Molecular Cytogenetic Identification of the Wheat–Dasypyrum villosum T3DL·3V#3S Translocation Line with Resistance against Stripe Rust
Abstract
:1. Introduction
2. Results
2.1. Generation and Screening for Wheat–D. villosum 3V#3.3D Translocation Stocks
2.2. Identification of T. aestivum–D. villosum Structural Aberrations by ND-FISH
2.3. Molecular Marker Analysis
2.4. Stripe Rust Response
2.5. Development and Verification of 3V#3-Specific EST Markers
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. 60Co γ-Irradiation Treatment of CD-3 Seeds
4.3. Mitotic Analysis and Survival Rate Investigation of the M0 Generation
4.4. Statistics Analysis
4.5. FISH Analysis
4.6. Molecular Marker Analysis
4.7. Assessment of Adult Plant Resistance to Stripe Rust
4.8. RNA-Seq and Transcriptome Assembly
4.9. Development and Validation of 3V#3 Chromosome-Specific EST Markers
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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Marker Name | Primer Sequence (5′–3′) | Wheat Bin Map Location | Wheat Chromosomal Location | Restriction Enzymes | Length of 3V Bands, bp |
---|---|---|---|---|---|
TNAC1248 | F: ATGATGCAGCAGCAAATTACA | 3AS4-0.45-1.00 | 3AS-211.50 | TaqI | 1000 |
R: CTGAGGAGCCTCTCCAACTCT | C-3BS1-0.33 | 3BS-251.65 | |||
3DS3-0.24-0.31 | 3DS-172.90 | ||||
TNAC1326 | F:ACAGATCGAGATGTTTATTGAAA | 3AS4-0.45-1.00 | 3AS-137.91 | TaqI | 750 |
R: GATCAAAGAGATGCGCTGAAG | 3BS1-0.33-0.57 | 3BS-181.49 | |||
3DS10-0.31-0.44 | 3DS-127.34 | ||||
TNAC1267 | F: GAGAGGCAGCTTCACTAGCAG | 3AL3-0.42-0.61 | 3AL-522.17 | TaqI | 200 |
R: CGTCAGGATCAGCTCTCATGT | 3BL2-0.22-0.41 | 3BL-527.29 | |||
3DL1-0.23-0.81 | 3DL-401.87 | ||||
TNAC1359 | F: GTAAATAGCGCCATCTGCGTA | 3AL3-0.42-0.61 | 3AL-531.08 | TaqI | 750 |
R: CTCTGGATGCAGTTGGAATGT | 3BL3-0.41-0.50 | 3BL-547.12 | |||
3DL1-0.23-0.81 | 3DL-419.90 |
Primers No. | Primer Sequences(5’–3’) | Localization in D. villosum#3 | Localization in Wheat | Product Size in D. villosum#3 (bp) | Product Size in D. villosum#4 (bp) | |
---|---|---|---|---|---|---|
Forward | Reverse | |||||
3V#3-1 | GCCTCATGCGGCTGTTGG | GAGGTCATGGTGAGCACGAGA | 3V#3S | 3BS | 148 | - |
3V#3-2 | CGGCAAGAGGTCGATGGT | TGACCCACGCACGCACTA | 3V#3L | 1DS | 210 | - |
3V#3-3 | GCGTCTTGGATGTCCTG | CGATTTGCTGCCCTACAT | 3V#3L | 3AL | 177 | - |
3V#3-4 | GCATGATAGAGAGGTTAGCCAT | CACTGGTGATGTTGTTCAGTACT | 3V#3S | 1AL | 90 | - |
3V#3-5 | TCGATACAATTGTTCTTGAGATATG | ACTGGTGCCCTCTTGACG | 3V#3L | 3AL | 560 | - |
3V#3-6 | AAACAATCTAGCACTACCCAGAGG | AAGAGGAAGAGAAATAAGCGAGG | 3V#3L | 3DL | 166 | - |
3V#3-7 | AGGTCCTTGTCCGAGGTGAT | ATGTTACCGATACTGATGCCACT | 3V#3S | 3AS | 982 | - |
3V#3-8 | ATGCTGAACGCAAGGTCAAATA | TGCTGAAGCCCATCACGAAG | 3V#3L | 3DL | 1500 | - |
3V#3-9 | CGACTGGTCCACCGTTTC | CGCTGCCTAGTTACCTCTGTT | 3V#3L | 3DL | 1700 | - |
3V#3-10 | CTTTCAAGGTAATCCCAGAACT | TGGAAAGCAAACAGGATACG | 3V#3S | 3AS | 126 | - |
3V#3-11 | TACGAATAACAACTGCAAGCAGAAT | TGCTAATGGCATCAGCGTCA | 3V#3L | 3DL | 1302 | - |
3V#3-12 | CGACTGGTCCACCGTTTC | CGCTGCCTAGTTACCTCTGTT | 3V#3L | 3AL | 1700 | - |
3V#4-1 * | TCCATCATAGCACCTTCAGACTCAAG | GACAACTCGGCAATCACCAAGGA | 3V#3S | 5BL | 850 | 377 |
3V#4-2 * | GGCAACTCAAATTATAGGATCACGAC | GCAAGGCGGAGTAGCTCACA | 3V#3L | 5DL | 243 | 243 |
3V#4-3 * | TTCGTCATCTTTGTTGACATGGCAA | GCAAGGCGGAGTAGCTCACA | 3V#3L | 5DL | 240 | 263 |
3V#4-5 * | TTAGAGCGACGACAACTATGC | CAGTATAAGATAGGAGAAGCGACAG | 3V#3L | 3AL | 175 | 175 |
3V#4-7 * | GTTATATCGGTTGAGGCGTCTATAC | AACAGTGAGTTCTTCAGGACAGA | 3V#3L | 3DL | 700 | 341 |
3V#4-11 * | CTCGTCGGTCTCAGAAGTCAA | TCCACAGAATCATCGGCTCTC | 3V#3L | 3DL | 760 | 760 |
3V#4-12 * | CCTCCTCTTCCTCCTCTTCC | TCGCACCATCACCGTACTT | 3V#3L | 3DL | 499 | 499 |
3V#4-14 * | GGACGGATGTAGTCTTGTTCAA | CTCGTATCGTACTGCTACTCA | 3V#3L | 3DL | 593 | 593 |
3V#4-16 * | GTCCACCAAATCACATCAAACA | GCTCTCACAAGTCACAACAATT | 3V#3L | 3DL | 180 | 180 |
3V#4-17 * | ACCATATACTTCGGTGGAACATAC | GCATAGTTACTCTATCACAGACTCA | 3V#3L | 3DL | 304 | 304 |
3V#4-18 * | TACCATATACTTCGGTGGAACATAC | GCATAGTTACTCTATCACAGACTCA | 3V#3L | 3DL | 305 | 305 |
3V#4-19 * | TGCTCTTCACAGTTCATCTCCT | AGACAAGTTCAGTTCCACACTC | 3V#3L | 3DL | 364 | 364 |
3V#4-20 * | TGGTTGCTTCTCAGTTGTGTTG | TACTCGGATAGTGCCTTGTTGA | 3V#3S | 3AS | 600 | 237 |
3V#4-21 * | GTTGCTTCTCAGTTGTGTTGGA | TACTCGGATAGTGCCTTGTTGA | 3V#3S | 3AS | 600 | 235 |
3V#4-22 * | TGGTTGCTTCTCAGTTGTGTTG | CGGATAGTGCCTTGTTGATGAC | 3V#3S | 3AS | 600 | 233 |
3V#4-24 * | GAGAACTGCTCAACATGACAATAAG | CAACAGTATCATCAATGGAGGTCTT | 3V#3S | 3DS | 144 | 144 |
3V#4-26 * | TCGCCAGCACCAACCAAT | CAGCACAGCACACCAATGAA | 3V#3S | 3DS | 686 | 686 |
3V#4-27 * | GTGACACCAATAGAAGGCAGAA | GGAGGAGCATACCGTGGAA | 3V#3S | 3AS | 403 | 403 |
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Zhang, J.; Tang, S.; Lang, T.; Wang, Y.; Long, H.; Deng, G.; Chen, Q.; Guo, Y.; Xuan, P.; Xiao, J.; et al. Molecular Cytogenetic Identification of the Wheat–Dasypyrum villosum T3DL·3V#3S Translocation Line with Resistance against Stripe Rust. Plants 2022, 11, 1329. https://doi.org/10.3390/plants11101329
Zhang J, Tang S, Lang T, Wang Y, Long H, Deng G, Chen Q, Guo Y, Xuan P, Xiao J, et al. Molecular Cytogenetic Identification of the Wheat–Dasypyrum villosum T3DL·3V#3S Translocation Line with Resistance against Stripe Rust. Plants. 2022; 11(10):1329. https://doi.org/10.3390/plants11101329
Chicago/Turabian StyleZhang, Jie, Shuyao Tang, Tao Lang, Ying Wang, Hai Long, Guangbing Deng, Qian Chen, Yuanlin Guo, Pu Xuan, Jun Xiao, and et al. 2022. "Molecular Cytogenetic Identification of the Wheat–Dasypyrum villosum T3DL·3V#3S Translocation Line with Resistance against Stripe Rust" Plants 11, no. 10: 1329. https://doi.org/10.3390/plants11101329