Research Article |
Corresponding author: Adriana Sanchez ( adriana.sanchez@urosario.edu.co ) Academic editor: Jack Neff
© 2015 Adriana Sanchez, Edwin Bellota.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Sanchez A, Bellota E (2015) Protection against herbivory in the mutualism between Pseudomyrmex dendroicus (Formicidae) and Triplaris americana (Polygonaceae). Journal of Hymenoptera Research 46: 71-83. https://doi.org/10.3897/JHR.46.5518
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Herbivory significantly impacts the growth and reproduction of plants. Many plants have developed ways to defend against herbivores and one common strategy is to associate with ants. In many ant-plant interactions, ants are known to protect their host. However, in the Neotropical ant-plant genus Triplaris, the benefits provided by associated ants have never been tested. Many Pseudomyrmex spp. ants are obligate inhabitants of Triplaris spp. trees. In this study, Triplaris americana was studied in association with Pseudomyrmex dendroicus, an ant highly specific to its host (it has not been collected from any other species of Triplaris). Ant exclusion experiments were carried out to assess the protective effect of ants. In addition, ant behavior was monitored in control plants to study the mechanisms by which ants might confer protection against herbivory. Ant removal led to a more than 15-fold increase in herbivory. Pseudomyrmex dendroicus are active at all times of day and night and aggressively and efficiently remove insect herbivores from their host.
Ant-plant interaction, defense, herbivory, mutualism, myrmecophyte, Peru
Herbivory can have significant negative effects on plant fitness. Both the incidence and the impact of damage can vary with leaf ontogeny and the specific tissue being attacked, and herbivory can result in decreased survivorship and reproductive output (
Myrmecophytes (i.e., plants sheltering the colonies of a limited number of ‘plant-ant’ species in hollow structures called domatia and sometimes also providing them with food in the form of extrafloral nectar and food bodies) are pervasive and very diverse in the Tropics (
Several studies have shown that herbivory increases when ants are excluded from their host (e.g.,
Myrmecophytic-plant genera that have received much attention in the literature include Acacia, Cecropia, Macaranga, Duroia, and Cordia. However, several other genera remain to be studied. One such case is the myrmecophyte Triplaris. Some aspects of the ecological interaction between Triplaris and its associate ants have been addressed, but they have been limited to understanding the pruning behavior of Pseudomyrmex (
Questions regarding the effectiveness with which ants associated with Triplaris protect the plant against herbivores remain unexplored. The association between T. americana L. (Polygonaceae) and P. dendroicus Forel (Formicidae) is particular in that it displays high levels of specificity. After examining more than 200 collection records of Triplaris and its associated ants (A. Sanchez unpublished data), it was clear that P. dendroicus only colonizes individuals of T. americana, even when other species of Triplaris occur in sympatry. Therefore, in this study we expected a dramatic reduction in herbivory afforded by Pseudomyrmex ants compared to an ant-exclusion experiment and high and effective levels of protection.
This study was carried out at Los Amigos Biological Station (12°34'9"S, 70°6'0.40"W; ca 250 m) in the department of Madre de Dios in southeastern Peru. Los Amigos conservation area is a private conservation concession established in 2000 by the Peruvian government in conjunction with the Amazon Conservation Association (ACA) and the Asociación para la Conservación de la Cuenca Amazónica (ACCA). The station comprises more than 145.000 ha of lowland Amazonian forest between 250-320 m in altitude, at the confluence of the Madre de Dios and Los Amigos rivers, and protects pristine ecosystems including wetlands, seasonally inundated and terra firme forests, and palm swamps. The climate is characterized by a single dry and wet season each year. The area receives most of its estimated 2000 mm of annual rainfall during the wet season, which typically lasts from November to May (
The species chosen for this study were Triplaris americana and Pseudomyrmex dendroicus. Triplaris americana is the most common and widespread species in the genus. It is found from Panama to Bolivia and Brazil, usually in lowlands and disturbed areas close to water (
Prior to conducting the ant-exclusion experiments we explored two methods of exclusion, in order to determine which was the most effective. Following previous ant-exclusion experiments (e.g.,
To quantify the effects of ants on herbivory, two fully expanded leaves per sapling were monitored, always choosing the third leaf from the apical meristem from two adjacent branches. Prior to ant exclusion, photographs of every leaf were taken using a digital camera. A transparent sheet subdivided in grids of 1 square cm, each with 25 equidistant points within, was placed on top of the leaf, and photographs were always taken from the same distance. Percentage of herbivory was calculated by counting all the points that fell on areas where there was herbivory and divided by the total number of points that covered the leaf area. Photographs of each leaf were taken every two weeks for a total of six weeks.
We conducted a non-parametric Mann-Whitney U test for two independent samples to compare percentage of herbivory between control and experimental plants, using SigmaPlot version 12.5 (Systat Software Inc, San Jose, CA). We compared average herbivory after two, four and six weeks. The two leaves per sapling were used to calculate an average percentage of herbivory for each individual. A non-parametric ANOVA for repeated measures (Friedman ANOVA) was also conducted, to test if there was an increase of herbivory through time in the control and the ant-excluded plants.
Prior to taking the photographs, we conducted observations on the ants’ behavior, recording their patrolling activities and their interactions with potential herbivores and with other ants that occasionally visit Triplaris. Plants were monitored for approximately 5 to 10 min every visit. Since ants seem to have patrolling activities that span the 24-hour cycle, observations were also recorded for some plants before dusk (between 1600 h and 1700 h) and at night (between 2100 h and 2200 h).
Removal of ant colonies resulted in an increase in the percentage of herbivory. In each time interval (after 2, 4, and 6 weeks) there was a significant increase in herbivory compared to the control (U = 14, P < 0.05; U = 21; P < 0.05; U = 14.5, P < 0.01 respectively; Fig.
Seven out of eleven control plants had zero percent herbivory during the six weeks of the experiment. The four other control plants suffered some herbivory by weeks 4 and 6 (~ 1%). In contrast, of the ant-excluded plants, six plants had more than 3% herbivory, having as high as 16% leaf damage (outlier not shown; Fig.
Percentage of herbivory with time for the control and ant-excluded plants (grey). Removing the ant colonies resulted in a significant increase in the percentage of herbivory after 2, 4, and 6 weeks (U = 14, P < 0.05; U = 21; P < 0.05; U = 14.5, P < 0.01 respectively). There was also a significant increase in herbivory through time in the ant-excluded plants (ANOVA χ2N=11, df=2 = 15.8, P < 0.001), but not in the control (ANOVA χ2N=11, df=2 = 4, P > 0.05).
Pseudomyrmex dendroicus actively patrolled their hosts, at all times of day and night, even when the temperature was as low as 13 °C. Whenever the plant was disturbed, they efficiently recruited other workers, and were very aggressive against any intruder. During their patrolling activities they removed any debris found on top of the leaves. They repeated this cleaning process constantly, on all the leaves of their host. In all saplings studied, the leaves had no signs of mosses, fungi or lichens growing them, and no sign of accumulated debris.
From our observations, the most common herbivores were grasshoppers (unidentified; Orthoptera) and caterpillars of the lepidopteran genera Lophocampa (Arctiidae, subfamily Arctiinae) and Hylesia (Saturniidae). When an ant encountered a caterpillar, a worker approached and detected it with its antennae, and then recruited more workers (Fig.
This is the first study to report that Pseudomyrmex provides benefits to Triplaris by reducing herbivory. Our results indicated a significantly higher percentage of herbivory on the plants where ants were excluded (more than 15 times more; Fig.
According to the optimal defense theory (
It has also been suggested that the protective role of ants extends to protecting the host against pathogenic fungi (
Ants constitute a rapid and direct line of defense, which can mobilize where they are required (
The effects of herbivory may also extend beyond growth, ultimately affecting reproductive success and fitness of the host. Decreased energy spent on reparative growth could translate into increased energy allocation towards reproduction (
Ant-exclusion experiments revealed that the myrmecophyte Triplaris americana is significantly affected by herbivory in the absence of its symbiotic associate Pseudomyrmex dendroicus. Ants actively patrol their host at all times of day and night, and rapidly recruit when an herbivore is encountered. Even though caulinary domatia are costly to produce (
Funding for this study was provided by a grant from the Amazon Conservation Association. We are thankful to all the staff and people at Los Amigos, especially Nigel Pitman. We would also like to thank Megan Frederickson for her valuable comments and guidance through the experiment, and Antonio Coral and Brian Phillips for assistance in the field. Philip Ward kindly helped with identification of the ants, Lee Dyer and David Wagner with caterpillar identification. We would like to thank Doyle McKey and Alain Dejean for their helpful comments during the review process. INRENA-Ministerio de Agricultura in Peru provided the permits necessary for carrying out the experiments in Madre de Dios.