Mutagenesis of odorant coreceptor Orco fully disrupts foraging but not oviposition behaviors in the hawkmoth Manduca sexta

Richard A. Fandino*, H. Alexander, S. Bisch-Knaden, J. Zhang, S. Bucks, T.A.T. Nguyen, K. Schröder, Achim Werckenthin, J. Rybak, Monika Stengl, M. Knaden, Bill S. Hansson, Ewald Groβe-Wilde

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

The hawkmoth Manduca sexta and one of its preferred hosts in the North American Southwest, Datura wrightii, share a model insect-plant relationship based on mutualistic and antagonistic life-history traits. D. wrightii is the innately preferred nectar source and oviposition host for M. sexta. Hence, the hawkmoth is an important pollinator while the M. sexta larvae are specialized herbivores of the plant. Olfactory detection of plant volatiles plays a crucial role in the behavior of the hawkmoth. In vivo, the odorant receptor coreceptor (Orco) is an obligatory component for the function of odorant receptors (ORs), a major receptor family involved in insect olfaction. We used CRISPR-Cas9 targeted mutagenesis to knock out (KO) the MsexOrco gene to test the consequences of a loss of OR-mediated olfaction in an insect-plant relationship. Neurophysiological characterization revealed severely reduced antennal and antennal lobe responses to representative odorants emitted by D. wrightii. In a wind-tunnel setting with a flowering plant, Orco KO hawkmoths showed disrupted flight orientation and an ablated proboscis extension response to the natural stimulus. The Orco KO gravid female displayed reduced attraction toward a nonflowering plant. However, more than half of hawkmoths were able to use characteristic odor-directed flight orientation and oviposit on the host plant. Overall, OR-mediated olfaction is essential for foraging and pollination behaviors, but plant-seeking and oviposition behaviors are sustained through additional OR-independent sensory cues.

Original languageEnglish
Pages (from-to)15677-15685
Number of pages9
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number31
DOIs
Publication statusPublished - 30 Jul 2019

Fingerprint

Odorant Receptors
Manduca
Oviposition
Mutagenesis
Smell
Insects
Clustered Regularly Interspaced Short Palindromic Repeats
Datura
Plant Nectar
Pollination
Gene Knockout Techniques
Herbivory
Odorants
Cues
Larva

Keywords

  • CRISPR-Cas9
  • Insect olfaction
  • Insect-plant interactions
  • Manduca sexta
  • Orco

Cite this

Fandino, Richard A. ; Alexander, H. ; Bisch-Knaden, S. ; Zhang, J. ; Bucks, S. ; Nguyen, T.A.T. ; Schröder, K. ; Werckenthin, Achim ; Rybak, J. ; Stengl, Monika ; Knaden, M. ; Hansson, Bill S. ; Groβe-Wilde, Ewald. / Mutagenesis of odorant coreceptor Orco fully disrupts foraging but not oviposition behaviors in the hawkmoth Manduca sexta. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 31. pp. 15677-15685.
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title = "Mutagenesis of odorant coreceptor Orco fully disrupts foraging but not oviposition behaviors in the hawkmoth Manduca sexta",
abstract = "The hawkmoth Manduca sexta and one of its preferred hosts in the North American Southwest, Datura wrightii, share a model insect-plant relationship based on mutualistic and antagonistic life-history traits. D. wrightii is the innately preferred nectar source and oviposition host for M. sexta. Hence, the hawkmoth is an important pollinator while the M. sexta larvae are specialized herbivores of the plant. Olfactory detection of plant volatiles plays a crucial role in the behavior of the hawkmoth. In vivo, the odorant receptor coreceptor (Orco) is an obligatory component for the function of odorant receptors (ORs), a major receptor family involved in insect olfaction. We used CRISPR-Cas9 targeted mutagenesis to knock out (KO) the MsexOrco gene to test the consequences of a loss of OR-mediated olfaction in an insect-plant relationship. Neurophysiological characterization revealed severely reduced antennal and antennal lobe responses to representative odorants emitted by D. wrightii. In a wind-tunnel setting with a flowering plant, Orco KO hawkmoths showed disrupted flight orientation and an ablated proboscis extension response to the natural stimulus. The Orco KO gravid female displayed reduced attraction toward a nonflowering plant. However, more than half of hawkmoths were able to use characteristic odor-directed flight orientation and oviposit on the host plant. Overall, OR-mediated olfaction is essential for foraging and pollination behaviors, but plant-seeking and oviposition behaviors are sustained through additional OR-independent sensory cues.",
keywords = "CRISPR-Cas9, Insect olfaction, Insect-plant interactions, Manduca sexta, Orco",
author = "Fandino, {Richard A.} and H. Alexander and S. Bisch-Knaden and J. Zhang and S. Bucks and T.A.T. Nguyen and K. Schr{\"o}der and Achim Werckenthin and J. Rybak and Monika Stengl and M. Knaden and Hansson, {Bill S.} and Ewald Groβe-Wilde",
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Fandino, RA, Alexander, H, Bisch-Knaden, S, Zhang, J, Bucks, S, Nguyen, TAT, Schröder, K, Werckenthin, A, Rybak, J, Stengl, M, Knaden, M, Hansson, BS & Groβe-Wilde, E 2019, 'Mutagenesis of odorant coreceptor Orco fully disrupts foraging but not oviposition behaviors in the hawkmoth Manduca sexta', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 31, pp. 15677-15685. https://doi.org/10.1073/pnas.1902089116

Mutagenesis of odorant coreceptor Orco fully disrupts foraging but not oviposition behaviors in the hawkmoth Manduca sexta. / Fandino, Richard A.; Alexander, H.; Bisch-Knaden, S.; Zhang, J.; Bucks, S.; Nguyen, T.A.T.; Schröder, K.; Werckenthin, Achim; Rybak, J.; Stengl, Monika; Knaden, M.; Hansson, Bill S.; Groβe-Wilde, Ewald.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 31, 30.07.2019, p. 15677-15685.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Mutagenesis of odorant coreceptor Orco fully disrupts foraging but not oviposition behaviors in the hawkmoth Manduca sexta

AU - Fandino, Richard A.

AU - Alexander, H.

AU - Bisch-Knaden, S.

AU - Zhang, J.

AU - Bucks, S.

AU - Nguyen, T.A.T.

AU - Schröder, K.

AU - Werckenthin, Achim

AU - Rybak, J.

AU - Stengl, Monika

AU - Knaden, M.

AU - Hansson, Bill S.

AU - Groβe-Wilde, Ewald

PY - 2019/7/30

Y1 - 2019/7/30

N2 - The hawkmoth Manduca sexta and one of its preferred hosts in the North American Southwest, Datura wrightii, share a model insect-plant relationship based on mutualistic and antagonistic life-history traits. D. wrightii is the innately preferred nectar source and oviposition host for M. sexta. Hence, the hawkmoth is an important pollinator while the M. sexta larvae are specialized herbivores of the plant. Olfactory detection of plant volatiles plays a crucial role in the behavior of the hawkmoth. In vivo, the odorant receptor coreceptor (Orco) is an obligatory component for the function of odorant receptors (ORs), a major receptor family involved in insect olfaction. We used CRISPR-Cas9 targeted mutagenesis to knock out (KO) the MsexOrco gene to test the consequences of a loss of OR-mediated olfaction in an insect-plant relationship. Neurophysiological characterization revealed severely reduced antennal and antennal lobe responses to representative odorants emitted by D. wrightii. In a wind-tunnel setting with a flowering plant, Orco KO hawkmoths showed disrupted flight orientation and an ablated proboscis extension response to the natural stimulus. The Orco KO gravid female displayed reduced attraction toward a nonflowering plant. However, more than half of hawkmoths were able to use characteristic odor-directed flight orientation and oviposit on the host plant. Overall, OR-mediated olfaction is essential for foraging and pollination behaviors, but plant-seeking and oviposition behaviors are sustained through additional OR-independent sensory cues.

AB - The hawkmoth Manduca sexta and one of its preferred hosts in the North American Southwest, Datura wrightii, share a model insect-plant relationship based on mutualistic and antagonistic life-history traits. D. wrightii is the innately preferred nectar source and oviposition host for M. sexta. Hence, the hawkmoth is an important pollinator while the M. sexta larvae are specialized herbivores of the plant. Olfactory detection of plant volatiles plays a crucial role in the behavior of the hawkmoth. In vivo, the odorant receptor coreceptor (Orco) is an obligatory component for the function of odorant receptors (ORs), a major receptor family involved in insect olfaction. We used CRISPR-Cas9 targeted mutagenesis to knock out (KO) the MsexOrco gene to test the consequences of a loss of OR-mediated olfaction in an insect-plant relationship. Neurophysiological characterization revealed severely reduced antennal and antennal lobe responses to representative odorants emitted by D. wrightii. In a wind-tunnel setting with a flowering plant, Orco KO hawkmoths showed disrupted flight orientation and an ablated proboscis extension response to the natural stimulus. The Orco KO gravid female displayed reduced attraction toward a nonflowering plant. However, more than half of hawkmoths were able to use characteristic odor-directed flight orientation and oviposit on the host plant. Overall, OR-mediated olfaction is essential for foraging and pollination behaviors, but plant-seeking and oviposition behaviors are sustained through additional OR-independent sensory cues.

KW - CRISPR-Cas9

KW - Insect olfaction

KW - Insect-plant interactions

KW - Manduca sexta

KW - Orco

U2 - 10.1073/pnas.1902089116

DO - 10.1073/pnas.1902089116

M3 - Article

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SP - 15677

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

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ER -