A new Rhizobium species that nodulates PhaseoZus vulgaris L. and Leucaena spp. is proposed on the basis of the results of multilocus enzyme electrophoresis, DNA-DNA hybridization, an analysis of ribosomal DNA organization, a sequence analysis of 16s rDNA, and an analysis of phenotypic characteristics. This taxon, Rhizobium tropici sp. nov., was previously named Rhizobium Zeguminosarum biovar phaseoli (type I1 strains) and was recognized by its host range (which includes Leucaena spp.) and nifgene organization. In contrast to R. Zeguminosarum biovar phaseoli, R. tropici strains tolerate high temperatures and high levels of acidity in culture and are symbiotically more stable. We identified two subgroups within R . tropici and describe them in this paper.Members of the genus Rhizobium nodulate the roots of leguminous plants. The rhizobia that infect peas, clovers, and beans (Phaseolus vulgaris L.) are clustered in a single species, Rhizobium leguminosarum (29), which has three biovars (Rhizobium leguminosarum biovar viciae, Rhizobium leguminosarum biovar trifolii, and Rhizobium leguminosarum biovar phaseoli); these biovars contain different symbiotic plasmids that encode distinct nodulation specificities. Nevertheless, heterogeneity in Rhizobium leguminosarum biovar phaseoli has been identified by using such different criteria as protein pattern (50), antibiotic resistance (2), serological type (49), multilocus enzyme electrophoresis behavior (45), DNA-DNA hybridization data (10, 26, 54), plasmid profile (37) , and exopolysaccharide structure (70).We previously distinguished two different types among isolates obtained from bean nodules and found differences in their symbiotic plasmids (36, 38, 39). Type I strains have multiple copies of nitrogenase n i m genes (39, 46), a narrow nodulation host range, and hybridize with the psi (polysaccharide inhibition) gene (3). Type I1 strains have single copies of nifgenes, nodulate Leucaena spp., and do not hybridize with the psi gene (36, 39).Type I1 strains have received attention because their symbiotic plasmids promote an effective and completely differentiated symbiotic process in Agrobacterium tumefaciens recipients (5, 38). They are genetically stable, retaining their symbiotic plasmid after prolonged incubation at 37°C. Some are heat tolerant (31) or acid and aluminum resistant (12, 25, 30, 62). The nodulation genes from one of these strains have been cloned (64). The chemical composition and structure of the extracellular polysaccharides from one type I1 strain differ from the chemical composition and structure of the extracellular polysaccharides from type I isolates (23).Type I1 strains have been less successful in competition for bean nodule occupancy than the type I strains used (41). The former have been reported to occur less frequently in * Corresponding author bean nodules (39). Nodule occupancy by type I1 strains can be enhanced under acid conditions (47, 63).To define the taxonomic position and the genetic relatedness of type I1 strains, we analyzed 64 typ...

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Screening of Rhizobium leguminosarum bv. phaseoli strains showed some that were able to nodulate common beans (Phaseolus vulgaris L.) at high temperatures (35 and 38°C/8h/day). The nodulation ability was not related to the capability to grow or produce melanin-like pigment in culture media at high temperatures. However, nodules formed at high temperatures were ineffective and plants did not accumulate N in shoots. Two thermal shocks of 40°C/8 h/day at flowering time drastically decreased nitrogenase activity and nodule relative efficiency of plants otherwise grown at 28°C. Recovery of nitrogenase activity began only after seven days, when new nodules formed; total incorporation of N in tops did not recover for 2 weeks. Non-inoculated beans receiving mineral N were not affected by the thermal shock, and when growing continuously at 35 or 38°C had total N accumulated in shoots reduced by only 18%.

SUMMARYAn extensive search for active (acetylene reducing) nitrogen-fixing root nodules was made amongst Brazilian forest legumes. Seven new nodulated species, including two new nodulating genera, were found in the Caesalpinioideae, 18 new species in the Mimosoideae and 30 new species, including four new genera, in the Papilionoideae. All have potential importance in reafforestation. Fifty-eight new records of non-nodulating species are included. The morphology of all nodules was studied and is discussed in relation to taxonomic position. Preliminary data on nodule structure are given.

SUMMARYAeschynomene fluminensis Veil., originally obtained from flooded areas of the Pantanal Matogrossense region of Brazil, was grown under stem-flooded or non-flooded conditions for 70 d after inoculation with isolates of photosynthetic stem nodule rhizobia obtained from native A. fluminensis. Stem nodules formed only on submerged stems of flooded plants (mean of 25 per plant), and did not form on aerial parts, although they were capable of growing and fixing N^ after drainage of the stems. Root nodules formed on both non-flooded and flooded plants but were usually decreased in number by flooding (from means of 124 to 51 per plant, respectively). Flooding (and stem-nodulation) resulted in an increase in shoot (and a decrease in root) dry weight, regardless of rhizobial isolate.Stem nodules were attached by a wide collar of aerenchymatous tissue at the base of the nodule. There were large air spaces in the stem where nodules were subtended and these were continuous with nodule aerenchyma/outer corte.x. In addition, aerenchyma and spongy tissue at the base of the nodule connected both flooded and non-flooded root nodules to large intercellular spaces in the root cortex. The stem and root nodules were ovoid in shape, and essentially aeschynomenoid in type, i.e. the central infected tissue was without uninfected, interstitial cells. Root nodules had a similar structure to stem nodules (although stem nodules were generally larger), and flooded root nodules were approximately twice the size of non-flooded nodules. The infected tissue of root and stem nodules consisted of spherical, bacteroid-containing cells containing one or two rod-shaped bacteroids per peribacteroid unit and prominent organelles. Infection threads were observed in root but not in stem nodules.The cortex of stem and root nodules had an apparent oxygen diffusion barrier, consisting of concentric layers of small cells with interlocking cell walls and few intercellular spaces. Cell layers external to these consisted of larger cells and intercellular spaces, with some spaces being occluded with an electron-dense material that contained a glycoprotein recognized by the monoclonal antibodies MAC236 and MAC265. The amount of glycoprotein occlusions did not appear to differ between nodule types or treatments, although stem nodules contained intracellular glycoprotein vesicles adjacent to cell walls. The exterior of the nodules consisted of an epidermis of thin flattened cells with occasional lenticels. Amyloplasts were common in lower stem and hypocotyl nodules, but fewer in flooded or non-flooded root nodules. Upper stem nodules (i.e. those within 6 cm of the water surface) differed from more profoundly submerged stem nodules by having chloroplasts throughout the cortex. Root nodules did not contain chloroplasts, and undifferentiated plastids were found mainly in lower stem nodules.

RESUMOOs objetivos do presente estudo foram avaliar a influência de Acacia auriculiformis (acácia), Mimosa caesalpiniifolia (sabiá) e Corymbia citriodora (eucalipto), comparadas às coberturas de capoeira e pastagem, sobre os atributos químicos e microbianos do solo. As espécies apresentaram sobrevivência no campo acima de 70 % e bom desenvolvimento em altura e diâmetro (DAP). Entre as leguminosas, o sabiá apresentou a maior taxa de sobrevivência, enquanto a acácia mostrou melhor desenvolvimento, especialmente entre os 27 e 44 meses. As espécies florestais, na fase inicial de desenvolvimento, revelaram-se promissoras em melhorar a fertilidade do solo, apesar da redução dos teores de matéria orgânica. A pastagem mostrou potencial de manter o teor de MO e melhorar a fertilidade do solo. O C e N microbiano foram mais discriminantes que o C orgânico do solo às mudanças na cobertura vegetal. Os índices microbianos (qCO 2 -quociente metabólico, CBM/C -relação C da biomassa microbiana/C orgânico, NBM/N -relação N da biomassa microbiana/N total e relação C/N microbiana) foram mais discriminantes que os atributos químicos (C orgânico e N total) e microbianos (CBM -carbono da biomassa microbiana, NBM -N da biomassa microbiana e RArespiração acumulada) em aferir a dissimilaridade entre as coberturas vegetais.