Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes

Research output: Contribution to conferenceConference articleScientific

Abstract

We have designed a novel technology for unravelling the genes involved in the plant secondary metabolism. This technology called Solucel® is based on the genome wide identification and functional analysis of genes involved in the production of phytopharmaceuticals in plant cell cultures. As a model system we used Nicotiana tabacum (BY-2) cell culture in which the nicotine alkaloid production was induced by applying methyl jasmonate as an elicitor (1). Altogether 20 000 transcript tags were visualised. The sequences of the methyl jasmonate modulated (MJM) gene tags were compared to the public databases and 47 of the total 591 MJM gene tags were chosen for further studies. Of particular interest were the genes encoding for protein kinases, signal transducing proteins, transcription factors and other master regulators. Functional analysis of the MJM genes is being performed by using transgenic cell lines of BY-2. Full length cDNAs were constructed and delivered to A. tumefaciens and A. rhizogenes for the establishment of transgenic cell suspension cultures and hairy root cultures, respectively. The transformed cell lines were subjected to metabolite analysis and compared to wild type lines in order to determine the functional properties of the inserted gene construct. In addition, the correlation of the gus-expression and metabolite accumulation in cells carrying promoter-reporter constructs were shown to facilitate the identification of the functional genes. So far, two genes coding for different transcription factors have shown potential to be involved in the regulation of nicotine alkaloid biosynthesis. So-called combinatorial biochemistry approach was applied with the aim of unravelling the possibilities to broaden the chemical diversity of valuable plant derived compounds. The genes derived from tobacco transcriptional profiling were transformed to related or non-related species Hyoscyamus and Catharantus by Agrobacterium transformation, or if required, by using biolistics approaches. Up to date, one gene has shown interesting effect in hairy roots, where besides exceptionally high tropane alkaloid contents, also remarkable changes in secondary metabolite profile was observed, indicating the gene possessing an important role in secondary metabolism. (1) Goossens A, Hakkinen ST, Laakso I, Seppanen-Laakso T, Biondi S, DeSutter V, Lammertyn F, Nuutila AM, Soderlund H, Zabeau M, lnze D, Oksman-Caldentey K-M. Proc Nall Acad Sci 2003; 100: 8595-8600
Original languageEnglish
Publication statusPublished - 2004
MoE publication typeNot Eligible
EventGermany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism - Chiba, Japan
Duration: 20 Sep 200423 Sep 2004

Seminar

SeminarGermany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism
CountryJapan
CityChiba
Period20/09/0423/09/04

Fingerprint

secondary metabolites
metabolism
genes
methyl jasmonate
nicotine
alkaloids
Hyoscyamus
cell culture
transcription factors
cell lines
genetically modified organisms
metabolites
tropane alkaloids
biolistics
Agrobacterium
protein kinases
biochemistry
Nicotiana tabacum
cell suspension culture
phytopharmaceuticals

Cite this

Häkkinen, S. T., Rischer, H., Goossens, A., De Sutter, V., Seppänen-Laakso, T., Ritala, A., ... Oksman-Caldentey, K-M. (2004). Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes. Paper presented at Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism, Chiba, Japan.
Häkkinen, Suvi T. ; Rischer, Heiko ; Goossens, Alain ; De Sutter, Valerie ; Seppänen-Laakso, Tuulikki ; Ritala, Anneli ; Inzé, Dirk ; Oksman-Caldentey, Kirsi-Marja. / Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes. Paper presented at Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism, Chiba, Japan.
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title = "Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes",
abstract = "We have designed a novel technology for unravelling the genes involved in the plant secondary metabolism. This technology called Solucel{\circledR} is based on the genome wide identification and functional analysis of genes involved in the production of phytopharmaceuticals in plant cell cultures. As a model system we used Nicotiana tabacum (BY-2) cell culture in which the nicotine alkaloid production was induced by applying methyl jasmonate as an elicitor (1). Altogether 20 000 transcript tags were visualised. The sequences of the methyl jasmonate modulated (MJM) gene tags were compared to the public databases and 47 of the total 591 MJM gene tags were chosen for further studies. Of particular interest were the genes encoding for protein kinases, signal transducing proteins, transcription factors and other master regulators. Functional analysis of the MJM genes is being performed by using transgenic cell lines of BY-2. Full length cDNAs were constructed and delivered to A. tumefaciens and A. rhizogenes for the establishment of transgenic cell suspension cultures and hairy root cultures, respectively. The transformed cell lines were subjected to metabolite analysis and compared to wild type lines in order to determine the functional properties of the inserted gene construct. In addition, the correlation of the gus-expression and metabolite accumulation in cells carrying promoter-reporter constructs were shown to facilitate the identification of the functional genes. So far, two genes coding for different transcription factors have shown potential to be involved in the regulation of nicotine alkaloid biosynthesis. So-called combinatorial biochemistry approach was applied with the aim of unravelling the possibilities to broaden the chemical diversity of valuable plant derived compounds. The genes derived from tobacco transcriptional profiling were transformed to related or non-related species Hyoscyamus and Catharantus by Agrobacterium transformation, or if required, by using biolistics approaches. Up to date, one gene has shown interesting effect in hairy roots, where besides exceptionally high tropane alkaloid contents, also remarkable changes in secondary metabolite profile was observed, indicating the gene possessing an important role in secondary metabolism. (1) Goossens A, Hakkinen ST, Laakso I, Seppanen-Laakso T, Biondi S, DeSutter V, Lammertyn F, Nuutila AM, Soderlund H, Zabeau M, lnze D, Oksman-Caldentey K-M. Proc Nall Acad Sci 2003; 100: 8595-8600",
author = "H{\"a}kkinen, {Suvi T.} and Heiko Rischer and Alain Goossens and {De Sutter}, Valerie and Tuulikki Sepp{\"a}nen-Laakso and Anneli Ritala and Dirk Inz{\'e} and Kirsi-Marja Oksman-Caldentey",
year = "2004",
language = "English",
note = "Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism ; Conference date: 20-09-2004 Through 23-09-2004",

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Häkkinen, ST, Rischer, H, Goossens, A, De Sutter, V, Seppänen-Laakso, T, Ritala, A, Inzé, D & Oksman-Caldentey, K-M 2004, 'Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes' Paper presented at Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism, Chiba, Japan, 20/09/04 - 23/09/04, .

Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes. / Häkkinen, Suvi T.; Rischer, Heiko; Goossens, Alain; De Sutter, Valerie; Seppänen-Laakso, Tuulikki; Ritala, Anneli; Inzé, Dirk; Oksman-Caldentey, Kirsi-Marja.

2004. Paper presented at Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism, Chiba, Japan.

Research output: Contribution to conferenceConference articleScientific

TY - CONF

T1 - Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes

AU - Häkkinen, Suvi T.

AU - Rischer, Heiko

AU - Goossens, Alain

AU - De Sutter, Valerie

AU - Seppänen-Laakso, Tuulikki

AU - Ritala, Anneli

AU - Inzé, Dirk

AU - Oksman-Caldentey, Kirsi-Marja

PY - 2004

Y1 - 2004

N2 - We have designed a novel technology for unravelling the genes involved in the plant secondary metabolism. This technology called Solucel® is based on the genome wide identification and functional analysis of genes involved in the production of phytopharmaceuticals in plant cell cultures. As a model system we used Nicotiana tabacum (BY-2) cell culture in which the nicotine alkaloid production was induced by applying methyl jasmonate as an elicitor (1). Altogether 20 000 transcript tags were visualised. The sequences of the methyl jasmonate modulated (MJM) gene tags were compared to the public databases and 47 of the total 591 MJM gene tags were chosen for further studies. Of particular interest were the genes encoding for protein kinases, signal transducing proteins, transcription factors and other master regulators. Functional analysis of the MJM genes is being performed by using transgenic cell lines of BY-2. Full length cDNAs were constructed and delivered to A. tumefaciens and A. rhizogenes for the establishment of transgenic cell suspension cultures and hairy root cultures, respectively. The transformed cell lines were subjected to metabolite analysis and compared to wild type lines in order to determine the functional properties of the inserted gene construct. In addition, the correlation of the gus-expression and metabolite accumulation in cells carrying promoter-reporter constructs were shown to facilitate the identification of the functional genes. So far, two genes coding for different transcription factors have shown potential to be involved in the regulation of nicotine alkaloid biosynthesis. So-called combinatorial biochemistry approach was applied with the aim of unravelling the possibilities to broaden the chemical diversity of valuable plant derived compounds. The genes derived from tobacco transcriptional profiling were transformed to related or non-related species Hyoscyamus and Catharantus by Agrobacterium transformation, or if required, by using biolistics approaches. Up to date, one gene has shown interesting effect in hairy roots, where besides exceptionally high tropane alkaloid contents, also remarkable changes in secondary metabolite profile was observed, indicating the gene possessing an important role in secondary metabolism. (1) Goossens A, Hakkinen ST, Laakso I, Seppanen-Laakso T, Biondi S, DeSutter V, Lammertyn F, Nuutila AM, Soderlund H, Zabeau M, lnze D, Oksman-Caldentey K-M. Proc Nall Acad Sci 2003; 100: 8595-8600

AB - We have designed a novel technology for unravelling the genes involved in the plant secondary metabolism. This technology called Solucel® is based on the genome wide identification and functional analysis of genes involved in the production of phytopharmaceuticals in plant cell cultures. As a model system we used Nicotiana tabacum (BY-2) cell culture in which the nicotine alkaloid production was induced by applying methyl jasmonate as an elicitor (1). Altogether 20 000 transcript tags were visualised. The sequences of the methyl jasmonate modulated (MJM) gene tags were compared to the public databases and 47 of the total 591 MJM gene tags were chosen for further studies. Of particular interest were the genes encoding for protein kinases, signal transducing proteins, transcription factors and other master regulators. Functional analysis of the MJM genes is being performed by using transgenic cell lines of BY-2. Full length cDNAs were constructed and delivered to A. tumefaciens and A. rhizogenes for the establishment of transgenic cell suspension cultures and hairy root cultures, respectively. The transformed cell lines were subjected to metabolite analysis and compared to wild type lines in order to determine the functional properties of the inserted gene construct. In addition, the correlation of the gus-expression and metabolite accumulation in cells carrying promoter-reporter constructs were shown to facilitate the identification of the functional genes. So far, two genes coding for different transcription factors have shown potential to be involved in the regulation of nicotine alkaloid biosynthesis. So-called combinatorial biochemistry approach was applied with the aim of unravelling the possibilities to broaden the chemical diversity of valuable plant derived compounds. The genes derived from tobacco transcriptional profiling were transformed to related or non-related species Hyoscyamus and Catharantus by Agrobacterium transformation, or if required, by using biolistics approaches. Up to date, one gene has shown interesting effect in hairy roots, where besides exceptionally high tropane alkaloid contents, also remarkable changes in secondary metabolite profile was observed, indicating the gene possessing an important role in secondary metabolism. (1) Goossens A, Hakkinen ST, Laakso I, Seppanen-Laakso T, Biondi S, DeSutter V, Lammertyn F, Nuutila AM, Soderlund H, Zabeau M, lnze D, Oksman-Caldentey K-M. Proc Nall Acad Sci 2003; 100: 8595-8600

M3 - Conference article

ER -

Häkkinen ST, Rischer H, Goossens A, De Sutter V, Seppänen-Laakso T, Ritala A et al. Discovering plant secondary metabolite pathways – functional analysis of secondary metabolism related genes. 2004. Paper presented at Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism, Chiba, Japan.