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Development of Application Methods of Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (Ir-Maldesi-Msi) of Plants.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Development of Application Methods of Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (Ir-Maldesi-Msi) of Plants./
Author:	Bagley, Michael Caleb.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2021,
Description:	425 p.
Notes:	Source: Dissertations Abstracts International, Volume: 83-05, Section: B.
Contained By:	Dissertations Abstracts International83-05B.
Subject:	Physics. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28747700
ISBN:	9798494445728

Development of Application Methods of Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (Ir-Maldesi-Msi) of Plants.
Bagley, Michael Caleb.

Development of Application Methods of Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (Ir-Maldesi-Msi) of Plants. - Ann Arbor : ProQuest Dissertations & Theses, 2021 - 425 p.

Source: Dissertations Abstracts International, Volume: 83-05, Section: B.

Thesis (Ph.D.)--North Carolina State University, 2021.

This item must not be sold to any third party vendors.

Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (IR-MALDESI-MSI) is a hybrid of matrix assisted laser desorption with electrospray post ionization that enables high resolution spatial information to be combined with the ambient characteristics of electrospray. It has been used to complement other MSI techniques by analyzing samples and biomolecules that are not commonly accessible using the dominant techniques of matrix assisted laser desorption ionization (MALDI) and ESI.The yield fluctuation of artemisinin due to its glandular trichome (GT)-specific biosynthesis has remained an unsolved problem in meeting the global demand for this primary treatment for malaria. Detailed IR-MALDESI image profiling revealed that artemisinin, artemisinic acid, and arteannuin B three metabolites and dihydroartemisinin are localized in non-GT cells of leaves of inbred A. annua plants. Finding that non-GT cells can express the artemisinin biosynthetic pathway led to a revision of the current dogma of artemisinin biosynthesis in A. annua and may expedite innovation of novel metabolic engineering approaches for high and stable production of artemisinin.Quantitative MSI was performed on young Arabidopsis thaliana seedlings to understand how mutations affect metabolite accumulation in plant development. A new method for the quantification of auxin‐related compounds was created using stable‐ isotope‐labelled (SIL) indole‐3‐acetic acid (IAA) doped into agarose substrate. Relative quantification of auxin-related compounds in several A. thaliana genotypes was performed post‐acquisition by normalization to SIL‐IAA in the agarose. SIL-IAA added to agarose was found to remain stable, with repeatability and abundance features comparable to those of other compounds previously used for relative quantification in IR‐ MALDESI analyses.Acetonitrile (ACN)/water and methanol (MeOH)/water solvent gradients from 5 to 95% organic solvent were varied across 2.25 and 4.5 kV applied ESI voltages during IRMALDESI analysis of rat liver tissue. Optimal ESI parameters for lipidomics were determined using longer gradients by the number and abundance of detected lipids and the relative proportion of background ions where positive polarity lipid abundances 3-fold, with 15% greater coverage, while an abundance increase of 1.5-fold and 10% more coverage can be achieved in negative polarity.Kava is an important neuroactive medicinal plant with a large global consumer footprint for its clinical and recreational use, but its production lacks standardization and the tissue-specific metabolite profile of its neuroactive constituents is not well understood. The metabolomic profile of tissues from the roots and stems using an IR-MALDESI MSI approach revealed a unique tissue-specific presence of each target kavalactone. These results provide insights into the use of only peeled roots by linking specific tissue characteristics to concentrations of neuroactive compounds.IR-MALDESI MSI was then used to spatially resolve metabolic profiling of cherry tomatoes. Tomatoes were flash frozen, cryosectioned and imaged with adequate spatial resolution to distinguish between the major tissue structures of a tomato including the skin, mesocarp, endocarp, locular tissue, septum, placenta, seed and seed coating. Metabolites across a diverse array of classes were found, some for the first time in MSI studies, including amino acids, lipids, and several major secondary metabolite classes: terpenes, phenolics, glycosides, and alkaloids. Endogenous carotenoid hydrocarbons, namely lycopene, or its structural isomer β-carotene, were ionized as radical cations.

ISBN: 9798494445728Subjects--Topical Terms:

564049
Physics.

Development of Application Methods of Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (Ir-Maldesi-Msi) of Plants.
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245 1 0 $a Development of Application Methods of Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (Ir-Maldesi-Msi) of Plants.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2021
300 $a 425 p.
500 $a Source: Dissertations Abstracts International, Volume: 83-05, Section: B.
500 $a Advisor: Baker, Erin S. ;Sederoff, Heike Inge;Bowden, Edmond F. ;Muddiman, David C. .
502 $a Thesis (Ph.D.)--North Carolina State University, 2021.
506 $a This item must not be sold to any third party vendors.
520 $a Infrared Matrix Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging (IR-MALDESI-MSI) is a hybrid of matrix assisted laser desorption with electrospray post ionization that enables high resolution spatial information to be combined with the ambient characteristics of electrospray. It has been used to complement other MSI techniques by analyzing samples and biomolecules that are not commonly accessible using the dominant techniques of matrix assisted laser desorption ionization (MALDI) and ESI.The yield fluctuation of artemisinin due to its glandular trichome (GT)-specific biosynthesis has remained an unsolved problem in meeting the global demand for this primary treatment for malaria. Detailed IR-MALDESI image profiling revealed that artemisinin, artemisinic acid, and arteannuin B three metabolites and dihydroartemisinin are localized in non-GT cells of leaves of inbred A. annua plants. Finding that non-GT cells can express the artemisinin biosynthetic pathway led to a revision of the current dogma of artemisinin biosynthesis in A. annua and may expedite innovation of novel metabolic engineering approaches for high and stable production of artemisinin.Quantitative MSI was performed on young Arabidopsis thaliana seedlings to understand how mutations affect metabolite accumulation in plant development. A new method for the quantification of auxin‐related compounds was created using stable‐ isotope‐labelled (SIL) indole‐3‐acetic acid (IAA) doped into agarose substrate. Relative quantification of auxin-related compounds in several A. thaliana genotypes was performed post‐acquisition by normalization to SIL‐IAA in the agarose. SIL-IAA added to agarose was found to remain stable, with repeatability and abundance features comparable to those of other compounds previously used for relative quantification in IR‐ MALDESI analyses.Acetonitrile (ACN)/water and methanol (MeOH)/water solvent gradients from 5 to 95% organic solvent were varied across 2.25 and 4.5 kV applied ESI voltages during IRMALDESI analysis of rat liver tissue. Optimal ESI parameters for lipidomics were determined using longer gradients by the number and abundance of detected lipids and the relative proportion of background ions where positive polarity lipid abundances 3-fold, with 15% greater coverage, while an abundance increase of 1.5-fold and 10% more coverage can be achieved in negative polarity.Kava is an important neuroactive medicinal plant with a large global consumer footprint for its clinical and recreational use, but its production lacks standardization and the tissue-specific metabolite profile of its neuroactive constituents is not well understood. The metabolomic profile of tissues from the roots and stems using an IR-MALDESI MSI approach revealed a unique tissue-specific presence of each target kavalactone. These results provide insights into the use of only peeled roots by linking specific tissue characteristics to concentrations of neuroactive compounds.IR-MALDESI MSI was then used to spatially resolve metabolic profiling of cherry tomatoes. Tomatoes were flash frozen, cryosectioned and imaged with adequate spatial resolution to distinguish between the major tissue structures of a tomato including the skin, mesocarp, endocarp, locular tissue, septum, placenta, seed and seed coating. Metabolites across a diverse array of classes were found, some for the first time in MSI studies, including amino acids, lipids, and several major secondary metabolite classes: terpenes, phenolics, glycosides, and alkaloids. Endogenous carotenoid hydrocarbons, namely lycopene, or its structural isomer β-carotene, were ionized as radical cations.
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