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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

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Sample Information

Title Arabidopsis thaliana
Organism - Scientific Name Arabidopsis thaliana
Organism - ID NCBI taxonomy:3702
Compound - ID
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.
Sample Preparation Details ID

Analytical Method Information

Title LC-MS
Method Details ID MS1
Sample Amount

Analytical Method Details Information

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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Data Analysis Information

Title Metabolites identification
Data Analysis Details ID DS1
Recommended decimal places of m/z

Data Analysis Details Information

Title Metabolites identification
Description Metabolites were identified based on UV/visible absorption spectra, quasi-molecular ions and MS/MS information by tandem MS analysis in comparison with the reported anthocyanins (Bloor & Abrahams 2002, Tohge et al. 2005).

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