Transcriptomics Integrated with Changes in Cell Wall Material of Chestnut (Castanea mollissima Blume) during Storage Provides a New Insight into the “Calcification” Process
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Materials and Treatments
2.2. Determination of the Moisture Content
2.3. Determination of Cell Wall Material in Different Calcified Chestnut Seeds
2.3.1. Extraction of the Cell Wall Material
2.3.2. Extraction and Determination of the Lignin and Cellulose Contents
2.3.3. Extraction and Determination of Pectin Content with Different Solubilities
2.4. Transcriptome Analysis of Chestnut with Different Degrees of Calcification
2.4.1. RNA Extraction and Transcriptome Sequencing
2.4.2. Analysis of Differentially Expressed Genes
2.4.3. Gene Set Enrichment Analysis
2.4.4. Quantitative Real-Time PCR (qRT-PCR) Analysis
2.4.5. Statistical Analyses
3. Results
3.1. Relationship between the Degree of Chestnut Calcification and Moisture Content
3.2. Changes in Cell Wall Material (CWM) Content in Different Stages of Chestnut Calcification
3.3. Changes in the Lignin and Cellulose Content of Different Stages of Calcification
3.4. Changes in the Pectin Content of Chestnut at Different Stages of Calcification
3.5. Transcriptome Sequencing and Assembly
3.6. Comparisons between the 0% and 50% Stages of Calcification
3.7. GO and KEGG Enrichment of the DEGs
3.8. GSEA Enrichment Analysis
3.9. Transcriptomic Insights into Chestnut Calcification and Moisture Content
3.10. Transcriptomic Insights into Cellulose and Lignin Metabolism
3.11. Pectin Metabolism-Related DEGs
3.12. Validation of Transcriptome Data by qRT-PCR
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
References
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Samples | Clean Reads | Clean Data | GC content (%) | ≥Q30 |
---|---|---|---|---|
T1a | 32,619,737 b | 8,154,934,250 b | 44.91 | 93.50 |
T1b | 30,425,950 d | 7,606,487,500 d | 44.95 | 93.57 |
T1c | 23,793,560 e | 5,948,390,000 e | 45.06 | 93.13 |
T3a | 31,853,069 c | 7,963,267,250 c | 45.22 | 93.73 |
T3b | 34,018,941 a | 8,504,735,250 a | 45.29 | 93.36 |
T3c | 32,735,082 b | 8,183,770,500 b | 45.06 | 93.36 |
Samples | Total Reads | Mapped Reads | Unique Mapped | Multiple Mapped |
---|---|---|---|---|
T1a | 65,239,474 b | 54,516,406 c | 53,664,614 c | 851,792 b |
T1b | 60,851,900 d | 50,891,629 d | 49,885,627 d | 1,006,002 a |
T1c | 47,587,120 e | 39,889,854 e | 39,123,856 e | 765,998 c |
T3a | 63,706,138 c | 54,636,733 c | 53,975,213 c | 661,520 d |
T3b | 68,037,882 a | 58,347,962 a | 57,572,054 a | 775,908 c |
T3c | 65,470,164 b | 56,715,906 b | 56,031,375 b | 684,531 d |
Gene | FPKM (0%) | FPKM (50%) | FDR | Log2FC | Regulated |
---|---|---|---|---|---|
Cellulose biosynthetic process | |||||
Scaffold01281-gene-0.39 | 7.13 | 1.59 | 0.0034 | −1.84 | Down |
Scaffold01202-gene-0.30 | 112.48 | 42.79 | 0.00012 | −2.00 | Down |
Scaffold03287-gene-0.16 | 2.05 | 0.43 | 0.00100 | −4.46 | Down |
Scaffold00980-gene-0.41 | 0.08 | 0.03 | 0.00681 | −1.51 | Down |
Scaffold02563-gene-0.4 | 33.76 | 10.17 | 0.00549 | −1.95 | Down |
Scaffold11300-gene-0.9 | 4.83 | 16.05 | 0.00676 | 1.73 | Up |
Sucrose synthase activity | |||||
Scaffold0125-gene-0.31 | 78.45 | 30.52 | 0.00358 | −1.44 | Down |
Lignin catabolic process | |||||
Scaffold34956-gene-0.3 | 8.24 | 1.04 | 0.00128 | −3.09 | Down |
Scaffold08019-gene-0.16 | 0.71 | 0.05 | 0.00021 | −4.35 | Down |
Gene | FPKM (0%) | FPKM (50%) | FDR | Log2FC | Regulated |
---|---|---|---|---|---|
Pectin biosynthetic process | |||||
Scaffold02462-gene-0.13 | 1.15 | 0.13 | 1.34 × 10−7 | −3.60 | Down |
Scaffold00897-gene-0.33 | 20.93 | 7.75 | 0.00201 | −1.53 | Down |
Scaffold02056-gene-0.28 | 2.7 | 0.28 | 0.00554 | −2.74 | Down |
Scaffold04120-gene-0.22 | 2.19 | 25.97 | 4.74 × 10−6 | 3.66 | Up |
Pectinesterase activity | |||||
Scaffold11321-gene-0.6 | 33.66 | 0.22 | 2.92 × 10−16 | −7.86 | Down |
Scaffold01256-gene-0.30 | 33.61 | 0.02 | 4.48 × 10−9 | −4.36 | Down |
Scaffold00325-gene-0.6 | 0.25 | 0.03 | 0.00563 | −4.85 | Down |
Polygalacturonase activity | |||||
Scaffold02071-gene-0.36 | 6.75 | 1.15 | 0.00016 | −2.43 | Down |
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Chen, Y.; Zhu, C.; Zhao, Y.; Zhang, S.; Wang, W. Transcriptomics Integrated with Changes in Cell Wall Material of Chestnut (Castanea mollissima Blume) during Storage Provides a New Insight into the “Calcification” Process. Foods 2022, 11, 1136. https://doi.org/10.3390/foods11081136
Chen Y, Zhu C, Zhao Y, Zhang S, Wang W. Transcriptomics Integrated with Changes in Cell Wall Material of Chestnut (Castanea mollissima Blume) during Storage Provides a New Insight into the “Calcification” Process. Foods. 2022; 11(8):1136. https://doi.org/10.3390/foods11081136
Chicago/Turabian StyleChen, Yu, Cancan Zhu, Yuqiang Zhao, Shijie Zhang, and Wu Wang. 2022. "Transcriptomics Integrated with Changes in Cell Wall Material of Chestnut (Castanea mollissima Blume) during Storage Provides a New Insight into the “Calcification” Process" Foods 11, no. 8: 1136. https://doi.org/10.3390/foods11081136