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Analysis of the Agrotis segetum pheromone gland transcriptome in the light of sex pheromone biosynthesis

Ding, Bao-Jian LU and Löfstedt, Christer LU (2015) In BMC Genomics 16. p.711-711
Abstract
Background: Moths rely heavily on pheromone communication for mate finding. The pheromone components of most moths are modified from the products of normal fatty acid metabolism by a set of tissue-specific enzymes. The turnip moth, Agrotis segetum uses a series of homologous fatty-alcohol acetate esters ((Z)-5-decenyl, (Z)-7-dodecenyl, and (Z)-9 tetradecenyl acetate) as its sex pheromone components. The ratio of the components differs between populations, making this species an interesting subject for studies of the enzymes involved in the biosynthetic pathway and their influence on sex pheromone variation.

Results: Illumina sequencing and comparative analysis of the transcriptomes of the pheromone gland and

... (More)
Background: Moths rely heavily on pheromone communication for mate finding. The pheromone components of most moths are modified from the products of normal fatty acid metabolism by a set of tissue-specific enzymes. The turnip moth, Agrotis segetum uses a series of homologous fatty-alcohol acetate esters ((Z)-5-decenyl, (Z)-7-dodecenyl, and (Z)-9 tetradecenyl acetate) as its sex pheromone components. The ratio of the components differs between populations, making this species an interesting subject for studies of the enzymes involved in the biosynthetic pathway and their influence on sex pheromone variation.

Results: Illumina sequencing and comparative analysis of the transcriptomes of the pheromone gland and

abdominal epidermal tissue, enabled us to identify genes coding for putative key enzymes involved in the

pheromone biosynthetic pathway, such as fatty acid synthase, β-oxidation enzymes, fatty-acyl desaturases (FAD), fatty-acyl reductases (FAR), and acetyltransferases. We functionally assayed the previously identified Δ11-desaturase

[GenBank: ES583599, JX679209] and FAR [GenBank: JX679210] and candidate acetyltransferases (34 genes) by

heterologous expression in yeast. The functional assay confirmed that the Δ11-desaturase interacts with palmitate and produces (Z)-11-hexadecenoate, which is the common unsaturated precursor of three homologous pheromone component acetates produced by subsequent chain-shortening, reduction and acetylation. Much lower, but still visible, activity on 14C and 12C saturated acids may account for minor pheromone compounds previously observed in

the pheromone gland. The FAR characterized can operate on various unsaturated fatty acids that are the immediate acyl precursors of the different A. segetum pheromone components. None of the putative acetyltransferases that we expressed heterologously did acetylate any of the fatty alcohols tested as substrates.

Conclusions: The massive sequencing technology generates enormous amounts of candidate genes potentially

involved in pheromone biosynthesis but testing their function by heterologous expression or gene silencing is a

bottleneck. We confirmed the function of a previously identified desaturase gene and a fatty-acyl reductase gene

by heterologous expression, but the acetyltransferase postulated to be involved in pheromone biosynthesis remains

illusive, in spite of 34 candidates being assayed. We also generated lists of gene candidates that may be useful for

characterizing the acetyl-CoA carboxylase, fatty acid synthetase and β-oxidation enzymes. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Agrotis segetum, Pheromone biosynthesis, Transcriptome, Desaturase, Fatty-acyl reductase, Acetyltransferase, Functional assay, Yeast expression
in
BMC Genomics
volume
16
pages
711 - 711
publisher
BioMed Central
external identifiers
  • pmid:26385554
  • wos:000361354100003
  • scopus:84961158258
ISSN
1471-2164
DOI
10.1186/s12864-015-1909-2
project
Evolutionary mechanisms of pheromone divergence in Lepidoptera
language
English
LU publication?
yes
id
efaae308-242a-41fc-a4df-a6ab57e7fd5e (old id 7870398)
date added to LUP
2015-09-20 15:51:36
date last changed
2017-10-22 04:12:12
@article{efaae308-242a-41fc-a4df-a6ab57e7fd5e,
  abstract     = {Background: Moths rely heavily on pheromone communication for mate finding. The pheromone components of most moths are modified from the products of normal fatty acid metabolism by a set of tissue-specific enzymes. The turnip moth, <i>Agrotis segetum</i> uses a series of homologous fatty-alcohol acetate esters ((Z)-5-decenyl, (Z)-7-dodecenyl, and (Z)-9 tetradecenyl acetate) as its sex pheromone components. The ratio of the components differs between populations, making this species an interesting subject for studies of the enzymes involved in the biosynthetic pathway and their influence on sex pheromone variation.<br/><br>
Results: Illumina sequencing and comparative analysis of the transcriptomes of the pheromone gland and<br/><br>
abdominal epidermal tissue, enabled us to identify genes coding for putative key enzymes involved in the<br/><br>
pheromone biosynthetic pathway, such as fatty acid synthase, β-oxidation enzymes, fatty-acyl desaturases (FAD), fatty-acyl reductases (FAR), and acetyltransferases. We functionally assayed the previously identified Δ11-desaturase<br/><br>
[GenBank: ES583599, JX679209] and FAR [GenBank: JX679210] and candidate acetyltransferases (34 genes) by<br/><br>
heterologous expression in yeast. The functional assay confirmed that the Δ11-desaturase interacts with palmitate and produces (Z)-11-hexadecenoate, which is the common unsaturated precursor of three homologous pheromone component acetates produced by subsequent chain-shortening, reduction and acetylation. Much lower, but still visible, activity on 14C and 12C saturated acids may account for minor pheromone compounds previously observed in<br/><br>
the pheromone gland. The FAR characterized can operate on various unsaturated fatty acids that are the immediate acyl precursors of the different A. segetum pheromone components. None of the putative acetyltransferases that we expressed heterologously did acetylate any of the fatty alcohols tested as substrates.<br/><br>
Conclusions: The massive sequencing technology generates enormous amounts of candidate genes potentially<br/><br>
involved in pheromone biosynthesis but testing their function by heterologous expression or gene silencing is a<br/><br>
bottleneck. We confirmed the function of a previously identified desaturase gene and a fatty-acyl reductase gene<br/><br>
by heterologous expression, but the acetyltransferase postulated to be involved in pheromone biosynthesis remains<br/><br>
illusive, in spite of 34 candidates being assayed. We also generated lists of gene candidates that may be useful for<br/><br>
characterizing the acetyl-CoA carboxylase, fatty acid synthetase and β-oxidation enzymes.},
  author       = {Ding, Bao-Jian and Löfstedt, Christer},
  issn         = {1471-2164},
  keyword      = {Agrotis segetum,Pheromone biosynthesis,Transcriptome,Desaturase,Fatty-acyl reductase,Acetyltransferase,Functional assay,Yeast expression},
  language     = {eng},
  pages        = {711--711},
  publisher    = {BioMed Central},
  series       = {BMC Genomics},
  title        = {Analysis of the <i>Agrotis segetum</i> pheromone gland transcriptome in the light of sex pheromone biosynthesis},
  url          = {http://dx.doi.org/10.1186/s12864-015-1909-2},
  volume       = {16},
  year         = {2015},
}