Pyrosequencing the Manduca sexta larval midgut transcriptome: messages for digestion, detoxification and defence

Y Pauchet, P Wilkinson, H Vogel, D R Nelson, S E Reynolds, D G Heckel, R H Ffrench-Constant

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

The tobacco hornworm Manduca sexta is an important model for insect physiology but genomic and transcriptomic data are currently lacking. Following a recent pyrosequencing study generating immune related expressed sequence tags (ESTs), here we use this new technology to define the M. sexta larval midgut transcriptome. We generated over 387 000 midgut ESTs, using a combination of Sanger and 454 sequencing, and classified predicted proteins into those involved in digestion, detoxification and immunity. In many cases the depth of 454 pyrosequencing coverage allowed us to define the entire cDNA sequence of a particular gene. Many new M. sexta genes are described including up to 36 new cytochrome P450s, some of which have been implicated in the metabolism of host plant-derived nicotine. New lepidopteran gene families such as the beta-fructofuranosidases, previously thought to be restricted to Bombyx mori, are also described. An unexpectedly high number of ESTs were involved in immunity, for example 39 contigs encoding serpins, and the increasingly appreciated role of the midgut in insect immunity is discussed. Similar studies of other tissues will allow for a tissue by tissue description of the M. sexta transcriptome and will form an essential complimentary step on the road to genome sequencing and annotation.
Original languageEnglish
Pages (from-to)61-75
Number of pages15
JournalInsect Molecular Biology
Volume19
Issue number1
DOIs
Publication statusPublished - Feb 2010

Fingerprint

Manduca
Manduca sexta
Transcriptome
transcriptome
midgut
Digestion
digestion
Expressed Sequence Tags
expressed sequence tags
Immunity
Insects
immunity
insect immunity
insect physiology
Genes
Serpins
application coverage
Bombyx
nicotine
Cytochromes

Keywords

  • beta-fructofuranosidase
  • expressed sequence tag
  • Manduca sexta
  • cytochrome P450
  • midgut
  • 454 pyrosequencing
  • insect immunity

Cite this

Pyrosequencing the Manduca sexta larval midgut transcriptome: messages for digestion, detoxification and defence. / Pauchet, Y; Wilkinson, P; Vogel, H; Nelson, D R; Reynolds, S E; Heckel, D G; Ffrench-Constant, R H.

In: Insect Molecular Biology, Vol. 19, No. 1, 02.2010, p. 61-75.

Research output: Contribution to journalArticle

Pauchet, Y ; Wilkinson, P ; Vogel, H ; Nelson, D R ; Reynolds, S E ; Heckel, D G ; Ffrench-Constant, R H. / Pyrosequencing the Manduca sexta larval midgut transcriptome: messages for digestion, detoxification and defence. In: Insect Molecular Biology. 2010 ; Vol. 19, No. 1. pp. 61-75.
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