Lipopolysaccharide and protein patterns of Rhizobium sp. (Galega)

Lipsanen, Päivi

doi:10.1016/0378-1097(89)90061-x

Cited by 12 publications

(21 citation statements)

References 10 publications

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“…A Bradyrhizobium japonicum 110 Lps-derivative fails to elicit any visible nodule development on soybeans (21). On the other hand, LPS 0 antigen may not be required at all in symbiosis by Rhizobium galegae and Rhizobium meliloti SU47 (9,15). Lps mutants of R. leguminosarum CFN42 induce nodules on beans in which infection threads are initiated but usually abort within the root hair cell (8,19).…”

Section: Discussionmentioning

confidence: 99%

Expression of Rhizobium leguminosarum CFN42 genes for lipopolysaccharide in strains derived from different R. leguminosarum soil isolates

et al. 1990

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Two mutant derivatives of Rhizobium leguminosarum ANU843 defective in lipopolysaccharide (LPS) were isolated. The LPSs of both mutants lacked 0 antigen and some sugar residues of the LPS core oligosaccharides. Genetic regions previously cloned from another Rhizobium leguminosarum wild-type isolate, strain CFN42, were used to complement these mutants. One mutant was complemented to give LPS that was apparently identical to the LPS of strain ANU843 in antigenicity, electrophoretic mobility, and sugar composition. The other mutant was complemented by a second CFN42 lps genetic region. In this case the resulting LPS contained 0-antigen sugars characteristic of donor strain CFN42 and reacted weakly with antiserum against CFN42 cells, but did not react detectably with antiserum against ANU843 cells. Therefore, one of the CFN42 lps genetic regions specifies a function that is conserved between the two R. leguminosarum wild-type isolates, whereas the other region, at least in part, specifies a strain-specific LPS structure. Transfer of these two genetic regions into wild-type strains derived from R. leguminosarum ANU843 and 128C53 gave results consistent with this conclusion. The mutants derived from strain ANU843 elicited incompletely developed clover nodules that exhibited low bacterial populations and very low nitrogenase activity. Both mutants elicited normally developed, nitrogen-fixing clover nodules when they carried CFN42 Ips DNA that permitted synthesis of 0-antigen-containing LPS, regardless of whether the 0 antigen was the one originally made by strain ANU843.Leguminous plants and bacteria of the genera Rhizobium and Bradyrhizobium enter into a symbiosis characterized by nitrogen-fixing root nodules. Early steps in development of the symbiosis involve mutual recognition of plant and bacteria, induction of cortical cell division, and (in most cases) invasion of root hairs by way of infection threads. As gram-negative eubacteria, rhizobia have an outer membrane containing lipopolysaccharide (LPS). LPS is composed of two regions: lipid A, which anchors the LPS molecule in the outer leaflet of the outer membrane, and a polysaccharide portion, which projects into the environment. Being a major cell surface molecule, LPS is likely to play a role in early symbiotic events.The polysaccharide of Rhizobium leguminosarum LPS consists of a variable portion and a portion whose composition is conserved among wild-type isolates that have been studied previously (4, 6, 7). The conserved portion is released from the LPS by mild acid hydrolysis as two oligosaccharides that together are composed of galactose, mannose, galacturonic acid, and at the reducing ends, 3-deoxy-Dmanno-octulosonic acid in a 1:1:3:2 molar ratio (4, 5, 14a). The variable portion is found in a longer polysaccharide that is released by mild acid hydrolysis. Its composition varies from strain to strain (4, 26). The variable portion carries the dominant antigenic determinants of wild-type strains and is referred to as the 0 antigen, in keeping with the termin...

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Section: Discussionmentioning

confidence: 99%

Expression of Rhizobium leguminosarum CFN42 genes for lipopolysaccharide in strains derived from different R. leguminosarum soil isolates

et al. 1990

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“…So far there is only one report on the characterization of LPSs from R. galegae (20). Various strains have been characterized by comparison of their LPS gel patterns and the whole cell protein patterns.…”

Section: Methodsmentioning

confidence: 99%

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae

et al. 1991

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Lipopolysaccharides (LPSs) isolated from several strains of Rhizobium, Bradyrhizobium, Agrobacterium, and Azorhizobium were screened for the presence of 27-hydroxyoctacosanoic acid. The LPSs from all strains, with the exception of Azorhizobium caulinodans, contained various amounts of this long-chain hydroxy fatty acid in the lipid A fractions. Analysis of the lipid A sugars revealed three types of backbones: those containing glucosamine (as found in Rhizobium meliloti and RhizobiumfrediO), those containing glucosamine and galacturonic acid (as found in Rhizobium leguminosarum bv. phaseoli, trifolii, and viciae), and those containing 2,3-diamino-2,3-dideoxyglucose either alone or in combination with glucosamine (as found in Bradyrhizobium japonicum and Bradyrhizobium sp.[Lupinus] strain DSM 30140). The distribution of 27-hydroxyoctacosanoic acid as well as analysis of lipid A backbone sugars revealed the taxonomic relatedness of various strains of the Rhizobiaceae.Bacteria belonging to the family Rhizobiaceae are gram negative and are able to form nitrogen-fixing symbiotic relationships with legume plants. There are three distinct genera: the symbiotic nitrogen-fixing Rhizobium and Bradyrhizobium spp. and the plant pathogenic Agrobacterium spp. Quite recently a new genus, so far comprising only the stem-nodulating nitrogen-fixing species Azorhizobium caulinodans, was defined (13). Of these genera, the species of Rhizobium are taxonomically closely related and show genetic similarities to the genus Agrobacterium as evidenced by 16S rRNA homology studies (12). On the other hand, the slowly growing species of Bradyrhizobium are rather distantly related to the other two genera as revealed by their low SAB values determined by DNA-rRNA hybridization studies (1). In addition to the nucleotide sequence homology studies, differentiation of various members of the Rhizobiaceae has been attempted by several chemotaxonomic approaches such as cellular fatty acid analysis (21, 31), polyacrylamide gel electrophoresis of cellular proteins (19), and composition analysis of extracellular gum (26). However, results of these studies were not sufficient to adequately distinguish between members of the Rhizobiaceae. More recently, the backbone sugar composition of lipid A fractions of lipopolysaccharide (LPS) has been used as a taxonomic marker for recognition and relatedness of various nonsulfur bacteria (23). Therefore, in this study, the lipid A fractions from rhizobial LPSs were examined to see whether they represented a marker for determining the relatedness of these bacteria.The surface polysaccharides, including the LPS, of strains of Rhizobium have been hypothesized to be involved in the molecular mechanisms of symbiotic infection (5). In an attempt to elucidate the structure of LPS from rhizobial strains, an unusual very-long-chain hydroxy fatty acid, 27-hydroxyoctacosanoic acid (27-OH-28:0), was discovered to be the major fatty acid constituent of the lipid A region (15). More recently, we have also identified this long-cha...

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“…Features that distinguish goat's rue rhizobial strains from other bacteria within the genus Rhizobium are DNA homology (13,28), rRNA-DNA homology (8), bacteriophage typing (11,13), lipopolysaccharide and protein patterns (18), and metabolic properties (14). For example, the mean relative DNA homology between 10 strains nodulating Galega sp.…”

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confidence: 99%

Restriction fragment length polymorphism analysis of Rhizobium galegae strains

Kaijalainen

Lindström

1989

J Bacteriol

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Total DNA of various Rhizobium galegae strains representing different geographical origins, and taxonomic divergence was digested with three restriction enzymes separately, Southern blotted, and hybridized with six heterologous probes. The sequence divergences for different pairwise comparisons were calculated from proportions of conserved hybridizing fragments. The unweighted pair group method was used to group the strains. The symbiotic common nod and niJHDK probes used were highly conserved and grouped the strains according to the host plant, Galega orientalis or G. officinalis. The grouping derived from combined data of the constitutive hemA, gInA, ntrC, and recA probes was similar to that obtained in total DNA-DNA hybridization experiments. The constitutive probes grouped the strains in a different order than did the symbiotic probes, a result that may reflect interstrain transfer of symbiotic sequences in the course of evolution.Fast-growing rhizobia nodulating the legume species Galega officinalis and G. orientalis (goat's rue) have recently received attention because of the potential agronomic importance of their host plants as perennial nitrogen-fixing forage crops for temperate regions (10,12,15,27). Features that distinguish goat's rue rhizobial strains from other bacteria within the genus Rhizobium are DNA homology (13, 28), rRNA-DNA homology (8), bacteriophage typing (11, 13), lipopolysaccharide and protein patterns (18), and metabolic properties (14). For example, the mean relative DNA homology between 10 strains nodulating Galega sp. with reference DNA from strain HAMBI (The Culture Collection at the Department of Microbiology, University of Helsinki) 540, which forms effective (nitrogen-fixing) nodules on G. orientalis, was 77 ± 9%, compared with 19 ± 16% homology between HAMBI 540 and other rhizobia and bradyrhizobia (13). The Galega rhizobia are also very host specific, nodulating only their own host plants. Similarly, rhizobia from other species do not normally infect the Galega plants (13,16). The Galega rhizobia infect their host plants through the root hairs and form nodules with an apical meristem. Strains isolated from G. orientalis form effective nodules on that plant but ineffective nodules on G. officinalis and vice versa (13,17). It has recently been suggested that the Galega rhizobia form their own species, Rhizobium galegae (10).In this work, we studied the genetic relatedness between R. galegae strains representing different geographic origins and taxonomic divergence observed in earlier studies. Four strains of G. orientalis and seven strains of G. officinalis were included. Total DNA isolated from the strains was digested with restriction enzymes and hybridized separately to sequences carrying common nod genes of Rhizobium meliloti, nifHDK, hemA, and recA genes of R. meliloti, and glnA and ntrC genes of Azorhizobium caulinodans. Restriction fragment length polymorphism of fragments hybridizing with the DNAs was used to calculate the sequence divergences between the strains; on th...

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Lipopolysaccharide and protein patterns of Rhizobium sp. (Galega)

Cited by 12 publications

References 10 publications

Expression of Rhizobium leguminosarum CFN42 genes for lipopolysaccharide in strains derived from different R. leguminosarum soil isolates

Expression of Rhizobium leguminosarum CFN42 genes for lipopolysaccharide in strains derived from different R. leguminosarum soil isolates

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae

Restriction fragment length polymorphism analysis of Rhizobium galegae strains

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