• Journal of The Korean Society of Grassland and Forage Science
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• v.7 no.1
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• pp.25-30
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• 1987

There are differences in opinion as to whether nitrogen fertilizer should be used when establishing alfalfa (Medicago sativa L.). Various reports show that under a hot environment, rhizobia (Rhizobium meliloti) are not as effective in fixing atmospheric nitrogen as they are under moderate temperatures. It is also believed that the addition of nitrogen fertilizer inhibits nodulation of alfalfa seedlings. 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 grown in a hot environment. Sterile sand was used as the growing media to which a complete nutrient solution minus nitrogen was applied volumetrically to each pot daily. In addition, half of the pots received NH4-$NO_3$, at the rate of 11.2 kg per ha at seeding and at two and four weeks after planting giving a total nitrogen application rate of 33.6 kg per ha during the seven-week experimental period. 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. Plants were thinned to ten plants per pot after emergence and were grown in a controlled environment chamber with a 16-hour light period. Soil temperature at 6 cm depth ranged from 17.4^{\circ}C.$ to 31.1^{\circ}C.$ and had a daily mean of 26.5^{\circ}C.$. Plants were harvested at weekly intervals for seven weeks. Root, shoot and total length, dry weight, volume and number of nodules per plant were determined. Root, shoot, and total length were greater in seedlings grown in soil where nigrogen was applied than that grown in soil to which no nitrogen was applied. The average size of the seedlings as determined by volume and weight was more than two times greater where plants were fertilized with nitrogen. Nodule number per seedling was also greater when nitrogen was applied compared to those which received no nitrogen. The differences were greater as the plants became older. The rhizobia did not fix enough nitrogen for adequate growth of seedlings. This is probably due to high temperature growing conditions that caused the rhizobia to become relatively ineffective as compared to cooler growing conditions. Data suggests it would be desirable to apply nitrogen at seeding when alfalfa is established under hot conditions that occur in mid- or late summer.

• 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.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.313-319
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• 1995

The involvement of the cell-wall degrading enzymes in Rhizobium has long been an unsolved question about the infection process in the formation of root nodule. To assess the contribution of the cellulase to the nodulation of rhzobia, here we report the production of cellulase from R. meliloti TAL1372 which degrade carboxymethylcellulose (CMC) model substrate with CMC-plate method. We constructed a genomic library by cloning Sau3A-digested genomic DNA from R. meliloti TAL1372 into the BamHI site of the cosmid vector pLAFR3. Out of more than one thousand transductants of E. coli, one clone (pRC8-71) had CM-cellulase activity and contained pLAFR3 cosmid with 30 kb insert of R. meliloti DNA The product of CM-cellulase gene was analyzed by native PAGE. About 45 kD protein was considered to be a product of the gene. Tn5 mutagenesis reveals that the structural gene located in a ca. 3 kb KpnI fragment. The cellulase-minus mutants of R. meliloti TAL1372 were obtained by Tn5 mutagenesis of pRC8-71 and marker exchange techniques. Analyses of the nodulation ability of these Tn5 mutants showed that the CM-cellulase gene of R. meliloti TAL1372 may be involved in early nodulation development on alfalfa (Medicago satiua).

• Current Research on Agriculture and Life Sciences
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• v.12
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• pp.57-68
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• 1994

This experiment was conducted to isolate the mutants from S118 and to investigate the physiological characteristics of R. japonicum mutants. The results obtained were as follows; Based on nodulation and acetylene reduction, nodulation of rhizobia was divided into 4 groups, i.e. slow-nodulation, earlier-nodulation, infrequent-nodulation and non-nodulation. At 5% significant level, the growth of inoculated plant with SM255 was bad, but that of HP277 was good. Root-hairs curling was induced by strains S118 and HP277 on soybean, but not by strain SM255. S118 and SM255 were found to be slow-gorwers and produced alkali, whereas strain HP277 was fast-grower and produced acid in YEM broth. In litmus milk reaction, all strains indicated alkaline reaction, and serume-zone was induced weakly by HP277. All of the strains tested in this experiment utilized sucrose. HP277 and LP268 utilized xylose, whereas S118 and SM255 did not. SM255 showed bad growth in nitrogen carriers however utilization of $Ca(NO_3)_2{\cdot}4H_2O$ by HP277 was possible at 25mM and 10mM level. To compare with S118, the protein band of SM255's cell protein electrophoresis was not developed at 0.62 Rm position.

• Korean Journal of Agricultural Science
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• v.2 no.2
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• pp.309-329
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• 1975

Studies were carried out to find the effect of various soil properties on the growth, nodulation and yield using a soybean variety, 'Tongbuktae', at various soils. The results of the study are summarized as follow: 1. The formation ratios of various characters in soybeans were different before and after flowering. The root length(89.3%), stem diameter(82.1%), plant height(77.8%), number of nodes(67.4%), and number of nodes(67.9%) gave the high formation ratio before flowering. However, the formation of number of nodules(66.3%), top dry weight(74.9%) and total dry weight(71.7%)was accomplished mainly after flowering. The ratio of the formation in root dry weight was appeared to be about same in the growth period before and after flowering. 2. Nodulation ratio after flowering shelved significantly positive correlation with dry matter accumulation. 3. T/R, top dry weight/plant height and plant height/root length were significantly associated with yield. The tested soils with higher ratio mentioned above, also showed higher yield. 4. pH, oraganic matter, phosphate, potassium, total nitrogen and molybdenum in the soils were significantly correlated with nodulation and yield. Content of calcium in the soils only showed significant association with yield. 5. Soil properties influenced yield were molybdenum, calcium, organic matter, pH, number of nodules, magnesium, phosphate, total nitrogen and potassium in the order of importance. This order was varied according to the soils employed in these studies.

• Korean Journal of Soil Science and Fertilizer
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• v.26 no.3
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• pp.197-203
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• 1993

Greenhouse experiments were conducted to avaluate strain competition, nodulation, patterns of nodule occupancy and population changes of Bradyrhizobium sp. strain HCR-46 $str^{r}cep^{r}$ and CB756 $str^{r}rif^{r}$ in the rhizosphere of peanut(Arachis hypogaea L.) under different root temperatures. Inoculated with two strains using seed coating with peat slurry under different root temperatures, population of each strain in the rhizosphere increased with plant growth and multiplication rate of inoculum in the unit weight of root were showed the highest from 10 to 15days after sowing. The multiplication rate of inoculum in the rhizosphere was $28^{\circ}C$>$34^{\circ}C$>$22^{\circ}C$. The density of HCR-46 $str^{r}cep^{r}$ was more increased than that of CB756 $str^{r}rif^{r}$ under $22^{\circ}C$ and $28^{\circ}C$. While the density of two strains showed no difference under $34^{\circ}C$. Inoculated with HCR-46 $str^{r}cep^{r}$ and CB756 $str^{r}rif^{r}$, respectively at 22, 28 and $34^{\circ}C$, nodulation of each strain was dominated in its inoculation portion. Inoculated with the mixture of HCR-46 $str^{r}cep^{r}$ and CB756 $str^{r}rif^{r}$, occupancy rate of HCR-46 $str^{r}cep^{r}$ was dominated over that of CB756 $str^{r}rif^{r}$ at $22^{\circ}C$ and $28^{\circ}C$, but that was similar between them at $34^{\circ}C$. Dry mass, nodulation, nitrogen content per plant and nitrogenase activity showed higher at $28^{\circ}C$ than at $32^{\circ}C$ and $22^{\circ}C$, while those were higher in HCR-46 $str^{r}cep^{r}$ and mixing HCR-46 $str^{r}cep^{r}$ with CB756 $str^{r}rif^{r}$ than in CB756 $str^{r}rif^{r}$.

• Journal of The Korean Society of Grassland and Forage Science
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• v.14 no.3
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• pp.177-185
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• 1994

Slow seedling growth rate and nodulation failure of white clover (Trifolium repens L.) has been limited its good establishment to pastures. The experiment was done to determine the effect of removal of cotyledon and unifoliolate on the shoot, root growth, and nodule formation of 4 white clover cultivars for 8 weeks after the treatment. Four white clover cv. Regal (large leaf), Louisiana S-I (medium-large leaf), Grasslands Huia (mediumsmall leaf), and Aberystwyth S184 (small leaf), were grown in IOcm plastic pot containing 2:l:l soi1:sand:peat moss mixture until grown to cotyledon or unifoliolate stage and then removed one (Cl) or two cotyledons (C2) at cotyledon stage, and unifoliolate only (U), unifoliolate and one cotyledon (UCl) or unifoliolate and two cotyledons (UC2) at the unifoliolate stage, and the plants were sampled at 4, 6 and 8 weeks after the treatments. The intact plants had greater shoot and root dry weights, and no. of nodules than removal-treated ones. Removal treatments at cotyledon stage, the dry weight and no. of nodules more decreased in C1 and C2 than that of unifoliolate stage. While the severer cotyledon removal, the more reduction. Although the dry weights and no. of nodules steadily inclined with regrowing period, the former were higher in Regal and La. S-1 than in the others since 6 weeks after removal treatment but the latter was more in S 184 than in the others 8 weeks after removal treatment. Relationship between no. of nodules and shoot or root dry weight was analysed as linear mode while the earlier and severe removal, the steeper slope. It was concluded that severer damage of cotyledon and unifoliolate had detrimental effects on the shoot and root growth, nodule formation, and aftermath establishment of white clover.

• Molecules and Cells
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• v.27 no.2
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• pp.217-223
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• 2009

Development of symbiotic root nodules in legumes involves the induction and repression of numerous genes in conjunction with changes in the level of phytohormones. We have isolated several genes that exhibit differential expression patterns during the development of soybean nodules. One of such genes, which were repressed in mature nodules, was identified as a putative aldo/keto reductase and thus named Glycine max aldo/keto reductase 1 (GmAKR1). GmAKR1 appears to be a close relative of a yeast aldo/keto reductase YakC whose in vivo substrate has not been identified yet. The expression of GmAKR1 in soybean showed a root-specific expression pattern and inducibility by a synthetic auxin analogue 2,4-D, which appeared to be corroborated by presence of the root-specific element and the stress-response element in the promoter region. In addition, constitutive overexpression of GmAKR1 in transgenic soybean hairy roots inhibited nodule development, which suggests that it plays a negative role in the regulation of nodule development. One of the Arabidopsis orthologues of GmAKR1 is the ARF-GAP domain 2 protein, which is a potential negative regulator of vesicle trafficking; therefore GmAKR1 may have a similar function in the roots and nodules of legume plants.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.4
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• pp.400-407
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• 1989

This experiment was carried out to study the effects of nitrogen concentration of cultural solution, Rhizobium inoculation, and planting density on the growth and yield of soybean cultivars, Hwanggeumkong, Jangbaegkong, Paldalkong, Clark, and non-nodulation isoline of Clark. Rhizobium inoculation increased the stem length, particularly in Hwanggeumkong, Jangbaegkong, and decreased it significantly in non-nodulation Clark. Stem length was increased by the increase in nitrogen fertilization by the 195ppm level and decreased by the increase in plant population density. Rhizobium inoculation also increased the shoot dry weight, but significantly decreased it in non-nodulation Clark. As nitrogen concentration in the cultural solution increased the shoot dry weight decreased in Jangbaegkong and paldalkong. However, the shoot dry weight was decreased by the increase in plant population density. Rhizobium inoculation and the increase in nitrogen concentration of cultural solution increased the ratio of shoot dry weight to root weight. The Rhizobium inoculation and the increase in nitrogen concentration of cultural solution increased the grain yield per pot in Hwanggeumkong and paldolkong, While non-nodulating Clark showed significant decrease in grain yield. Grain yield per pot was also increased by the increase of plant population density. Grain yield was significantly correlated with shoot dry weight, nodule number, and nitrogen content of the soybean plants. The correlation between nitrogen contents of the soybean plants and stem length, shoot dry weight, and nodulation was significant. The allantoin-N content in stem was also significantly correlated with nodulation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.25 no.2
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• pp.38-48
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• 1980

To evaluate Rhizobium meliloti BALSAC, a strain selected from Canada as an acid tolerant one, and ordinary lucerne inoculant in acid condition, lucerne (Medicago sativa L. cv. Wairau) was inoculated and/or pelleted in the laboratory, and grown for two months in an acid soil (Lismore silt loam, pH 5.4) with three levels of lime in the, glasshouse. The results of controlled (noninoculated), nitrogen fertilized, laboratorial and commercial inoculated seeds were compared to give the following conclusions: 1. There was no significant difference in the top and root dry matter yields between two Rhizobium strains. However, Balsac inoculant showed higher single nodule dry matter weight and relatively higher number of larger nodules than the ordinary inoculant. 2. Lime application increased dry matter yields of plants and nodules, and the number of nodules per pot and the increase of nodules on the lateral roots in both inoculants. Lime application also caused an evenly distribution of nodules on the root by showing an increase of nodules mainly on the lateral roots. 3. Fertilizer nitrogen without inoculant slightly increased the nodulation percentage, the nodule dry matter weight per nodule and the relative proportion of larger nodules. 4. Commercially inoculated and pelleted seed showed less consistent results. 5. Relatively larger variations in measuring nodule characteristics was discussed and concluded that extreme cares should be given to reduce the variation.