• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.2
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• pp.198-206
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• 1981

A field research was conducted to evaluate the effects of inoculation of Rhizobium meliloti and lime application on the early growth of alfalfa, var. Luna, in an acid soil. Alfalfa seeds were inoculated by lime-peat pelleting method with $A_3$ strain isolated from alfalfa roots. Both inoculated and noninoculated seeds were sown in acid soil (pH 5.5) with three levels of lime application (1, 3, 6 ton/ha) on mid-April. Inoculated seeds produced many effective nodules in early stage and increased nitrogen and chlorophyll contents of leaf, consequently stimulating alfalfa growth. Pseudo nodules were found in noninoculated, while they were seldom formed in inoculated. Lime application increased soil pH, plant height, root length, and shoot and root weight. Liming effect on nodulation was significant at first cut, but no longer after then. Both inoculation and liming increased the forage yield of alfalfa. Inoculation and liming of 6 ton/ha together produced the highest yield. As compared to control plot of nitrogen application, the effect of seed inoculation only could be equal to about 120 kg/ha nitrogen application. The total nitrogen contents of forage were little affected by number or weight of nodules and amounts of lime application.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.3
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• pp.299-306
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• 2009

In nitrogen-limited conditions, rhizobia lead to formation of nitrogen-fixing nodules on the roots of leguminous plants. The process of nodulation is autoregulated by pre-existing nodules in the same root system. The altered profile of sap proteins by inoculation with B. japonicum may indicate presence of a signal responsible for autoregulation transferred through stem. The 20 kDa protein enhanced by innoculation significantly decreased in intensity from 2.5 to 7 days after inoculation (DAI). However 6 kDa protein did increase during such a transition period. Western blot analysis showed that both 20 kDa and 6 kDa were cross-reacted with the SKTI antiserum. This suggests that SKTI may be involved in soybean nodulation by specific induction and degradation in stem sap during early stage of nodulation. RNAi technique and Agrobacterium rhizogenes-mediated transformation were applied to investigate the function of SKTI in nodulation. We have found that the number of rhizobium-induced nodule was much less in SKTIi-silenced hairy roots than the non-silenced. Indeed the quantitative RT-PCR showed that the expression level of SKTI gene was reduced over 40% in the transgenic hairy roots compared to the non-transgenic. It appears that the observed early induction of SKTI and degradation into small peptide in a specific time manner may be involved in autoregulation of nodulation in soybean and the specific mechanism of such regulation remains to be investigated.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.32 no.3
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• pp.323-329
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• 1987

Five soybean varieties of two early maturing Karikei73 and SS 79168, and three late maturing Tohoku76, Baegunkong and Jangbaegkong were used and evaluated in the study. Of the varieties examined, Karikei73 was characterized by the delayed leaf senescence. The varieties were planted in the pots of 1/3500 a filled with volcanic ash soil at the experimental fields of the National Institute of Agrobiological Resources in Japan. Major agronomic characteristics including the activity of nitrogen fixation for root nodules during the grain filling period were measured. Measurements during the stages were followed by the stage of development descriptions for soybeans made by Fehr and Carviness (1977). The acetylene reducing activity (ARA) per dry weight of root nodule measured using acetylene reduction assays was the highest at R4-R4.5 with decreasing trends thereafter for the early matruing varieties, while it continuously increased up to R6 but decreased rapidly thereafter for the late maturing varieties. The dry weights of root nodules and all parts of the host plant at each stage checked were greater in the late maturing varieties being the same in ARA per pot.

• Current Research on Agriculture and Life Sciences
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• v.5
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• pp.40-51
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• 1987

This experiment was conducted to search for Rhizobia with good nitrogen fixation abilities and to investigate their physiological characteristics isolated from 12 soybean cultivars and the affinities of root nodule bacteria with soybeans. The results obtained were as follows; Based on colors, Rhizobia grown on YMA medium were divided into 3 groups, i. e., white, translucent and transparent, amounting to 60, 30 and 10%, respectively. In litmus milk reaction, the strains which produced alkali, acid serum, alkaline serum and acid reached to 51, 29, 9 and 11%, respectively. Strains, S022, and S096 were slow-growers and produced alkaili, while strains, S080, S-090, and S118 were fastgrowers and produced acid. The growth of root nodule bacteria on YMA medium was favorable between the initial pHs of 6.0~7.0. Glutamine, asparagine and al1antoin as nitrogen sources enhanced the growth of root nodule bacteria. All the strains tested formed nodules on the soybean roots, and the strains with good symbiotic nitrogen fixation abilities that had white color, small colony, nitrate reduction abilities and no nitrite reduction abilities showed comparatively high nitrogen fixing activities. Some strains varied in nitrogen fixing activities according to soybean cultivars, and a few strains fanned ineffective nodules which showed no nitrogen fixing activity.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.450-455
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• 1992

Five strains of the fast-growing endosymbionts were isolated from the nodules of Lotus eorniculatus var. japonicus inhabited in Taejeon. From morphological and physiOlogical characteristics and nodulation test, the isolated strains were identified as Rhizobium loti. Compared to the control plant, both Lotus cornieulatus var. japonieus and Lotus corniculatus seedlings inoculated with the isolated strains. grew normally due to effective root nodule. The reisolated endosymbiont from the induced root nodule was confirmed identical to those of the first isolates by investigating antibiotic resistance and morphological characteristics. Three strains among the isolates. R. loti TUS I. TUS5 and TUS6 produced a ca1cof1uorbinding exopolysaccharide.

• The Korean Journal of Ecology
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• v.21 no.3
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• pp.179-186
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• 1998

Effects of lead(Pb) and calcium(Ca) on growth responses, accumulation of heavy metals and nitrogen fixation activities of Melilotus suaveolens seedlings were quantitatively analyzed during growing period. Pb contents of the root treated with 30, 100 ppm Pb and 100 ppm Pb added 100 ppm Ca were 54.1, 90.9 and 26.1 folds higher than that of the control, respectively, at pH 4.2 in 28th days, and heavy metal content of plant increased with increasing of pH and Pb concentration. The melilot plant was classified as a Pb accumulator by higher accumulation of Pb in shoot than that of root. Pb treatments resulted in inhibiton of height and chlorophyll contet, and Ca treatment increased height and chlorophyll content insignificantly at pH 4.2 in 28 days. The plant biomass reduced 49, 60 and 54% at pH 4.2 and 47,53 and 50% at pH 6.5, respectively, by 30, 100 ppm Pb and 100 ppm Pb added 100 ppm Ca treatment. Specific nitrogen fixation of nodules reduced 68.4% and 46.6% by 100 ppm Pb treatment and 3.7% and 24.9% by 100 ppm Pb added 100 ppm Ca at pH 4.2 and pH 6.5, respectively, so Ca inhibited significantly Pb activity and toxicity in acdic pH. Nodule formation were reduced to 33, 33 and 50% at pH 4.2 and 50, 33 and 38% at pH 6.5 by 30, 100 ppm Pb and 100 ppm Pb added 100 ppm Ca, respectively.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.4
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• pp.265-267
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• 2005

Study on effectiveness of Bradyrhizobium japonicum on soybean [Glycine max (L.) Merr.], local cultivar 'Sathiya' were carried out in the garden soil of Trib-huvan University. Different parameters like nodulation, chlorophyll content in fresh leaves and growth of plant in inoculated and uninoculated plant was studied. Pot experiment was conducted in the green house to evaluate the effectiveness of B. japonicum on soybean. It was observed that B. japonicum inoculation increased the number of nodules, shoot length of plant and total chlorophyll content in fresh leaves of soybean plant. However, root length was decreased in all inoculated plants.

• Korean Journal of Microbiology
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• v.35 no.2
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• pp.169-172
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• 1999

Small-seeded legumes can be cultured enclosed in slant agar tubes if dlese plants are to he used for authenicaiing rhizobia or for enumerating 1-hizobia by the plant-inlection technique. An improved method has been developed with substiluting agar slant lor Korean paper(Har7ji). This method was panicularly useful for legumes with rigid radicle such as Cn.xsin 1mnmame. With this method Bmr!,~li1rzobin,17 sp. stram CN9135 on C nonmne induced root nodules biginning at day 7 of the nodulation period in 6% of the l ~ l a ~ l s , and all of ihe plants nodulated 14 days after inoculation by strain CN9135.

• Journal of The Korean Society of Grassland and Forage Science
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• v.16 no.2
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• pp.113-120
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• 1996

The experiment was carried out to investigate the effects of phosphate fertilizer level on growth, crude protein content and nitrogen fixation ability of alfalfa(Medicugo sariva L.) at sowing year. The experimental field was conducted using a randomized block design with three replications according to the phosphate application level (200 and 400kfia) in farm of Kongju National University. Research samples were collected at intervals of 7 days from October 7 to December 9 and nitrogen fixation activity of nodule was measured by using the acetylene reduction method (Hardy et al. 1968). The results obtained are summarized as follows : 1. Number of branches, root length and dry matter weight at phosphate fertilizer level of 400kg in sowing year were more increase than those of 200kfia. The number of nodules was not significantly difference between phosphate fertilizer level, because of low temperature to be enough to nodulation of alfalfa. 2. The crude protein content of leaf, stem and root were not significantly affected by the phosphate fertilizer level. but that of leaf was increased at 400kgha on 72 days after sowing. The crude protein content of each part according to growing stages was increased at 44 days and 72 days after sowing. 3. The dry weight of root was not significantly difference between phosphate fertilizer level from 30 days to 51 days after sowing, but that was increased at phosphate fertilizer level of 400kgha from 51 days afier sowing. 4. The amount of ethylene in the samples taken 2 hours and calculated amount of nitrogen fixation were increased at phosphate fertilizer level of 400kfia from 30 days to 65 days after sowing. but there was vice versa from 58 days after sowing.

• Molecules and Cells
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• v.24 no.2
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• pp.185-193
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• 2007

Symbiotic nitrogen fixation with nitrogen-fixing bacteria in the root nodules is a distinctly beneficial metabolic process in legume plants. Legumes control the nodule number and nodulation zone through a systemic negative regulatory system between shoot and root. Mutation in the soybean NTS gene encoding GmNARK, a CLAVATA1-like serine/threonine receptor-like kinase, causes excessive nodule development called hypernodulation. To examine the effect of nts mutation on the gene expression profile in the leaves, suppression subtractive hybridization was performed with the trifoliate leaves of nts mutant 'SS2-2' and the wild-type (WT) parent 'Sinpaldalkong2', and 75 EST clones that were highly expressed in the leaves of the SS2-2 mutant were identified. Interestingly, the expression of jasmonate (JA)-responsive genes such as vspA, vspB, and Lox2 were upregulated, whereas that of a salicylate-responsive gene PR1a was suppressed in the SS2-2 mutant. In addition, the level of JA was about two-fold higher in the leaves of the SS2-2 mutant than in those of the WT under natural growth conditions. Moreover, the JA-responsive gene expression persists in the leaves of SS2-2 mutant without rhizobia infection in the roots. Taken together, our results suggest that the nts mutation increases JA synthesis in mature leaves and consequently leads to constitutive expression of JA-responsive genes which is irrelevant to hypernodulation in the root.