Cytogenetic and Molecular Marker Analyses of a Novel Wheat–Psathyrostachys huashanica 7Ns Disomic Addition Line with Powdery Mildew Resistance
Abstract
:1. Introduction
2. Results
2.1. Chromosomal Composition of 18-1-5
2.2. Non-Denaturing Fluorescence In Situ Hybridization (ND-FISH) Analysis of 18-1-5
2.3. Molecular Marker Analysis of 18-1-5
2.4. Powdery Mildew Response of 18-1-5
2.5. Agronomic Performance of 18-1-5
2.6. Acquisition of Specific P. huashanica 7Ns Chromosome Sequences
2.7. Development and Validation of Specific Molecular Markers
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Sequential GISH and Mc-FISH Analyses
4.3. ND-FISH Analysis
4.4. Molecular Marker Analysis
4.5. Powdery Mildew Response
4.6. Agronomic Trait Evaluation
4.7. Genotyping-by-Sequencing and Data Analysis
4.8. Development and Verification of PCR-Based Markers
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
CS | Chinese Spring |
CSph2b | Chinese Spring ph2b mutant |
mc-FISH | Multicolor-fluorescence in situ hybridization |
ND-FISH | Non-denaturing fluorescence in situ hybridization |
GISH | Genomic in situ hybridization |
PLUG | PCR-based landmark unique gene |
GBS | Genotyping-by-sequencing |
SLAF | Specific locus-amplified fragment sequencing |
PCR | Polymerase chain reaction |
DAPI | 4,6-Diamino-2-phenylindole |
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Lines | Plant Height (cm) | Tiller Number | Spike Length (cm) | Spikelets per Spike | Kernels per Spike | Kernel Length (mm) | Kernel Width (mm) | Thousand-Kernel Weight (g) |
---|---|---|---|---|---|---|---|---|
CS | 140.11 ± 1.28 Aa | 8.40 ± 0.51 Aa | 8.34 ± 0.13 Aa | 21.60 ± 0.31 Aa | 53.87 ± 1.32 Aa | 5.97 ± 0.03 Aa | 3.29 ± 0.02 Aa | 30.29 ± 0.26 Aa |
CSph2b | 133.88 ± 1.63 Bb | 7.93 ± 0.37 Aa | 8.70 ± 0.13 Aa | 22.20 ± 0.30 Aa | 55.80 ± 1.68 Aa | 6.07 ± 0.05 Aa | 3.21 ± 0.03 Ab | 29.58 ± 0.27 Aa |
18-1-5 | 111.87 ± 1.74 Cc | 7.47 ± 0.58 Aa | 8.67 ± 0.15 Aa | 20.00 ± 0.37 Bb | 46.47 ± 1.59 Bb | 6.84 ± 0.08 Bb | 2.91 ± 0.03 Bc | 27.79 ± 0.62 Bb |
Sample | Raw Base (bp) | Clean Base (bp) | Effective Rate (%) | Error Rate (%) | Q20 (%) | Q30 (%) | GC Content (%) |
---|---|---|---|---|---|---|---|
CSph2b | 1,404,766,080 | 1,404,693,792 | 99.99 | 0.05 | 94.21 | 85.77 | 43.67 |
ZY3156 | 1,809,454,176 | 1,809,426,816 | 100.00 | 0.05 | 94.35 | 85.76 | 44.20 |
18-1-5 | 2,734,574,112 | 2,734,299,360 | 99.99 | 0.04 | 96.87 | 91.07 | 44.55 |
Sample | Total Reads | Unmapped Reads | uniqReads |
---|---|---|---|
CSph2b | 9,754,818 | 47,036 | |
ZY3156 | 12,565,464 | 6,476,616 | |
18-1-5 | 18,988,190 | 429,274 | 15,925 |
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Tan, B.; Wang, M.; Cai, L.; Li, S.; Zhu, W.; Xu, L.; Wang, Y.; Zeng, J.; Fan, X.; Sha, L.; et al. Cytogenetic and Molecular Marker Analyses of a Novel Wheat–Psathyrostachys huashanica 7Ns Disomic Addition Line with Powdery Mildew Resistance. Int. J. Mol. Sci. 2022, 23, 10285. https://doi.org/10.3390/ijms231810285
Tan B, Wang M, Cai L, Li S, Zhu W, Xu L, Wang Y, Zeng J, Fan X, Sha L, et al. Cytogenetic and Molecular Marker Analyses of a Novel Wheat–Psathyrostachys huashanica 7Ns Disomic Addition Line with Powdery Mildew Resistance. International Journal of Molecular Sciences. 2022; 23(18):10285. https://doi.org/10.3390/ijms231810285
Chicago/Turabian StyleTan, Binwen, Miaomiao Wang, Li Cai, Sanyue Li, Wei Zhu, Lili Xu, Yi Wang, Jian Zeng, Xing Fan, Lina Sha, and et al. 2022. "Cytogenetic and Molecular Marker Analyses of a Novel Wheat–Psathyrostachys huashanica 7Ns Disomic Addition Line with Powdery Mildew Resistance" International Journal of Molecular Sciences 23, no. 18: 10285. https://doi.org/10.3390/ijms231810285