Abstract
(Iso)flavonoids are commonly accumulated as malonylated or acetylated glycoconjugates in legumes. Sequence analysis on EST database of the model legume Medicago truncatula enabled us to identify nine cDNA sequences encoding BAHD super-family enzymes that are distinct from the most of the characterized anthocyanin/flavonol acyltransferase genes in other species. Functional characterization revealed that three of these corresponding enzymes, MtMaT1, 2 and 3, specifically recognize malonyl CoA as an acyl donor and catalyze the malonylation of a range of isoflavone 7-O-glucosides in vitro. These malonyltransferase genes displayed distinct tissue-specific expression patterns and responded differentially to biotic and abiotic stresses. Consistent with gene expression, the level of the accumulated malonyl isoflavone glucoside was altered in the roots of M. truncatula grown under normal and drought-stressed conditions. Overexpression of the MtMaT1 gene in a previously engineered Arabidopsis line that accumulates genistein glycosides (Proc. Natl Acad. Sci. USA, 99, 2002:14578) led to a malonylated product. Confocal microscopy of the transiently expressed MtMaT1-GFP fusion revealed strong fluorescence in both the cytoplasm and nucleus of M. truncatula and tobacco leaf cells. A truncated MtMaT1 lacking the C-terminal polypeptide of 110 amino acid residues that include the DFGWG motif, the single conserved sequence signature of BAHD super-family members, retained considerable catalytic efficiency, but showed an altered optimum pH preference for maximum activity. Such C-terminal polypeptide deletion or deletion of the DFGWG motif alone led to improper folding of the transiently expressed GFP fusion protein in living cells, and impaired nuclear localization of the enzyme.
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Claims 14
Claim · model-written summary
Nine distinct BAHD enzyme cDNA sequences were identified.
- Taxon
- Medicago truncatula
Medicago truncatulae0 - Outcome
- cDNA sequences encoding BAHD super-family enzymese0
- Value
- ninee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Medicago truncatula enabled us to identify nine cDNA sequences encoding BAHD super-family enzymes”
Claim · model-written summary
MtMaT1, 2 and 3 use malonyl CoA and malonylate isoflavone 7-O-glucosides in vitro.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Gene
- MtMaT1, 2 and 3e0
- Compound
- malonyl CoAe0
- Outcome
- malonylation of a range of isoflavone 7-O-glucosidese0
- Setting
- in vitroe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“MtMaT1, 2 and 3, specifically recognize malonyl CoA as an acyl donor and catalyze the malonylation of a range of isoflavone 7-O-glucosides in vitro”
t0“Medicago truncatula”
Claim · model-written summary
The malonyltransferase genes had distinct tissue expression patterns.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Gene
- malonyltransferase genese0
- Outcome
- tissue-specific expression patternse0
- Direction
- distincte0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These malonyltransferase genes displayed distinct tissue-specific expression patterns”
t0“Medicago truncatula”
Claim · model-written summary
The genes responded differently to biotic and abiotic stresses.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- biotic and abiotic stressese0
- Gene
- malonyltransferase genese0
- Outcome
- responded differentiallye0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These malonyltransferase genes displayed distinct tissue-specific expression patterns and responded differentially to biotic and abiotic stresses”
t0“Medicago truncatula”
Claim · model-written summary
Root malonyl isoflavone glucoside levels differed under normal and drought stress.
- Taxon
- M. truncatulaMedicago truncatulae0
- Organ / stage
- rootse0
- Treatment
- drought-stressed conditionse0
- Outcome
- level of the accumulated malonyl isoflavone glucosidee0
- Direction
- alterede0
- Comparator
- normale0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the level of the accumulated malonyl isoflavone glucoside was altered in the roots of M. truncatula grown under normal and drought-stressed conditions”
Claim · model-written summary
MtMaT1 overexpression produced a malonylated product in Arabidopsis.
- Taxon
- Arabidopsis
Arabidopsise0 - Treatment
- Overexpression of the MtMaT1 genee0
- Gene
- MtMaT1e0
- Outcome
- malonylated producte0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Overexpression of the MtMaT1 gene in a previously engineered Arabidopsis line that accumulates genistein glycosides (Proc. Natl Acad. Sci. USA, 99, 2002:14578) led to a malonylated product”
Claim · model-written summary
MtMaT1-GFP localized to cytoplasm and nucleus of Medicago leaf cells.
- Taxon
- M. truncatulaMedicago truncatulae0
- Organ / stage
- leaf cellse0
- Gene
- MtMaT1-GFPe0
- Outcome
- strong fluorescence in both the cytoplasm and nucleuse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“MtMaT1-GFP fusion revealed strong fluorescence in both the cytoplasm and nucleus of M. truncatula and tobacco leaf cells”
Claim · model-written summary
MtMaT1-GFP localized to cytoplasm and nucleus of tobacco leaf cells.
- Taxon
- tobacco
Nicotiana tabacume0 - Organ / stage
- leaf cellse0
- Gene
- MtMaT1-GFPe0
- Outcome
- strong fluorescence in both the cytoplasm and nucleuse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“MtMaT1-GFP fusion revealed strong fluorescence in both the cytoplasm and nucleus of M. truncatula and tobacco leaf cells”
Claim · model-written summary
Truncated MtMaT1 retained catalytic efficiency.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- lacking the C-terminal polypeptide of 110 amino acid residuese0
- Gene
- MtMaT1e0
- Outcome
- catalytic efficiencye0
- Direction
- retainede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A truncated MtMaT1 lacking the C-terminal polypeptide of 110 amino acid residues that include the DFGWG motif, the single conserved sequence signature of BAHD super-family members, retained considerable catalytic efficiency”
t0“Medicago truncatula”
Claim · model-written summary
Truncated MtMaT1 had a changed optimal pH.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- lacking the C-terminal polypeptide of 110 amino acid residuese0
- Gene
- MtMaT1e0
- Outcome
- optimum pH preferencee0
- Direction
- alterede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A truncated MtMaT1 lacking the C-terminal polypeptide of 110 amino acid residues that include the DFGWG motif, the single conserved sequence signature of BAHD super-family members, retained considerable catalytic efficiency, but showed an altered optimum pH preference for maximum activity”
t0“Medicago truncatula”
Claim · model-written summary
C-terminal deletion caused improper GFP fusion folding.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- C-terminal polypeptide deletione0
- Outcome
- improper folding of the transiently expressed GFP fusion proteine0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Such C-terminal polypeptide deletion or deletion of the DFGWG motif alone led to improper folding of the transiently expressed GFP fusion protein in living cells”
t0“Medicago truncatula”
Claim · model-written summary
DFGWG motif deletion caused improper GFP fusion folding.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- deletion of the DFGWG motife0
- Outcome
- improper folding of the transiently expressed GFP fusion proteine0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“deletion of the DFGWG motif alone led to improper folding of the transiently expressed GFP fusion protein in living cells”
t0“Medicago truncatula”
Claim · model-written summary
C-terminal deletion impaired nuclear localization.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- C-terminal polypeptide deletione0
- Outcome
- nuclear localization of the enzymee0
- Direction
- impairede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Such C-terminal polypeptide deletion or deletion of the DFGWG motif alone led to improper folding of the transiently expressed GFP fusion protein in living cells, and impaired nuclear localization of the enzyme”
t0“Medicago truncatula”
Claim · model-written summary
DFGWG motif deletion impaired nuclear localization.
- Taxon
- Medicago truncatulaMedicago truncatulat0
- Treatment
- deletion of the DFGWG motife0
- Outcome
- nuclear localization of the enzymee0
- Direction
- impairede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“deletion of the DFGWG motif alone led to improper folding of the transiently expressed GFP fusion protein in living cells, and impaired nuclear localization of the enzyme”
t0“Medicago truncatula”
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