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Development and validation of a liquid chromatography-electrospray ionization-time-of-flight mass spectrometry method for induced changes in Nicotiana attenuata leaves during simulated herbivory

ABSTRACT: Metabolite profiling of herbivory-induced changes in Nicotiana attenuata was used to construct a correlation-weighted network to visualize intra- and inter-compound spectra relationships from individual mass features, and to annotate 12 major network hubs, including 4 O-acyl sugars and 6 17-hydroxygeranyllinalool diterpene glycosides, metabolites with recently uncovered anti-herbivore functions.

INSTRUMENT(S): micrOTOF II ESI TOF (Bruker)

SUBMITTER: Steffen Neumann

PROVIDER: MTBLS10 | MetaboLights | 2012-08-14

REPOSITORIES: MetaboLights

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			Action	DRS
	MTBLS10	Other
	FILES	Other
	a_MTBLS10_metabolite_profiling_mass_spectrometry.txt	Txt
	i_Investigation.txt	Txt
	m_MTBLS10_metabolite_profiling_mass_spectrometry_v2_maf.tsv	Tabular

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Development and validation of a liquid chromatography-electrospray ionization-time-of-flight mass spectrometry method for induced changes in Nicotiana attenuata leaves during simulated herbivory.

Gaquerel Emmanuel E Heiling Sven S Schoettner Matthias M Zurek Gabriela G Baldwin Ian T IT

Journal of agricultural and food chemistry 20100901 17

A liquid chromatography-electrospray ionization-time-of-flight mass spectrometry (HPLC/ESI-TOF-MS) procedure was developed to characterize changes induced in Nicotiana attenuata leaves 1 h and 5 days after wounding and application of Manduca sexta elicitors. The constancy of the measurement conditions was first confirmed for 22 selected analytes spanning the entire chromatogram. Using the Profile Analysis software, we extracted 367 buckets, which were analyzed by principal component analysis and ...[more]

PMID: 20701244

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Evidence for an evolutionary hourglass pattern in herbivore-induced transcriptomic responses

Project description:Kenetics of herbivore induced responses in Nicotiana attenuata leaves

2017-12-31 | GSE90951 | GEO

Dicer-like proteins and their role in plant-herbivore interactions in Nicotiana attenuata

Project description:DICER-like proteins produce small RNAs that silence genes involved in development and defenses against viruses and pathogens. Which DCLs participate in plant-herbivore interactions remains unstudied. We identified four distinct DCL genes and stably silenced their expression by RNAi in Nicotiana attenuata, a model system for the study of plant-herbivore interactions. Silencing DCL1 expression was lethal to the plants. Manduca sexta larvae performed significantly better on ir-dcl3and ir-dcl4 plants, but not on ir-dcl2 plants compared to wild type plants. Phytohormones, defense metabolites and microarray analyses revealed that when DCL3 and DCL4 were silenced separately, herbivore resistance traits were regulated in distinctly different ways. Crossing of the lines revealed complex interactions in the patterns of regulation. Single ir-dcl4 and double ir-dcl2/ ir-dcl3 plants were impaired in JA accumulation, while JA-Ile was increased in ir-dcl3 plants. Ir-dcl3 and ir-dcl4 plants were impaired in nicotine accumulation; silencing DCL2 in combination with either DCL3 or DCL4 restored nicotine levels to those of WT. Trypsin proteinase inhibitor activity and transcripts were only silenced in ir-dcl3 plants. We conclude that DCL2/3/4 interact in a complex manner to regulate anti-herbivore defenses and that these interactions significantly complicate the already challenging task of understanding smRNA function in the regulation of biotic interactions. Gene expression in leaves of Nicotiana attenuata wild type, irDCL3 and irDCL4 plants was measured at 1 hour after elicitation with oral secretions of Manduca sexta larvae. Three independent experiments were performed with wild type plants and three independent experiments were performed with irDCL3 and irDCL4 plants. A total of 811 genes were identified as differentially regulated in irDCL3 and irDCL4 compared to wild type plants.

2013-12-24 | E-GEOD-30124 | biostudies-arrayexpress

SuperSAGE analysis of the Nicotiana attenuata transcriptome after wounding

Project description:Fatty acid-amino acid conjugates (FACs) in the oral secretion (OS) of the Manduca sexta larvae are necessary and sufficient to elicit the herbivory-specific responses in Nicotiana attenuata, an annual wild tobacco species. We used SuperSAGE combined with 454 sequencing to quantify the early transcriptional changes elicited by the FAC N-linolenoyl-glutamic acid (18:3-Glu) in N. attenuata. The second fully expanded leaf of rosette stage plants were wounded by rolling a fabric pattern wheel three times on each side of the midvein and the wounds were immediately supplied with either 20 uL of 0.02% (v/v) Tween-20/water (solvent control) or 10 uL of synthetic N-linolenoyl-glutamic acid (18:3-Glu; 0.03 nmol/ÂµL in 0.02% (v/v) Tween-20/water). Tissue was collected after 30 min of the treatments and frozen in liquid nitrogen. Total RNA was extracted by the phenol/chloroform-LiCl method and poly(A)+-RNA was purified from total RNA. Subsequent steps for the construction of the SuperSAGE libraries and 454 sequencing were performed as previously described using double NlaIII digestions (Molina et al., 2008, BMC Genomics, 9:553). Two SuperSAGE libraries were generated after wounding and FAC elicitation of Nicotiana attenuata leaves. Leaves were harvested after 30 min of the treatments, total RNA extracted, polyA+ mRNA isolated and used for cDNA synthesis and SuperSAGE analysis as described my Matsumura et al (2003).

2010-10-05 | E-GEOD-18595 | biostudies-arrayexpress

NaMYB8 regulates distinct, optimally distributed herbivore defense traits

Project description:Identifying whether NaMYB8 regulated genes in Nicotiana attenuata

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Characterization of Nicotiana attenuata plants with reduced expression of HSPRO (ir-hspro plants).

Project description:Identification of transcripts that change expression in roots of Nicotiana attenuata plants with reduced expression of HSPRO and in association with Piriformospora indica. Gene expression in roots of Nicotiana attenuata wild type and ir-hspro seedlings was measured at 14 days after inoculation with Piriformospora indica. Three independent experiments were performed with wild type plants and three independent experiments were performed with ir-hspro plants.

2012-11-06 | E-GEOD-35086 | biostudies-arrayexpress

Characterization of Nicotiana attenuata plants with reduced expression of HD-Zip HD20 (ir-hd20 plants).

Project description:Identification of transcripts that change expression in corollas of Nicotiana attenuata plants with reduced expression of the transcription factor HD20. Gene expression in opening corollas of Nicotiana attenuata wild type and ir-hd20 plants was measured from isolated corollas collected at 12 pm. Three independent experiments were performed with wild type plants and three independent experiments were performed with ir-hd20 plants. A total of 274 genes were identified as differentially regulated in ir-hd20 plants compared to wild type plants.

2013-12-24 | E-GEOD-33682 | biostudies-arrayexpress

NaMYC2 transcription factor regulates a subset of plant defense responses in Nicotiana attenuata

Project description:Identification of significantly altered transcripts in Nicotiana attenuata plants with reduced MYC2 expression after induction with oral secretions of Manduca sexta larvae. Gene expression in leaves of Nicotiana attenuata EV and MYC2_VIGS plants was measured at 1 hour after induction with oral secretions of Manduca sexta larvae. Three independent experiments were performed with EV plants and three independent experiments were performed with MYC2_VIGS plants. A total of 47 genes were identified as differentially regulated in MYC2_VIGS plants compared to EV plants.

2013-05-17 | E-GEOD-45608 | biostudies-arrayexpress

TOC1 silencing in roots only changes transcriptional responses to drought in Nicotiana attenuata

Project description:Identification of transcripts that change expression in TOC1-silenced Nicotiana attenuata plants leaves and roots under drought

2019-01-08 | GSE124736 | GEO

JAZi-mediated jasmonate signaling is invoved in floral defense in Nicotiana attenuata

Project description:Identification of transcripts that change expression in Nicotiana attenuata plants with reduced expression of JAZi in flowers

2017-07-12 | GSE101143 | GEO

Characterization of Nicotiana attenuata plants with reduced expression of lectin receptor kinase 1 (ir-lecRK1 plants).

Project description:Identification of transcripts that change expression in Nicotiana attenuata plants with reduced expression of lectin receptor kinase 1 after elicitation with oral secretions of Manduca sexta larvae. Gene expression in leaves of Nicotiana attenuata wild type and ir-lecRK1 plants was measured at 1 hour after elicitation with oral secretions of Manduca sexta larvae. Three independent experiments were performed with wild type plants and three independent experiments were performed with ir-lecRK1 plants. A total of 148 genes were identified as differentially regulated in ir-lecRK1 plants compared to wild type plants.

2011-11-23 | E-GEOD-29905 | biostudies-arrayexpress

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