Marker Development for Differentiation of Fusarium oxysporum f. sp. Niveum Race 3 from Races 1 and 2
Abstract
:1. Introduction
2. Results
2.1. Sequence Analysis and Primer Design
2.2. Specificity and Sensitivity of the Race 3 Marker
2.3. Development of a Protocol for Race Differentiation
2.4. Race Distribution of Experimental Samples
3. Discussion
4. Materials and Methods
4.1. FON Isolates and DNA Extraction
4.2. Sequence Alignment and Primer Design
4.3. Selection of Race-Specific Diagnostic Marker Primer Set and PCR Conditions
4.4. Optimization of the Developed Marker
4.5. Development and Application of a Protocol for Race Differentiation
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
FON | Fusarium oxysporum f. sp. niveum |
SIX6 | Secreted in Xylem 6 |
PDA | Potato Dextrose Agar |
PCR | Polymerase Chain Reaction |
BLAST | Basic Local Alignment Search Tool |
FOL | Fusarium oxysporum f. sp. lycopersici |
IGV | Interactive Genomics Viewer |
NCBI | National Center for Biotechnology Information |
HSP | Heat Shock Protein |
ITS | Internal Transcribed Spacer |
COX | Cytochrome c Oxidase |
IGS | Intergenic Spacer |
AVR | Avirulence Gene |
PI | Plant Introduction |
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FON Race | Number of Isolates Tested Using Both Methods | Percentage (%) of Molecular Results Agree with the Bioassay Race Results |
---|---|---|
Race 1 | 29 | 89.65% |
Race 2 | 41 | 80.49% |
Race 3 | 23 | 60.87% |
Assay | Primers | Sequence (5′–3′) | Product Size (bp) | Source |
---|---|---|---|---|
FON specific primer | Fon-1 | CGATTAGCGAAGACATTCACAAGACT | 174 | [19] |
Fon-2 | ACGGTCAAGAAGATGCAGGGTAAAGGT | |||
Race 2 differentiating primer | FONSIX6F | CGCTCTTATCGCATCAATCT | 453 | [3] |
FONSIX6R | GGGTTGACTGAGGTCGTGGT | |||
Race 3 differentiating primer | FNR3F | CGGCTTTCCTCTGTCAGATAGT | 511 | This study |
FNR3R | TAGTGAGGTCCATGCCACGAA |
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Hudson, O.; Waliullah, S.; Fulton, J.C.; Ji, P.; Dufault, N.S.; Keinath, A.; Ali, M.E. Marker Development for Differentiation of Fusarium oxysporum f. sp. Niveum Race 3 from Races 1 and 2. Int. J. Mol. Sci. 2021, 22, 822. https://doi.org/10.3390/ijms22020822
Hudson O, Waliullah S, Fulton JC, Ji P, Dufault NS, Keinath A, Ali ME. Marker Development for Differentiation of Fusarium oxysporum f. sp. Niveum Race 3 from Races 1 and 2. International Journal of Molecular Sciences. 2021; 22(2):822. https://doi.org/10.3390/ijms22020822
Chicago/Turabian StyleHudson, Owen, Sumyya Waliullah, James C. Fulton, Pingsheng Ji, Nicholas S. Dufault, Anthony Keinath, and Md Emran Ali. 2021. "Marker Development for Differentiation of Fusarium oxysporum f. sp. Niveum Race 3 from Races 1 and 2" International Journal of Molecular Sciences 22, no. 2: 822. https://doi.org/10.3390/ijms22020822