In this study, we identified disease resistance gene homologs in Brassica oleracea and assessed their expression in lines resistant and susceptible to Xanthomonas campestris pv. campestris (Xcc). Two DNA fragments of approximately 2.5 kb (BI-16/RPS2 and Lc201/RPS2) were amplified by PCR from two Brassica lines using primers based on an RPS2 homologous sequence previously described in the Brassica oleracea ecotype B117. The sequences of these fragments shared high similarity (95-98%) with RPS2 homologs from various Brassica species. The digestion of these fragments with restriction enzymes revealed polymorphisms at the Xba I restriction sites. The length polymorphisms were used as a co-dominant marker in an F2 population developed to segregate for resistance to Xcc, the causal agent of black rot. Linkage analysis showed no significant association between the marker and quantitative trait loci for black rot. RT-PCR with specific primers yielded an expected 453 bp fragment that corresponded to the RPS2 homologs in both resistant and susceptible lines inoculated with the pathogen, as well as in non-inoculated control plants. These results suggest that these homologs are constitutively expressed in B. oleracea.
The effectiveness of Bradyrhizobium strains with characteristics Hup+ (SR and USDA-110), Hup- (29W) and Hup hr (SEMIA-587), was evaluated in cowpea (Vigna unguiculata L.), cultivars IPA-202, BR-3 and VITA-4. Results showed that nodulation in VITA-4 was superior, with a positive interaction when inoculated with strains SEMIA-587 and USDA-110. However, when comparing nodule efficiency, the combination IPA-202 x SEMIA-587 presented the highest nitrogenase activity, with a relative efficiency around 1.0. Nitrogenase activities were similar in plants inoculated with the strains SR and SEMIA-587, and higher, with strains USDA-110 and 29W. These results indicate that strains with Hup+ and Hup hr were able to achieve higher enzymatic activities. Leghemoglobin (Lb) content detected in strains SR and USDA-110 was positively correlated to nitrogenase activity. However, correlations between Lb and nitrogenase activity were highly variable in nodules containing strain 29W-Hup-, indicating that imbalances in the nitrogenase system in the absence of hydrogenase may affect the flow of Lb to the bacteroids. Significant differences on the plant N content were found among the strains studied, but not among cultivars. Plant inoculated with Hup+ strains were more efficient to accumulate N, indicating that N-fixing systems retaining H2 may accumulate additional N in the host.
A eficiência das estirpes de Bradyrhizobium com características Hup+ (SR e USDA-110), Hup- (29W) e Hup hr (SEMIA-587) foi avaliada em caupi (Vigna unguiculata L.), cultivares IPA-202, BR-3 e VITA-4. Os resultados mostraram que VITA-4, em relação à nodulação, revelou-se superior às demais, e apresentou interação efetiva com as estirpes SEMIA-587 e USDA-110. Entretanto, quanto à eficiência nodular, a combinação IPA-202 x SEMIA-587 alcançou maior atividade da nitrogenase (ARA) com eficiência relativa próxima a 1,0. A ARA detectada nas estirpes SR e SEMIA-587 foi similar, porém, superior às estirpes USDA-110 e 29W, evidenciando que as estirpes Hup+ e Hup hr alcançaram maiores atividades enzimáticas. Os teores de leghemoglobina (Lb) detectados nas estirpes SR e USDA-110 foram positivamente relacionados com as respectivas ARA, contudo, a relação entre teor de Lb e ARA obtido para 29W-Hup- foi variável, sugerindo que, na ausência da hidrogenase, o sistema da nitrogenase fica afetado podendo influir no fluxo de Lb ao bacteróide. A avaliação do teor de N mostrou que não houve diferença entre cultivares, entretanto, foi detectada diferença significativa entre as estirpes. As estirpes Hup+ obtiveram maiores acúmulos de N, evidenciando que os sistemas simbióticos que menos liberam H2, acumulam mais N no hospedeiro.