SE195:/S10/M3
Sample Set Information
ID | SE195 |
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Title | LC-MS analysis of LIPID REMODELLING REGULATOR 1 (LRL1) transgenic lines of Chlamydomonas reinhardtii |
Description | A metabolome analysis was conducted by LC-MS using a wild type Chlamydomonas reinhardtii (C9) and a paromomycin insertional LIPID REMODELLING REGULATOR 1 (LRL1) mutant line (lrl1-1) grown in phosphorus (P)-replete and P-deplete conditions for 2, 4 and 8 days. |
Authors | Nur Akmalia Hidayati, Yui Yamada-Oshima, Masako Iwai, Takashi Yamano, Masataka Kajikawa, Nozomu Sakurai, Kunihiro Suda, Kanami Sesoko, Koichi Hori, Takeshi Obayashi, Mie Shimojima, Hideya Fukuzawa, Hiroyuki Ohta |
Reference | LIPID REMODELLING REGULATOR 1 (LRL1) is differently involved in the phosphorus-depletion response from PSR1 in Chlamydomonas reinhardtii. The Plant Journal (in press) |
Comment |
Sample Information
ID | S10 |
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Title | TAP medium |
Organism - Scientific Name | |
Organism - ID | |
Compound - ID | |
Compound - Source | |
Preparation | Freshly prepared TAP medium was used. |
Sample Preparation Details ID | SS1 |
Comment |
Sample Preparation Details Information
ID | SS1 |
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Title | C. reinhardtii strains and culture conditions |
Description | The C. reinhardtii strain C9 (CC-408, wild type mt-) and lrl1-1 were obtained from the Kyoto University C. reinhardtii mutant library. Screening of lrl1-1 from the Chlamydomonas mutant library was performed according to Gonzalez-Ballestar et al. 2014 . Briefly, several primers spanning the upstream region to the 3' end region of Cre03.g197100 were designed. The superpool library was used as the PCR template, and any amplified product from a primer pair that included a specific region of Cre03.g197100 and AphVIII were sequenced, and its positional tag was identified. Further DNA sequencing indicated that AphVIII-tag insertion was in the upstream region of Cre03.g197100, 115 bp upstream of the 5' untranslated region (UTR), which is predicted to be the promoter region. Liquid cultures were grown in an Erlenmeyer flask mixotrophically in Tris-acetate-phosphate (TAP) medium. For all cultures, cells were cultivated under continuous illumination at 20-40 umol m^-2 s^-1 and 100 rpm shaking at 25C. To induce P-starvation, mid-log phase cells (3-5 x10^6 cells/ml) were centrifuged at 2000 rpm for 5 min and washed twice in a P-free (TAP-P) medium. Potassium phosphate was replaced by 1.5 mM KCl in TAP-P medium. Cells were initially adjusted to 1x10^5 cells/ml on culture day 0. Cultured cells were collected by centrifugation, immediately frozen in liquid nitrogen, and stored at -80C until use. |
Comment_of_details |
Analytical Method Information
ID | M3 |
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Title | Method 1: ESI Positive, Data dependent MS2 scan (FT, IT) |
Method Details ID | MS1 |
Sample Amount | 5 uL medium/ 20 uL injection |
Comment |
Analytical Method Details Information
ID | MS1 |
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Title | LC-FT-MS, ESI, Positive (method 1) |
Instrument | Agilent1100 HPLC (Agilent), LTQ-FT (Thermo Fisher Scientific) |
Instrument Type | LC-FTICR-MS |
Ionization | ESI |
Ion Mode | Positive |
Description | - Sample Extraction
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Comment_of_details | [column] TSK-gel ODS-100V (4.6 x 250 mm, 5 micrometer; TOSOH)
[gradient] Solvent A: water containing 0.1% v/v formic acid; Solvent B: acetonitrile containing 0.1% v/v formic acid; Gradient: 3% B (0 min), 97% B (90 min), 97% B (100 min), 3% B (100.1 min), and 3% B (107 min) [total separation time] 107 min |