Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves

Thierry L. Delatte, Giulia Scaiola, Jamil Molenaar, Katyuce de Sousa Farias, Leticia Alves Gomes Albertti, Jacqueline Busscher, Francel Verstappen, Carlos Carollo, Harro Bouwmeester, Jules Beekwilder

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)

Abstract

Plants store volatile compounds in specialized organs. The properties of these storage organs prevent precarious evaporation and protect neighbouring tissues from cytotoxicity. Metabolic engineering of plants is often carried out in tissues such as leaf mesophyll cells, which are abundant and easily accessible by engineering tools. However, these tissues are not suitable for the storage of volatile and hydrophobic compound such as sesquiterpenes and engineered volatiles are often lost into the headspace. In this study, we show that the seeds of Arabidopsis thaliana, which naturally contain lipid bodies, accumulate sesquiterpenes upon engineered expression. Subsequently, storage of volatile sesquiterpenes was achieved in Nicotiana benthamiana leaf tissue, by introducing oleosin-coated lipid bodies through metabolic engineering. Hereto, different combinations of genes encoding diacylglycerol acyltransferases (DGATs), transcription factors (WRINKL1) and oleosins (OLE1), from the oil seed-producing species castor bean (Ricinus communis) and Arabidopsis, were assessed for their suitability to promote lipid body formation. Co-expression of α-bisabolol synthase with Arabidopsis DGAT1 and WRINKL1 and OLE1 from castor bean promoted storage of α-bisabolol in N. benthamiana mesophyll tissue more than 17-fold. A clear correlation was found between neutral lipids and storage of sesquiterpenes, using synthases for α-bisabolol, (E)-β-caryophyllene and α-barbatene. The co-localization of neutral lipids and α-bisabolol was shown using microscopy. This work demonstrates that lipid bodies can be used as intracellular storage compartment for hydrophobic sesquiterpenes, also in the vegetative parts of plants, creating the possibility to improve yields of metabolic engineering strategies in plants.

LanguageEnglish
Pages1997-2006
JournalPlant Biotechnology Journal
Volume16
Issue number12
Early online date23 Apr 2018
DOIs
Publication statusPublished - Dec 2018

Fingerprint

alpha-bisabolol
Nicotiana benthamiana
Sesquiterpenes
sesquiterpenoids
lipid bodies
Tobacco
engineering
Metabolic Engineering
metabolic engineering
Castor Bean
Arabidopsis
oleosin
castor beans
leaves
mesophyll
Seeds
Diacylglycerol O-Acyltransferase
triacylglycerols
Mesophyll Cells
diacylglycerol acyltransferase

Keywords

  • Nicotiana benthamiana
  • Lipid bodies
  • Secondary metabolites
  • Sesquiterpene
  • Storage
  • Triacylglycerol

Cite this

Delatte, Thierry L. ; Scaiola, Giulia ; Molenaar, Jamil ; de Sousa Farias, Katyuce ; Alves Gomes Albertti, Leticia ; Busscher, Jacqueline ; Verstappen, Francel ; Carollo, Carlos ; Bouwmeester, Harro ; Beekwilder, Jules. / Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves. In: Plant Biotechnology Journal. 2018 ; Vol. 16, No. 12. pp. 1997-2006.
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title = "Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves",
abstract = "Plants store volatile compounds in specialized organs. The properties of these storage organs prevent precarious evaporation and protect neighbouring tissues from cytotoxicity. Metabolic engineering of plants is often carried out in tissues such as leaf mesophyll cells, which are abundant and easily accessible by engineering tools. However, these tissues are not suitable for the storage of volatile and hydrophobic compound such as sesquiterpenes and engineered volatiles are often lost into the headspace. In this study, we show that the seeds of Arabidopsis thaliana, which naturally contain lipid bodies, accumulate sesquiterpenes upon engineered expression. Subsequently, storage of volatile sesquiterpenes was achieved in Nicotiana benthamiana leaf tissue, by introducing oleosin-coated lipid bodies through metabolic engineering. Hereto, different combinations of genes encoding diacylglycerol acyltransferases (DGATs), transcription factors (WRINKL1) and oleosins (OLE1), from the oil seed-producing species castor bean (Ricinus communis) and Arabidopsis, were assessed for their suitability to promote lipid body formation. Co-expression of α-bisabolol synthase with Arabidopsis DGAT1 and WRINKL1 and OLE1 from castor bean promoted storage of α-bisabolol in N. benthamiana mesophyll tissue more than 17-fold. A clear correlation was found between neutral lipids and storage of sesquiterpenes, using synthases for α-bisabolol, (E)-β-caryophyllene and α-barbatene. The co-localization of neutral lipids and α-bisabolol was shown using microscopy. This work demonstrates that lipid bodies can be used as intracellular storage compartment for hydrophobic sesquiterpenes, also in the vegetative parts of plants, creating the possibility to improve yields of metabolic engineering strategies in plants.",
keywords = "Nicotiana benthamiana, Lipid bodies, Secondary metabolites, Sesquiterpene, Storage, Triacylglycerol",
author = "Delatte, {Thierry L.} and Giulia Scaiola and Jamil Molenaar and {de Sousa Farias}, Katyuce and {Alves Gomes Albertti}, Leticia and Jacqueline Busscher and Francel Verstappen and Carlos Carollo and Harro Bouwmeester and Jules Beekwilder",
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Delatte, TL, Scaiola, G, Molenaar, J, de Sousa Farias, K, Alves Gomes Albertti, L, Busscher, J, Verstappen, F, Carollo, C, Bouwmeester, H & Beekwilder, J 2018, 'Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves', Plant Biotechnology Journal, vol. 16, no. 12, pp. 1997-2006. https://doi.org/10.1111/pbi.12933

Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves. / Delatte, Thierry L.; Scaiola, Giulia; Molenaar, Jamil; de Sousa Farias, Katyuce; Alves Gomes Albertti, Leticia; Busscher, Jacqueline; Verstappen, Francel; Carollo, Carlos; Bouwmeester, Harro; Beekwilder, Jules.

In: Plant Biotechnology Journal, Vol. 16, No. 12, 12.2018, p. 1997-2006.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Delatte, Thierry L.

AU - Scaiola, Giulia

AU - Molenaar, Jamil

AU - de Sousa Farias, Katyuce

AU - Alves Gomes Albertti, Leticia

AU - Busscher, Jacqueline

AU - Verstappen, Francel

AU - Carollo, Carlos

AU - Bouwmeester, Harro

AU - Beekwilder, Jules

PY - 2018/12

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AB - Plants store volatile compounds in specialized organs. The properties of these storage organs prevent precarious evaporation and protect neighbouring tissues from cytotoxicity. Metabolic engineering of plants is often carried out in tissues such as leaf mesophyll cells, which are abundant and easily accessible by engineering tools. However, these tissues are not suitable for the storage of volatile and hydrophobic compound such as sesquiterpenes and engineered volatiles are often lost into the headspace. In this study, we show that the seeds of Arabidopsis thaliana, which naturally contain lipid bodies, accumulate sesquiterpenes upon engineered expression. Subsequently, storage of volatile sesquiterpenes was achieved in Nicotiana benthamiana leaf tissue, by introducing oleosin-coated lipid bodies through metabolic engineering. Hereto, different combinations of genes encoding diacylglycerol acyltransferases (DGATs), transcription factors (WRINKL1) and oleosins (OLE1), from the oil seed-producing species castor bean (Ricinus communis) and Arabidopsis, were assessed for their suitability to promote lipid body formation. Co-expression of α-bisabolol synthase with Arabidopsis DGAT1 and WRINKL1 and OLE1 from castor bean promoted storage of α-bisabolol in N. benthamiana mesophyll tissue more than 17-fold. A clear correlation was found between neutral lipids and storage of sesquiterpenes, using synthases for α-bisabolol, (E)-β-caryophyllene and α-barbatene. The co-localization of neutral lipids and α-bisabolol was shown using microscopy. This work demonstrates that lipid bodies can be used as intracellular storage compartment for hydrophobic sesquiterpenes, also in the vegetative parts of plants, creating the possibility to improve yields of metabolic engineering strategies in plants.

KW - Nicotiana benthamiana

KW - Lipid bodies

KW - Secondary metabolites

KW - Sesquiterpene

KW - Storage

KW - Triacylglycerol

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T2 - Plant Biotechnology Journal

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Delatte TL, Scaiola G, Molenaar J, de Sousa Farias K, Alves Gomes Albertti L, Busscher J et al. Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves. Plant Biotechnology Journal. 2018 Dec;16(12):1997-2006. https://doi.org/10.1111/pbi.12933