Abstract
The aim of this study was to discover genes involved in grape secondary metabolism pathways by stimulating their production in grape cell suspension cultures. Among numerous elicitors tested, methyl jasmonate (MeJA) induced the production of a wide range of sesquiterpenes in Cabernet Sauvignon cell suspension cultures derived from berries. The efficacy of MeJA in the induction of sesquiterpenes production is strictly dependent upon the density of cells in cell suspension at the moment of elicitor addition. We also found that jasmonic acid (JA) as MeJA activate sesquiterpene synthesis, but when salicylic acid (SA) was added in addition to MeJA, sesquiterpenes were not produced. Microarray analyses on the induced cultures confirmed the activation of sesquiterpenes biosynthetic pathway by MeJA and JA and the inhibition effect of SA: two terpene synthases on the gene chip were upregulated in the MeJA and JA treated cells. Microarray data also indicated that genes from tannins and stilbenes pathways are differentially expressed in response to the treatments. Tannins accumulated to higher levels in the MeJA- and JA-treated cell suspension cultures and the expressions of pathway key genes was more than 2-fold higher in the MeJA and JA treatments than the control and MeJA+SA cells. Stilbenes accumulated in all three treatments, but the levels correlated mostly with the expression of stilbene synthase (STS2). The results of this work confirm that an inducible cell culture system is a powerful tool for functional genomics studies as we can associate specific genes from biosynthesis pathways with changes in metabolite production. We are now working towards the functional characterisation of terpene synthases, modifying enzymes and transport proteins associated with volatile production in grapes. We have also identified several transcription factors that have expression patterns matching the induction patterns seen for the three pathways described above. These are also targets of future work as they may help us understand the regulation of these secondary metabolite pathways in grape berries.
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Claims 14
Claim · model-written summary
Methyl jasmonate induced sesquiterpene production in Cabernet Sauvignon grape cell cultures.
- Taxon
- grapet0
- Material
- Cabernet Sauvignone0
- Organ / stage
- berriese0
- Treatment
- methyl jasmonate (MeJA)e0
- Outcome
- production of a wide range of sesquiterpenese0
- Direction
- inducede0
Not reportedComparator
Verbatim
e0“methyl jasmonate (MeJA) induced the production of a wide range of sesquiterpenes in Cabernet Sauvignon cell suspension cultures derived from berries”
t0“grape”
Claim · model-written summary
MeJA induction of sesquiterpenes depended on cell density at addition.
- Taxon
- grapet0
- Treatment
- MeJAe0
- Outcome
- induction of sesquiterpenes productione0
- Qualifier
- density of cells in cell suspension at the moment of elicitor additione0
Not reportedDirection
Verbatim
e0“The efficacy of MeJA in the induction of sesquiterpenes production is strictly dependent upon the density of cells in cell suspension at the moment of elicitor addition”
t0“grape”
Claim · model-written summary
Jasmonic acid activated sesquiterpene synthesis.
Not reportedComparator
Verbatim
e0“jasmonic acid (JA) as MeJA activate sesquiterpene synthesis”
t0“grape”
Claim · model-written summary
Adding salicylic acid to MeJA prevented sesquiterpene production.
- Taxon
- grapet0
- Treatment
- salicylic acid (SA) was added in addition to MeJAe0
- Outcome
- sesquiterpenes were not producede0
- Comparator
- MeJAe0
Not reportedDirection
Verbatim
e0“when salicylic acid (SA) was added in addition to MeJA, sesquiterpenes were not produced”
t0“grape”
Claim · model-written summary
Two terpene synthases were upregulated after MeJA treatment.
- Taxon
- grapet0
- Treatment
- MeJAe0
- Gene
- terpene synthasese0
- Outcome
- terpene synthases on the gene chipe0
- Direction
- upregulatede0
Not reportedComparator
Verbatim
e0“two terpene synthases on the gene chip were upregulated in the MeJA and JA treated cells”
t0“grape”
Claim · model-written summary
Two terpene synthases were upregulated after JA treatment.
- Taxon
- grapet0
- Treatment
- JAe0
- Gene
- terpene synthasese0
- Outcome
- terpene synthases on the gene chipe0
- Direction
- upregulatede0
Not reportedComparator
Verbatim
e0“two terpene synthases on the gene chip were upregulated in the MeJA and JA treated cells”
t0“grape”
Claim · model-written summary
Tannin and stilbene pathway genes responded differentially to treatments.
Verbatim
e0“genes from tannins and stilbenes pathways are differentially expressed in response to the treatments”
t0“grape”
Claim · model-written summary
Tannins accumulated to higher levels in MeJA treated cells.
Not reportedComparator
Verbatim
e0“Tannins accumulated to higher levels in the MeJA- and JA-treated cell suspension cultures”
t0“grape”
Claim · model-written summary
Tannins accumulated to higher levels in JA treated cells.
Not reportedComparator
Verbatim
e0“Tannins accumulated to higher levels in the MeJA- and JA-treated cell suspension cultures”
t0“grape”
Claim · model-written summary
Tannin pathway key gene expression was over twofold higher with MeJA than control and MeJA plus SA.
Verbatim
e0“expressions of pathway key genes was more than 2-fold higher in the MeJA and JA treatments than the control and MeJA+SA cells”
t0“grape”
Claim · model-written summary
Tannin pathway key gene expression was over twofold higher with JA than control and MeJA plus SA.
Verbatim
e0“expressions of pathway key genes was more than 2-fold higher in the MeJA and JA treatments than the control and MeJA+SA cells”
t0“grape”
Claim · model-written summary
Stilbenes accumulated in all three treatments.
Verbatim
e0“Stilbenes accumulated in all three treatments”
t0“grape”
Claim · model-written summary
Stilbene levels correlated mostly with STS2 expression.
Not reportedDirection
Verbatim
e0“the levels correlated mostly with the expression of stilbene synthase (STS2)”
e1“Stilbenes accumulated in all three treatments”
t0“grape”
Claim · model-written summary
Several transcription factors had expression patterns matching induction patterns for the three pathways.
Not reportedDirection
Verbatim
e0“several transcription factors that have expression patterns matching the induction patterns seen for the three pathways described above”
t0“grape”
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See all 8 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.