• Title/Summary/Keyword: endophytic bacteria

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Isolation of Nitrogen-Fixing Bacteria from Gramineous Crops and Measurement of Nitrogenase Activity (벼과식물로부터 질소고정균의 분리와 Nitrogenase 활성 측정)

  • 최은화;이상은;윤기순;권덕기;손재근;박승환;한명숙;김사열
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.18-24
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    • 2003
  • For researching nitrogen-fixing bacteria associated with gramineous crops, we collected growing roots of rices, wheats, oats, barleys, ryes, and maizes at 19 sites of southern Korean peninsula. Endophytes and free living bacteria were isolated from those crop roots. Sixty-three isolates were classified on the basis of different morphology, size, color, host of colony, and the 16S rDNAs sequence. The analyses of PCR amplification for nifH gene and nitrogenase activity assay, revealed that all isolates contained nitrogen-fixing abilities. In addition, most of them have cellulase activity which is one of the common features of endophytic bacteria from plant.

Biocontrol Efficacy of Endophytic Bacteria Flavobacterium hercynim EPB-C313 for Control of Chinese Cabbage Clubroot (Flavobacterium hercynium EPB-C313 균주를 이용한 배추 뿌리혹병 생물적 방제)

  • Hahm, Soo-Sang;Kim, Jong-Tae;Han, Kwang-Seop;Kim, Byung-Ryun;Kim, Hong-Kyu;Nam, Yun-Kyu;Yu, Seung-Hun
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.210-216
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    • 2012
  • Clubroot of Chinese cabbage by Plasmodiophora brassicae, was found to be high virulent to the Chinese cabbage, turnips and cabbage. It this study, the endophytic bacteria Flavobacterium hercynium EPB-C313, which was isolated from tissues of Chinese cabbage, was investigated the antimicrobial activity on the inactivation of resting spores and its control effect on clubroot disease by bioassay. The bacterial cells, culture solutions, and culture filtrates of F. hercynium EPB-C313 inactivate the resting spores of P. brassicae with 90.4, 36.8, and 26.0%, respectively. The clubroot was inhibited with 100% by dipping the seedlings of Chinese cabbage in culture solutions of F. hercynium EPB-C313 before planting, however the chemmical 'fluazinam' was 91.7% in pot tests. Complex treatment were highly enhanced control efficacy with 63.7% at field of 50% diseased plants by soil incorporation with the pellet contains organic matter and F. hercynium EPB-C313, seedling drench of culture solution of F. hercynium EPB-C313 and soil drench with 10 days after planting. These results imply that the F. hercynium EPB-C313 is a very useful biological control agent of clubroot disease of Chinese cabbage.

Isolation and Characterization of High Viscosity Polysaccharide Producing Endophytic Bacteria from Pueraria Root (고점도 다당류를 생산하는 갈근 내생균의 분리 및 특성)

  • Whang, Kyung-Sook;Choi, Seung-Hyun;Han, Song-Ih
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.341-345
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    • 2007
  • Fifty endophytic bacteria, which produced slime around the colonies, were isolated from Pueraria roots. In particular, HDN-14, TDG-3, and TNB-3 strains, which appeared to be high viscosity producers, were selected. These strains produced high levels of polysaccharides in Puerara root medium extract. The purified polysaccharide was digested with 1N HCI and analyzed by HPLC, with glucose ($45.6{\sim}63.1%$), maltose ($14.6{\sim}23.7%$), and fructose ($17.4{\sim}23.7%$) detected as constitutive sugars. When determined by the homology relationship of the 16S rDNA sequence with the relative taxa, the HDN-14 and TNB-3 strains were closely ($99.06{\sim}99.32%$) related to the Pseudomonas $koreensis^T$ and Pseudomonas $jessenii^T$, while TDG-3 were closely ($99.48{\sim}99.74%$) related to Pseudomonas $plecoglossicida^T$, Pseudomonas $mosselii^T$, and Pseudomonas $monteilii^T$. The major cellular Pseudomonas acids are $3OH-C_{10:0}$, $2OH-C_{12:0}$, $3OH-C_{12:0}$, and $3OH-C_{12:1}$, with these strains being further differentiated in species belonging to the genus Pseudomonas.

Crossbreeding and parental lineage influences the diversity and community structure of rice seed endophytes

  • Walitang, Denver I.;Halim, MD Abdul;Kang, Yeongyeong;Kim, Yongheon;Sa, Tongmin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.161-161
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    • 2017
  • Seed endophytes are very remarkable groups of bacteria for their unique abilities of being vertically transmitted and conserved. As plants attain hybrid vigor and heterosis in the process of crossbreeding, this might also lead to the changes in the community structure and diversity of plant endophytes in the hybrid plants ultimately affecting the endophytes of the seeds. It would be interesting to characterize how seed endophyte composition change over time. The objective of this study is to gain insights into the influence of natural crossbreeding and parental lineage in the seed bacterial endophytic communities of two pure inbred lines exploring contributions of the two most important sources of plant endophytes - colonization from external sources and vertical transmission via seeds. Total genomic DNA was isolated from rice seeds and bacterial DNA was selectively amplified by PCR. The diversity of endophytic bacteria was studied through Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Diversity between the original parents and the pure inbred line may show significant differences in terms of richness, evenness and diversity indices. Heat maps reveal astonishing contributions of both or either parents (IR29 ${\times}$ Pokkali and AT401 ${\times}$ IR31868) in the shaping of the bacterial seed endophytes of the hybrid, FL478 and IC32, respectively. Most of the T-RFs of the subsequent pure inbred line could be traced to any or both of the parents. Comparison of common and genotype-specific T-RFs of parents and their offspring reveals that majority of the T-RFs are shared suggesting higher transmission of bacterial communities common to both parents. The parents influence the bacterial community of their offspring. Unique T-RFs of the offspring also suggest external sources of colonization particularly as the seeds are cultivated in different ecogeographical locations. This study showed that host parental lines contributed greatly in the shaping of bacterial seed endophytes of their offspring. It also revealed transmission and potential conservation of core seed bacterial endophytes that generally become the dominant microbiota in the succeeding generations of plant hosts.

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Induction of Defense Response Against Rhizoctonia solani in Cucumber Plants by Endophytic Bacterium Bacillus thuringiensis GS1

  • Seo, Dong-Jun;Nguyen, Dang-Minh-Chanh;Song, Yong-Su;Jung, Woo-Jin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.407-415
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    • 2012
  • An endophytic bacterium, Bacillus thuringiensis GS1, was isolated from bracken (Pteridium aquilinum) and found to have maximal production of chitinase (4.3 units/ml) at 5 days after culture. This study investigated the ability of B. thuringiensis GS1 to induce resistance to Rhizoctonia solani KACC 40111 (RS) in cucumber plants. Chitinase activity was greatest in RS-treated plants at 4 days. ${\beta}$-1,3-Glucanase activity was highest in GS1-treated plants at 5 days. Guaiacol peroxidase (GPOD) activity increased continuously in all treated plants for 5 days. Ascorbate peroxidase (APX) activity in RS-treated plants was increased 1.5-fold compared with the control at 4 days. Polyphenol oxidase (PPO) activity in RS-treated plants was increased 1.5-fold compared with the control at 3 days. At 5 days after treatment, activity staining revealed three bands with chitinase activity (Ch1, Ch2, and Ch3) on SDS-PAGE of cucumber plants treated with GS1+RS, whereas only one band was observed for RS-treated plants (Ch2). One GPOD isozyme (Gp1) was also observed in response to treatment with RS and GS1+RS at 4 days. One APX band (Ap2) was present on the native-PAGE gel of the control, and GS1- and GS1+RS-treated plants at 1 day. PPO bands (Po1 and Po2) from RS- and GS1+RS-treated plants were stronger than in the control and GS1-treated plants upon native-PAGE at 5 days. Taken together, these results indicate that the induction of PR proteins and defense-related enzymes by B. thuringiensis GS1 might have suppressed the damping-off caused by R. solani KACC 40111 in cucumber plants.

Multi-Function of a New Bioactive Secondary Metabolite Derived from Endophytic Fungus Colletotrichum acutatum of Angelica sinensis

  • Ramy S. Yehia
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.806-822
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    • 2023
  • In the current study we assessed a new crystallized compound, 5-(1-hydroxybutyl)-4-methoxy-3-methyl-2H-pyran-2-one (C-HMMP), from the endophytic fungus Colletotrichum acutatum residing in the medicinal plant Angelica sinensis for its in vitro antimicrobial, antibiofilm, antioxidant, antimalarial, and anti-proliferative properties. The promising compound was identified as C-HMMP through antimicrobial-guided fraction. The structure of C-HMMP was unambiguously confirmed by 2D NMR and HIRS spectroscopic analysis. Antimicrobial property testing of C-HMMP showed it to be effective against a variety of pathogenic bacteria and fungi with MICs ranging from 3.9 to 31.25 ㎍/ml. The compound displayed excellent antibiofilm activity against C. albicans, S. aureus, and K. pneumonia. Furthermore, the antimalarial and radical scavenging activities of C-HMMP were clearly dosedependent, with IC50 values of 0.15 and 131.2 ㎍/ml. The anti-proliferative activity of C-HMMP against the HepG-2, HeLa, and MCF-7 cell lines in vitro was investigated by MTT assay, revealing notable anti-proliferative activity with IC50 values of 114.1, 90, and 133.6 ㎍/ml, respectively. Moreover, CHMMP successfully targets topoisomerase I and demonstrated beneficial anti-mutagenicity in the Ames test against the reactive carcinogenic mutagen, 2-aminofluorene (2-AF). Finally, the compound inhibited the activity of α-glucosidase and α-amylase with IC50 values of 144.7 and 118.6 ㎍/ml, respectively. To the best of our knowledge, the identified compound C-HMMP was obtained for the first time from C. acutatum of A. sinensis, and this study demonstrated that C-HMMP has relevant biological significance and could provide better therapeutic targets against disease.

Prospecting endophytic colonization in Waltheria indica for biosynthesis of silver nanoparticles and its antimicrobial activity

  • Nirmala, C.;Sridevi, M.
    • Advances in nano research
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    • v.13 no.4
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    • pp.325-339
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    • 2022
  • Endophytes ascertain a symbiotic relationship with plants as promoters of growth, defense mechanism etc. This study is a first report to screen the endophytic population in Waltheria indica, a tropical medicinal plant. 5 bacterial and 3 fungal strains in leaves, 3 bacterial and 1 yeast species in stems were differentiated morphologically and identified by biochemical and molecular methods. The phylogenetic tree of the isolated endophytes was constructed using MEGA X. Silver nanoparticles were biosynthesized from a rare endophytic bacterium Cupriavidus metallidurans isolated from the leaf of W. indica. The formation of silver nanoparticles was confirmed by UV-Visible spectrophotometer that evidenced a strong absorption band at 408.5 nm of UV-Visible range with crystalline nature and average particle size of 16.4 nm by Particle size analyzer. The Fourier Transform Infra-Red spectrum displayed the presence of various functional groups that stabilized the nanoparticles. X-ray diffraction peaks were conferred to face centered cubic structure. Transmission Electron Microscope and Scanning Electron Microscope revealed the spherical-shaped, polycrystalline nature with the presence of elemental silver analyzed by Energy Dispersive of X-Ray spectrum. Selected area electron diffraction also confirmed the orientation of AgNPs at 111, 200, 220, 311 planes similar to X-ray diffraction analysis. The synthesized nanoparticles are evaluated for antimicrobial activity against 7 bacterial and 3 fungal pathogens. A good zone of inhibition was observed against pathogenic bacteria than fungal pathogens. Thus the study could hold a key aspect in drug discovery research and other pharmacological conducts of human clinical conditions.

Classification of Bacillus Beneficial Substances Related to Plants, Humans and Animals

  • Mongkolthanaruk, Wiyada
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1597-1604
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    • 2012
  • Genus Bacillus is a spore-forming bacterium that has unique properties in cell differentiation, allowing the forming of spores in stress conditions and activated in the vegetative cell, with suitable environments occurring during the life cycle acting as a trigger. Their habitat is mainly in soil; thus, many species of Bacillus are associated with plants as well as rhizosphere bacteria and endophytic bacteria. Signal transduction is the principal mechanism of interactions, both within the cell community and with the external environment, which provides the subsequent functions or properties for the cell. The antimicrobial compounds of Bacillus sp. are potentially useful products, which have been used in agriculture for the inhibition of phytopathogens, for the stimulation of plant growth, and in the food industry as probiotics. There are two systems for the synthesis of these substances: nonribosomal synthesis of cyclic lipopeptides (NRPS) and polyketides (PKS). For each group, the structures, properties, and genes of the main products are described. The different compounds described and the way in which they co-exist exhibit the relationship of Bacillus substances to plants, humans, and animals.

Isolation and characterization of an antifungal substance from Burkholderia cepacia, an endophytic bacteria obtained from roots of cucumber.

  • Park, J.H.;Park, G.J.;Lee, S.W;Jang, K.S.;Park, Y.H.;Chung, Y.R.;Cho, K.Y.;Kim, J.C.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.95.2-96
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    • 2003
  • In order to develop a new microbial fungicide for the control of vegetable diseases using endophytic bacteria, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth media, their antifungal activities were screened by in vivo bioassays against Botrytis cinerea(tomato gray mold), Pythium ultimum(cucumber damping-off), Phytopkhora infestans(tomato late blight), Colletotrichum orbiculare(cucumber anthracnose), and Blumeria graminis f. sp. hordei(barley powdery mildew). As the results of screening, 38 bacterial strains showed potent antifungal activities against at least one of 5 plant pathogens. A bacterial strain EB072 displayed potent disease control activities against 3 plant diseases. Among the bacterial strains with a potent antifungal activity against cucunlber anthracnose, three bacterial strains, EB054, EB151 and EB215, also displayed a potent in vitro antifungal activity against C. acutatum, a fungal agent causing pepper anthracnose. A bacterial strain EB215 obtained from roots of cucumber was identified as Burkholderia cepacia based on its physiological and biochemical characteristics and 165 rRNA gene sequence. An antifungal substance was isolated from the liquid cultures of B. cepacia EB215 strain by ethyl acetate partitioning, repeated silica gel column chromatography, and invitro bioassay, Its structural determination is in progress by various instrumental analyses.

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