Identification of chemosensory receptor genes in Manduca sexta and knockdown by RNA interference.
BACKGROUND
Insects detect environmental chemicals via a large and rapidly evolving family of chemosensory receptor proteins. Although our understanding of the molecular genetic basis for Drosophila chemoreception has increased enormously in the last decade, similar understanding in other insects remains limited. The tobacco hornworm, Manduca sexta, has long been an important model for insect chemosensation, particularly from ecological, behavioral, and physiological standpoints. It is also a major agricultural pest on solanaceous crops. However, little sequence information and lack of genetic tools has prevented molecular genetic analysis in this species. The ability to connect molecular genetic mechanisms, including potential lineage-specific changes in chemosensory genes, to ecologically relevant behaviors and specializations in M. sexta would be greatly beneficial.


RESULTS
Here, we sequenced transcriptomes from adult and larval chemosensory tissues and identified chemosensory genes based on sequence homology. We also used dsRNA feeding as a method to induce RNA interference in larval chemosensory tissues.


CONCLUSIONS
We report identification of new chemosensory receptor genes including 17 novel odorant receptors and one novel gustatory receptor. Further, we demonstrate that systemic RNA interference can be used in larval olfactory neurons to reduce expression of chemosensory receptor transcripts. Together, our results further the development of M. sexta as a model for functional analysis of insect chemosensation.
Department of Biological Sciences Barnard College, Columbia University, New York, NY 10027, USA.
Natalie
N
Howlett
Katherine L
KL
Dauber
Aditi
A
Shukla
Brian
B
Morton
John I
JI
Glendinning
Elyssa
E
Brent
Caroline
C
Gleason
Fahmida
F
Islam
Denisse
D
Izquierdo
Sweta
S
Sanghavi
Anika
A
Afroz
Aanam
A
Aslam
Marissa
M
Barbaro
Rebekah
R
Blutstein
Margarita
M
Borovka
Brianna
B
Desire
Ayala
A
Elikhis
Qing
Q
Fan
Katherine
K
Hoffman
Amy
A
Huang
Dominique
D
Keefe
Sarah
S
Lopatin
Samara
S
Miller
Priyata
P
Patel
Danielle
D
Rizzini
Alyssa
A
Robinson
Karimah
K
Rokins
Aneta
A
Turlik
Jennifer H
JH
Mansfield
D000818:Animals; D020451:Contig Mapping; D015723:Gene Library; D018014:Gene Transfer Techniques; D007814:Larva / Q000235:genetics / Q000378:metabolism; D018402:Manduca / Q000145:classification / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D018035:Receptors, Odorant / Q000037:antagonists & inhibitors* / Q000145:classification / Q000378:metabolism; D059467:Transcriptome / Q000235:genetics
D012330:RNA, Double-Stranded; D018035:Receptors, Odorant