• Journal of Plant Biology
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• v.30 no.4
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• pp.257-266
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• 1987

Investigation on the ability of nitrogen usage by N2-fixing Nostoc muscorum and cultured tobacco cells when they were associately cultured on nitrogen-free media was carried out. Also, effect of polyamines on the associated cultured condition was carried out. In addition, morphological changes of N. muscorum and cultured tobacco cells in associate culture were observed to detect the possibility of induction of nitrogen fixing ability on cultued plant cells. The activity of nitrogenase increased markedly when N. muscorum was grown exclusively on nitrogen-free media. When N. muscorum was cultured associately with cultured tobacco cells on nitrogen-free media containing polyamines, high activity was detected in 10-4 M spermine treated group. Investigation on the change of polyamine amounts showed two times increase in spermidine and eight times increase in spermine on a associate culture. These effects of associated culture were shown through morphological change such as dense loclization of N. muscorum around the cultured tobacco cells as well as inside the cells. These results indicate the viability of N. muscorum in cultured tobacco cells and possible induction of nitrogen fixation ability by symbiosis.

• Journal of Plant Biology
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• v.30 no.3
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• pp.205-213
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• 1987

Soybean, inoculated with effective Rhizobium japonicum 110, were grown by sand culture with nutrient solution containing either of 0, 1, 3, 10 or 30mM NO3-/l, and analyzed growth characteristics, NR activity, N2-fixation activity, and changes of ureide contents during the growing period. The amount of nodule formation decreased abruptly by nitrate treatment, the maximum nodule dry weight was 1.59, 1.05, 0.78, 0.09 and 0.008 g plant-1, respectively for each treatment on the 98th day. Specfic activity of N2-fixation showed the maximum rates of 140, 101, 37, 5 and 2.2 nM dw.mg-1.hr-1, respectively for each treatment in the earlier growth period. The maximum acetylene reduction activity on the 98th day after sowing was 81.5, 35.3, 14.3, 0.1 and 0.0045 $\mu$M C2H4 plant-1.hr-1, respectively for 0, 1, 3, 10 and 30 mM of NO3- gradients. Nitrate reduction activity increased along with nitrate gradients, and decreased abruptly with age. Relative abundance of ureides in plant organs was high in reproductive growth, and showed the maximum value in fully symbiotic dependent plant. Relative abundance of ureides in stem is a useful indication for the evaluation of nitrogen fixation in nodules of symbiotic plant.

• Journal of Ecology and Environment
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• v.34 no.3
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• pp.279-286
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• 2011

The effect of allelochemicals on growth, root nodule nitrogen fixation activity, and ion patterns of soybeans were investigated. We prepared 50 g/L (T50), 100 g/L (T100), and 200 g/L (T200) extract concentrations by soaking fresh red pine needles in a nutrient solution. Adding needles to the nutrient solution increased the content of total phenolic acids, osmolality, and total ions. The total phenolic content in the T50, T100, and T200 extracts were $206{\pm}12.61$, $335{\pm}24.16$, and $603{\pm}12.30$ mg gallic acid equivalents, respectively. The $K^+$, $Mg^{2+}$, $Ca^{2+}$, and $PO_4^{3-}$ content increased by adding needles to the nutrient solutions, whereas $SO_4^{2-}$ content decreased. The growth inhibition of soybeans was proportional to the needle extract concentrations, and the T100 and T200 concentrations resulted in remarkable growth inhibition. On day 20 after treatment, dry weight and nitrogen fixation activity of the root nodules were reduced by the T100 and T200 treatments, whereas the T50 treatment was not difference from the control. After day 10, total ion content in all treatment groups was not different in comparison with the control. However, total ionic content in all treatment groups decreased significantly compared with that in the control after day 20. The lowest total ion value was found for the T200 concentration. The T200 treatment also resulted in significantly reduced $SO_4^{2-}$ content. The amounts of $Mg^{2+}$, $Ca^{2+}$, and $Mn^{2+}$ were higher than those of the control for the T50 treatment on day 10 and for T100 on day 20 after treatment. A significant increase in osmolality was observed in the T200 treatment on day 10 and in the T100 treatment on day 20. These results suggest that under severe allelochemical stress conditions, a remarkable reduction in nodule formation, nitrogen fixation activity, and ion uptake eventually resulted in a decrease in leaf production. Furthermore, increased $K^+$, $Mg^{2+}$, $Ca^{2+}$, $Mn^{2+}$, and osmolality in soybeans exposed to lower concentrations of allelochemicals than the critical stress level helped overcome the stress.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1169-1179
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• 2020

In this study, two soybean genotypes, i.e., aluminum-tolerant Baxi 10 (BX10) and aluminumsensitive Bendi 2 (BD2), were used as plant materials and acidic red soil was used as growth medium. The soil layers from the inside to the outside of the root are: rhizospheric soil after washing (WRH), rhizospheric soil after brushing (BRH) and rhizospheric soil at two sides (SRH), respectively. The rhizosphere bacterial communities were analyzed by high-throughput sequencing of V4 hypervariable regions of 16S rRNA gene amplicons via Illumina MiSeq. The results of alpha diversity analysis showed that the BRH and SRH of BX10 were significantly lower in community richness than that of BD2, while the WRH exhibited no significant difference between BX10 and BD2. Among the three sampling compartments of the same soybean genotype, WRH had the lowest community richness and diversity while showing the highest coverage. Beta diversity analysis results displayed no significant difference for any compartment between the two genotypes, or among the three different sampling compartments for any same soybean genotype. However, the relative abundance of major bacterial taxa, specifically nitrogen-fixing and/or aluminum-tolerant bacteria, was significantly different in the compartments of the BRH and/or SRH at phylum and genus levels, indicating genotype-dependent variations in rhizosphere bacterial communities. Strikingly, as compared with BRH and SRH, the WRH within the same genotype (BX10 or BD2) always had an enrichment effect on rhizosphere bacteria associated with nitrogen fixation.

• Journal of Plant Biology
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• v.29 no.1
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• pp.67-75
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• 1986

An experimental system for the possibility of association between cultured plant cells and a cyanobacterium (N. muscorum) was investigated. The association was established in nitrogen-free medium, and nitrogen source was provided by N. muscorum through its own nitrogen fixation. N. muscorum was distributed on the outer part and intercellular spaces of the cultured plant cells. Polyamines were treated to the nitrogen-free medium to improve the association. Polyamines increased nitrogenase activity and protein amount in the association between N. muscorum and the cultured plant cells, as compared to the association without polyamines. Especially, spermine showed an increasing effect on the nitrogenase activity and the protein content in the association of tobacco and N. muscorum, while spermidine showed similar effect in case of soybean and N. muscorum.

• Journal of Microbiology and Biotechnology
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• v.21 no.2
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• pp.136-146
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• 2011

This study provides first-hand proteomic data on the survival strategy of Anabaena sp. PCC 7120 when subjected to long-term iron-starvation conditions. 2D-gel electrophoresis followed by MALDI-TOF/MS analysis of iron-deficient Anabaena revealed significant and reproducible alterations in ten proteins, of which six are associated with photosynthesis and respiration, three with the antioxidative defense system, and the last, hypothetical protein all1861, conceivably connected with iron homeostasis. Iron-starved Anabaena registered a reduction in growth, photosynthetic pigments, PSI, PSII, whole-chain electron transport, carbon and nitrogen fixation, and ATP and NADPH content. The kinetics of hypothetical protein all1861 expression, with no change in expression until day 3, maximum expression on the $7^{th}$ day, and a decline in expression from the $15^{th}$ day onward, coupled with in silico analysis, suggested its role in iron sequestration and homeostasis. Interestingly, the up-regulated FBP-aldolase, Mn/Fe-SOD, and all1861 all appear to assist the survival of Anabeana subjected to iron-starvation conditions. Furthermore, the $N_2$-fixation capabilities of the iron-starved Anabaena encourage us to recommend its application as a biofertilizer, particularly in iron-limited paddy soils.

• Korean Journal of Ecology and Environment
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• v.38 no.4 s.114
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• pp.510-517
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• 2005

The filamentous cyanobacterium Anabaena is capable of both photosynthesis and nitrogen fixation which probably facilitated its incredible adaptation and proliferation in freshwater environments. A small gene, patS, was found to block nitrogen fixing cells from developing which resulted in death of Anabaena in the absence of combined nitrogen sources. We analyzed the DNA sequences in the vicinity of the patS gene by using a codon usage program and detected no codon bias other than the patS open reading frame. Three overlapping cosmids that contain the patS gene were identified, and the presence of other known heterocyst-controlling genes was examined. The patS expression in response to nitrogen starvation was analyzed at the level of transcription and translation by using Northern blot analyses and lacZ-reporter-gene fusion experiments, respectively. The patS expression increased rapidly (within 12 hours) upon the removal of combined nitrogen from the media.

• Journal of Plant Biology
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• v.35 no.4
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• pp.317-322
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• 1992

The nitrogen fixation activity and environmental factors of Lespedeza bicli!oy Turcz, forming annual root nodules by symbiotic Rhizobium sp. were analyzed in the field conditions during the growing period. Seasunal changes of $N_2-fixation$ activity showed the maximum value of $120\;\mu\textrm{M}\;C_2_H4{\cdot}noduie\;g\;fw^{-1}{\cdot}hr^{-1}$ during the active growing period (June) and varied significantly depending on the growth phase and environmental factors. The maximum activities were attained at the conditions of pH 7, $30^{\circ}C$ of temperature, 18 Kpa of oxygen partial pressure and inhibited by water stress and nitrogen sources. The habitat soil was weak acidic and poor in nitrogen, phosphorus and organic matter contents. The leaf area ratios and chlorophyll contents were ranged from 442 to $48;\textrm{cm}^2{\cdot}g\;dw^{-1}$ and from 33 to $38\;\mu\textrm{g}\;chI{\cdot}\textrm{cm}^2$. Nitrogen contents in each organ showed the maximum of 46, 19 and $11\;mg{\cdot}g\;dw^{-1}, respectively for leaf, rool and stem in the early period. The highest phosphorus contents were 4.2, 1.2 and $0.6\;mg{\cdot}g\;dw^{-1}$, respectively for leaf. root and stem in early growing period. The allocation ratios of nitrogen quantity showed 60% for leaves and 73% for roots during the active and late growth period, and 22% [or stems in average. The allocalion ralios o[ phosphorus quantity showed 58% for leaves during the most productive period, 70% for roots in the pre-growth stage and 26% for stems in average.verage.

• Journal of Plant Biology
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• v.25 no.4
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• pp.153-160
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• 1982

The kinetics of $^{14}C$ fixation have been investigated in Anabaena flos-aquae growing on ${NH}_4+$,$NO_3-$ and $NO_2-N$ in batch cultures. Growth rate was highest with ${NH}_4+$, followed by $NO_2-$ and finally $NO_2$. The compensation intensity($I_0$) and the half-saturation irradiance($K_1$) with $K_1$ were higher than with other N sources, but the maximum C fixation rate($P_{max}$) was lower. The ($P_{max}$)/$K_1$ ratio, which is analogous to quantum efficiency at low irradiance ranges, was also lower with $N_2$. All these parameters except $K_1$ decrease with culture age, or decreasing growth rate. Since $^{14}C$ uptake measures net photosynthesis, the higher values of $I_0$ and $K_1$, and the low values of $P_{max}$/$K_1$ ratio with $N_2$ appear to be related to the high energy demand of $N_2$fixation. They may also be related to the lox maximum growth rate with $NO_2-N$.

• Journal of Plant Biology
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• v.34 no.1
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• pp.31-35
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• 1991

Induction of symbiosis between N. muscorum and cultured tobacco cells associately cultured on nitrogen-free medium and effects of polyamines on culture condition were carried out. Analysis of amino acid composition in associate cultures showed increase in total amino acid amounts than single culture of cultured tobacco cells and methionine was markly increased in associative culture on N-free medium treated with 1 mM spermine. These results indicated that the composition of amino acids increased effectively in associative cultures by nitrogen fixation of N. muscorum.scorum.