| S Preparation
|
BioSource Species: <br />Arabidopsis … BioSource Species: <br />Arabidopsis thaliana L. cv. Columbia ecotype 0 (wild-type)
Genotype: Wild-type, transparent-testa-4 [tt4] (Shikazono et al., 2001), transparent-testa-5 [tt5] (SALK_034145) and sinapoylglucose-accumulator-1 [sng1] (Lorenzen et al., 1996) in Col-0 background<br /><br />
Organ:<br />Aerial regions<br /><br />
Organ specification:<br />18-day-old aerial parts were harvested.<br /><br />
Growth condition:<br />The sterilized seeds were stratified at 5°C for 2 d, and were successively sown on Murashige and Skoog (MS) medium (Wako, Osaka, Japan) containing 1% sucrose. These four genotypes were grown, side-by-side, under highly controlled growth conditions, under five independent environmental regimes. The chosen conditions were; (i) growth for 18 days under typical long day conditions (16h light/ 8h dark photoperiod; LC), (ii) growth for 17 days under typical long day conditions with 8 h additional light on day 18 (continuous light 1; CO1), (iii) growth for 17 days under typical long day conditions with 24-h ultraviolet light on day 18 (ultraviolet light 1; UV1), (iv) growth for 14 days under typical long day conditions followed by four days (96h) of continuous light (continuous light 4; CO4) and (v) growth for 14 days under typical long day conditions followed by four days of ultraviolet light (ultraviolet light 4; UV4). Plants were cultivated in controlled growth chambers (SANYO; MLR-350) at 22°C in 16-h light and 8-h dark conditions for 14 days (for CO4 and UV4), 17 days (for CO1 and UV1) and 18 days (for LC). The aerial regions were harvested, 6 h after the onset of the bright phase under each light treatment. Among these harvested plants, the plants that had fresh weight over 10 mg were used for metabolite profiling (9-11 different biological replicates for each condition).<br /><br />
Experimental condition:<br />See the growth conditions.<br /><br />
Light condition: 50 μmol m-2 s-1 for LC, 60 μmol m-2 s-1 for CO1 and CO4; 830 mW m-2 s-1 for UV1 and UV 4 irradiated by narrow-band (312 nm) ultraviolet-B light (CSL-30B, COSMO BIO).<br /><br />
Growth plot design:<br />Plates were randomized every third day in each growth chamber.
Sampling and sampling date: The aerial regions were harvested in February 2007 (5th, 6th and 7th February, 2007) for metabolite profiling, in July and August 2009 for qRT-PCR and microarray analysis.<br /><br />
Metabolism quenching method:<br />All the plant materials were frozen immediately (within 30 sec after sampling) in liquid nitrogen to quench the enzymatic activity. Samples were stored at -80°C until use. y. Samples were stored at -80°C until use.
|
