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MB Sample ID: SA094164
Local Sample ID: | 10_18_2011 M3C |
Subject ID: | SU001370 |
Subject Type: | Plant |
Subject Species: | Corylus avellana;Corylus americana |
Taxonomy ID: | 13451;78632 |
Genotype Strain: | Corylus avellana;Corylus americana(cv Geneva) |
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Subject:
Subject ID: | SU001370 |
Subject Type: | Plant |
Subject Species: | Corylus avellana;Corylus americana |
Taxonomy ID: | 13451;78632 |
Genotype Strain: | Corylus avellana;Corylus americana(cv Geneva) |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
10_18_2011 M3C | SA094164 | FL013459 | 460 | FeEDDHA (µM) |
Collection:
Collection ID: | CO001365 |
Collection Summary: | Cultures were sampled in triplicate on day 35 of the culture period, |
Sample Type: | Plant |
Treatment:
Treatment ID: | TR001385 |
Treatment Summary: | Filbert (C. avellana L. × C. americana Marshall cv. Geneva; Grimo Nut Nursery, Niagara-on-the-Lake, ON, Canada) shoot cultures were provided from germplasm maintained at the Gosling Research Institute for Plant Preservation (GRIPP; University of Guelph, Guelph, ON). Plantlets were grown in GA-7 vessels according to methods previously described (Garrison et al. 2013; Latawa et al. 2016). In brief, cultures were grown on semi-solid modified NCGR-COR medium (Yu and Reed 1995) supplemented with 10 g L-1 myo-inositol, 200 mg L-1 glycine, 100 mg L-1 nicotinic acid, 100 mg L-1 thiamine (PhytoTechnology Laboratories), 17.6 µM benzylaminopurine (BAP; Sigma-Aldrich), 0.014 µM indole-3-butyric acid (IBA; Sigma-Aldrich), 0.29 µM gibberellic acid (GA3; PhytoTechnology Laboratories), and 30 g L-1 glucose with 0, 230, 460 and 690 µM Fe-EDDHA (Sigma, St. Louis, MO) and the pH of the medium was adjusted to 5.7 before autoclaving at 121 ºC for 20 min. Cultures were maintained in a growth room at 23 ± 2oC with a 16 h photoperiod of 40 µmol m-2 s-1 provided by cool-white fluorescent lamps (Osram Sylvania Ltd., Mississauga, ON, Canada). |
Sample Preparation:
Sampleprep ID: | SP001378 |
Sampleprep Summary: | Cultures were sampled in triplicate on day 35 of the culture period, accurately weighed (50 mg), and homogenized in 1 mL of 70% ethanol for 30 s (Kontes Pellet Pestle, Fisher Scientific). Samples were centrifuged (16,000 x g) for 3 min to settle particulate matter and the supernatant was filtered (0.1 µm, Ultrafree-MC filtered centrifuge tubes; Millipore, MS, USA) prior to chromatography. |
Combined analysis:
Analysis ID | AN002157 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity |
Column | Waters BEH Acquity C18 (150 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Waters Micromass QTOF Premier |
Ion Mode | POSITIVE |
Units | Peak Intensity |
Chromatography:
Chromatography ID: | CH001575 |
Chromatography Summary: | Extracts and 70% ethanol blanks (n=3 for each treatment) were separated using a Waters BEH Acquity C18 (2.1 X 150 mm, 1.7 µm) column with the following gradient: 0.1% aqueous formic acid:acetonitrile (0.0-25 min, 95:5-5:95 v/v, 25.01-30.0 min, 95:5 v/v). The flow rate was set to 0.25 mL min-1 for 30 mins at 30 ◦C (Waters Acquity UPLC). |
Instrument Name: | Waters Acquity |
Column Name: | Waters BEH Acquity C18 (150 x 2.1mm,1.7um) |
Column Temperature: | 30 |
Flow Gradient: | 0.0 - 10.0 min, : 95:5-5:95 v/v, 10.0-15.0 min, 5:95 v/v, 15.0-20.0min, 5:95-95:5 v/v, 20.0-25.0min, 95:5 v/v) |
Flow Rate: | 0.25 ml/min |
Solvent A: | 100% water; 1% formic acid |
Solvent B: | 100% acetonitrile |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002006 |
Analysis ID: | AN002157 |
Instrument Name: | Waters Micromass QTOF Premier |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | A steady flow of leucine enkephalin (Waters 1525 HPLC binary solvent manager, 2 ng mL-1) was used as the internal standard for calibration of the Micromass LCT Premier series ToF-MS (Waters Inc.). Time of flight mass spectrometry was used with previously published optimized conditions (Brown, Murch, et al. 2012; Brown et al. 2012) including: electrospray ionization and positive ion detection in W mode, mass range of 100-1000 amu and a scan time of 0.1s. Data were collected with MassLynx V4.1 and exported via MarkerLynx. Data were processed in Excel™ to align retention times and remove multiply charged ions as described previously (Brown et al. 2012; Brown et al. 2012; Turi and Murch 2013; Turi et al. 2014). |
Ion Mode: | POSITIVE |