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CROP BREEDING, GENETICS & CYTOLOGY

Efforts to Initiate Construction of a Disease Resistance Package on a Designer Chromosome in Tobacco

Department of Crop Science, North Carolina State University, Raleigh, NC 27695-7620

* Corresponding author (ewernsma{at}cropserv1.cropsci.ncsu.edu)

Gene cloning and transformation can be used to circumvent linkage drag effects that can plague conventional interspecific gene transfers. These techniques can also be used to create desirable genetic linkages. Use of Nicotiana glutinosa L. N-gene mediated TMV (tobacco mosaic virus) resistance in flue-cured tobacco, N. tabacum L., has been limited due to linkage drag effects. Transformation was used to introduce the cloned N-gene into NC152, a chromosome addition line possessing a chromosome pair from N. africana. This chromosome has been proposed to be used as a "designer chromosome" into which numerous transgenes could be inserted to form a desirable linkage package. The system could be used to shuttle a large number of transgenes from genotype to genotype. One hundred thirty-six primary transformants possessing the N transgene were produced and hybridized with TMV-susceptible ‘Petite Havana.’ These may serve as valuable TMV-resistant breeding materials. For each independent transformant, BC₁F₁ families which segregated for TMV resistance and the addition chromosome were generated. Data from cosegregation, transmission, and molecular analyses were used to conclude that one transformant possessed an insertion of the N-gene in the addition chromosome. By inserting N in the chromosome, we initiated construction of a disease resistance package by linking the TMV resistance gene with a potyvirus resistance gene(s) native to the chromosome. Occasional loss of the transgene, however, may be evidence of previously undetected interchromosomal recombination, and may have implications for use of this system in cultivar development.

Abbreviations: bp, base pairs • Mtx, methotrexate • PCR, polymerase chain reaction • PVY, potato virus Y • TEV, tobacco etch virus • TMV, tobacco mosaic virus

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