Multidimensional ecological filtering shapes a specialized yet potentially flexible pollination interaction
Abstract
Climate change may not only disrupt plant-pollinator interactions by eliminating either partner, but also disturb the simultaneous alignment of phenology, spatial overlap, signal perception, behavior, and morphological fit necessary for successful pollen transfer. However, the ecological dimensions that make specialized pollination systems vulnerable have rarely been comprehensively assessed within the same system. Here, we combined more than a decade of field observations with floral volatile analysis, antennal electrophysiology, behavioral assays, seasonal population monitoring, quantitative morphology, and in situ introduction experiments to examine the rewardless alpine orchid Cypripedium bardolphianum. Female winter-morph Drosophila immigrans dominated effective pollination during the orchid's short flowering season. The orchid attracted drosophilid flies with ethyl tiglate (ET), a multifunctional volatile associated with aggregation, feeding, and oviposition, but responsiveness to this compound extended across several drosophilid lineages. Temporal and spatial overlap, seasonal phenotype, sex-specific preference, and morphological fit progressively narrowed this broad response pool to the realized pollination niche. Two species that are geographically isolated but are attracted to ET, D. robusta and D. virilis, visited flowers and removed pollinia after in situ introduction, revealing compensatory potential beyond the locally realized interaction. These findings show that specialization can emerge from a narrow multidimensional interaction window even when signal-response breadth is comparatively wide. Therefore, environmental change may disrupt current partners through mismatch, while exposing latent functional compatibility through community reassembly. To predict the persistence of specialized pollination, it is necessary to measure the breadth of signal response together with the number and climate sensitivity of the filters that convert response capacity into pollen transfer.
References
W. S. Armbruster, Floral specialization and angiosperm diversity: phenotypic divergence, fitness trade-offs and realized pollination accuracy. AoB Plants 6, plu003 (2014).
W. S. Armbruster, The specialization continuum in pollination systems: diversity of concepts and implications for ecology, evolution and conservation. Funct. Ecol. 31, 88-100 (2017).
M. Ayasse, F. P. Schiestl, H. F. Paulus, C. Löfstedt, B. Hansson, F. Ibarra, W. Francke, Evolution of reproductive strategies in the sexually deceptive orchid Ophrys sphegodes: How does flower-specific variation of odor signals influence reproductive success? Evolution 54, 1995–2004 (2000).
R. J. Bartelt, A. M. Schaner, L. L. Jackson, Aggregation pheromones in five taxa of the Drosophila virilis species group. Physiol. Entomol. 11, 367-376 (1986).
R. J. Bartelt, L. L. Jackson, A. M. Schaner, Ester components of aggregation pheromone of Drosophila virilis (Diptera: Drosophilidae). J. Chem. Ecol. 11, 1197-1208 (1985).
I. Bartomeus, J. S. Ascher, D. Wagner, B. N. Danforth, S. Colla, S. Kornbluth, R. Winfree, Climate-associated phenological advances in bee pollinators and bee-pollinated plants. Proc. Natl. Acad. Sci. U.S.A. 108, 20645-20649 (2011).
J. Bascompte, M. B. García, R. Ortega, E. L. Rezende, S. Pironon, Mutualistic interactions reshuffle the effects of climate change on plants across the tree of life. Sci. Adv. 5, eaav2539 (2019).
S. Bauer, B. J. Hoye, Migratory animals couple biodiversity and ecosystem functioning worldwide. Science 344, 1242552 (2014).
B. Bohman, R. D. Phillips, M. H. M. Menz, B. W. Berntsson, G. R. Flematti, R. A. Barrow, K. W. Dixon, R. Peakall, Discovery of pyrazines as pollinator sex pheromones and orchid semiochemicals: implications for the evolution of sexual deception. New Phytol. 203, 939-952 (2014).
D. S. Boukal, A. Bideault, B. M. Carreira, A. Sentis, Species interactions under climate change: connecting kinetic effects of temperature on individuals to community dynamics. Curr. Opin. Insect Sci. 35, 88-95 (2019).
J. Brodmann, R. Twele, W. Francke, L. Yi-bo, S. Xi-qiang, M. Ayasse, Orchid mimics honey bee alarm pheromone in order to attract hornets for pollination. Curr. Biol. 19, 1368-1372 (2009).
L. A. Burkle, J. B. Runyon, Drought and leaf herbivory influence floral volatiles and pollinator attraction. Glob. Change Biol. 22, 1644-1654 (2016).
L. A. Burkle, J. C. Marlin, T. M. Knight, Plant-pollinator interactions over 120 years: loss of species, co-occurrence, and function. Science 339, 1611-1615 (2013).
H. Bänziger, H. Sun, Y. Luo, Pollination of wild lady slipper orchids Cypripedium yunnanense and C. flavum (Orchidaceae) in south-west China: why are there no hybrids? Bot. J. Linn. Soc. 156, 51-64 (2008).
P. J. CaraDonna, W. K. Petry, R. M. Brennan, J. L. Cunningham, J. L. Bronstein, N. M. Waser, N. J. Sanders, Interaction rewiring and the rapid turnover of plant–pollinator networks. Ecol. Lett. 20, 385-394 (2017).
L. Chittka, N. E. Raine, Recognition of flowers by pollinators. Curr. Opin. Plant Biol. 9, 428-435 (2006).
P. J. Cribb, The Genus Cypripedium. Timber Press; Royal Botanic Gardens, Kew, Portland, OR; Richmond, Surrey, UK (1997).
A. Depetris-Chauvin, D. Galagovsky, I. W. Keesey, B. S. Hansson, S. Sachse, M. Knaden, Evolution at multiple processing levels underlies odor-guided behavior in the genus Drosophila. Curr. Biol. 33, 4771-4785.e7 (2023).
V. Domínguez-Garcia, F. P. Molina, O. Godoy, I. Bartomeus, Interaction network structure explains species’ temporal persistence in empirical plant–pollinator communities. Nat. Ecol. Evol. 8, 423-429 (2024).
G. Farré-Armengol, I. Filella, J. Llusia, J. Peñuelas, Floral volatile organic compounds: Between attraction and deterrence of visitors under global change. Perspect. Plant Ecol. Evol. Syst. 15, 56-67 (2013).
A. Favre, M. Päckert, S. U. Pauls, S. C. Jähnig, D. Uhl, I. Michalak, A. N. Muellner‐Riehl, The role of the uplift of the Qinghai‐Tibetan Plateau for the evolution of Tibetan biotas. Biol. Rev. 90, 236-253 (2015).
J. R. K. Forrest, J. D. Thomson, An examination of synchrony between insect emergence and flowering in Rocky Mountain meadows. Ecol. Monogr. 81, 469-491 (2011).
A. C. Gaskett, E. Conti, F. P. Schiestl, Floral odor variation in two heterostylous species of Primula. J. Chem. Ecol. 31, 1223-1228 (2005).
B. Goldman-Huertas, R. F. Mitchell, R. T. Lapoint, C. P. Faucher, J. G. Hildebrand, N. K. Whiteman, Evolution of herbivory in Drosophilidae linked to loss of behaviors, antennal responses, odorant receptors, and ancestral diet. Proc. Natl. Acad. Sci. U.S.A. 112, 3026-3031 (2015).
M. Gérard, M. Vanderplanck, C. E. Restrepo, E. Baird, Sensory perception and behaviour of insect pollinators under climate change. Nat. Clim. Change 13, 596-598 (2023).
E. P. Gómez-Ruiz, T. E. Lacher, Climate change, range shifts, and the disruption of a pollinator-plant complex. Sci. Rep. 9, 14048 (2019).
J. P. Harmon, B. T. Barton, On their best behavior: how animal behavior can help determine the combined effects of species interactions and climate change. Ann. N.Y. Acad. Sci. 1297, 139-147 (2013).
R. Harrington, I. Woiwod, T. Sparks, Climate change and trophic interactions. Trends Ecol. Evol. 14, 146-150 (1999).
K. Hedlund, R. J. Bartelt, M. Dicke, L. E. M. Vet, Aggregation pheromones of Drosophila immigrans, D. phalerata, and D. subobscura. J. Chem. Ecol. 22, 1835-1844 (1996).
B. Huang, X. Yang, F. Yu, Y. Luo, Y. Tai, Surprisingly high orchid diversity in travertine and forest areas in the Huanglong valley, China, and implications for conservation. Biodivers. Conserv. 17, 2773-2786 (2008).
M. J. Hutchings, K. M. Robbirt, D. L. Roberts, A. J. Davy, Vulnerability of a specialized pollination mechanism to climate change revealed by a 356-year analysis. Bot. J. Linn. Soc. 186, 498-509 (2018).
D. W. Inouye, Effects of climate change on alpine plants and their pollinators. Ann. N.Y. Acad. Sci. 1469, 26-37 (2020).
H. F. Izumitani, Y. Kusaka, S. Koshikawa, M. J. Toda, T. Katoh, Phylogeography of the subgenus Drosophila (Diptera: Drosophilidae): evolutionary history of faunal divergence between the old and the new worlds. PLoS One 11, e0160051 (2016).
H. Jiang, J. J. Kong, H. C. Chen, Z. Y. Xiang, W. P. Zhang, Z. D. Han, P. C. Liao, Y. I. Lee, Cypripedium subtropicum (Orchidaceae) employs aphid colony mimicry to attract hoverfly (Syrphidae) pollinators. New Phytol. 227, 1213-1221 (2020).
S. D. Johnson, Pollination ecotypes and the origin of plant species. Proc. R. Soc. B 292, 20242787 (2025).
S. D. Johnson, K. E. Steiner, Generalization versus specialization in plant pollination systems. Trends Ecol. Evol. 15, 140-143 (2000).
R. R. Junker, Multifunctional and diverse floral scents mediate biotic interactions embedded in communities. in Deciphering Chemical Language of Plant Communication, J. Blande, R. Glinwood, Eds. Springer International Publishing (2016), pp. 257–282.
A. P. Karremans, F. Pupulin, D. Grimaldi, K. K. Beentjes, R. Butôt, G. E. Fazzi, K. Kaspers, J. Kruizinga, P. Roessingh, E. F. Smets, B. Gravendeel, Pollination of Specklinia by nectar-feeding Drosophila: The first reported case of a deceptive syndrome employing aggregation pheromones in Orchidaceae. Ann. Bot. 116, 437-455 (2015).
M. T. Kimura, K. Beppu, Climatic adaptations in the Drosophila immigrans species group: seasonal migration and thermal tolerance. Ecol. Entomol. 18, 141-149 (1993).
K. Kojima, M. T. Kimura, Life history adaptations and stress tolerance of four domestic species of Drosophila. Entomol. Sci. 6, 135-142 (2003).
C. J. van der Kooi, M. Vallejo-Marín, S. D. Leonhardt, Mutualisms and (a)symmetry in plant–pollinator interactions. Curr. Biol. 31, R91-R99 (2021).
G. Kudo, T. Y. Ida, Early onset of spring increases the phenological mismatch between plants and pollinators. Ecology 94, 2311-2320 (2013).
M. Levitan, W. J. Etges, Climate change and recent genetic flux in populations of Drosophila robusta. BMC Evol. Biol. 5, 4 (2005).
P. Li, Y. B. Luo, P. Bernhardt, X. Q. Yang, Y. Kou, Deceptive pollination of the Lady's Slipper Cypripedium tibeticum (Orchidaceae). Plant Syst. Evol. 262, 53-63 (2006).
P. Li, Y. Luo, P. Bernhardt, Y. Kou, H. Perner, Pollination of Cypripedium plectrochilum (Orchidaceae) by Lasioglossum spp. (Halictidae): The roles of generalist attractants versus restrictive floral architecture. Plant Biol. 10, 220-230 (2008).
S. Mansourian, M. C. Stensmyr, The chemical ecology of the fly. Curr. Opin. Neurobiol. 34, 95-102 (2015).
E. L. Marjakangas, B. Dalsgaard, A. Ordonez, Fundamental interaction niches: towards a functional understanding of ecological networks' resilience. Ecol. Lett. 28, e70146 (2025).
F. Martos, M. L. Cariou, T. Pailler, J. Fournel, B. Bytebier, S. D. Johnson, Chemical and morphological filters in a specialized floral mimicry system. New Phytol. 207, 225-234 (2015).
J. Memmott, P. G. Craze, N. M. Waser, M. V. Price, Global warming and the disruption of plant–pollinator interactions. Ecol. Lett. 10, 710-717 (2007).
P. M. Mirol, J. Routtu, A. Hoikkala, R. K. Butlin, Signals of demographic expansion in Drosophila virilis. BMC Evol. Biol. 8, 59 (2008).
H. Mitsui, K. Beppu, M. T. Kimura, Seasonal life cycles and resource uses of flower‐ and fruit‐feeding drosophilid flies (Diptera: Drosophilidae) in central Japan. Entomol. Sci. 13, 60-67 (2010).
L. Mondello, A. Salvatore, P. Q. Tranchida, A. Casilli, P. Dugo, G. Dugo, Reliable identification of pesticides using linear retention indices as an active tool in gas chromatographic–mass spectrometric analysis. J. Chromatogr. A 1186, 430-433 (2008).
R. Muthukrishnan, T. M. Smiley, P. O. Title, A. M. Fudickar, A. E. Jahn, J. A. Lau, Chasing the niche: escaping climate change threats in place, time, and space. Glob. Change Biol. 31, e70167 (2025).
L. Nunney, Drosophila on oranges: colonization, competition, and coexistence. Ecology 71, 1904-1915 (1990).
J. Ollerton, A. Killick, E. Lamborn, S. Watts, M. Whiston, Multiple meanings and modes: on the many ways to be a generalist flower. Taxon 56, 717-728 (2007).
A. Pauw, A bird's-eye view of pollination: biotic interactions as drivers of adaptation and community change. Annu. Rev. Ecol. Evol. Syst. 50, 477-502 (2019).
R. Peakall, D. Ebert, J. Poldy, R. A. Barrow, W. Francke, C. C. Bower, F. P. Schiestl, Pollinator specificity, floral odour chemistry and the phylogeny of Australian sexually deceptive Chiloglottis orchids: Implications for pollinator-driven speciation. New Phytol. 188, 437-450 (2010).
R. W. Pemberton, Pollination of slipper orchids (Cypripedioideae): A review. Lankesteriana 13, 65–73 (2013).
C. Krause Pham, A. Ray, Conservation of olfactory avoidance in Drosophila Species and identification of repellents for Drosophila suzukii. Sci. Rep. 5, 11527 (2015).
R. D. Phillips, R. Peakall, T. van der Niet, S. D. Johnson, Niche perspectives on plant–pollinator interactions. Trends Plant Sci. 25, 779-793 (2020).
J. G. Rodger, J. M. Bennett, M. Razanajatovo, T. M. Knight, M. van Kleunen, T. Ashman, J. A. Steets, C. Hui, G. Arceo-Gómez, M. Burd, L. A. Burkle, J. H. Burns, W. Durka, L. Freitas, J. E. Kemp, J. Li, A. Pauw, J. C. Vamosi, M. Wolowski, J. Xia, A. G. Ellis, Widespread vulnerability of flowering plant seed production to pollinator declines. Sci. Adv. 7, eabd3524 (2021).
M. J. Ryan, Sexual selection, receiver biases, and the evolution of sex differences. Science 281, 1999-2003 (1998).
C. C. Salzmann, A. M. Nardella, S. Cozzolino, F. P. Schiestl, Variability in floral scent in rewarding and deceptive orchids: the signature of pollinator-imposed selection? Ann. Bot. 100, 757-765 (2007).
V. L. Scaven, N. E. Rafferty, Physiological effects of climate warming on flowering plants and insect pollinators and potential consequences for their interactions. Curr. Zool. 59, 418-426 (2013).
F. P. Schiestl, P. M. Schlüter, Floral isolation, specialized pollination, and pollinator behavior in orchids. Annu. Rev. Entomol. 54, 425-446 (2009).
F. P. Schiestl, S. Dötterl, The evolution of floral scent and olfactory preferences in pollinators: Coevolution or pre-existing bias? Evolution 66, 2042-2055 (2012).
O. Schweiger, J. C. Biesmeijer, R. Bommarco, T. Hickler, P. E. Hulme, S. Klotz, I. Kühn, M. Moora, A. Nielsen, R. Ohlemüller, T. Petanidou, S. G. Potts, P. Pyšek, J. C. Stout, M. T. Sykes, T. Tscheulin, M. Vilà, G. R. Walther, C. Westphal, M. Winter, M. Zobel, J. Settele, Multiple stressors on biotic interactions: how climate change and alien species interact to affect pollination. Biol. Rev. 85, 777-795 (2010).
C. Simon, F. Frati, A. Beckenbach, B. Crespi, H. Liu, P. Flook, Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Ann. Entomol. Soc. Am. 87, 651-701 (1994).
J. Stökl, A. Strutz, A. Dafni, A. Svatos, J. Doubsky, M. Knaden, S. Sachse, B. S. Hansson, M. C. Stensmyr, A deceptive pollination system targeting drosophilids through olfactory mimicry of yeast. Curr. Biol. 20, 1846-1852 (2010).
K. Suetsugu, S. K. Hirota, N. Okui, Y. Okuyama, M. T. Kimura, Drosophilid pollination in mycoheterotrophic orchids reveals a brood-site deception–mutualism continuum and phylogenetic conservatism. J. Exp. Bot. 77, 4679-4694 (2026).
A. Suwito, H. Watabe, Discovery of the Drosophila (Drosophila) robusta species group (Diptera, Drosophilidae) from Southeast Asian tropics, with the descriptions of six new species. Entomol. Sci. 13, 381-391 (2010).
R. Core Team, R: A language and environment for statistical computing (R Foundation for Statistical Computing, Vienna, Austria, 2026).
L. H. Throckmorton, The phylogeny, ecology and geography of Drosophila. in Handbook of Genetics, R. C. King, Ed. (Plenum Press, New York (1975), vol. 3, pp. 421–469.
R. L. Tremblay, J. D. Ackerman, J. K. Zimmerman, R. N. Calvo, Variation in sexual reproduction in orchids and its evolutionary consequences: a spasmodic journey to diversification. Biol. J. Linn. Soc. 84, 1-54 (2004).
J. Trunschke, R. R. Junker, G. Kudo, J. M. Alexander, S. K. Richman, I. Till-Bottraud, Effects of climate change on plant-pollinator interactions and its multitrophic consequences. Alpine Bot. 134, 115-121 (2024).
T. Tully, W. G. Quinn, Classical conditioning and retention in normal and mutant Drosophila melanogaster. J. Comp. Physiol. A 157, 263–277 (1985).
T. Ueno, A. Takenoshita, K. Hamamichi, M. P. Sato, Y. Takahashi, Rapid seasonal changes in phenotypes in a wild Drosophila population. Sci. Rep. 13, 21940 (2023).
B. Wang, Z. Tong, Y. Xiong, X. Wang, W. S. Armbruster, S. Huang, The evolution of flower–pollinator trait matching, and why do some alpine gingers appear to be mismatched? Ann. Bot. 132, 1073-1088 (2023).
H. Watabe, X. C. Liang, W. X. Zhang, The Drosophila robusta species-group (Diptera: Drosophilidae) from Yunnan Province, southern China, with the revision of its geographic distribution. Zool. Sci. 7, 133–140 (1990).
H. Wickham, ggplot2: Elegant graphics for data analysis (Springer-Verlag, New York, 2016).
A. H. Yamamoto, Temperature preference of Drosophila immigrans and D. virilis: Intra- and inter-population genetic variation. Jpn. J. Genet. 69, 67-76 (1994).
S. He, T. Wang, Y.J. Yu, X.N. Yang, W.F. Zhang, B.Y. Lu, Y.B. Luo, Ovipositor morphology and mechanosensory innovation traits favor niche width expansion in Drosophila. bioRxiv 2026.04.29.721748v2 [Preprint] (2026). doi:10.64898/2026.04.29.721748.
P. L. Zarnetske, D. K. Skelly, M. C. Urban, biotic multipliers of climate change. Science 336, 1516-1518 (2012).
W. F. Zhang, L. M. Liu, S. He, B. Y. Lu, Y. B. Luo, Production and evolution pattern of “fruity smell” aggregation pheromones in genus Drosophila. J. Syst. Evol. 60, 208-219 (2022).
W. Zhang, Y. Chen, Z. Dai, Mitochondrial DNA genetic polymorphism of Drosophila immigrans in China. Acta Genet. Sin. 26, 336–344 (1999).
W. Zhang, Y. Luo, Non-associative learning underlies pollination interaction of pollinators and flowering plants. Natl. Sci. Open 3, 20230031 (2024).
L. Zhao, D. J. Begun, Genomics of parallel adaptation at two timescales in Drosophila. PLoS Genet. 13, e1007016 (2017).
C. Zheng, Z. Ren, S. Li, W. Zhang, A. R. Rech, Y. Gao, G. Wang, B. Xu, X. Gao, Pollen-like trichomes may reward pollinators of Cypripedium wardii (Orchidaceae). iScience 28, 113951 (2025).
Metrics
DOI:
Submission ID:
Downloads
Posted
How to Cite
Download Citation
Declaration of Competing Interests
The authors declare no competing interests to disclose.
Copyright
The copyright holder for this preprint is the author/funder.
All rights reserved. This work is protected by copyright. No part of this work may be reproduced, distributed, or transmitted in any form or by any means without the prior written permission of the copyright holder.