SE193:/S1/M2

Sample Set Information

ID	TSE1352
Title	Metabolic Reprogramming in Leaf Lettuce Grown Under Different Light Quality and Intensity Conditions Using Narrow-Band LEDs
Description	Light-emitting diodes (LEDs) are an artificial light source used in closed-type plant factories and provide a promising solution for a year-round supply of green leafy vegetables, such as lettuce (Lactuca sativa L.). Obtaining high-quality seedlings using controlled irradiation from LEDs is critical, as the seedling health affects the growth and yield of leaf lettuce after transplantation. Because key molecular pathways underlying plant responses to a specific light quality and intensity remain poorly characterised, we used a multi-omics–based approach to evaluate the metabolic and transcriptional reprogramming of leaf lettuce seedlings grown under narrow-band LED lighting. Four types of monochromatic LEDs (one blue, two green and one red) and white fluorescent light (control) were used at low and high intensities (100 and 300 μmol·m−2·s−1, respectively). Multi-platform mass spectrometry-based metabolomics and RNA-Seq were used to determine changes in the metabolome and transcriptome of lettuce plants in response to different light qualities and intensities. Metabolic pathway analysis revealed distinct regulatory mechanisms involved in flavonoid and phenylpropanoid biosynthetic pathways under blue and green wavelengths. Taken together, these data suggest that the energy transmitted by green light is effective in creating a balance between biomass production and the production of secondary metabolites involved in plant defence.
Authors	Kazuyoshi Kitazaki, Atsushi Fukushima, Ryo Nakabayashi, Yozo Okazaki, Makoto Kobayashi, Tetsuya Mori, Tomoko Nishizawa, Sebastian Reyes-Chin-Wo, Richard W. Michelmore, Kazuki Saito, Kazuhiro Shoji & Miyako Kusano
Reference	Scientific Reports, volume 8, Article number: 7914 (2018)
Comment

www.nature.com/articles/s41598-018-25686-0

Sample Information

ID	S1
Title	red leaf lettuce
Organism - Scientific Name	Lactuca sativa L.
Organism - ID	NCBI:txid4236
Compound - ID
Compound - Source
Preparation	BioSource Species Lactuca sativa L. Genotypes/Varieties cv. Banchu Red Fire Organ specification The third leaf Growth conditions Under white fluorescent light (FL, FLR110H-W1A; Mitsubishi/Osram Co.; Yokohama, Japan), seeds of red-leaf lettuce (Lactuca sativa L. cv Banchu red fire; Takii seed, Kyoto, Japan) were pregerminated [14 hr, 14 days, 23 ± 2ºC, 100 µmol m−2 s−1 photon synthetic photon flux density (PPFD)]. The seedlings were supplied with a nutrient solution (Otsuka hydroponic composition, Otsuka Chemical Co. Ltd., Osaka, Japan) adjusted to an electrical conductivity (EC) of 1.2 dS/m and pH 5.8. It contained 7.0 mmol l-1 NO3-, 0.6 mmol l-1 NH4+, 3.7 mmol l-1 K+, 2.3 mmol l-1 Ca2+, 1.3 mmol l-1 H2PO4-, and 0.9 mmol l-1 Mg2+. The seedlings were transplanted to cultivation panels in a growth chamber (VB1514; Vötsch, Germany), supplied with nutrient solution for the duration of the experiments, and grown at 25ºC [relative humidity (RH) 60%, 900 µmol mol-1 CO2]. The plants were irradiated with different light spectra from LEDs, namely B470 (peak wavelength 470 nm, ISL305X302-BBBB, CCS Co., Kyoto, Japan), G510 (peak wavelength 510 nm, ISL-305X302- GGGG505, CCS Co.), G520 (peak wavelength of 524 nm; ISL-305X302-GGGG525, CCS) and R680 (peak wavelength of 680nm; ISL-305X302-RRRR68, CCS Co., Kyoto, Japan). The seedlings were irradiated for 24 hr at PPFD 100 or 300 µmol m−2 s−1 (P100, P300). The wavelength of the light source was determined with a USB2000 spectrometer (Ocean Optics, Dunedin, FL, USA). At 14, 15 and 21 DAS, dry weight (DW) was measured.
Sample Preparation Details ID
Comment

Analytical Method Information

ID	M2
Title	LC-q-TOF-MS (to detect secondary metabolites)
Method Details ID	MS2
Sample Amount	1 μL
Comment

Analytical Method Details Information

ID	MS2
Title	LC-q-TOF-MS (to detect secondary metabolites)
Instrument	LC, Waters Acquity UPLC system; MS, Waters Xevo G2 Q-Tof
Instrument Type
Ionization	ESI
Ion Mode	negative
Description	Extraction for LC-q-TOF-MS to detect secondary metabolites Each frozen sample was extracted with 5 fold amount of solvent (methanol/water [8:2 v/v]) containing a reference compound (2.5 µM of 10-camphorsulfonic acid ([M-H]-, m/z 231.0691)) using a mixer mill MM301 (Retsch) at a frequency of 18 Hz for 7 min at 4°C. After centrifugation for 10 min at 17,000 × g, the supernatant was filtered using an Oasis® HLB µElusion plate (30 µm, Waters Co., Massachusetts, US). LC-q-TOF-MS conditions to detect secondary metabolites After preparation of the extracts, the sample extracts (1 µl) were analyzed using an LC-MS system equipped with an electrospray ionization (ESI) interface (LC, Waters Acquity UPLC system; MS, Waters Xevo G2 Q-Tof). The analytical conditions were as follows. LC: column, Acquity bridged ethyl hybrid (BEH) C18 (pore size 1.7 µm, length 2.1× 100 mm, Waters); solvent system,acetonitrile(0.1% formic acid):water (0.1% formic acid); gradient program, 0.5 : 99.5 v/v at 0 min, 0.5 : 99.5 v/v at 0.1 min, 80 : 20 v/v at 10 min, 99.5 : 0.5 v/v at 10.1 min, 99.5 : 0.5 v/v at 12 min and 0.5 : 99.5 v/v at 12.1 min, 0.5 : 99.5 v/v at 15 min; flow rate, 0.3 ml/min; temperature, 40 °C; MS detection: capillary voltage, +3.0 keV, cone voltage, 25.0 V, source temperature, 120°C, desolvation temperature, 450 °C, cone gas flow, 50 l/ h; desolvation gas flow, 800 l/h; collision energy, 6 V; mass range, m/z 50‒1500; scan duration, 0.1 sec; interscan delay, 0.014 sec; mode, centroid; polarity, negative; Lockspray (Leucineenkephalin): scan duration, 1.0 sec; interscan delay, 0.1 sec. The data were recorded using Progenesis CoMet (Nonlinear Dynamics).
Comment_of_details