Flasks with N. crassa wild type strain (FGSC #2489) were pre-grown for 16 h on 2% sucrose medium, then washed three times with 1× Vogel’s salt solution (no carbon source added) for a total duration of 30 min before transferring to their respective carbon source: 1 mM galacturonic acid (D -GalA; Sigma Aldrich) plus 1 mM rhamnose (L- Rha; Sigma Aldrich), referred to as GalAR (D -GalA + L -Rha), 2 mM glucose (D -Glc; Sigma Aldrich), 2 mM xylose (D -Xyl; Sigma Aldrich), 0.5% cellobiose (Cel; Sigma Aldrich) and 2 mM 1,4-ß-D -glucosyl-D -mannose plus 1,4-ß-D -mannobiose (referred to as Glucomannodextrins or Gm; Megazyme). Samples for global proteome and phosphoproteome were incubated for additional 2 min. No carbon condition (NC) was incubated for 1 h after the medium switch (Figure 1A ). More details about the experimental methodology for Mass Spectrometry can be found in the Supplementary Material (Supplementary Table S1) .
D gala
Manufactured by Merck Group
Sourced in United States
D-GalA is a laboratory reagent that is produced by Merck Group. It is a monosaccharide that is commonly used in various chemical and biochemical applications. The core function of D-GalA is to serve as a building block for the synthesis of larger carbohydrate molecules and to facilitate various analytical and research procedures.
Automatically generated - may contain errors
Lab products found in correlation
D glc, by Merck Group (4 mentions)
L rha, by Merck Group (3 mentions)
D gal, by Merck Group (3 mentions)
D xyl, by Merck Group (3 mentions)
D glca, by Merck Group (2 mentions)
L ara, by Merck Group (2 mentions)
Carbopac pa20 column, by Thermo Fisher Scientific (2 mentions)
L fuc, by Merck Group (2 mentions)
Dionex ics 5000, by Thermo Fisher Scientific (1 mentions)
Pulsed amperometric detector, by Thermo Fisher Scientific (1 mentions)
Dionex ics 2500, by Thermo Fisher Scientific (1 mentions)
D fru, by Merck Group (1 mentions)
Faststart universal sybr green master rox kit, by Roche (1 mentions)
Improm 2 reverse transcription system, by Promega (1 mentions)
2 2 diphenyl 1 picrylhydrazyl dpph free radical, by Merck Group (1 mentions)
Mouse il 6 elisa kit, by Thermo Fisher Scientific (1 mentions)
5 protocols using d gala
1
Proteomic Analysis of N. crassa Responses to Carbon Sources
2
Quantifying Non-Cellulosic Polysaccharides
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The composition of non-cellulosic polysaccharides was determined according to Foster et al. (2010) with modifications. Due to shortage of plant material, analysis was performed only for samples harvested at the heading stage and maturity. De-starched AIR material (5 mg) was hydrolysed in 2 M TFA for 90 min at 121 °C. TFA was removed by drying under vaccum. TFA breaks down the cell wall network to release primarily non-cellulosic polysaccharides, and some fractions of cellulose that contain kinks and chain dislocations (amorphous form). The composition of monosaccharides in the filtrate was determined and quantified by high-performance anion-exchange chromatography coupled with integrated pulsed amperometric detection (HPAEC-iPAD) using Thermo Scientific Dionex ICS5000 as described previously by Głazowska et al. (2018) (link). The system was calibrated with standards (l -Ara, d -Xyl, d -Gal, d -Glc, d -GlcA, and d -GalA) (Sigma). All calculations were done in Chromeleon CDS software.
3
Monosaccharide Composition Analysis of Plant Cell Walls
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To determine monosaccharide composition, 3 replicates of cell walls from 10 plants were used. 1 mg of dry de-starched cell wall was hydrolyzed with 2 N trifluoroacetic acid at 120 °C for 2 h. The hydrolysates were dried at 50 °C, re-dissolved in water, and analyzed by high-performance anion-exchange chromatography with pulsed-amperometric detection using a CarboPac PA-20 column (3 mm × 150 mm; Dionex, Sunnyvale, CA, USA) as described earlier [99 (link)]. Monosaccharides were separated using a gradient of 100 mM NaOH in water at 0.5 mL min−1 under the following conditions: 0–0.05 min—12 mM NaOH; 0.05–26 min—0.65 mM NaOH; 26–46 min—300 mM NaOH; 46–55 min—12 mM NaOH. Monosaccharide standards included L-Fuc, L-Rha, L-Ara, D-Gal, D-Glc, D-Xyl, D Man, D-GalA, and D-GlcA (all from Sigma–Aldrich, St. Louis, MO, USA). To determine response factors, standard curves were created using mixtures of all standard monosaccharides at different concentrations.
4
Monosaccharide Composition Analysis of Polysaccharides
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Samples (2 mg) of six polysaccharide components were hydrolyzed with 3 mL 2 M trifluoroacetic acid (TFA) at 110 °C for 4 h. After hydrolysis, hydrolysates were dried with a Termovap sample concentrator, and then 3 mL methanol was added and dried repeatedly three times until the TFA was completely removed. The monosaccharide compositions were determined by a high-performance anion exchange chromatography (HPAEC) system (Dionex ICS-2500, Dionex, Sunnyvale, CA, USA) equipped with a CarboPac™ PA20 column (3 mm × 150 mm, Dionex, USA) and a pulsed amperometric detector (Dionex, USA). The column was eluted with 2 mM NaOH (0.45 mL/min) followed by 0.05 to 0.2 M NaAc at 30 °C. The monosaccharide compositions and content of polysaccharide components were determined using d -Gal, d -Glc, d -Ara, l -Fuc, l -Rha, d -Man, d -Xyl, d -Fru, d -Rib, d -GluA, and d -GalA (Sigma-Aldrich, St. Louis, MO, USA) as the standards.
5
Extraction and Characterization of S. ningpoensis Roots
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The roots of S. ningpoensis were collected from the experimental field of Jining Medical University (Rizhao, Shandong, China) and identified by Jian'an Wang (Medical Botanist, Jining Medical University). Monosaccharide standards D-glucose (D-Glc), D-GalActose (D-Gal), D-arabinose (D-Ara), L-fucose (L-Fuc), D-mannose (D-Man), L-rhamnose (L-Rha), D-fructose (D-Fru), D-xylose (D-Xyl), D-GlcA, and D-GalA were purchased from Sigma-Aldrich (St. Louis, MO, USA). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical was purchased from Sigma-Aldrich (St. Louis, MO, USA). 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical was purchased from the Shanghai Yuanye Biotechnology Co. Ltd. (Shanghai, China). ImProm-II Reverse Transcription System was obtained from Promega Corporation (Madison, WI, USA). IL-6 Mouse ELISA Kit was purchased from Invitrogen (Carlsbad, CA, USA). TNF-α Mouse ELISA Kit was obtained from PeproTech (Rocky Hill, NJ, USA). UNIQ-10 column Trizol total RNA extraction kit was obtained from Sangon Biological Engineering Technology and Services Co., Ltd. (Shanghai, China). FastStart Universal SYBR Green Master (ROX) kit was purchased from Roche (Mannheim, Germany). All chemical reagents were of analytical grade, and the deionized water was used in the whole experiment.
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