SuperSAGE analysis of the Nicotiana attenuata transcriptome after fatty acid-amino acid elicitation (FAC): identification of early mediators of insect responses

BMC Plant Biol. 2010 Apr 14;10:66. doi: 10.1186/1471-2229-10-66.


Background: Plants trigger and tailor defense responses after perception of the oral secretions (OS) of attacking specialist lepidopteran larvae. Fatty acid-amino acid conjugates (FACs) in the OS of the Manduca sexta larvae are necessary and sufficient to elicit the herbivory-specific responses in Nicotiana attenuata, an annual wild tobacco species. How FACs are perceived and activate signal transduction mechanisms is unknown.

Results: We used SuperSAGE combined with 454 sequencing to quantify the early transcriptional changes elicited by the FAC N-linolenoyl-glutamic acid (18:3-Glu) and virus induced gene silencing (VIGS) to examine the function of candidate genes in the M. sexta-N. attenuata interaction. The analysis targeted mRNAs encoding regulatory components: rare transcripts with very rapid FAC-elicited kinetics (increases within 60 and declines within 120 min). From 12,744 unique Tag sequences identified (UniTags), 430 and 117 were significantly up- and down-regulated >or= 2.5-fold, respectively, after 18:3-Glu elicitation compared to wounding. Based on gene ontology classification, more than 25% of the annotated UniTags corresponded to putative regulatory components, including 30 transcriptional regulators and 22 protein kinases. Quantitative PCR analysis was used to analyze the FAC-dependent regulation of a subset of 27 of these UniTags and for most of them a rapid and transient induction was confirmed. Six FAC-regulated genes were functionally characterized by VIGS and two, a putative lipid phosphate phosphatase (LPP) and a protein of unknown function, were identified as important mediators of the M. sexta-N. attenuata interaction.

Conclusions: The analysis of the early changes in the transcriptome of N. attenuata after FAC elicitation using SuperSAGE/454 has identified regulatory genes involved in insect-specific mediated responses in plants. Moreover, it has provided a foundation for the identification of additional novel regulators associated with this process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / metabolism*
  • Animals
  • Databases, Genetic
  • Expressed Sequence Tags
  • Fatty Acids / metabolism*
  • Gene Expression Profiling*
  • Gene Expression Regulation, Plant*
  • Gene Library
  • Gene Silencing
  • Manduca / physiology*
  • Molecular Sequence Data
  • Phosphatidate Phosphatase / metabolism
  • Plant Leaves / parasitology
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reproducibility of Results
  • Tobacco / enzymology
  • Tobacco / genetics*
  • Tobacco / parasitology*


  • Amino Acids
  • Fatty Acids
  • RNA, Messenger
  • lipid phosphate phosphatase
  • Phosphatidate Phosphatase

Associated data

  • GENBANK/GT184388
  • GENBANK/GT184389
  • GENBANK/GT184390
  • GENBANK/GT184391
  • GENBANK/GT184392
  • GENBANK/GT184393
  • GENBANK/GT184394
  • GENBANK/GT184395
  • GENBANK/GT184396
  • GENBANK/GT184397
  • GENBANK/GT184398
  • GENBANK/GT184399
  • GENBANK/GT184400
  • GENBANK/GT184401
  • GENBANK/GT184402
  • GENBANK/GT184403
  • GENBANK/GT184404
  • GENBANK/GT184405
  • GENBANK/GT184406
  • GENBANK/GT184407
  • GENBANK/GT184408
  • GENBANK/GT184409
  • GENBANK/GT184410
  • GENBANK/GT184411
  • GENBANK/GT184412
  • GENBANK/GT184413
  • GEO/GSE18595