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 language | English |
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Publication status | Published - 2004 |
MoE publication type | Not Eligible |
Event | Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism - Chiba, Japan Duration: 20 Sept 2004 → 23 Sept 2004 |
Seminar
Seminar | Germany – Japan Seminar on Molecular Regulation of Plant Secondary Metabolism |
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Country/Territory | Japan |
City | Chiba |
Period | 20/09/04 → 23/09/04 |