• Applied Biological Chemistry
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• v.41 no.5
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• pp.355-362
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• 1998

The minor form of phosphofructokinase (EC 2.7.1.11; PFK), which was suggested to be of plastid origin from the host fraction of chickpea nodules, was isolated as a small protein with apparent molecular mass near 220 kDa and purified to a high degree. SDS-PAGE and western blot indicated that the enzyme was made up of a homotetrameric structure (55 kDa). The enzyme had sharp pH profiles with maximal activities at pH 8 and displayed Michaelis-Menten kinetics with respect to Fru-6-P and nucleoside triphosphate substrate at the pH optimum (pH 8) and at pH 7. MgATP was the most effective phosphoryl donor. Phosphoenolpyruvate was a potent inhibitor of minor PFK activity, and the enzyme was also strongly inhibited by 3-phosphoglycerate, 2-phosphoglycerate, and to a lesser extent, PPi. Minor PFK was weakly activated by KCl, NaCl and Pi, and was inhibitory at high concentration of KCl and Pi.

• Journal of The Korean Society of Grassland and Forage Science
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• v.23 no.2
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• pp.81-90
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• 2003

This pot experiment was conducted to find out the effects of boron application(B$_{0}$; 0.0, B$_1$; 0.2, $B_2$;2.0, $B_3$;6.0, $B_3$;6.0, $B_4$;15.0me B/pot) on the forage performance in the pure and mixed swards of orchardgrass and white clover. This 1st part was concerned with the changes in the growth, flowers and flower buds, and roots/nodules of forages. The results obtained are summarized as follows: 1. At the $B_3$ and $B_4$ treatments, the B toxicity was more seriously in white clover than in orchardgrass in the first half of cutting orders, and reduced in the latter half. In white clover, it was more worsened in a mixture than in a pure culture. It was somewhat reduced at the best growth stage of each forage. 2. In orchardgrass, the B toxicity($B_3$,$ B_4$) showed the chlorosis on leaf tips, shallow leaf, little tillers, and weak stems. Whereas it showed the chlorosis/necrosis on old leaf edge, little and weak stolons in white clover. 3. Comparing with the B deficiency($B_{0}$ , $B_1$) and toxicity($B_3$, $B_4$), the optimum B application($B_2$) influenced markedly good growth of shoot, root, nodule, and flower (flower number, blooming period, early full flower) in white clover. 4. Comparing with orchardgrass, white clover was greatly influenced by the boron application. However, this responses of white clover to boron were reduced in a grass-clover mixture with additional fertilization. It was recognized that the good forage performance in a grass-clover mixture could be regulated by the adequate applications of boron and additional fertilizers.s.

• Applied Biological Chemistry
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• v.35 no.6
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• pp.449-456
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• 1992

Expermient were conducted to determine the effect of phosphorus stress on bacteroid content and energy status of soybean (Glycine max [L.] Merr.) nodules. Plants inoculated with Bradyrhizobium japonicum MN 110 were grown with P-stressed (0.05 mM-P) and control (1 mM-P) treatment in the greenhouse. Phosphorus stress decreased nodule mass per plant and nodule mass to whole plant mass ratio. Phosphorus concentration in leaf, stem and root tissues were reduced by 75% but in nodule tissue was reduced only by 40% under phosphorus stress during 3 week experimental period. The bacteroid content per unit nodule mass and the distribution of total nitrogen and total phosphorus among the bacteroid and plant cell fractions of nodule were not affected significantly by phosphorus stress. Regardless of phosphorus treatment, 22% of the nitrogen and 27% of the phosphorus in whole nodules were associated with the bacteroid fraction. The ATP and total adenylate concentrations in and energy charge of whole nodule were decreased 77%, 46% and 37%, respectively, by phosphorus stress. The ATP concentration in and energy charge of the host plant cell fraction of nodules were decreased 86% and 59%, respectively, but these parametres in bacteroid in nodules were not affected by phosphorus stress. These results indicated that nodule is a strong phosphorus sink and that nodule growth and development are regulated to maintain a high phosphorus and energy content in bacteroid even when the host plant is subjected to phosphorus deficiency.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.51-57
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• 1970

On the basis of the specific interrelationship between the species or variety of leguminous crops and the species or strain of nodule bacteria, Rhizobia, the rhizobial species and strain must be effectively chosen for the successful inoculation. The present paper describes on some results of the isolation and taxonomic study on the native rhizobial strains isolated from the nodules of five species of leguminous crops such as numerous varieties of soy bean, lespedeza, birdfoot trefoil, ladino and red clovers. The isolated strains of soy bean nodule bacterium, Rhizobium japonicum were grouped through the inoculation test on variety Changdanbaikmock into the effective, noneffective and toxic strain for the nodule formation. In the study of the effect of some inorganic and organic nitrogenous compounds on the growth of Rhizobium japonicum strain Ac 20, a promotive response was showed by asparagine, and glutamine, but hydroxylamine, nitrite, hydrazine and azide was inhibitory at the concentration of $10^{-2}M/l$ in mannitol-yeast extract basal medium. In the physiological characteristics each strain showed somewhat different activities of the indole-3-actic acid formation and hydrogenase and discussed with these characters in relation to nodule forming ability.

• Current Research on Agriculture and Life Sciences
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• v.15
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• pp.25-32
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• 1997

This study was carried out to research the biological characteristics among rhizobia and soybean seed and root lectins, and to obtain a basic imformation of host specificity in biological nitrogen symbiosis system. The results obtained were as follows: Purified seed lectin from soybean varieties of paldal, backwoon and hwangkeum formed immunoprecipitin lines with standard soybean seed lectin and the root lectins from soybean seedlings immunoelectrophoretically. Soybean seed and root lectins interacted with Rhizobium japonicum and Bradyrhizobium japonicum, but didn't interacted with Rhizobium. viceae, whereas pea lectin conjugated with R. viceae, but didn't bind with R. japonicum and B. japonicum. Lipopolysaccharides of B. japonicum and R. viceae were fractionated into LPS I and LPS II on the sephadox G-50. Lipopolysaccharides from B. japonicum showed rhe binding acitivity both with soybean seed lectin and root lectin, but those from R. viceae didn't show it with soybean seed and root lectins.

• 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.

• Korean Journal of Soil Science and Fertilizer
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• v.15 no.4
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• pp.277-283
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• 1982

A series of experiments were conducted on procedures of $N_2$ fixation measurements with soybean (Glycine max. L.) root nodules by acetylene reducing assay and estimation of amounts of $N_2$ fixed by soybean root nodules grown in the pot. The results are summarized as follows; 1. In case of nitrogen fixing activities were determined by means of acetylene reducing method, more appropriates results were obtained with direct determination of root nodules which was adhering from root under 2 hours after incubation, while more fluctuated data was obtained with root nodules determined under separated conditions. 2. The weight of soybean nodules was linearly increased from 17 days to 55 days after planting, whereas, the amounts of $N_2$ fixation from soybean nodules were Peaked at 38 days after planting, and declined thereafter. 3. When calculated upon the basis of the amounts of $N_2$ fixed as measured by acetylene reducing activities in single soybean plant, it was found that the average amount of symbiotic $N_2$ fixation falls in the range of 80 kg N per ha.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.1
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• pp.16-21
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• 1992

Increase in nodulation and nitrogen fixation was achieved partly through the isolation of supernodulating soybean mutant plants. This experiment was conducted to compare nodulation, nitrogen fixation, and early growth characters of wild type 'Bragg' with those of its supernodulating soybean mutant, 'nts 382' and 'nts 246'. At 31 days after planting, nodule dry weight of nts mutants was 2.5 to 3.7 times greater than that of Bragg. Higher nodulation of nts mutants showed the reduced top growth, indicating that photosynthates might be translocated and used for nodule growth attached to the root system. Total acetylene reduction activity was higher in nts mutants than Bragg, whereas specific acetylene reduction activity of nts mutant was the half of that of Bragg. Mixture of nts mutants and Bragg did not affect nodulation characters each other, suggesting that factors affecting supernodulating characters exist inside rather than outside the root system.

• 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.

• Journal of The Korean Society of Grassland and Forage Science
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• v.24 no.2
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• pp.105-114
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• 2004

This pot experiment was. conducted to investigate the effects of systematic variation application of Fe, Mn, Cu, and Zn on forage performance of orchardgrass and white clover. The treatments of systematic variation were 0/100, 25/75, 50/50, 75/25, and 100/0％ in the Fe/Cu(trial-1), Mn/Zn(trial-2), and Fe + Cu/Mn + Zn(trial-3), respectively. The treatments of Fe/Mn/Cu/Zn(trial-4) were composed of 70％ in main element and 10％ in other 3 elements, respectively. 1. In general, the unbalanced applications of Fe and Mn resulted in the Mn and Fe deficiencies(chlorosis) on white clover, respectively, because of the antagonism between Fe and Mn. In white clover, the traits of growth, root/nodule, and flowering, which were influenced by the systematic variation of Fe, Mn, Cu, and Zn, were closely correlated to each other. In the Fe/Cu trial, the 0/100 and 25/75 induced a Fe-deficiency on white clover, and the 0/100 also showed poor root growth and flowering. In addition, the 50/50 and 75/25 showed an early flowering of white clover. 2. In the Mn/Zn trial, the 0/100 induced a severe Mn-deficiency(chlorosis) on white clover. The 25/75 and 50/50, however, diminished the chlorosis symptom. The 75/25 and 100/0 showed generally good root growth and flowering of white clover. 3. In the Fe + Cu/Mn + Zn trial, the 0/100 induced a Fe-deficiency, and the 100/0 induced a Mn-deficiency on white clover, which were correlated to the poor root growth and flowering. The 75/25 showed good root growth and flowering of white clover. 1be flowering of white clover tended to be more influenced by the Fe + Cu than by the Mn + Cu ratios. 4. In the Fe/Mn/Cu/Zn trial, the Fe and Mn deficiencies on white clover, which were influenced by the Mn and Fe treatments, also occurred. The Cu and Zn-deficiency symptoms, however, were not recognized. General differences have been showed in the numbers of flowers as following orders; Zn > Cu > Mn > Fe - 70% treatments.