Analysis of the Mechanism of Wood Vinegar and Butyrolactone Promoting Rapeseed Growth and Improving Low-Temperature Stress Resistance Based on Transcriptome and Metabolomics
Abstract
:1. Introduction
2. Results and Analysis
2.1. Wood Vinegar and Butyrolactone Can Promote the Growth, Yield and Quality of Rapeseed
2.2. Wood Vinegar and Butyrolactone Can Improve the Low-Temperature Tolerance of Rapeseed
2.3. Transcriptome Analysis of Rapeseed Treated with Wood Vinegar and Butyrolactone
2.4. qPCR Validation of Transcriptome Data
2.5. Identification and Analysis of Differential Metabolites
2.6. Differential Gene and Differential Metabolite Pathway Analysis
3. Materials and Methods
3.1. Plant Materials and Growth Conditions
3.2. Spraying Treatment and Sampling
3.3. Determination of Morphology and Physiology
3.4. Determination of Yield and Quality
3.5. RNA Isolation and Library Construction
3.6. RNA Sequencing Data Analysis
3.7. qRT PCR Analysis
3.8. Metabolite Extraction
3.9. LC-MS Analysis
3.10. Metabolite Data Analysis
3.11. Association Analysis of Transcriptome and Metabolome
3.12. Data Processing and Analysis
4. Discussion
4.1. Wood Vinegar and Butyrolactone Can Improve the Yield of Rapeseed
4.2. Physiological Changes in Rapeseed in Response to Low-Temperature Stress Enhanced by Wood Vinegar and Butyrolactone
4.3. Transcriptome Analysis of Wood Vinegar and Butyrolactone on Rapeseed
4.4. Metabolomic Analysis of Wood Vinegar and Butyrolactone on Rapeseed
4.5. Integrated Transcriptional and Metabolomic Analysis of KEGG Pathway of Wood Vinegar and Butyrolactone on Rapeseed
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Treatment | Root Collar Diameter (mm) | Number of Total Leaves | Number of Green Leaves | Root Dry Weight (g/plant) | Leaf Dry Weight (g/plant) | Total Dry Weight (g/plant) | SPAD | Leaf Area (cm2) | Leaf Thickness (mm) | Net Photosynthetic Rate (µmol m⁻2 s⁻1) | Stomatal Conductance (µmol m⁻2 s⁻1) |
---|---|---|---|---|---|---|---|---|---|---|---|
CK | 7.16 b | 6.50 a | 6.33 a | 1.38 b | 12.41 b | 13.79 b | 43.47 b | 461.38 c | 0.517 a | 29.67 a | 0.91 a |
M | 7.65 ab | 6.50 a | 6.33 a | 1.48 ab | 13.52 b | 15.00 b | 44.90 a | 501.38 b | 0.518 a | 29.84 a | 0.93 a |
D | 8.06 a | 6.83 a | 6.67 a | 1.92 a | 16.86 a | 18.78 a | 44.95 a | 582.31 a | 0.519 a | 30.78 a | 0.97 a |
Treatment | Plant Height (cm) | Root Collar Diameter (mm) | Stem Diameter (cm) | Branch Position (cm) | Length of Main Inflorescence (cm) | Branch Number per Plant | Pods Number on the Main Stem | Branch Pods Number per Plant | Pods Number per Plant | Seeds Number per Pod | Seed Weight (g/1000) | Yield per Plant (g) | Dry Weight above Ground per Plant (g) |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
CK | 132.95 b | 1.64 a | 1.36 a | 30.70 b | 58.86 b | 8.14 a | 66.83 c | 183.83 a | 250.67 b | 25.97 a | 2.61 a | 16.94 b | 54.86 b |
M | 139.00 a | 1.66 a | 1.44 a | 36.30 a | 62.39 b | 8.00 a | 73.67 b | 203.17 a | 276.83 ab | 27.47 a | 2.61 a | 19.57 a | 62.90 a |
D | 140.90 a | 1.65 a | 1.44 a | 32.89 ab | 69.87 a | 8.00 a | 81.33 a | 212.67 a | 294.00 a | 28.34 a | 2.62 a | 21.50 a | 67.48 a |
Treatment | Protein Content (%) | Oil Content (%) | Glucosinolate (μmol/g) | Linolenic Acid (%) | Linoleic Acid (%) | Oleic Acid (%) |
---|---|---|---|---|---|---|
CK | 16.18 a | 46.91 a | 33.22 a | 7.03 a | 16.26 a | 69.49 a |
M | 15.87 a | 47.22 a | 31.00 b | 6.98 a | 16.26 a | 69.93 a |
D | 15.59 a | 47.54 a | 30.57 b | 6.66 a | 16.91 a | 69.60 a |
3. | Chlorophyll Content (mg/g) | Soluble Protein (g prot/L) | Proline (μg/g) | Malondialdehyde (μmol/g) | Soluble Sugar (mg/g) | SOD (U/mg prot) | POD (U/mg prot) | CAT (U/mg prot) |
---|---|---|---|---|---|---|---|---|
CK | 0.73 b | 10.65 b | 69.90 b | 0.30 a | 12.91 b | 13.10 b | 8.00 a | 7.58 a |
M | 1.02 a | 11.16 ab | 98.24 a | 0.22 ab | 17.96 a | 15.04 a | 7.00 a | 7.54 a |
D | 0.89 a | 12.17 a | 88.09 ab | 0.14 b | 16.16 a | 7.03 c | 6.33 a | 6.99 a |
KEGG Pathway | Genes | Metabolite |
---|---|---|
Phenylpropanoid biosynthesis | BnaC01G0012800ZS, novel.18876, BnaA10G0034100ZS | L-Phenylalanine, Spermidine, Sinapate, Ferulate, trans-2-Hydroxycinnamate |
2-Oxocarboxylic acid metabolism | BnaC02G0073500ZS, BnaA10G0179400ZS, BnaA04G0241000ZS | Indolylmethylthiohydroximate, 2-Oxoglutarate, L-Phenylalanine, Citrate, 4-Methylthiobutanaldoxime |
Citrate cycle (TCA cycle) | BnaA06G0006700ZS, BnaA08G0287200ZS, BnaC04G0228700ZS, BnaC02G0554900ZS | 2-Oxoglutarate, Citrate |
Alanine, aspartate, and glutamate metabolism | BnaA10G0179400ZS | 2-Oxoglutarate, Citrate |
Biosynthesis of amino acids | BnaA10G0179400ZS, BnaA03G0384700ZS, BnaC04G0009200ZS, BnaC02G0073500ZS, BnaA04G0241000ZS, BnaC04G0313900ZS, BnaC09G0538200ZS | 2-Oxoglutarate, L-Phenylalanine, Citrate |
Phenylalanine metabolism | BnaA10G0179400ZS | L-Phenylalanine, trans-2-Hydroxycinnamate |
Glyoxylate and dicarboxylate metabolism | BnaC01G0228200ZS, BnaC01G0037600ZS, BnaA01G0032600ZS | 2-Oxoglutarate, Citrate |
Carbon metabolism | BnaA10G0179400ZS, BnaC09G0057900ZS, BnaC02G0554900ZS, BnaC01G0037600ZS, BnaA01G0032600ZS, BnaA06G0006700ZS, BnaC04G0228700ZS | 2-Oxoglutarate, Citrate |
Arginine biosynthesis | BnaA10G0179400ZS, BnaA04G0241000ZS | 2-Oxoglutarate |
C5-Branched dibasic acid metabolism | BnaC02G0073500ZS | 2-Oxoglutarate |
Phenylalanine, tyrosine, and tryptophan biosynthesis | BnaA10G0179400ZS, BnaA03G0384700ZS | L-Phenylalanine |
Lysine biosynthesis | BnaC09G0538200ZS | 2-Oxoglutarate |
Glutathione metabolism | BnaA09G0015300ZS | Spermidine |
Butanoate metabolism | BnaC02G0073500ZS | 2-Oxoglutarate |
Cyanoamino acid metabolism | novel.18876 | L-Phenylalanine |
Ascorbate and aldarate metabolism | BnaC02G0069000ZS | 2-Oxoglutarate |
Aminoacyl-tRNA biosynthesis | BnaC08G0411300ZS, BnaA09G0059500ZS, BnaA04G0128500ZS | L-Phenylalanine |
Pentose and glucuronate interconversions | BnaA01G0353900ZS, BnaA07G0072600ZS, BnaC01G0473100ZS, Bnascaffold0027G0053200ZS, BnaC02G0069000ZS, BnaA05G0485000ZS | 2-Oxoglutarate |
Tropane, piperidine and pyridine alkaloid biosynthesis | BnaC03G0271400ZS, BnaA10G0179400ZS | L-Phenylalanine |
Flavonoid biosynthesis | BnaC02G0537300ZS | Kaempferol |
Tyrosine metabolism | BnaA04G0155600ZS, BnaA10G0179400ZS | 2-Hydroxy-3-(4-hydroxyphenyl)propenoate |
Arginine and proline metabolism | BnaA10G0179400ZS, BnaC09G0569400ZS, BnaC04G0313900ZS | Spermidine |
Tryptophan metabolism | novel.24812, BnaA01G0142600ZS, novel.24814, BnaA01G0032600ZS, BnaC01G0037600ZS | Indolylmethylthiohydroximate |
KEGG Pathway | Genes | Metabolite |
---|---|---|
Phenylpropanoid biosynthesis | BnaA09G0566300ZS, BnaC04G0553100ZS, BnaC03G0021400ZS, BnaC06G0428100ZS, BnaC06G0181900ZS, BnaA04G0299600ZS | L-Phenylalanine, Sinapate, 4-Coumarate, Ferulate, 5-Hydroxyferulic acid methyl este, Coumarin |
2-Oxocarboxylic acid metabolism | BnaA08G0059700ZS, BnaA01G0008700ZS, BnaA04G0241000ZS | L-Aspartate, L-Phenylalanine, L-Valine, Indolylmethylthiohydroximate, 4-Methylthiobutanaldoxime |
Glucosinolate biosynthesis | BnaA01G0008700ZS, BnaA08G0059700ZS | L-Phenylalanine, L-Valine, Indolylmethylthiohydroximate, 4-Methylthiobutanaldoxime |
Cyanoamino acid metabolism | BnaA09G0566300ZS, BnaA04G0299600ZS, BnaC03G0021400ZS | L-Aspartate, L-Phenylalanine, L-Valine |
Aminoacyl-tRNA biosynthesis | novel.636 | L-Aspartate, L-Phenylalanine, L-Valine |
Nicotinate and nicotinamide metabolism | BnaA02G0083500ZS, BnaC02G0270800ZS | L-Aspartate, Nicotinamide, N-Methylnicotinate |
Biosynthesis of amino acids | BnaC04G0009200ZS, BnaC06G0249000ZS, BnaA08G0001500ZS, BnaC03G0816400ZS, BnaA04G0241000ZS, BnaC02G0319900ZS | L-Aspartate, L-Phenylalanine, L-Valine |
Cysteine and methionine metabolism | BnaA03G0377000ZS, BnaC03G0461700ZS, BnaC03G0816400ZS, BnaA08G0001500ZS, BnaC04G0009200ZS, novel.50243, BnaC07G0293000ZS | L-Aspartate, Dehydroalanine |
Carbon fixation in photosynthetic organisms | BnaC06G0249000ZS, BnaA03G0427900ZS, BnaC02G0319900ZS, BnaC02G0209200ZS, BnaC09G0325700ZS | L-Aspartate |
Arginine biosynthesis | BnaA04G0241000ZS | L-Aspartate |
Sphingolipid metabolism | BnaA04G0244600ZS | Phytosphingosine |
Cutin, suberine, and wax biosynthesis | novel.25263 | Hexadecanoic acid |
Beta-Alanine metabolism | BnaC08G0024500ZS | L-Aspartate |
Valine, leucine, and isoleucine degradation | BnaC07G0327100ZS, BnaC08G0024500ZS, BnaC02G0488700ZS | L-Valine |
Fatty acid degradation | BnaA05G0470300ZS, BnaA05G0181300ZS, BnaC07G0327100ZS, BnaA09G0597200ZS, BnaC02G0488700ZS | Hexadecanoic acid |
Fatty acid biosynthesis | BnaA05G0470300ZS, BnaA05G0181300ZS, BnaA02G0298900ZS | Hexadecanoic acid |
Phenylalanine metabolism | BnaC04G0553100ZS, BnaC06G0181900ZS | L-Phenylalanine |
Tropane, piperidine, and pyridine alkaloid biosynthesis | novel.8259, BnaA05G0141600ZS | L-Phenylalanine |
Biosynthesis of unsaturated fatty acids | BnaC02G0488700ZS | Hexadecanoic acid |
Tyrosine metabolism | BnaA09G0597200ZS | 4-Coumarate |
Tryptophan metabolism | BnaA08G0059700ZS, BnaA01G0008700ZS, BnaA01G0142600ZS, novel.8529, BnaC07G0327100ZS | Indolylmethylthiohydroximate |
Purine metabolism | BnaC09G0106200ZS, BnaA01G0189300ZS, BnaA01G0356800ZS, BnaC02G0270800ZS, BnaC08G0375100ZS, BnaA09G0434400ZS, BnaC04G0298300ZS, BnaC02G0212200ZS | Adenosine |
Carbon metabolism | novel.18384, BnaA03G0427900ZS, BnaC03G0816400ZS, BnaC06G0249000ZS, BnaA08G0001500ZS, BnaC09G0325700ZS, BnaC02G0209200ZS, BnaC02G0319900ZS, BnaC07G0327100ZS, BnaC08G0024500ZS | L-Aspartate |
Fatty acid metabolism | BnaA05G0470300ZS, BnaC02G0488700ZS, BnaA05G0181300ZS, BnaC07G0327100ZS, BnaA02G0298900ZS | Hexadecanoic acid |
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Zhu, K.; Liu, J.; Lyu, A.; Luo, T.; Chen, X.; Peng, L.; Hu, L. Analysis of the Mechanism of Wood Vinegar and Butyrolactone Promoting Rapeseed Growth and Improving Low-Temperature Stress Resistance Based on Transcriptome and Metabolomics. Int. J. Mol. Sci. 2024, 25, 9757. https://doi.org/10.3390/ijms25179757
Zhu K, Liu J, Lyu A, Luo T, Chen X, Peng L, Hu L. Analysis of the Mechanism of Wood Vinegar and Butyrolactone Promoting Rapeseed Growth and Improving Low-Temperature Stress Resistance Based on Transcriptome and Metabolomics. International Journal of Molecular Sciences. 2024; 25(17):9757. https://doi.org/10.3390/ijms25179757
Chicago/Turabian StyleZhu, Kunmiao, Jun Liu, Ang Lyu, Tao Luo, Xin Chen, Lijun Peng, and Liyong Hu. 2024. "Analysis of the Mechanism of Wood Vinegar and Butyrolactone Promoting Rapeseed Growth and Improving Low-Temperature Stress Resistance Based on Transcriptome and Metabolomics" International Journal of Molecular Sciences 25, no. 17: 9757. https://doi.org/10.3390/ijms25179757