• Title/Summary/Keyword: nitrogen-fixing bacteria

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Genetic and Phenotypic Diversity of Plant Growth Promoting Rhizobacteria Isolated from Sugarcane Plants Growing in Pakistan

  • Mehnaz, Samina;Baig, Deeba N.;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1614-1623
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    • 2010
  • Bacteria were isolated from roots of sugarcane varieties grown in the fields of Punjab. They were identified by using API20E/NE bacterial identification kits and from sequences of 16S rRNA and amplicons of the cpn60 gene. The majority of bacteria were found to belong to the genera of Enterobacter, Pseudomonas, and Klebsiella, but members of genera Azospirillum, Rhizobium, Rahnella, Delftia, Caulobacter, Pannonibacter, Xanthomonas, and Stenotrophomonas were also found. The community, however, was dominated by members of the Pseudomonadaceae and Enterobacteriaceae, as representatives of these genera were found in samples from every variety and location examined. All isolates were tested for the presence of five enzymes and seven factors known to be associated with plant growth promotion. Ten isolates showed lipase activity and eight were positive for protease activity. Cellulase, chitinase, and pectinase were not detected in any strain. Nine strains showed nitrogen fixing ability (acetylene reduction assay) and 26 were capable of solubilizing phosphate. In the presence of 100 mg/l tryptophan, all strains except one produced indole acetic acid in the growth medium. All isolates were positive for ACC deaminase activity. Six strains produced homoserine lactones and three produced HCN and hexamate type siderophores. One isolate was capable of inhibiting the growth of 24 pathogenic fungal strains of Colletotrichum, Fusarium, Pythium, and Rhizoctonia spp. In tests of their abilities to grow under a range of temperature, pH, and NaCl concentrations, all isolates grew well on plates with 3% NaCl and most of them grew well at 4 to $41^{\circ}C$ and at pH 11.

Associated Nitrogen Fixation in the Rhizosphere of Rice in Saline and Reclaimed Saline Paddy Soil -IV. Chemotaxis of Associative N2-fixing Bacteria to Single or Combined Sugars and Root Exudates (간척지토양(干拓地土壤)의 수도근권(水稻根圈)에서 협생질소고정(協生窒素固定)에 관(關)한 연구(硏究) -제사보(第四報). 협생질소고정균(協生窒素固定菌)의 단당(單糖) 및 복합당류(複合糖類)와 근분비물질(根分泌物質)에 대(對)한 화학주화성(化學走化性))

  • Lee, Sang-Kyu;Suh, Jang-Sun;Ko, Jae-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.4
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    • pp.359-367
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    • 1987
  • A method for evaluating bacterial chemotactic responses toward several single of combined sugars and sterile mucilage from the different rice cultivars had been tested. Bacterial genus of Azospirillum, Pseudomonas and Agrobacterium were specially identified from the histosphere of different rice cultivars and graminea grasses in saline and reclaimed saline paddy soil. To evaluate chemotaxis of these strains a modification of Fendrik channel method was used. Under this condition Azospirillum lipoferum Ecc 3-1 reacted stereoisomerically fomulating the single migration ring while Agrobacterium radiobacter Ecc 1-1 and Pseudomonas sp Ecc 4-1 did not. Strains specificities of chemotaxis to the single sugar such as D(+)-glucose and D(+)-fructose were less prominent than malic and citric acid. Chemotactic responses to the combined sugar such as D-galacturonic acid and the L-aspartate were found high attracting reaction than other combined sugars. Chemotaxis of associative $N_2$-fixing bacteria to the root exudates of different rice cultivars were differed among bacterial strains and rice cultivars.

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The Hypernodulating nts Mutation Induces Jasmonate Synthetic Pathway in Soybean Leaves

  • Seo, Hak Soo;Li, Jinjie;Lee, Sun-Young;Yu, Jae-Woong;Kim, Kil-Hyun;Lee, Suk-Ha;Lee, In-Jung;Paek, Nam-Chon
    • 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.

Polyphasic Analysis of the Bacterial Community in the Rhizosphere and Roots of Cyperus rotundus L. Grown in a Petroleum-Contaminated Soil

  • Jurelevicius, Diogo;Korenblum, Elisa;Casella, Renata;Vital, Ronalt Leite;Seldin, Lucy
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.862-870
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    • 2010
  • Cyperus rotundus L. is a perennial herb that was found to be dominating an area in northeast Brazil previously contaminated with petroleum. In order to increase our knowledge of microorganism-plant interactions in phytoremediation, the bacterial community present in the rhizosphere and roots of C. rotundus was evaluated by culture-dependent and molecular approaches. PCR-DGGE analysis based on the 16S rRNA gene showed that the bacterial community in bulk soil, rhizosphere, and root samples had a high degree of similarity. A complex population of alkane-utilizing bacteria and a variable nitrogen-fixing population were observed via PCR-DGGE analysis of alkB and nifH genes, respectively. In addition, two clone libraries were generated from alkB fragments obtained by PCR of bulk and rhizosphere soil DNA samples. Statistical analyses of these libraries showed that the compositions of their respective populations were different in terms of alkB gene sequences. Using culturedependent techniques, 209 bacterial strains were isolated from the rhizosphere and rhizoplane/roots of C. rotundus. Dot-blot analysis showed that 17 strains contained both alkB and nifH gene sequences. Partial 16S rRNA gene sequencing revealed that these strains are affiliated with the genera Bosea, Cupriavidus, Enterobacter, Gordonia, Mycoplana, Pandoraea, Pseudomonas, Rhizobium, and Rhodococcus. These isolates can be considered to have great potential for the phytoremediation of soil with C. rotundus in this tropical soil area.

Impact of a Glyphosate-Tolerant Soybean Line on the Rhizobacteria, Revealed by Illumina MiSeq

  • Lu, Gui-Hua;Zhu, Yin-Ling;Kong, Ling-Ru;Cheng, Jing;Tang, Cheng-Yi;Hua, Xiao-Mei;Meng, Fan-Fan;Pang, Yan-Jun;Yang, Rong-Wu;Qi, Jin-Liang;Yang, Yong-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.561-572
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    • 2017
  • The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.

Characterization of Rhizobacteria Isolated from Family Solanaceae Plants in Dokdo Island (독도에 서식하는 가지과식물로부터 분리된 근권세균의 특성)

  • Ham, Mi-Seon;Park, Yu-Mi;Sung, Hye-Ri;Sumayo, Marilyn;Ryu, Choong-Min;Park, Seung-Hwan;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.110-117
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    • 2009
  • To characterize plant root-associated bacteria in wild plant family Solanaceae, Solanum nigrum L. plants were collected in Dokdo island. Forty four strains of nitrogen-fixing or spore-forming bacteria were isolated from rhizosphere of Solanum nigrum L. plants. Among these, 19 strains were able to produce auxin. Thirteen strains of these produced siderophore as determined by color reaction on CAS-blue plate, 8 strains were able to solubilize phosphate. The 16S rDNA genes of the isolated bacteria were amplified and sequenced. Model plants, pepper and tobacco, were established in order to evaluate the bacterial capacities eliciting growth promotion and induced systemic resistance. The plants treated with strain KUDC1009 were more resistant and capable of growth-promotion than control plants when challenged by either Xanthomonas axonopodis pv. vesicatoria or Erwinia carotovora sub. carotovora strain SCC1. Rhizobacteria isolated from Dokdo island can promote growth of wild type Solanum nigrum L. under much environmental stresses.

The Effect of Cold-adaptation on Stress Responses and Identification of a Cold Shock Gene, capA in Bradyrhizobium japonicum (Bradyrhizobium japonicum의 저온 전처리에 의한 환경스트레스 내성 증진에 대한 연구)

  • 유지철;노재상;오은택;소재성
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.45-49
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    • 2002
  • Bradyrhizobium japonicum is a soil bacterium with a unique ability to infect the roots of leguminous plants and establish a nitrogen-fixing symbiosis, which has been used as a microbial manure. In this study, we examined the stress response after pretreatment of cells with cold temperature. When pre-treated with cold temperature ($4^{\circ}C$) for 16 hr, B. japonicum increased the viability in subsequent stress-conditions such as alcohol, $H_2O_2$, heat, and dehydration. For cold adpatation, cultured B. japonicum was exposed to $4^{\circ}C$. Upon subsequent exposure to various conditions, the number of adapted cells pretreated by cold adaptation was 10-1000 fold higher than that of non-adaptated ones. It appeared de novo protein synthesis occurred during adaptation, because a protein synthesis inhibitor, chloramphenicol abolished the increased stress tolerance. By using a degenerate PCR primer set, a csp homolog was amplified from B. japonicum genome and sequenced. The deduced partial amino acid sequence of the putative Csp (Cold shock protein) shares a significant similarity with known Csp proteins of other bacteria.

Effects of Mixed Planting of Green Manure Crops Supplemented with Humic Substance on the Biological Soil Health Indicators of Reclaimed Soils (녹비식물 혼합재배에 휴믹물질 투입이 정화처리토양의 생물학적 토양 건강성 지표에 미치는 영향)

  • Bae, Bumhan;Park, Hyesun;Kang, Sua
    • Journal of Soil and Groundwater Environment
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    • v.26 no.5
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    • pp.49-59
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    • 2021
  • The effects of green manure crops, hairy vetch and sesban, supplemented with HS (humic substance) on biological soil health indicators was studied in a pot containing two kinds of reclaimed soil previously contaminated with petroleum hydrocarbons; a soil remediated by land-farming (DDC) and another soil by low-temperature thermal desorption (YJ). Treatments include no plant (C), plants only (H), and plants+2% HS (PH), which were evaluated in a pot containing respective soil. Biological indicators include microbial community analysis as well as soil enzyme activities of dehydrogenase, 𝛽-glucosidase, N-acetyl-𝛽-D-glucosaminidase (NAG), acid/alkaline phosphatase, arylsulfatase, and urease. Results showed an increase of enzyme activities in pot soils with plants and even greater in soils with plants+HS. The enzyme activities of DDC soil with plants (DDC_P) and with plants+HS (DDC_PH) increased 1.6 and 3.9 times on average, respectively than those in the control. The enzyme activities YJ soil with plants (YJ_P) and with plant+HS (YJ_PH) increased 1.8 and 3.8 times on average, respectively than those in the control. According to microbial community analysis, the relative abundance of nitrogen-fixing bacteria in DDC and YJ soil was increased from 1.5% to 7% and from 0 to 5%, respectively, after planting hairy vetch and sesban. This study showed that mixed planting of green manure crops with a supplement of humic substance is highly effective for the restoration of biological health indicators of reclaimed soils.

Effects of Charcoal Powder on the Growth and Development of Red Pepper and the Changes of Soil Microflora (목탄분말 시용이 고추의 생육 및 토양미생물상 변화에 미치는 영향)

  • 김승환;이상민;이윤정;김한명;송석용;송범헌
    • Korean Journal of Organic Agriculture
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    • v.11 no.3
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    • pp.55-64
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    • 2003
  • It was investigated the possibility to use charcoal powder as beneficial soil conditioner, which used frequently in environmentally friendly agricultural farming system. For this purpose, the effects of charcoal powder on the growth of red pepper and chemical and microbiological properties of soil were also determined. The application of charcoal powder resulted in no significant differences of pH and EC in the soil compared to those of control. However, small particle size of charcoal powder increased yield of red pepper while large charcoal powder resulted in decrease of root growth of red pepper. Furthermore, the application of charcoal powder resulted in changes of soil microflora relating to plant growth stage. The number of the nitrogen fixing bacteria and fungi increased at the early growth stage, while phosphate releasing fungi in the soil increased at the late stage of growth by charcoal powder application. These beneficial effect of charcoal powder on the soil microbial properties was larger by the use of smaller particle size of charcoal powder. Therefore, it indicated that the small size of charcoal powder might be more influential on the red pepper yield and soil microbial properties may be due to large capacity of nutrients uptake for the plant and microorganisms. Additionally the optimal application amount of charcoal powder for the red pepper could be suggested as much as 300kg 10a$^{-1}$ for the both purposes of improvement of crop yield and retardation of the nutrients accumulation by excess charcoal application.

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Isolation of Pea and Soybean Nodule Bacteria and Assessment of their Nitrogen-fixing Capacities (완두 및 대두근류균(根瘤菌)의 분리(分離) 및 질소고정능력(窒素固定能力)의 비교(比較))

  • Kim, Seung-Yeol;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.5 no.2
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    • pp.136-144
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    • 1978
  • A series of experiments over three years was planned for practical application of rhizobia in farms and grass lands in Korea. This is the report for the studies of the first year mainly on the isolation and characterization of rhizobial strains, and on the assessment of their nodulation abilities and nitrogen fixation capacities. 1. Total number of 88 strains for soybean group and 22 st ra ins for pea and vetch group was isolated from nodules which were taken from legumes grown in Daekwanyong, Cheju and various places in Korea. 2. Morphological and cultural characteristics of the strains were studied, and attempts were also made to investigate their antigenic properties and to demonstrate lysogenic strains in these groups. The results were : i) the isolates varied in cultural characteristics on yeast mannitol broth and agar, and in degree of congo reel absorption ; ii) similarities in their antigenic properties were found between/among the strains: G-3/G-9/D216, G-20/G-52 in soybean group; iii) no lysogeny was found in the strains of these groups. 3. Plant infection tests by test tube and bottle method in light room were carried out to elucidate the ability of the strains to nodulate specific legumes and of the capacity of such nodules to fix atmospheric nitrogen. The isolates were grouped into non- invasive, ineffective, or effective to the legumes. Those strains which produced effective nodules, supporting similar level of growth as nitrate control, were: P-3, 4 and 8 in pea and vetch group; G-23, 27, and, D-216 in soybean group.

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