SuperSAGE analysis of the Nicotiana attenuata transcriptome after fatty acid-amino acid elicitation (FAC): identification of early mediators of insect responses.
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.
Gilardoni
Paola A
PA
Max Planck Institute for Chemical Ecology, Department of Molecular Ecology, Jena, Germany.
Schuck
Stefan
S
Jüngling
Ruth
R
Rotter
Björn
B
Baldwin
Ian T
IT
Bonaventure
Gustavo
G
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D030541:Databases, Genetic; D020224:Expressed Sequence Tags; D005227:Fatty Acids / Q000378:metabolism*; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D020868:Gene Silencing; D018402:Manduca / Q000502:physiology*; D008969:Molecular Sequence Data; D010711:Phosphatidate Phosphatase / Q000378:metabolism; D018515:Plant Leaves / Q000469:parasitology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics* / Q000469:parasitology*
D000596:Amino Acids; D005227:Fatty Acids; D012333:RNA, Messenger; C112948:lipid phosphate phosphatase; D010711:Phosphatidate Phosphatase
Plant Physiol. 2003 Apr;131(4):1877-93
12692347
Plant Physiol. 2010 Jan;152(1):96-106
19897603
Science. 1998 May 15;280(5366):1091-4
9582125
Plant Cell. 2002 May;14(5):993-1003
12034892
Plant Physiol. 2008 Mar;146(3):952-64
18223147
Plant Physiol. 2002 Jan;128(1):223-35
11788768
Trends Plant Sci. 2000 May;5(5):199-206
10785665
Plant Mol Biol. 2003 Jun;52(3):643-60
12956533
Plant Cell. 2007 Jul;19(7):2213-24
17630279
Science. 2002 Feb 15;295(5558):1299-301
11847343
Annu Rev Phytopathol. 1992;30:391-418
18647100
Plant Cell. 2008 Jul;20(7):1984-2000
18641266
Plant Cell. 1999 Feb;11(2):273-87
9927644
Proc Natl Acad Sci U S A. 2006 Aug 22;103(34):12935-40
16912118
Plant Cell. 2007 Mar;19(3):1096-122
17400894
J Exp Bot. 2004 Jan;55(395):151-7
14623900
J Chem Ecol. 2001 Mar;27(3):547-68
11441445
Plant Physiol. 2006 Dec;142(4):1621-41
17028148
BMC Plant Biol. 2007 Sep 19;7:49
17877838
Plant Physiol. 2008 Jul;147(3):1347-57
18467450
Plant Cell. 2001 Feb;13(2):303-18
11226187
Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8113-8
9653149
Plant Cell. 1999 Feb;11(2):289-98
9927645
Plant Physiol. 2001 Feb;125(2):711-7
11161028
Plant Physiol. 1995 Jul;108(3):949-960
12228518
Plant Physiol. 2008 Mar;146(3):845-51
18316639
Plant Mol Biol. 1992 Nov;20(4):653-62
1450380
Mol Gen Genet. 2000 Feb;263(1):30-7
10732671
Plant Cell. 2002 Jul;14(7):1557-66
12119374
Plant Mol Biol. 2009 Feb;69(3):337-46
19039666
Plant J. 1999 Dec;20(6):719-26
10652144
Plant J. 2004 May;38(4):650-63
15125771
Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15718-23
14676315
Science. 1995 Oct 20;270(5235):484-7
7570003
Plant J. 2004 May;38(4):639-49
15125770
EMBO J. 1996 Oct 15;15(20):5690-700
8896462
Genome Res. 1997 Oct;7(10):986-95
9331369
J Chem Ecol. 1994 Sep;20(9):2373-91
24242812
BMC Genomics. 2008 Nov 24;9:553
19025623
J Biol Chem. 2001 Jun 8;276(23):20300-8
11278556
Plant Physiol. 2004 Mar;134(3):1181-90
14976235
Plant Physiol. 2001 Feb;125(2):683-700
11161026
Plant Mol Biol. 2002 Sep;50(2):155-66
12175009
Plant Physiol. 2003 Apr;131(4):1894-902
12692348
Plant Cell. 2004 Nov;16(11):3132-47
15494554
Plant J. 2004 Apr;38(1):142-51
15053767
Annu Rev Plant Biol. 2008;59:41-66
18031220
Science. 1997 Feb 7;275(5301):832-4
9012350
J Plant Growth Regul. 2003 Dec;22(4):289-297
14676968
Plant Physiol. 2008 Jan;146(1):32-44
18024554
Cell Microbiol. 2005 Jan;7(1):11-8
15617519
Plant Mol Biol. 2001 Sep;47(1-2):197-206
11554472
Cell Res. 2009 Mar;19(3):307-16
19065152