Effect of Yeast Extract Concentration on Viability and Cell Distortion in <i>Rhizobium</i> spp.

Skinner, F. A.; Roughley, R. J.; Chandler, Muriel R.

doi:10.1111/j.1365-2672.1977.tb00753.x

Cited by 25 publications

(15 citation statements)

References 16 publications

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“…Glycine toxicity in bacteria is a well-documented phenomenon and has been linked to a 20 to 40% decrease in the extent of cross-linking of the peptidoglycan and significant changes in cell morphology, including elongation, spheroplast formation, and rounding of rod-shaped cells (26). Yeast extract or glycine induces significant changes in cell morphology in some strains of Rhizobium spp., resulting in alteration of the typical rodshaped cells into enlarged, distorted, pleomorphic shapes and spheroplasts that sometimes resemble bacteroid cells (46,47). The RL3499-RL3502 mutants had similarly altered cell shape in the presence of glycine, providing further evidence for cell wall alterations in these mutants.…”

Section: Discussionmentioning

confidence: 72%

“…Earlier research studying rhizobial growth has documented sensitivity to yeast extract and peptide sources such as hydrolyzed casein (46,47). In some reports this sensitivity was attributed to the presence of the amino acid glycine (46).…”

Section: Resultsmentioning

confidence: 99%

“…Alterations in the cell morphology of R. leguminosarum when grown in the presence of yeast extract and glycine have been previously observed (46,47). Therefore, the cell morphologies of the SM1 and nonpolar mutants were examined using light microscopy.…”

Section: Resultsmentioning

confidence: 99%

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Mutation of a Broadly Conserved Operon (RL3499-RL3502) from Rhizobium leguminosarum Biovar viciae Causes Defects in Cell Morphology and Envelope Integrity

et al. 2011

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The bacterial cell envelope is of critical importance to the function and survival of the cell; it acts as a barrier against harmful toxins while allowing the flow of nutrients into the cell. It also serves as a point of physical contact between a bacterial cell and its host. Hence, the cell envelope of Rhizobium leguminosarum is critical to cell survival under both free-living and symbiotic conditions. Transposon mutagenesis of R. leguminosarum strain 3841 followed by a screen to isolate mutants with defective cell envelopes led to the identification of a novel conserved operon (RL3499-RL3502) consisting of a putative moxR-like AAA ؉ ATPase, a hypothetical protein with a domain of unknown function (designated domain of unknown function 58), and two hypothetical transmembrane proteins. Mutation of genes within this operon resulted in increased sensitivity to membranedisruptive agents such as detergents, hydrophobic antibiotics, and alkaline pH. On minimal media, the mutants retain their rod shape but are roughly 3 times larger than the wild type. On media containing glycine or peptides such as yeast extract, the mutants form large, distorted spheres and are incapable of sustained growth under these culture conditions. Expression of the operon is maximal during the stationary phase of growth and is reduced in a chvG mutant, indicating a role for this sensor kinase in regulation of the operon. Our findings provide the first functional insight into these genes of unknown function, suggesting a possible role in cell envelope development in Rhizobium leguminosarum. Given the broad conservation of these genes among the Alphaproteobacteria, the results of this study may also provide insight into the physiological role of these genes in other Alphaproteobacteria, including the animal pathogen Brucella.

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Section: Discussionmentioning

confidence: 72%

Section: Resultsmentioning

confidence: 99%

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Mutation of a Broadly Conserved Operon (RL3499-RL3502) from Rhizobium leguminosarum Biovar viciae Causes Defects in Cell Morphology and Envelope Integrity

et al. 2011

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“…Methyl blue reduction was observed by adding 1 drop of 1% methyl blue to a test tube of inoculated milk after 7 days of incubation at 28°C and then incubating the preparation at 37°C for 1 h; disappearance of the blue color was regarded as a positive reaction. Nile blue reduction was detected by using the method of Skinner et al (16). Urease was detected by the methods of Smibert and Krieg (17), whereas the catalase, phenyalaninase, oxidase, peroxidase, and cytochrome oxidase tests were those described by Skerman (15).…”

Section: Methodsmentioning

confidence: 99%

Numerical Taxonomic Study of Fast-Growing Soybean Rhizobia and a Proposal that Rhizobium fredii Be Assigned to Sinorhizobium gen. nov.

Chen

Yan

1988

International Journal of Systematic Bacteriology

306

181

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A total of 33 strains of fast-growing soybean rhizobia isolated from soil and soybean nodules collected in China and 25 strains belonging to the genera Rhizobium, Bradyrhizobium, and Agrobacteriurn were compared by numerical taxonomic techniques, using 240 different characters, Our results indicated that all of the strains of fast-growing soybean rhizobia which we examined are closely related (guanine-plus-cytosine content, 59.9 to 63.8 mol%) and are separated from Rhizobium and Bradyrhizobium at the generic level. Based on numerical taxonomy, deoxyribonucleic acid (DNA) base ratio determinations, DNA-DNA hybridization data, serological analysis data, the composition of extracellular gum, bacteriophage typing data, and soluble protein patterns, we propose that the fast-growing soybean rhizobia represent members of a new genus rather than a species of Rhizobium (Rhizobium fredii); we propose Sinorhizobium gen. nov. as an appropriate generic name. (4). Rhizobium includes three species, Rhizobium meliloti,Rhizobium loti, and Rhizobium leguminosarum. Bradyrhizobium has only one species, Bradyrhizobium japonicum, and includes strains that are capable of nodulating lupines, soybeans, and certain other legumes (4).Recently, a new group of fast-growing soybean rhizobia was identified from soil and soybean nodules collected in the People's Republic of China by Chinese and American scientists (6, 25). The taxonomic status of these bacteria was uncertain, but they may represent a transitional group falling between the genera Rhizobium and Bradyrhizobium (4).Scholla and Elkan (13) proposed a new species name for this group, Rhizobium fredii, based mainly on deoxyribonucleic acid (DNA) hybridization comparisons of five strains of these bacteria with representatives of the genera Rhizobium and Brudyrhizobium and on a review of some of the characteristics of 11 strains of fast-growing soybean rhizobia.To extend the current information on the fast-growing soybean bacteria, we performed a numerical taxonomic study of a large number of strains. We propose that they be assembled in a new genus that includes two species. MATERIALS AND METHODSOrganisms. A total of 58 strains were used in this study (Table 1); 33 strains were fast-growing soybean rhizobia isolated from soil and soybean nodules collected in the People's Republic of China, and the other 25 strains were representatives of Rhizobium, ~r u d y r~i z o~i u m , and Agrobacterium.Features. Each of the 58 strains was characterized by determining 240 different coding features. The following features were considered: (i) utilization as sole carbon sources (0.1%) of cellobiose, D-mannose, salicin, ammonium tartrate, casein hydrolysate, fructose, sodium acetate, sorbose, riffinose, D-melezitose monohydrate, hexose 6-phos- (iv) tolerance to dyes (0.1%, wt/vol), such as sudan 1, erythrosin A, picrocarmine, safranine T, orcein, rosanilin, brilliant cresyl blue, gentian violet, bromthymol blue, bromophenol blue, methyl red, bromocresol purple, neutral red, auramine O.B.S, light gree...

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“…*Present address: P. G. Department of Botany, 'Utkal University (Vani Vihar), Orissa, India. 84 A. Misra et al Starch was identified by iodine staining, the presence of polyphosphate and glycogen was assumed on the basis of tests on similar materials (Dart & Chandler 1972;Skinner et al 1977); the presence of leghaemoglobin by pink coloration of the nodules. …”

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A comparative study by electron microscopy of the structures of effective and ineffective nodules ofTrifolium subterraneum induced byRhizobium trifolii andRhizobium leguminosarum

Misra

Chandler

Hepper

1985

World J Microbiol Biotechnol

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IntroductionAn extensive study by light microscopy of effective nodules of Trifolium subterraneum has been published by Roughley et al. (1976a,b), but there has been little electron microscopy of effective subterranean clover nodules, and no detailed study of the ineffective nodules except the brief examination of ineffective bacteroids in this species by Bergersen (1955).Hepper (1978) showed that the infrequent sparse nodulation of red clover (T. pratense) by Rhizobium leguminosarum could be enhanced by selection and breeding. Hepper & Lee (1979) subsequently reported that R. leguminosarum strains 1013 and 1020 nodulated T. subterraneum but showed no nitrogenase activity. This paper describes the structure of effective and ineffective nodules induced on subterranean clover roots by R. trifolii, and the ineffective nodules formed by two stains of R. leguminosarum as revealed by light and electron microscopy. Materials and methodsMedium sized seeds of Trifolium subterraneum, c.v. Mount Barker, were grown as described by Hepper & Lee (1979). Seedlings were inoculated with one of the following strains from the Rothamsted Culture Collection: 1 R. trifolii strain 211 (TA1) -effective. 2 R. trifolii strain 6 -(Coryn) which ineffectively nodulates a range of clover spp., 3 R. leguminosarum strain 1013 -effective on Pisum sativum, Vicia hirsuta and V. sativa, 4 R. leguminosarum strain 1020A -effective on Pisum sativum and Vicia sativa, but ineffective on V. hirsuta. Root maps were drawn daily to record the exact age and position of the nodules as they became visible. Nodules were fixed for both light and electron microscopy at 1, 3, 5, 7, 9 and 14 days, and at 21 days for light microscopy only. Nodules for electron microscopy were fixed, embedded, stained and examined as described by Chandler (1978). Material for light microscopy was fixed in 10% acrolein in tap water and embedded in glycol methacrylate. Sections of 1.5-2.0/am were stained with 0.5% Toluidine blue in benzoate buffer. *Present address: P. G. Department of Botany, 'Utkal University (Vani Vihar), Orissa, India. 84 A. Misra et al. Starch was identified by iodine staining, the presence of polyphosphate and glycogen was assumed on the basis of tests on similar materials (Dart & Chandler 1972;Skinner et al. 1977); the presence of leghaemoglobin by pink coloration of the nodules. Results and discussion Effective nodules, strain 211 (TA1)Nodules on plants inoculated with R. trifolii strain 211 (TA1) were visible 5 days after inoculation and occurred on primary and secondary roots. The five zones defined by Roughley et al. (1976a) were found in mature nodules: the meristem, infection, differentiating, bacteroid, and degeneration zone. Small spherical 1-day old nodules showed only three zones; meristem, infection zone, and a small differentiating zone. These, with the developing vascular trace were enclosed in an endodermis surrounded by a husk comprising two to four layers of cells. The infection zone, which was extensive, showed many branching infection t...

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Effect of Yeast Extract Concentration on Viability and Cell Distortion in Rhizobium spp.

Cited by 25 publications

References 16 publications

Mutation of a Broadly Conserved Operon (RL3499-RL3502) from Rhizobium leguminosarum Biovar viciae Causes Defects in Cell Morphology and Envelope Integrity

Mutation of a Broadly Conserved Operon (RL3499-RL3502) from Rhizobium leguminosarum Biovar viciae Causes Defects in Cell Morphology and Envelope Integrity

Numerical Taxonomic Study of Fast-Growing Soybean Rhizobia and a Proposal that Rhizobium fredii Be Assigned to Sinorhizobium gen. nov.

A comparative study by electron microscopy of the structures of effective and ineffective nodules ofTrifolium subterraneum induced byRhizobium trifolii andRhizobium leguminosarum

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