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The objective of this study was to evaluate the effects of strains of Azospirillum brasilense, Pseudomonas fluorescens and Rhizobium tropici on the shoot dry weight (SDW) and root dry weight (RDW) yield, N uptake and nutritive value of 'Mavuno' grass inoculated with plant growth-promoting bacteria. We evaluated the effects of inoculation with the Ab-V5 and Ab-V6 strains of Azospirillum brasilense and Pseudomonas fluorescens or co-inoculation with Rhizobium tropici and Ab-V6, with and without nitrogen (N) application, as well as re-inoculations. The growth promoting bacteria + N promoted increases in SDW and RDW yield, tillers dry weight, relative chlorophyll index and N uptake. There were no effects of re-inoculation by Azospirillum brasilense, Pseudomonas fluorescens and Rhizobium tropici on nutrition, nutritive value and SDW and RDW yield, demonstrating that this technique still needs further studies with 'Mavuno' grass in the form and the correct period to be performed. [ABSTRACT FROM AUTHOR]

Biological N2 fixation can fulfil the N demand of legumes but may cost as much as 14% of current photosynthate. This photosynthate (C) sink strength would result in loss of productivity if rates of photosynthesis did not increase to compensate for the costs. We measured rates of leaf photosynthesis, concentrations of N, ureides and protein in leaves of two soybean cultivars ( Glycine max [L.] Merrill) differing in potential shoot biomass production, either associated with Bradyrhizobium japonicum strains, or amended with nitrate. Our results show that the C costs of biological N2 fixation can be compensated by increased photosynthesis. Nodulated plants shifted N metabolism towards ureide accumulation at the start of the reproductive stage, at which time leaf N concentration of nodulated plants was greater than that of N-fertilized plants. The C sink strength of N2 fixation increased photosynthetic N use efficiency at the beginning of plant development. At later stages, although average protein concentrations were similar between the groups of plants, maximum leaf protein of nodulated plants occurred a few days later than in N-fertilized plants. The chlorophyll content of nodulated plants remained high until the pod-filling stage, whereas the chlorophyll content of N-fertilized plants started to decrease as early as the flowering stage. These results suggest that, due to higher C sink strength and efficient N2 fixation, nodulated plants achieve higher rates of photosynthesis and have delayed leaf senescence. [ABSTRACT FROM AUTHOR]

Drosera villosa var. villosa A. St.-Hil is a carnivorous plant that grows in Brazilian flooded soils very poor on nutrients, including low levels of N. Under these conditions, the plant shows vigorous growth, low root number, low number of captured prey (less than 50%) and a great assemblage of bacteria associated with the roots and leaves that grow in N-free medium. These preliminary results have led us to investigate the number of colony forming units (log CFU) in the roots (rhizosphere and endorhizosphere) and leaves (phyllosphere and endophyllosphere) of D. villosa var. villosa by the tenfold serial dilution technique in two N-free culture media. The results showed that the phyllosphere had 6.65 log CFU g dry leaf−1 and the rhizosphere 6.47 log CFU g dry soil−1, with the lowest value detected in the endophyllosphere (4.39 log CFU g dry leaf−1). Sixty-three different bacteria morphotypes were isolated from the surface and interior of the roots and leaves and the amplification of the DNA with specific primers detected the nifH gene in 34 of those strains. The DNAs of the 34 strains were compared by the BOX-PCR technique and a great diversity was observed, with the bacteria clustering at a final level of similarity of only 12%. The strains were also submitted to the partial sequencing of the 16S rRNA gene and several genera of N2-fixing bacteria were detected, including Bacillus, Burkholderia, Methylobacterium, Paenibacillus, Pseudomonas and Sphingomonas. [ABSTRACT FROM AUTHOR]

M. Hungria, S. Astolfi-Filho, L.M.O. Chueire, M.F. Nicolás, E.B.P. Santos, M.R. Bulbol, A. Souza-Filho, E. Nogueira Assunção, M.G. Germano and A.T.R. Vasconcelos. 2005. To isolate and to characterize the diversity of Chromobacterium violaceum from the Brazilian Amazon region. Twenty-two isolates were obtained from the waters and banks of the river Negro, in the Brazilian Amazon. All isolates were able to grow in vitro at 44°C and pH 4·0, but were adversely affected by temperatures below 15°C, and unable to survive at 4°C, properties that may be related to the adaptation to the ecosystem. The isolates were joined at a final level of similarity of only 13% in the rep-PCR analysis. The analysis of 16S rRNA genes resulted in three main groups clustered at a final level of similarity of 97% and only three isolates were clustered with the type strain. Similar data were obtained for the 23S rRNA gene. A high level of genetic diversity was verified with indications that the Brazilian isolates would fit into at least two new clusters besides C. violaceum species. The results show remarkable bacterial adaptability and genetic diversity of C. violaceum in the Amazon region. [ABSTRACT FROM AUTHOR]

In order to identify soybean cultivars with higher biological N2 fixation capacities, North American and Brazilian soybean [G lycine max (L.) Merrill] cultivars, belonging to maturity groups VI–VIII, were evaluated for nodulation parameters and N2 fixation rates. The symbiotic performance of 152 cultivars was evaluated in pots containing 4 kg soil with an established population of the three Bradyrhizobium elkanii strains [29w (SEMIA 5019):SEMIA 566 : SEMIA 587, 22%:36%:34%] which are established in most Brazilian soils cultivated with soybean. Differences were verified among cultivars, with some accumulating up to twice as much nodule dry weight and N in tissues as others. The variability among cultivars was also confirmed when six of them were used in a field experiment, resulting in differences in nodulation, yield and total N accumulated in grains. The analysis of nodule occupancy in 12 cultivars grown either under sterile conditions and receiving a double inoculum and N-free nutrient solution, or in pots containing soil with an established population of bradyrhizobia, showed the preference of cultivars for specific strains. [ABSTRACT FROM AUTHOR]

In a previous study soybean Bradyrhizobium strains, used in Brazilian studies and inoculants over the last 30 years, and strains adapted to the Brazilian Cerrados, a region frequently submitted to environmental and nutritional stresses, were analyzed for 32 morphological and physiological parameters in vivo and in vitro. A cluster analysis allowed the subdivision of these strains into species Bradyrhizobium japonicum, Bradyrhizobium elkanii and a mixed genotype. In this study, the bacteria were analyzed for nodulation, N2 fixation capacity, nodule occupancy and the ability to increase yield. The goal was to find a relationship between the strain groups and the symbiotic performance. Two strains of Brazilian B. japonicum showed higher rates of N2 fixation and nodule efficiency (mg of N mg–1 of nodules) under axenic conditions. These strains also showed greater yield increases in field experiments when compared to B. elkanii strains. However, no differences were detected between B. japonicum and B. elkanii strains when comparing nodule occupancy capacity. The adapted strains belonging to the serogroup B. elkanii SEMIA 566, most clustered in a mixed genotype, were more competitive than the parental strain, and some showed a higher capacity of N2 fixation. Some of the adapted strains, such as S-370 and S-372, have shown similar N2 fixation rates and nodulation competitiveness to two Brazilian strains of B. japonicum. This similarity demonstrates the possibility of enhancing N2 fixing ability, after local adaptation, even within B. elkanii species. Differences in the DNA profiles were also detected between the parental SEMIA 566 and the adapted strains by analyses with the ERIC and REP-PCR techniques. Consequently, genetic, morphological and physiological changes can be a result of adaptation of rhizobia to the soil. This variability can be used to select strains capable of increasing the contribution of N2 fixation to soybean nutrition. [ABSTRACT FROM AUTHOR]

Several years of research have shown that there is a high genetic and physiological variability among Bradyrhizobium japonicum strains, culminating in a subdivision into two bacterial genotypes, and the description of the new species B. elkanii. In Brazil, large-scale soybean inoculation started in 1960 and today 15 million doses of inoculants are sold per year for an estimated area of 12 million ha. Efforts have been made to find strains able to fix high amounts of N2 under Brazilian soil conditions, but few laboratories cover basic studies on N2 fixation, such as strain classification into the two Bradyrhizobium species. In this study several characteristics of 40 soybean Bradyrhizobium strains, including 4 reference strains of B. japonicum (genotype I) species, 3 of B. elkanii (genotype II) and 1 of a mixed genotype were evaluated. The parameters analysed in vitro were: colony morphology, serological grouping, intrinsic resistance to antibiotics, synthesis of indole acetic acid, expression of hydrogenase activity and growth in a medium enriched with asparagine. In vivo, analyses performed included the nodulation of Rj 4 soybean cultivar Hill and the detection of symptoms caused by rhizobitoxine. These evaluations allowed a phenotypic grouping which positioned most of the strains utilized in Brazilian inoculants and studies, as well as some new strains isolated from the Cerrado region, within the species B. elkanii. However, environmental stresses and adaptation of Bradyrhizobium strains to the soil caused a large physiological and genetic variability in some isolates from the Cerrado soils in relation to the putative parental strain introduced 15 years ago, placing these isolates in an intermediate position between the two Bradyrhizobium species. [ABSTRACT FROM AUTHOR]

Differences between isogenic uptake hydrogenase (HUP) mutants of Bradyrhizobium japonicum in terms of nodule efficiency, N fixation and N incorporation into various plant parts were studied in a monoxenic greenhouse experiment in order to confirm previous results with soybeans and beans inoculated with various HUP and HUP strains. The HUP+ revertant PJ17-1 of a HUP mutant (PJ17) of strain USDA DES 122 showed a completely restored relative efficiency (100% versus 78±2% for the HUP mutant), higher nodule efficiency (N fixed per g nodules), higher ureide-N transport rates, higher N contents in pods and higher N harvest indices. All these observations confirm previous experiments with HUP and HUP strains. [ABSTRACT FROM AUTHOR]

In an experiment performed under greenhouse conditions, four cultivars of Phaseolus vulgaris L. (Venezuela-350; Aroana; Moruna; Carioca) were inoculated with three Rhizobium leguminosarum biovar phaseoli strains (C-05; C-40 = CIAT 255; C-89 = CIAT 55) and were fertilized with an N-free mineral nutrient solution. The plants were harvested 25, 40, and 55 days after emergence and the following paramenters were evaluated: Nitrogenase activity of nodulated roots, H evolution by the nodules; relative efficiency of nitrogenase; respiration rates of nodulated roots and detached nodules; dry weight and total N of stems, leaves, pods, roots, and nodules. Generally the bean cultivar, Rhizobium strain, had an effect and there was an interaction effect with both symbiotic partners, on all parameters. On average, nodules represented 23% of total root respiration but the best symbiotic combinations showed lower ratios of C respired to N fixed. The maximum N-assimilation rate (between 40 and 55 days after emergence) of 11.93 mg N plant day occurred with the symbiotic combination of Carioca × C-05, while the poorest rate of 0.55 mg N plant day was recorded with Venezuela-350 × C-89. The best symbiotic combinations always showed the highest relative nitrogenase efficiency, but the differences in N-fixation rates cannot be explained solely in terms of conservation of energy by recycling of H. This requires further investigation. [ABSTRACT FROM AUTHOR]

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