Summary of Study ST001489

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001006. The data can be accessed directly via it's Project DOI: 10.21228/M8NT36 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST001489
Study Title	Metabolomics by UHPLC-HRMS reveals the impact of heat stress on pathogen-elicited immunity in maize
Study Summary	Studies investigating crop resistance to biotic and abiotic stress have largely focused on plant responses to singular forms of stress and individual biochemical pathways that only partially represent stress responses. Thus, combined biotic and abiotic stress treatments and the global assessment of their elicited metabolic expression remains largely unexplored. In this study, we employed targeted and untargeted metabolomics to investigate the metabolic responses of maize (Zea mays) to both individual and combinatorial stress treatments using heat (abiotic) and Cochliobolus heterostrophus infection (biotic) experiments. Ultra-high-performance liquid chromatography-high-resolution mass spectrometry revealed significant metabolic responses to C. heterostrophus infection and heat stress, and comparative analyses between these individual forms of stress demonstrated differential elicitation between the two global metabolomes. In combinatorial experiments, treatment with heat stress prior to fungal inoculation negatively impacted maize disease resistance against C. heterostrophus, and distinct metabolome separation between combinatorial stressed plants and the non-heat stressed infected controls was observed. Targeted analysis revealed inducible primary and secondary metabolite responses to biotic/abiotic stress, and combinatorial experiments indicated that deficiency in the hydroxycinnamic acid, p-coumaric acid, may lead to the heat-induced susceptibility of maize to C. heterostrophus. Collectively, these findings demonstrate that abiotic stress can predispose crops to more severe disease symptoms, underlining the increasing need to investigate defense chemistry in plants under combinatorial stress.
Institute	Agricultural Research Service, United States Department of Agriculture
Department	Center of Medical, Agricultural, and Veterinary Entomology
Laboratory	Chemistry Research Unit
Last Name	Christensen
First Name	Shawn
Address	1600 SW 23rd Drive Gainesville, FL 32608
Email	shawn.christensen@usda.gov
Phone	3523745739
Submit Date	2020-08-03
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2021-08-03
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001571
Sampleprep Summary:	LCMS grade reagents (Thermo Fisher Scientific) were used throughout the extraction. Frozen 1.5-mL fast prep tubes with Zirmil® beads (1.1mm; SEPR Ceramic Beads and Powders, Mountainside, NJ, USA) containing 50 mg of ground and frozen tissue were carefully thawed to 4°C then transferred to ice. An internal standard mix of 12 μL containing caffeine, D6-Abscisic acid, D5-Jasmonic acid, D5-Cinnamic acid, D5-Indole-3-acetic acid, 13C-alpha linolenic acid, and nicotine at 8.33 μg/mL was added to each sample along with 750 μL of 10 mM of ammonium acetate and 750 μL of methanol (MeOH). Samples were mixed by vortex then homogenized at 6000 RPM for 30 seconds in a FastPrep® FP 120 tissue homogenizer (Qbiogene) and then sonicated in an ambient water bath for 15 min. Samples were centrifuged for 10 minutes at 20,000 RCF at ambient temperature, then 1 mL of supernatant was transferred to individual 4 mL glass vials. The supernatant was dried under a nitrogen stream at 30°C, reconstituted in 200 μL of 0.1% formic acid in water, and vortexed for 30 seconds. Vial contents were transferred to a 1.5-mL snap-cap tube, placed on ice for 10 minutes, then centrifuged at 20,000 RCF for 10 minutes at ambient temperature. 150 μL of supernatant was transferred to glass LC vials for UHPLC-HRMS analysis.

Sampleprep ID:

SP001571

Sampleprep Summary:

LCMS grade reagents (Thermo Fisher Scientific) were used throughout the extraction. Frozen 1.5-mL fast prep tubes with Zirmil® beads (1.1mm; SEPR Ceramic Beads and Powders, Mountainside, NJ, USA) containing 50 mg of ground and frozen tissue were carefully thawed to 4°C then transferred to ice. An internal standard mix of 12 μL containing caffeine, D6-Abscisic acid, D5-Jasmonic acid, D5-Cinnamic acid, D5-Indole-3-acetic acid, 13C-alpha linolenic acid, and nicotine at 8.33 μg/mL was added to each sample along with 750 μL of 10 mM of ammonium acetate and 750 μL of methanol (MeOH). Samples were mixed by vortex then homogenized at 6000 RPM for 30 seconds in a FastPrep® FP 120 tissue homogenizer (Qbiogene) and then sonicated in an ambient water bath for 15 min. Samples were centrifuged for 10 minutes at 20,000 RCF at ambient temperature, then 1 mL of supernatant was transferred to individual 4 mL glass vials. The supernatant was dried under a nitrogen stream at 30°C, reconstituted in 200 μL of 0.1% formic acid in water, and vortexed for 30 seconds. Vial contents were transferred to a 1.5-mL snap-cap tube, placed on ice for 10 minutes, then centrifuged at 20,000 RCF for 10 minutes at ambient temperature. 150 μL of supernatant was transferred to glass LC vials for UHPLC-HRMS analysis.