Reference Genes Selection of Gymnosporangium yamadae during the Interaction with Apple Leaves
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Material and Artificial Inoculation
2.2. RNA Extraction and cDNA Synthesis
2.3. Reference Gene Selection, Primer Design and RT-qPCR Parameters
2.4. Data Analysis
3. Results
3.1. Primer Specificity and Efficiency Evaluation
3.2. Expression Levels of Candidate Reference Genes in Different Samples
3.3. Stability of Candidate Reference Genes
3.4. Validation of Candidate Reference Genes
4. Discussion
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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Gene | Description | KOG ID | Sequence (5′-3′) | Amplicon Size (bp) | Efficiency (%) | R2 |
---|---|---|---|---|---|---|
ACT a | Actin and related proteins | KOG0676 | GACCATGTACTCGGGCATCTCAGCGAAGCCAGAATAGACC | 139 | - | - |
EF1 | Translation elongation factor EF-1 alpha | KOG0052 | AGGAGGCTCAATAGCGTCAA | 97 | 89.30 | 0.9970 |
CAACATGCAATGGTTCAAGG | ||||||
EF2 | Elongation factor 2 | KOG0469 | AGAAGCGAGGTCACGTGTTT | 155 | 98.40 | 0.9960 |
GTGGTCGAACACCATCTGTG | ||||||
GAPDH | Glyceraldehyde 3-phosphate dehydrogenase | KOG0657 | TCCTGCCTTTGAAATTTTGG | 101 | 114.50 | 0.9910 |
TTGCTTTACGCTTGATGTGC | ||||||
40S | 40S ribosomal protein S7 | KOG3320 | GGATCCATCGGTGTGAAATC | 104 | 91.16 | 0.9976 |
CTCGGTCACGAACACTCAAA | ||||||
60S a | 60S ribosomal protein L14/L17/L23 | KOG0901 | ATAGACGCGTTTTCGCTTGT | 195 | - | - |
CGGTAAAAGGCTCGTCTCAG | ||||||
β-TUB a | Beta tubulin | KOG1668 | CCGATCAATTCACAGCAATG | 195 | - | - |
CAGGGGGTACCTCATCTTCA | ||||||
α-TUB | Alpha tubulin | KOG1376 | ATATTTCCCGGAGCCAGTCT | 106 | 95.50 | 0.9950 |
TACCATCGAGCATGGTTTGA | ||||||
UBCE3 | E3 ubiquitin-protein ligase | KOG0939 | CCTTTGCTGGACTTTGAAGC | 183 | 103.10 | 0.9910 |
GCCTACACCAGAGGAGCTTG | ||||||
Cluster-3395.48660 b | - | - | GGAAGTGGCACCAACAAAGT | 84 | - | - |
ATCCACGACATTCGCATACA |
Parameters | Gene | |||||
---|---|---|---|---|---|---|
EF1 | EF2 | GAPDH | α-TUB | UBCE3 | 40S | |
Geo Mean | 18.35 | 20.53 | 19.58 | 20.79 | 22.83 | 21.15 |
AR Mean | 18.37 | 20.55 | 19.61 | 20.81 | 22.87 | 21.17 |
Min | 17.31 | 19.25 | 18.11 | 19.57 | 20.94 | 19.74 |
Max | 19.90 | 21.81 | 21.85 | 22.56 | 25.22 | 22.69 |
SD | 0.77 | 0.71 | 0.86 | 0.86 | 0.98 | 0.55 |
CV (%) | 4.21 | 3.43 | 4.38 | 4.12 | 4.30 | 2.61 |
x-fold Min [x-fold] | −2.04 | −2.43 | −2.76 | −2.33 | −3.71 | −2.66 |
x-fold Max [x-fold] | 2.94 | 2.43 | 4.85 | 3.43 | 5.22 | 2.89 |
x-fold SD [± x-fold] | 1.71 | 1.63 | 1.81 | 1.81 | 1.98 | 1.47 |
CC [r] | 0.991 | 0.915 | 0.930 | 0.938 | 0.892 | 0.768 |
p-value | 0.001 | 0.001 | 0.001 | 0.001 | 0.003 | 0.026 |
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Shao, C.; Lao, W.; Liang, Y. Reference Genes Selection of Gymnosporangium yamadae during the Interaction with Apple Leaves. J. Fungi 2022, 8, 830. https://doi.org/10.3390/jof8080830
Shao C, Lao W, Liang Y. Reference Genes Selection of Gymnosporangium yamadae during the Interaction with Apple Leaves. Journal of Fungi. 2022; 8(8):830. https://doi.org/10.3390/jof8080830
Chicago/Turabian StyleShao, Chenxi, Wenhao Lao, and Yingmei Liang. 2022. "Reference Genes Selection of Gymnosporangium yamadae during the Interaction with Apple Leaves" Journal of Fungi 8, no. 8: 830. https://doi.org/10.3390/jof8080830