• Title/Summary/Keyword: root isolates

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Diversity and Bioactive Potential of Culturable Fungal Endophytes of Medicinal Shrub Berberis aristata DC.: A First Report

  • Sharma, Supriya;Gupta, Suruchi;Dhar, Manoj K.;Kaul, Sanjana
    • Mycobiology
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    • v.46 no.4
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    • pp.370-381
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    • 2018
  • Bioactive natural compounds, isolated from fungal endophytes, play a promising role in the search for novel drugs. They are an inspiring source for researchers due to their enormous structural diversity and complexity. During the present study fungal endophytes were isolated from a well-known medicinal shrub, Berberis aristata DC. and were explored for their antagonistic and antioxidant potential. B. aristata, an important medicinal shrub with remarkable pharmacological properties, is native to Northern Himalayan region. A total of 131 endophytic fungal isolates belonging to eighteen species and nine genera were obtained from three hundred and thirty surface sterilized segments of different tissues of B. aristata. The isolated fungi were classified on the basis of morphological and molecular analysis. Diversity and species richness was found to be higher in leaf tissues as compared to root and stem. Antibacterial activity demonstrated that the crude ethyl acetate extract of 80% isolates exhibited significant results against one or more bacterial pathogens. Ethyl acetate extract of Alternaria macrospora was found to have potential antibacterial activity. Significant antioxidant activity was also found in crude ethyl acetate extracts of Alternaria alternata and Aspergillus flavus. Similarly, antagonistic activity of the fungal endophytes revealed that all antagonists possessed inhibition potential against more than one fungal pathogen. This study is an important step towards tapping endophytic fungal diversity for bioactive metabolites which could be a step forward towards development of novel therapeutic agents.

Analysis of Microbial Community Structure in Soil and Crop Root System II. Analysis of soil microbial community structure in different soil Environmental conditions by MIDI and DNA analyses (토양과 작물근계의 미생물군집 구조 해석 II. MIDI 및 DNA 분석에 의한 토양환경별 미생물 군집 해석)

  • Ryu, Jin-Chang;Kwon, Soon-Wo;Kim, Jong-Shik;Suh, Jang-Sun;Jung, Beung-Gan;Choi, Sun-Shik
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

Biological control efficacy on Sclerotinia rot(Sclerotinia sclerotiorum) by the use of antifungal agent some Bacillus sp.

  • Hong, Yeon-Kyu;Lee, Bong-Choon;Shin, Dong-Beom;Hyun, Jong-Nae;Kang, Hang-Won;Park, Sung-Tae
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.106-107
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    • 2003
  • The effect of biological control agent Bacillus sp. (BAC03-3-1, BAC03-3-2, BAC02-4) on pre- and postemergence Sclerotinia rot of perilla (Perilla frutescens var. japonica) caused by Sclerotinia sclerotiorum was determined from greenhouse field trials. The ability of this antagonist to reduce germination of sclerotia of S. sclerotiorum was also evaluated. In the greenhouse, suspension of BAC03-3-1 application as root drench of perilla, which provided as little as 10$\^$7/ cells/ $m\ell$ per gram of soil, significantly increased plant stand in pathogen-infested soil over that in the untreated control. All three isolates reduced the germination of sclerotia of S. sclerotiorum in loamy sand soils in the greenhouse. In loamy sand amended with rice bran the sclerotial germination was inversely correlated (r = -0.79) with perilla stand in the greenhouse. However, a higher rate of bacterial suspension with rice bran(Ig dwt./100g soil) than that applied with bacterial suspensions only was necessary to achieve a comparable reduction in sclerotial germination. In field study, all three isolates added to soil to provide 10$\^$7/ cells/$m\ell$ per gram significantly prevented Sclerotinia rot (73-85%) after 35 days of growth. The isolate BAC02-4, BAC03-3-1 and BAC03-3-2 gave final stands of 65 to 75, 60 to 70, and 55 to 60%, respectively. The addition of rice bran(1 %) to loamy sand in the field resulted in a 10-fold increase in propagule numbers of the three isolates within 10 days of application.

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Isolation and Identification of Endophytic Bacteria with Plant Growth Promoting Activity and Biocontrol Potential from Wild Pistachio Trees

  • Etminani, Faegheh;Harighi, Behrouz
    • The Plant Pathology Journal
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    • v.34 no.3
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    • pp.208-217
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    • 2018
  • In this study, samples were collected from the leaves and stems of healthy wild Pistachio trees (Pistacia atlantica L.) from various locations of Baneh and Marivan regions, Iran. In total, 61 endophytic bacteria were isolated and grouped according to phenotypic properties. Ten selected isolates from each group were further identified by partial sequencing of the 16S rRNA gene. Based on the results, isolates were identified as bacteria belonging to Pseudomonas, Stenotrophomonas, Bacillus, Pantoea and Serratia genus. The ability of these isolates was evaluated to phytohormone production such as auxin and gibberellin, siderophore production, phosphate solubilization, atmospheric nitrogen fixation, protease and hydrogen cyanide production. All strains were able to produce the plant growth hormone auxin and gibberellin in different amounts. The majority of strains were able to solubilize phosphate. The results of atmospheric nitrogen fixation ability, protease and siderophore production were varied among strains. Only Ba66 could produce a low amount of hydrogen cyanide. The results of biocontrol assay showed that Pb78 and Sp15 strains had the highest and lowest inhibition effects on bacterial plant pathogens, Pseudomonas syringae pv. syringae Pss20 and Pseudomonas tolaasii Pt18 under in vitro condition. Pb3, Pb24 and Pb71 strains significantly promote root formation on carrot slices. To our knowledge this is the first report of the isolation of endophytic bacterial strains belonging to Pantoea, Bacillus, Pseudomonas, Serratia and Stenotrophomonas genus from wild pistachio trees with plant growth promoting potential and biocontrol activity.

Halotolerant Spore-Forming Gram-Positive Bacterial Diversity Associated with Blutaparon portulacoides (St. Hill.) Mears, a Pioneer Species in Brazilian Coastal Dunes

  • Barbosa Deyvison Clacino;Irene Von Der Weid;Vaisman Natalie;Seldin Lucy
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.193-199
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    • 2006
  • Halotolerant spore-forming Gram-positive bacteria were isolated from the root, rhizosphere, and non-rhizosphere soil of Blutaparon portulacoides. The different isolates were characterized genetically using an amplified ribosomal DNA restriction analysis (ARDRA), and phenotypically based on their colonial morphology, physiology, and nutritional requirements. Three different 16S rRNA gene-based genotypes were observed at a 100% similarity using the enzymes HinfI, MspI, and RsaI, and the phenotypic results also followed the ARDRA groupings. Selected strains, representing the different ARDRA groups, were analyzed by 16S rDNA sequencing, and members of the genera Halobaeillus, Virgibacillus, and Oceanobacillus were found. Two isolates showed low 16S rDNA sequence similarities with the closest related species of Halobacillus, indicating the presence of new species among the isolates. The majority of the strains isolated in this study seemed to belong to the species O. iheyensis and were compared using an AP-PCR to determine whether they had a clonal origin or not. Different patterns allowed the grouping of the strains according to Pearson's coefficient, and the resulting dendrogram revealed the formation of two main clusters, denoted as A and B. All the strains isolated from the soil were grouped into cluster A, whereas cluster B was exclusively composed of the strains associated with the B. portulacoides roots. This is the first report on the isolation and characterization of halotolerant spore-forming Gram-positive bacteria that coexist with B. portulacoides. As such, these new strains may be a potential source for the discovery of bioactive compounds with industrial value.

Assessment of Root-Associated Paenibacillus polymyxa Groups on Growth Promotion and Induced Systemic Resistance in Pepper

  • Phi, Quyet-Tien;Park, Yu-Mi;Seul, Keyung-Jo;Ryu, Choong-Min;Park, Seung-Hwan;Kim, Jong-Guk;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1605-1613
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    • 2010
  • Twenty-nine P. polymyxa strains isolated from rhizospheres of various crops were clustered into five genotypic groups on the basis of BOX-PCR analysis. The characteristics of several plant growth-promoting factors among the isolates revealed the distinct attributes in each allocated group. Under gnotobiotic conditions, inoculation of pepper roots with P. polymyxa isolates significantly increased the biomass in 17 of total 29 treated plants with untreated plants. Experiments on induced systemic resistance (ISR) against bacterial spot pathogen Xanthomonas axonopodis pv. vesicatoria in pepper by P. polymyxa strains were conducted and only one isolate (KNUC265) was selected. Further studies into ISR mediation by the KNUC265 strain against the soft-rot pathogen Erwinia carotovora subsp. carotovora in tobacco demonstrated that the tobacco seedlings exposed to either bacterial volatiles or diffusible metabolites exhibited a reduction in disease severity. In conclusion, ISR and plant growth promotion triggered by P. polymyxa isolates were systemically investigated on pepper for the first time. The P. polymyxa KNUC265 strain, which elicited both ISR and plant growth promotion, could be potentially used in improving the yield of pepper and possibly of other crops.

Diversity of Fungal Endophytes in Various Tissues of Panax ginseng Meyer Cultivated in Korea

  • Park, Young-Hwan;Lee, Soon-Gu;Ahn, Doek-Jong;Kwon, Tae-Ryong;Park, Sang-Un;Lim, Hyoun-Sub;Bae, Han-Hong
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.211-217
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    • 2012
  • Endophytic fungi were isolated from various tissues (root, stem, petiole, leaf, and flower stalk) of 3- and 4-year-old ginseng plants (Panax ginseng Meyer) cultivated in Korea. The isolated endophytic fungi were identified based on the sequence analysis of the internal transcribed spacer (ITS), 1-5.8-ITS 2. A morphological characterization was also conducted using microscopic observations. According to the identification, 127 fungal isolates were assigned to 27 taxa. The genera of Phoma, Alternaria and Colletotrichum were the most frequent isolates, followed by Fusarium, Entrophospora and Xylaria. Although 19 of the 27 taxa were identified at the species level, the remainder were classified at the genus level (6 isolates), phylum level (Ascomycota, 1 isolate), and unknown fungal species (1 isolate). Endophytic fungi of 13 and 19 species were isolated from 3- and 4-year-old ginseng plants, respectively, and Phoma radicina and Fusarium solani were the most frequently isolated species colonizing the tissues of the 3- and 4-year-old ginseng plants, respectively. The colonization frequency (CF%) was dependant on the age and tissue examined: the CFs of the roots and stems in the 3-year-old ginseng were higher than the CF of tissues in the 4-year-old plants. In contrast, higher CFs were observed in the leaves and petioles of 4-year-old plants, and endophytic fungi in the flower stalks were only detected in the 4-year-old plants. In conclusion, we detected diverse endophytic fungi in ginseng plants, which were distributed differently depending on the age and tissue examined.

Isolations of Orchid Mycorrhizal Fungi from the Korean Native Orchid Plants (한국의 자생 난과식물에서 난균근균(蘭菌根菌)(Orchid Mycorrhizal Fungi) 분리)

  • Lee, Sang-Sun;Riew, Hee-Kyun;Paek, Kee-Yoeup
    • The Korean Journal of Mycology
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    • v.25 no.2 s.81
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    • pp.101-110
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    • 1997
  • This study was to identify the orchid mycorrhizal fungi and to test whether the orchid plants antificially inoculated with this fungus showed better growth them uninoculated plants. Symbioses in the root cells of the native plants of Cymbidium goeringii collected were observed and the digestive forms of peletons were also observed in various native roots. Two types of hyphae, thick $(7{\sim}10\;{\mu}m)$ and thin $(2{\sim}4\;{\mu}m)$ in thickness, were conclusively found to be from various native orchid roots. The symbiotic fungus was isolated by several agars and identified as a Rhizoctonia repens or a R. endophytica var. endophytica. Symbioses on the plantlets of C. karnan and Cymbidium hybrid 'Onomoron' were evaluated as the isolates inoculated on oatmeal agars. The growth of plantlets were measured with the formations of mycorrhizae in the roots. R. repens was shown to be the better isolate than the other in growth stimulation of plantlets on oatmeal agars when grown for two months. The two types of hyphae in the root cells under nature were speculated from the different fungal isolates of Rhizoctonia. Further isolates would be needed for application works for the orchid industries.

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'Kresek' Disease in Korea. I The Grouping of the Pathogens and Reproduction of 'Kresek' (한국에 있어서의 'Kresek'에 관한 연구 I 'Kresek' 발생지의 병원균 균형 및 병징 재현에 관한 시험)

  • Choi Yong Chull;Cho Eng Haeng;Chung Bong Jo;Cho Yong Sup;Yoo Yeon Hean
    • Korean journal of applied entomology
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    • v.16 no.1 s.30
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    • pp.1-6
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    • 1977
  • The 'Kresek' disease in Korea caused by Xanthomonas oryzae was first found from eight counties in Jeon-nam and Kyungnam province in 1976. The study has been carried mainly on the grouping of pathogens and reproduction of the symptoms on Milyang #23, the variety had shown severe damages at the fields, by using the isolates from Hwa-sun and Kwang-san counties where the first epidemics took place. 1. The 'Kresek' disease was found mainly on Milyang #23, a new variety, at Hwa-sun, Mu-an, Kang-jin. Yung-am, Gog-sung counties in Jeon-nam province and Jin-yang county in Kyung-nam province. 2. The groups of 'Kresek' causing pathogens were the same of those producing bacterial leaf blight symptoms such as group I , II, IN and V of Xanthomonas oryzae. 3. Seventeen out of 21 isolates from Hwa-sun county where the first and severe damage found belonged to group IV, 2 to group II and 2 to group V All of 5 isolates from Kwang-san county belonged to the group IN. 4. The 'Kresek' type symptom could reproduced within 5 days after inoculation to seedlings by using root tut, spray and needle inoculation methods. j. The most of effective method for the inoculation was root cut, and then were spray and needle method. respectively. The higher concentration of inoculum produced the higher disease incidence.

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Genetic Diversity of Korean Cylindrocarpon destructans Based on Virulence Aassay and RAPD Analysis (병원성 검정 및 RAPD 분석에 의한 국내 인삼뿌리썩음병균(Cylindrocarpon destructans)의 유전적 다양성)

  • Seo, Mun-Won;Kim, Sun-Ick;Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.16-21
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    • 2011
  • Ginseng root rot caused by Cylindrocarpon destructans is one of the most destructive diseases of ginseng(Panax ginseng). We analyzed the features of the species through pathogenicity test and genetic diversity analysis of C. destructans in Korea, for its application as basic data to attempt for effective control. C. destructans isolated from rotted ginseng roots exhibited a variety of colonial colors on media. It was assumed that there may exist genetic diversity in the population by the diversity of pathogenicity among isolates observed when artificially inoculated into ginseng roots. Pathogenicity tests using ex vivo wound inoculation with agar mixture inoculation on ginseng roots were performed similar results as were observed appear to be useful for rapid pathogen inspection. According to RAPD analysis results, Korean C. destructans isolates formed a single genetic group which can be distinguished readily from closely related other fungi. C. destructans group was divided into two small groups. Therefore, we were able to confirm pathogenicity and genetic difference between the isolates in each of the groups of the pathogen.