Open Access
How to translate text using browser tools
1 December 2014 Ants (Hymenoptera: Formicidae) and Trophobiont Leafhopper Nymphs (Hemiptera: Cicadellidae) Become more Abundant in Shaded Conditions: Implications for Mutualism
Gustavo Moya-Raygoza, Adelina Valle Martinez
Author Affiliations +

Abiotic factors have dramatic effects on herbivore insect populations. However, little is known about the effects of the abiotic factor of shade on the mutualism between ants and Hemiptera (aphids, scales, mealy bugs, whiteflies, treehoppers, leafhoppers). The trophobiont leafhopper Dalbulus quinquenotatus DeLong & Nault (Hemiptera: Cicadellidae) is obligatorily tended by the ant Brachymyrmex obscurior Forel (Hymenoptera: Formicidae) on the basal leaves of gamagrass Tripsacum dactyloides L. (Poales: Poaceae). To better understand the effect of abiotic factors on this relationship, we conducted a comparative experiment to investigate the abundance of ants and leafhoppers (nymphs and adults) under shaded and full-sunlight conditions. We observed mutualism between D. quinquenotatus and B. obscurior on both shaded and non-shaded gamagrasses. Nevertheless, the greatest number of B. obscurior were found tending the greatest number of D. quinquenotatus on shaded T. dactyloides. Most of these leafhoppers were nymphs. Nymphs were observed in clusters on the basal leaves. On the other hand, ant-tended D. quinquenotatus adults were observed at similar frequencies on shaded and non-shaded gamagrasses. The adults were also seen on the basal leaves and produced honeydew, but were more mobile than the nymphs. Our results highlight the importance of abiotic factors in mutualism as well as the complex interaction between shaded plants, immature leafhoppers, and tending ants.

Most ants depend on sugar secretions as a food resource, and these secretions are produced in great quantity in shaded habitats (Perfecto et al. 1996; Vandermeer et al. 2002; de la Mora et al. 2013) by herbivorous insects, including trophobiont hemipterans (Moya-Raygoza & Larsen 2008). Honeydew is a sugary excretion of carbohydrates, amino acids, and water that attracts ants to establish a mutualistic association. The hemipterans offer food in the form of honeydew, and the ants provide protection to the hemipterans against predators and parasitoids (Way 1963; Buckley 1987; Blüthgen et al. 2006; Gibb & Cunningham 2009; Zhang et al. 1012; Zhang et al. 2013). Ants are abundant when the population of hemipterans is large because of the increased quantity of honeydew (Katayama & Suzuki 2010). With increased production of honeydew resulting in increased visits by tending ants (Bronstein 1994; Itiokia & Inoue 1996), honeydew is the base on which ant-hemipteran mutualism is built (Wiss 2006; Styrsky & Eubanks 2007; Detrain et al. 2010).

To our knowledge, no study to date has investigated whether abundance of ants is associated with abundance of immature (nymph) or mature (adult) stages of trophobiont leafhoppers on shaded host plants. We hypothesized that shade would affect the intensity of mutualism by increasing the number of leafhopper nymphs and tending ants on gamagrass Tripsacum dactyloides L. (Poales: Poaceae). Immature leafhoppers are the keystone in the association with ants in shaded habitats, because of the great food resources offered to ants. Wood (1982) found nymphs of the treehopper Enchenopa binotata Say (Membracidae) in great aggregation with abundant tending ants, and Cushman & Whitham (1989) present evidence that large aggregations of nymphs benefit more from ant tending than small aggregations. Neither study, however, investigated the effect of shade. In the present study, we focus in the hemipteran trophobiont leafhopper Dalbulus quinquenotatus DeLong & Nault (Hemiptera: Cicadellidae), which occurs in obligatory mutualism with the ant Brachymyrmex obscurior Forel (Hymenoptera: Formicidae). This leafhopper-ant mutualism takes place on the basal leaves of the gamagrass host plant (Tripsacum spp.) (Nault et al. 1983; Larsen et al. 1991; Moya-Raygoza & Nault 2000) in shaded and sunny habitats. Dalbulus quinquenotatus is sedentary and gregarious, and produces large quantities of honeydew. This honeydew is a food resource for ants, which in return protect D. quinquenotatus from natural enemies (Moya-Raygoza & Nault 2000; Larsen et al. 2001). The objective in this study was to examine the abundance of ants and leafhoppers (nymphs and adults) on shaded and unshaded T. dactyloides plants.

Materials and Methods

This experiment, comparing the effects of shade and non-shade conditions, was conducted in Zapopan, state of Jalisco, Mexico (N 20° 44′ -W 103′) at 1,570 m elevation. Gamagrass (T. dactyloides) plants were obtained by rhizome propagation. Rhizomes were collected from natural Zapopan gamagrass populations and grown in pots (30 cm in diam × 33 cm height). The pots contained the same type of soil as that in which the gamagrass grows naturally in the field. We used 30 pots, divided into 2 treatments: 15 pots for the shade treatment and 15 pots for the nonshade (exposed to full sunlight) treatment. Each pot contained green young plants that had recently emerged from the rhizomes. The plants were the same age and had either 2 or 3 stems. Each pot was considered one replicate. The shade treatment group included a total of 34 stems and the non-shade treatment group included a total of 32 stems. The pots with the plants were arranged in 2 circles, one for each treatment. For the shade treatment, a mesh was installed that produced 45% shade. The mesh was 3 m high and covered the 15 pots in the shaded treatment; no shade was installed in the non-shaded treatment. The distance between treatments was 20 m (Fig. 1). First we installed the mesh, and then the 30 pots divided into 2 treatments; 3 days later D. quinquenotatus adults were deposited on the T. dactyloides plants.

Dalbulus quinquenotatus adults were collected with an aspirator from a T. dactyloides field population located at Zapopan. The collected adults were gender separated in the lab and deposited on the basal leaves of the gamagrass plants reared in the pots. Twenty adults (10 females and 10 males) of D. quinquenotatus were deposited in each pot. Each pot was covered with a screen for 2 weeks to allow adults to oviposit and become established on the basal leaves. After the 2 weeks, the screens were removed and one additional pot per treatment, containing a nest of B. obscurior of similar size, was installed in the center of each circle. Similar abundance of ants between the 2 nests was evident when ants were connected to the leafhoppers as shown in Fig. 5. The distance from the nest to each pot was 1 m, so that the ants had to traverse the same distance to reach leafhoppers in all treatment. After 2 weeks, nymphs were present. Dalbulus quinquenotatus development time from egg to adult is about 34 days at 24 °C. Nymphs begin hatching on day 12 and nymphal abundance peaks on day 16 (Moya-Raygoza 2004). No arthropod predators or parasitoids such spiders, hemipterans or dryinids (Hymenoptera: Dryinidae) that may attack D. quinquenotatus were observed during the experiment. Also, no plant with extrafloral nectaries that may attract ants was present in the experimental area.

After 2 weeks, when the nymphs appeared, counting of nymphs, adults, and ants on each stem from the 2 treatments commenced. The counting of all the insects was conducted without disturbing them, between 10:00 and 13:00 hours, every 7 days. To avoid variation the counting was conducted randomly on each collection date. The time in observing each plant was between 2 to several minutes and it was facilitated because mutualism was given only on the basal leaves of the gamagrass on a leaf segment no longer than 5 cm. The experiment was conducted between 11 Oct and 29 Nov 2006. The mean daily high temperature was 25.56 °C (SE = 0.31). Plants were watered every 4 days. Repeated-measures analysis of variance (ANOVA) on transformed (χ2) numbers of ants and nymph and adult leafhoppers was used to compare these quantities between the 2 treatments over time. All statistical analysis was performed using SPSS software (SPSS 11.5 for Windows, SPSS, Chicago, IL).

Fig. 1.

Experimental design. In each treatment, 15 pots (black circles) were used, each pot containing the host plant Tripsacum dactyloides and 10 males and 10 females Dalbulus quinquenotatus. The central white circle represents the ant nest, the source of the tending ants, Brachymyrmex obscurior. After initiation of the experiment, the ants were free to move to the surrounding pots containing T. dactyloides and D. quinquenotatus.



The leafhopper D. quinquenotatus and the ant B. obscurior occurred in both shaded and unshaded treatments. However, significantly more leafhoppers were tended by B. obscurior on the shaded T. dactyloides over the course of 49 days (repeated-measures ANOVA; F = 3.00; df = 1,7; P = 0.004) (Fig. 2).

The difference in the numbers of leafhoppers tended appeared to be related to the abundance of nymphs. We found significantly more nymphs, tended by B. obscurior, in the shaded treatment than in the unshaded treatment (repeated-measures ANOVA; F = 2.73; df = 1,7; P = 0.01) (Fig. 3). Nymphs emerged after placement of adults in the covered pots during the first 2 weeks; before introducing the ants. These nymphs had soft bodies and produced honeydew after the first stadium. The number of ant-tended D. quinquenotatus adults, however, was similar in shaded and nonshaded treatments (repeated-measures ANOVA; F = 0.81; df = 1,7; P = 0.58) (Fig. 4). These adults produced honeydew and were more mobile than the nymphs. In both treatments, more adults were found between days 35 and 49, after all of the nymphs had completed their development.

The numbers of ants also differed between the 2 treatments. Significantly more ants were found tending D. quinquenotatus on the basal leaves of shaded T. dactyloides plants, throughout the 49 days (repeated-measures ANOVA; F = 2.67; df = 1,7; P = 0.01) (Fig. 5). The largest number of ants and nymphs in the shaded treatment occurred during the first 35 days, before all the nymphs became adults (Figs. 3 and 5).


We found significantly larger numbers of the leafhopper D. quinquenotatus on the shaded T. dactyloides plants, tended by significantly greater numbers of B. obscurior, compared with T. dactyloides plants without shade. Similar results were reported by Moya-Raygoza & Larsen (2008), who found large numbers of D. quinquenotatus and B. obscurior on gamagrass plants exposed to 50% shade in artificially constructed habitats. In addition, this positive effect of shade in the mutualism between ants and D. quinquenotatus was observed in nature, i.e., increased numbers of ants and leafhoppers were found in shelters built by ants in wild gamagrass (Moya-Raygoza & Larsen 2008). Grasses under shade have advantages in terms of water and nitrogen that in turn support herbivorous insects. For example, shaded conditions result in stomatal closure, reduced transpiration, and decreased water loss in T. dactyloides (Fay & Knapp 1995). In grasses, such as Axonopus compressus (Sw.) P. Beauv and Pennisetum clandestinum Hochst. ex Chiov., shade increases their nutritive value for herbivores through an increase in nitrogen content (Samarakoon et al. 1990). Therefore, shaded grasses are superior food resources for phloem-feeding hemipterans such as leafhoppers, because they depend entirely on a liquid diet and nitrogen from the host plant for survival and reproduction.

Fig. 2.

Mean (± SE) number of total Dalbulus quinquenotatus leafhoppers per stem on unshaded and shaded Tripsacum dactyloides. Day 1 is when nymphs first appeared.


Many trophobiont hemipterans produce copious excretions of honeydew to attract tending ants. Positive density-dependent ant-hemipteran mutualisms, such as we observed, have been associated with honeydew production of several trophobiont hemipterans. Cushman & Whitham (1989) observed that the population of the ant Formica altipetens Wheeler increased along with increased abundance of the membracid Publilia modesta Uhler, because large aggregations of the hemipteran, produce more honeydew and thus attracted more ants. Katayama & Suzuki (2003) found that the number of Tetramorium caespitum L. and Lasius niger L. ants increased as the density of the aphid Aphis craccivora Koch (Aphididae) increased, mainly because of copious honeydew production. Similarly, Fischer et al. (2001) found that the aphid Metopeurum fuscoviride Stroyan is preferred by L. niger ants for its abundant production of honeydew. Finally, Itioka & Inoue (1996) reported that the abundance of L. niger increases with the abundance of the scale insect Ceroplastes rubens Maskell (Coccidae).

Positive density-dependence has been reported in hemipteran nymphs and their tending ants. Wood (1982) found large aggregations of the treehopper nymphs E. binotata with large aggregations of tending ants. Cushman & Whitham (1989) provide evidence that large nymphal aggregations benefit more from ant tending than small nymphal aggregations, and Fischer et al. (2002) found that the abundance of L. niger ants is correlated with the amount of honeydew produced by third and fourth instar M. fuscoviride nymphs.

As mentioned before, to our knowledge no study has investigated the role of shade in the abundance of immature trophobiont hemipterans and their tending ants. We found that large numbers of leafhopper nymphs were associated with large numbers of ants on shaded T. dactyloides, when compared with plants exposed to full sunlight. The greater abundance of nymphs might be related in part to the increased water and nitrogen content of T. dactyloides in the shaded treatment. This would be in accordance with the findings of Ross et al. (1991) that the leafhopper Cameocephala floridana Ball increases in abundance and body size when feeding on shaded grasses with high nitrogen content. Future studies are needed to demonstrate the effect of water and nitrogen on the abundance in nymphs of D. quinquenotatus.

Fig. 3.

Mean (± SE) number of Dalbulus quinquenotatus leafhopper nymphs per stem on unshaded and shaded Tripsacum dactyloides. Day 1 is when nymphs first appeared.


Fig. 4.

Mean (± SE) number of Dalbulus quinquenotatus leafhopper adults per stem on unshaded and shaded Tripsacum dactyloides. Day 1 is when nymphs first appeared.


Fig. 5.

Mean (± SE) number of Brachymyrmex obscurior ants per stem tending Dalbulus quinquenotatus leafhoppers on unshaded and shaded Tripsacum dactyloides. Day 1 is when nymphs first appeared.


After D. quinquenotatus eggs hatch, nymphs appear and start to feed on the basal leaves of T. dactyloides. In shaded gamagrass, nymphs cluster on the leaves, and the removal of honeydew by ants reduces the death of nymphs by suffocation from accumulated honeydew and the formation of sooty mold. Young trophobiont hemipterans can become asphyxiated by the sticky liquid and molds if ants are absent or sparse (Wiss 2006). Moreover, nymphs in large aggregations with consequently large populations of ants are better protected in shaded gamagrass. These nymphs have soft bodies susceptible to predation if they are not protected by ants. In natural conditions, D. quinquenotatus nymphs live in shelters made by Solenopsis geminata (F.) (Formicidae) in shaded habitats that protect the nymphs from dryinid wasp parasitoids (Moya-Raygoza & Larsen 2008). Cushman & Whitham (1989) investigated the tending ant Formica altipetens Wheeler and the membracid P. modesta, and found that only nymphs, and not adults, benefit from ant tending, because nymphs are susceptible to predation by spiders, whereas adults are more agile and capable of escaping from predation. Nymphs of D. quinquenotatus have little mobility. Therefore, they are better protected when ants are abundant.

The great abundance of leafhopper nymphs in the shaded habitat is the driving force that determines ant abundance and, in turn, mutualism. The hemipterans that produce honeydew are subject to selection pressure because they produce the resources that attract ants (Yao et al. 2000). Styrsky & Eubanks (2007) and Detrain et al. (2010) concluded that hemipteran trophobionts are the keystone on which mutualism is built. In our case, an abiotic factor, shade, enhances that mutualism. Wiss (2006) argues that hemipteran trophobiont populations, with their honeydew that is rich in sugars and amino acids, are targets for selection and this affects the ecological interactions of these organisms. A large number of trophobiont hemipterans dramatically change the structure of the arthropod community because tending ants alter the abundance and distribution of predators, parasitoids, and herbivorous insects (Styrsky & Eubanks 2007).

The effect of abiotic factors on herbivore insect populations has been studied for many years (Masters et al. 1998). However, this is one of the first studies to evaluate the effect of abiotic factors on mutualism between hemipteran trophobionts and ants. Abiotic factors affecting mutualism between ants and plants bearing extrafloral nectaries, such as temperature and precipitation, have received little attention in spite of the dramatic effects such factors can have on that mutualism (Rico-Gray et al. 2012). Extrafloral nectaries and trophobiont hemipterans are analogous food resources because both produce secretions specifically developed to be utilized by ants. Future studies examining the role of abiotic factors affecting these 2 food resources consumed by ants, which generally co-occur in the same community, would help to further characterize these mutualistic relationships.


We thank Claudia S. Copeland (Carpe Diem Biomedical Writing and Editing) for editing the manuscript and Victor Rico-Gray for the comments and suggestions to the manuscript. Temperature information was provided by the Observatorio Meteorológico (Universidad de Guadalajara).

References Cited


N. Blüthgen , D. Mezger , and K. E. Linsenmair 2006. Ant-hemiptera trophobiosis in a Bornean rainforest-diversity, specificity and monopolization. Insect. Soc. 53: 194–203. Google Scholar


R. C. Buckley 1987. Interactions involving plants, Homotera, and ants. Annu. Rev. Ecol. Syst. 18: 111–135. Google Scholar


J. L. Bronstein 1994. Our current understanding of mutualism. Q. Rev. Biol. 69: 31–51. Google Scholar


J. H. Cushman , and T. G. Witham 1989. Conditional mutualism in a membracid-ant association: temporal, age-specific, and diversity-dependent effects. Ecology 70: 1040–1047. Google Scholar


A. De La Mora , C. J. Murnen , and S. M. Phipott 2013. Local and landcape drivers of biodiversity of four groups of ants in coffee landscapes. Biodivers. Conserv. 22: 871–888. Google Scholar


C. Detrain , F.J. Verheggen , L. Diez , B. Wathelet , and E. Haubruge 2010. Aphid-ant mutualism: how honeydew sugars influence the behavior of ant scouts. Physiol. Entomol. 35: 168–174. Google Scholar


P. A. Fay , and A. K. Knapp 1995. Stomatal and photosynthetic responses to shade in sorghum, soybean and eastern gamagrass. Physiol. Plantarum 94: 613– 620. Google Scholar


M. K. Fischer , K. H. Hoffmann , and W. Völkl 2001. Competition for mutualists in an ant- homopteran interaction mediated by hierarchies of ant-attendance. Oikos 92: 531–541. Google Scholar


M. K. Fischer , W. Völkl , R. Schopf , and K. H. Hoffmann 2002. Age-specific patterns in honeydew production and honeydew composition in the aphid Metopeurum fuscoviride: Implications for ant-attendance. J. Insect. Physiol. 48: 319–326. Google Scholar


H. Gibb , and S. A. Cunninham 2009. Does the availability of arboreal honeydew determine the prevalence of ecologically dominant ants in restored habitats?. Insect. Soc. 56: 405–412. Google Scholar


T. Itiokia , and T. Inoue 1996. Density-dependent ant attendance and its effects on the parasitism of a honeydew-producing scale insect, Ceroplastes rubens. Oecologia 106: 448–454. Google Scholar


N. Katayama , and N. Suzuki 2003. Changes in the use of extrafloral nectaries of Vicia faba (Leguminosae) and honeydew of aphids by ants with increasing aphid density. Ann. Entomol. Soc. America 96: 579–584. Google Scholar


N. Katayama , and N. Suzuki 2010. Extrafloral nectaries indirectly protect small aphid colonies via ant-mediated interactions. Appl. Entomol. Zool. 45: 505–511. Google Scholar


K. J. Larsen , F. E. Vega , G. Moya-Raygoza , and L. R. Nault 1991. Ants (Hymenoptera: Formicidae) associated with the leafhopper Dalbulus quinquenotatus (Homoptera: Cicadellidae) on gamagrasses in Mexico. Ann. Entomol. Soc. America 84: 498–501. Google Scholar


K. J. Larsen , L. M. Staehle , and E. J. Dotseth 2001. Tending ants (Hymenoptera: Formicidae) regulate Dalbulus quinquenotatus (Homoptera: Cicadellidae) population dynamics. Environ. Entomol. 30: 757–762. Google Scholar


G. J. Masters , V. K. Brown , I. P. Clarke , J. B. Whittaker , and J. A. Hollier 1998. Direct and indirect effects of climate change on insect herbivores: Auchenorrhyncha (Homoptera). Ecol. Entomol. 23: 45–52. Google Scholar


G. Moya-Raygoza 2004. Gamagrass leafhopper Dalbulus quinquenotatus Delong & Nault (Hemiptera: Cicadellidae), pp. 947–949 In J. L. Capinera (ed.), Encyclopedia of Entomology. Kluwer Academic Publishers. Google Scholar


G. Moya-Raygoza , and L. R. Nault 2000. Obligatory mutualism between Dalbulus quinquenotatus (Homoptera: Cicadellidae) and attendant ants. Ann. Entomol. Soc. America 93: 929–940. Google Scholar


G. Moya-Raygoza , and K. J. Larsen 2008. Positive effects of shade and shelter construction by ants on leafhopper-ant mutualism. Environ. Entomol. 37: 1471–1476. Google Scholar


L. R. Nault , D. M. Delong , B. W. Triplehorn , W. E. Styer , and J. F. Doebly 1983. More on the association of Dalbulus (Homoptera: Cicadellidae) with Mexican Tripsacum (Poaceae), including the description of two new species of leafhoppers. Ann. Entomol. Soc. America 76: 305–309. Google Scholar


I. Perfecto , R. A Rice, R. Greenberg , and M. E. Van Der Voort 1996. Shade coffee: A disappearing refuge for biodiversity. BioScience 46: 598–608. Google Scholar


V. Rico-Gray , C. Diaz-Castelazo , A. Ramirez-Hernandez , P. R. Guimaraes Jr. , and J. N. Holland 2012. Abiotic factors shape temporal variation in the structure of an ant-plant network. Arthropod Plant Interact. 6: 289–295. Google Scholar


A. M. Ross , and D. R. Strong 1991. Effect of hostplant nitrogen on preference and performance of laboratory populations of Carneocephala floridana (Homoptera: Cicadellidae). Environ Entomol. 20: 1349–1355. Google Scholar


S. P. Samarakoon , J. R. Wilson , and H. M. Shelton 1990. Growth, morphology and nutritive quality of shaded Stenotaphrum secundatum, Axonopus compressus and Pennisetum clandestium. J. Agric. Sci. 114: 161–169. Google Scholar


J. D. Styrsky , and M. D. Eubanks 2007. Ecological consequences of interactions between ants and honeydew-producing insects. Proc. R. Soc. B. 274: 151–164. Google Scholar


J. Vandermeer , I. Perfecto , G. Ibarra Nuñez , S. Philpott , and J. A. Garcia Ballinas 2002. Ants (Azteca sp.) as potential biological control agents in shade coffee production in Southern Chiapas, Mexico: Complication of insect effects. Agroforest. Syst. 56: 271–276. Google Scholar


M. J. Way 1963. Mutualism between ants and honeydew- producing Homoptera. Ann. Rev. Entomol. 8: 307–344. Google Scholar


M. R. Weiss 2006. Defecation behavior and ecology of insects. Annu. Rev. Entomol. 51: 635–661. Google Scholar


T. K. Wood 1982. Ant-attended nymphal aggregations in the Enchenopa binotata complex (Homoptera: Membracidae). Ann. Entomol. Soc. America 75: 649–653. Google Scholar


I. Yao , H. Shibao , and A. Kimoto 2000. Costs and benefits of ant attendance to the drepanosiphict aphid Tuberculatus quercicola. Oikos 89: 3–10. Google Scholar


S. Zhang , Y. Zhang , and K. Ma 2012. Distribution of ant-aphid mutualism in canopy enhances the abundance of beetles on the forest floor. PLoS ONE 7(4): e35468. Google Scholar


S. Zhang , Y. Zhang , and K. Ma 2013. The ecological effects of ant-aphid mutualism on plants at a large spatial scale. Sociobiology 60: 236–241. Google Scholar
Gustavo Moya-Raygoza and Adelina Valle Martinez "Ants (Hymenoptera: Formicidae) and Trophobiont Leafhopper Nymphs (Hemiptera: Cicadellidae) Become more Abundant in Shaded Conditions: Implications for Mutualism," Florida Entomologist 97(4), 1378-1385, (1 December 2014).
Published: 1 December 2014
Dalbulus, hormigas
hemípteros inmaduros
immature Hemiptera
tending ants
Back to Top