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Article
Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging
Plant Methods
  • Sadie J. Burkhow, Iowa State University and Ames Laboratory
  • Nicole M. Stephens, Iowa State University and Ames Laboratory
  • Yu Mei, Iowa State University
  • Maria Emilia Duenas, Iowa State University and Ames Laboratory
  • Daniel J. Freppon, Iowa State University and Ames Laboratory
  • Geng Ding, Iowa State University
  • Shea C. Smith, Iowa State University
  • Young-Jin Lee, Iowa State University and Ames Laboratory
  • Basil J. Nikolau, Iowa State University
  • Steven A. Whitham, Iowa State University
  • Emily A. Smith, Iowa State University and Ames Laboratory
Document Type
Article
Publication Version
Published Version
Publication Date
5-25-2018
DOI
10.1186/s13007-018-0306-7
Abstract

Background: 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.

Results: 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.

Conclusions: Both Raman and mass spectrometry imaging show that the biochemical changes from FoMV-pds silencing occur with a mosaic spatial pattern at the cellular level, and the Raman images show carotenoid expression was reduced at discrete locations but not eliminated. The data indicate the multimodal imaging method has great utility to study the biochemical changes that result from gene silencing at the cellular spatial level of expression in many plant tissues including the stem and leaf. Our demonstrated method is the first to spatially characterize the biochemical changes as a result of VIGS at the cellular level using commonly available instrumentation.

Comments

This article is published as Burkhow, Sadie J., Nicole M. Stephens, Yu Mei, Maria Emilia Dueñas, Daniel J. Freppon, Geng Ding, Shea C. Smith et al. "Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging." Plant Methods 14, no. 1 (2018): 37.

Creative Commons License
Creative Commons Attribution 4.0
Copyright Owner
The Authors
Language
en
File Format
application/pdf
Citation Information
Sadie J. Burkhow, Nicole M. Stephens, Yu Mei, Maria Emilia Duenas, et al.. "Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging" Plant Methods Vol. 14 Iss. 1 (2018) p. 37
Available at: http://works.bepress.com/emily-smith/32/