GC–MS-Based Nontargeted and Targeted Metabolic Profiling Identifies Changes in the Lentinula edodes Mycelial Metabolome under High-Temperature Stress
Abstract
:1. Introduction
2. Results
2.1. Physiological Study of Different Strains of L. edodes in Response to High-Temperature Stress
2.2. GC/MS Detection Results and Confirmation by Nontargeted Metabonomics
2.3. Identification of Compounds
2.4. Data Dimension Reduction Processing Result
2.5. Screening of Differential Metabolites and Pathway Enrichment Analysis
2.6. Pathway Analysis of Significantly Differential Metabolites
2.7. Targeted Metabolomic Analysis of Key Metabolites
3. Discussion
3.1. Physiological Study on Different Strains of L. edodes in Response to High-Temperature Stress
3.2. Metabonomic Study on Different Strains of L. edodes in Response to High-Temperature Stress
4. Materials and Methods
4.1. Materials and Design
4.1.1. Test Materials
4.1.2. Mycelial Culture and High-Temperature Stress Treatment
4.2. Physiological Determination of Mycelia under High-Temperature Stress
4.2.1. Morphology and Growth Rate of Mycelia
4.2.2. A Method for Determining the Conductivity of Mycelia Damaged by High Temperature
- (1)
- The cultured mycelia were filtered with nonwoven fabric and then rinsed with 200 mL double-distilled water, and the mycelial balls were placed into a triangular bottle again.
- (2)
- The cultured mycelial balls were treated by heat shock for 0 h, 4 h, 8 h, 12 h, 18 h, or 24 h. Then, the treated mycelial balls were put into 30 mL double-distilled water. At room temperature, the conductivity of the solution was determined to be E1.
- (3)
- The centrifuge tube used in the conductivity measurements was sterilized at 121 °C for 20 min and shaken on a shaker at room temperature at 150 rpm for 24 h. The conductivity was measured at E2 at room temperature.
- (4)
- Based on the following formula, the relative conductivity was determined as E% = (E1−E0)/(E2−E0), where E0 refers to the conductivity of double-distilled water [27].
4.2.3. Determination of the Content of Malondialdehyde (MDA) in L. edodes Mycelia under High-Temperature Stress
4.3. Chemicals
4.4. Sample Preparation
4.5. GC/MS Analysis
4.6. Data Preprocessing and Statistical Analysis
4.7. Selection of Differential Metabolites
4.8. Determination of Targeted Metabolites in Key Heat Stress Substances
4.8.1. Preparation of Standard Solution
4.8.2. Sample Treatment Shown in Section 4.4
4.8.3. GC–MS Analysis Method Shown in Section 4.5
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Different Metabolites | VIP | p | Different Metabolites | VIP | p |
---|---|---|---|---|---|
Alanine | 1.17812 | 4.88607 × 10−4 | Gallic acid | 1.30530 | 6.52562 × 10−5 |
Aspartate/aspartic acid | 1.03574 | 3.17468 × 10−7 | Ethanolamine | 1.38919 | 4.47419 × 10−10 |
Glutamate/L-glutamic acid | 1.35766 | 3.41897 × 10−7 | 2-aminoethanethiol | 1.35147 | 1.89102 × 10−6 |
Proline | 1.03591 | 2.860688 × 10−3 | Dithioerythritol | 1.38845 | 2.77433 × 10−7 |
Phenylalanine | 1.07183 | 7.741247 × 10−3 | Phytanic acid | 1.40061 | 1.16715 × 10−7 |
Tyrosine | 1.33479 | 4.72696 × 10−4 | Adenine | 1.38915 | 8.3489 × 10−7 |
Serine | 1.46222 | 1.20382 × 10−5 | Adenosine | 1.16933 | 7.23195 × 10−4 |
L-glutamic acid | 1.02125 | 7.3379 × 10−5 | D-Fructose 1,6-bisphosphate | 1.17656 | 5.699884 × 10−3 |
Glycine | 1.22217 | 2.05583 × 10−8 | Fructose-6-phosphate | 1.36802 | 1.73526 × 10−4 |
Threonine | 1.17647 | 2.305898 × 10−3 | Alpha-ketoglutaric acid | 1.38852 | 2.1916 × 10−6 |
Glutathione | 1.25819 | 1.36582 × 10−12 | 1,3-diaminopropane | 1.40000 | 4.07715 × 10−8 |
Citrate cycle | 1.38953 | 6.38001 × 10−10 | 1-Methylhydantoin | 1.34342 | 2.69444 × 10−6 |
Fructose | 1.22170 | 5.66071 × 10−8 | 3,6-Anhydro-D-galactose | 1.44528 | 7.65583 × 10−12 |
Maltose | 1.24022 | 3.8289 × 10−13 | 5-Methoxytryptamine | 1.42654 | 3.86987 × 10−9 |
Sophorose | 1.23167 | 4.39968 × 10−7 | Acetol | 1.37941 | 4.87405 × 10−8 |
Trehalose | 1.07189 | 2.184293 × 10−3 | Allylmalonic acid | 1.30494 | 2.91429 × 10−5 |
Sorbitol | 1.32470 | 1.68173 × 10−5 | Cycloleucine | 1.36591 | 2.25922 × 10−4 |
3-Hydroxypropionic acid | 1.41517 | 5.02333 × 10−9 | Dibenzofuran | 1.31198 | 5.32826 × 10−6 |
5-Aminovaleric acid | 1.42903 | 2.50953 × 10−9 | Gly-pro | 1.40607 | 1.92801 × 10−8 |
Aminomalonic acid | 1.33618 | 3.96316 × 10−6 | N-Acetyl-D-galactosamine | 1.03300 | 4.551565 × 10−3 |
Dehydroascorbic Acid | 1.42405 | 5.98067 × 10−6 | Octanal | 1.39123 | 4.41212 × 10−7 |
Oxalic acid | 1.27637 | 1.62664 × 10−9 | Panthenol | 1.25303 | 7.58015 × 10−4 |
Trehalose-6-phosphate | 1.24712 | 4.56643 × 10−6 | p-benzoquinone | 1.35201 | 2.09886 × 10−6 |
Mannitol | 1.22484 | 1.58142 × 10−11 |
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Zhao, X.; Chen, M.; Zhao, Y.; Zha, L.; Yang, H.; Wu, Y. GC–MS-Based Nontargeted and Targeted Metabolic Profiling Identifies Changes in the Lentinula edodes Mycelial Metabolome under High-Temperature Stress. Int. J. Mol. Sci. 2019, 20, 2330. https://doi.org/10.3390/ijms20092330
Zhao X, Chen M, Zhao Y, Zha L, Yang H, Wu Y. GC–MS-Based Nontargeted and Targeted Metabolic Profiling Identifies Changes in the Lentinula edodes Mycelial Metabolome under High-Temperature Stress. International Journal of Molecular Sciences. 2019; 20(9):2330. https://doi.org/10.3390/ijms20092330
Chicago/Turabian StyleZhao, Xu, Mingjie Chen, Yan Zhao, Lei Zha, Huanling Yang, and Yuejin Wu. 2019. "GC–MS-Based Nontargeted and Targeted Metabolic Profiling Identifies Changes in the Lentinula edodes Mycelial Metabolome under High-Temperature Stress" International Journal of Molecular Sciences 20, no. 9: 2330. https://doi.org/10.3390/ijms20092330