Identification of Oil Palm’s Consistently Upregulated Genes during Early Infections of Ganoderma boninense via RNA-Seq Technology and Real-Time Quantitative PCR
Abstract
:1. Introduction
2. Results
2.1. Transcriptomic Analysis via RNA-Seq
2.2. Identification of DEGs
2.3. Functional Annotation
2.4. Gene Expression Profiling of Selected DEGs via qRT-PCR
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Treatments for Transcriptomic Profiling
4.2. Total RNA Extraction and cDNA Synthesis
4.3. RNA-Sequencing and Transcriptome Library Generation
4.4. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Samples | Total Reads (bp) | Low-Quality Reads (%) | Total High-Quality Reads (bp) | PhiX Contamination (%) | Total Sample Reads (bp) |
---|---|---|---|---|---|
REI | 16,536,956 | 8.51 | 15,129,373 | 0.32 | 15,080,896 |
RLI | 21,075,290 | 9.49 | 19,075,010 | 0.31 | 19,014,726 |
REC | 17,741,550 | 8.71 | 16,195,459 | 0.33 | 16,141,333 |
RLC | 27,247,240 | 10.79 | 24,305,602 | 0.32 | 24,227,629 |
Oil Palm Root Sample Pairs | Total Significant DEGs | Up-Regulated Transcripts | Down-Regulated Transcripts |
---|---|---|---|
REI/REC | 21 | 12 | 9 |
RLI/RLC | 103 | 69 | 34 |
Gene ID | Annotation Genes | Labels | FPKM | Log 2 Fold-Change | |
---|---|---|---|---|---|
Control | Treated | ||||
Highly Upregulated Root Genes at Early Stage of Infection (REI/REC) | |||||
XLOC_022484 | anthocyanidin synthase | ANTHO | 5.95 | 189.31 | 31.79 |
XLOC_009804 | leucoanthocyanidin reductase-like | LEUCO | 16.66 | 198.57 | 11.91 |
XLOC_019742 | ethylene-responsive transcription factor 1b-like | ETHYLENE | 4.09 | 109.77 | 26.85 |
XLOC_005112 | flavonoid 3–hydroxylase | FLAVONOID | 0 | 19.56 | Positive Infinity |
XLOC_013737 | mannose-specific lectin-like | MANNOSE | 85.08 | 1114.43 | 13.09 |
Highly Upregulated Root Genes at Late Stage of Infection (RLI/RLC) | |||||
XLOC_016957 | chalcone synthase | CHALCONE | 0 | 674.91 | Positive Infinity |
XLOC_009482 | glucan endo-beta-glucosidase-like | GLUCAN | 555.34 | 9543.37 | 17.18 |
XLOC_009990 | nadph:quinone oxidoreductase-like | NADPH | 5.69 | 107.89 | 18.96 |
XLOC_017303 | senescence-associated partial | SENESCENCE | 91.86 | 1540.91 | 16.77 |
XLOC_001542 | thaumatin-like protein | THAUMATIN | 13.10 | 187.77 | 14.33 |
Gene | Amplicon Size (bp) | BLAST Results | Score (Bits) | E-Value | Percentage of Identity (%) | Accession No. |
---|---|---|---|---|---|---|
ANTHO | 138 | Anthocyanidin synthase (Delphinium grandiflorum) | 51.6 | 9 × 10−7 | 92 | BAO04186.1 |
LEUCO | 170 | PREDICTED: leucoanthocyanidin reductase-like(Elaeis guineensis) | 72.4 | 4 × 10−14 | 100 | XP 010916146.1 |
ETHYLENE | 142 | PREDICTED: ethylene-responsive transcription factor 1B-like (Elaeis guineensis) | 70.5 | 2 × 10−13 | 100 | XP 010904582.1 |
FLAVONOID | 155 | flavonoid 3′–hydroxylase (Sorghum bicolor) | 56.2 | 3 × 10−8 | 83 | ABG54321.1 |
MANNOSE | 105 | PREDICTED: mannose-specific lectin-like(Elaeis guineensis) | 43.9 | 3 × 10−4 | 100 | XP 010910930.1 |
CHALCONE | 135 | PREDICTED: chalcone synthase-like (Elaeis guineensis) | 48.1 | 9 × 10−5 | 85 | XP 010910945.1 |
GLUCAN | 110 | PREDICTED: glucan endo- 1,3-beta-glucosidase-like (Elaeis guineensis) | 36.2 | 0.67 | 79 | XP 010921890.1 |
NADPH | 116 | PREDICTED: NADPH:quinone oxidoreductase-like (Elaeis guineensis) | 40.8 | 0.010 | 100 | XP 010922202.1 |
SENESCENCE | 169 | putative senescence-associated protein (Cupressus sempervirens) | 90.5 | 1 × 10−21 | 100 | ACA30301.1 |
THAUMATIN | 110 | PREDICTED: Elaeis guineensis thaumatin-like protein (LOC105033331), mRNA | 75.0 | 2 × 10−10 | 94 | XM 010908090.1 |
Primer Name | Primer Sequence (5′–3′) | Product Length (bp) | |
---|---|---|---|
Forward Primer | Reverse Primer | ||
ANTHO | ACAACATGGTCCCCGGTCT | GGTGGAGGATGCTCTTGTAGGT | 138 |
LEUCO | TCCGTTTTGGGCGGTTCT | CGGCGGACTTTCCTCTTTTC | 170 |
ETHYLENE | AAGAGCAAGGCAGGGAATGG | CTTCTGCGCTGTCAAAGGTTC | 142 |
FLAVONOID | GTTTGTGGTGGGAGACTTCGTG | CCTCATTCTGCTCGGTTGGAC | 155 |
MANNOSE | TCGGATGGGAACCTTGTGG | CCGATCTCGTTGGAGGATACAG | 105 |
CHALCONE | GAGCAGATCCAATGCAAGGTGT | GGTTGAGGAGGTGGAAGGTGA | 135 |
GLUCAN | AGCAAGCTACTGGGTCCAAAC | GCACATACTGGGCTTTATCTCC | 110 |
NADPH | CGAGATTGATGGCAAGTGTCC | TCAGAGGAGCTGGGATGGAGT | 116 |
SENESCENCE | GGCACGGCCATCAGTAGAGTA | AGCCAAGCGTTCATAGCGAC | 169 |
THAUMATIN | ACGAGGGAGATGTCGATGAA | GACTGCGGTGGTAAACTTGC | 110 |
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Zuhar, L.M.; Madihah, A.Z.; Ahmad, S.A.; Zainal, Z.; Idris, A.S.; Shaharuddin, N.A. Identification of Oil Palm’s Consistently Upregulated Genes during Early Infections of Ganoderma boninense via RNA-Seq Technology and Real-Time Quantitative PCR. Plants 2021, 10, 2026. https://doi.org/10.3390/plants10102026
Zuhar LM, Madihah AZ, Ahmad SA, Zainal Z, Idris AS, Shaharuddin NA. Identification of Oil Palm’s Consistently Upregulated Genes during Early Infections of Ganoderma boninense via RNA-Seq Technology and Real-Time Quantitative PCR. Plants. 2021; 10(10):2026. https://doi.org/10.3390/plants10102026
Chicago/Turabian StyleZuhar, Liyana Mohd, Ahmad Zairun Madihah, Siti Aqlima Ahmad, Zamri Zainal, Abu Seman Idris, and Noor Azmi Shaharuddin. 2021. "Identification of Oil Palm’s Consistently Upregulated Genes during Early Infections of Ganoderma boninense via RNA-Seq Technology and Real-Time Quantitative PCR" Plants 10, no. 10: 2026. https://doi.org/10.3390/plants10102026