• Korean Journal of Microbiology
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• v.26 no.3
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• pp.215-222
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• 1988

The effect of ammonia and glutamine on nitrogenase activity of Rhodopseudomonas sphaeroides was examined. The nitrogenase activity of this strain was inhibited by ammonia and glutamine. When ammonia and glutamine were exhausted, nitrogenase activity promptly resumed at its original rate. Methionine sulfoximine (MSX), irreversible glutamine synthetase (GS) inhibitor, is a structural analogue of glutamate. MSX was used in order to know whether the nitrogenase activity was inhibited by ammonia and glutamine directly or not. The ability of MSX to prevent nitrogenase switch-off by ammonia was found to be dependent upon the phase of culture. When the cells were sampled after 12 hour culture, $500{\mu}M$ MSX would not prevent the nitrogenase switch-off by ammonia. Twenty one percents of GS actibity was inhibited by $500{\mu}M$ of MSX and concentration of released ammonia decreased. But nitrogenase activiy was still inhibited by ammonia. However, nitrogenase switch-off after 20 hours would be prevented by $100{\mu}M$ of MSX. On the other hand, GS activity was ingibited completely by $100{\mu}M$ MSX and concentration of released ammonia somewhat increased. But nitrogenase activity was not inhibited. The data indicated that the inhibition of in vivo nitrogenase actibity of Rp. sphaeroides by ammonia seemed to be mediated by products of ammonia assimilation rather than by ammonia itself.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.211-216
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• 1983

Rhodopseudomonas sphaeroides K-7 evolves large quantities of molecular hydrogen under anaerobic and light illuminated conditions in the presence of utilizable organic compounds as electron donors. Photoevolution of molecular hydrogen was strictly dependent on light as the activity of nitrogenase in this organism. Both of these were inhibited to the nearly same extent at varying concentrations of ammonium ion which also depressed nitrogenase synthesis. In the reaction mixtures devoid of molybdenum ion which is known as the component of nitrogenase, hydrogen evolution also decreased similarly like nitrogenase activity. Photoevolution of molecular hydrogen appeared to have no relationship with hydrogenase activity and bacteriocholophyll content and it was markedly inhibited under the atmosphere of $C_2$H$_2$, $N_2$ or $O_2$. The results strongly indicate that hydrogen evolution by R. sphaeroides K-7 might be catalyzed by nitrogenase. Both hydrogen evolution and nitrogenase activity were largely influenced by the nutritional history of the resting cells. From which we propose that glutamate might play an important role in the regulation of nigrogenase activity in vivo.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.5
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• pp.361-368
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• 2003

The responses of five varieties and three cultivars of pea (Pisum sativum) to Rhizobium inoculation on nodulation, growth, nitrogenase activity, dry matter production and N uptake were investigated. The pea varieties were IPSA Motorshuti-l, IPSA Motorshuti-2, IPSA Motorshuti-3, BARI Motorshuti-l, BARI Motorshuti-2 and the cultivars were 063, Local small and Local white. Fifty percent seeds of each pea variety/cultivar were inoculated with a mixture of Rhizobium inoculants at rate of 15g/kg seed and the remaining fifty percent seeds were kept uninoculated. The plants inoculated with Rhizobium inoculant significantly increased nodulation, growth, nitrogenase activity, dry matter production and N uptake. Among the varieties/cultivars, BARI Motorshuti-l performed best in almost all parameters including nitrogenase activity of root nodule bacteria of the crop. There were positive correlations among the number and dry weight of nodules (r=$0.987^{**}$, $0.909^{**}$), nitrogenase activity of root nodule bacteria (r=$0.944^{**}$, $0.882^{**}$), dry weight of shoot (r=$0.787^{**}$, $0.952^{**}$), N content (r=$0.594^{**}$, $0.605^{**}$) and N uptake (r=$0.784^{**}$, $0.922^{**}$) by shoot both at flowering and pod filling stages of the crop, respectively. It was concluded that BARI Motorshuti-l in symbiotic association with Rhizobium inoculant performed best in recording nitrogenase activity, dry matter production and N uptake by pea.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.5
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• pp.355-360
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• 2003

The responses of six mungbean [Vigna radiata (L.) Wilczek] varieties to Rhizobium inoculation on number and dry weight of nodules, nitrogenase activity of root nodule bacteria, dry weight of shoot and root, nitrogen content, and uptake by shoot were investigated. The mungbean varieties were BARI Mung-2, BARI Mung-3, BARI Mung-4, BARI Mung-5, BINA Moog-2, and BU Mung-1. Two-third seeds of each variety were inoculated with Rhizobium inoculant and the remaining one-third seeds were kept uninoculated. Rhizobium strains TAL 169 and TAL 441 were used for inoculation of seeds. Inoculation of seeds with Rhizobium strains significantly increased nodulation, nitrogenases activity, dry matter production, nitrogen content, and uptake by shoot of the crop compared to uninoculated control. There was positive correlation among the number and dry weight of nodules, nitrogenase activity, dry weight of shoot and root, nitrogen content, and uptake by shoot of the crop. It was concluded that BARI Mung-4 in association with Rhizobium strain TAL 169 performed best in recording nodulation, nitrogenase activity, dry matter production, and nitrogen uptake by shoot of mungbean.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.113-118
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• 1986

Optimum temperature and pH of glutamine synthetase activity (E.C. 3.6.1.2.) of R. sphaeroides D-230 was $35^{\circ}C$ and 6.8, respectively. The adenylated state of GS in R. sphaeroides D-230 was stabilized by addition of 0.2mg/ml of cethyltrimethylammoniumbromide. Valine, histidine, proline, isoleucine, and lysine were good nitrogen source for the growth of R. sphaeroides D-230. The growth of R. sphaeroides D-230 in $N_2,\;NaNO_3\;or\;NH_4Cl$ as sole nitrogen source was lower than in any otherculture conditions. GS activity was inhibited, more or less, by various amino acid. THe relative inhibition rate of the enzyme by added 7mM arginine, $NH_4Cl,\;N_2,\;and\;NaNO_3$ was 63.8%, 26.79%, 6.24%, and 10.64%, drespectively. THe hydrogen evolution of R. sphaeroides D-230 grown in N-limited media was inhibited by 0.1mM MSX, irreversible GS inhibitor. GS activity was completely inhibited by 1.0mM MSX but ammonia released maximally at the same concentration of MSX. Ammonia release by added MSX was increased up to 1.0mM MS, but decreased above 1.0mM MSX. It is probably due to inhibition of nitrogenase actixity by MSX. Nitrogenase activity was not inhibited at low concentration of MSX. These results suggests that the inhibition of nitrogenase activity by ammonia is mediated by products of ammonia assimilation rather than by ammonia itself.

• Korean Journal of Soil Science and Fertilizer
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• v.12 no.1
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• pp.43-46
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• 1979

Curing the studies on the mechanism of the nodule formation and the infection of rhizobia using the continously cultured plant cell tissues, it was found that some calluses possess high nitrogen fixation activity. This experiment was conducted to know the differences among the calluses and the Rhizobia. The results obtained were as follows; 1) In a single inoculation the nitrogenase activity of soybean cell cultures-rhizobium was moderately higher than non-leguminous cell cultures, however, in the mixing inoculation that was reversely found. 2) Host factor, which was characteristics for the nodule formation and the nitrogenase activity, was not appeared generally in the plant cell cultures except for Kuamusume and Toyozuzu in soybean, and Datura in non-leguminous cell cultures. 3) In the 012 rhizoblium cultured on soybean cell cultures and in the 010, 023 and 024 rhizobia cultured on non-leguminous cell cultures the nitrogenase activity higher than the others.

• The Korean Journal of Food And Nutrition
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• v.5 no.2
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• pp.137-143
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• 1992

As a research for developing biofertilizers, Klebsiella oxytoca, an associative nitrogen fixer in the rhizosphere of rice plant in the soil of paddy field, was subjected to molecular breeding. The results obtained were as followings. 1). By transforming pbIC71A, Nif A overproducing plasmid, into Klebsiella oxytoca NGl3, Klebsiell6f oxytoca SH3l, and Klebsiella oxytoca SH161, nitrogenase activities in the absence of nitrogen source in the medium were increased 6.4, 17.2, and 13.5 times, respectively, in comparison with the parent strains. 2). Nitrogenase activity of Klebgiella oxytoca NGl3, Klebsiella oxytoca SH3l, and Klebsiella oxytoca SH161 was completely repressed In the presence of 15mM NH4+. But, nitrogenase activities of Klebsiella oxytoca NGl3/PMC71A, Klebsiella oxytoca SH3l /PMC71A, and Klebsiella oxytoca SH 161/pMC714 harboring PMC71A, were 13.7%, 7.7%, and 6.2% of the nitrogenase activities in the absence of nitrogen source in the medium, respectively.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.18-24
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• 2003

For researching nitrogen-fixing bacteria associated with gramineous crops, we collected growing roots of rices, wheats, oats, barleys, ryes, and maizes at 19 sites of southern Korean peninsula. Endophytes and free living bacteria were isolated from those crop roots. Sixty-three isolates were classified on the basis of different morphology, size, color, host of colony, and the 16S rDNAs sequence. The analyses of PCR amplification for nifH gene and nitrogenase activity assay, revealed that all isolates contained nitrogen-fixing abilities. In addition, most of them have cellulase activity which is one of the common features of endophytic bacteria from plant.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.304-309
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• 1989

The effects of simazine [2-chloro-4, 6-bis(methylamino)-s-triazine] on in vivo nitrogenase, glutamine synthetase, glutamate synthase, and glutamate dehydrogenase which are important in nitrogen assimilation of Rhodopseudomonas sphaeroides were investigated. Simazine inhibited the synthesis of nitrogenase and glutamine synthetase. The activity of glutamine synthetase in crude extracts of Rhodopseudomonas sphaeroides is less inhibited by simazine than that of glutamate synthase and glutamate dehydrogenase. These results suggest the possibility that simazine inhibits photosynthetic activity and so inhibits the synthesis of nitrogenase and glutamine synthetase in Rhodopseudomonas sphaeroides.

• Applied Biological Chemistry
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• v.41 no.1
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• pp.53-59
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• 1998

The Rhizobium and Azospirillum spp. were isolated from the root nodules of several leguminous plants and rhizosphere of various paddy rice varieties. The growth of the nitrogen-fixing strains isolated was largely inhibited in yeast extract-mannitol medium (AMA) containing 0.6 M NaCl. In response to osmotic stress, the nitrogen-fixing strains accumulate intracellular free glutamate. The growth and nitrogenase activity of Rhizobium and Azospirillum were increased by addition of osmoprotectants such as proline, glycine betaine, and glutamate during salt stress. Glycine betaine was the most effective among exogenous osmoprotectants tested. In the absence of sodium chloride, nitrogenase activity seem to be slightly decreased by the presence of the proline or glycine betaine. These results revealed that nitrogenase activity was repressed by fixed nitrogens such as proline or glycine betaine.