SE191:/S1/M1
From Metabolonote
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Sample Set Information
| ID | TSE1350 |
|---|---|
| Title | Enhanced radical scavenging activity of genetically modified Arabidopsis seeds |
| Description | The proanthocyanidin (PA) content was increased in seeds of pap1-D mutant of Arabidopsis thaliana, in which the expression of endogenous PAP1 gene encoding a Myb-like transcription factor was induced by activation-tagging with enhancer sequences derived from cauliflower mosaic virus 35S promoter. In contrast, the PA contents decreased in seeds of transgenic plants transformed with a PAP1 cDNA or with a PAP1 chimeric repressor, although the amount of soluble anthocyanins increased in seeds of transgenic plants over-expressing PAP1cDNA. The enhanced radical scavenging activity was observed only in the seed extracts of pap1-D mutant, indicating that PAs are primarily responsible for radical scavenging activity in seeds. The present study suggests the feasibility of engineering a transcription factor of flavonoid biosynthesis for health beneficial plant seeds. |
| Authors | Tohge, T., Matsui, K., Ohme-Takagi, M., Yamazaki, M. and Saito, K. |
| Reference | Biotechnol Lett. 2005 Mar;27(5):297-303 |
| Comment |
Sample Information
| ID | S1 |
|---|---|
| Title | Arabidopsis thaliana |
| Organism - Scientific Name | Arabidopsis thaliana |
| Organism - ID | NCBI taxonomy:3702 |
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| Compound - Source | |
| Preparation | The pap1-D mutant of A. thaliana was described previously (Borevitz et al. 2000). The PAP1 cDNA-over-expressing transgenic A. thaliana (PAP1OX) was obtained by transformation with the construct carrying cauliflower mosaicvirus 35S promoter linked with the coding sequence of PAP1 cDNA as described elsewhere (Tohge et al. 2005). The transgenic plant expressing PAP1 chimeric repressor (PAP1SRDX) was described previously (Hiratsu et al. 2003, Matsui et al. 2004). The plants were grown in a standard greenhouse at 22 C in 16 h light for 8 weeks. Seeds were harvested and stored at room temperature for more than 1 week for drying. These seeds were used for analysis of anthocyanins, PAs, and radical-scavenging activity. |
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Analytical Method Information
| ID | M1 |
|---|---|
| Title | LC-MS |
| Method Details ID | MS1 |
| Sample Amount | |
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Analytical Method Details Information
| ID | MS1 |
|---|---|
| Title | LC-MS (for anthocyanin profiling) |
| Instrument | LC: Agilent HPLC 1100 series MS: LCQ-DECA, ThermoQuest |
| Instrument Type | |
| Ionization | ESI |
| Ion Mode | positive |
| Description | Seeds were homogenized in 50 ll extraction solvent (methanol/acetic acid/H2O, 9:1:10 by vol.) per 1 mg fresh tissues in a mixer mill (MM300, Retsch Gmbl & Co. KG, Haan) at 30 rpm. After centrifugation, the supernatant was analyzed by HPLC with photodiode array detector and electrospray ionization mass-spectrometer (ESI-MS) as described previously (Jones et al. 2003; Yamazaki et al. 2003). The eluate from the HPLC (Agilent HPLC 1100 series, Agilent Technologies, Palo Alto, CA) was directly introduced into the mass spectrometer (LCQ-DECA, ThermoQuest, San Jose, CA). |
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