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Cited by 9Neurobiology and Insect Physiology ResearchInsect-Plant Interactions and ControlInsect Pheromone Research and Control

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Odorant Receptors Mediating Avoidance of Toxic Mustard Oils in Drosophila melanogaster Are Expanded in Herbivorous Relatives

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Matsunaga, Teruyuki, Reisenman, Carolina E, Goldman-Huertas, Benjamin, Rajshekar, Srivarsha, Suzuki, Hiromu C, Tadres, David, Wong, Joshua, Louis, Matthieu, Ramírez, Santiago R, Whiteman, Noah K·2025·Molecular Biology and Evolution
DOI 10.1093/molbev/msaf164PMC PMC12448936

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Abstract

Plants release defense volatile compounds that can deter herbivores. Among them are electrophilic toxins, such as isothiocyanates from mustard plants, that activate pain receptors by contact (i.e. taste) in many animals, including Drosophila melanogaster. While specialist insects have evolved strategies to tolerate toxicity and use mustard plants as hosts, it is unclear whether nonspecialist insects detect and avoid electrophilic toxins via olfaction. To address this, and to understand if specialized insects co-opted these toxic compounds as host plant olfactory cues, we leveraged closely related drosophilid species, including the microbe-feeding D. melanogaster and Scaptomyza pallida, and the mustard-feeding specialist Scaptomyza flava. In olfactory assays, D. melanogaster exposed to allyl isothiocyanate volatiles were rapidly immobilized, demonstrating the high toxicity of this wasabi-derived compound to nonspecialists. Through single sensillum electrophysiological recordings from olfactory organs and behavioral assays, we identified an olfactory receptor (Or) necessary for volatile detection and behavioral aversion to allyl isothiocyanate in D. melanogaster. RNA-sequencing and heterologous expression revealed that S. flava possess lineage-specific, triplicated homologs of this Or and that each paralog exhibited broadened and distinct sensitivity to isothiocyanate compounds. Using AlphaFold2 modeling, site-directed mutagenesis, and electrophysiological recordings, we identified two critical amino acid substitutions that changed the sensitivity of these paralogs from fruit-derived odors to isothiocyanates in the mustard specialist S. flava. Our findings show that nonspecialists can detect electrophiles via olfaction and that their olfactory systems can rapidly adapt to toxic host plant niches through co-option and duplication of ancestral chemosensory genes with few amino acid changes.

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Research works printed but not linked: 85

  1. Aguilar, 2024, Insights into the evolution of herbivory from a leaf-mining fly, Ecosphere, 15, e4764, 10.1002/ecs2.4764 · DOI
  2. Ahuja, 2010, Defence mechanisms of Brassicaceae: implications for plant-insect interactions and potential for integrated pest management. A review, Agron Sustain Dev, 30, 311, 10.1051/agro/2009025 · DOI
  3. Al-Anzi, 2006, Response of Drosophila to wasabi is mediated by Painless, the fly homolog of mammalian TRPA1/ANKTM1, Curr Biol, 16, 1034, 10.1016/j.cub.2006.04.002 · DOI
  4. Anders, 2015, HTSeq—a Python framework to work with high-throughput sequencing data, Bioinformatics, 31, 166, 10.1093/bioinformatics/btu638 · DOI · Paper page
  5. Auer, 2020, Olfactory receptor and circuit evolution promote host specialization, Nature, 579, 402, 10.1038/s41586-020-2073-7 · DOI
  6. Becher, 2010, Flying the fly: long-range flight behavior of Drosophila melanogaster to attractive odors, J Chem Ecol, 36, 599, 10.1007/s10886-010-9794-2 · DOI
  7. Bell, 2018, Taste and flavor perceptions of glucosinolates, isothiocyanates, and related compounds, Mol Nutr Food Res, 62, e1700990, 10.1002/mnfr.201700990 · DOI
  8. Benton, 2023, Structural screens identify candidate human homologs of insect chemoreceptors and cryptic Drosophila gustatory receptor-like proteins, Elife, 12, e85537, 10.7554/eLife.85537 · DOI
  9. Benton, 2007, An essential role for a CD36-related receptor in pheromone detection in Drosophila, Nature, 450, 289, 10.1038/nature06328 · DOI
  10. Bernays, 1987, Proceedings in life sciences, 159
  11. Bischof, 2007, An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases, Proc Natl Acad Sci U S A, 104, 3312, 10.1073/pnas.0611511104 · DOI
  12. Butterwick, 2018, Cryo-EM structure of the insect olfactory receptor Orco, Nature, 560, 447, 10.1038/s41586-018-0420-8 · DOI
  13. Chen, 2018, Fastp: an ultra-fast all-in-one FASTQ preprocessor, Bioinformatics, 34, i884, 10.1093/bioinformatics/bty560 · DOI · Paper page
  14. Chen, 2020, Recent advances in the genetic basis of taste detection in Drosophila, Cell Mol Life Sci, 77, 1087, 10.1007/s00018-019-03320-0 · DOI
  15. Chou, 1978, Prediction of the secondary structure of proteins from their amino acid sequence, Adv Enzymol Relat Areas Mol Biol, 47, 45, 10.1002/9780470122921.ch2 · DOI
  16. Couto, 2005, Molecular, anatomical, and functional organization of the Drosophila olfactory system, Curr Biol, 15, 1535, 10.1016/j.cub.2005.07.034 · DOI
  17. Crowley-Gall, 2016, Population differences in olfaction accompany host shift in Drosophila mojavensis, Proc Biol Sci, 283, 20161562, 10.1098/rspb.2016.1562 · DOI
  18. Cuellar-Nuñez, 2022, Isothiocyanate-rich extracts from cauliflower (Brassica oleracea var. Botrytis) and radish (Raphanus sativus) inhibited metabolic activity and induced ROS in selected human HCT116 and HT-29 colorectal cancer cells, Int J Environ Res Public Health, 19, 14919, 10.3390/ijerph192214919 · DOI
  19. de Bruyne, 1999, Odor coding in a model olfactory organ: the Drosophila maxillary palp, J Neurosci, 19, 4520, 10.1523/JNEUROSCI.19-11-04520.1999 · DOI · Paper page
  20. Dekker, 2006, Olfactory shifts parallel superspecialism for toxic fruit in Drosophila melanogaster sibling, D. sechellia, Curr Biol, 16, 101, 10.1016/j.cub.2005.11.075 · DOI · Paper page
  21. Del Mármol, 2021, The structural basis of odorant recognition in insect olfactory receptors, Nature, 597, 126, 10.1038/s41586-021-03794-8 · DOI
  22. Dobin, 2013, STAR: ultrafast universal RNA-Seq aligner, Bioinformatics, 29, 15, 10.1093/bioinformatics/bts635 · DOI · Paper page
  23. Dobler, 2012, Community-wide convergent evolution in insect adaptation to toxic cardenolides by substitutions in the Na,K-ATPase, Proc Natl Acad Sci U S A, 109, 13040, 10.1073/pnas.1202111109 · DOI
  24. Dobritsa, 2003, Integrating the molecular and cellular basis of odor coding in the Drosophila antenna, Neuron, 37, 827, 10.1016/S0896-6273(03)00094-1 · DOI
  25. Dweck, 2016, Olfactory channels associated with the Drosophila maxillary palp mediate short- and long-range attraction, Elife, 5, e14925, 10.7554/eLife.14925 · DOI
  26. Dweck, 2020, Molecular logic and evolution of bitter taste in Drosophila, Curr Biol, 30, 17, 10.1016/j.cub.2019.11.005 · DOI
  27. Ferreiro, 2017, Drosophila melanogaster white mutant w1118 undergo retinal degeneration, Front Neurosci, 11, 732, 10.3389/fnins.2017.00732 · DOI
  28. Gloss, 2014, Evolution in an ancient detoxification pathway is coupled with a transition to herbivory in the drosophilidae, Mol Biol Evol, 31, 2441, 10.1093/molbev/msu201 · DOI
  29. Gloss, Evolution of herbivory remodels a Drosophila genome [preprint], bioRxiv 767160, 10.1101/767160 · DOI
  30. Gloss, 10.1101/156299 · DOI
  31. Goldman-Huertas, 2015, 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, 10.1073/pnas.1424656112 · DOI
  32. Gonzalez, 2016, Protocol for heterologous expression of insect odourant receptors in Drosophila, Front Ecol Evol, 4, 24, 10.3389/fevo.2016.00024 · DOI
  33. Hashimoto, 2019, Wasabi versus red imported fire ants: preliminary test of repellency of microencapsulated allyl isothiocyanate against Solenopsis invicta (Hymenoptera: Formicidae) using bait traps in Taiwan, Appl Entomol Zool (Jpn), 54, 193, 10.1007/s13355-019-00613-5 · DOI
  34. Haverkamp, 2018, Combinatorial codes and labeled lines: how insects use olfactory cues to find and judge food, mates, and oviposition sites in complex environments, Front Physiol, 9, 49, 10.3389/fphys.2018.00049 · DOI
  35. Himmel, 2023, Remote homolog detection places insect chemoreceptors in a cryptic protein superfamily spanning the tree of life, Curr Biol, 33, 5023, 10.1016/j.cub.2023.10.008 · DOI · Paper page
  36. Hopkins, 2009, Role of glucosinolates in insect-plant relationships and multitrophic interactions, Annu Rev Entomol, 54, 57, 10.1146/annurev.ento.54.110807.090623 · DOI
  37. Ibanez, 2012, Plant insecticidal toxins in ecological networks, Toxins (Basel), 4, 228, 10.3390/toxins4040228 · DOI
  38. Iorio, 2022, Multi-target effects of ß-caryophyllene and carnosic acid at the crossroads of mitochondrial dysfunction and neurodegeneration: from oxidative stress to microglia-mediated neuroinflammation, Antioxidants (Basel), 11, 1199, 10.3390/antiox11061199 · DOI
  39. Kang, 2010, Analysis of Drosophila TRPA1 reveals an ancient origin for human chemical nociception, Nature, 464, 597, 10.1038/nature08848 · DOI
  40. Katoh, 2013, MAFFT multiple sequence alignment software version 7: improvements in performance and usability, Mol Biol Evol, 30, 772, 10.1093/molbev/mst010 · DOI · Paper page
  41. Kim, 2021, Highly contiguous assemblies of 101 drosophilid genomes, Elife, 10, e66405, 10.7554/eLife.66405 · DOI
  42. Kim, 2010, Drosophila TRPA1 channel mediates chemical avoidance in gustatory receptor neurons, Proc Natl Acad Sci U S A, 107, 8440, 10.1073/pnas.1001425107 · DOI
  43. Kurtovic, 2007, A single class of olfactory neurons mediates behavioural responses to a Drosophila sex pheromone, Nature, 446, 542, 10.1038/nature05672 · DOI
  44. Larsson, 2004, Or83b encodes a broadly expressed odorant receptor essential for Drosophila olfaction, Neuron, 43, 703, 10.1016/j.neuron.2004.08.019 · DOI
  45. Levitt, 1978, Conformational preferences of amino acids in globular proteins, Biochemistry, 17, 4277, 10.1021/bi00613a026 · DOI
  46. Lichtenstein, 1964, Insecticides in nature, naturally occurring insecticides in cruciferous crops, J Agric Food Chem, 12, 158, 10.1021/jf60132a017 · DOI
  47. Lin, 2015, Food odors trigger Drosophila males to deposit a pheromone that guides aggregation and female oviposition decisions, Elife, 4, e08688, 10.7554/eLife.08688 · DOI
  48. Linz, 2013, Host plant-driven sensory specialization in Drosophila erecta, Proc Biol Sci, 280, 20130626, 10.1098/rspb.2013.0626 · DOI
  49. Liu, 2020, The molecular basis of host selection in a crucifer-specialized moth, Curr Biol, 30, 4476, 10.1016/j.cub.2020.08.047 · DOI
  50. Maca, 1972, Czechoslovak species of thegenus Scaptomyza Hardy (Diptera:Drosophilidae) and their bionomics, Acta Entomol Bohemoslov, 69, 119
  51. MacLeod, 1989, Evidence for the occurrence of butyl- and isobutylglucosinolates in seeds of Brassica oleracea, Phytochemistry, 28, 1405, 10.1016/S0031-9422(00)97756-2 · DOI
  52. Mandel, 2018, Behavioral aversion to AITC requires both Painless and dTRPA1 in Drosophila, Front Neural Circuits, 12, 45, 10.3389/fncir.2018.00045 · DOI
  53. Mantel, 1966, Evaluation of survival data and two new rank order statistics arising in its consideration, Cancer Chemother Rep, 50, 163
  54. Martin, 2004, History of an invader, Scaptomyza flava (Fallen, 1823) (Diptera: Drosophilidae), N Z J Zool, 31, 27, 10.1080/03014223.2004.9518356 · DOI
  55. Matsunaga, 2022, Evolution of olfactory receptors tuned to mustard oils in herbivorous Drosophilidae, Mol Biol Evol, 39, 10.1093/molbev/msab362 · DOI
  56. Mirdita, 2022, ColabFold: making protein folding accessible to all, Nat Methods, 19, 679, 10.1038/s41592-022-01488-1 · DOI · Paper page
  57. Mithöfer, 2012, Plant defense against herbivores: chemical aspects, Annu Rev Plant Biol, 63, 431, 10.1146/annurev-arplant-042110-103854 · DOI
  58. Nishimura, 2024, DiGAlign: versatile and interactive visualization of sequence alignment for comparative genomics, Microbes Environ, 39, ME23061, 10.1264/jsme2.ME23061 · DOI
  59. Noge, 2015, 4-Oxo-(E)-2-hexenal produced by Heteroptera induces permanent locomotive impairment in crickets that correlates with free thiol depletion, FEBS Open Bio, 5, 319, 10.1016/j.fob.2015.04.004 · DOI
  60. Oh, 2021, Multisensory interactions regulate feeding behavior in Drosophila, Proc Natl Acad Sci U S A, 118, e2004523118, 10.1073/pnas.2004523118 · DOI
  61. Ohashi, 2015, Novel behavioral assay of wasabi avoidance in Drosophila melanogaster (Diptera: Drosophilidae) using a video tracking system, Appl Entomol Zool, 50, 137, 10.1007/s13355-014-0302-y · DOI
  62. Peláez, 2023, Evolution of chemosensory and detoxification gene families across herbivorous Drosophilidae, G3 (Bethesda), 13, 10.1093/g3journal/jkad133 · DOI
  63. Peláez, 2022, Evolution and genomic basis of the plant-penetrating ovipositor: a key morphological trait in herbivorous Drosophilidae, Proc Biol Sci, 289, 20221938, 10.1098/rspb.2022.1938 · DOI
  64. Rambaut A . FigTree, version 1.4.3. Computer program distributed by the author; 2009 [accessed January 4, 2011] [Internet]. Available from: https://scholar.google.com/citations?view_op=view_citation&hl=en&citation_for_view=JiYPDfoAAAAJ:W5xh706n7nkC.
  65. Ray, 2007, Mechanisms of odor receptor gene choice in Drosophila, Neuron, 53, 353, 10.1016/j.neuron.2006.12.010 · DOI
  66. Reddy, 2004, Interactions of insect pheromones and plant semiochemicals, Trends Plant Sci, 9, 253, 10.1016/j.tplants.2004.03.009 · DOI
  67. Reichstein, 1968, Heart poisons in the monarch butterfly. Some aposematic butterflies obtain protection from cardenolides present in their food plants, Science, 161, 861, 10.1126/science.161.3844.861 · DOI
  68. Reisenman, 2019, Food-derived volatiles enhance consumption in Drosophila melanogaster, J Exp Biol, 222, 10.1242/jeb.202762 · DOI
  69. Scott, 2018, Gustatory processing in Drosophila melanogaster, Annu Rev Entomol, 63, 15, 10.1146/annurev-ento-020117-043331 · DOI · Paper page
  70. Semmelhack, 2009, Select Drosophila glomeruli mediate innate olfactory attraction and aversion, Nature, 459, 218, 10.1038/nature07983 · DOI
  71. Shiraiwa, 2008, Multimodal chemosensory integration through the maxillary palp in Drosophila, PLoS One, 3, e2191, 10.1371/journal.pone.0002191 · DOI
  72. Stamatakis, 2014, RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies, Bioinformatics, 30, 1312, 10.1093/bioinformatics/btu033 · DOI · Paper page
  73. Stensmyr, 2012, A conserved dedicated olfactory circuit for detecting harmful microbes in Drosophila, Cell, 151, 1345, 10.1016/j.cell.2012.09.046 · DOI
  74. Takagi, 2024, Olfactory sensory neuron population expansions influence projection neuron adaptation and enhance odour tracking, Nat Commun, 15, 7041, 10.1038/s41467-024-50808-w · DOI
  75. Tocmo, 2021, Covalent modification of proteins by plant-derived natural products: proteomic approaches and biological impacts, Proteomics, 21, e1900386, 10.1002/pmic.201900386 · DOI
  76. Wang, 2024, Structural basis for odorant recognition of the insect odorant receptor OR-Orco heterocomplex, Science, 384, 1453, 10.1126/science.adn6881 · DOI
  77. War, 2012, Mechanisms of plant defense against insect herbivores, Plant Signal Behav, 7, 1306, 10.4161/psb.21663 · DOI
  78. Whiteman, 2011, Mining the plant-herbivore interface with a leafmining Drosophila of Arabidopsis, Mol Ecol, 20, 995, 10.1111/j.1365-294X.2010.04901.x · DOI
  79. Whiteman, 2012, Genes involved in the evolution of herbivory by a leaf-mining, Drosophilid fly, Genome Biol Evol, 4, 900, 10.1093/gbe/evs063 · DOI
  80. Winde, 2011, Insect herbivore counteradaptations to the plant glucosinolate-myrosinase system, Phytochemistry, 72, 1566, 10.1016/j.phytochem.2011.01.016 · DOI
  81. Wu, 2021, Glucosinolates in Brassica vegetables: characterization and factors that influence distribution, content, and intake, Annu Rev Food Sci Technol, 12, 485, 10.1146/annurev-food-070620-025744 · DOI
  82. Xu, 2005, Drosophila OBP LUSH is required for activity of pheromone-sensitive neurons, Neuron, 45, 193, 10.1016/j.neuron.2004.12.031 · DOI
  83. Yaffe, 2015, Piperine, an alkaloid from black pepper, inhibits growth of human colon cancer cells via G1 arrest and apoptosis triggered by endoplasmic reticulum stress, Mol Carcinog, 54, 1070, 10.1002/mc.22176 · DOI
  84. Yao, 2005, Chemosensory coding by neurons in the coeloconic sensilla of the Drosophila antenna, J Neurosci, 25, 8359, 10.1523/JNEUROSCI.2432-05.2005 · DOI
  85. Zhao, 2024, Structural basis of odor sensing by insect heteromeric odorant receptors, Science, 384, 1460, 10.1126/science.adn6384 · DOI

Metadata

Authors
Matsunaga, T., Reisenman, C. E., Goldman-Huertas, B., Rajshekar, S., Suzuki, H. C., Tadres, D., Wong, J., Louis, M., Ramírez, S. R., & Whiteman, N. K.
Affiliations
  • The University of Tokyo (JP)
  • University of California, Berkeley (US)
  • University of California, Santa Barbara (US)
  • Johns Hopkins University (US)
  • University of California, Davis (US)
Corresponding author
  • Matsunaga, Teruyuki
  • Reisenman, Carolina E
  • Whiteman, Noah K
ORCID
Venue
Molecular Biology and Evolution
Publisher
Oxford University Press (OUP)
Published
2025-07-04
Volume
42
Issue
9
ISSN
  • 0737-4038 (ISSN-L)
  • 1537-1719
DOI
10.1093/molbev/msaf164
PMC
PMC12448936
OpenAlex
W4412030535
Cited by
9
References
2
Citations by year
2026: 6 · 2025: 3
FWCI
3.51
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Biology and Evolution (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
Funders
  • National Institutes of Health (P40OD010949)
  • Japan Society for the Promotion of Science (22K20565)
  • National Institute of General Medical Sciences (R35GM119816, R35GM139653)
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurobiology and Insect Physiology Research
  • Life Sciences › Agricultural and Biological Sciences › Insect Science › Insect-Plant Interactions and Control
  • Life Sciences › Agricultural and Biological Sciences › Insect Science › Insect Pheromone Research and Control
Keywords
olfactory receptors, isothiocyanates, allyl isothiocyanate, Drosophila melanogaster, Scaptomyza flava, olfactory avoidance, host plant adaptation

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