RESUMO
ABSTRACT Until recently, tomato race 1 (T1) of Xanthomonas campestris pv. vesicatoria was the only race causing bacterial spot of tomato in Florida. In 1991, tomato race 3 (T3) was first identified in 3 of 13 tomato production fields surveyed. By 1994, T3 was observed in 21 of 28 fields and was the only race identified in 14 fields. In field studies, tomato genotypes with resistance to either T1 or T3 or susceptibility to both were co-inoculated with strains of both races. Lesions on 10 plants in each of three replications for each genotype were sampled three times during the experiment; bacterial isolations were made from each lesion, and tomato race identifications were made for each strain. At the third sampling date, T3 was isolated from 97% of the lesions on the susceptible genotype Walter and the T1-resistant genotype Hawaii 7998, while T3 was isolated from 23% of the lesions and T1 from the remaining 77% on the T3-resistant genotypes PI 128216 and PI 126932. In surface population studies done in growth rooms, suspensions of T1 and T3 were applied alone and in combination to the leaf surfaces of susceptible and resistant genotypes. T1 populations were reduced more than 10-fold when applied in combination with T3, compared with populations that developed when T1 was applied alone. T3 populations were not affected when applied in combination with a T1 strain. In greenhouse studies with the T3-resistant genotype Hawaii 7981, disease was significantly reduced in plants inoculated with T3 in combination with T1, compared with plants inoculated with T1 alone. These results clearly demonstrate the competitive nature of T3 in the presence of T1 and help explain the emergence of T3 as a prevalent race in Florida.
RESUMO
In vitro and in planta sensitivity of an indirect enzyme-linked immunoassay technique, using a monoclonal antibody specific for the lipopolysaccharide (LPS) of Xanthomonas campestris pv. vesicatoria, was increased 10-fold by using a new extraction buffer (gl of: KH2PO4, 2; NaHPO4, 11.5; EDTA disodium, 0.14; thimerosal, 0.02; and lysozyme, 0.2). The procedure improved sensitivity without increasing background levels. In vitro, the limit of detection was between 1 x 10(7) and 1 x 10(8) cells ml-1 with the conventional extraction buffer phosphate-buffered saline (PBS) and less than 1 x 10(6) cells ml-1 when lysozyme extraction buffer was substituted for PBS. In comparing 22 X. c. vesicatoria strains, absorbance readings were increased close to three-fold with the lysozyme extraction buffer as opposed to PBS. When leaf tissue extract was spiked with the bacterium, the limit of detection was 1 x 10(7) cfu ml-1 and 1 x 10(8) cfu ml-1 with the lysozyme solution and PBS, respectively, as the extraction buffers. When using the lysozyme extraction buffer in combination with a commercial amplification system, the limit of detection was decreased to less than 1 x 10(5) cfu ml-1 in leaf tissue. The addition of the lysozyme and EDTA to the phosphate buffer resulted in release of a significant quantity of LPS and concomitant dramatic increase in sensitivity. The new procedure, termed lysozyme ELISA (L-ELISA), should increase sensitivity of ELISA reactions where LPS is the reacting epitope.