• Current Research on Agriculture and Life Sciences
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• v.11
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• pp.121-132
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• 1993

This study was carried out to research the effect of the chemotaxis of Bradyrhizobium japonicum KCTC 2422 and its mutant toward soybean root exudate and to elucidate the effect of the lectin of host specificity (Host Recognition) in soybean-Bradyrhizobium symbiosis. The results obtained were as follows: The homogeneities of the purified lectins from soybean and pea seed was ascertained chromatographically and electrophoretically. Gel electrophoresis of soybean lectin in the presence of sodium dodecyl sulfate appeared a single protein band, whereas pea lectin appeared two protein bands. Soybean lectin from 2 cultivars formed immunoprecipitin arcs at same position with anti-soybean lectin rabbit IgG, but pea lectin did not form immunoprecipitin lines with anti-soybean lectin rabbit IgG. Chemotactic responses of KCTC 2422, LPN-100 and LCR-101 toward proline in capillary assays were 3.1, 1.3 and 1.0-fold above background, respectively. The chemotactic responses of KCTC 2422, LPN-100, and LCR-101 toward Paldal crude root exudate in capillary assays were 3.5, 1.4 and 1.4-fold above background, respectively. The present work shows that B. japonicum and its mutants are capable of very different responses toward root exudate fraction. The chemotactic responses of KCTC 2422 was most with neutral fraction, least with anionic fraction and intermediate with cationic fraction. The nitrogenase activity of soybean nodule was shown in 15days after inoculation with LCR-101. However, we couldn't find out the nodules when soybean was inoculated with LPN-100. From these result we can suppose that the chemotaxis of Bradyrhizobium plays inportant the role of forming the nodule (host recognition) in the soybean-B. japonicum symbiosis.

• Journal of Plant Biology
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• v.37 no.3
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• pp.387-394
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• 1994

Effects of environmental factors of light, temperature, nitrogen sources and water stress were analyzed quantitatively on the nodule formation and nitrogen fixation activity of autumn olive plant (Elaeagnu$ umbellala Thunb.) during the seedling growth. Seedlings showed the maximum nitrogenase activity of $72.5\;\mu\textrm{M}\;C_2H_4{\cdot}g\;fr\;wt\;nodule^{-1}{\cdot}h^{-1}$ in the early nodulation stage. The relative growth rate and T/R ratio changed from $1.60%{\cdot}d^{-1}$ and 1.12 in the earlier stage to $3.75%{\cdot}d^{-1}$ and 2.31 in the later stage, respectively. light conditions of 20-25, 1015 and 4-6% resulted in decreases of 41, 54 and 71% of the nitrogenase activity, respectively. Nodules incubated in 15, 20, 25 and $30^{\circ}C$ showed the activities of 5.4, 24.7, 51.6 and $58.5\;\mu\textrm{M}\;C_2H_4{\cdot}g\;fr\;wt\;nodule^{-1}{\cdot}h^{-1}$ respectively. Pretreatment with low temperature ($15^{\circ}C$) followed incubation at $30^{\circ}C$ attained higher nitrogenase activity ($66.5\;\mu\textrm{M}\;C_2H_4{\cdot}g\;fr\;wt\;nodule^{-1}{\cdot}h^{-1}$) than that with higher temperature ($35^{\circ}C$). The oxygen pressure above 16 kPa is necessary for saturation of the nodule activity, but the activity was inhibited severely by physical impact such as the exision or isolation of nodules from the root. The relative activities of early nodules grown in pH 5.5, 6.5 and 8.0 were 89, 100 and 40% and those grown in 1 and 3 mM of $NO_3\;and\;NH_4$ were 6, 1 and 68, 50%, respectively. Watering levels of 20, 50 and 100 mL during the seedling growth resulted in 35, 120 and 8 mg of nodule formation and 33.6, 58.4 and $8.4\;\mu\textrm{M}\;C_2H_4{\cdot}g\;fr\;wt\;nodule^{-1}{\cdot}h^{-1}$ of the nitrogenase activity, respectively. Water stress with 86% decrease of soil water content caused temporary wilting point of leaf and a complete disappearance of nitrogenase activity of nodules, though the water content and transpiration rate in plant were reduced to 90 and 53%, respectively.tively.

• Environmental Engineering Research
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• v.20 no.1
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• pp.41-50
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• 2015

The cyanobacterium Anabaena doliolum Ind1 isolated from a coal mining site was tested for removal of cadmium at optimum pH 7.0 and temperature $25^{\circ}C$. The organism recorded high percentage of metal removal (92-69%) within seven days of exposure to 0.5-2.0 ppm cadmium. Biosorption onto the cell surface was the primary mode of metal removal. Fourier transform infrared spectroscopy (FTIR) established hydroxyl, amides, carboxyl, sulphate and carbonyl groups to be the major functional groups on the cell surface involved in cadmium binding. Cellular ultrastructure and a range of vital physiological processes (i.e., photosynthetic pigments, respiration, photosynthesis, heterocyst frequency and nitrogenase activity) remained unaffected upon 0.5 ppm treatment; higher concentrations of cadmium exerted visible adverse effects. Amongst the five photosynthetic pigments tested, phycocyanin was the most targeted pigment (inhibition was 15-89%). Both respiration and photosynthetic activities were inhibited by cadmium with more severe effect seen on respiration. 2.0 ppm cadmium exposure also had drastic negative effect on nitrogenase activity (87% decreased).

• Mycobiology
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• v.32 no.4
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• pp.173-178
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• 2004

Fifteen Rhizobium spp. from nodules of 6 common pulses collected from 6 districts of Assam were studied for their infectivity, intrinsic antibiotic resistance, nitrogenase activity and effect of dual inoculation with two native Arbuscular Mycorrhizal Fungi viz. Glomus mosseae(GM) and Gigaspora gilmarie(GG). Out of the 15 isolates 9 were found nodulation positive and 6 of them(AR1, BR8, BR12, AR10, UR10 & GR21) were subjected to intrinsic antibiotic sensitivity test of which AR1 showed resistance against all the 9 test antibiotics. Isolates AR1 and GR21 showed the highest(4.25 mole, $gm^{-1}hour^{-1}$) and the lowest(1.05 mole, $gm^{-1}hour^{-1}$) nitrogenase activity respectively. In Most Probable Number count, the maximum Rhizobium population $5.8{\times}10^5$, was found in both Blackgram and Greengram variety of pulses. The maximum dry weight of nodules(3.14 g), dry weight of shoot(10.08 g), nitrogen content(7.68 mg, $plant^{-1}$), chlorophyll content(1.89 mg, $g^{-1}$), phosphorus content of shoot(6.17 mg, $g^{-1}$) and yield(535.67 kg, $Ha^{-1}$) were found when AR1 dually inoculated with GM in Blackgram.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.151-157
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• 1990

The maximum nitrogen fixation activity of the associative, microaerobic and acid tolerant bacteria, Azospirillum amazonense Kp1 was obtained with 0.2Kpa of $O_{2}$ and showed a reversible inhibition by the higher concentrations. Ammonium treatment caused a gradual inhibition of the activity up to 350mM. The nitrogenase systems were purified by gradient chromatography on DEAE-52 cellulose, heat treatment and preparative PAGE. The MoFe protein showed molecular weight of 210,000 including two nonidentical subunits with apparent molecular weights of 55,000 and 50,000 and an isoelectricpoint of 5.2 and contained 2, 24 and 28 atoms of Mo, Fe and acid labile S per molecule. The Fe protein revealed molecular weight of 66,000 including two types of subunits with molecular weights of 35,000 and 31,000 and an isoelectric point of 4.6, and contained 4 atoms of Fe and 6 atoms of S per molecule. The maximum specific nitrogenase activity attained 2,200 and 1,700nM $C_2H_4mg^{-1} min^{-1}$, respectively for MoFe and Fe proteins at pH7 and $35^{\circ}C$. The activity was lost after 10 and 30 days under the cold room ($4^{\circ}C$) condition for Fe and MoFe proteins, respectively.

• Environmental Engineering Research
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• v.12 no.4
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• pp.136-147
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• 2007

In an effort to identify possible microbes for seeking bioagents for remediation of herbicide-contaminated soils, seven species of phototrophic nonsulfur bacteria (Rhodobacter capsulatus and sphaeroides, Rhodospirillum rubrum, Rhodopseudomonas acidophila, blastica and viridis, Rhodomicrobium vannielii) were grown in the presence of the herbicide, butachlor, and bacterial growth rates and nitrogen fixation were measured with different carbon sources. Under general conditions, all species showed 17-53% reductions in growth rate following butachlor treatment. Under nitrogen-fixing conditions, Rb. capsulatus and Rs. rubrum showed 1-4% increases in the growth rates and 2-10% increases in nitrogen-fixing abilities, while the other 5 species showed decreases of 17-47% and 17-85%, respectively. The finding that Rp. acidophila, Rp. blastica, Rp. viridis and Rm. vannielii showed stronger inhibitions of nitrogenase activity seems to indicate that species in genera Rhodobacter and Rhodospirillum are less influenced by butachlor than those in Rhodopseudomonas and Rhodomicrobium in terms of nitrogen-fixing ability. Overall, nitrogenase activity was closely correlated with both growth rate and glutamine synthetase activity (representing nitrogen metabolism). When the carbon sources were compared, pyruvate (three carbons) was best for all species in terms of growth rate and nitrogen fixation, with malate (four carbons) showing intermediate values and ribose(five carbons) showing the lowest; these trends did not change in response to butachlor treatment. We verified that each of the 7 species had a plasmid ($12.2{\sim}23.5\;Kb$). We found that all 7 species could use butachlor as a sole carbon source and 3 species were controlled by plasmid-born genes, but it is doubtful whether plasmid-born genes were responsible to nitrogen fixation.

• The Korean Journal of Ecology
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• v.18 no.1
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• pp.43-54
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• 1995

Effects of environmental factors of N, P, pH, moisture, temperature and oxygen on growth and nitrogen fixation activity of kummerowia striate (Thunb.) Schindler seedling, bearing symbiotic root nodules, were quantitatively analyzed during the growing period. The specific nitrogenase activity (ARA) of nodules showed the maximum value of 187 μmol C₂H₄g fr wt-1 h-1 6 weeks after seeds were germinated. The total nitrogenase activities per plant attained as 1.56, 0.85, 0.09 and 4.0, 1.11, 0.04 μmol C₂H₄hr-1, respectively for the treatments of 1, 3 and 5 mM NO₃￣and NH₄＋ on the 60th day. While the plant grown in N-free media for 20 days after treatments of 5 mM NH₄+for 40 days resulted in 30 mg fr wt of nodule formation and exhibited the relative activities of 152％ and 162％ for total and specific ARA in comparison with those of control plant grown with N-free for 60 days. Total biomass and ARA was by 70％ and 86％ lower in N and P deficiency, respectively. The N and P deficient plot showed 70％ and 86％ decreases of total biomass and ARA in comparison with those of control. The plant grown with N-free for 20 days after pretreatment with N and P free media for 40 days showed the relative values of 77％, 118％ and 150％, respectively for nodule biomass, total and specific ARA in comparison with those of control. The treatment with acid or alkali gradients resulted in significant decreases of nodule biomass and ARA. The optimum temperature and pO₂for ARA were 30°C and 40 kPa, respectively. Two peaks of diurnal variation appeared at 11:00 and 23:00 o'clocks by the continuous light condition. The plants with water stress by temporary wilting point rsulted in 95~97％ inhibition for nodule respiration, transpiration and specific ARA. Transpiration and ARA ware recovered to 88％ and 38％ of those of water unstressed plants, respectively, 6 hours after the plants were rewatered from water stressed condition.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.4
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• pp.308-315
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• 2003

A field experiment was conducted to investigate the performance of three legume species in a zero-tillage, non-fertilized rice field in a temperate zone. Before the experiment for 5 years, from 1995 to 1999, plant growth patterns of three legume species grown as over-wintering (October-May) cover crops on a paddy field were maintained to study N balance and $\textrm{N}_2$ fixation. Decrease in plant density accelerated from after winter to flowering from 1,090, 320, and 5 to 732, 232, and 6 plants $\textrm{m}^{-2}$ in Chinese milk vetch (CMV), white clover (WC), and hairy vetch (HV), respectively. Total dry weights of plants above-ground level were 0.05, 0.11, and 2.43 g $\textrm{plant}^{-1}$. in WC, CMV and HV respectively but steeply increased at ripening up to 0.77, 2.33, and 26 g $\textrm{plant}^{-1}$. The root dry weight of HV and CMV rapidly increased while in WC, root dry weight increased slightly towards flowering. The highest nodule numbers were recorded in CMV to April thereafter WC produced the highest. Nodule size was distributed within 7mm in CMV but it was larger in HV varying from 1 to 10mm. Shoot N (g $\textrm{m}^{-2}$) greatly increased from over-wintering to flowering in CMV, HV and WC and it ranged from 1.66, 0.5 and 1.92 to 12.6, 3.1 and 13.02 g $\textrm{m}^{-2}$, respectively. After wintering, the initial shoot N content (%) was more in CMV. Root N content (%) was constant or slightly decreased in HV and WC. Soil total N in the control plot (clean fallow) was the highest on Mar. 2 then decreased rapidly to flowering. Soil N content was constant in HV plots whereas it was low in WC plots for the entire growth period except just after winter. Maximum nitrogenase activities were 9, 37.8, and 131 mol $\textrm{C}_2\textrm{H}_4$ $\textrm{plant}^{-1}$ $\textrm{hour}^{-1}$. in CMV, HV, and WC, respectively. Nitrogenase activity showed a direct correlation with nodule number, size and fresh weight. As a cover crop preceding a rice crop, CMV is more suited to colder regions due to its earlier ripening characteristics. Hairy vetch and WC are recommended for regions with a mild winter and a long summer owing to their late ripening and great N fixation activity.

• Journal of Environmental Science International
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• v.21 no.9
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• pp.1139-1147
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• 2012

To find out the growth conditions for the maximum activity of nitrogenase which catalyzes nitrogen fixation in Rhodobacter sphaeroides, the promoter activities of nifA and nifH were analyzed and the results indicated that expression of both nifA and nifH was increased in response to deprivation of both O2 concentration and nitrogen source. The nifA mutant was constructed by deleting the gene to investigate the effect of NifA, the transcriptional regulator, on the nifH and nifA expression in R. sphaeroides. Analysis of expression of nif genes using the nifA::lacZ and nifH::lacZ fusions in the nifA mutant revealed that NifA acts as a positive activator for nifH and an autoregulator in its own expression. The promoter activities of nifA and nifH in the prrA mutant grown under anaerobic and ${NH_4}^+$-free conditions were derepressed, comparing with those of the wild-type grown under the same conditions, indicating that the prrA product acts as a positive regulator in expression of nifA and nifH.

• Korean Journal of Microbiology
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• v.29 no.6
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• pp.408-415
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• 1991

TOL plsmid size of Flavobacterium odoratum SUB53 was estimated as 83 Md and the optimum concentration of m-toluate degradation by TOL plasmid was 5 mM. $H_{2}$ production by Rhodopseudomonas sphaeroides KCTC1425 was largely dependent on nitrogenase activity and showed the highest at 30 mM malate with 7 mM glutamate as nitrogen source. Nitrogenase activities were inhibited by 0.3 mM $NH_{4}^{+}$ions, to be appeared the decrease of $H_{2}$ production. Conjugation of TOL plasmids from F. odoratum SUB53 and Pseudomonas putida mt-2 to R. sphaeroides showed the optimum at the exponential stage of recipient cells in presence of helper plasmid pRK2013. According to the investigation of catechol-1,2-oxygenase (C-1, 2-O) and catechol-2,3-oxygenase (C-2,3-O) activities of R. sphaeroides C1 (TOL SUB53) and C2 (TOL mt-2), the gene for C-2,3-O is located on TOL plasmid and gene for C-1, 2-O on the chromosome of R. sphaeroides. m-Toluate was biodegraded by TOL plasmid in R. sphaeroides C1 and C2, presumably to be produced $H_{2}$ gas from the secondary metabolites of m-toluate.e.