• Journal of Plant Biotechnology
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• 제1권1호
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• pp.31-38
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• 1999

An artificial symbiosis was established between diazotropic Azomonas insignis and strawberry (Fragaria x ananassa). The partnership was created by in vitro techniques through callus induction and organogenesis. The basis of this partnerships is the bacterial dependence on the plants metabolic activity, using maltose in the medium as a carbon and energy source which can be utilized by the plant cells only. The presence of bacteria in the intercellular spaces of the callus tissues and regenerated plants was proven by microscopic techniques. Nitrogenase activity could also be detected in the plant tissues. For successful and high frequency introduction of bacteria to the plant tissues, biolistic gun method was used. On the basis of the DNA transfer method, Azotobacter vinelandii bacteria were delivered directly into strawberry tissues by the particle bombardment. This was the first use of living bacteria as microprojectils for bombardment of plant tissues. The treatment was successful, the presence of bacteria in the developing callus tissue and regenerated plants were detected by light and electron microscopy.

• Journal of Plant Biology
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• 제30권4호
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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.

• 생명과학회지
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• 제22권8호
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• pp.1009-1017
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• 2012

Rodobacter sphaeroides에서 orf282 유전자는 cbb3 terminal oxidase를 암호화하는 ccoNOQP 오페론과 혐기적 활성자인 FnrL을 암호화하는 fnrL 유전자 사이에 있으며, 아직은 기능이 잘 알려지지 않았다. orf282 유전자의 기능을 알기 위해 우리는 orf282의 일부를 삭제함으로써 유전자를 붕괴시켜 orf282-minus mutant를 제조하였다. 두개의 FnrL 결합 부위가 orf282의 upstream에 존재한다는 것이 밝혀져 있으며, orf282 유전자가 FnrL에 의해 양성적으로 조절된다는 것이 증명되었다. orf282 유전자는 B875와 B800-850 spectral complexes의 형성과 관련이 없다. orf282 mutant에서의 cbb3 oxidase 활성을 wild type와 비교해보면 orf282 유전자가 ccoNOQP 오페론의 조절과 cbb3 cytochrome c oxidase의 생합성과 무관하다는 것을 알 수 있다. orf282 mutant의 구조 유전자인 nifH와 조절유전자인 nifA의 프로모터 활성이 증가한 것은 orf282 유전자 산물이 nifH와 nifA의 발현에서 음성적 effector로 작용한다는 것을 시사한다.

• Applied Biological Chemistry
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• 제40권2호
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• pp.134-138
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• 1997

대기 $CO_2$ 상승시 인산공급이 대두식물체의 광합성 및 질소고정에 미치는 영향을 살펴보고저 Bradyrhizobium japonicum MN110을 접종한 대두 식물체를 인산결핍구(0.05 m-P) 및 대조구(1.0 mM-P)를 처리하여 정상 $CO_2(400\;{\mu}l/L)$와 상승 $CO_2(800\;{\mu}l/L)$의 조건인 phytotron에서 35일 동안 재배하였다. 인산결핍은 건물량을 정상 $CO_2$ 처리시 51%, 상승 $CO_2$ 처리시 64%를 감소시켰고 총 엽면적도 유의성있게 감소시켰으나 specific leaf weight는 증가시켰다. 광합성 속도 및 잎내부의 $CO_2$농도는 인산결집구에서 감소하였고 그 정도는 $CO_2$ 상승처리시 더욱 컸으며 질소고정능과 근류생체량도 인산결핍시 감소하였으나 잎의 질소농도는 인산대조구가 30% 감소하였다. 이러한 결과는 $CO_2$ 상승조건에서도 인산이 결핍될 경우 식물생육을 촉진시키지 못하며 인산은 정상 $CO_2$ 농도시 보다 $CO_2$ 상승조건에서 근류의 성장과 작용에 더욱 중요한 역할을 하는것으로 유추된다.

• The Korean Journal of Ecology
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• 제16권2호
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• pp.159-168
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• 1993

The seasonal changes of symbiotic nitrogen-fixation activity and environmental factors of autumn olive plant (Elaeagnus umbellata Thunb.), which is an important constituent species of temperate vegetation and a non-leguminous root nodule plant interacting with Frankia sp., were quantitatively analyzed inthe natural vegetations during hegrowing period. The acetylene redution April and showed two peaks of 133 and $145{\mu}M\;C_2H_4\;{\cdot}\;gfw^{-1}\;{\cdot}\;hr^{-1}$ in early June and mid September. The nitrogenase activity decreased to 10~30% during hot dry summer frommid June to the end of August, and disappeared during the dormant period of winter. The optimum rhizoshere. diurnal change showed the maximum activity in the mid-day and the minimum in the mid-night. The average contents of total nitrogen in each organ changed in the ranges of 42.5~40.1, 40.2~36.3, 30.3~28.6 and 18.4~16.2mgN $gdw^{-1}$ for nodule, leaf, root, and stem, respectively. The soil conditions of rhizosphere were weak acidic, ad seasonal variations of $NO_3^--N,\;NH_4^+-N,\;PO_4^{3-}-P$ and water contents were in the ranges of 48.3~79.5ppm,5.1~13.9ppm, 4.4~$9.9{\mu}M$ and 14.5~39.4%, respectively.

• 한국미생물·생명공학회지
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• 제13권1호
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• pp.79-85
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• 1985

대두 경남 장려품종인 장백, 광교, 강림 및 남천등의 근류로부터 Rhizobium japonicum을 분리하여 그 중 질소고정력이 R. japonicum 61A 76이나 3 I1110보다 강한 JB 101을 선별하였다. 선별된 JB 101을 N-methy1-N'-nitro-N-nitrosoguanidine 처리 및 UV조사에 많은 돌연변이체를 얻고 이들로 부터 경남 장려품종인 장백에 접종시근류 생성력 및 질소고정력등 JB 101에 비하여 훨씬 높을 뿐만아니라 SM 35에 비해서도 다소 높은 JB 65를 선별하여 이 균주의 근류생성력, 질소고정력, 고정된 암모니아가 질소고정에 미치는 영향, hydrogenase 활성등 공생관련 제 특성과 아울러 이 균주 단백질의 아미노산 조성, 2D-polyacrylamide gel을 이용한 전기영동등의 생화학적 특성도 아울러 조사하였다.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1119-1128
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• 2015

1-Aminocyclopropane-1-carboxylate (ACC) deaminase, which is encoded by some bacteria, can reduce the amount of ethylene, a root elongation inhibitor, and stimulate the growth of plants under various environmental stresses. The presence of ACC deaminase activity and the regulation of ACC in several rhizospheric bacteria have been reported. The nitrogen-fixing Pseudomonas stutzeri A1501 is capable of endophytic association with rice plants and promotes the growth of rice. However, the functional identification of ACC deaminase has not been performed. In this study, the proposed effect of ACC deaminase in P. stutzeri A1501 was investigated. Genome mining showed that P. stutzeri A1501 carries a single gene encoding ACC deaminase, designated acdS. The acdS mutant was devoid of ACC deaminase activity and was less resistant to NaCl and NiCl₂ compared with the wild-type. Furthermore, inactivation of acdS greatly impaired its nitrogenase activity under salt stress conditions. It was also observed that mutation of the acdS gene led to loss of the ability to promote the growth of rice under salt or heavy metal stress. Taken together, this study illustrates the essential role of ACC deaminase, not only in enhancing the salt or heavy metal tolerance of bacteria but also in improving the growth of plants, and provides a theoretical basis for studying the interaction between plant growth-promoting rhizobacteria and plants.

• 미생물학회지
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• 제24권3호
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• pp.211-220
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• 1986

Nitrogen fixation (nif) genes of Rhizobium leguminosarum were transferred to nif Klebsiella pneumoniae and E. coli by conjugation after partial heat induction of $RP_4$ :: Mu cts in Rhizobium $R^+$ transconjugant, and the hybrid plasmids in the transconjugant strains were isolated and characterized. In order to transfer the nif genes from Rhizobium, the hybrid plasmid $RP_4$ :: Mu cts was transferred by conjugation from E. coil to the symbiotic nitrogen fixer, R. leguminosarum. After stabillity test, the $RP_4$ :: Mu cts in Rhixobium $R^+$ transconjugant was subjected to partial heat induction by culturing it statically at $38^{\circ}C$ for 16 hours, and then conjugated with the nif defective mutant strains of K. pneumoniae or nif mutant strains of E. coli having whole nif gene plasmid. Recombinant strains of K. pneumoniae, which could grow in a N-free medium and exhibit the nitrogenase activity were selected. However, in the case of E. coli, they could grow well in a NA medium containing antibiotices, but hardly frow in a N-free medium. The hybrid plasmids in these transconjugal strains were isolated by gel electrophoresis and compared their molecular sizes.

• 한국작물학회지
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• 제43권1호
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• pp.23-27
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• 1998

Inhibition of nodule formation and nitrogen fixation by soil nitrogen, primarily nitrate, is well known in legume plants. The present study was undertaken to evaluate the effect of ${NO_3}^-$ on the nodulation, nitrogenase activity, and growth of supernodulating soybean mutant and its wild type. A greenhouse study was conducted to compare two of supernodulating mutants, 'SS2-2' and 'nts 382', with the normal nodulating cultivar 'Sinpaldalkong 2' when grown in a 1-l styroform cup filled with sand, and fertilized with five levels of ${NO_3}^-$ (0, 2, 4, 8, and 12 mM). During the growth period, each plant was supplied two or three times a week with 50 mL of nutrient solution. Supernodulating soybean mutants, SS2-2 and nts 382, showed more nodules and nodule mass, and greater $C_2\;H_2$ activity than the wild type, Sinpaldalkong 2, regardless of the level of exogeneous nitrogen supply. On the other hand, total dry weight of SS2-2 mutant, which was smaller than Sinpaldalkong 2, did not respond to the various ${NO_3}^-$-N levels. This suggested that supernodulating SS2-2 mutant could maintain fairly high total dry weight at the low ${NO_3}^-$-N level, even in the absence of exogeneous ${NO_3}^-$-N in the nutrient solution. From the reduced top growth and high nitrogen fixing ability of supernodulating mutants, it was surmised that supernodulating mutant could potentially protect agricultural environments from pollution through the reduction in nitrogen fertilization as well as maintain fairly high yield with increasing planting density.

• 미생물학회지
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• 제32권2호
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• pp.97-101
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• 1994

경기도, 충청남도 일원의 벼 근권에서 400~900nmol $C_2H_4/hr/vial$의 아세틸린 환원 활성을 갖는 질소고정균인 Azospirillum 15균주를 분리하였다. 이 분리균주들은 $1.0{\times}3.0{\mu}m$크기를 갖는 vibrioid 형태이었으며 액체배양시 monopolar single 편모를 가졌다. 이 분리균주들은 생리적, 형태적 특성에 따라 두 그룹으로 분류되었는데, 그룹 I 균주들은 탄소원으로 glucose를 잘 이용하였고 biotin 요구성인 반면, 그룹 II 균주들은 glucose를 전혀 이용하지 못하였고 biotin 비요구성이었다. 또한 semisolid 무질소 배지에서 48시간 배양할 때 그룹 I 균주들은 원래의 vibrioid 형태에서 좀 더 길어지면서 S자 모양으로 변하는 pleomorphic 특성을 보인 반면, 그룹 II 균주들은 원래의 모양과 운동성을 유지하였다. 이상의 결과로부터 그룹 I 균주들은 Azospirillum lipoferum으로, 그룹 II 균주들은 Azospirillum brasilense으로 동정하였다.