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CROP BREEDING & GENETICS

AFLP and SCAR Markers Linked to Tomato Spotted Wilt Virus Resistance in Tobacco

Dep. of Crop Science, North Carolina State Univ., Box 7620, Raleigh, NC 27695-7620

^* Corresponding author (jennifer_nicholson{at}ncsu.edu).

Tomato spotted wilt virus (TSWV) is a serious disease in tobacco (Nicotiana tabacum L.). The breeding line ‘Polalta’ contains a single dominant gene conferring resistance to TSWV that was introgressed from N. alata Link & Otto. The resistance is tightly associated with an abnormal plant type, however, and traditional backcrossing has been ineffective in producing normal plants with TSWV resistance. A potential strategy to overcome this problem is to use molecular markers to select against alien chromatin in backcross progeny. Amplified fragment length polymorphism (AFLP) technology and bulked segregant analysis were applied to identify markers linked to the resistance gene. The DNA bulks from susceptible and resistant doubled haploid lines derived from a cross between susceptible cultivar K326 and Polalta were analyzed to identify linked markers. An F₂ population and the doubled haploids were used to construct a 2.5-cM map of this locus containing 17 coupling phase and seven repulsion phase markers. By selecting for resistant plants with an improved plant type and marker profile in a BC₃ population, four plants were identified that were missing 1 to 14 of the AFLP markers associated with the alien chromatin, demonstrating that rare recombination events can be identified. Four AFLP fragments were successfully converted to sequence characterized amplified region markers suitable for large-scale screening. This approach may facilitate development of TSWV-resistant tobacco cultivars.

Abbreviations: AFLP, amplified fragment length polymorphism • PCR, polymerase chain reaction • SCAR, sequence characterized amplified region • TSWV, Tomato spotted wilt virus