• 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.

• 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 The Korean Society of Grassland and Forage Science
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• v.6 no.3
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• pp.151-156
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• 1986

There are differences in recommendations in the USA as to whether nitrogen fertilizer should be applied when establishing alfalfa (Medicago sativa L). The reason for not applying nitrogen is because some researchers found the addition of nitrogen reduced nodulation of alfalfa plants. A replicated experiment was conducted under controlled environmental conditions at the University of Nevada-Reno, Reno, Nevada, USA, to determine the effects of nitrogen application on seedling growth and nodulation of alfalfa when grown in a cool environment. A sterile sand was used in the growing media to which a complete nutrient solution minus nitrogen was applied volumetrically to each pot daily. Half of the pots received $NH_4NO_3$, at the rate of 11.2 kg/ha, at seeding and two and four weeks after planting, giving a total nitrogen application rate of 33.6 kg/ha. Rhizobia inoculant (R-12) consisted of a mixture of strains 171-15a. 1682c and 80 PI 265 of Rhizobium meliloti. Inoculant was applied to the seeds prior to planting and to the sand media at two and four weeks after seeding. Twenty seeds were planted in pots 14.0 cm in diameter and 11.5 cm deep. Seedlings were thinned after emergence to ten plants per pot. They were grown in a controlled environment chamber with a 16-hour light period. Soil temperatures at 6 cm depth ranged from $5.7^{\circ}C\;to\;21.5^{\circ}C$ and had a daily mean of $16.2^{\circ}C$ Plants were harvested at weekly intervals for seven weeks at which time root, shoot and total length, dry weight, volume and number of nodules per plant were determined. Root, shoot and total length were not affected by nitrogen fertilizer. However, application of nitrogen increased the size of the seedlings as determined by dry weight and volume when compared to plants which were not fertilized. This indicates that rhizobia did not fix enough atmospheric nitrogen to promote good growth. Nitrogen application resulted in significantly more nodules per plant. The effect of nitrogen fertilizer became more apparent as the plant became older. Results of this experiment show there are benefits from applying nitrogen at a low rate when establishing alfalfa under a cool environment.

• Korean Journal of Environment and Ecology
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• v.20 no.4
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• pp.425-435
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• 2006

The objectives of this research were to investigate the drought resistance as well as the growth of Wisteria floribunda seedlings with the soil water conditions. The seedlings for the research were grown in pots with strict water content control on a frame located outdoors. During the experiments, the soil water contents were adjusted to 5, 10, 20, 30, 40, 50, 60, and 70%. In addition, the effects of Rhizobium inoculation on the growth of seedlings were investigated. The experimental results are summarized as follows: 1. The seedlings in the pots with 5% soil water content withered to death due to the water stress. Withering or any distinct growth was not observed from the seedlings in the 10% soil water content. It can be inferred from these results that about 5% of soil water content is the wilting point of W. floribunda seedlings and about 10% is the critical soil water content of its growth in this experiment soils. Therefore, it seems that W. floribunda possesses a good drought resistance. 2. From the analyses of the main growth parameters such as stem elongation, diameter growth, leaf area growth and total dry weight, it was found that the seedling growth can be improved with increasing soil water contents. The relation between each growth parameter(Y) and the soil water contents(W) was well described by a quadratic equation, $Y=a+bW+cW^2$. 3. In soil water contents higher than 20%, the seedling growth(Y) was remarkable along with-its extended growing period, and related to the growing period(D) by a quadratic equation, $Y=a+bD+cD^2$. 4. The artificial inoculation of Rhizobiun promoted the growth of Wisteria floribunda seedlings. 5. Rhizobium was found to be more readily inoculated and to form more root nodules compared to seedlings grown in higher soil water contents.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.258-263
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• 1994

Genomic Southern hybridization of Frankia EuIKl strain, a nitrogen fixing symbiont of Elaeagnus umbellate root nodules, with nifH,D of K. pneumoniae as a probe, showed that 3.2 Kb and 5.5 Kb of BamHI fragments and 15 Kb PstI fragment were strongly hybridized with the probe, indicating nifH,D are located on these fragments. Using the same probe, one clone(pEuNIF) was isolated from the genomic library constructed into pWE15 cosmid vector by colony hybridization. The 3.2 Kb and 5.5 Kb BamHI fragments of this clone were hybridized with the same probe and this result corresponds to the genomic Southern hybridization data. However, using nifH of Frankia FaCl strain as a probe, only the 3.2 Kb BamHI fragment showed hybridization signal. Amino acid sequence deduced from nucleotide sequence of 3' terminus of the 3.2 Kb and 5' terminus of the 5.5 Kb fragments showed that the former was highly homologous with that of ArI3 nifD from 182nd to 240th amino acids, while the latter was from 241st to 282nd amino acids. These results show that nifH and partial nifD sequences are located on the 3.2 Kb fragment and residual sequences of nifH on the 5.5 Kb fragment which is contiguous to the 3.2 Kb fragment.

• Journal of The Korean Society of Grassland and Forage Science
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• v.15 no.4
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• pp.245-252
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• 1995

To select a carrier as Rhizobiwn inoculants for alfalfa, severeal Rhizobium inoculants were produced by adding or not sub-materials($CaCO_3$ and sucrose) to main materials(organic or inorganic materials). The root nodule bacteria, Rhizobium meliloti 1061 distributed from KAIST, and Vemal alfalfa was used in this experiment. The Rhizobium populations and inoculation effects of the Rhizobium inoculants produced in several materials were scrutinized at laboratory and greenhouse in Livestock Experiment Station, RDA from 1993 to 1994. Moisture contents of the caniers were varied from 32 to 50% on dry weight basis according to material characteristics and the pH ranges of these were varied from pH 4.56 to 10.06 according to raw material characteristics and preparations. Initial Rhizobium numbers of the carriers were higher in organic material-inoculants than in inorganic, and among the inoculants, the inoculant made of Bentonite+Vermiculite(l:3 w/w) was excellent because of high rhizobium population($7.8~8.3\times10^8/g$ inoculant) and high rhizobium reappearance of inoculant in severed different production time. The root nodules of the alfalfa inoculated with different inoculants were fast formed in the fermented sawdust with cattle dung (FSC) inoculant, and bentonite(B)+vermiculite(V) than others. Plant length of alfalfa was differentiated on 15 days after inoculation but was not nearly different between higher inoculants than rhizobium number $10^7/g$ inoculant. Total dry matter of alfalfa was yielded by 20.65, 20.34mg per pot in FSC + sucrose 0.5% and B + V + sucrose 1% inoculants respectively that were higher inoculation effect by 17 times compared with non-inoculation, 1.2 mg per pot.

• Korean Journal of Microbiology
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• v.30 no.4
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• pp.246-251
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• 1992

Molccular cloning of nod genes from Bradvrhizobium sp. SNU001, a nitrogen-fixing symbiont isolated from thc root nodules of soybean (Clycine trim) . was carried out. nod genes were found to be located on thc genome of the symbiont by gcnomic hybridization with 4.5 kb EcoRI/HndIII fragment (nod DABC) of Rhizohium meliloti as probe. Genomic library of this symbiont was constructed using h phage EMBL3-BanlHI vector. from which five nod positive clones were sclectcd by primary and secondary screening methods. The partial restriction map of inserted genomic DNA of h CNS-l(c1one 2) was constructed. and 3.9 kh Bun7HI fragment. which showed strong hybridization signal to the probe, was subcloned into pBS KS(+) plasmid vector. Partial restriction inap ot' a selected subclone (pBjCNS-I) was constructed and nod DABC was found to be located on the 1.8 kb KpnI/Sacl fragment of this subclone.

• Restorative Dentistry and Endodontics
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• v.22 no.1
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• pp.305-324
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• 1997

Numerous materials such as amalgam, IRM, SuperEBA, dessicated ZOE, and Ketac-Silver have been used as a root-end filling material or to repair furcation perforations. But so far no material has been found to satisfy all of the requirements of an ideal restorative material. Recently, mineral trioxide aggregate (MTA) has been suggested for use as a root end filling material and for the repair of furcation perforations. The purpose of this study was to compare the effect of MTA on the proliferation of MC3T3/E1 osteoblastic cell, formation of bone nodule, alkaline phosphatase activity, and finally the tissue reaction of bone with those of amalgam, IRM, SuperEBA, dessicated ZOE, and Ketac-Silver. The following conclusions were drawn within the limits of the experimental results : 1. MTA showed a excellent proliferation of osteoblastic cell and Ketac-Silver showed moderate proliferation of osteoblastic cell. The rest of test materials showed no proliferation of osteoblastic cell. 2. Many of definite bone nodules were found in the MTA group. In contrast, Ketac-Silver group showed no definite bone formation but only showed mild sign of bone formation. 3. Alkaline phosphatase activity of Ketac-Silver and MTA showed similar results. But both of them showed higher activity than that of other materials (p<0.005). 4. The tissue reaction to implanted MTA in the calbarium of mouse was milder than that observed with other materials. The tissue reaction of dessicated ZOE showed the worst results among the test materials.

• 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.39 no.4
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• pp.389-396
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• 1994

Poor establishment of white dover (Trifolium repens L.) into grass-dominant pastures has been limited its availability. The experiment was done to clarify the effects of timing of initial defoliation, defoliation frequency on the regrowth and nodule formation of the clover cultivars during 28-day regrowing period. Individual plants of cv. Regal, Louisiana S-1 (La. S-1), Grasslands Huia (Huia) and Aberystwyth S184 (S184) were grown in containers until grown to unifoliolate, 1, 2, 4, or 8 trifoliolate stage, and then clipped to 1cm in height every 7 or 28 day for 28 days. To measure the effects, plants were sampled immediately after final harvest, and 1, 3, 7, 14 and 28 days after the harvest. Shoot, root dry weight and biomass were reduced with earlier, more frequent defoliation or shorter regrowing period. In frequent defoliation shoot dry weight and biomass were increased with delayed initial defoliation while in less frequent defoliation steeply done when initial defoliation was delayed to 4 trifoliolate stage. Shoot /Root ratio inclined with more frequent defoliation or lengthened regrowing period, and was greater in initial defoliation of unifoliolate to 2 trifoliolate than the others. Although nodules no. per plant declined with earlier or more frequent defoliation, the effect disappeared to some extent after 14-day regrowth. In comparison with the others, Regal had the highest shoot. dry weight and biomass to 2 trifoliolate stage while S184 did the most nodules regardless of defoliation timing. On 7-day after last defoliation nodule formation of Regal, Huia and S184 but on 28-day after last defoliation that of La. S-1, Huia and S184 was positively correlated to shoot and root dry weights upto 2 trifoliolate stage. On the former day, however, that was negatively correlated to Shoot /Root ratio upto 1 trifoliolate stage although on the latter day it was not, meaning that in addition to more frequent defoliation earlier defoliation was harmful in nodule formation of white clover.