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Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway.

ABSTRACT: The sex pheromone biosynthetic pathways of lepidopterans require the concerted actions of multiple gene products. A number of pheromone gland (PG)-specific genes have been cloned in recent years and, whereas in vitro characterizations have indicated functions consistent with roles in pheromone production, there have been no clear demonstrations in vivo. Using an RNA interference-mediated loss-of-function approach, we injected newly formed Bombyx mori pupae with dsRNAs corresponding to genes of interest [i.e., PG fatty acyl reductase (pgFAR), B. mori PG Z11/Delta10,12 desaturase (Bmpgdesat1), PG acyl-CoA-binding protein (pgACBP), midgut ACBP, and pheromone biosynthesis activating neuropeptide receptor (PBANR)] to assess their specific roles during pheromonogenesis. In all cases, the introduced dsRNAs induced a dose-dependent reduction in sex pheromone production with the corresponding decrease in transcript levels. No effects on pupal development or adult emergence were observed. Disrupting the PBANR gene resulted in a loss of the lipase activity that liberates pheromone precursors, whereas knockout of the pgACBP gene prevented the daily accumulation and fluctuation of the triacylglycerols that function as the cellular deposits for the pheromone precursors. Taken together, our results provide unequivocal evidence that the pgACBP, Bmpgdesat1, pgFAR, and PBANR gene products are essential during pheromonogenesis and demonstrate the power of this methodology for dissecting the molecular interactions that comprise biosynthetic pathways.

SUBMITTER: Ohnishi A

PROVIDER: S-EPMC1450183 | biostudies-literature | 2006 Mar

REPOSITORIES: biostudies-literature

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Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway.

Ohnishi Atsushi A Hull J Joe JJ Matsumoto Shogo S

Proceedings of the National Academy of Sciences of the United States of America 20060314 12

The sex pheromone biosynthetic pathways of lepidopterans require the concerted actions of multiple gene products. A number of pheromone gland (PG)-specific genes have been cloned in recent years and, whereas in vitro characterizations have indicated functions consistent with roles in pheromone production, there have been no clear demonstrations in vivo. Using an RNA interference-mediated loss-of-function approach, we injected newly formed Bombyx mori pupae with dsRNAs corresponding to genes of i ...[more]

PMID: 16537410

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Project description:In insects and other animals, intraspecific communication between individuals of the opposite sex is mediated in part by chemical signals called sex pheromones. In most moth species, male moths rely heavily on species-specific sex pheromones emitted by female moths to identify and orient towards an appropriate mating partner among a large number of sympatric insect species. The silkmoth, Bombyx mori, utilizes the simplest possible pheromone system, in which a single pheromone component, (E, Z)-10,12-hexadecadienol (bombykol), is sufficient to elicit full sexual behavior. We have previously shown that the sex pheromone receptor BmOR1 mediates specific detection of bombykol in the antennae of male silkmoths. However, it is unclear whether the sex pheromone receptor is the minimally sufficient determination factor that triggers initiation of orientation behavior towards a potential mate. Using transgenic silkmoths expressing the sex pheromone receptor PxOR1 of the diamondback moth Plutella xylostella in BmOR1-expressing neurons, we show that the selectivity of the sex pheromone receptor determines the chemical response specificity of sexual behavior in the silkmoth. Bombykol receptor neurons expressing PxOR1 responded to its specific ligand, (Z)-11-hexadecenal (Z11-16:Ald), in a dose-dependent manner. Male moths expressing PxOR1 exhibited typical pheromone orientation behavior and copulation attempts in response to Z11-16:Ald and to females of P. xylostella. Transformation of the bombykol receptor neurons had no effect on their projections in the antennal lobe. These results indicate that activation of bombykol receptor neurons alone is sufficient to trigger full sexual behavior. Thus, a single gene defines behavioral selectivity in sex pheromone communication in the silkmoth. Our findings show that a single molecular determinant can not only function as a modulator of behavior but also as an all-or-nothing initiator of a complex species-specific behavioral sequence.

Dataset Information

Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway.

Publications

Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway.

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