PMID- 29849743 OWN - NLM STAT- PubMed-not-MEDLINE VI - 14 TI - Characterizing virus-induced gene silencing at the cellular level with in situ multimodal imaging. PG - 37 LA - eng PT - Journal Article PL - England TA - Plant Methods JT - Plant methods JID - 101245798 IS - 1746-4811 (Print) LID - 10.1186/s13007-018-0306-7 [doi] FAU - Burkhow, Sadie J AU - Burkhow SJ AD - 1The Ames Laboratory, U.S. Department of Energy, Iowa State University, Ames, IA 50011-3111 USA. AD - 2Department of Chemistry, Iowa State University, Ames, IA 50011-3111 USA. FAU - Stephens, Nicole M AU - Stephens NM AD - 1The Ames Laboratory, U.S. Department of Energy, Iowa State University, Ames, IA 50011-3111 USA. AD - 2Department of Chemistry, Iowa State University, Ames, IA 50011-3111 USA. FAU - Mei, Yu AU - Mei Y AD - 3Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA 50011 USA. FAU - Dueñas, Maria Emilia AU - Dueñas ME AD - 1The Ames Laboratory, U.S. Department of Energy, Iowa State University, Ames, IA 50011-3111 USA. AD - 2Department of Chemistry, Iowa State University, Ames, IA 50011-3111 USA. FAU - Freppon, Daniel J AU - Freppon DJ AD - 1The Ames Laboratory, U.S. Department of Energy, Iowa State University, Ames, IA 50011-3111 USA. AD - 2Department of Chemistry, Iowa State University, Ames, IA 50011-3111 USA. FAU - Ding, Geng AU - Ding G AD - 4Department of Biochemistry Biophysics, and Molecular Biology, Center for Metabolic Biology, Iowa State University, Ames, IA 50011 USA. FAU - Smith, Shea C AU - Smith SC AD - 5Engineering Research Center for Biorenewable Chemicals, Iowa State University, Ames, IA 50011 USA. FAU - Lee, Young-Jin AU - Lee YJ AD - 1The Ames Laboratory, U.S. Department of Energy, Iowa State University, Ames, IA 50011-3111 USA. AD - 2Department of Chemistry, Iowa State University, Ames, IA 50011-3111 USA. FAU - Nikolau, Basil J AU - Nikolau BJ AD - 4Department of Biochemistry Biophysics, and Molecular Biology, Center for Metabolic Biology, Iowa State University, Ames, IA 50011 USA. FAU - Whitham, Steven A AU - Whitham SA AD - 3Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA 50011 USA. FAU - Smith, Emily A AU - Smith EA AUID- ORCID: 0000-0001-7438-7808 AD - 1The Ames Laboratory, U.S. Department of Energy, Iowa State University, Ames, IA 50011-3111 USA. AD - 2Department of Chemistry, Iowa State University, Ames, IA 50011-3111 USA. IS - 1746-4811 (Linking) OTO - NOTNLM OT - Biochemical characterization OT - Carotenoids OT - Foxtail mosaic virus OT - Mass spectrometry imaging OT - Mosaic spatial pattern OT - Phytoene desaturase OT - RNA silencing OT - Subcellular Raman imaging OT - Whole-plant analysis PMC - PMC5968576 LR - 20201001 DP - 2018 DEP - 20180525 AB - 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.