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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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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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