vol. 191, no. 4 the american naturalist april 2018 Natural History Note Venus Flytrap Rarely Traps Its Pollinators Elsa Youngsteadt,1,* Rebecca E. Irwin,2 Alison Fowler,2 Matthew A. Bertone,1 Sara June Giacomini,2 Michael Kunz,3 Dale Suiter,4 and Clyde E. Sorenson1 1. Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, North Carolina 27695; 2. Department of Applied Ecology, North Carolina State University, Raleigh, North Carolina 27695; 3. North Carolina Botanical Garden, University of North Carolina, Chapel Hill, North Carolina 27599; 4. US Fish and Wildlife Service, Raleigh, North Carolina 27606 Submitted August 14, 2017; Accepted October 10, 2017; Electronically published February 5, 2018 Online enhancements: appendix table. Dryad data: http://dx.doi.org/10.5061/dryad.p8s64. fl abstract: Because carnivorous plants rely on arthropods as polli- pollinator-prey con ict (Juniper et al. 1989; Zamora 1999; nators and prey, they risk consuming would-be mutualists. We exam- Jürgens et al. 2012). For example, recent experiments suggest ined this potential conflict in the Venus flytrap (Dionaea muscipula), that the red color of sundew (Drosera) traps reduces pollina- whose pollinators were previously unknown. Diverse arthropods from tor bycatch but at the expense of total prey capture (Jürgens two classes and nine orders visited flowers; 56% of visitors carried et al. 2015). With few exceptions, however, the extent and D. muscipula pollen, often mixed with pollen of coflowering species. strength of pollinator-prey conflict is poorly quantified (but Within this diverse, generalized community, certain bee and beetle see Zamora 1999; Murza et al. 2006; El-Sayed et al. 2016). species appear to be the most important pollinators, on the basis of their abundance, pollen load size, and pollen fidelity. Dionaea musci- The pollination systems of most carnivorous species are pula prey spanned four invertebrate classes and 11 orders; spiders, poorly documented, while the composition of their prey is of- beetles, and ants were most common. At the family and species levels, ten reported at coarse taxonomic resolution (Jürgens et al. few taxa were shared between traps and flowers, yielding a near-zero 2012). value of niche overlap for these potentially competing structures. Spa- Remarkably, Venus flytrap, Dionaea muscipula J. Ellis tial separation of traps and flowers may contribute to partitioning the (Droseraceae), is among the species whose pollinators have invertebrate community between nutritional and reproductive func- never been reported. Of more than 600 species of carnivo- tions in D. muscipula. rous plants in the world, D. muscipula is the only terrestrial Keywords: Venus flytrap, Dionaea muscipula, pollination ecology, species with an active snap trap (Ellison and Gotelli 2009). pollinator-prey conflict, niche overlap. Since D. muscipula came to the attention of Victorian nat- uralists in the 1800s, the rapid movement of its traps has prompted cultural fascination and extensive physiological Introduction study (e.g., Darwin 1875; Forterre et al. 2005; Chase et al. Plants typically interact with multiple species of animals 2009). The species is now cultivated in many countries for and exhibit ecological and evolutionary responses to the con- research, education, and horticultural trade, but its native flicting pressures imposed by mutualists (such as pollina- range is restricted to a small area of North Carolina and tors) and antagonists (such as herbivores; e.g., Strauss and South Carolina, where it is threatened by habitat loss, fire Irwin 2004; Sun et al. 2016). Carnivorous plants present a suppression, and poaching. The International Union for special case of potentially conflicting pressures. Because many Conservation of Nature and the North Carolina Plant Con- carnivorous plants rely on arthropods as pollinators and prey, servation Program list the species as vulnerable, and it has they risk consuming their would-be mutualists (Givnish 1989; been petitioned for listing under the US Endangered Species Ellison and Gotelli 2001). They may therefore face trade-offs Act (Schnell et al. 2000; Robinson and Finnegan 2016; between pollination services and nutrient intake, known as Waller et al. 2016). Despite the conservation status and unique trap biology of D. muscipula, its pollination ecology is relatively un- * Corresponding author; e-mail: [email protected]. known. From mid-May to early July, it produces white, ORCIDs: Youngsteadt, http://orcid.org/0000-0003-2032-9674; Fowler, http:// saucer-shaped flowers, approximately 2 cm in diameter, in orcid.org/0000-0002-9263-2253; Suiter, http://orcid.org/0000-0002-7494-7227; – Sorenson, http://orcid.org/0000-0002-7127-7272. a cymous raceme atop a scape 15 35 cm long (Smith 1929; Roberts and Oosting 1958). Roberts and Oosting (1958, Am. Nat. 2018. Vol. 191, pp. 000–000. q 2018 by The University of Chicago. 0003-0147/2018/19104-57880$15.00. All rights reserved. p. 202) reported that it is self-infertile and that pollination DOI: 10.1086/696124 is “presumably . entomophilous, apparently by various This content downloaded from 152.007.224.009 on February 05, 2018 12:03:04 PM All use subject to University of Chicago Press Terms and Conditions (http://www.journals.uchicago.edu/t-and-c). 000 The American Naturalist beetles, small flies, and possibly spiders,” although they did did not damage plants. We transferred prey into individual not present data or more specific taxonomic determina- vials of 70% ethanol. tions. Prey have been better quantified; spiders, ants, and We identified arthropods from flowers and traps to the beetles compose the largest share (Ellison and Gotelli 2009; lowest taxonomic level possible; target resolution was fam- Hutchens and Luken 2015). ily for spiders and Orthoptera (most of which were imma- The goal of this study was to document the potentially ture or poorly preserved in traps) and species/morphospe- conflicting insect interactions of D. muscipula flowers and cies for other insects. We used standard references (e.g., traps to inform further study of its evolutionary ecology Stehr 1987; Arnett et al. 2002 and references therein; Gibbs and conservation management. Specifically, we asked the 2011) and comparison to reference material in the North following: (1) Which invertebrate species visit D. muscipula Carolina State University Insect Museum, where voucher flowers? (2) Which of these are the most important poten- specimens will be deposited. tial pollinators? (3) To what extent do flowers and traps in- teract with the same suite of invertebrates? Pollen Analysis To identify potential pollinators of D. muscipula—defined Methods here as taxa that carry D. muscipula pollen on their bodies— fl Study System we sampled and stained pollen from each ower visitor by swabbing all surfaces of its body with a small piece of fuchsin Dionaea muscipula is native only to southeastern North gel (Kearns and Inouye 1993). Hereafter, we refer to the pol- Carolina and northeastern South Carolina, where it grows len swabbed off the insects as the pollen load, and we as- in wet longleaf (Pinus palustris) and pond pine (Pinus sero- sumed that pollen on the insect’s body was available for tina) savannas and the ecotone between evergreen shrub transfer to another plant. We mounted the pollen by melting bogs and dry pine stands (Luken 2005; Schafale 2012). the jelly on a microscope slide and sealing it with a coverslip. fi These habitats have frequent re return intervals and wa- Similarly, we prepared slides of reference pollen from an- terlogged, low-nutrient soils. Because of these conditions, thers and pollinia. We assessed all flower visitor slides under D. muscipula captures prey to satisfy nutrient requirements a compound microscope for the presence or absence of and co-occurs with a diverse community of other carnivo- D. muscipula pollen and heterospecific pollen by compari- rous plants (Ellison and Gotelli 2001). This habitat is well son to the reference pollen. Dionaea muscipula pollen is represented in Pender County, North Carolina, where our large (∼100 µm diameter) and easily distinguished from that study took place. of coflowering species. For the subset of flower visitors that carried D. muscipula pollen, we subjected pollen slides from the 10 most abun- Sampling dant taxa to more detailed analysis. We counted a sample We sampled D. muscipula flower visitors and prey from of pollen grains from up to 10 randomly selected slides per three sites, separated by 2.5–3.7 km. Each site comprised taxon (or all available slides if fewer than 10 individuals car- hundreds to thousands of flowering stems. We sampled on ried pollen). At #200 magnification, we counted the num- four dates in 2016 during peak D. muscipula bloom (May 17, ber of grains of D. muscipula pollen and heterospecific pollen May 24, June 2, and June 10). We collected flower visitors in five fields of view per slide (one in the center of the stained between 1000 and 1820 hours during fair weather for a total sample and four peripheral fields of view from the center of 29.5 person-hours of collecting (68–670 person-minutes sampling point). The final sample count was the sum of per site each time a site was visited). During sampling, we the number of pollen grains in all five fields of view. searched the site and collected all arthropods observed on anthers, stigmas, and flower petals. Arthropods were collected Data Analysis directly into individual vials or netted and transferred into vials, and then they were sacrificed and transported on dry All analyses were conducted using R v.3.3.1 within R studio ice. We regularly washed nets with 70% ethanol to avoid pol- v.1.0.136 (R Development Core Team 2013; RStudio Team len contamination. To develop a local pollen library, we re- 2015). To estimate the number of taxa that might be de- moved a sample of anthers (or pollinia) from D. muscipula tected at flowers and traps with further sampling, we com- and all nearby flowering species and transported them in puted the Chao1 asymptotic richness estimator in the iNEXT vials on dry ice.