Chromosome doubling was induced in vitro in a diploid hybrid of Rosa rugosa Thunb. using oryzalin as the spindle inhibitor. Nodal sections, 2 mm long, were exposed to 2.5 or 5 microM oryzalin and 10 mm nodal sections were exposed to 5 microM oryzalin for 0 (controls), 6, 12, 24 and 48 h. The ploidy of the emergent shoots was determined by flow cytometry. The frequency of tetraploid and mixoploid leaves that developed from 2 mm nodal sections exposed to 5 microM oryzalin peaked at 12 h exposure, when 35% of the leaves were tetraploid, but fell after longer exposures. Fewer tetraploid and mixoploid leaves were found when 2 mm nodes were exposed to 2.5 microM oryzalin for 6 and 12 h, indicating that it took longer for a spindle inhibiting concentration of oryzalin to build up in the meristem. However, the frequencies of tetraploid and mixoploid leaves continued to rise after 12 h and were highest at 48 h, when 44% were tetraploid. In treatments with 5 microM oryzalin, the frequencies of tetraploid and mixoploid leaves were lower, at equivalent exposure times, in 10 mm nodes than 2 mm nodes. This suggests that oryzalin diffused to the meristem mainly via the cut surfaces and that access via the epidermis and cuticle was impeded.

The interactions of four pathotypes of Diplocarpon rosae with 34 species and hybrids of Rosa were compared in an ongoing search for criteria of potential relevance to rose breeding. There were greater similarities in the resistance-susceptibility interactions of these roses to the pathotypes DA1, CW1 and ZM1, than to a fourth pathotype, DA2. Among the species roses, only those of the section Cinnamomeae were susceptible to DA2 and this susceptibility was transmitted in several cases to hybrid progeny. CW1 had the greatest pathogenicity of the four pathotypes. In a cross between R. rugosa cv. Scabrosa, which was resistant and R. rugosa f. alba, which was susceptible to all pathotypes, the progeny were tested for resistance to pathotypes DA1, CW1 and ZM1. Each of the 20 progeny was susceptible to all three pathotypes. This shows that, unlike the well characterized Rdr1 gene for resistance to D. rosae, the resistance of R. rugosa cv. Scabrosa is not determined by a dominant major gene. The diploid hybrid, R. rugosa cv. Martin Frobisher · cv. Mistress Quickly, was resistant to all four pathotypes, but an induced tetraploid of this hybrid was susceptible to all pathotypes. The relevance of these findings to the breeding of roses for resistance to black spot disease is discussed.