Please use this identifier to cite or link to this item:
http://hdl.handle.net/2445/121632
Title: | Nerolidol production in agroinfiltrated tobacco: Impact of protein stability and membrane targeting of strawberry (Fragraria ananassa) NEROLIDOL SYNTHASE1 |
Author: | Andrade Poveda, Paola Andrea Manzano Alías, David Ramírez Estrada, Karla Caudepón, Daniel Arró i Plans, Montserrat Ferrer i Prats, Albert Phillips, Michael A. |
Keywords: | Biotecnologia agrícola Extracció (Química) Terpens Olis essencials Tabac Agricultural biotechnology Extraction (Chemistry) Terpenes Essences and essential oils Tobacco |
Issue Date: | Feb-2018 |
Publisher: | Elsevier B.V. |
Abstract: | The sesquiterpene alcohol nerolidol, synthesized from farnesyl diphosphate (FDP), mediates plant-insect inter- actions across multiple trophic levels with major implications for pest management in agriculture. We compared nerolidol engineering strategies in tobacco using agroinfiltration to transiently express strawberry (Fragraria ananassa) linalool/nerolidol synthase (FaNES1) either at the endoplasmic reticulum (ER) or in the cytosol as a soluble protein. Using solid phase microextraction and gas chromatography-mass spectrometry (SPME-GCMS), we have determined that FaNES1 directed to the ER via fusion to the transmembrane domain of squalene synthase or hydroxymethylglutaryl - CoA reductase displayed significant improvements in terms of transcript levels, protein accumulation, and volatile production when compared to its cytosolic form. However, the highest levels of nerolidol production were observed when FaNES1 was fused to GFP and expressed in the cytosol. This SPME-GCMS method afforded a limit of detection and quantification of 1.54 and 5.13 pg, respectively. Nerolidol production levels, which ranged from 0.5 to 3.0 μg/g F.W., correlated more strongly to the accumulation of recombinant protein than transcript level, the former being highest in FaNES-GFP transfected plants. These results indicate that while the ER may represent an enriched source of FDP that can be exploited in metabolic engineering, protein accumulation is a better predictor of sesquiterpene production. |
Note: | Versió postprint del document publicat a: https://doi.org/10.1016/j.plantsci.2017.11.013 |
It is part of: | Plant Science, 2017, vol. 267, p. 112-123 |
URI: | http://hdl.handle.net/2445/121632 |
Related resource: | https://doi.org/10.1016/j.plantsci.2017.11.013 |
ISSN: | 0168-9452 |
Appears in Collections: | Articles publicats en revistes (Bioquímica i Fisiologia) |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
674740.pdf | 6.04 MB | Adobe PDF | View/Open |
This item is licensed under a
Creative Commons License