SE50:/MS01

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Sample Set Information

ID SE50
Title Enhancement of oxidative and drought tolerance in arabidopsis by overaccumulation of antioxidant flavonoids
Description We report that flavonoids with radical scavenging activity mitigate against oxidative and drought stress in Arabidopsis thaliana. Metabolome and transcriptome profiling and experiments with oxidative and drought stress in wild-type, single overexpressors of MYB12/PFG1 (PRODUCTION OF FLAVONOL GLYCOSIDES1) or MYB75/PAP1 (PRODUCTION OF ANTHOCYANIN PIGMENT1), double overexpressors of MYB12 and PAP1, transparent testa4 (tt4) as a flavonoid-deficient mutant, and flavonoid-deficient MYB12 or PAP1 overexpressing lines (obtained by crossing tt4 and the individual MYB overexpressor) demonstrated that flavonoid overaccumulation was key to enhanced tolerance to such stresses. Antioxidative activity assays using 2,2 diphenyl-1-picrylhydrazyl, methyl viologen, and 3,3′-diaminobenzidine clearly showed that anthocyanin overaccumulation with strong in vitro antioxidative activity mitigated the accumulation of reactive oxygen species in vivo under oxidative and drought stress. These data confirm the usefulness of flavonoids for enhancing both biotic and abiotic stress tolerance in crops.
Authors Ryo Nakabayashi, Keiko Yonekura-Sakakibara, Kaoru Urano, Makoto Suzuki, Yutaka Yamada, Tomoko Nishizawa, Fumio Matsuda, Mikiko Kojima, Hitoshi Sakakibara, Kazuo Shinozaki, Anthony J. Michael, Takayuki Tohge, Mami Yamazak, Kazuki Saito
Reference Nakabayashi R et al. (2014) The Plant Journal 77: 367-379
Comment


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The raw data files are available at DROP Met web site in PRIMe database of RIKEN.

Analytical Method Details Information

ID MS01
Title LC‒PDA‒MS and LC‒QTOF‒MS
Instrument Waters Acquity UPLC system and Waters Xevo G2 Q-Tof
Instrument Type UPLC-QTOF-MS
Ionization ESI
Ion Mode Positive(LC‒QTOF‒MS)
Description <Flavonoid-targeted profiling by LC‒PDA‒MS>

This analysis was performed as described previously (Tohge et al., 2005a), except that the column was changed to Atlantis T3 (5µm, 4.6 mm×150 mm; Waters, http://www.waters.com/).

Reference:

Tohge, T., Nishiyama, Y., Hirai, M.Y. et al. (2005a) Functional genomics by integrated analysis of metabolome and transcriptome of Arabidopsis plants over-expressing an MYB transcription factor. Plant J. 42, 218–235.


<Untargeted profiling by LC‒QTOF‒MS>

Frozen or freeze-dried samples were extracted with 5 or 50 μl of 80% methanol (MeOH) containing 2.5 μm lidocaine per milligram fresh or dry weight, respectively, which was an internal standard for positive mode analysis, using a mixer mill (MM300; Retsch, http://www.retsch.com/) together with one zirconia bead per tube for 10 min at 20 Hz. After centrifugation at 15 000 g for 10 min and filtration using an Oasis HLB 96-well μElution Plate (Waters), the extracts (1 μl) were analyzed by LC‒QTOF‒MS (LC, Waters Acquity UPLC system; MS, Waters Xevo G2 Q-Tof). The analytical conditions were as follows. For LC: column, Acquity bridged ethyl hybrid (BEH) C18 (1.7 μm, 2.1 mm × 100 mm, Waters); solvent system, solvent A (water including 0.1% formic acid) and solvent B (acetonitrile including 0.1% formic acid); gradient program, 99.5% A/0.5% B at 0 min, 99.5% A/0.5% B at 0.1 min, 20% A/80% B at 10 min, 0.5% A/99.5% B at 10.1 min, 0.5% A/99.5% B at 12.0 min, 99.5% A/0.5% B at 12.1 min, and 99.5% A/0.5% B at 15.0 min; flow rate, 0.3 ml min−1; column temperature, 40°C. For MS: capillary voltage, +3.0 keV, cone voltage, 25.0 V; source temperature, 120°C; desolvation temperature, 450°C; cone gas flow, 50 L h−1; desolvation gas flow, 800 L h−1; collision energy, 6 V; mass range, m/z 100–1500; scan duration, 0.1 sec; interscan delay, 0.014 sec; mode, centroid; polarity, positive; lockspray (leucine enkephalin); scan duration, 1.0 sec; interscan delay, 0.1 sec.

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