Walnut blackline disease, caused by the walnut strain of the cherry leafroll virus, causes fatal necrosis of the graft union between susceptible, infected scions of Persian walnut (Juglans regia L.) and hypersensitive, resistant rootstocks. A backcross breeding program to transfer hypersensitivity to cherry leafroll virus from the Norther California black walnut (Juglans hindsii (Jeps.)), into Persian walnut was begun in 1987. Hypersensitivity to the virus is inherited as a single, dominant gene. The current procedures for identifying hypersensitive backcross progeny are slow and labor intensive. Bulks of DNA from backcrosses that were either hypersensitive or susceptible to cherry leafroll virus were compared using randomly amplified polymorphic DNA. One random decamer, sequence 5'-CTCCTGCCAA-3' (OP-K15), produces a polymorphic fragment of about 720 bp that has about 7% recombination with hypersensitivity to cherry leafroll virus in our backcross populations. The polymorphic fragment was cloned and converted into a restriction fragment length polymorphism to demonstrate that it is a distinct, low-copy sequence.