• Journal of Microbiology
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• v.40 no.3
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• pp.205-210
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• 2002

To study interactions between $C_{4}$-dicarboxylic acid transport protein D and E$\sigma$$^{54}$ in the dctA promoter regulatory region, we used the challenge phage system. An ant＇-｀lac fusion was recombined onto the challenge phage, and this ant＇-｀lac fusion along with Pant and the R. meliloti dctA promoter regulatory region were cloned onto a plasmid. The plasmid bearing the ant＇-｀lac fusion was used as a reporter plasmid in a coupled transcription-translation system. Addition of purified $\sigma$$^{54}$ to the coupled system specifically repressed transcription of the plasmid-borne ant＇-｀lac fusion. When DCTD was added along with $\sigma$$^{54}$ to the coupled system, transcription of the ant＇-｀lac fusion was even further repressed, suggesting that DCTD may stabilize closed complexes between E$\sigma$$^{54}$ and the dctA promoter.

• 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.

• 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.

• Biomedical Science Letters
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• v.6 no.1
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• pp.1-9
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• 2000

To identity the genes responsible for the biosynthesis of exopolysaccharide, recombinant plasmids pUEX10 and pLEX10 were constructed from plasmid pLEX3 which was isolated from the recombinant cosmid library of Zoogloea ramigera 115. The complete nucleotide sequence of the 1.7 kb genomic DNA insert in plasmid pUEX10 was determined. Its analysis identified two open reading frames (ORF3 & ORF4) which could encode two proteins. The amino acid sequence derived from ORF3 showed the homology with gumC protein in Xanthomonas campestris as well as exoP protein in Rhizobium melizoti. The partial amino acid sequence of ORF4 showed the homology with polysaccharide export protein in Thermotoga maritima. Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 showed the similar pattern for EPS production. Yield of exopolysaccharides produced by Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 was 0.26% (w/v) and 0.16% (w/v), respectively.

• 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.