Comparison of colony morphology, salt tolerance, and effectiveness in Rhizobium japonicum

1977 ◽  
Vol 23 (9) ◽  
pp. 1118-1122 ◽  
Author(s):  
Robert G. Upchurch ◽  
Gerald H. Elkan

Four strains of Rhizobium japonicum, two of which produce slimy and non-slimy colony types and two others which produce large and small colony types, were isolated and cloned. All were infective and nodulated Lee soybean host plants. Each colony type was characterized as to its salt sensitivity to Na+ and K+ ions, relative level of symbiotic nitrogen fixation, and relative level of free-living nitrogen fixation. Growth studies performed in the presence of salts demonstrated that the non-slimy or small colony types were sensitive to salt with significantly depressed growth rates and cell yields. Growth rates and cell yields of slimy, large, colony types were relatively unaffected by salt. Both symbiotic and free-living (non-associative) nitrogen fixation analyses (by acetylene reduction) revealed that the non-slimy, small colonies were significantly more effective than slimy, large colonies.

1988 ◽  
Vol 34 (8) ◽  
pp. 1017-1022 ◽  
Author(s):  
Jeffrey S. La Favre ◽  
Adrienne K. La Favre ◽  
Allan R. J. Eaglesham

Bradyrhizobium japonicum strain USDA 76, a rhizobitoxine producer, was found to contain two colony types, designated "small" and "large" based on colony size on agar medium. Only the small type produced detectable chlorosis-inducing toxin in culture, whereas both colony types induced chlorosis as a result of synthesis of toxin in nodules. Electron microscopy revealed that a large colony derivative, grown in broth culture, was encapsulated, whereas a small colony derivative was not, suggesting a negative functional relationship between toxin synthesis and presence of capsule. The large type also had a ruthenium red reactive extracellular layer when cultured in the soybean rhizosphere. This differential production of toxin by the colony derivatives in culture, and presumably in the rhizosphere, prompted the investigation of a proposed role of rhizobitoxine in the infection of roots of nodulation-refractory (rj1rj1) soybean; the small colony type formed fewer nodules on the (rj1rj1) isoline, indicating no major function for rhizobitoxine in the infection of (rj1rj1) soybean.


2003 ◽  
Vol 185 (24) ◽  
pp. 7266-7272 ◽  
Author(s):  
Wen-Ming Chen ◽  
Lionel Moulin ◽  
Cyril Bontemps ◽  
Peter Vandamme ◽  
Gilles Béna ◽  
...  

ABSTRACT Following the initial discovery of two legume-nodulating Burkholderia strains (L. Moulin, A. Munive, B. Dreyfus, and C. Boivin-Masson, Nature 411:948-950, 2001), we identified as nitrogen-fixing legume symbionts at least 50 different strains of Burkholderia caribensis and Ralstonia taiwanensis, all belonging to the β-subclass of proteobacteria, thus extending the phylogenetic diversity of the rhizobia. R. taiwanensis was found to represent 93% of the Mimosa isolates in Taiwan, indicating thatβ -proteobacteria can be the specific symbionts of a legume. The nod genes of rhizobial β-proteobacteria (β-rhizobia) are very similar to those of rhizobia from theα -subclass (α-rhizobia), strongly supporting the hypothesis of the unique origin of common nod genes. Theβ -rhizobial nod genes are located on a 0.5-Mb plasmid, together with the nifH gene, in R. taiwanensis and Burkholderia phymatum. Phylogenetic analysis of available nodA gene sequences clustered β-rhizobial sequences in two nodA lineages intertwined with α-rhizobial sequences. On the other hand, theβ -rhizobia were grouped with free-living nitrogen-fixingβ -proteobacteria on the basis of the nifH phylogenetic tree. These findings suggest that β-rhizobia evolved from diazotrophs through multiple lateral nod gene transfers.


1965 ◽  
Vol 11 (1) ◽  
pp. 29-38 ◽  
Author(s):  
P-C. Chang ◽  
R. Knowles

The occurrence of free-living nitrogen fixers, the potential for nitrogen fixation, and the correlation between the nitrogen-fixing capacities of the soils and bacterial counts were studied using representative Quebec soils.Clostridium occurred more frequently than did Azotobacter. Studies with N15showed that nitrogen fixation was more frequent under anaerobic than under aerobic conditions in all the soil types studied in their unamended state. The addition of glucose stimulated nitrogen fixation. During anaerobic incubation, nitrogen fixation was found to be correlated significantly with the increase in numbers of both total aerobes and Clostridia. The results suggested that facultatively anaerobic nitrogen fixers, and aerobic nitrogen fixers other than Azotobacter, were present.


Author(s):  
G. C. Machray ◽  
W. D. P. Stewart

SynopsisA wide variety of plant-microbe nitrogen-fixing symbioses which include cyanobacteria as the nitrogenfixing partner exist. While some information has been gathered on the biochemical changes in the cyanobacterium upon entering into symbiosis, very little is known about the accompanying changes at the genetic level. Much of our present knowledge of the organisation and control of expression of nitrogenfixation (nif) genes is derived from studies of the free-living diazotroph Klebsiella pneumoniae. This organism thus provides a model system and source of experimental material for the genetic analysis of symbiotic nitrogen fixation. We describe the use of cloned K. pneumoniae genes for nitrogen fixation and its regulation in the genetic analysis' of nitrogen fixation in cyanobacteria which can enter into symbiosis with plants. These studies reveal some dissimilarities in the organisation of nif genes and raise questions as to the genetic control of nitrogen fixation in symbiosis.


2006 ◽  
Vol 188 (10) ◽  
pp. 3507-3515 ◽  
Author(s):  
Alycia N. Bittner ◽  
Valerie Oke

ABSTRACT Among the rhizobia that establish nitrogen-fixing nodules on the roots of host plants, many contain multiple copies of genes encoding the sigma factor RpoH and the chaperone GroEL/GroES. In Sinorhizobium meliloti there are two rpoH genes, four groESL operons, and one groEL gene. rpoH1 mutants are defective for growth at high temperature and form ineffective nodules, rpoH1 rpoH2 double mutants are unable to form nodules, and groESL1 mutants form ineffective nodules. To explore the roles of RpoH1 and RpoH2, we identified mutants that suppress both the growth and nodulation defects. These mutants do not suppress the nitrogen fixation defect. This implies that the functions of RpoH1 during growth and RpoH1/RpoH2 during the initiation of symbiosis are similar but that there is a different function of RpoH1 needed later during symbiosis. We showed that, unlike in Escherichia coli, overexpression of groESL is not sufficient to bypass any of the RpoH defects. Under free-living conditions, we determined that RpoH2 does not control expression of the groE genes, and RpoH1 only controls expression of groESL5. Finally, we completed the series of groE mutants by constructing groESL3 and groEL4 mutants and demonstrated that they do not display symbiotic defects. Therefore, the only groESL operon required by itself for symbiosis is groESL1. Taken together, these results suggest that GroEL/GroES production alone cannot explain the requirements for RpoH1 and RpoH2 in S. meliloti and that there must be other crucial targets.


2009 ◽  
Vol 191 (8) ◽  
pp. 2593-2600 ◽  
Author(s):  
Chrysanthi Kalloniati ◽  
Daniela Tsikou ◽  
Vasiliki Lampiri ◽  
Mariangela N. Fotelli ◽  
Heinz Rennenberg ◽  
...  

ABSTRACT Carbonic anhydrase (CA) (EC 4.2.1.1) is a widespread enzyme catalyzing the reversible hydration of CO2 to bicarbonate, a reaction that participates in many biochemical and physiological processes. Mesorhizobium loti, the microsymbiont of the model legume Lotus japonicus, possesses on the symbiosis island a gene (msi040) encoding an α-type CA homologue, annotated as CAA1. In the present work, the CAA1 open reading frame from M. loti strain R7A was cloned, expressed, and biochemically characterized, and it was proven to be an active α-CA. The biochemical and physiological roles of the CAA1 gene in free-living and symbiotic rhizobia were examined by using an M. loti R7A disruption mutant strain. Our analysis revealed that CAA1 is expressed in both nitrogen-fixing bacteroids and free-living bacteria during growth in batch cultures, where gene expression was induced by increased medium pH. L. japonicus plants inoculated with the CAA1 mutant strain showed no differences in top-plant traits and nutritional status but consistently formed a higher number of nodules exhibiting higher fresh weight, N content, nitrogenase activity, and δ13C abundance. Based on these results, we propose that although CAA1 is not essential for nodule development and symbiotic nitrogen fixation, it may participate in an auxiliary mechanism that buffers the bacteroid periplasm, creating an environment favorable for NH3 protonation, thus facilitating its diffusion and transport to the plant. In addition, changes in the nodule δ13C abundance suggest the recycling of at least part of the HCO3 − produced by CAA1.


1976 ◽  
Vol 22 (7) ◽  
pp. 942-948 ◽  
Author(s):  
M. C. Cadmus ◽  
S. P. Rogovin ◽  
K. A. Burton ◽  
J. E. Pittsley ◽  
C. A. Knutson ◽  
...  

Stock cultures of Xanthomonas campestris NRRL B-1459 require special attention to maintenance and propagation to assure consistent production in good yields of the extracellular polysaccharide xanthan. Under customary conditions of propagative maintenance on agar slants, variant colony types develop that are smaller in size than the normal type. The rate of regression of the normal to the variant forms was diminished when the D-glucose content of the stock medium was sufficient to avoid depletion during storage and when transfer to fresh medium was reduced to 14-day intervals. Under conditions for polysaccharide production, the normal large-colony type gives crude culture liquors after 48 h of 7000 centipoise (cp) viscosity; the predominant variant form gives only 4000 cp. On the basis of 2.1% initial D-glucose, biopolymer yields for the normal and variant strains were 62 and 43%, respectively. Polysaccharide produced by the variant (small-colony type) differs adversely in solution properties from that of the parent strain (large-colony type); it differs also in its lower content of pyruvic acid and O-acetyl substituents. The molar ratios of constituent sugars (D-glucose, D-mannose, and D-glucuronic acid), however, were identical in polysaccharides with the normal and variant strains. Exclusion of colonial variants from fermentations is prerequisite to production of xanthan optimum in properties and yield.


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