Sugar cane (Saccharum spp.) variety SP 70-1143 was inoculated with Gluconacetobacter diazotrophicus strain PAL5 (ATCC 49037) in two experiments. In experiment 1 the bacteria were inoculated into a modified, low sucrose MS medium within which micropropagated plantlets were rooted. After 10 d there was extensive anatomical evidence of endophytic colonization by G. diazotrophicus, particularly in lower stems, where high numbers of bacteria were visible within some of the xylem vessels. The identity of the bacteria was confirmed by immunogold labelling with an antibody raised against G. diazotrophicus. On the lower stems there were breaks caused by the separation of the plantlets into individuals, and at these 'wounds' bacteria were seen colonizing the xylem and intercellular spaces. Bacteria were also occasionally seen entering leaves via damaged stomata, and subsequently colonizing sub-stomatal cavities and intercellular spaces. A localized host defence response in the form of fibrillar material surrounding the bacteria was associated with both the stem and leaf invasion. In experiment 2, stems of 5-week-old greenhouse-grown plants were inoculated by injection with a suspension of G. diazotrophicus containing 10(8) bacteria ml(-1). No hypersensitive response (HR) was observed, and no symptoms were visible on the leaves and stems for the duration of the experiment (7 d). Close to the point of inoculation, G. diazotrophicus cells were observed within the protoxylem and the xylem parenchyma, where they were surrounded by fibrillar material that stained light-green with toluidine blue. In leaf samples taken up to 4 cm from the inoculation points, G. diazotrophicus cells were mainly found within the metaxylem, where they were surrounded by a light green-staining material. The bacteria were growing in relatively low numbers adjacent to the xylem cell walls, and they were separated from the host-derived material by electron-transparent 'haloes' that contained material that reacted with the G. diazotrophicus antibody.

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-The aim of this work was to investigate the effect of water stress on N 2 fixation and nodule structure of two common bean (Phaseolus vulgaris L.) cultivars Carioca and EMGOPA-201. Plants were harvested after five and eight days of water stress. Carioca had lower nodule dry weight on both water stress periods; shoot dry weight was lower at five days water stress and did not differ from control after eight days stress. Both cultivars had lower nitrogenase activity than control after five and eight days water stress. For both cultivars, after eight days stress bacteroid membranes were damaged. Carioca presented more pronounced damage to infected tissue, with host cell vacuolation and loss of the peribacteroid membrane at five days after stress; at eight days after stress, there was degradation of cytoplasm host cells and senescence of bacteroids, with their release into intercellular spaces. Intensity of immunogold-labeling of intercellular cortical glycoprotein with the monoclonal antibodies MAC 236/ 265 was different for both cultivars.Index terms: Phaseolus vulgaris, nitrogenase, leaf water potential, nodulation. Efeito do estresse hídrico na fixação biológica de nitrogênio e estrutura de nódulos de feijãoResumo -O objetivo deste trabalho foi investigar o efeito do estresse hídrico na fixação biológica do N 2 e na estrutura dos nódulos de feijão (Phaseolus vulgaris L.), nas cultivares Carioca e EMGOPA-201. As plantas foram colhidas após cinco e oito dias de estresse hídrico. A cultivar Carioca apresentou menor peso seco de nódulos que o controle (plantas irrigadas). O peso da parte aérea seca da cultivar Carioca foi menor aos cinco dias de estresse hídrico e não diferiu do controle, após oito dias de estresse. As duas cultivares tiveram menor atividade da nitrogenase após cinco e oito dias de estresse hídrico e apresentaram danos na membrana peribacteróide após oito dias de estresse. A cultivar Carioca foi mais afetada do que a EMGOPA-201, com vacuolação das células e perda da membrana peribacteróide após cinco dias de estresse; aos oito dias, houve degradação das células do citoplasma, senescência e liberação dos bacteróides nos espaços intercelulares. A intensidade de glicoproteínas nos espaços intercelulares do córtex, marcada com os anticorpos MAC 236/265, foi diferente nas duas cultivares de feijão.Termos para indexação: Phaseolus vulgaris, nitrogenase, potencial de água na folha, nodulação.