First published online March 13, 2003; 10.1104/pp.102.018176
Plant Physiol, April 2003, Vol. 131, pp. 1877-1893
Molecular Interactions between the Specialist Herbivore
Manduca sexta (Lepidoptera, Sphingidae) and Its Natural
Host Nicotiana attenuata: V. Microarray Analysis and
Further Characterization of Large-Scale Changes in Herbivore-Induced
mRNAs1
Dequan
Hui,
Javeed
Iqbal,
Katja
Lehmann,
Klaus
Gase,
Hans Peter
Saluz, and
Ian T.
Baldwin*
Department of Molecular Ecology, Max Planck Institute for Chemical
Ecology, Winzerlaer Strasse 10, D-07745 Jena, Germany (D.H., K.G.,
I.T.B.); and Department of Cell and Molecular Biology,
Hans-Knöll-Institute for Natural Product, Beutenberg Strasse 11a,
D-07745 Jena, Germany (J.I., H.P.S.)
We extend our analysis of the transcriptional
reorganization that occurs when the native tobacco, Nicotiana
attenuata, is attacked by Manduca sexta larvae
by cloning 115 transcripts by mRNA differential display reverse
transcription-polymerase chain reaction and subtractive hybridization
using magnetic beads (SHMB) from the M.
sexta-responsive transcriptome. These transcripts were spotted
as cDNA with eight others, previously confirmed to be differentially
regulated by northern analysis on glass slide microarrays, and
hybridized with Cy3- and Cy5-labeled probes derived from plants after
2, 6, 12, and 24 h of continuous attack. Microarray analysis
proved to be a powerful means of verifying differential expression; 73 of the cloned genes (63%) were differentially regulated (in equal
proportions from differential display reverse transcription-polymerase chain reaction and SHMB procedures), and of these, 24 (32%) had similarity to known genes or putative proteins (more from SHMB). The
analysis provided insights into the signaling and transcriptional basis
of direct and indirect defenses used against herbivores, suggesting
simultaneous activation of salicylic acid-, ethylene-, cytokinin-,
WRKY-, MYB-, and oxylipin-signaling pathways and implicating terpenoid-, pathogen-, and cell wall-related transcripts in defense responses. These defense responses require resources that could be made
available by decreases in four photosynthetic-related transcripts,
increases in transcripts associated with protein and nucleotide
turnover, and increases in transcripts associated with carbohydrate
metabolism. This putative up-regulation of defense-associated and
down-regulation of growth-associated transcripts occur against a
backdrop of altered transcripts for RNA-binding proteins, putative ATP/ADP translocators, chaperonins, histones, and water channel proteins, responses consistent with a major metabolic reconfiguration that underscores the complexity of response to herbivore attack.
1
This work was supported by the Max Planck Gesellschaft.
*
Corresponding author; e-mail Baldwin{at}ice.mpg.de; fax
49-0-3641-571102.
© 2003 American Society of Plant Biologists
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