• Title/Summary/Keyword: gnotobiotic

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Diversity of Root-Associated Paenibacillus spp. in Winter Crops from the Southern Part of Korea

  • CHEONG HOON;PARK SOO-YOUNG;RYU CHOONG-MIN;KIM JIHYUN F.;PARK SEUNG-HWAN;PARK CHANG SEUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1286-1298
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    • 2005
  • The genus Paenibacillus is a new group of bacilli separated from the genus Bacillus, and most of species have been isolated from soil. In the present study, we collected 450 spore-forming bacilli from the roots of winter crops, such as barley, wheat, onion, green onion, and Chinese cabbage, which were cultivated in the southern part of Korea. Among these 450 isolates, 104 Paenibacillus-like isolates were selected, based on their colony shape, odor, color, and endospore morphology, and 41 isolates were then finally identified as Paenibacillus spp. by 16S rDNA sequencing. Among the 41 Paenibacillus isolates, 23 were classified as P. polymyxa, a type species of the genus Paenibacillus, based on comparison of the 16S rDNA sequences with those of 32 type strains of the genus Paenibacillus from the GenBank database. Thirty-five isolates among the 41 Paenibacillus isolates exhibited antagonistic activity towards plant fungal and bacterial pathogens, whereas 24 isolates had a significant growth-enhancing effect on cucumber seedlings, when applied to the seeds. An assessment of the root-colonization capacity under gnotobiotic conditions revealed that all 41 isolates were able to colonize cucumber roots without any significant difference. Twenty-one of the Paenibacillus isolates were shown to contain the nifH gene, which is an indicator of $N_{2}$ fixation. However, the other 20 isolates, including the reference strain E681, did not incorporate the nifH gene. To investigate the diversity of the isolates, a BOX-PCR was performed, and the resulting electrophoresis patterns allowed the 41 Paenibacillus isolates to be divided into three groups (Groups A, B, and C). One group included Paenibacillus strains isolated mainly from barley or wheat, whereas the other two groups contained strains isolated from diverse plant samples. Accordingly, the present results showed that the Paenibacillus isolates collected from the rhizosphere of winter crops were diverse in their biological and genetic characteristics, and they are good candidates for further application studies.

Characterization of Plant Growth-Promoting Traits of Free-Living Diazotrophic Bacteria and Their Inoculation Effects on Growth and Nitrogen Uptake of Crop Plants

  • Islam, Md. Rashedu;Madhaiyan, M.;Boruah, Hari P.Deka;Yim, Woo-Jong;Lee, Gill-Seung;Saravanan, V.S.;Fu, Qingling;Hu, Hongqing;Sa, Tongmin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1213-1222
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    • 2009
  • The search for diverse plant growth-promoting (PGP) diazotrophic bacteria is gaining momentum as efforts are made to exploit them as biofertilizers for various economically important crops. In the present study, 17 diazotrophic strains belonging to eight different genera isolated from rice paddy fields were screened for multiple PGP traits and evaluated for their inoculation effects on canola and rice plants. All of the strains tested positive for 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity and production of indole 3-acetic acid (IAA) and ammonia ($NH_3$). Additionally, four of the strains were able to solubilize phosphorus (P), five tested positive for zinc (Zn) solubilization and sulfur (S) oxidation, and eight strains produced siderophores. Based on the presence of multiple PGP traits, 10 strains were selected for inoculation studies. Treatment with Herbaspirillum sp. RFNB26 resulted in maximum root length (54.3%), seedling vigor, and dry biomass in canola, whereas Paenibacillus sp. RFNB4 exhibited the lowest activity under gnotobiotic conditions. However, under pot culture conditions, Paenibacillus sp. RFNB4 significantly increased plant height and dry biomass production by 42.3% and 29.5%, respectively. Canola plants and rhizosphere soils inoculated with Bacillus sp. RFNB6 exhibited significantly higher nitrogenase activity. In greenhouse experiments, Serratia sp. RFNB18 increased rice plant height by 35.1%, Xanthomonas sp. RFNB24 enhanced biomass production by 84.6%, and rice rhizosphere soils inoculated with Herbaspirillum sp. RFNB26 exhibited the highest nitrogenase activity. Our findings indicate that most of the selected strains possess multiple PGP properties that significantly improve the growth parameters of the two plants when tested under controlled conditions.

Effect of Methylotrophic Bacteria in Seedling Development of Some Crops under Gnotobiotic Condition (Methylotrophic bacteria 접종이 작물 유묘 생장에 미치는 영향)

  • Hong, In-Soo;Kim, Jun-Seok;Lee, Min-Kyoung;Yim, Woo-Jong;Islam, Md. Rashedul;Boruah, Hari P. Deka;Chauhan, Puneet Singh;Han, Gwang-Hyun;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.42 no.4
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    • pp.317-322
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    • 2009
  • Healthy seedling generation is the major concern in overcoming adverse effects of biotic and abiotic stresses during tender stage of development in vegetables and horticultural crops. Because of this, priority is given to research leading to the generation of healthy seedlings in crops subjected to transplanting and bedding. In this study, growth pouch experiments were conducted to determine the effect of inoculation of six different strains of Methylobacterium sp. namely, M. oryzae CBMB20, M. phyllosphaerae CBMB27, M. suomiense CBMB120, and Methylobacterium strains CBMB12, CBMB15 and CBMB17 on the seedling development of the vegetable crops cabbage, Chinese cabbage and cucumber; and horticultural crops tomato and red pepper. Crops treated with the test strains generally showed higher seedling dry matter accumulation compared to the control. Significantly higher accumulation was exhibited by CBMB12, CBMB17, and CBMB20 in cabbage, as well as for CBMB27 and CBMB120 on tomato and Chinese cabbage, respectively. Furthermore, all the strains promoted root elongation in cucumber and tomato seedlings while in Chinese cabbage and red pepper, root elongation was observed with CBMB120 and CBMB12 inoculation, respectively. Large scale nursery study is needed to develop a thorough protocol for healthy seedling development with the use of these strains.

In vitro Evaluation of the Mechanism of Antagonism and Phosphate Solubilization by the Insect Gut Bacteria Pseudomonas sp. PRGB06 that Exhibits Plant Growth Promotion and Bio-Fertilizing Traits (배추좀나방 내장에서 분리한 식물생장촉진미생물 Pseudomonas sp. PRGB06의 길항기작과 인산가용화의 기내 평가)

  • Kim, Kyoung-A;Indiragandhi, P.;Anandham, R.;Palaniappan, P.;Trivedi, P.;Madhaiyan, M.;Han, Gwang-Hyun;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.1
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    • pp.18-25
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    • 2008
  • Pseudomonas sp. PRGB06, a bacterial strain isolated from diamondback moth (Plutella xylostella) gut, was examined for its plant growth promotion and biofertilizing traits. The bacteria growth was observed under various conditions of carbon sources, temperature, pH and salt concentrations. In addition, the mechanisms of antagonism and phosphate solubilization were investigated. The bacterial strain PRGB06, grew well using most of the tested carbon sources. The best growth was observed at $30^{\circ}C$ and pH 7. The inhibition of the pathogenic fungi was likely due to the volatile antifungal metabolite and ammonia gas produced by the bacteria. A significant positive relationship was found between the phosphate solubilization and acid production. When inoculated with PRGB06 in vitro and in gnotobiotic condition, red pepper and maize showed increase in root length, seedling vigor and dry bio-mass.

Isolation and Characterization of the IAA Producing Methylotrophic Bacteria from Phyllosphere of Rice Cultivars(Oryza sativa L.) (벼(Oryza sativa L.)의 잎 면으로부터의 IAA를 생성하는 Methylotrophic Bacteria의 분리 선별 및 특성 비교)

  • Lee, Kyu-Hoi;Munusamy , Madhaiyan;Kim, Chung-Woo;Lee, Hyoung-Seok;Selvaraj, Poonguzhali;Sa, TongMin
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.4
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    • pp.235-244
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    • 2004
  • In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf samples of 19 rice cultivars collected from three regions of Korea. Nineteen pink pigmented isolates showing characteristic growth on methanol were obtained. Physiological and biochemical characters of each isolate were examined according to methods described in Bergey's Manual of Systematic Bacteriology. When phylotypes were defined by performing numerical analysis of 37 characteristics, four distinct clusters were formed. The two reference strains, Methylobacterium extorquens AM1 and Methylobacterium fujisawaense KACC10744 were found to group under cluster IV and cluster III respectively. Cluster I diverged on the basis of nitrate reduction and four isolates showed tolerance upto 0.5 M NaCl concentrations. Two strains in cluster I and III were found to possess methane utilizing properties. Most of the isolates in all the four clusters utilized monosaccharides, disaccharide and polyols as carbon source. When the isolates were subjected for indole-3-acetic acid (IAA) analysis in the presence of L-tryptophan, only 8 isolates exhibited IAA production. In addition, the nitrogen source in the medium was found to influence the IAA production. Addition of $(NH_4)_2SO_4$ in the medium led to a 2 to 30 fold increase in the indole synthesis. However, $KNO_3$, $NH_4NO_3$ and $NH_4Cl$ substitution did not significantly stimulate the synthesis of IAA in the growth medium. Result of gnotobiotic root elongation assay significantly increased roots and shoots lengths, and number of lateral roots, which is mediated by IAA production in the culture medium. The rice seedlings primary roots from seeds treated with methylotrophic isolates were on average 27 to 56% longer than the roots from seeds treated with the uninoculated seeds. In addition, application of different high concentrations of authentic IAA ($400g\;mL^{-1}$) to roots of rice seedlings inhibited root growth. However, the IAA concentration from 10 to $200g\;mL^{-1}$, IAA promoted root growth of rice seedlings. These results suggest that bacterial IAA plays a major role in the development of the host plant root system.