Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis

Suvi T. Häkkinen, Heiko Rischer, Alain Goossens, Valerie De Sutter, Dirk Inzé, Kirsi-Marja Oksman-Caldentey

Research output: Contribution to conferenceConference articleScientific

Abstract

Genetic knowledge on biosynthetic pathways and the regulation thereof is of crucial importance to bypass the low-product yield of various secondary metabolites in plant cells. To facilitate gene discovery in plant secondary metabolism, we have developed a comprehensive profiling approach that is based on functional genomics and can be applied on any plant system without a need for prior sequence knowledge. Jasmonate-induced changes have been monitored on the transcript and alkaloid profiles of tobacco. Using cDNA-AFLP based transcript profiling, an inventory of hundreds of genes, potentially involved in plant secondary metabolism, has been built. A technology platform, especially driven towards the exploitation of plant cell cultures, for high-throughput isolation and functional analysis of these genes, has been established. At present, more than 50 tobacco BY-2 full-length open reading frames have been isolated and are currently subjected to functional analysis. In our tobacco BY-2 model system the overexpression of 22 genes resulted in three lines exhibiting an altered alkaloid accumulation pattern compared to controls. The genes derived from the tobacco transcriptional profiling are also used for the transformation of other plant species. So far, one of the tested genes has strongly influenced the tropane alkaloid biosynthesis, which is partly the same as for nicotine alkaloids. We have developed a technology platform called SOLUCEL that allows creating a novel toolbox for metabolic engineering of plant cells. It can be applied (i) to enhance the production of marketed high-value pharmaceuticals in plant cell cultures (ii) to develop reliable and reproducible sources of plant-derived molecules with potential pharmaceutical value, and (iii) to increase the chemical diversity of plant based molecules through Combinatorial Biochemistry.
Original languageEnglish
Publication statusPublished - 2004
MoE publication typeNot Eligible
EventInternational Symposium Cell and Molecular Biology of Tobacco BY-2 Cells - Yokohama, Japan
Duration: 14 Sep 200416 Sep 2004

Conference

ConferenceInternational Symposium Cell and Molecular Biology of Tobacco BY-2 Cells
CountryJapan
CityYokohama
Period14/09/0416/09/04

Fingerprint

metabolic engineering
tobacco
metabolites
metabolism
alkaloids
genes
cell culture
tropane alkaloids
drugs
nicotine
biochemistry
secondary metabolites
amplified fragment length polymorphism
open reading frames
biochemical pathways
biosynthesis
cells
genomics

Cite this

Häkkinen, S. T., Rischer, H., Goossens, A., De Sutter, V., Inzé, D., & Oksman-Caldentey, K-M. (2004). Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis. Paper presented at International Symposium Cell and Molecular Biology of Tobacco BY-2 Cells , Yokohama, Japan.
Häkkinen, Suvi T. ; Rischer, Heiko ; Goossens, Alain ; De Sutter, Valerie ; Inzé, Dirk ; Oksman-Caldentey, Kirsi-Marja. / Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis. Paper presented at International Symposium Cell and Molecular Biology of Tobacco BY-2 Cells , Yokohama, Japan.
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abstract = "Genetic knowledge on biosynthetic pathways and the regulation thereof is of crucial importance to bypass the low-product yield of various secondary metabolites in plant cells. To facilitate gene discovery in plant secondary metabolism, we have developed a comprehensive profiling approach that is based on functional genomics and can be applied on any plant system without a need for prior sequence knowledge. Jasmonate-induced changes have been monitored on the transcript and alkaloid profiles of tobacco. Using cDNA-AFLP based transcript profiling, an inventory of hundreds of genes, potentially involved in plant secondary metabolism, has been built. A technology platform, especially driven towards the exploitation of plant cell cultures, for high-throughput isolation and functional analysis of these genes, has been established. At present, more than 50 tobacco BY-2 full-length open reading frames have been isolated and are currently subjected to functional analysis. In our tobacco BY-2 model system the overexpression of 22 genes resulted in three lines exhibiting an altered alkaloid accumulation pattern compared to controls. The genes derived from the tobacco transcriptional profiling are also used for the transformation of other plant species. So far, one of the tested genes has strongly influenced the tropane alkaloid biosynthesis, which is partly the same as for nicotine alkaloids. We have developed a technology platform called SOLUCEL that allows creating a novel toolbox for metabolic engineering of plant cells. It can be applied (i) to enhance the production of marketed high-value pharmaceuticals in plant cell cultures (ii) to develop reliable and reproducible sources of plant-derived molecules with potential pharmaceutical value, and (iii) to increase the chemical diversity of plant based molecules through Combinatorial Biochemistry.",
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Häkkinen, ST, Rischer, H, Goossens, A, De Sutter, V, Inzé, D & Oksman-Caldentey, K-M 2004, 'Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis' Paper presented at International Symposium Cell and Molecular Biology of Tobacco BY-2 Cells , Yokohama, Japan, 14/09/04 - 16/09/04, .

Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis. / Häkkinen, Suvi T.; Rischer, Heiko; Goossens, Alain; De Sutter, Valerie; Inzé, Dirk; Oksman-Caldentey, Kirsi-Marja.

2004. Paper presented at International Symposium Cell and Molecular Biology of Tobacco BY-2 Cells , Yokohama, Japan.

Research output: Contribution to conferenceConference articleScientific

TY - CONF

T1 - Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis

AU - Häkkinen, Suvi T.

AU - Rischer, Heiko

AU - Goossens, Alain

AU - De Sutter, Valerie

AU - Inzé, Dirk

AU - Oksman-Caldentey, Kirsi-Marja

PY - 2004

Y1 - 2004

N2 - Genetic knowledge on biosynthetic pathways and the regulation thereof is of crucial importance to bypass the low-product yield of various secondary metabolites in plant cells. To facilitate gene discovery in plant secondary metabolism, we have developed a comprehensive profiling approach that is based on functional genomics and can be applied on any plant system without a need for prior sequence knowledge. Jasmonate-induced changes have been monitored on the transcript and alkaloid profiles of tobacco. Using cDNA-AFLP based transcript profiling, an inventory of hundreds of genes, potentially involved in plant secondary metabolism, has been built. A technology platform, especially driven towards the exploitation of plant cell cultures, for high-throughput isolation and functional analysis of these genes, has been established. At present, more than 50 tobacco BY-2 full-length open reading frames have been isolated and are currently subjected to functional analysis. In our tobacco BY-2 model system the overexpression of 22 genes resulted in three lines exhibiting an altered alkaloid accumulation pattern compared to controls. The genes derived from the tobacco transcriptional profiling are also used for the transformation of other plant species. So far, one of the tested genes has strongly influenced the tropane alkaloid biosynthesis, which is partly the same as for nicotine alkaloids. We have developed a technology platform called SOLUCEL that allows creating a novel toolbox for metabolic engineering of plant cells. It can be applied (i) to enhance the production of marketed high-value pharmaceuticals in plant cell cultures (ii) to develop reliable and reproducible sources of plant-derived molecules with potential pharmaceutical value, and (iii) to increase the chemical diversity of plant based molecules through Combinatorial Biochemistry.

AB - Genetic knowledge on biosynthetic pathways and the regulation thereof is of crucial importance to bypass the low-product yield of various secondary metabolites in plant cells. To facilitate gene discovery in plant secondary metabolism, we have developed a comprehensive profiling approach that is based on functional genomics and can be applied on any plant system without a need for prior sequence knowledge. Jasmonate-induced changes have been monitored on the transcript and alkaloid profiles of tobacco. Using cDNA-AFLP based transcript profiling, an inventory of hundreds of genes, potentially involved in plant secondary metabolism, has been built. A technology platform, especially driven towards the exploitation of plant cell cultures, for high-throughput isolation and functional analysis of these genes, has been established. At present, more than 50 tobacco BY-2 full-length open reading frames have been isolated and are currently subjected to functional analysis. In our tobacco BY-2 model system the overexpression of 22 genes resulted in three lines exhibiting an altered alkaloid accumulation pattern compared to controls. The genes derived from the tobacco transcriptional profiling are also used for the transformation of other plant species. So far, one of the tested genes has strongly influenced the tropane alkaloid biosynthesis, which is partly the same as for nicotine alkaloids. We have developed a technology platform called SOLUCEL that allows creating a novel toolbox for metabolic engineering of plant cells. It can be applied (i) to enhance the production of marketed high-value pharmaceuticals in plant cell cultures (ii) to develop reliable and reproducible sources of plant-derived molecules with potential pharmaceutical value, and (iii) to increase the chemical diversity of plant based molecules through Combinatorial Biochemistry.

M3 - Conference article

ER -

Häkkinen ST, Rischer H, Goossens A, De Sutter V, Inzé D, Oksman-Caldentey K-M. Metabolic engineering of tobacco BY-2 secondary metabolism based on transcriptional profiling and metabolite analysis. 2004. Paper presented at International Symposium Cell and Molecular Biology of Tobacco BY-2 Cells , Yokohama, Japan.