Growth of nodule bacteria in soil]

As was found with the aid of capillary microscopy, the nodule bacteria of pea and lupine in soil are encountered as rods, bacteroids, and cocci. The rod-like cells form bacteroids not only in the nodules but also outside, in soil. The bacteroids are viable and later reproduce the coccoid cells (arthrospores). The rods and bacteroids prevail in soil during flowering of the legumes, and the cocci are predominant at the end of vegetation.     

Siderophore and organic acid production in root nodule bacteria

Nineteen strains of root nodule bacteria were grown under various iron regimes (0.1, 1.0 and 20 M added iron) and tested for catechol and hydroxamate siderophore production and the excretion of malate and citrate. The growth response of the strains to iron differed markedly. For 12 strains (Bradyrhizobium strains NC92B and 32H1, B. japonicum USDA110 and CB1809, B. lupini WU8, cowpea Rhizobium NGR234, Rhizobium meliloti strains U45 and CC169, Rhizobium leguminosarum bv viciae WU235 and Rhizobium leguminosarum bv trifolii strains TA1, T1 and WU95) the mean generation time showed no variation with the 200-fold increase in iron concentration. In contrast, in Bradyrhizobium strains NC921, CB756 and TAL1000, B. japonicum strain 61A76 and R. leguminosarum bv viciae MNF300 there was a 2–5 fold decrease in growth rate at low iron. R. meliloti strains WSM419 and WSM540 showed decreased growth at high iron.All strains of root nodule bacteria tested gave a positive CAS (chrome azurol S) assay for siderophore production. No catechol-type siderophores were found in any strain, and only R. leguminosarum bv trifolii T1 and bv viciae WU235 produced hydroxamate under low iron (0.1 and 1.0 M added iron).Malate was excreted by all strains grown under all iron regimes. Citrate was excreted by B. japonicum USDA110 and B. lupini WU8 in all iron concentrations, while Bradyrhizobium TAL1000, R. leguminosarum bv viciae MNF300 and B. japonicum 61A76 only produced citrate under low iron (0.1 and/or 1.0 M added iron) during the stationary phase of growth.Abbreviations CAS chrome azurol S - HDTMA hexadecyltrime-thylammonium bromide     

Growth-promoting effect of chloro-substituted aliphatic acids in root nodule bacteria

Summary Trichloro-, dichloro- and monochloroacetic and -dichloro-, -monochloro-and -monochloropropionic acids as sodium salts in 0.10–1.0mm concentration accelerated the growth of a strain of Rhizobium leguminosarum in a liquid synthetic medium with biotin as sole accessory growth factor. The strongest effect was shown by -monochloropropionate. Concentrations of 10mm and above were inhibitory. -Monochlorobutyrate did not accelerate growth.Ca-pantothenate and its precursor -alanine strongly improved growth in concentrations of 0.10 to 1.0 p.p.m. (0.001–0.011mm -alanine) and masked the effect of the chloro-substituted organic acids which seemed to function as more or less inferior substitutes for -alanine.The effect of -alanine was approximately ten times greater than that of -monochloropropionate which possibly owed its relatively strong activity to its close structure analogy with -alanine.Two other strains of rhizobia showed a moderate response to trichloroacetate and dichloropropionate, while others failed to do so and grew abundantly with biotin alone.     

The life of root nodule bacteria in the acidic underground

Abstract The effects of low pH on the growth and survival of root nodule bacteria are important in determining the success of legume inoculation in practical agriculture. The mechanisms for maintenance of intracellular pH (pH_i) are crucial, but little is yet known about them in root nodule bacteria. Like other Gram-negative bacteria, root nodule bacteria show an adaptive acid tolerance response, with growth at moderately acidic conditions protecting against an extreme acid shock. Variations in acid tolerance within species of root nodule bacteria imply a genetic basis to low pH tolerance and studies of acid-sensitive mutants suggest that as many as 20 genes could be involved. A general model for acid tolerance in root nodule bacteria is presented.     

Novel root nodule bacteria belonging to the genus Caulobacter

Aims: Aim of this study is to determine the genetic variation of rhizobia associated with horse gram [Macrotyloma uniflorum (Lam.) Verdc.] plants grown in different regions of Andhra Pradesh, India. Methods and Results: Four representative isolates having most representative characters from the previous characterization were selected for 16S rRNA sequence. The sequences were submitted to the NCBI GenBank and Ribosomal Database Project (RDP). The isolates HGR‐4, 6 and 13 showed more than 99% homology between them and they were grouped with Rhizobium reference strains where as the isolate HGR‐25 showed 87·1, 87·4 and 87·2% homology with the isolates HGR‐4, 6 and 13, respectively, and were grouped with reference strains for Caulobacter. The nodulation ability of these isolates on horse gram was confirmed by inoculation tests. Conclusions: The isolate HGR‐25 was identified as Caulobacter isolated from the plants growing in soil samples collected from Khareemnagar district, Andhra Pradesh, India. Inoculation tests revealed that Caulobacter formed nodules on horse gram. It was also confirmed by RDP. Significance and Impact of the Study: This is the first report that a legume was nodulated by a member of the genus Caulobacter, which belongs to the family Caulobacteriaceae in the order Caulobacterales of Alphaproteobacteria.     

Fast-growing root nodule bacteria produce a novel polyamine, aminobutylhomospermidine

Polyamines in various root nodule bacteria including Bradyrhizobium japonicum, Rhizobium fredii, R. leguminosarum, R. meliloti and R. loti were identified by capillary gas chromatography. Homospermidine was the polyamine present in highest concentration in all the rhizobia tested. In addition to putrescine and homospermidine, fast-growing type of rhizobial cells contained a novel polyamine, aminobutylhomospermidine, NH2(CH2)4NH(CH2)4NH(CH2)4NH2. The unusual tetraamine was not found in the cells of slow-growing type of rhizobia throughout their growth period, indicating a difference in polyamine metabolism between fast-growing type and slow-growing type of root nodule bacteria.     

Susceptibility to hydrogen peroxide and catalase activity of root nodule bacteria

The root nodule bacteria (free-living cells) tested had higher susceptibility to hydrogen peroxide (H2O2) than the other genera of aerobic or facultative anaerobic bacteria tested. The catalase activities tended to have a positive correlation with H2O2 resistance among all bacteria tested. Addition of a catalase inhibitor such as 3-amino-1, 2, 4-triazole increased the susceptibility to H2O2. These results suggest that the lower catalase activity brings about the higher susceptibility of root nodule bacteria to H2O2. Root nodule bacteria seemed to have two or three catalase isozymes during growth and their catalase activities were higher in log phase than in stationary phase, contrary to other genera of bacteria tested.     

The adaptive acid tolerance response in root nodule bacteria and Escherichia coli

Root nodule bacteria and Escherichia coli show an adaptive acid tolerance response when grown under mildly acidic conditions. This is defined in terms of the rate of cell death upon exposure to acid shock at pH 3.0 and expressed in terms of a decimal reduction time, D. The D values varied with the strain and the pH of the culture medium. Early exponential phase cells of three strains of Rhizobium leguminosarum (WU95, 3001 and WSM710) had D values of 1, 6 and 5 min respectively when grown at pH 7.0; and D values of 5, 20 and 12 min respectively when grown at pH 5.0. Exponential phase cells of Rhizobium tropici UMR1899, Bradyrhizobium japonicum USDA110 and peanut Bradyrhizobium sp. NC92 were more tolerant with D values of 31, 35 and 42 min when grown at pH 7.0; and 56, 86 and 68 min when grown at pH 5.0. Cells of E. coli UB1301 in early exponential phase at pH 7.0 had a D value of 16 min, whereas at pH 5.0 it was 76 min. Stationary phase cells of R. leguminosarum and E. coli were more tolerant (D values usually 2 to 5-fold higher) than those in exponential phase. Cells of R. leguminosarum bv. trifolii 3001 or E. coli UB1301 transferred from cultures at pH. 7.0 to medium at pH 5.0 grew immediately and induced the acid tolerance response within one generation. This was prevented by the addition of chloramphenicol. Acidadapted cells of Rhizobium leguminosarum bv. trifolii WU95 and 3001; or E. coli UB1301, M3503 and M3504 were as sensitive to UV light as those grown at neutral pH.