to spore formation (left graph) and pigmentation (right graph).

Choudoir and her colleagues were surprised to discover exactly the opposite; the spore forming bacteria had, on average, slightly smaller geographic ranges. Choudoir and her colleagues also expected that phylotypes that are protected from harsh UV radiation by pigmentation would have larger geographic ranges than unpigmented phylotypes – this time the data confirmed their expectations.

The researchers identified several other factors associated with range size. For example, bacteria with more guanine and cytosine in their DNA or RNA tend to have larger geographic ranges. Some previous studies have shown that a higher proportion of guanine and cytosine is associated with greater thermal tolerance, which should translate to a greater geographic range. Choudoir and her colleagues also discovered that microorganisms with larger genomes (longer DNA or RNA sequences) also had larger ranges. They reason that larger genomes (thus more genes) should correspond to greater physiological versatility and the ability to survive variable environments.

This study opens up the door to further studies of microbial geographic ecology. Some patterns were expected, while others were surprising and beg for more research. Many of these microorganisms are important medically, ecologically or agriculturally, so there are very good reasons to figure out why they live where they do, and how they get from one place to another.

note: the paper that describes this research is from the journal Ecology. The reference is Choudoir, M. J., Barberán, A., Menninger, H. L., Dunn, R. R. and Fierer, N. (2018), Variation in range size and dispersal capabilities of microbial taxa. Ecology, 99: 322–334. doi:10.1002/ecy.2094. 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.

“Notes from Underground” – cicadas as living rain gauges

Given recent discussions between Donald Trump and Kim Jong-un about whose button is bigger, many of us with entomological leanings have revisited the question of what insects are most likely to dominate a post-nuclear world. Cicadas have a developmental life history that predisposes them to survival in the long term because some species in the eastern United States spend many subterranean years as juveniles (nymphs), feeding on the xylem sap within plants’ root systems. Magicicada nymphs live underground for 13 or 17 years, depending on the species, before digging out en masse, undergoing one final molt, and then going about the adult business of reproduction. This life history of spending many years underground followed by a mass emergence has not evolved to avoid nuclear holocausts while underground, but rather to synchronize emergence of billions of animals. Mass emergence causes predator satiation, an anti-predator adaptation in which predators are gastronomically overwhelmed by the number of prey items, so even if they eat only cicadas and nothing else, they still are able to consume only a small fraction of the cicada population.


Mass Magicicada emergence picturing recently-emerged winged adults, and the smaller lighter-colored exuviae (exoskeletons) that are shed during emergence. Credit: Arthur D. Guilani.

Less well-known are the protoperiodical cicadas (subfamily Tettigadinae) of the western United States that are abundant in some years, and may be entirely absent in others. Jeffrey Cole has studied cicada courtship songs for many years, and during his 2003 field season noted that localities that had previously been devoid of cicadas now (in 2003) hosted huge numbers of six or seven different species. He returned to those sites every year and high diversity and abundance reappeared in 2008 and 2014. This flexible periodicity contrasted with their eastern Magicicada cousins, and he wanted to know what stimulated mass emergence.



Protoperiodical cicadas studied by Chatfield-Taylor and Cole.  Okanagana cruentifera (top) and Clidophleps wrighti (bottom). Credit Jeffrey A. Cole.

Cole and his graduate student, Will Chatfield-Taylor, considered two hypotheses that might explain protoperiodicity in southern California (where they focused their efforts). The first hypothesis is that cicada emergence is triggered by heavy rains generated by El Niño Southern Oscillation (ENSO), a large-scale atmospheric system characterized by high sea temperature and low barometric pressure over the eastern Pacific Ocean. ENSO has a variable periodicity of 4.9 years, which roughly corresponds to the timing Cole observed while doing fieldwork. The second hypothesis recognized that nymphs must accumulate a set amount of xylem sap from their host plants to complete development. Sap availability depends on precipitation, and this accumulation takes several years in arid habitats. So while ENSO may hasten the process, the key to emergence is a threshold amount of precipitation over a several year timespan.

Working together, the researchers were able to identify seven protoperiodical species by downloading museum specimen data (including where and when each individual was collected) from two databases (iDigBio and SCAN). They also used data from several large museum collections, which gave them evidence of protoperiodical cicada emergences back to 1909. Based on these data, Chatfield-Taylor and Cole constructed a map of where these protoperiodical cicadas emerge.


Maps of five emergence localities discussed in this study.

The researchers tested the hypothesis that protoperiodical cicada emergences follow heavy rains triggered by ENSO by going through their dataset to see if there was a correlation between ENSO years and mass cicada emergences. Of 20 mass cicada emergences since 1918, only five coincided with ENSO events, which is approximately what would be expected with a random association between mass emergences and ENSO. Scratch hypothesis 1.

Let’s look at the second hypothesis. The researchers needed reliable precipitation data between years for which they had good evidence that there were mass emergences of their seven species. Using a statistical model, they discovered that 1181 mm was a threshold for mass emergences, and that three years was the minimum emergence interval regardless of precipitation. Only after 1181 mm of rain fell since the last mass emergence, summed over at least three years, would a new mass emergence be triggered.


Cumulative precipitation over seven time periods preceding cicada emergence.

The nice feature of this model is that it makes predictions about the future. For example, the last emergence occurred in the Devil’s punchbowl vicinity in 2014. Since then that area has averaged 182.2 mm of precipitation per year. If those drought conditions continue, the next mass emergence will occur in 2021 at that locality, which is longer than its historical average. Only time will tell. Hopefully Mr. Trump and Mr. Jong-un will be able to keep their fingers off of their respective buttons until then.

note: the paper that describes this research is from the journal Ecology. The reference is Chatfield-Taylor, W. and Cole, J. A. (2017), Living rain gauges: cumulative precipitation explains the emergence schedules of California protoperiodical cicadas. Ecology, 98: 2521–2527. doi:10.1002/ecy.1980. 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.