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Title: Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging

Abstract

Reverse genetic strategies, such as virus-induced gene silencing, are powerful techniques to study gene function. Currently, there are few tools to study the spatial dependence of the consequences of gene silencing at the cellular level. Here, we report the use of multimodal Raman and mass spectrometry imaging to study the cellular-level biochemical changes that occur from silencing the phytoene desaturase ( pds) gene using a Foxtail mosaic virus (FoMV) vector in maize leaves. The multimodal imaging method allows the localized carotenoid distribution to be measured and reveals differences lost in the spatial average when analyzing a carotenoid extraction of the whole leaf. The nature of the Raman and mass spectrometry signals are complementary: silencing pds reduces the downstream carotenoid Raman signal and increases the phytoene mass spectrometry signal.

Authors:
 [1];  [1];  [2];  [1];  [1];  [2];  [2];  [1];  [2];  [2]; ORCiD logo [1]
  1. Ames Lab. and Iowa State Univ., Ames, IA (United States)
  2. Iowa State Univ., Ames, IA (United States)
Publication Date:
Research Org.:
Ames Laboratory (AMES), Ames, IA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1440994
Report Number(s):
IS-J-9659
Journal ID: ISSN 1746-4811; PII: 306
Grant/Contract Number:  
AC02-07CH11358; SC0014038; Hatch Project No. 3808
Resource Type:
Accepted Manuscript
Journal Name:
Plant Methods
Additional Journal Information:
Journal Volume: 14; Journal Issue: 1; Journal ID: ISSN 1746-4811
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; RNA silencing; Foxtail mosaic virus; Phytoene desaturase; Subcellular Raman imaging; Mass spectrometry imaging; Whole-plant analysis; Biochemical characterization; Mosaic spatial pattern; Carotenoids

Citation Formats

Burkhow, Sadie J., Stephens, Nicole M., Mei, Yu, Duenas, Maria Emilia, Freppon, Daniel J., Ding, Geng, Smith, Shea C., Lee, Young -Jin, Nikolau, Basil J., Whitham, Steven A., and Smith, Emily A. Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging. United States: N. p., 2018. Web. doi:10.1186/s13007-018-0306-7.
Burkhow, Sadie J., Stephens, Nicole M., Mei, Yu, Duenas, Maria Emilia, Freppon, Daniel J., Ding, Geng, Smith, Shea C., Lee, Young -Jin, Nikolau, Basil J., Whitham, Steven A., & Smith, Emily A. Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging. United States. doi:10.1186/s13007-018-0306-7.
Burkhow, Sadie J., Stephens, Nicole M., Mei, Yu, Duenas, Maria Emilia, Freppon, Daniel J., Ding, Geng, Smith, Shea C., Lee, Young -Jin, Nikolau, Basil J., Whitham, Steven A., and Smith, Emily A. Fri . "Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging". United States. doi:10.1186/s13007-018-0306-7. https://www.osti.gov/servlets/purl/1440994.
@article{osti_1440994,
title = {Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging},
author = {Burkhow, Sadie J. and Stephens, Nicole M. and Mei, Yu and Duenas, Maria Emilia and Freppon, Daniel J. and Ding, Geng and Smith, Shea C. and Lee, Young -Jin and Nikolau, Basil J. and Whitham, Steven A. and Smith, Emily A.},
abstractNote = {Reverse genetic strategies, such as virus-induced gene silencing, are powerful techniques to study gene function. Currently, there are few tools to study the spatial dependence of the consequences of gene silencing at the cellular level. Here, we report the use of multimodal Raman and mass spectrometry imaging to study the cellular-level biochemical changes that occur from silencing the phytoene desaturase (pds) gene using a Foxtail mosaic virus (FoMV) vector in maize leaves. The multimodal imaging method allows the localized carotenoid distribution to be measured and reveals differences lost in the spatial average when analyzing a carotenoid extraction of the whole leaf. The nature of the Raman and mass spectrometry signals are complementary: silencing pds reduces the downstream carotenoid Raman signal and increases the phytoene mass spectrometry signal.},
doi = {10.1186/s13007-018-0306-7},
journal = {Plant Methods},
number = 1,
volume = 14,
place = {United States},
year = {2018},
month = {5}
}

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