Screening and Identification of Garlic Leaf Blight (Pleospora herbarum)-Resistant Mutants Induced by Ethyl Methane Sulphonate
Abstract
:1. Introduction
2. Results
2.1. Selection of EMS-Induced Mutant with Garlic G024
2.2. Variation of Agronomic Traits in M2 Mutants
2.3. Variation of Agronomic Traits in M3 Mutants
2.4. Screening and Identification of Mutants Resistant to Leaf Blight
2.5. Differentially Expressed Genes between A150 and G024 after Pleospora herbarum Infection
2.6. Differentially Expressed Genes Are Distributed in Different Functional Pathways
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Establishment of EMS Mutation Technology
4.3. Field Experiments and Plant Growth
4.4. Screening and Identification of EMS Mutated Lines Resistant to Leaf Blight
4.4.1. Phenotypic Observations
4.4.2. Fungal Inoculation
4.5. Transcriptome Analysis and Functional Annotation
4.6. Validation of RNA-Seq Data
4.7. Data Collection and Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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EMS Concentration (%) | EMS Treatment Time (h) | No. of Cloves | No. of Explants for Germination | Germination Rate (%) |
---|---|---|---|---|
0% | 0 | 50 | 49.6 ± 0.33 | 99.33 ± 1.15 a |
1.00% | 4 | 50 | 35 ± 0.57 | 70.00 ± 2.00 b |
8 | 50 | 24.6 ± 0.88 | 49.33 ± 3.05 c | |
12 | 50 | 16.6 ± 0.88 | 32.66 ± 3.05 e | |
16 | 50 | 10 ± 1.15 | 20.00 ± 4.00 f | |
24 | 50 | 3 ± 1.00 | 6.00 ± 3.46 h | |
1.50% | 4 | 50 | 26.6 ± 1.20 | 51.33 ± 4.16 c |
8 | 50 | 21.6 ± 1.45 | 43.33 ± 5.03 d | |
12 | 50 | 11.3 ± 0.88 | 22.66 ± 3.05 f | |
16 | 50 | 6.3 ± 0.88 | 12.66 ± 3.05 g | |
24 | 50 | 0.6 ± 0.33 | 1.33 ± 1.15 h |
Mutant Organ | Mutation Characteristics | Number of M1 Mutations | M1 Mutation Frequency |
---|---|---|---|
Blade | Albinism | 1 | 0.29‰ |
Mosaicism | 3 | 0.88‰ | |
Curly leaves | 2 | 0.58‰ | |
Involucre | Expansion | 6 | 1.7‰ |
Tiller | Tiller abnormality | 3 | 0.88‰ |
Plant | Dwarfism | 1 | 0.29‰ |
Robustness | 161 | 4.80% | |
Disease resistance | 17 | 0.53% | |
Total | 199 | 6.20% |
Trait | G024 | Mean of Mutants | Range | CV |
---|---|---|---|---|
Leaf length (cm) | 43.53 ± 0.13 | 54.18 ± 5.4 | 43.53–66.33 | 0.12 |
Leaf width (mm) | 24.38 ± 0.17 | 29.68 ± 3.98 | 21.44–36 | 0.11 |
Pseudo-stem height (cm) | 37.17 ± 0.15 | 36.62 ± 2.82 | 27.67–43.33 | 0.01 |
Stem diameter (mm) | 13.1 ± 0.22 | 16.31 ± 5.19 | 11.67–21.46 | 0.12 |
Leaf number | 7.7 ± 0.12 | 7.85 ± 0.86 | 6.67–9.33 | 0.01 |
Garlic weight (g) | 23.1 ± 0.12 | 34.78 ± 4.65 | 13.97–55.19 | 0.12 |
Garlic height (mm) | 29.77 ± 0.02 | 33.23 ± 2.66 | 24.04–39.85 | 0.06 |
Garlic diameter (mm) | 40.09 ± 0.09 | 47.23 ± 3.19 | 32.63–56.53 | 0.09 |
Petal number | 10 ± 0.1 | 12.77 ± 8.22 | 7–17.67 | 0.13 |
Trait | G024 | Mean of Mutants | Range | CV |
---|---|---|---|---|
Leaf length (cm) | 42.67 ± 1.20 | 54.80 ± 0.29 | 41.67–63.67 | 0.14 |
Leaf width (mm) | 27.17 ± 1.08 | 25.25 ± 0.14 | 21.25–34.34 | 0.04 |
Pseudo-stem height (cm) | 42.33 ± 1.67 | 46.12 ± 0.28 | 36.67–56.67 | 0.05 |
Stem diameter (mm) | 15.93 ± 1.01 | 13.64 ± 0.11 | 11.06–18.05 | 0.08 |
Leaf number | 6.67 ± 0.33 | 6.83 ± 0.03 | 6.00–8.33 | 0.01 |
Garlic weight (g) | 29.41 ± 1.87 | 48.50 ± 0.59 | 18.83–63.22 | 0.27 |
Garlic height (mm) | 31.53 ± 0.82 | 33.08 ± 0.16 | 25.82–38.12 | 0.03 |
Garlic diameter (mm) | 44.81 ± 1.76 | 48.50 ± 0.26 | 39.30–56.00 | 0.04 |
Petal number | 12.33 ± 0.33 | 12.47 ± 0.09 | 9.67–15.67 | 0.006 |
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Hong, Y.; Sun, Y.; Zhang, X.; Zhang, L.; Yuan, X.; Ma, Z.; Wu, M.; Chen, S. Screening and Identification of Garlic Leaf Blight (Pleospora herbarum)-Resistant Mutants Induced by Ethyl Methane Sulphonate. Int. J. Mol. Sci. 2023, 24, 11819. https://doi.org/10.3390/ijms241411819
Hong Y, Sun Y, Zhang X, Zhang L, Yuan X, Ma Z, Wu M, Chen S. Screening and Identification of Garlic Leaf Blight (Pleospora herbarum)-Resistant Mutants Induced by Ethyl Methane Sulphonate. International Journal of Molecular Sciences. 2023; 24(14):11819. https://doi.org/10.3390/ijms241411819
Chicago/Turabian StyleHong, Yuanyuan, Yinhui Sun, Xuan Zhang, Lingling Zhang, Xin Yuan, Zhaoyang Ma, Meiqian Wu, and Shuxia Chen. 2023. "Screening and Identification of Garlic Leaf Blight (Pleospora herbarum)-Resistant Mutants Induced by Ethyl Methane Sulphonate" International Journal of Molecular Sciences 24, no. 14: 11819. https://doi.org/10.3390/ijms241411819