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Sample records for endophytic actinomycetes isolated

  1. Diversity, bioactivities, and metabolic potentials of endophytic actinomycetes isolated from traditional medicinal plants in Sichuan, China.

    PubMed

    Qiu, Peng; Feng, Zhi-Xiang; Tian, Jie-Wei; Lei, Zu-Chao; Wang, Lei; Zeng, Zhi-Gang; Chu, Yi-Wen; Tian, Yong-Qiang

    2015-12-01

    The present study was designed to determine the taxonomic diversity and metabolic activity of the actinomycetes community, including 13 traditional medicinal plants collected in Sichuan province, China, using multiple approaches such as morphological and molecular identification methods, bioactivity assays, and PCR screening for genes involved in antibiotics biosynthesis. 119 endophytic actinomycetes were recovered; 80 representative strains were chosen for 16S rRNA gene partial sequence analyses, with 66 of them being affiliated to genus Streptomyces and the remaining 14 strains being rare actinomycetes. Antimicrobial tests showed that 12 (15%) of the 80 endophytic actinomycetes displayed inhibitory effects against at least one indicator pathogens, which were all assigned to the genus Streptomyces. In addition, 87.5% and 58.8% of the isolates showed anticancer and anti-diabetic activities, respectively. Meanwhile, the anticancer activities of the isolates negatively correlated with their anti-diabetic activities. Based on the results of PCR screening, five genes, PKS-I, PKS-II, NRPS, ANSA, and oxyB, were detected in 55.0%, 58.8%, 90.0%, 18.8% and 8.8% of the 80 actinomycetes, respectively. In conclusion, the PCR screening method employed in the present study was conducive for screening and selection of potential actinomycetes and predicting potential secondary metabolites, which could overcome the limitations of traditional activity screening models.

  2. Antibacterial Activity of Endophytic Actinomycetes Isolated from the Medicinal Plant Vochysia divergens (Pantanal, Brazil).

    PubMed

    Gos, Francielly M W R; Savi, Daiani C; Shaaban, Khaled A; Thorson, Jon S; Aluizio, Rodrigo; Possiede, Yvelise M; Rohr, Jürgen; Glienke, Chirlei

    2017-01-01

    Endophytic actinomycetes from medicinal plants produce a wide diversity of secondary metabolites (SM). However, to date, the knowledge about endophytes from Brazil remains scarce. Thus, we analyzed the antimicrobial potential of 10 actinomycetes isolated from the medicinal plant Vochysia divergens located in the Pantanal sul-mato-grossense, an unexplored wetland in Brazil. Strains were classified as belonging to the Aeromicrobium, Actinomadura, Microbacterium, Microbispora, Micrococcus, Sphaerisporangium, Streptomyces, and Williamsia genera, through morphological and 16S rRNA phylogenetic analyzes. A susceptibility analysis demonstrated that the strains were largely resistant to the antibiotics oxacillin and nalidixic acid. Additionally, different culture media (SG and R5A), and temperatures (28 and 36°C) were evaluated to select the best culture conditions to produce the active SM. All conditions were analyzed for active metabolites, and the best antibacterial activity was observed from metabolites produced with SG medium at 36°C. The LGMB491 (close related to Aeromicrobium ponti) extract showed the highest activity against methicillin-resistant Staphylococcus aureus (MRSA), with a MIC of 0.04 mg/mL, and it was selected for SM identification. Strain LGMB491 produced 1-acetyl-β-carboline (1), indole-3-carbaldehyde (2), 3-(hydroxyacetyl)-indole (4), brevianamide F (5), and cyclo-(L-Pro-L-Phe) (6) as major compounds with antibacterial activity. In this study, we add to the knowledge about the endophytic community from the medicinal plant V. divergens and report the isolation of rare actinomycetes that produce highly active metabolites.

  3. Biotechnological potential of endophytic actinomycetes associated with Asteraceae plants: isolation, biodiversity and bioactivities.

    PubMed

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida

    2014-04-01

    Endophytic actinomycetes from five Asteraceae plants were isolated and evaluated for their bioactivities. From Parthenium hysterophorus, Ageratum conyzoides, Sonchus oleraceus, Sonchus asper and Hieracium canadense, 42, 45, 90, 3, and 2 isolates, respectively, were obtained. Of the isolates, 86 (47.2 %) showed antimicrobial activity. Majority of the isolates were recovered from the roots (n = 127, 69.7 %). The dominant genus was Streptomyces (n = 96, 52.7 %), while Amycolatopsis, Pseudonocardia, Nocardia and Micromonospora were also recovered. Overall, 36 of the 86 isolates were significantly bioactivity while 18 (20.9 %) showed strong bioactivity. In total, 52.1 and 66.6 % showed potent cytotoxicity and antioxidant activities. The LC50 for 15 strains was <20 μg/ml. Compared to the ascorbate standard (EC50 0.34 μg/ml), all isolates gave impressive results with notable EC50 values of 0.65, 0.67, 0.74 and 0.79 μg/ml.

  4. Nonomuraea stahlianthi sp. nov., an endophytic actinomycete isolated from the stem of Stahlianthus campanulatus.

    PubMed

    Niemhom, Nantawan; Chutrakul, Chanikul; Suriyachadkun, Chanwit; Thawai, Chitti

    2017-08-01

    A novel endophytic actinomycete, designated strain SC1-1T, was isolated from sterilized stem tissue from Stahlianthus campanulatus collected in Udon Thani province, Thailand. The isolate formed short chains of spores on aerial mycelium and presented meso-diaminopimelic acid in the cell wall peptidoglycan. Glucose, madurose, mannose, rhamnose and ribose were observed as sugars in the cells. The cell membrane phospholipids were phosphatidylethanolamine, phosphatidylmethylethanolamine, hydroxy-phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and ninhydrin-positive phosphoglycolipids. The major menaquinones were MK-9(H4) and MK-9(H2). The main cellular fatty acids were iso-C16:0, 10-methyl C17 : 0 and C17 : 1ω6c. A high G+C content (70.7 mol%) was present in the genomic DNA. The taxonomic position based on the 16S rRNA gene sequence analysis revealed that strain SC1-1T belonged to the genus Nonomuraea and shared the highest 16S rRNA gene sequence similarity value with Nonomuraea dietziae DSM 44320T (98.82 %), followed by Nonomuraea africana IFO 14745T (98.58 %), Nonomuraea jabiensis A4036T (98.43 %), Nonomuraea endophytica YIM 65601T (98.36 %), Nonomuraea purpurea 1SM4-01T (98.34 %), Nonomuraea angiospora IFO 13155T (98.29 %), Nonomuraea roseola IFO 14685T (98.23 %) and Nonomuraea recticatena IFO 14525T (98.23 %). On the basis of the DNA-DNA hybridization relatedness and including the physiological and biochemical characteristics, strain SC1-1T should be judged as a novel species of the genus Nonomuraea, for which the name Nonomuraea stahlianthi sp. nov. is proposed. The type strain is strain SC1-1T (=BCC 66361T=NBRC 110006T).

  5. Phytohabitans kaempferiae sp. nov., an endophytic actinomycete isolated from the leaf of Kaempferia larsenii.

    PubMed

    Niemhom, Nantawan; Chutrakul, Chanikul; Suriyachadkun, Chanwit; Thawai, Chitti

    2016-08-01

    A novel endophytic actinomycete, designated strain KK1-3T, which formed single spores and long chains of spores (more than 10 spores) was isolated from surface-sterilized Kaempferia larsenii leaf collected from Ubon Ratchathani province, Thailand. The isolate contained l-lysine, meso-diaminopimelic acid and hydroxyl diaminopimelic acid in the cell-wall peptidoglycan. The whole-cell sugars included glucose, mannose, rhamnose, ribose, galactose and xylose. The characteristic phospholipids were phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and phosphoglycolipids. The predominant menaquinones were MK-10(H8), MK-10(H6) and MK-10(H4). The predominant cellular fatty acids were anteiso-C17 : 0 and iso-C16 : 0. The G+C content of the genomic DNA was 71 mol%. Phylogenetic analysis using 16S rRNA gene sequences revealed that strain KK1-3T should be classified as representing a member of the genus Phytohabitans. The similarity values of sequences between this strain and those of the closely related species, Phytohabitans houttuyneae K11-0057T (99.0 %), Phytohabitans suffuscus K07-0523T (98.9 %), Phytohabitans flavus K09-0627T (98.6 %) and Phytohabitans rumicisK11-0047T (98.1 %) were observed. The DNA-DNA hybridization result and some physiological and biochemical properties indicated that KK1-3T could be readily distinguished from its closest phylogenetic relatives. On the basis of these phenotypic and genotypic data, this strain represents a novel species, for which the name Phytohabitans kaempferiae sp. nov. is proposed. The type strain is strain KK1-3T (=BCC 66360T =NBRC 110005T).

  6. Actinoallomurus bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Wang, Haiyan; Jin, Pinjiao; Zheng, Weijia; Chu, Liyang; Liu, Chongxi; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing

    2015-08-01

    A novel endophytic actinomycete, strain NEAU-TX1-15(T), was isolated from moss, collected from Wuchang, Heilongjiang province, north China. A polyphasic taxonomic study was carried out to establish the status of strain NEAU-TX1-15(T). Morphological and chemotaxonomic properties of strain NEAU-TX1-15(T) are consistent with the description of the genus Actinoallomurus. Strain NEAU-TX1-15(T) was observed to form short spiral or looped spore chains on aerial hyphae. The cell wall peptidoglycan was found to contain lysine and meso-diaminopimelic acid. The major menaquinones were identified as MK-9(H6) and MK-9(H8). The only phospholipid identified was phosphatidylglycerol. The major fatty acid was identified as iso-C16:0. Analysis of the 16S rRNA gene sequence supports the assignment of the novel strain to the genus Actinoallomurus, as it exhibits 99.2 % gene sequence similarity to that of Actinoallomurus yoronensis NBRC 103686(T). However, the low level of DNA-DNA relatedness allowed the strain to be differentiated from its close relative. Moreover, strain NEAU-TX1-15(T) could also be differentiated from A. yoronensis NBRC 103686(T) and other Actinoallomurus species showing high 16S rRNA gene sequence similarity (>98.0 %) by cultural and physiological characteristics. Therefore, the combination of phenotypic and chemotaxonomic data, and the DNA-DNA hybridization value, indicated that strain NEAU-TX1-15(T) represents a novel species of the genus Actinoallomurus for which the name Actinoallomurus bryophytorum sp. nov. is proposed. The type strain is NEAU-TX1-15(T) (=CGMCC 4.7200(T) = JCM 30340(T)).

  7. Streptomyces bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Jin, Pinjiao; Liu, Chongxi; Ma, Zhaoxu; Zhao, Junwei; Li, Jiansong; Wang, Xiangjing; Xiang, Wensheng

    2016-09-01

    A novel endophytic actinomycete, designated strain NEAU-HZ10(T) was isolated from moss and characterised using a polyphasic approach. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Strain NEAU-HZ10(T) formed grayish aerial mycelia, which differentiated into straight to flexuous chains of cylindrical spores. The cell wall peptidoglycan was found to contain LL-diaminopimelic acid. Predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The polar lipid profile was found to consist of phosphatidylethanolamine, phosphatidylinositol and two unidentified phospholipids. The major fatty acids were identified as iso-C16:0, anteiso-C15:0 and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-HZ10(T) belongs to the genus Streptomyces and exhibits high sequence similarity to Streptomyces cocklensis DSM 42063(T) (98.9 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-HZ10(T) clustered with S. cocklensis DSM 42063(T), Streptomyces yeochonensis CGMCC 4.1882(T) (98.7 %), Streptomyces paucisporeus CGMCC 4.2025(T) (98.4 %) and Streptomyces yanglinensis CGMCC 4.2023(T) (98.1 %). However, a combination of DNA-DNA hybridisation results and some phenotypic characteristics indicated that strain NEAU-HZ10(T) can be distinguished from its phylogenetically closely related strains. Therefore, it is proposed that strain NEAU-HZ10(T) represents a novel species of the genus Streptomyces for which the name Streptomyces bryophytorum sp. nov. is proposed. The type strain is NEAU-HZ10(T) (= CGMCC 4.7151(T) = DSM 42138(T)).

  8. Asanoa endophytica sp. nov., an endophytic actinomycete isolated from the rhizome of Boesenbergia rotunda.

    PubMed

    Niemhom, Nantawan; Chutrakul, Chanikul; Suriyachadkun, Chanwit; Thawai, Chitti

    2016-01-12

    A novel Gram-stain-positive, non motile endophytic actinomycete, designated strain BR3-1T, which produced spore chains borne on the tip of short sporophores was isolated from the rhizome of Boesenbergia rotunda collected from Udon Thani province, Thailand. This strain was investigated for its taxonomic position using polyphasic approach. The strain contained 3-hydroxy-diaminopimelic acid and meso-diaminopimelic acid in the cell-wall peptidoglycan. The whole-cell sugars contained glucose, mannose, rhamnose, ribose and xylose. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides were found as the characteristic phospholipids. The predominant menaquinones were MK-10(H8) and MK-10(H6). The major cellular fatty acids were iso-C16:0, iso-C15:0 and anteiso-C15:0. The G+C content of the genomic DNA was 71.4 mol%. On the basis of 16S rRNA gene sequences analysis revealed that strain BR3-1T belonged to the genus Asanoa and was most closely related to Asanoa ishikariensis (99.39%), Asanoa iriomotensis (99.31%), Asanoa siamensis (99.17%), Asanoa ferruginea (98.84%) and Asanoa hainanensis (98.71%). The DNA-DNA relatedness value between the strain BR3-1T and its phylogenetically closest relatives were in the range of 15.4% ± 1.2 to 45.8% ± 2.6. In addition, some physiological and biochemical properties indicated that strain BR3-1T could be readily distinguished from all type strains in the genus Asanoa. Thus, strain BR3-1T should be represented as a novel species, for which the name Asanoa endophytica sp. nov. is proposed. The type strain is BR3-1T (=BCC 66355T =NBRC 110002T).

  9. Isolation, abundance and phylogenetic affiliation of endophytic actinomycetes associated with medicinal plants and screening for their in vitro antimicrobial biosynthetic potential

    PubMed Central

    Passari, Ajit K.; Mishra, Vineet K.; Saikia, Ratul; Gupta, Vijai K.; Singh, Bhim P.

    2015-01-01

    Microorganisms associated with medicinal plants are of interest as the producers of important bioactive compounds. To date, the diversity of culturable endophytic actinomycetes associated with medicinal plants is in its initial phase of exploration. In this study, 42 endophytic actinomycetes were isolated from different organs of seven selected medicinal plants. The highest number of isolates (n = 22, 52.3%) of actinomycetes was isolated from roots, followed by stems (n = 9, 21.4%), leaves (n = 6, 14.2%), flowers (n = 3, 7.1%), and petioles (n = 2, 4.7%). The genus Streptomyces was the most dominant among the isolates (66.6%) in both the locations (Dampa TRF and Phawngpuii NP, Mizoram, India). From a total of 42 isolates, 22 isolates were selected for further studies based on their ability to inhibit one of the tested human bacterial or fungal pathogen. Selected isolates were identified based on 16S rRNA gene analysis and subsequently the isolates were grouped to four different genera; Streptomyces, Brevibacterium, Microbacterium, and Leifsonia. Antibiotic sensitivity assay was performed to understand the responsible antimicrobials present in the isolates showing the antimicrobial activities and revealed that the isolates were mostly resistant to penicillin G and ampicillin. Further, antimicrobial properties and antibiotic sensitivity assay in combination with the results of amplification of biosynthetic genes polyketide synthase (PKS-I) and non-ribosomal peptide synthetase (NRPS) showed that the endophytic actinomycetes associated with the selected medicinal plants have broad-spectrum antimicrobial activity. This is the first report of the isolation of Brevibacterium sp., Microbacterium sp., and Leifsonia xyli from endophytic environments of medicinal plants, Mirabilis jalapa and Clerodendrum colebrookianum. Our results emphasize that endophytic actinomycetes associated with medicinal plants are an unexplored resource for the discovery of biologically active

  10. Diversity and bioprospecting of actinomycete endophytes from the medicinal plants.

    PubMed

    Nalini, M S; Prakash, H S

    2017-04-01

    The endophytic actinomycetes constitute one of the fascinating group of microorganisms associated with a wide range of plant species. The diversity of actinomycetes in plants and their tissue parts is a matter of debate as no consensus are derived between individual studies. Nevertheless, their diversity correlates with the occurrence in plant species harboured in unique regions of biologically diverse areas called "hot spots." Recent advances in the isolation techniques have facilitated the isolation of rare taxa from these environments. The biosynthetic ability of the endophytic actinomycetes has proven beyond doubt that these organisms have the potential to synthesize an array of compounds with novelty in structure and bioactivity and as a result are preferred in the natural product screening programs. In the years to come, the scientific world may await to discover many more novel actinomycete taxa with metabolic diversity and applications in therapeutics.

  11. Glycomyces endophyticus sp. nov., an endophytic actinomycete isolated from the root of Carex baccans Nees.

    PubMed

    Qin, Sheng; Wang, Hai-Bin; Chen, Hua-Hong; Zhang, Yu-Qin; Jiang, Cheng-Lin; Xu, Li-Hua; Li, Wen-Jun

    2008-11-01

    An actinomycete, designated strain YIM 56134(T), was isolated from the root of a Chinese medicinal plant, Carex baccans Nees, collected from Yunnan, south-west China, and subjected to a polyphasic taxonomic study. An analysis of 16S rRNA gene sequence similarities showed that strain YIM 56134(T) was a member of the genus Glycomyces, being most closely related to Glycomyces algeriensis NRRL B-16327(T) (99.0 % similarity), Glycomyces lechevalierae DSM 44724(T) (99.0 %), Glycomyces rutgersensis IFO 14488(T) (98.9 %) and Glycomyces harbinensis IFO 14487(T) (98.7 %). Strain YIM 56134(T) could be distinguished from other established Glycomyces species on the basis of relatively low sequence similarity (<97 %). Phenotypic and chemotaxonomic data supported the affiliation of this strain to the genus Glycomyces. The results of DNA-DNA hybridization and some physiological and biochemical tests allowed differentiation of the strain from related Glycomyces species. Therefore, strain YIM 56134(T) represents a novel species of the genus Glycomyces, for which the name Glycomyces endophyticus sp. nov. is proposed. The type strain is YIM 56134(T) (=KCTC 19152(T) =DSM 45002(T)).

  12. Plantactinosporasoyae sp. nov., an endophytic actinomycete isolated from soybean root [Glycine max (L.) Merr].

    PubMed

    Guo, Xiaowei; Guan, Xuejiao; Liu, Chongxi; Jia, Feiyu; Li, Jiansong; Li, Jinmeng; Jin, Pinjiao; Li, Wenchao; Wang, Xiangjing; Xiang, Wensheng

    2016-07-01

    A novel actinomycete, designated strain NEAU-gxj3T, was isolated from soybean root [Glycine max (L.) Merr.] collected from Harbin, Heilongjiang Province, China, and characterized using a polyphasic approach. The 16S rRNA gene sequence of strain NEAU-gxj3T showed highest similarity to those of Micromonospora equina Y22T (98.2 %) and Plantactinospora endophytica YIM 68255T (98.0 %). Phylogenetic analysis based on the 16S rRNA gene and gyrB gene demonstrated that the isolate clustered with the members of the genus Plantactinospora. The chemotaxonomic properties of strain NEAU-gxj3Twere also consistent with those of members of the genus Plantactinospora. The cell wall contained meso-diaminopimelic acid and whole-cell sugars were xylose, glucose and galactose. The predominant menaquinones were MK-10(H6), MK-9(H8), MK-10(H2) and MK-10(H4). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were identified as anteiso-C17 : 0, iso-C16 : 0, iso-C15 : 0 and C15 : 0. A combination of DNA-DNA hybridization result and some phenotypic characteristics indicated that strain NEAU-gxj3Tcould be differentiated clearly from its closest phylogenetic relatives. Therefore, the strain is concluded to represent a novel species of the genus Plantactinospora, for which the name Plantactinospora soyae sp. nov. is proposed. The type strain is NEAU-gxj3T (=CGMCC 4.7221T=DSM 46832T).

  13. Sphaerisporangium dianthi sp. nov., an endophytic actinomycete isolated from a root of Dianthus chinensis L.

    PubMed

    Xing, Jia; Liu, Chongxi; Zhang, Yuejing; He, Hairong; Zhou, Ying; Li, Lianjie; Zhao, Junwei; Liu, Shuanghe; Wang, Xiangjing; Xiang, Wensheng

    2015-01-01

    A novel actinomycete, designated strain NEAU-CY18(T), was isolated from the root of a Chinese medicinal plant Dianthus chinensis L and subjected to a polyphasic taxonomic study. The novel strain was found to develop spherical sporangia with non-motile spores on aerial mycelium. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The whole-cell sugars were identified as madurose, mannose, ribose, galactose and glucose. The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and an unidentified phospholipid. The predominant menaquinones were identified as MK-9(H4), MK-9(H2) and MK-9(H6). The major fatty acids were identified as C17:0 10-methyl, iso-C16:0 and C16:0. EzTaxon-e analysis of the 16S rRNA gene sequence indicated that the strain belongs to the genus Sphaerisporangium and was most closely related to Sphaerisporangium cinnabarinum JCM 3291(T) (98.9 %) and Sphaerisporangium melleum JCM 13064(T) (98.3 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-CY18(T) forms a monophyletic clade with S. cinnabarinum JCM 3291(T), an association that was supported by a bootstrap value of 97 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. Comparisons of some phenotypic properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. cinnabarinum JCM 3291(T) and S. melleum JCM 13064(T). Therefore, it is concluded that strain NEAU-CY18(T) represents a novel Sphaerisporangium species, for which the name Sphaerisporangium dianthi sp. nov. is proposed. The type strain is NEAU-CY18(T) ( = CGMCC 4.7132(T) = DSM 46736(T)).

  14. Micromonospora zeae sp. nov., a novel endophytic actinomycete isolated from corn root (Zea mays L.).

    PubMed

    Shen, Yue; Zhang, Yuejing; Liu, Chongxi; Wang, Xiangjing; Zhao, Junwei; Jia, Feiyu; Yang, Lingyu; Yang, Deguang; Xiang, Wensheng

    2014-11-01

    A novel actinomycete, designated strain NEAU-gq9(T), was isolated from corn root (Zea mays L.) and characterized using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of the genus Micromonospora. On the basis of 16S rRNA gene sequence similarity studies, strain NEAU-gq9(T) was most closely related to Micromonospora zamorensis CR38(T) (99.3%), Micromonospora jinlongensis NEAU-GRX11(T) (99.2%), Micromonospora saelicesensis Lupac 09(T) (99.2%), Micromonospora chokoriensis 2-19(6)(T) (98.9%), Micromonospora coxensis 2-30-b(28)(T) (98.6%) and Micromonospora lupini Lupac 14N(T) (98.5%). Phylogenetic analysis based on the 16S rRNA gene and gyrB gene demonstrated that strain NEAU-gq9(T) is a member of the genus Micromonospora and supported the closest phylogenetic relationship to M. zamorensis CR38(T), M. jinlongensis NEAU-GRX11(T), M. saelicesensis Lupac 09(T), M. chokoriensis 2-19(6)(T) and M. lupini Lupac 14N(T). A combination of DNA-DNA hybridization, morphological and physiological characteristics indicated that the novel strain could be readily distinguished from the closest phylogenetic relatives. Therefore, it is proposed that strain NEAU-gq9(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora zeae sp. nov. is proposed. The type strain is NEAU-gq9(T) (=CGMCC 4.7092(T)=DSM 45882(T)).

  15. Biosynthetic potential of phylogenetically unique endophytic actinomycetes from tropical plants.

    PubMed

    Janso, Jeffrey E; Carter, Guy T

    2010-07-01

    The culturable diversity of endophytic actinomycetes associated with tropical, native plants is essentially unexplored. In this study, 123 endophytic actinomycetes were isolated from tropical plants collected from several locations in Papua New Guinea and Mborokua Island, Solomon Islands. Isolates were found to be prevalent in roots but uncommon in leaves. Initially, isolates were dereplicated to the strain level by ribotyping. Subsequent characterization of 105 unique strains by 16S rRNA gene sequence analysis revealed that 17 different genera were represented, and rare genera, such as Sphaerisporangium and Planotetraspora, which have never been previously reported to be endophytic, were quite prevalent. Phylogenetic analyses grouped many of the strains into clades distinct from known genera within Thermomonosporaceae and Micromonosporaceae, indicating that they may be unique genera. Bioactivity testing and liquid chromatography-mass spectrometry (LC-MS) profiling of crude fermentation extracts were performed on 91 strains. About 60% of the extracts exhibited bioactivity or displayed LC-MS profiles with spectra indicative of secondary metabolites. The biosynthetic potential of 29 nonproductive strains was further investigated by the detection of putative polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) genes. Despite their lack of detectable secondary metabolite production in fermentation, most were positive for type I (66%) and type II (79%) PKS genes, and all were positive for NRPS genes. These results suggest that tropical plants from New Guinea and the adjacent archipelago are hosts to unique endophytic actinomycetes that possess significant biosynthetic potential.

  16. Nonomuraea syzygii sp. nov., an endophytic actinomycete isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels).

    PubMed

    Rachniyom, Hathairat; Matsumoto, Atsuko; Indananda, Chantra; Duangmal, Kannika; Takahashi, Yoko; Thamchaipenet, Arinthip

    2015-04-01

    A novel endophytic actinomycete, designated strain GKU 164(T), was isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels), collected at Khao Khitchakut National Park, Chantaburi province, Thailand. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain formed a distinct clade within the genus Nonomuraea , and was most closely related to Nonomuraea monospora PT708(T) (98.77% 16S rRNA gene sequence similarity) and Nonomuraea thailandensis KC-061(T) (98.73%). Strain GKU 164(T) formed a branched substrate and aerial hyphae that generated single spores with rough surfaces. The cell wall contained meso-diaminopimelic acid. The whole-cell sugars were madurose, galactose, mannose, ribose, rhamnose and glucose. The N-acyl type of muramic acid was acetyl. The predominant menaquinone was MK-9(H4) with minor amounts of MK-9(H6), MK-9(H2) and MK-9(H0). The phospholipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositolmannosides, phosphatidylmonomethylethanolamine, hydroxy-phosphatidylmonomethylethanolamine, an unidentified aminophosphoglycolipid and four unknown phospholipids. The major fatty acids were iso-C(16 : 0) and 10-methyl C(17 : 0). The genomic DNA G+C content was 70.4 mol%. Significant differences in the morphological, chemotaxonomical, and biochemical data together with DNA-DNA relatedness values between strain GKU 164(T) and type strains of closely related species, clearly demonstrated that strain GKU 164(T) represents a novel species of the genus Nonomuraea , for which the name Nonomuraea syzygii sp. nov. is proposed. The type strain is GKU 164(T) ( = BCC 70457(T) = NBRC 110400(T)). © 2015 IUMS.

  17. Actinomadura syzygii sp. nov., an endophytic actinomycete isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels).

    PubMed

    Rachniyom, Hathairat; Matsumoto, Atsuko; Indananda, Chantra; Duangmal, Kannika; Takahashi, Yoko; Thamchaipenet, Arinthip

    2015-06-01

    The taxonomic position of an endophytic actinomycete, strain GKU 157T, isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels) collected at Khao Khitchakut National Park, Chantaburi province, Thailand, was determined using a polyphasic taxonomic approach. 16S rRNA gene sequence analysis revealed that strain GKU 157T belongs to the genus Actinomadura and formed a distinct phyletic line with Actinomadura chibensis NBRC 106107T (98.6 % similarity). Strain GKU 157T formed an extensively branched, non-fragmenting substrate mycelium and aerial hyphae that differentiated into hooked to short spiral chains of about 20 non-motile spores with a warty surface. The cell wall contained meso-diaminopimelic acid and the whole-cell sugars were galactose, glucose, madurose, mannose and ribose. The N-acyl type of muramic acid was acetyl. Mycolic acids were absent. The phospholipids included phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylinositol (PI), phosphatidylinositolmannoside (PIM) and two unknown phospholipids (PLs). The major menaquinone was MK-9(H6) and the predominant fatty acids were C16:0, iso-C16:0, C18:1ω9c, C18:0 and 10-methyl C18:0 (tuberculostearic acid). The genomic DNA G+C content was 73.1 mol%. A combination of DNA-DNA hybridization results and significant differences from related species in cultural, physiological and chemical characteristics indicated that strain GKU 157T represents a novel species of the genus Actinomadura, for which the name Actinomadura syzygii sp. nov. is proposed. The type strain is GKU 157T ( = BCC 70456T = NBRC 110399T).

  18. Microbispora sp. LGMB259 endophytic actinomycete isolated from Vochysia divergens (Pantanal, Brazil) producing β-carbolines and indoles with biological activity.

    PubMed

    Savi, Daiani C; Shaaban, Khaled A; Vargas, Nathalia; Ponomareva, Larissa V; Possiede, Yvelise M; Thorson, Jon S; Glienke, Chirlei; Rohr, Jürgen

    2015-03-01

    Endophytic actinomycetes encompass bacterial groups that are well known for the production of a diverse range of secondary metabolites. Vochysia divergens is a medicinal plant, common in the "Pantanal" region (Brazil) and was focus of many investigations, but never regarding its community of endophytic symbionts. During a screening program, an endophytic strain isolated from the V. divergens, was investigated for its potential to show biological activity. The strain was characterized as Microbispora sp. LGMB259 by spore morphology and molecular analyze using nucleotide sequence of the 16S rRNA gene. Strain LGMB259 was cultivated in R5A medium producing metabolites with significant antibacterial activity. The strain produced 4 chemically related β-carbolines, and 3 Indoles. Compound 1-vinyl-β-carboline-3-carboxylic acid displayed potent activity against the Gram-positive bacterial strains Micrococcus luteus NRRL B-2618 and Kocuria rosea B-1106, and was highly active against two human cancer cell lines, namely the prostate cancer cell line PC3 and the non-small-cell lung carcinoma cell line A549, with IC50 values of 9.45 and 24.67 µM, respectively. 1-Vinyl-β-carboline-3-carboxylic acid also showed moderate activity against the yeast Saccharomyces cerevisiae ATCC204508, as well as the phytopathogenic fungi Phyllosticta citricarpa LGMB06 and Colletotrichum gloeosporioides FDC83.

  19. Microbispora sp. LGMB259 Endophytic Actinomycete Isolated from Vochysia divergens (Pantanal, Brazil) Producing β-Carbolines and Indoles with Biological Activity

    PubMed Central

    Savi, Daiani C.; Shaaban, Khaled A.; Vargas, Nathalia; Ponomareva, Larissa V.; Possiede, Yvelise M.; Thorson, Jon S.; Glienke, Chirlei; Rohr, Jürgen

    2014-01-01

    Endophytic actinomycetes encompass bacterial groups that are well known for the production of a diverse range of secondary metabolites. Vochysia divergens is a medicinal plant, common in the “Pantanal” region (Brazil) and was focus of many investigations, but never regarding its community of endophytic symbionts. During a screening program, an endophytic strain isolated from the V. divergens, was investigated for its potential to show biological activity. The strain was characterized as Microbispora sp. LGMB259 by spore morphology and molecular analyze using nucleotide sequence of the 16S rRNA gene. Strain LGMB259 was cultivated in R5A medium producing metabolites with significant antibacterial activity. The strain produced 4 chemically related β-carbolines, and 3 Indoles. Compound 1-Vinyl-β-carboline-3-carboxylic acid displayed potent activity against the Gram-positive bacterial strains Micrococcus luteus NRRL B-2618 and Kocuria rosea B-1106, and was highly active against two human cancer cell lines, namely the prostate cancer cell line PC3 and the non-small-cell lung carcinoma cell line A549, with IC50 values of 9.45 and 24.67 µM, respectively. 1-Vinyl-β-carboline-3-carboxylic acid also showed moderate activity against the yeast Saccharomyces cerevisiae ATCC204508, as well as the phytopathogenic fungiPhyllosticta citricarpa LGMB06 and Colletotrichum gloeosporioides FDC83. PMID:25385358

  20. Nocardioides panzhihuaensis sp. nov., a novel endophytic actinomycete isolated from medicinal plant Jatropha curcas L.

    PubMed

    Qin, Sheng; Yuan, Bo; Zhang, Yue-Ji; Bian, Guang-Kai; Tamura, Tomohiko; Sun, Bing-Zhi; Li, Wen-Jun; Jiang, Ji-Hong

    2012-08-01

    A novel Gram-positive, aerobic, rod-shaped and mycelia-producing bacterial strain, designated KLBMP 1050(T), was isolated from the stem of the oil-seed plant Jatropha curcas L. collected from Sichuan Province, south-west China. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the isolate KLBMP 1050(T) belonged to the genus Nocardioides, with the highest sequence similarity to Nocardioides albus KCTC 9186(T) (99.38 %) and Nocardioides luteus KCTC 9575(T) (99.03 %). However, the DNA-DNA relatedness of isolate KLBMP 1050(T) to these two type strains were 37.5 ± 3.5 and 33 ± 2.3 %, respectively. Strain KLBMP 1050(T) grew at the pH range 6-11, temperature range 10-32 °C and with 0-12 % NaCl. The physiological properties of strain KLBMP 1050(T) differ from those of N. albus KCTC 9186(T) and N. luteus KCTC 9575(T). The cell-wall peptidoglycan contained LL: -diaminopimelic acid and MK-8(H(4)) was the major respiratory quinone. The predominant cellular fatty acid of strain KLBMP 1050(T) was iso-C(16:0) (23.3 %). The total DNA G+C content was 70.1 mol%. On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain KLBMP 1050(T) represents a novel species of the genus Nocardioides, for which the name Nocardioides panzhihuaensis sp. nov. is proposed. The type strain is KLBMP 1050(T) (= KCTC 19888(T) = NBRC 108680(T)).

  1. Pseudonocardia nantongensis sp. nov., a novel endophytic actinomycete isolated from the coastal halophyte Tamarix chinensis Lour.

    PubMed

    Xing, Ke; Qin, Sheng; Bian, Guang-Kai; Zhang, Yue-Ji; Zhang, Wen-Di; Dai, Chuan-Chao; Liu, Chang-Hong; Li, Wen-Jun; Jiang, Ji-Hong

    2012-11-01

    A novel isolate, designated strain KLBMP 1282(T) was isolated from the surface-sterilized leaves of a coastal halophyte Tamarix chinensis Lour., collected from Nantong, Jiangsu Province, east of China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that this strain belongs to the genus Pseudonocardia, being most closely related to Pseudonocardia kongjuensis LM 157(T) (98.33 %), Pseudonocardia autotrophica IMSNU 20050(T) (97.77 %), Pseudonocardia endophytica YIM 56035(T) (97.63 %), Pseudonocardia ammonioxydans H9 (T) (97.62 %) and Pseudonocardia compacta IMSNU 20111(T) (97.56 %); similarity to other type strains of the genus Pseudonocardia was <97.5 %. Chemotaxonomic data confirmed the affiliation of strain KLBMP 1282(T) to the genus Pseudonocardia. Strain KLBMP 1282(T) contained MK-8(H(4)) as the predominant ubiquinone and iso-C(16:0) as the major fatty acid. The polar lipids detected in strain KLBMP 1282(T) were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, one unknown phospholipid and four unknown glycolipids. The DNA G + C content of strain KLBMP 1282(T) was 73.1 mol %. The results of DNA-DNA hybridizations and the phylogenetic analysis, together with the phenotypic and biochemical tests, allowed the differentiation of strain KLBMP 1282(T) from strains of other recognized Pseudonocardia species. Therefore, strain KLBMP 1282(T) represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia nantongensis sp. nov. is proposed. The type strain is KLBMP 1282(T) (=KCTC 29053(T) = NBRC 108677(T)).

  2. Biocontrol of Rhizoctonia solani damping-off and promotion of tomato plant growth by endophytic actinomycetes isolated from native plants of Algerian Sahara.

    PubMed

    Goudjal, Yacine; Toumatia, Omrane; Yekkour, Amine; Sabaou, Nasserdine; Mathieu, Florence; Zitouni, Abdelghani

    2014-01-20

    Thirty-four endophytic actinomycetes were isolated from the roots of native plants of the Algerian Sahara. Morphological and chemical studies showed that twenty-nine isolates belonged to the Streptomyces genus and five were non-Streptomyces. All isolates were screened for their in vitro antifungal activity against Rhizoctonia solani. The six that had the greatest pathogen inhibitory capacities were subsequently tested for their in vivo biocontrol potential on R. solani damping-off in sterilized and non-sterilized soils, and for their plant-growth promoting activities on tomato seedlings. In both soils, coating tomato seeds with antagonistic isolates significantly reduced (P<0.05) the severity of damping-off of tomato seedlings. Among the isolates tested, the strains CA-2 and AA-2 exhibited the same disease incidence reduction as thioperoxydicarbonic diamide, tetramethylthiram (TMTD) and no significant differences (P<0.05) were observed. Furthermore, they resulted in a significant increase in the seedling fresh weight, the seedling length and the root length of the seed-treated seedlings compared to the control. The taxonomic position based on 16S rDNA sequence analysis and phylogenetic studies indicated that the strains CA-2 and AA-2 were related to Streptomyces mutabilis NBRC 12800(T) (100% of similarity) and Streptomyces cyaneofuscatus JCM 4364(T) (100% of similarity), respectively.

  3. Continuing hunt for endophytic actinomycetes as a source of novel biologically active metabolites.

    PubMed

    Masand, Meeta; Jose, Polpass Arul; Menghani, Ekta; Jebakumar, Solomon Robinson David

    2015-12-01

    Drug-resistant pathogens and persistent agrochemicals mount the detrimental threats against human health and welfare. Exploitation of beneficial microorganisms and their metabolic inventions is most promising way to tackle these two problems. Since the successive discoveries of penicillin and streptomycin in 1940s, numerous biologically active metabolites have been discovered from different microorganisms, especially actinomycetes. In recent years, actinomycetes that inhabit unexplored environments have received significant attention due to their broad diversity and distinctive metabolic potential with medical, agricultural and industrial importance. In this scenario, endophytic actinomycetes that inhabit living tissues of plants are emerging as a potential source of novel bioactive compounds for the discovery of drug leads. Also, endophytic actinomycetes are considered as bio-inoculants to improve crop performance through organic farming practices. Further efforts on exploring the endophytic actinomycetes associated with the plants warrant the likelihood of discovering new taxa and their metabolites with novel chemical structures and biotechnological importance. This mini-review highlights the recent achievements in isolation of endophytic actinomycetes and an assortment of bioactive compounds.

  4. [Bioactivity of endophytic actinomycetes from medicinal plants and secondary metabolites from strain D62].

    PubMed

    Liu, Ning; Zhang, Hui; Zheng, Wen; Huang, Ying; Wang, Hai-Bin

    2007-10-01

    It is believed that genetic recombination of the endophytes with the hosts that occurred in evolutionary time could result in some endophytes producing certain phytochemical originally characteristic of the host. Based on this widely accepted hypothesis, there have been increasing research efforts focused on screening for novel natural products from endophytes. In this study, antimicrobial and antitumor activities of 165 actinomycetes isolated from medicinal plants collected from Xishuangbanna were tested by agar diffusion method and WST-8 assay respectively. The results showed that over 42% of the isolates exhibited antagonism against pathogenic strains, and 54.5% displayed excellent inhibition against mouse melanoma cell line B16 or/and human alveolar epithelial cell line A549. These results are superior to those of soil actinomycetes, indicating tremendous potential of endophytic of actinomycetes for exploration. Six compounds that had both antimicrobial and antitumor activities were separated and purified from isolate Streptomyces sp. D62 by resin adsorption, silica-gel column and sephadex chromatography, etc. On the basis of spectral analyses, they were identified as antimycin A4a (1), antimycin A7a (2), antimycin A2a (3), antimycin A1a (4), 10-hydroxy-10-methyl-dodec-2-en-1,4-olide (5) and 6-(2-(4-aminophenyl)-2-oxoethyl)-3,5-dimethyl-tetrahydropyran-2-one(6), with the last one defined as a novel compound. Based on all these results, it is convinced that endophytic actinomycetes are a promising resource for bioactive natural product discovery.

  5. Larvicidal potential of Asteraceae family endophytic actinomycetes against Culex quinquefasciatus mosquito larvae.

    PubMed

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida

    2014-01-01

    Pakistan is blessed with plants of Asteraceae family with known medicinal background used for centuries by Hakims (traditional physicians). Keeping in mind the background of their anti-larval potential, a total of 21 endophytic actinomycetes were isolated from four Asteraceae plants and screened against the first and fourth instar stages of Culex quinquefasciatus Say mosquito larvae. Of the 21 isolates, 6 of them gave strong larvicidal activity (80-100% mortality) in the screening results and 4 isolates gave a potent larvicidal activity (100% mortality) at the fourth instar stage. These isolates belonged to different species within the actinomycetes group, namely Streptomyces albovinaceus and Streptomyces badius. This communication reports the larvicidal potential of endophytic actinomycetes residing within the native Asteraceae plants in Pakistan. The study suggests further exploration through large-scale productions leading to the identification of the larvicidal compounds.

  6. Antimicrobial biosynthetic potential and genetic diversity of endophytic actinomycetes associated with medicinal plants.

    PubMed

    Gohain, Anwesha; Gogoi, Animesh; Debnath, Rajal; Yadav, Archana; Singh, Bhim P; Gupta, Vijai K; Sharma, Rajeev; Saikia, Ratul

    2015-10-01

    Endophytic actinomycetes are one of the primary groups that share symbiotic relationships with medicinal plants and are key reservoir of biologically active compounds. In this study, six selective medicinal plants were targeted for the first time for endophytic actinomycetes isolation from Gibbon Wild Life Sanctuary, Assam, India, during winter and summer and 76 isolates were obtained. The isolates were found to be prevalent in roots followed by stem and leaves. 16S rRNA gene sequence analysis revealed 16 genera, including rare genera, Verrucosispora, Isoptericola and Kytococcus, which have never been previously reported as endophytic. The genus Streptomyces (66%) was dominant in both seasons. Shannon's diversity index showed that Azadirachta indica (1.49), Rauwolfia serpentina (1.43) and Emblica officinalis (1.24) were relatively good habitat for endophytic actinomycetes. Antimicrobial strains showed prevalence of polyketide synthase (PKS) type-II (85%) followed by PKS type-I (14%) encoded in the genomes. Expression studies showed 12-fold upregulation of PKSII gene in seventh day of incubation for Streptomyces antibioticus (EAAG90). Our results emphasize that the actinomycetes assemblages within plant tissue exhibited biosynthetic systems encoding for important biologically active compounds.

  7. Micromonospora taraxaci sp. nov., a novel endophytic actinomycete isolated from dandelion root (Taraxacum mongolicum Hand.-Mazz.).

    PubMed

    Zhao, Junwei; Guo, Lifeng; He, Hairong; Liu, Chongxi; Zhang, Yuejing; Li, Chuang; Wang, Xiangjing; Xiang, Wensheng

    2014-10-01

    A novel actinomycete, designated strain NEAU-P5(T), was isolated from dandelion root (Taraxacum mongolicum Hand.-Mazz.). Strain NEAU-P5(T) showed closest 16S rRNA gene sequence similarity to Micromonospora chokoriensis 2-19/6(T) (99.5%), and phylogenetically clustered with Micromonospora violae NEAU-zh8(T) (99.3%), M. saelicesensis Lupac 09(T) (99.0%), M. lupini Lupac 14N(T) (98.8%), M. zeae NEAU-gq9(T) (98.4%), M. jinlongensis NEAU-GRX11(T) (98.3%) and M. zamorensis CR38(T) (97.9%). Phylogenetic analysis based on the gyrB gene sequence also indicated that the isolate clustered with the above type strains except M. violae NEAU-zh8(T). The cell-wall peptidoglycan consisted of meso-diaminopimelic acid and glycine. The major menaquinones were MK-9(H8), MK-9(H6) and MK-10(H2). The phospholipid profile contained diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The major fatty acids were C(16:0), iso-C(15:0) and C(17:0). Furthermore, some physiological and biochemical properties and low DNA-DNA relatedness values enabled the strain to be differentiated from members of closely related species. Therefore, it is proposed that strain NEAU-P5(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora taraxaci sp. nov. is proposed. The type strain is NEAU-P5(T) (=CGMCC 4.7098(T) = DSM 45885(T)).

  8. Generic and functional diversity in endophytic actinomycetes from wild Compositae plant species at South Sinai - Egypt.

    PubMed

    El-Shatoury, Sahar A; El-Kraly, Omnia A; Trujillo, Martha E; El-Kazzaz, Waleed M; El-Din, El-Sayeda Gamal; Dewedar, Ahmed

    2013-09-01

    The diversity of culturable endophytic actinomycetes associated with wild Compositae plants is scantily explored. In this study, one hundred and thirty one endophytic actinobacteria were isolated from ten Compositae plant species collected from South Sinai in Egypt. Microscopic and chemotaxonomic investigation of the isolates indicated fourteen genera. Rare genera, such as Microtetraspora, and Intrasporangium, which have never been previously reported to be endophytic, were identified. Each plant species accommodated between three to eight genera of actinobacteria and unidentified strains were recovered from seven plant species. The generic diversity analysis of endophytic assemblages grouped the plant species into three main clusters, representing high, moderate and low endophytic diversity. The endophytes showed high functional diversity, based on forty four catabolic and plant growth promotion traits; providing some evidence that such traits could represent key criteria for successful residence of endophytes in the endosphere. Stress-tolerance traits were more predictive measure of functional diversity differences between the endophyte assemblages (Shannon's index, p = 0.01). The results indicate a potential prominent role of endophytes for their hosts and emphasize the potency of plant endosphere as a habitat for actinobacteria with promising future applications.

  9. Biosynthetic Potential of Phylogenetically Unique Endophytic Actinomycetes from Tropical Plants▿ †

    PubMed Central

    Janso, Jeffrey E.; Carter, Guy T.

    2010-01-01

    The culturable diversity of endophytic actinomycetes associated with tropical, native plants is essentially unexplored. In this study, 123 endophytic actinomycetes were isolated from tropical plants collected from several locations in Papua New Guinea and Mborokua Island, Solomon Islands. Isolates were found to be prevalent in roots but uncommon in leaves. Initially, isolates were dereplicated to the strain level by ribotyping. Subsequent characterization of 105 unique strains by 16S rRNA gene sequence analysis revealed that 17 different genera were represented, and rare genera, such as Sphaerisporangium and Planotetraspora, which have never been previously reported to be endophytic, were quite prevalent. Phylogenetic analyses grouped many of the strains into clades distinct from known genera within Thermomonosporaceae and Micromonosporaceae, indicating that they may be unique genera. Bioactivity testing and liquid chromatography-mass spectrometry (LC-MS) profiling of crude fermentation extracts were performed on 91 strains. About 60% of the extracts exhibited bioactivity or displayed LC-MS profiles with spectra indicative of secondary metabolites. The biosynthetic potential of 29 nonproductive strains was further investigated by the detection of putative polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) genes. Despite their lack of detectable secondary metabolite production in fermentation, most were positive for type I (66%) and type II (79%) PKS genes, and all were positive for NRPS genes. These results suggest that tropical plants from New Guinea and the adjacent archipelago are hosts to unique endophytic actinomycetes that possess significant biosynthetic potential. PMID:20472734

  10. [Selective Isolation of Rare Actinomycetes from Soil].

    PubMed

    Sineva, O N; Terekhova, L P

    2015-01-01

    Many diverse methods for selective isolation of actinomycetes are used in discovery of organisms producing biologically active substances, as well as in ecological studies. Methods for isolation of rare actinomycetes from soil are reviewed.

  11. Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv. glycine.

    PubMed

    Mingma, Ratchanee; Pathom-aree, Wasu; Trakulnaleamsai, Savitr; Thamchaipenet, Arinthip; Duangmal, Kannika

    2014-01-01

    In this study, actinomycetes from roots and rhizospheric soils of leguminous plants were isolated using starch casein agar supplemented with antifungal and antibacterial antibiotics. Three hundred and seventeen actinomycetes were isolated with 77 isolates obtained from plant roots and 240 isolates from rhizospheric soils. Analysis of whole-organism hydrolysates showed that 289 strains were rich in the LL-isomer of diaminopimelic acid, a result consistent with their assignment to the streptomycetes. The remaining 28 strains were assigned to non-streptomycetes based on the presence of meso-isomer of diaminopimelic acid in cell wall. Sixty-four isolates (20.2%) showed antagonistic activity against soybean pathogen Xanthomonas campestris pv. glycine by agar overlay method. Isolate RM 365 showed the highest activity with an inhibition ratio of 3.79, with no inhibitory activity on the growth of Rhizobium japonicum TISTR 079, Rhizobium sp. TISTR 061 and Rhizobium sp. TISTR 063. The 16S rRNA gene sequence analysis revealed that isolate RM 365 shared 99.28% similarity to Streptomyces caeruleatus GIMN4(T) (GQ329712). In addition, isolates which contained meso-DAP were also identified by 16S rRNA gene sequence analysis. The results showed that they were members of the genus Amycolatopsis, Isoptericola, Micromonospora, Microbispora, Nocardia, Nonomuraea, Promicromonospora and Pseudonocardia.

  12. Novel Actinomycete Isolated from Bulking Industrial Sludge

    PubMed Central

    White, Johanna M.; Labeda, David P.; Lechevalier, Mary P.; Owens, James R.; Jones, Daniel D.; Gauthier, Joseph J.

    1986-01-01

    A novel actinomycete was the predominant filamentous microorganism in bulking activated sludge in a bench-scale reactor treating coke plant wastewater. The bacterium was isolated and identified as an actinomycete that is biochemically and morphologically similar to Amycolatopsis orientalis; however, a lack of DNA homology excludes true relatedness. At present, the isolate (NRRL B 16216) cannot be assigned to the recognized taxa of actinomycetes. Images PMID:16347238

  13. Identification and screening of rare actinomycetes isolated from Neesia altissima Bl.

    NASA Astrophysics Data System (ADS)

    Pratiwi, R. H.; Hidayat, I.; Hanafi, M.; Mangunwardoyo, W.

    2017-07-01

    Actinomycetes is the main source of antibiotics and endophytic actinomycetes from medicinal plants has considerable potential as like the host. The aim of this research is to identify rare actinomycetes isolated from Neesia altissima and to screen their antagonistic activity against diarrhea-causing bacteria in order to find new potential secondary metabolites. Samples of N. altissima were collected from mount Halimun-Salak National Park. Endophytic actinomycetes were isolated from roots of N. altissima by surface sterilization method. Screening of antagonistic activity was conducted against five diarrhea-causing bacteria such as Bacillus cereus ATCC 10876, Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 25241, Shigella flexneri ATCC 12022, and Staphylococcus aureus ATCC 25923 by using diffusion disc methods. The endophytic actinomycete showed in vitro antibacterial activity against four diarrhea-causing bacteria, except the B. cereus ATCC 10876. The phylogenetic tree generated from 16S rRNA sequence showed that sequence of endophytic actinomycetes isolates nested in the clade belonging to the genus Nonomuraea. Sequence of UICC B-94 formed a monophyletic clade with N. jabiensis strain A4036 and N. rubra strain AC 615. Therefore, it is named as Nonomuraea sp. strain UICC B-94.

  14. Isolation of cellulolytic actinomycetes from marine sediments

    SciTech Connect

    Veiga, M.; Esparis, A.; Fabregas, J.

    1983-07-01

    The cellulolytic activity of 36 actinomycetes strains isolated from marine sediments was investigated by the cellulose-azure method. Approximately 50% of the isolates exhibited various degrees of cellulolytic activity. 13 references.

  15. Streptomyces halophytocola sp. nov., an endophytic actinomycete isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour.

    PubMed

    Qin, Sheng; Bian, Guang-Kai; Tamura, Tomohiko; Zhang, Yue-Ji; Zhang, Wen-Di; Cao, Cheng-Liang; Jiang, Ji-Hong

    2013-08-01

    A novel actinomycete, designated KLBMP 1284(T), was isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour. collected from the city of Nantong, Jiangsu Province, east China. The strain was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Analysis of the 16S rRNA gene sequence of strain KLBMP 1284(T) revealed that the strain formed a distinct clade within the phylogenetic tree based on 16S rRNA gene sequences and the highest sequence similarity (99.43 %) was to Streptomyces sulphureus NRRL B-1627(T). 16S rRNA gene sequence similarity to other species of the genus Streptomyces was lower than 97 %. Based on DNA-DNA hybridization values and comparison of morphological and phenotypic data, KLBMP 1284(T) could be distinguished from the closest phylogenetically related species, Streptomyces sulphureus NRRL B-1627(T). Thus, based on these data, it is evident that strain KLBMP 1284(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces halophytocola sp. nov. is proposed. The type strain is KLBMP 1284(T) (= KCTC 19890(T) = NBRC 108770(T)).

  16. Evaluation of antagonistic and plant growth promoting activities of chitinolytic endophytic actinomycetes associated with medicinal plants against Sclerotium rolfsii in chickpea.

    PubMed

    Singh, S P; Gaur, R

    2016-08-01

    To evaluate the potential of chitinolytic endophytic Actinomycetes isolated from medicinal plants in order to diminish the collar rot infestation induced by Sclerotium rolfsii in chickpea. Sixty-eight chitinolytic endophytic Actinomycetes were recovered from various medicinal plants and evaluated for their chitinase activity. Among these isolates, 12 were screened for their plant growth promoting abilities and antagonistic potential against Sc. rolfsii. Further, these isolates were validated in vivo for their ability to protect chickpea against Sc. rolfsii infestation under greenhouse conditions. The isolates significantly (P < 0·05) increased the biomass (1·2-2·0 fold) and reduced plant mortality (42-75%) of chickpea. On the basis of 16S rDNA profiling, the selected antagonistic strains were identified as Streptomyces diastaticus, Streptomyces fradiae, Streptomyces olivochromogenes, Streptomyces collinus, Streptomyces ossamyceticus and Streptomyces griseus. This study is the first report of the isolation of endophytic Actinomycetes from various medicinal plants having antagonistic and plant growth promoting abilities. The isolated species showed potential for controlling collar rot disease on chickpea and could be useful in integrated control against diverse soil borne plant pathogens. Our investigation suggests that endophytic Actinomycetes associated with medicinal plants can be used as bioinoculants for developing safe, efficacious and environment-friendly biocontrol strategies in the near future. © 2016 The Society for Applied Microbiology.

  17. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants

    PubMed Central

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Gupta, Vijai Kumar; Yadav, Mukesh Kumar; Saikia, Ratul; Singh, Bhim Pratap

    2015-01-01

    Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10–32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these

  18. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

    PubMed

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Gupta, Vijai Kumar; Yadav, Mukesh Kumar; Saikia, Ratul; Singh, Bhim Pratap

    2015-01-01

    Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these

  19. Biogenic gold nanotriangles from Saccharomonospora sp., an endophytic actinomycetes of Azadirachta indica A. Juss.

    NASA Astrophysics Data System (ADS)

    Verma, Vijay C.; Anand, Swechha; Ulrichs, Christian; Singh, Santosh K.

    2013-04-01

    Microbial biofabrication is emerging as eco-friendly, simpler, and reproducible alternative to chemical synthesis of metals and semiconductor nanoparticles, allowing generation of rare geometrical forms such as nanotriangles and nanoprisms. Highly confined nanostructures like triangles/prisms are interesting class of nanoparticles due to their unique optical properties exploitable in biomedical diagnostics and biosensors. Here, we report for the first time a single-step biological protocol for the synthesis of gold nanotriangles using extract of endophytic actinomycetes Saccharomonospora sp., isolated from surface sterilized root tissues of Azadirachta indica A. Juss., when incubated with an aqueous solution of chloroaurate ions (AuCl- 4/1 mM). Thin, flat occasionally prismatic gold nanotriangles were produced when aqueous chloroaurate ions reacted with the cell-free extract as well as with the biomass of endophytic Saccharomonospora. It was evidenced from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis that proteins of 42 and 50 kD were involved in biosynthesis as well as in stabilization of the nanoparticles. The particle growth process was monitored by UV-vis spectroscopy, and the morphological characterization was carried out by transmission electron microscopy and atomic force microscopy together with X-ray powder diffractions. Although the exact mechanism for this shape-oriented synthesis is not clear so far, the possibility of achieving nanoparticle shape control in a microbial system is exciting.

  20. Culturable rare Actinomycetes: diversity, isolation and marine natural product discovery.

    PubMed

    Subramani, Ramesh; Aalbersberg, William

    2013-11-01

    Rare Actinomycetes from underexplored marine environments are targeted in drug discovery studies due to the Actinomycetes' potentially huge resource of structurally diverse natural products with unusual biological activity. Of all marine bacteria, 10 % are Actinomycetes, which have proven an outstanding and fascinating resource for new and potent bioactive molecules. Past and present efforts in the isolation of rare Actinomycetes from underexplored diverse natural habitats have resulted in the isolation of about 220 rare Actinomycete genera of which more than 50 taxa have been reported to be the producers of 2,500 bioactive compounds. That amount represents greater than 25 % of the total Actinomycetes metabolites, demonstrating that selective isolation methods are being developed and extensively applied. Due to the high rediscovery rate of known compounds from Actinomycetes, a renewed interest in the development of new antimicrobial agents from rare and novel Actinomycetes is urgently required to combat the increasing number of multidrug-resistant human pathogens. To facilitate that discovery, this review updates all selective isolation media including pretreatment and enrichment methods for the isolation of marine rare Actinomycetes. In addition, this review demonstrates that discovering new compounds with novel scaffolds can be increased by intensive efforts in isolating and screening rare marine genera of Actinomycetes. Between 2007 and mid-2013, 80 new rare Actinomycete species were reported from marine habitats. They belong to 23 rare families, of which three are novel, and 20 novel genera. Of them, the family Micromonosporaceae is dominant as a producer of promising chemical diversity.

  1. Pseudonocardia antimicrobica sp. nov., a novel endophytic actinomycete associated with Artemisia annua L. (sweet wormwood).

    PubMed

    Zhao, Guo-Zhen; Li, Jie; Qin, Yu-Li; Miao, Cui-Ping; Wei, Da-Qiao; Zhang, Si; Xu, Li-Hua; Li, Wen-Jun

    2012-09-01

    A Gram-reaction-positive, non-motile, endophytic actinomycete, designated strain YIM 63235(T), was isolated from the surface-sterilized stems of Artemisia annua L., and characterized to determine its taxonomic position. The strain YIM 63235(T) formed well-differentiated aerial and substrate mycelia on media tested. The phylogenetic tree based on 16S rRNA gene sequences showed that the new isolate formed a distinct lineage within the genus Pseudonocardia, and the strain YIM 63235(T) was closely related to Pseudonocardia parietis 04-St-002(T) (99.1%). However, DNA-DNA relatedness demonstrated that strain YIM 63235(T) was distinct from the closest phylogenetic neighbor. The chemotaxonomic properties of strain YIM 63235(T) were consistent with those of the genus Pseudonocardia: the diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid and MK-8(H(4)) was the predominant menaquinone. The major fatty acids were iso-C(16:0) and iso-C(16:1) H. The DNA G+C content of strain YIM 63235(T) was 71.0 mol%. On the basis of the phenotypic and phylogenetic distinctiveness, the novel isolate was identified as representing a novel species of the genus Pseudonocardia, for which the name Pseudonocardia antimicrobica sp. nov. (type strain YIM 63235(T) =CCTCC AA 208080(T)=DSM 45303(T)) is proposed.

  2. Oligotrophy is Helpful for the Isolation of Bioactive Actinomycetes.

    PubMed

    Wang, Dong-Sheng; Xue, Quan-Hong; Ma, Yun-Yan; Wei, Xiao-Li; Chen, Jie; He, Fei

    2014-06-01

    It is necessary to develop new methods for the isolation of unknown actinomycetes from soils. To evaluate the effects of oligotrophic medium on the isolation of soil actinomycetes and develop a new isolation method, the Gause's synthetic medium was diluted to one tenth the recommended concentration in the present study. Soil dilution plate technique was used to isolate actinomycetes from the soil samples. Oligotrophy decreased actinomycete and streptomycete counts, as well as the number of antagonistic actinomycete species. Oligotrophy also decreased the number of actinomycete species in five samples. Some actinomycete species were cultured only on the oligotrophic medium, whereas other species could not be cultured. Oligotrophy decreased actinomycete counts more significantly for soils with organic matter content >40 g/kg. We used 16S rRNA sequence analysis to identify 22 actinomycete species that were only cultured on the oligotrophic medium. Oligotrophic medium was helpful for the isolation of Streptomyces spp., Micromonospora spp. and Streptosporangium spp. Slightly more than 80 % of the identified actinomycete species were biologically active. Therefore, we could draw a conclusion that oligotrophic medium could be helpful for the discovery of new antibiotic producers and the exploitation and utilization of new, biologically active compounds.

  3. Rare actinomycetes Nocardia caishijiensis and Pseudonocardia carboxydivorans as endophytes, their bioactivity and metabolites evaluation.

    PubMed

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida; Kulik, Andreas; Grond, Stephanie

    2016-04-01

    Two strains identified as Nocardia caishijiensis (SORS 64b) and Pseudonocardia carboxydivorans (AGLS 2) were isolated as endophytes from Sonchus oleraceus and Ageratum conyzoides respectively. The analysis of their extracts revealed them to be strongly bioactive. The N. caishijiensis extract gave an LC50 of 570 μg/ml(-1) in the brine shrimp cytotoxicity assay and an EC50 of 0.552 μg/ml(-1) in the DPPH antioxidant assay. Antimicrobial activity was observed against Methicillin resistant Staphlococcus aureus (MRSA) and Escherichia coli ATCC 25922 (14 mm), Klebsiella pneumoniae ATCC 706003 (13 mm), S. aureus ATCC 25923 (11 mm) and Candida tropicalis (20 mm). For the extract of P. carboxydivorans the EC50 was 0.670 μg/ml(-1) and it was observed to be more bioactive against Bacillus subtilis DSM 10 ATCC 6051 (21 mm), C. tropicalis (20 mm), S. aureus ATCC 25923 (17 mm), MRSA (17 mm), E. coli K12 (W1130) (16 mm) and Chlorella vulgaris (10 mm). The genotoxicity testing revealed a 20 mm zone of inhibition against the polA mutant strain E. coli K-12 AB 3027 suggesting damage to the DNA and polA genes. The TLC and bioautography screening revealed a diversity of active bands of medium polar and nonpolar compounds. Metabolite analysis by HPLC-DAD via UV/vis spectral screening suggested the possibility of stenothricin and bagremycin A in the mycelium extract of N. caishijiensis respectively. In the broth and mycelium extract of P. carboxydivorans borrelidin was suggested along with α-pyrone. The HPLC-MS revealed bioactive long chained amide derivatives such as 7-Octadecenamide, 9, 12 octadecandienamide. This study reports the rare actinomycetes N. caishijiensis and P. carboxydivorans as endophytes and evaluates their bioactive metabolites.

  4. Microbispora bryophytorum sp. nov., an actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Zhang, Yuejing; Liu, Chongxi; Wang, Haiyan; Zhao, Junwei; Li, Lianjie; Zhang, Zhongwen; Wang, Xiangjing; Xiang, Wensheng

    2015-04-01

    A novel endophytic actinomycete, designated strain NEAU-TX2-2(T), was isolated from moss and characterized using a polyphasic approach. The isolate was found to have morphological characteristics typical of the genus Microbispora . The isolate formed longitudinally paired spores on the tips of short sporophores that branched from aerial hyphae. Analysis of the 16S rRNA gene sequence supported the assignment of the novel strain to the genus Microbispora , and strain NEAU-TX2-2(T) exhibited 99.08 and 98.62% gene sequence similarities to Microbispora amethystogenes JCM 3021(T) and Microbispora rosea subsp. rosea JCM 3006(T), respectively. However two tree-making algorithms supported the position that strain NEAU-TX2-2(T) formed a distinct clade with M. rosea subsp. rosea JCM 3006(T). A low level of DNA-DNA relatedness allowed the isolate to be differentiated from M. amethystogenes JCM 3021(T) and M. rosea subsp. rosea JCM 3006(T). Moreover, strain NEAU-TX2-2(T) could also be distinguished from its closest phylogenetic relatives by morphological and physiological characteristics. Therefore, it is proposed that strain NEAU-TX2-2(T) represents a novel species of the genus Microbispora for which the name Microbispora bryophytorum sp. nov. is proposed. The type strain is NEAU-TX2-2(T) ( = CGMCC 4.7138(T) = DSM 46710(T)).

  5. Metabolites of Streptomyces sp., an endophytic actinomycete from Alpinia oxyphylla.

    PubMed

    Zhou, Hao; Yang, Yabin; Peng, Tianfeng; Li, Wei; Zhao, Lixing; Xu, Lihua; Ding, Zhongtao

    2014-01-01

    Four known compounds had been isolated from the Streptomyces sp. YIM66017, and their structures were elucidated by spectral analysis as 2,6-dimethoxy terephthalic acid (1), yangjinhualine A (2), α-hydroxyacetovanillone (3) and cyclo(Gly-Trp) (4). Compound 1 was isolated from natural resources for the first time, and compounds 2-4 were isolated from streptomycetes for the first time. The 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity assays showed that 1 showed higher activity than rutin with IC50 of 4.61 μg/mL and 2 showed the activity with IC50 of 57.12 μg/mL.

  6. Antimicrobial potential of Actinomycetes species isolated from marine environment.

    PubMed

    Valli, S; Suvathi, Sugasini S; Aysha, O S; Nirmala, P; Vinoth, Kumar P; Reena, A

    2012-06-01

    To evaluate the antimicrobial activity of Actinomycetes species isolated from marine environment. Twenty one strains of Actinomycetes were isolated from samples of Royapuram, Muttukadu, Mahabalipuram sea shores and Adyar estuary. Preliminary screening was done using cross-streak method against two gram-positive and eight gram-negative bacteria. The most potent strains C11 and C12 were selected from which antibacterial substances were extracted. The antibacterial activities of the extracts were performed using Kirby-Bauer disc diffusion method. Molecular identification of those isolates was done. All those twenty one isolates were active against at least one of the test organisms. Morphological characters were recorded. C11 showed activity against Staphylococcus species (13.0±0.5 mm), Vibrio harveyi (11.0±0.2 mm), Pseudomonas species (12.0±0.3 mm). C12 showed activity against Staphylococcus species (16.0±0.4 mm), Bacillus subtilis (11.0±0.2 mm), Vibrio harveyi (9.0±0.1 mm), Pseudomonas species (10.0±0.2 mm). 16S rRNA pattern strongly suggested that C11 and C12 strains were Streptomyces species. The results of the present investigation reveal that the marine Actinomycetes from coastal environment are the potent source of novel antibiotics. Isolation, characterization and study of Actinomycetes can be useful in discovery of novel species of Actinomycetes.

  7. Endophytic actinomycetes from spontaneous plants of Algerian Sahara: indole-3-acetic acid production and tomato plants growth promoting activity.

    PubMed

    Goudjal, Yacine; Toumatia, Omrane; Sabaou, Nasserdine; Barakate, Mustapha; Mathieu, Florence; Zitouni, Abdelghani

    2013-10-01

    Twenty-seven endophytic actinomycete strains were isolated from five spontaneous plants well adapted to the poor sandy soil and arid climatic conditions of the Algerian Sahara. Morphological and chemotaxonomical analysis indicated that twenty-two isolates belonged to the Streptomyces genus and the remaining five were non-Streptomyces. All endophytic strains were screened for their ability to produce indole-3-acetic acid (IAA) in vitro on a chemically defined medium. Eighteen strains were able to produce IAA and the maximum production occurred with the Streptomyces sp. PT2 strain. The IAA produced was further extracted, partially purified and confirmed by thin layer chromatography (TLC) analysis. The 16S rDNA sequence analysis and phylogenetic studies indicated that strain PT2 was closely related to Streptomyces enissocaecilis NRRL B 16365(T), Streptomyces rochei NBRC 12908(T) and Streptomyces plicatus NBRC 13071(T), with 99.52 % similarity. The production of IAA was affected by cultural conditions such as temperature, pH, incubation period and L-tryptophan concentration. The highest level of IAA production (127 μg/ml) was obtained by cultivating the Streptomyces sp. PT2 strain in yeast extract-tryptone broth supplemented with 5 mg L-tryptophan/ml at pH 7 and incubated on a rotary shaker (200 rpm) at 30 °C for 5 days. Twenty-four-hour treatment of tomato cv. Marmande seeds with the supernatant culture of Streptomyces sp. PT2 that contained the crude IAA showed the maximum effect in promoting seed germination and root elongation.

  8. Cultivation-independent population analysis of bacterial endophytes in three potato varieties based on eubacterial and Actinomycetes-specific PCR of 16S rRNA genes.

    PubMed

    Sessitsch, Angela; Reiter, Birgit; Pfeifer, Ulrike; Wilhelm, Eva

    2002-01-01

    Abstract Endophytic bacteria are ubiquitous in most plants and colonise plants without exhibiting pathogenicity. Studies on the diversity of bacterial endophytes have been mainly approached by characterisation of isolates obtained from internal tissues. Despite the broad application of culture-independent techniques for the analysis of microbial communities in a wide range of natural habitats, little information is available on the species diversity of endophytes. In this study, microbial communities inhabiting stems, roots and tubers of three potato varieties were analysed by 16S rRNA-based techniques such as terminal restriction fragment length polymorphism analysis, denaturing gradient gel electrophoresis as well as 16S rDNA cloning and sequencing. Two individual plant experiments were conducted. In the first experiment plants suffered from light deficiency, whereas healthy and robust plants were obtained in the second experiment. Plants obtained from both experiments showed comparable endophytic populations, but healthy potato plants possessed a significantly higher diversity of endophytes than stressed plants. In addition, plant tissue and variety specific endophytes were detected. Sequence analysis of 16S rRNA genes indicated that a broad phylogenetic spectrum of bacteria is able to colonise plants internally including alpha-, beta-, and gamma-Proteobacteria, high-GC Gram-positives, microbes belonging to the Flexibacter/Cytophaga/Bacteroides group and Planctomycetales. Group-specific analysis of Actinomycetes indicated a higher abundance and diversity of Streptomyces scabiei-related species in the variety Mehlige Mühlviertler, which is known for its resistance against potato common scab caused by S. scabiei.

  9. Diversity and isolation of rare actinomycetes: an overview.

    PubMed

    Tiwari, Kavita; Gupta, Rajinder K

    2013-08-01

    A renewed interest in the development of new antimicrobial agents is urgently needed to combat the increasing number of antibiotic-resistant strains of pathogenic microorganisms. Actinomycetes continue to be the mainstream supplier of antibiotics used in industry. The likelihood of discovering a new compound with novel chemical structure can be increased with intensive efforts in isolating and screening of rare genera of microorganisms to include in natural-product-screening collections. An unexpected variety of rare actinomycetes is now being isolated worldwide from previously uninvestigated diverse natural habitats, using different selective isolation methods. These isolation efforts include methods to enhance growth (enrichment) of rare actinomycetes, and eliminate unwanted microorganisms (pretreatment). To speed up the strain isolation process, knowledge about the distribution of such unexploited groups of microorganisms must also be augmented. This is a summary of using these microorganisms as new potential biological resources, and a review of almost all of the selective isolation methods, including pretreatment and enrichment techniques that have been developed to date for the isolation of rare actinomycetes.

  10. Isolation and screening of endophytes from the rhizomes of some Zingiberaceae plants for L-asparaginase production.

    PubMed

    Krishnapura, Prajna Rao; Belur, Prasanna D

    2016-01-01

    Endophytes are described as microorganisms that colonize the internal tissues of healthy plants without causing any disease. Endophytes isolated from medicinal plants have been attracting considerable attention due to their high biodiversity and their predicted potential to produce a plethora of novel compounds. In this study, an attempt was made to isolate endophytes from rhizomes of five medicinal plants of Zingiberaceae family, and to screen the endophytes for L-asparaginase activity. In total, 50 endophytes (14 bacteria, 22 actinomycetes, and 14 fungi) were isolated from Alpinia galanga, Curcuma amada, Curcuma longa, Hedychium coronarium, and Zingiber officinale; of these, 31 endophytes evidenced positive for L-asparaginase production. All the L-asparaginase-positive isolates showed L-asparaginase activity in the range of 54.17-155.93 U/mL in unoptimized medium. An endophytic fungus isolated from Curcuma amada, identified as Talaromyces pinophilus, was used for further experiments involving studies on the effect of certain nutritional and nonnutritional factors on L-asparaginase production in submerged fermentation. Talaromyces pinophilus initially gave an enzyme activity of 108.95 U/mL, but gradually reduced to 80 U/mL due to strain degeneration. Perhaps this is the first report ever on the production of L-asparaginase from endophytes isolated from medicinal plants of Zingiberaceae family.

  11. Comparative analysis of chemical constituents, antimicrobial and antioxidant activities of ethylacetate extracts of Polygonum cuspidatum and its endophytic actinomycete, Streptomyces sp. A0916.

    PubMed

    Wang, Lei; Qiu, Peng; Long, Xiu-Feng; Zhang, Shuai; Zeng, Zhi-Gang; Tian, Yong-Qiang

    2016-02-01

    The present study investigated the chemical composition of ethylacetate extracts from an endophytic actinomycete Streptomyces sp. A0916 and its host Polygonum cuspidatum. A comparative analysis of the antimicrobial and antioxidant properties of the extracts was also conducted. 32 compounds of P. cuspidatum and 23 compounds of Streptomyces sp. A0916 were isolated and identified by GC/MS. Antimicrobial activities of the extracts were evaluated using eight microbial strains (3 Gram-positive bacteria, 3 Gram-negative bacteria, and 2 fungi). The Streptomyces sp. A0916 extracts showed a wide range of antimicrobial activities and presented greater antimicrobial effectiveness than the P. cuspidatum extracts. The minimum inhibitory concentration (MIC) of Streptomyces sp. A0916 extracts against the ampicillin-resistant strain Enterococcus faecium SIIA843 was 32 μg·mL(-1). Furthermore, the extracts had greater antimicrobial effect against Gram-positive bacteria than Gram-negative bacteria. Finally, the antioxidant activity of the Streptomyces sp. A0916 extracts was equal to that of the P. cuspidatum extracts. In conclusion, our results suggest that the endophytic actinomycetes of the medicinal plants are an important source of bioactive substances.

  12. Antibiotics production by an actinomycete isolated from the termite gut.

    PubMed

    Matsui, Toru; Tanaka, Junichi; Namihira, Tomoyuki; Shinzato, Naoya

    2012-12-01

    As well as the search for new antibiotics, a new resource or strains for the known antibiotics is also important. Microbial symbionts in the gut of termites could be regarded as one of the feasible resource for such purpose. In this study, antibiotic-producing actinomycetes were screened from symbionts of the termite gut. 16SrRNA sequence analysis for the 10 isolates revealed that they belong to actinomycetes such as Streptomyces sp., Kitasatospora sp., and Mycobacterium sp. A culture broth from one of the isolate, namely strain CA1, belonging to the genera Streptomyces exhibited antagonistic activity against actinomycetes (Micrococcus spp.), gram-positive bacteria (Bacillus spp.), and yeast (Candida spp.). The structures of 2 compounds isolated from the culture broth of the strain CA1 were identified as those of actinomycin X2 and its analog, D. This study is the first to report that some symbionts of the termite gut are antibiotic-producing actinomycetes, and suggest that the termite gut is a feasible resource for bioprospecting.

  13. Isolation and evaluation of proteolytic actinomycete isolates as novel inducers of pearl millet downy mildew disease protection

    PubMed Central

    Jogaiah, Sudisha; Kurjogi, Mahantesh; Govind, Sharathchandra Ramasandra; Huntrike, Shekar Shetty; Basappa, Vedamurthy Ankala; Tran, Lam-Son Phan

    2016-01-01

    Native endophytic actinomycetes isolated from pearl millet roots were examined for their efficacy to protect pearl millet against downy mildew. Nineteen of 39 isolates were found to be proteolytic, of which 7 strains could directly suppress the sporangium formation of Sclerospora graminicola, the pearl millet downy mildew pathogen. Thus, mycelial suspensions containing either spores or cell-free extract of these 7 isolates were used for seed-coating and -soaking treatments to test for their induction of downy mildew resistance. Results indicated that seed-coating overall provided better protection to downy mildew than seed-soaking. In both treatments, the tested isolates demonstrated differential abilities in downy mildew disease protection, with Streptomyces griseus SJ_UOM-07-09 and Streptosporangium roseum SJ_UOM-18-09 showing the highest protection rates. Additionally, the levels of disease protection conferred by the actinomycetes were just slightly lower than that of the systemic fungicide Apron, suggesting their effectiveness. Further studies revealed that the more rapid root colonization by SJ_UOM-18-09 resulted in faster and higher induced resistance in comparison with SJ_UOM-07-09 under greenhouse conditions, indicating that SJ_UOM-18-09 was superior than SJ_UOM-07-09 in inducing resistance. Results from this study provide comprehensive information on biocontrol functions of SJ_UOM- 18-09 with great potential to control downy mildew disease in pearl millet. PMID:27499196

  14. Isolation and evaluation of proteolytic actinomycete isolates as novel inducers of pearl millet downy mildew disease protection.

    PubMed

    Jogaiah, Sudisha; Kurjogi, Mahantesh; Govind, Sharathchandra Ramasandra; Huntrike, Shekar Shetty; Basappa, Vedamurthy Ankala; Tran, Lam-Son Phan

    2016-08-08

    Native endophytic actinomycetes isolated from pearl millet roots were examined for their efficacy to protect pearl millet against downy mildew. Nineteen of 39 isolates were found to be proteolytic, of which 7 strains could directly suppress the sporangium formation of Sclerospora graminicola, the pearl millet downy mildew pathogen. Thus, mycelial suspensions containing either spores or cell-free extract of these 7 isolates were used for seed-coating and -soaking treatments to test for their induction of downy mildew resistance. Results indicated that seed-coating overall provided better protection to downy mildew than seed-soaking. In both treatments, the tested isolates demonstrated differential abilities in downy mildew disease protection, with Streptomyces griseus SJ_UOM-07-09 and Streptosporangium roseum SJ_UOM-18-09 showing the highest protection rates. Additionally, the levels of disease protection conferred by the actinomycetes were just slightly lower than that of the systemic fungicide Apron, suggesting their effectiveness. Further studies revealed that the more rapid root colonization by SJ_UOM-18-09 resulted in faster and higher induced resistance in comparison with SJ_UOM-07-09 under greenhouse conditions, indicating that SJ_UOM-18-09 was superior than SJ_UOM-07-09 in inducing resistance. Results from this study provide comprehensive information on biocontrol functions of SJ_UOM- 18-09 with great potential to control downy mildew disease in pearl millet.

  15. [Isolation and antimicrobial activities of actinomycetes from vermicompost].

    PubMed

    Wang, Xue-jun; Yan, Shuang-lin; Min, Chang-li; Yang, Yan

    2015-02-01

    In this paper, actinomycetes were isolated from vermicompost by tablet coating method. Antimicrobial activities of actinomycetes were measured by the agar block method. Strains with high activity were identified based on morphology and biochemical characteristics, as well as 16S rDNA gene sequence analysis. The results showed that 26 strains of actinomycetes were isolated, 16 of them had antimicrobial activities to the test strains which accounts for 61.54% of all strains. Among the 16 strains, the strain QYF12 and QYF22 had higher antimicrobial activity to Micrococcus luteus, with a formed inhibition zone of 27 mm and 31 mm, respectively. While the strain QYF26 had higher antimicrobial activity to Bacillus subtilis, and the inhibition zone diameter was 21 mm. Based on the identification of strains with high activity, the strain QYF12 was identified as Streptomyces chartreusis, the strain QYF22 was S. ossamyceticus and the strain QYF26 was S. gancidicus. This study provided a theoretical basis for further separate antibacterial product used for biological control.

  16. Geosmin, an Earthy-Smelling Substance Isolated from Actinomycetes

    PubMed Central

    Gerber, N. N.; Lechevalier, H. A.

    1965-01-01

    Geosmin, an earthy-smelling substance, has been isolated from several actinomycetes. Production of 1 mg per liter of whole broth was obtained from Streptomyces griseus LP-16. After preliminary separations, pure geosmin was isolated in milligram amounts by gas chromatography. Geosmin is a neutral oil, with an approximate boiling point of 270 C, which contains carbon and hydrogen, but no nitrogen. It undergoes a reaction with acid to give odorless argosmin, a neutral oil, with an approximate boiling point of 230 C, which contains only carbon and hydrogen. Specific rotation and ultraviolet- and infrared-absorbtion spectra were determined for both. PMID:5866039

  17. Saccharothrix hoggarensis sp. nov., an actinomycete isolated from Saharan soil.

    PubMed

    Boubetra, Dalila; Zitouni, Abdelghani; Bouras, Noureddine; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-02-01

    An actinomycete, designated SA181(T), was isolated from Saharan soil in the Hoggar region (south Algeria) and was characterized taxonomically by using a polyphasic approach. The morphological and chemotaxonomic characteristics of the isolate were consistent with the genus Saccharothrix, and 16S rRNA gene sequence analysis confirmed that strain SA181(T) was a novel member of the genus Saccharothrix. DNA-DNA hybridization values between strain SA181(T) and its closest phylogenetic neighbours, the type strains of Saccharothrix longispora, Saccharothrix texasensis and Saccharothrix xinjiangensis, were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the isolate represents a novel species of the genus Saccharothrix, for which the name Saccharothrix hoggarensis sp. nov. is proposed, with the type strain SA181(T) ( = DSM 45457(T)  = CCUG 60214(T)).

  18. Actinoalloteichus hoggarensis sp. nov., an actinomycete isolated from Saharan soil.

    PubMed

    Boudjelal, Farida; Zitouni, Abdelghani; Bouras, Noureddine; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-06-01

    A moderately halophilic actinomycete strain, designated AH97T, was isolated from Saharan soil in the Hoggar region (south Algeria) and was subjected to polyphasic taxonomic characterization. The morphological and chemotaxonomic characteristics of the strain were consistent with those of the genus Actinoalloteichus. Results of 16S rRNA gene sequence comparison revealed that strain AH97T shared the highest degree of 16S rRNA gene sequence similarity with Actinoalloteichus hymeniacidonis DSM 45092T (99.3 %) and Actinoalloteichus nanshanensis DSM 45655T (98.7 %). However, DNA-DNA hybridization studies showed only 26.5 % relatedness with A. hymeniacidonis DSM 45092T and 28.0 % with A. nanshanensis DSM 45655T. The genotypic and phenotypic data showed that strain AH97T represents a novel species of the genus Actinoalloteichus, for which the name Actinoalloteichus hoggarensis sp. nov. is proposed, with AH97T ( = DSM 45943T = CECT 8639T) as the type strain.

  19. Saccharothrix tamanrassetensis sp. nov., an actinomycete isolated from Saharan soil.

    PubMed

    Boubetra, Dalila; Zitouni, Abdelghani; Bouras, Noureddine; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2015-04-01

    Actinomycete strain SA198(T), isolated from a Saharan soil sample of Algeria, was characterized taxonomically by using a polyphasic approach. Chemotaxonomic and morphological characteristics observed suggested that it was a member of the genus Saccharothrix . The 16S rRNA gene sequence analysis confirmed that strain SA198(T) was a member of the genus Saccharothrix and showed a similarity level ranging between 97.5 and 98.9% within species of the genus Saccharothrix , Saccharothrix australiensis being the most closely related. However, DNA-DNA hybridization values between strain SA198(T) and its closest phylogenetic neighbours, the type strains of S. australiensis , Saccharothrix xinjiangensis , Saccharothrix algeriensis and Saccharothrix espanaensis , were clearly below the 70% threshold. Based upon genotypic and phenotypic differences from other members of the genus, a novel species, Saccharothrix tamanrassetensis sp. nov., is proposed, with SA198(T) ( = DSM 45947(T) = CECT 8640(T)) as the type strain.

  20. A novel method to scale up fungal endophyte isolations

    USDA-ARS?s Scientific Manuscript database

    Estimations of species diversity are influenced by sampling intensity which in turn is influenced by methodology. For fungal endophyte diversity studies, the methodology includes surface-sterilization prior to isolation of endophytes. Surface-sterilization is an essential component of fungal endophy...

  1. [Phylogenetic diversity of the culturable rare actinomycetes in marine sponge Hymeniacidon perlevis by improved isolation media].

    PubMed

    Xin, Yanjuan; Wu, Peichun; Deng, Maicun; Zhang, Wei

    2009-07-01

    Based on the molecular diversity information, seven actinomycete-selective culture media and isolation conditions were modified to isolate and cultivate diverse rare actinomycetes from Hymeniacidon perlevis. Modified, selective cultivation and enrichment media were used, with the addition of an elemental solution of simulating the elemental composition of marine sponge H. perlevis. Restriction Fragment Length Polymorphism (RFLP) analysis of 16S rDNA sequence was used to reveal the diversity of culturable rare actinomycetes. A total of 59 actinomycete strains were isolated from the marine sponge H. perlevis. A total of 27 representative actinomycetes were selected according to their morphological feature, color and pigments. They gave 15 different RFLP patterns after digesting their PCR products of 16s rDNA with Hha I. The results showed that these isolates belonged to 10 genera: Streptomyces, Nocardiopsis, Micromonospora, Cellulosimicrobium, Gordonia, Nocardia, Prauseria, Pseudonocardia , Saccharomonospora and Microbacterium. The modified isolation media and selective cultivation procedures are highly effective in the recovery of culturable actinomycetes from the marine sponge H. perlevis, resulting in the highest diversity of culturable rare actinomycetes from any sponges.

  2. Terrabacter lapilli sp. nov., an actinomycete isolated from stone.

    PubMed

    Lee, Jeong-Eon; Seo, Jae Pyo; Lee, Dong Wan; Ko, Young-Hwan; Lee, Soon Dong

    2008-05-01

    A novel actinomycete, designated strain LR-26T, was isolated from a small stone collected from an agricultural field in Jeju, Republic of Korea. Cells of the organism were strictly aerobic, Gram-positive, non-motile, short rods. Colonies were bright yellow, circular, smooth and translucent. The organism was characterized chemotaxonomically as having ll-diaminopimelic acid in the cell wall, MK-8(H4) as major menaquinone, a polar lipid profile including diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and unknown phospholipids, iso-C15 : 0 as the predominant fatty acid and a DNA G+C content of 72.6 mol%. Comparative 16S rRNA gene sequence analysis showed that the organism was related to the genera Intrasporangium, Terracoccus and Terrabacter within the family Intrasporangiaceae. The closest phylogenetic relatives of strain LR-26T were the type strains of Terrabacter terrae (99.4 % 16S rRNA gene sequence similarity), Terrabacter aerolatus (99.3 %) and Terrabacter tumescens (99.3 %). DNA-DNA hybridization experiments showed that strain LR-26T shared low levels of DNA-DNA relatedness with Terrabacter terrae LMG 22921T (17.6 and 22.8 % from reciprocal experiments) and with Terrabacter tumescens IMSNU 21313T (27.6 and 34.9 %). The phenotypic data and low levels of DNA-DNA relatedness readily distinguished strain LR-26T from the type strains of recognized species of the genus Terrabacter, and showed that it therefore represents a novel species. The name Terrabacter lapilli sp. nov. is proposed for this novel actinomycete. The type strain is LR-26T (=JBRI 2002T =KCTC 19199T =DSM 18583T).

  3. Isolation and Characterization of Actinomycete Antagonists of a Fungal Root Pathogen †

    PubMed Central

    Crawford, Don L.; Lynch, James M.; Whipps, John M.; Ousley, Margaret A.

    1993-01-01

    By use of selective media, 267 actinomycete strains were isolated from four rhizosphere-associated and four non-rhizosphere-associated British soils. Organic media with low nutrient concentrations were found to be best for isolating diverse actinomycetes while avoiding contamination and overgrowth of isolation media by eubacteria and fungi. While all isolates grew well at pHs 6.5 to 8.0, a few were unable to grow at pH 6.0 and a significant number failed to grow at pH 5.5. Eighty-two selected isolates were screened for in vitro antagonism towards Pythium ultimum by use of a Difco cornmeal agar assay procedure. Five isolates were very strong antagonists of the fungus, four were strong antagonists, and ten others were weakly antagonistic. The remaining isolates showed no antagonism by this assay. Additional studies showed that several of the P. ultimum antagonists also strongly inhibited growth of other root-pathogenic fungi. Twelve isolates showing antifungal activity in the in vitro assay were also tested for their effects on the germination and short-term growth of lettuce plants in glasshouse pot studies in the absence of pathogens. None of the actinomycetes prevented seed germination, although half of the isolates retarded seed germination and outgrowth of the plants by 1 to 3 days. During 18-day growth experiments, biomass yields of some actinomycete-inoculated plants were reduced in comparison with untreated control plants, although all plants appeared healthy and well rooted. None of the actinomycetes significantly enhanced plant growth over these short-term experiments. For some, but not all, actinomycetes, some correlations between delayed seed germination and reduced 18-day plant biomass yields were seen. For others, plant biomass yields were not reduced despite an actinomycete-associated delay in seed germination and plant outgrowth. Preliminary glasshouse experiments indicated that some of the actinomycetes protect germinating lettuce seeds against

  4. Diversity of actinomycetes isolated from subseafloor sediments after prolonged low-temperature storage.

    PubMed

    Ulanova, Dana; Goo, Kian-Sim

    2015-05-01

    Subseafloor sediments present an untapped source of novel bacterial species with industrially important bioactivities. Subseafloor core samples collected during the Integrated Ocean Drilling Program Expeditions 315, 316, and 331 and stored in Kochi Core Center at -80 °C for 1 to 4 years were used for cultivation-based study of viable actinomycetes. In total, more than 100 actinomycete-like colonies were isolated from two deep-frozen subseafloor sediment samples. Isolated actinomycetes showed close similarity to known Actinotalea, Dietzia, Gordonia, Isoptericola, Microbacterium, Nocardia, Rhodococcus, Pseudonocardia, Streptomyces, and Tsukamurella species and were halotolerant. Bioactivity assays revealed that two of the isolates were producing potent antibacterial compound(s) and one isolate was having antifungal activity. Our study demonstrated that deep-frozen subseafloor core samples could be a potential source of viable actinomycetes, which may be used in drug discovery.

  5. Isolation and evaluation of endophytic Streptomyces endus OsiSh-2 with potential application for biocontrol of rice blast disease.

    PubMed

    Xu, Ting; Li, Yan; Zeng, Xiadong; Yang, Xiaolu; Yang, Yuanzhu; Yuan, Shanshan; Hu, Xiaochun; Zeng, Jiarui; Wang, Zhenzhen; Liu, Qian; Liu, Yuqing; Liao, Hongdong; Tong, Chunyi; Liu, Xuanming; Zhu, Yonghua

    2017-03-01

    Biocontrol is a promising strategy in the control of rice blast disease. In the present study, we isolated and characterized a novel antagonist to the pathogen Magnaporthe oryzae from rice endophytic actinomycetes. Out of 482 endophytic actinomycetes isolated from rice blast infected and healthy rice, Streptomyces endus OsiSh-2 exhibited remarkable in vitro antagonistic activity. Scanning electron microscopy observations of M. oryzae treated by OsiSh-2 revealed significant morphological alterations in hyphae. In 2-year field tests, the spraying of OsiSh-2 spore solution (10(7)  spores mL(-1) ) is capable of reducing rice blast disease severity by 59.64%. In addition, a fermentation broth of OsiSh-2 and its cell-free filtrates could inhibit the growth of M. oryzae, suggesting the presence of active enzymes and secondary metabolites. OsiSh-2 tested positive for polyketide synthase-I and nonribosomal peptide synthetase genes and can produce cellulase, protease, gelatinase, siderophore, indole-3-acetic acid and 1-amino-cyclopropane-1-carboxylate deaminase. A preliminary separation indicated that the methanol extract of OsiSh-2 could suppress the growth of pathogens. The major active component was identified as nigericin. Endophytic S. endus OsiSh-2 has potential as a biocontrol agent against rice blast in agriculture. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  6. Actinomycetes with antimicrobial activity isolated from paper wasp (Hymenoptera: Vespidae: Polistinae) nests.

    PubMed

    Madden, Anne A; Grassetti, Andrew; Soriano, Jonathan-Andrew N; Starks, Philip T

    2013-08-01

    Actinomycetes-a group of antimicrobial producing bacteria-have been successfully cultured and characterized from the nest material of diverse arthropods. Some are symbionts that produce antimicrobial chemicals found to protect nest brood and resources from pathogenic microbes. Others have no known fitness relationship with their associated insects, but have been found to produce antimicrobials in vitro. Consequently, insect nest material is being investigated as a new source of novel antimicrobial producing actinomycetes, which could be harnessed for therapeutic potential. To extend studies of actinomycete-insect associations beyond soil-substrate dwelling insects and wood boring excavators, we conducted a preliminary assessment of the actinomycetes within the nests of the paper wasp, Polistes dominulus (Christ). We found that actinomycetes were readily cultured from nest material across multiple invasive P. dominulus populations-including members of the genera Streptomyces, Micromonospora, and Actinoplanes. Thirty of these isolates were assayed for antimicrobial activity against the challenge bacteria Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Serratia marcescens, and Bacillus subtilis. Sixty percent of isolates inhibited the growth of at least one challenge strain. This study provides the first assessment of bacteria associated with nests of P. dominulus, and the first record of antimicrobial producing actinomycetes isolated from social wasps. We provide a new system to explore nest associated actinomycetes from a ubiquitous and cosmopolitan group of insects.

  7. Catellatospora aurea sp. nov., a novel actinomycete isolated from soil.

    PubMed

    Liu, Chongxi; Zhao, Junwei; Guan, Xuejiao; Li, Lianjie; Li, Wenbin; Wang, Xiangjing; Xiang, Wensheng

    2014-12-01

    A novel actinomycete, designated strain NEAU-SH16(T) was isolated from a soil sample collected from the riverbank of Wusong river in Shanghai and characterized using a polyphasic approach. Morphological and chemotaxonomic properties of strain NEAU-SH16(T) were consistent with the description of the genus Catellatospora. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that strain NEAU-SH16(T) formed a distinct branch with the closest relatives Catellatospora coxensis DSM 44901(T) (99.2 %) and Catellatospora citrea DSM 44097(T) (99.0 %), an association that was supported by a bootstrap value of 78 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. However, the low level of DNA-DNA relatedness and some different physiological characteristics allowed the strain to be distinguished from its closest relatives. Thus, it is proposed that strain NEAU-SH16(T) represents a novel Catellatospora species. Catellatospora aurea sp. nov. The type strain of Catellatospora aurea is NEAU-SH16(T) (=CGMCC 4.7147(T) = DSM 46719(T)).

  8. Isolation and characterization of medically important aerobic actinomycetes in soil of iran (2006 - 2007).

    PubMed

    Aghamirian, Mohammad Reza; Ghiasian, Seyed Amir

    2009-01-01

    The aerobic actinomycetes are a large group of soil-inhabiting bacteria that occur worldwide. Some of them are the main cause of two important diseases, nocardiosis and actinomycetoma. To identify the prevalence and geographic distribution of aerobic actinomycetes in soil of Qazvin province, a study was carried out during 2006-2007. In this study, the incidence and diversity of medically important aerobic actinomycetes was determined in 300 soil samples of different parts of Qazvin. The suspensions of superficial soil samples were prepared by adding of normal saline, streptomycin and chloramphenicol and the supernatants were cultured on brain-heart infusion agar and Sabouraud's dextrose agar contain cycloheximide. The isolated microorganisms were examined by Gram and acid-fast stains and were identified biochemically and morphologically. Of 96 aerobic actinomycetes isolates identified, Actinomadura madurae and Streptomyces somaliensis were the most frequently isolated species each representing 19.8% of isolates, followed by Nocardia asteroides (15.6%), N. otitidiscaviarum (9.4%), N. brasiliensis (7.3%), A. peletieri, S. griseus, and Nocardia spp. (each 5.2%), and N. transvalensis, Nocardiopsis dassonvillei, Actinomadura spp. and Streptomyces spp. (each 3.1%). To the best of our knowledge, this is the first report on epidemiological investigation of medically important aerobic actinomycetes in soil samples from Iran. In recent years, mycetoma and nocardiosis have been increasingly reported in Iran. The results showed that medically important actinomycetes occur in the environment of Iran and soil could be potential source of actinomycotic infections.

  9. Isolation strategies of marine-derived actinomycetes from sponge and sediment samples.

    PubMed

    Hameş-Kocabaş, E Esin; Uzel, Ataç

    2012-03-01

    During the last two decades, discoveries of new members of actinomycetes and novel metabolites from marine environments have drawn attention to such environments, such as sediment and sponge. For the successful isolation of actinomycetes from marine environments, many factors including the use of enrichment and pre-treatment techniques, and the selection of growth media and antibiotic supplements should be taken into account. High-throughput cultivation is an innovative technique that mimics nature, eliminates undesired, fast-growing bacteria and creates suitable conditions for rare, slow-growing actinomycetes. This review comprehensively evaluates the traditional and innovative techniques and strategies used for the isolation of actinomycetes from marine sponge and sediment samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Isolation and identification of actinomycetes for production of novel extracellular glutaminase free L-asparaginase.

    PubMed

    Saxena, Akansha; Upadhyay, Ramraj; Kango, Naveen

    2015-12-01

    Over the recent years glutaminase free L-asparaginase has gained more importance due to better therapeutic properties for treatment of acute lymphoblastic leukemia. Actinomycetes are known for L-asparaginase activity. In the current study, 80 actinomycetes were isolated from various soil habitats by serial dilution technique. Presence of L-asparaginase was investigated in a total of 240 actinomycetes by tubed agar method using modified M-9 medium. A total of 165 actinomycetes were found positive for L-asparaginase activity. Among these, 57 actinomycetes producing larger zones of L-asparagine hydrolysis were further screened for their capacity to produce glutaminase-free L-asparaginase. Four L-glutaminase-free actinomycetes were found to be potential L-asparaginase producers. These actinomycetes were identified as Streptomyces cyaneus (SAP 1287, CFS 1560), S. exfoliates (CFS 1557) and S. phaeochromogenes (GS 1573) on the basis of morphological and biochemical identification studies. Maximum L-asparaginase activity (19.2 Uml(-1)) was observed in culture filtrate of S. phaeochromogenes under submerged fermentation. Results indicate that S. phaeochromogenes could be a potential source of glutaminase free L-asparaginase for commercial purpose. To the best of our knowledge, this is the first report on production of glutaminase free L-asparaginase from S. cyaneus, S. exfoliatus and S. phaeochromogenes.

  11. Isolation and characterization of bacterial endophytes of Chelidonium majus L.

    PubMed

    Goryluk, Agata; Rekosz-Burlaga, Hanna; Błaszczyk, Mieczysław

    2009-01-01

    The aim of this study was to isolate and identify endophytic bacteria from stems of Chelidonium majus L. (greater celandine) and to evaluate their antifungal properties. In total, 34 bacterial endophyte strains were isolated. The fungistatic effects of these bacteria on the growth of five moulds (Alternaria alternata, Chaetonium sp., Paecilomyces variotti, Byssochlamys fulva, Aureobasidium pullulans) and one species of black yeast (Exophiala mesophila) were tested. The majority of the bacterial isolates were found to inhibit the growth of fungi and those with the strongest antifungal properties were further characterized. Of the twelve isolates examined, 11 were species of Bacillus thuringiensis and one was Bacillus amyloliquefaciens.

  12. Streptomyces lopnurensis sp. nov., an actinomycete isolated from soil.

    PubMed

    Zheng, Bei; Han, Xiao-Xue; Xia, Zhan-Feng; Wan, Chuan-Xing; Zhang, Li-Li

    2014-12-01

    A novel actinomycete, designated strain TRM 49590(T), was isolated from a soil sample from Lop Nur in Xinjiang Province, China. Strain TRM 49590(T) was aerobic, Gram-staining-positive, with an optimum NaCl concentration for growth of 1.5 % (w/v) and an optimum temperature for growth of 28-37 °C. The aerial mycelium was sparse, cylindrical and smooth-surfaced with irregular branches on ISP medium 4. The whole-cell sugars of strain TRM 49590(T) were ribose and glucose. The diagnostic diamino acid contained ll-diaminopimelic acid. The predominant menaquinones were MK-9(H6) and MK-9(H8), with MK-9(H4) and MK-10(H6) present in smaller amounts. The major fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 62.2 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain TRM 49590(T) belongs to the genus Streptomyces with a sequence similarity of 97.16 % with the most closely related species Streptomyces sodiiphilus. Based on these observations, strain TRM 49590(T) is proposed to represent a novel species of the genus Streptomyces for which the name Streptomyces lopnurensis sp. nov. is suggested. The type strain is TRM 49590(T) ( = CCTCC AA 2013018(T) = NRRL B59109(T)).

  13. Actinomycetes for marine drug discovery isolated from mangrove soils and plants in China.

    PubMed

    Hong, Kui; Gao, An-Hui; Xie, Qing-Yi; Gao, Hao; Zhuang, Ling; Lin, Hai-Peng; Yu, Hai-Ping; Li, Jia; Yao, Xin-Sheng; Goodfellow, Michael; Ruan, Ji-Sheng

    2009-01-01

    The mangrove ecosystem is a largely unexplored source for actinomycetes with the potential to produce biologically active secondary metabolites. Consequently, we set out to isolate, characterize and screen actinomycetes from soil and plant material collected from eight mangrove sites in China. Over 2,000 actinomycetes were isolated and of these approximately 20%, 5%, and 10% inhibited the growth of Human Colon Tumor 116 cells, Candida albicans and Staphylococcus aureus, respectively, while 3% inhibited protein tyrosine phosphatase 1B (PTP1B), a protein related to diabetes. In addition, nine isolates inhibited aurora kinase A, an anti-cancer related protein, and three inhibited caspase 3, a protein related to neurodegenerative diseases. Representative bioactive isolates were characterized using genotypic and phenotypic procedures and classified to thirteen genera, notably to the genera Micromonospora and Streptomyces. Actinomycetes showing cytotoxic activity were assigned to seven genera whereas only Micromonospora and Streptomyces strains showed anti-PTP1B activity. We conclude that actinomycetes isolated from mangrove habitats are a potentially rich source for the discovery of anti-infection and anti-tumor compounds, and of agents for treating neurodegenerative diseases and diabetes.

  14. Actinomycetes for Marine Drug Discovery Isolated from Mangrove Soils and Plants in China

    PubMed Central

    Hong, Kui; Gao, An-Hui; Xie, Qing-Yi; Gao, Hao; Zhuang, Ling; Lin, Hai-Peng; Yu, Hai-Ping; Li, Jia; Yao, Xin-Sheng; Goodfellow, Michael; Ruan, Ji-Sheng

    2009-01-01

    The mangrove ecosystem is a largely unexplored source for actinomycetes with the potential to produce biologically active secondary metabolites. Consequently, we set out to isolate, characterize and screen actinomycetes from soil and plant material collected from eight mangrove sites in China. Over 2,000 actinomycetes were isolated and of these approximately 20%, 5%, and 10% inhibited the growth of Human Colon Tumor 116 cells, Candida albicans and Staphylococcus aureus, respectively, while 3% inhibited protein tyrosine phosphatase 1B (PTP1B), a protein related to diabetes. In addition, nine isolates inhibited aurora kinase A, an anti-cancer related protein, and three inhibited caspase 3, a protein related to neurodegenerative diseases. Representative bioactive isolates were characterized using genotypic and phenotypic procedures and classified to thirteen genera, notably to the genera Micromonospora and Streptomyces. Actinomycetes showing cytotoxic activity were assigned to seven genera whereas only Micromonospora and Streptomyces strains showed anti-PTP1B activity. We conclude that actinomycetes isolated from mangrove habitats are a potentially rich source for the discovery of anti-infection and anti-tumor compounds, and of agents for treating neurodegenerative diseases and diabetes. PMID:19370169

  15. Taxonomic and ecological studies of actinomycetes from Vietnam: isolation and genus-level diversity.

    PubMed

    Hop, Duong Van; Sakiyama, Yayoi; Binh, Chu Thi Thanh; Otoguro, Misa; Hang, Dinh Thuy; Miyadoh, Shinji; Luong, Dao Thi; Ando, Katsuhiko

    2011-09-01

    Actinomycetes were isolated from 109 soil and 93 leaf-litter samples collected at five sites in Vietnam between 2005 and 2008 using the rehydration-centrifugation (RC) method, sodium dodecyl sulfate-yeast extract dilution method, dry-heating method and oil-separation method in conjunction with humic acid-vitamin agar as an isolation medium. A total of 1882 strains were identified as Vietnamese (VN)-actinomycetes including 1080 (57%) streptomycetes (the genus Streptomyces isolates) and 802 (43%) non-streptomycetes. The 16S ribosomal RNA gene sequences of the VN-actinomycetes were analyzed using BLAST searches. The results showed that these isolates belonged to 53 genera distributed among 21 families. Approximately 90% of these strains were members of three families: Streptomycetaceae (1087 strains, 58%); Micromonosporaceae (516 strains, 27%); and Streptosporangiaceae (89 strains, 5%). Motile actinomycetes of the genera Actinoplanes, Kineosporia and Cryptosporangium, which have quite common morphological characteristics, were frequently isolated from leaf-litter samples using the RC method. It is possible that these three genera acquired common properties during a process of convergent evolution. By contrast, strains belonging to the suborder Streptosporangineae were exclusively isolated from soils. A comparison of the sampling sites revealed no significant difference in taxonomic diversity between these sites. Among the non-streptomycetes, 156 strains (19%) were considered as new taxa distributed into 21 genera belonging to 12 families. Interestingly, the isolation of actinomycetes from leaf-litter samples using the RC method proved to be the most efficient way to isolate new actinomycetes in Vietnam, especially the Micromonosporaceae species.

  16. Isolation, phylogenetic analysis and anti-infective activity screening of marine sponge-associated actinomycetes.

    PubMed

    Abdelmohsen, Usama Ramadan; Pimentel-Elardo, Sheila M; Hanora, Amro; Radwan, Mona; Abou-El-Ela, Soad H; Ahmed, Safwat; Hentschel, Ute

    2010-02-26

    Terrestrial actinomycetes are noteworthy producers of a multitude of antibiotics, however the marine representatives are much less studied in this regard. In this study, 90 actinomycetes were isolated from 11 different species of marine sponges that had been collected from offshore Ras Mohamed (Egypt) and from Rovinj (Croatia). Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 18 different actinomycete genera representing seven different suborders. Fourteen putatively novel species were identified based on sequence similarity values below 98.2% to other strains in the NCBI database. A putative new genus related to Rubrobacter was isolated on M1 agar that had been amended with sponge extract, thus highlighting the need for innovative cultivation protocols. Testing for anti-infective activities was performed against clinically relevant, Gram-positive (Enterococcus faecalis, Staphylococcus aureus) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria, fungi (Candida albicans) and human parasites (Leishmania major, Trypanosoma brucei). Bioactivities against these pathogens were documented for 10 actinomycete isolates. These results show a high diversity of actinomycetes associated with marine sponges as well as highlight their potential to produce anti-infective agents.

  17. Isolation, Phylogenetic Analysis and Anti-infective Activity Screening of Marine Sponge-Associated Actinomycetes

    PubMed Central

    Abdelmohsen, Usama Ramadan; Pimentel-Elardo, Sheila M.; Hanora, Amro; Radwan, Mona; Abou-El-Ela, Soad H.; Ahmed, Safwat; Hentschel, Ute

    2010-01-01

    Terrestrial actinomycetes are noteworthy producers of a multitude of antibiotics, however the marine representatives are much less studied in this regard. In this study, 90 actinomycetes were isolated from 11 different species of marine sponges that had been collected from offshore Ras Mohamed (Egypt) and from Rovinj (Croatia). Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 18 different actinomycete genera representing seven different suborders. Fourteen putatively novel species were identified based on sequence similarity values below 98.2% to other strains in the NCBI database. A putative new genus related to Rubrobacter was isolated on M1 agar that had been amended with sponge extract, thus highlighting the need for innovative cultivation protocols. Testing for anti-infective activities was performed against clinically relevant, Gram-positive (Enterococcus faecalis, Staphylococcus aureus) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria, fungi (Candida albicans) and human parasites (Leishmania major, Trypanosoma brucei). Bioactivities against these pathogens were documented for 10 actinomycete isolates. These results show a high diversity of actinomycetes associated with marine sponges as well as highlight their potential to produce anti-infective agents. PMID:20411105

  18. Actinomycetes from the South China Sea sponges: isolation, diversity, and potential for aromatic polyketides discovery.

    PubMed

    Sun, Wei; Zhang, Fengli; He, Liming; Karthik, Loganathan; Li, Zhiyong

    2015-01-01

    Marine sponges often harbor dense and diverse microbial communities including actinobacteria. To date no comprehensive investigation has been performed on the culturable diversity of the actinomycetes associated with South China Sea sponges. Structurally novel aromatic polyketides were recently discovered from marine sponge-derived Streptomyces and Saccharopolyspora strains, suggesting that sponge-associated actinomycetes can serve as a new source of aromatic polyketides. In this study, a total of 77 actinomycete strains were isolated from 15 South China Sea sponge species. Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 12 families and 20 genera, among which three rare genera (Marihabitans, Polymorphospora, and Streptomonospora) were isolated from marine sponges for the first time. Subsequently, β-ketoacyl synthase (KSα) gene was used as marker for evaluating the potential of the actinomycete strains to produce aromatic polyketides. As a result, KSα gene was detected in 35 isolates related to seven genera (Kocuria, Micromonospora, Nocardia, Nocardiopsis, Saccharopolyspora, Salinispora, and Streptomyces). Finally, 10 strains were selected for small-scale fermentation, and one angucycline compound was detected from the culture extract of Streptomyces anulatus strain S71. This study advanced our knowledge of the sponge-associated actinomycetes regarding their diversity and potential in producing aromatic polyketides.

  19. Antimicrobial investigation of selected soil actinomycetes isolated from unexplored regions of Kashmir Himalayas, India.

    PubMed

    Shah, Aabid Manzoor; Shakeel-U-Rehman; Hussain, Aehtesham; Mushtaq, Saleem; Rather, Muzafar Ahmad; Shah, Aiyatullah; Ahmad, Zahoor; Khan, Inshad Ali; Bhat, Khursheed Ahmad; Hassan, Qazi Parvaiz

    2017-09-01

    The aim of the present study was to isolate and evaluate the antimicrobial potential of soil actinomycetes of Kashmir Himalayas. The secondary metabolites of actinomycetes are the prominent source of antibiotics. A total of 121 morphologically different actinomycete strains were isolated and screened for antimicrobial activity against various human pathogens. The ethyl acetate extract of fermented broth an actinomycete strain, identified as Streptomyces pratensis exhibited significant antimicrobial activity against Staphylococcus aureus ATCC 29213 with MIC 0.25 μg/ml and Mycobacterium tuberculosis Strain H37Rv with MIC 0.062 μg/ml. The strain S. pratensis IIIM06 was grown on large scale and their broth was extracted with ethyl acetate. The extract was subjected to various chromatography techniques which led to the isolation of four compounds whose structures were established as actinomycin C1, actinomycin C2, actinomycin C3 and actiphenol on the basis of spectral data analysis. Actinomycin C1, C2 and C3 exhibited potent antimicrobial activity against S. aureus as well as M. tuberculosis. The isolated indigenous actinomycetes exhibited good antibacterial activity and the study reveals that IIIM06 is a promising strain and could be of great potential for industrial applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Actinomycetes from the South China Sea sponges: isolation, diversity, and potential for aromatic polyketides discovery

    PubMed Central

    Sun, Wei; Zhang, Fengli; He, Liming; Karthik, Loganathan; Li, Zhiyong

    2015-01-01

    Marine sponges often harbor dense and diverse microbial communities including actinobacteria. To date no comprehensive investigation has been performed on the culturable diversity of the actinomycetes associated with South China Sea sponges. Structurally novel aromatic polyketides were recently discovered from marine sponge-derived Streptomyces and Saccharopolyspora strains, suggesting that sponge-associated actinomycetes can serve as a new source of aromatic polyketides. In this study, a total of 77 actinomycete strains were isolated from 15 South China Sea sponge species. Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 12 families and 20 genera, among which three rare genera (Marihabitans, Polymorphospora, and Streptomonospora) were isolated from marine sponges for the first time. Subsequently, β-ketoacyl synthase (KSα) gene was used as marker for evaluating the potential of the actinomycete strains to produce aromatic polyketides. As a result, KSα gene was detected in 35 isolates related to seven genera (Kocuria, Micromonospora, Nocardia, Nocardiopsis, Saccharopolyspora, Salinispora, and Streptomyces). Finally, 10 strains were selected for small-scale fermentation, and one angucycline compound was detected from the culture extract of Streptomyces anulatus strain S71. This study advanced our knowledge of the sponge-associated actinomycetes regarding their diversity and potential in producing aromatic polyketides. PMID:26483773

  1. Isolation, Screening, and Identification of Novel Isolates of Actinomycetes from India for Antimicrobial Applications

    PubMed Central

    Singh, Vineeta; Haque, Shafiul; Singh, Harshita; Verma, Jyoti; Vibha, Kumari; Singh, Rajbir; Jawed, Arshad; Tripathi, C. K. M.

    2016-01-01

    The search for novel bioactive compounds from the natural environment has rapidly been gaining momentum with the increase in multi-drug resistant (MDR) pathogens. In the present study, the antimicrobial potential of novel actinomycetes has been evaluated by initial screening of six soil samples. Primary and secondary screening was performed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Candida albicans, Candida tropicalis, Trichophyton rubrum, and other MDR bacterial and fungal test strains, thirteen active isolates were selected for further study. Microbial strains were identified on the basis of growth conditions and other biochemical characters. Five most active microbial strains were identified using 16S rRNA sequence homology and designated as Streptomyces xanthophaeus MTCC 11938, Streptomyces variabilis MTCC 12266, Streptomyces xanthochromogenes MTCC 11937, Streptomyces levis EU 124569, and Streptomyces sp. NCIM 5500. Four antibacterial and three antifungal compounds isolated from the above five isolates were purified and partially characterized using UV absorption and IR spectra. Two antibacterial metabolites, belong to chromone and peptide antibiotic, respectively. The antifungal compounds were found to be of non-polyene nature. In conclusion, we study the isolation of novel bacterial strains of actinomycetes for producing novel compounds having antibacterial and antifungal activities from the unexplored agro-ecological niches of India. Also, this study paves the way for further characterization of these isolates of Streptomyces sp. for their optimum utilization for antimicrobial purposes. PMID:27999566

  2. Isolation, Screening, and Identification of Novel Isolates of Actinomycetes from India for Antimicrobial Applications.

    PubMed

    Singh, Vineeta; Haque, Shafiul; Singh, Harshita; Verma, Jyoti; Vibha, Kumari; Singh, Rajbir; Jawed, Arshad; Tripathi, C K M

    2016-01-01

    The search for novel bioactive compounds from the natural environment has rapidly been gaining momentum with the increase in multi-drug resistant (MDR) pathogens. In the present study, the antimicrobial potential of novel actinomycetes has been evaluated by initial screening of six soil samples. Primary and secondary screening was performed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Candida albicans, Candida tropicalis, Trichophyton rubrum, and other MDR bacterial and fungal test strains, thirteen active isolates were selected for further study. Microbial strains were identified on the basis of growth conditions and other biochemical characters. Five most active microbial strains were identified using 16S rRNA sequence homology and designated as Streptomyces xanthophaeus MTCC 11938, Streptomyces variabilis MTCC 12266, Streptomyces xanthochromogenes MTCC 11937, Streptomyces levis EU 124569, and Streptomyces sp. NCIM 5500. Four antibacterial and three antifungal compounds isolated from the above five isolates were purified and partially characterized using UV absorption and IR spectra. Two antibacterial metabolites, belong to chromone and peptide antibiotic, respectively. The antifungal compounds were found to be of non-polyene nature. In conclusion, we study the isolation of novel bacterial strains of actinomycetes for producing novel compounds having antibacterial and antifungal activities from the unexplored agro-ecological niches of India. Also, this study paves the way for further characterization of these isolates of Streptomyces sp. for their optimum utilization for antimicrobial purposes.

  3. Biodiversity, Anti-Trypanosomal Activity Screening, and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges

    PubMed Central

    Cheng, Cheng; MacIntyre, Lynsey; Abdelmohsen, Usama Ramadan; Horn, Hannes; Polymenakou, Paraskevi N.; Edrada-Ebel, RuAngelie; Hentschel, Ute

    2015-01-01

    Marine sponge–associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of < 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50) values <20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348) and Micromonospora (SBT687) were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes. PMID:26407167

  4. Biodiversity, Anti-Trypanosomal Activity Screening, and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges.

    PubMed

    Cheng, Cheng; MacIntyre, Lynsey; Abdelmohsen, Usama Ramadan; Horn, Hannes; Polymenakou, Paraskevi N; Edrada-Ebel, RuAngelie; Hentschel, Ute

    2015-01-01

    Marine sponge-associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of < 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50) values <20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348) and Micromonospora (SBT687) were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes.

  5. [Diversity and bioactivity of actinomycetes isolated from medicinal plant Taxus chinensis and rhizospheric soil].

    PubMed

    Zhang, Panpan; Qin, Sheng; Yuan, Bo; Chen, Yongqiang; Cao, Xiaoying; Jiang, Jihong

    2016-02-04

    In order to provide strains with high activity of anti-phytopathogenic fungi and antitumor activity, we studied the diversity and bioactivity of actinomycetes isolated from medicinal plant Taxus chinensis and rhizospheric soil. Seven selective media were used to isolate actinomycetes. Experiments of anti-phytopathogenic fungi, cytotoxicity activity, and the 16S rRNA gene sequencing of them were carried out to evaluate the diversity and bioactivity. Strains with high activity were identified. A total of 277 actinomycetes were isolated, of which 111 strains were selected and assigned to 6 suborders, 7 families and 8 genera, in which Streptomyces can be divided into 10 groups. The bioactivity testing results indicated that: 30.9% isolates showed activity against at least one of the 12 phytopathogenic fungi; 44.1% strains and 33.3% strains showed cytotoxicity activity with inhibition rate above 40% against stomach cancer cell line SGC-7901 and lung cancer cell line NCI-H460 respectively. Actinomycetes isolated from Taxus chinensis and rhizospheric soil is of high diversity and a good source for the selection of bioactive compounds. Streptomyces KLBMP 2170 is an excellent resource with antifungal and cytotoxicity activity for further studies.

  6. Diversity of actinomycetes isolated from Challenger Deep sediment (10,898 m) from the Mariana Trench.

    PubMed

    Pathom-Aree, Wasu; Stach, James E M; Ward, Alan C; Horikoshi, Koki; Bull, Alan T; Goodfellow, Michael

    2006-06-01

    Thirty-eight actinomycetes were isolated from sediment collected from the Mariana Trench (10,898 m) using marine agar and media selective for actinomycetes, notably raffinose-histidine agar. The isolates were assigned to the class Actinobacteria using primers specific for members of this taxon. The phylogenetic analysis based on 16S rRNA gene sequencing showed that the isolates belonged to the genera Dermacoccus, Kocuria, Micromonospora, Streptomyces, Tsukamurella and Williamsia. All of the isolates were screened for genes encoding nonribosomal peptide and polyketide synthetases. Nonribosomal peptide synthetase sequences were detected in more than half of the isolates and polyketide synthases type I (PKS-I) were identified in five out of 38 strains. The Streptomyces isolates produced several unusual secondary metabolites, including a PKS-I associated product. In initial testing for piezotolerance, the Dermacoccus strain MT1.1 grew at elevated hydrostatic pressures.

  7. Phylogenetic appraisal of antagonistic, slow growing actinomycetes isolated from hypersaline inland solar salterns at Sambhar salt Lake, India

    PubMed Central

    Jose, Polpass Arul; Jebakumar, Solomon Robinson David

    2013-01-01

    Inland solar salterns established in the vicinity of Sambhar Lake are extreme saline environments with high salinity and alkalinity. In view of the fact that microbes inhabiting such extreme saline environments flourish the contemporary bioprospecting, it was aimed to selectively isolate slow growing and rare actinomycetes from the unexplored solar salterns. A total of 14 slow growing actinomycetes were selectively isolated from three composite soil samples of inland solar salterns. Among the isolates, four groups were formed according to similarity of the banding patterns obtained by amplified ribosomal DNA restriction analysis (ARDRA). A subset of representative isolates for each ARDRA group was identified using 16S rDNA sequence based phylogenetic analysis and subsequently the entire isolates were assigned under three different genera; Streptomyces, Pseudonocardia, and Actinoalloteichus. The genus Streptomyces was found to be the dominant among the isolates. Furthermore, rare actinomycete genus Actinoalloteichus was isolated for the first time from solar saltern. Determination of salt-tolerance revealed that certain level of salt-tolerance and moderate halophilism occurs among the actinomycetes isolated from the inland salterns. In addition, all the acinomycetes were screened in two levels to unravel their ability to produce antimicrobial compounds. Significant antimicrobial activity was found among the actinomycetes against a range of bacteria and fungi to worth further characterization of these persuasive actinomycetes and their antimicrobial secondary metabolites. In a nutshell, this study offered a first interesting insight on occurrence of antagonistic rare actinomycetes and streptomycetes in inland solar salterns associated with Sambhar salt Lake. PMID:23847611

  8. Characterization of cellulases of fungal endophytes isolated from Espeletia spp.

    PubMed

    Cabezas, Luisa; Calderon, Carolina; Medina, Luis Miguel; Bahamon, Isabela; Cardenas, Martha; Bernal, Adriana Jimena; Gonzalez, Andrés; Restrepo, Silvia

    2012-12-01

    Endophytes are microorganisms that asymptomatically invade plant tissues. They can stimulate plant growth and/or provide defense against pathogen attacks through the production of secondary metabolites. Most endophyte species are still unknown, and because they may have several applications, the study of their metabolic capabilities is essential. We characterized 100 endophytes isolated from Espeletia spp., a genus unique to the paramo ecosystem, an extreme environment in the Andean mountain range. We evaluated the cellulolytic potential of these endophytes on the saccharification of the oil palm empty fruit bunch (OPEFB). The total cellulolytic activity was measured for each endophyte on filter paper (FPA). In addition, the specific carboxymethyl cellulase (CMCase), exoglucanase, and β-glucosidase activities were determined. We found four fungi positive for cellulases. Of these fungi, Penicillium glabrum had the highest cellulolytic activity after partial purification, with maximal CMCase, exoglucanase and β-glucosidase enzyme activities of 44.5, 48.3, and 0.45 U/ml, respectively. Our data showed that the bioprospection of fungi and the characterization of their enzymes may facilitate the process of biofuel production.

  9. Antibacterial Activities of Actinomycete Isolates Collected from Soils of Rajshahi, Bangladesh

    PubMed Central

    Rahman, Md. Ajijur; Islam, Mohammad Zahidul; Islam, Md. Anwar Ul

    2011-01-01

    This study was performed to isolate actinomycete colonies having antibacterial activity from soil samples collected from different places around Rajshahi, Bangladesh. Thirty actinomycete colonies were isolated in pure culture from five soil samples using Starch-casein-nitrate-agar medium. The isolates were grouped in five color series based on their aerial mycelia color and screened for their antibacterial activity against a range of test bacteria. Sixteen isolates (53.3%) were found to have moderate to high activity against four gram-positive and four gram-negative bacteria. Since many isolates showed inhibitory activity against indicator bacteria, it is suggestive that Bangladeshi soil could be an interesting source to explore for antibacterial secondary metabolites. PMID:21904683

  10. Eliciting antibiotics active against the ESKAPE pathogens in a collection of actinomycetes isolated from mountain soils.

    PubMed

    Zhu, Hua; Swierstra, Jasper; Wu, Changsheng; Girard, Geneviève; Choi, Young Hae; van Wamel, Willem; Sandiford, Stephanie K; van Wezel, Gilles P

    2014-08-01

    The rapid emergence of multidrug-resistant (MDR) bacterial pathogens poses a major threat for human health. In recent years, genome sequencing has unveiled many poorly expressed antibiotic clusters in actinomycetes. Here, we report a well-defined ecological collection of >800 actinomycetes obtained from sites in the Himalaya and Qinling mountains, and we used these in a concept study to see how efficiently antibiotics can be elicited against MDR pathogens isolated recently from the clinic. Using 40 different growth conditions, 96 actinomycetes were identified - predominantly Streptomyces - that produced antibiotics with efficacy against the MDR clinical isolates referred to as ESKAPE pathogens: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and/or Enterobacter cloacae. Antimicrobial activities that fluctuated strongly with growth conditions were correlated with specific compounds, including borrelidin, resistomycin, carbomethoxy-phenazine, and 6,7,8- and 5,6,8-trimethoxy-3-methylisocoumarin, of which the latter was not described previously. Our work provided insights into the potential of actinomycetes as producers of drugs with efficacy against clinical isolates that have emerged recently and also underlined the importance of targeting a specific pathogen.

  11. Isolation and identification of fungal endophytes from grasses on the Oregon coast

    USDA-ARS?s Scientific Manuscript database

    Fungal endophytes have been shown to improve abiotic and biotic stress response in plants. Grasses growing along the Oregon coast are exposed to harsh conditions and may harbor endophytes that enable them to survive and grow under these conditions. Endophytic fungi were isolated from thirty-four gra...

  12. Isolation and identification of bacterial endophytes from grasses along the Oregon coast

    USDA-ARS?s Scientific Manuscript database

    Bacterial endophytes have been shown to improve abiotic and biotic stress responses in plants. Grasses growing along the Oregon coast are exposed to harsh conditions and may harbor endophytes that enable them to survive and grow under these conditions. Bacterial endophytes were isolated from thirty-...

  13. Actinomycetes from solitary wasp mud nest and swallow bird mud nest: isolation and screening for their antibacterial activity.

    PubMed

    Kumar, Vijay; Bharti, Alpana; Gupta, Vivek Kumar; Gusain, Omprakash; Bisht, Gajraj Singh

    2012-03-01

    The aim of this study was to isolate and screen actinomycetes from solitary wasp and swallow bird mud nests for antimicrobial activity. The actinomycetes were isolated from soil of nests of solitary wasp and swallow bird, and identified on the basis of morphological characteristics and molecular biological methods. A total of 109 actinomycetal isolates were obtained from 12 soil samples (6 from each habitat) using two media. The highest number of actinomycetes were recovered on Humic acid vitamin agar media (65.13%, n = 71) as compared to actinomycetes isolation agar media (34.86%, n = 38). The antimicrobial activity of actinomycetes isolates was determined using the agar plug method. Among 109 isolates, 51 isolates (46.78%) showed antibacterial activity by agar plug assay. The morphological and molecular characteristics confirmed that the most of active isolates in both sample belonged to the genus Streptomyces, the other potential genera like Streptosporangium, Actinomadura, Saccharopolyspora, Thermoactinomycetes and Nocardia were also recovered, but in a low frequency. The isolates designated as 8(1), BN-6, MN 2(6), MN 2(7) and MN 9(V) showed most promising activity against various drug resistant bacterial pathogens. It seems that the promising isolates from these unusual/unexplored habitats may prove to be an important step in development of drug for treating multi-drug resistant bacterial pathogens.

  14. Antimicrobial activities of actinomycetes isolated from unexplored regions of Sundarbans mangrove ecosystem.

    PubMed

    Sengupta, Sohan; Pramanik, Arnab; Ghosh, Abhrajyoti; Bhattacharyya, Maitree

    2015-08-21

    New broad spectrum antimicrobial agents are urgently needed to combat frequently emerging multi drug resistant pathogens. Actinomycetes, the most talented group of microorganisms isolated from unexplored regions of the world may be the ultimate solution to this problem. Thus the aim of this study was to isolate several bioactive actinomycetes strains capable of producing antimicrobial secondary metabolite from Sundarbans, the only mangrove tiger land of the world. Fifty four actinomycetes were isolated and analyzed for antimicrobial activity against fifteen test organisms including three phytopathogens. Nine morphologically distinct and biologically active isolates were subjected to polyphasic identification study.16 s rDNA sequencing indicated eight isolates to reveal maximum similarity to the genus streptomyces, whereas one isolate presented only 93.57% similarity with Streptomyces albogriseolus NRRL B-1305(T). Seventy-one carbon sources and twenty-three chemical sources utilization assay revealed their metabolic relatedness. Among these nine isolates three specific strains were found to have notably higher degree of antimicrobial potential effective in a broader range including phyto-pathogenic fungus. Finally the strain SMS_SU21, which showed antimicrobial activity with MIC value of 0.05 mg ml(-1) and antioxidant activity with IC50 value of 0.242 ± 0.33 mg ml(-1) was detected to be the most potential one. True prospective of this strain was evaluated utilizing GC-MS and the bioactive compound responsible for antimicrobial activity was purified. Rare bioactive actinomycetes were isolated from unexplored heritage site. Antimicrobial compound has also been identified and purified which is active against a broad range of pathogens.

  15. Application of a method incorporating differential centrifugation for selective isolation of motile actinomycetes in soil and plant litter.

    PubMed

    Hayakawa, M; Otoguro, M; Takeuchi, T; Yamazaki, T; Iimura, Y

    2000-08-01

    The present paper describes a simple enrichment technique which enables rapid and selective isolation of diverse zoosporic actinomycete genera directly from soil and plant litter. This technique, designated the rehydration and centrifugation (RC) method, consists of immersing the air-dried source material in 10 mM phosphate buffer containing 10% soil extract, letting the preparation stand at 30 degrees C for 90 min, followed by centrifugation of the fluid at 1,500 x g for 20 min. Portions of the supernatant containing actinomycete zoospores are plated on the humic acid-vitamin agar which is supplemented with nalidixic acid and trimethoprim as the selective inhibitors for Gram-negative bacteria and bacilli. The phosphate buffer-soil extract solution significantly promoted liberation of motile zoospores from the source material. The centrifugation stage greatly eliminated streptomycetes and other non-motile actinomycetes from the liquid phase, thereby facilitating selective growth of rare, motile actinomycetes on the isolation plates subsequent to inoculation. Ten different soil and leaf-litter samples, taken from fields, forests, and stream banks, were examined. The RC method consistently achieved preferential isolation of motile actinomycetes in all samples, which accounted for 37-86% of the total microbial population recovered. The most frequently isolated motile actinomycetes were Actinoplanes and Dactylosporangium. Strains of Actinokineospora, Catenuloplanes and Kineosporia were also recovered, depending on the nature of the samples examined. Other motile actinomycetes that were occasionally isolated in small numbers included Actinosynnema, Geodermatophilus and Sporichthya.

  16. Antimicrobial compounds from endophytic Streptomyces sp. BCC72023 isolated from rice (Oryza sativa L.).

    PubMed

    Supong, Khomsan; Thawai, Chitti; Choowong, Wilunda; Kittiwongwattana, Chokchai; Thanaboripat, Dusanee; Laosinwattana, Chamroon; Koohakan, Prommart; Parinthawong, Nonglak; Pittayakhajonwut, Pattama

    2016-05-01

    An endophytic actinomycete strain BCC72023 was isolated from rice (Oryza sativa L.) and identified as the genus Streptomyces, based on phenotypic, chemotaxonomic and 16S rRNA gene sequence analyses. The strain showed 99.80% similarity compared with Streptomyces samsunensis M1463(T). Chemical investigation led to the isolation of three macrolides, efomycins M (1), G (2) and oxohygrolidin (3), along with two polyethers, abierixin (4) and 29-O-methylabierixin (5). To our knowledge, this is the first report of efomycin M being isolated from a natural source. The compounds were identified using spectroscopic techniques and comparison with previously published data. All compounds exhibited antimalarial activity against the Plasmodium falciparum, K-1 strain, a multidrug-resistant strain, with IC50 values in a range of 1.40-5.23 μg/ml. In addition, these compounds were evaluated for biological activity against Mycobacterium tuberculosis, Bacillus cereus, Colletotrichum gloeosporioides and Colletotrichum capsici, as well as cytotoxicity against both cancerous (MCF-7, KB, NCI-H187) and non-cancerous (Vero) cells.

  17. Bioprospecting of Novel and Bioactive Compounds from Marine Actinomycetes Isolated from South China Sea Sediments.

    PubMed

    Yang, Na; Song, Fuhang

    2017-09-16

    Marine actinomycetes are less investigated compared to terrestrial strains as potential sources of natural products. To date, few investigations have been performed on culturable actinomycetes associated with South China Sea sediments. In the present study, twenty-eight actinomycetes were recovered from South China Sea sediments after dereplication by traditional culture-dependent method. The 16S rRNA gene sequences analyses revealed that these strains related to five families and seven genera. Twelve representative strains possessed at least one of the biosynthetic genes coding for polyketide synthase I, II, and nonribosomal peptide synthetase. Four strains had anti-Mycobacterium phlei activities and five strains had activities against methicillin-resistant Staphylococcus aureus. 10 L-scale fermentation of strains Salinispora sp. NHF45, Nocardiopsis sp. NHF48, and Streptomyces sp. NHF86 were carried out for novel and bioactive compounds discovery. Finally, we obtained a novel α-pyrone compound from marine Nocardiopsis sp. NHF48, an analogue of paulomenol from marine Streptomyces sp. NHF86 and a new source of rifamycin B, produced by Salinispora sp. NHF45. The present study concluded that marine actinomycetes, which we isolated from South China Sea sediments, will be a suitable source for the development of novel and bioactive compounds.

  18. Isolation and screening of antibiotic producing actinomycetes from soils in Gondar town, North West Ethiopia

    PubMed Central

    Bizuye, Abebe; Moges, Feleke; Andualem, Berhanu

    2013-01-01

    Objective To isolate and screen antibiotic producing actinomycetes from potential soil samples of Gondar town, Ethiopia. Methods Fifteen soil samples were collected, serially diluted and spread on starch casein and oat meal agar supplemented with amoxicillin and cyclohexamide for inhibition of bacteria and fungi, respectively. Cross streak method was used to check antagonistic activity of isolated actinomycetes against test organisms. Solid state fermentation and crude extraction were used for the production of antibiotics from isolates. Agar well diffusion was used for antimicrobial activity of crude extracts against test organisms. Results Three isolates (Ab18, Ab28 and Ab43) have been shown high antagonistic activity during primary screening. Inhibition zones obtained from crude extracts showed significance differences when compared with standard antibiotics tested against test organisms (P<0.05). Inhibition zone of crude extracts from isolate Ab18 against Klebsiella pneumonia ATCC7000603 and Escherichia coli ATCC25922 were (14±1) mm and (35±1) mm, respectively which were strong active when compared to amoxicillin (0 mm) and tetracycline [(13±1) mm for Klebsiella pneumonia ATCC7000603 and (33±1) mm for Escherichia coli ATCC25922]. Crude extracts from isolate Ab18 showed (20±1) mm and (15±1) mm inhibition zones against methicillin resistant Staphylococcus aureus strains 2 (MRSA2) and MRSA4, respectively. Crude extract from isolate Ab43 has shown inhibition zones of (16±1) mm and (17±1) mm against MRSA2 and MRSA4, respectively. Combination of Ab18 and Ab43 has shown high antimicrobial activity (18±1) mm against MRSA2 and MRSA4. Conclusions There was not any scientific report on soil actinomycetes producing antibiotic in the study areas. Therefore, isolation and screening of actinomycetes from such areas in optimum condition may contribute the discovery of new antibiotics. Potent antibiotics from these actinomycetes could contribute a lot to fight against

  19. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    PubMed Central

    Bernal, Milagro García; Campa-Córdova, Ángel Isidro; Saucedo, Pedro Enrique; González, Marlen Casanova; Marrero, Ricardo Medina; Mazón-Suástegui, José Manuel

    2015-01-01

    Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4) among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world. PMID:27047067

  20. Chromium(VI) resistance and removal by actinomycete strains isolated from sediments.

    PubMed

    Polti, Marta A; Amoroso, María J; Abate, Carlos M

    2007-03-01

    Forty-one isolated actinomycetes were used to study qualitative and semi-quantitative screening of chromium(VI) resistance. Chromate-removing activity was estimated using the Cr(VI) specific colorimetric reagent 1,5-diphenylcarbazide. Twenty percent of the isolates from El Cadillal (EC) and 14% of isolates from a copper filter plant (CFP) were able to grow at 13 mM of Cr(VI). All isolates from sugar cane (SCP) could grow up to Cr(VI) concentration of 17 mM. EC, CFP and SCP strains were able to remove 24%, 30% and more than 40% of Cr(VI), respectively. The highest and lowest Cr(VI) specific removal values were 75.5 mg g(-1) cell by M3 (CFP), and 1.5 mg g(-1) cell by C35 (EC) strains. Eleven Cr(VI) resistant strains were characterized and identified as species of the genera Streptomyces (10) and Amycolatopsis (1). Differences on actinomycete community composition between contaminated and non-contaminated soil were found. This study showed the potential capacity of actinomycetes as tools for Cr(VI) bioremediation.

  1. Non-Frankia Actinomycetes Isolated from Surface-Sterilized Roots of Casuarina equisetifolia Fix Nitrogen†

    PubMed Central

    Valdés, María; Pérez, Néstor-Octavio; Estrada-de los Santos, Paulina; Caballero-Mellado, Jesús; Peña-Cabriales, Juan José; Normand, Philippe; Hirsch, Ann M.

    2005-01-01

    Based on partial 16S sequences, we previously described a novel group of nonsymbiotic, acetylene reduction activity-positive actinomycetes which were isolated from surface-sterilized roots of Casuarina equisetifolia growing in Mexico. An amplified rRNA restriction analysis confirmed that these actinomycetes are distinct from Frankia, a finding substantiated by a 16S rRNA gene phylogenetic analysis of two of the Mexican isolates. Further support for these actinomycetes being separate from Frankia comes from the very low DNA-DNA homology that was found. Nevertheless, the Mexican isolates may be diazotrophs based not only on their ability to grow in N-free medium and reduce acetylene to ethylene but also on the results from 15N isotope dilution analysis and the finding that a nifH gene was PCR amplified. A comparison of the nifH sequences from the various isolates showed that they are closely related to nifH from Frankia; the similarity was 84 to 98% depending on the host specificity group. An analysis of complete 16S rRNA gene sequences demonstrated that the two strains analyzed in detail are most closely related to actinobacteria in the Thermomonosporaceae and the Micromonosporaceae. PMID:15640222

  2. Isolation and characterization of novel marine-derived actinomycete taxa rich in bioactive metabolites.

    PubMed

    Magarvey, Nathan A; Keller, Jessica M; Bernan, Valerie; Dworkin, Martin; Sherman, David H

    2004-12-01

    A unique selective enrichment procedure has resulted in the isolation and identification of two new genera of marine-derived actinobacteria. Approximately 90% of the microorganisms cultured by using the presented method were from the prospective new genera, a result indicative of its high selectivity. In this study, 102 actinomycetes were isolated from subtidal marine sediments collected from the Bismarck Sea and the Solomon Sea off the coast of Papua New Guinea. A combination of physiological parameters, chemotaxonomic characteristics, distinguishing 16S rRNA gene sequences, and phylogenetic analysis based on 16S rRNA genes provided strong evidence for the two new genera (represented by strains of the PNG1 clade and strain UMM518) within the family Micromonosporaceae. Biological activity testing of fermentation products from the new marine-derived actinomycetes revealed that several had activities against multidrug-resistant gram-positive pathogens, malignant cells, and vaccinia virus replication.

  3. Isolation and Characterization of Novel Marine-Derived Actinomycete Taxa Rich in Bioactive Metabolites†

    PubMed Central

    Magarvey, Nathan A.; Keller, Jessica M.; Bernan, Valerie; Dworkin, Martin; Sherman, David H.

    2004-01-01

    A unique selective enrichment procedure has resulted in the isolation and identification of two new genera of marine-derived actinobacteria. Approximately 90% of the microorganisms cultured by using the presented method were from the prospective new genera, a result indicative of its high selectivity. In this study, 102 actinomycetes were isolated from subtidal marine sediments collected from the Bismarck Sea and the Solomon Sea off the coast of Papua New Guinea. A combination of physiological parameters, chemotaxonomic characteristics, distinguishing 16S rRNA gene sequences, and phylogenetic analysis based on 16S rRNA genes provided strong evidence for the two new genera (represented by strains of the PNG1 clade and strain UMM518) within the family Micromonosporaceae. Biological activity testing of fermentation products from the new marine-derived actinomycetes revealed that several had activities against multidrug-resistant gram-positive pathogens, malignant cells, and vaccinia virus replication. PMID:15574955

  4. Isolation of bacterial endophytes from germinated maize kernels.

    PubMed

    Rijavec, Tomaz; Lapanje, Ales; Dermastia, Marina; Rupnik, Maja

    2007-06-01

    The germination of surface-sterilized maize kernels under aseptic conditions proved to be a suitable method for isolation of kernel-associated bacterial endophytes. Bacterial strains identified by partial 16S rRNA gene sequencing as Pantoea sp., Microbacterium sp., Frigoribacterium sp., Bacillus sp., Paenibacillus sp., and Sphingomonas sp. were isolated from kernels of 4 different maize cultivars. Genus Pantoea was associated with a specific maize cultivar. The kernels of this cultivar were often overgrown with the fungus Lecanicillium aphanocladii; however, those exhibiting Pantoea growth were never colonized with it. Furthermore, the isolated bacterium strain inhibited fungal growth in vitro.

  5. Isolation and Characterisation of Endophytic Nitrogen Fixing Bacteria in Sugarcane.

    PubMed

    Muangthong, Ampiga; Youpensuk, Somchit; Rerkasem, Benjavan

    2015-04-01

    Endophytic nitrogen fixing bacteria were isolated from the leaves, stems and roots of industrial variety (cv. U-Thong 3; UT3), wild and chewing sugarcane plants grown for 6 weeks in nitrogen (N)-free sand. Eighty nine isolates of endophytic bacteria were obtained on N-free agar. An acetylene reduction assay (ARA) detected nitrogenase activity in all 89 isolates. Three isolates from the chewing (C2HL2, C7HL1 and C34MR1) sugarcane and one isolate from the industrial sugarcane (UT3R1) varieties were characterised, and their responses to different yeast extract concentrations were investigated. Three different responses in nitrogenase activity were observed. Isolates C2HL2 and C7HL1 exhibited major increases with the addition of 0.005% yeast extract, C34MR1 exhibited no response, and UT3R1 exhibited a significant decrease in nitrogenase activity with 0.005% yeast extract. In all the isolates, nitrogenase activity decreased with further increase of the yeast extract to 0.05%. The highest nitrogenase activity was observed in isolates C2HL2 and C7HL1, which had 16S rRNA gene sequences that were closely related to Novosphingobium sediminicola and Ochrobactrum intermedium, respectively.

  6. Biological Characteristics and Antimicrobial Activity of Endophytic Streptomyces sp. TQR12-4 Isolated from Elite Citrus nobilis Cultivar Ham Yen of Vietnam

    PubMed Central

    Mai-Linh, Nguyen Vu; Hong-Lien, Nguyen Thi; Van Hieu, Nguyen

    2016-01-01

    Ham Yen orange (Citrus nobilis Lour) is the highly valuable commercial fruit of Vietnam. With the blooming of fruit production and farming area, this specialty crop is facing threats from several serious diseases; therefore the search for new effective biocontrollers is required to prevent the existing excessive use of fertilizers and plant protection chemicals. Endophytic actinomycetes are of great scientific interest due to their high potential of application in agriculture and pharmaceutical research. In this work, endophytic actinomycetes were isolated from a native orange species of Northeast mountainous province Tuyen Quang. Among 49 isolates obtained, the isolate TQR12-4 strongly inhibited test pathogens Colletotrichum truncatum, Geotrichum candidum, Fusarium oxysporum, and F. udum. This isolate gave comparatively high biomass yields on different substrates, for example, carboxy methyl cellulose, starch, protein, and chitin, within a wide range of temperature from 15 to 45°C and pH from 4 to 10. Sequence analysis of 16S rDNA gene showed that TQR12-4 shared 99% similarity to Streptomyces prasinopilosus; however, it slightly differed from the latter in spore morphology and hence was named as Streptomyces sp. TQR12-4. A thermostable antifungal substance of nonpeptide nature produced by Streptomyces sp. TQR12-4 had MIC against Fusarium udum of 100 μg/mL and 400 μg/mL respective to extract fractions X4 and X5. PMID:27795709

  7. Novel actinomycete and a cyanide-degrading pseudomonad isolated from industrial sludge

    SciTech Connect

    White, J.M.

    1987-01-01

    A novel actinomycete was the predominant filamentous microorganism in bulking activated sludge in a bench-scale reactor treating coke plant wastewater. The bacterium was isolated and identified as an actinomycete that is biochemically and morphologically similar to Amycolatopsis orientalis; however, a lack of DNA homology excludes true relatedness. At present, the isolate (NRRL B 16216) cannot be assigned to the recognized taxa of actinomycetes. Cyanide-degrading microorganisms were selected in chemostats maintained at a low dilution rate for several weeks. Cyanide alone or cyanide plus phenol were fully degraded when equilibrium was achieved, and increasing concentrations of cyanide were degraded until inhibition of cell division resulted in cell washout. An isolated non-fluorescent pseudomonad could be adapted to degrade high concentrations of cyanide and to utilize cyanide-nitrogen when phenol or lactate was the carbon source. Although one-carbon compounds such as methanol and methylamine were growth substrates, cyanide was not utilized as a carbon source. In the absence of cyanide, adaptation was gradually lost. Oxygen consumption of adapted cells was stimulated in the presence of cyanide whereas that of unadapted cells was depressed. Cyanide was degraded by growing or resting cells and by cell-free extracts. Cyanide degrading activity of cell-free extracts, lost upon dialysis, was fully restored with NAD(P)H.

  8. Isolation and characterization of potential antibiotic producing actinomycetes from water and sediments of Lake Tana, Ethiopia.

    PubMed

    Gebreyohannes, Gebreselema; Moges, Feleke; Sahile, Samuel; Raja, Nagappan

    2013-06-01

    To isolate, evaluate and characterize potential antibiotic producing actinomycetes from water and sediments of Lake Tana, Ethiopia. A total of 31 strains of actinomycetes were isolated and tested against Gram positive and Gram negative bacterial strains by primary screening. In the primary screening, 11 promising isolates were identified and subjected to solid state and submerged state fermentation methods to produce crude extracts. The fermented biomass was extracted by organic solvent extraction method and tested against bacterial strains by disc and agar well diffusion methods. The isolates were characterized by using morphological, physiological and biochemical methods. The result obtained from agar well diffusion method was better than disc diffusion method. The crude extract showed higher inhibition zone against Gram positive bacteria than Gram negative bacteria. One-way analysis of variance confirmed most of the crude extracts were statistically significant at 95% confidence interval. The minimum inhibitory concentration and minimum bactericidal concentration of crude extracts were 1.65 mg/mL and 3.30 mg/mL against Staphylococcus aureus, and 1.84 mg/mL and 3.80 mg/mL against Escherichia coli respectively. The growth of aerial and substrate mycelium varied in different culture media used. Most of the isolates were able to hydrolysis starch and urea; able to survive at 5% concentration of sodium chloride; optimum temperature for their growth was 30 °C. The results of the present study revealed that freshwater actinomycetes of Lake Tana appear to have immense potential as a source of antibacterial compounds.

  9. Isolation and characterization of Cr(VI)-reducing actinomycetes from estuarine sediments.

    PubMed

    Terahara, Takeshi; Xu, Xudan; Kobayashi, Takeshi; Imada, Chiaki

    2015-04-01

    Bioremediation technologies have strong potential use in the less costly and more environmentally friendly removal of highly toxic hexavalent-chromium (Cr(VI)) compared with physicochemical technologies. Several Cr(VI)-reducing bacteria have been isolated; however, there are few studies on Cr(VI)-resistant and Cr(VI)-reducing actinomycetes. In this study, Cr(VI)-reducing actinomycetes were screened from estuarine, marine, and terrestrial samples on the basis of Cr(VI)-resistant and Cr(VI)-reducing ability. Of the 80 Streptomyces-like strains isolated, 20 strains were found to be resistant to 50 mg/l of Cr(VI). In addition, two strains isolated from the estuarine sediment of Tokyo Bay were found to be resistant to a concentration of 150 mg/l of Cr(VI). Furthermore, one Cr(VI)-reducing strain was found to remove 60 mg/l of Cr(VI) within 1 week and was identified as Streptomyces thermocarboxydus based on 16S rRNA gene analysis. The comparative evaluation with the type strain S. thermocarboxydus NBRC 16323 showed that our isolated strain had higher ability to grow at 27 °C and reduce Cr(VI) at a NaCl concentration of 6.0 % at 27 °C compared with the type strain NBRC 16323. These results indicate that our isolated strain have a potential ability to remove Cr(VI) from contaminated, highly saline sources without heating.

  10. Biodegradation of anthracene by a novel actinomycete, Microbacterium sp. isolated from tropical hydrocarbon-contaminated soil.

    PubMed

    Salam, Lateef B; Obayori, Oluwafemi S; Olatoye, Nojeem O

    2014-01-01

    A novel anthracene-degrading Gram-positive actinomycete, Microbacterium sp. strain SL10 was isolated from a hydrocarbon-contaminated soil at a mechanical engineering workshop in Lagos, Nigeria. The polluted soil had an unusually high total hydrocarbon content of 157 g/kg and presence of various heavy metals. The isolate tolerated salt concentration of more than 4%. It resisted cefotaxime, streptomycin and ciprofloxacin, but susceptible to meropenem, linezolid and vancomycin. The isolate exhibited growth rate and doubling time of 0.82 days(-1) and 0.84 days, respectively on anthracene. It degraded 57.5 and 90.12% of anthracene within 12 and 21 days, respectively while the rate of anthracene utilization by the isolate was 4.79 mg l(-1) d(-1). To the best of our knowledge, this is the first report of isolation and characterization of anthracene-degrading Microbacterium sp.

  11. Actinomadura xylanilytica sp. nov., an actinomycete isolated from soil.

    PubMed

    Zucchi, Tiago Domingues; Kim, Byung-Yong; Bonda, Avinash Naga Venkata; Goodfellow, Michael

    2013-02-01

    The taxonomic position of a soil isolate, strain BK147(T), was established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Actinomadura. It formed a distinct phyletic line in the phylogenetic tree based on 16S rRNA gene sequences of members of the genus Actinomadura and was most closely, albeit loosely, related to Actinomadura bangladeshensis DSM 45347(T), Actinomadura meyerae DSM 44715(T) and Actinomadura napierensis NRRL B-24319(T) but was readily distinguished from these strains using a range of phenotypic properties. Based on the combined genotypic and phenotypic data it is proposed that isolate BK147(T) ( = KACC 20919(T) = NCIMB 14771(T) = NRRL B-24852(T)) be classified as the type strain of a novel species of the genus Actinomadura, for which the name Actinomadura xylanilytica sp. nov. is proposed.

  12. Rhodococcus kyotonensis sp. nov., a novel actinomycete isolated from soil.

    PubMed

    Li, Bing; Furihata, Keiko; Ding, Lin-Xian; Yokota, Akira

    2007-09-01

    A polyphasic study was undertaken to establish the taxonomic position of an isolate, strain DS472(T), from soil in Kyoto, Japan. Phylogenetic analysis, based on the 16S rRNA gene sequences, revealed that this strain constitutes a new subline within the genus Rhodococcus, with Rhodococcus yunnanensis YIM 70056(T) and Rhodococcus fascians DSM 20669(T) as its nearest phylogenetic neighbours (98.2 and 97.8 % sequence similarity, respectively). DNA-DNA hybridization experiments revealed 36 and 29 % relatedness between the isolate and its phylogenetic relatives, R. yunnanensis and R. fascians, respectively. Chemotaxonomic characteristics, including the major quinone MK-8(H(2)), predominant fatty acids C(16 : 0), C(18 : 1)omega9c and 10-methyl C(18 : 0), the presence of cell-wall chemotype IV and mycolic acids, were consistent with the properties of members of the genus Rhodococcus. The DNA G+C content was 64.5 mol%. On the basis of both phenotypic and genotypic evidence, strain DS472(T) represents a novel species of the genus Rhodococcus, for which the name Rhodococcus kyotonensis sp. nov. is proposed. The type strain is strain DS472(T) (=IAM 15415(T)=CCTCC AB206088(T)).

  13. [Study of the lytic activity of actinomycetes isolated from various soils in Georgia].

    PubMed

    Kudukhashvili, P G; Gurielidze, M A; Pataraia, D T

    2001-01-01

    The lytic activities of 310 cultures from the Collection of Actinomycetes of the Institute of Biochemistry and Biotechnology, National Academy of Sciences of Georgia, were studied; 18% of these strains appeared capable of lysing the yeast cell wall. The active producer of the enzyme was selected. This culture was isolated from chestnut soil in Gardabani raion (Central Georgia). Its cultural-morphological, biochemical, and antagonistic properties allowed the culture to be ascribed to the species Geodermatophilus obseurus Luedemann, 1968. The maximal lytic activity under deep cultivation conditions, exceeding twofold the activity of Actinomyces griseinus, was observed during the logarithmic growth phase.

  14. Streptomyces luteus sp. nov., an actinomycete isolated from soil.

    PubMed

    Luo, Xiao-Xia; Kai, Liang; Wang, Yang; Wan, Chuan-Xing; Zhang, Li-Li

    2017-04-01

    A Streptomyces-like strain, designated TRM 45540T, was isolated from soil of the Loulan area (89° 22' 22″ E 40° 29' 55″ N), Xinjiang Province, north-west China, and was characterized taxonomically by using a polyphasic study. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain TRM 45540T shared 99.87 % similarity with Streptomyces mutabilis NBRC 12800T (GenBank accession number AB184156). The strain was aerobic, Gram-stain-positive, with an optimum NaCl concentration for growth of 5 % (w/v). The isolate formed white aerial mycelium that was long filamentous with few branches; the substrate mycelium possessed long, smooth-surfaced spore chains bearing smooth spores and produced a yellow diffusible pigment. The strain contained iso-C16 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and C16 : 0 as major cellular fatty acids. The predominant menaquinones of the strain were MK-9(H6), MK-9(H4) and MK-9(H10). The whole-cell sugar pattern contained glucose and ribose. The polar lipid pattern of the strain consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphotidylinositol, phosphatidylglycerol and phosphatidylinositolmannosides. Genotypic and phenotypic data confirmed that strain TRM 45540T represents a novel species, clearly different from related species of the genus Streptomyces, and for which the name Streptomyces luteus (type strain TRM 45540T=CCTCC AA 2014003T=NRRL B-59117T) is proposed.

  15. Presence, molecular characteristics and geosmin producing ability of actinomycetes isolated from South Korean terrestrial and aquatic environments.

    PubMed

    Lee, Gyu-Cheol; Kim, Yun S; Kim, Min-Jeong; Oh, Sung-Ae; Choi, Ilhwan; Choi, Jaewon; Park, Jong-Geun; Chong, Chom-Kyu; Kim, Yong-Yeon; Lee, Kyeunghee; Lee, Chan Hee

    2011-01-01

    The unpleasant odor of drinking water is one of the major problems in many water utilities in the world. Actinomycetes have long been associated with odorous compounds. Considering the paucity of research on Actinomycetes producing odorous compounds in South Korea, presence of Actinomycetes, their molecular characteristics and ability to produce odorous compounds were investigated in this study. Findings confirmed the presence of Actinomycetes in surface soil, sediment, and water samples from four sites: two artificial lakes [Paldang and Cheongpyeong (CP)], and two streams [Gyeongan (GA) and Yangpyeong]. Surface soil and sediment from CP area had the greatest concentration of Actinomycetes (8.2 x 10(7) and 6.8 x 10(6) colony forming units (CFUs)/gram, dry weight, respectively). When water samples are considered, samples from GA had the highest concentration (1.9 x 10(2) CFU/mL). 16S rRNA sequencing and molecular phylogenetic analysis showed that Streptomyces was the dominant genus (64.1%). In addition, the isolated Actinomycetes synthesized 5.4 ng/L geosmin as demonstrated by thermal desorption unit-gas chromatograph/mass spectrometry analysis.

  16. Siderophore production by actinomycetes isolates from two soil sites in Western Australia.

    PubMed

    Lee, Joanna; Postmaster, Armin; Soon, Hooi Peng; Keast, David; Carson, Kerry C

    2012-04-01

    The actinomycetes are metabolically flexible soil micro-organisms capable of producing a range of compounds of interest, including siderophores. Siderophore production by actinomycetes sampled from two distinct and separate geographical sites in Western Australia were investigated and found to be generally similar in the total percentage of siderophore producers found. The only notable difference was the proportion of isolates producing catechol siderophores with only 3% found in site 1 (from the north-west of Western Australia and reportedly containing 40% magnetite) and 17% in site 2 (a commercial stone fruit orchard in the hills east of Perth with a soil base ranging from sandy loam to laterite). Further detailed characterization of isolates of interest identified a Streptomyces that produced extracellularly excreted enterobactin, the characteristic Enterobacteriaceae siderophore, and also revealed some of the conditions required for enterobactin production. Carriage of the entF gene, which codes for the synthetase responsible for the final assembly of the tri-cyclic structure of enterobactin, was confirmed by PCR in this isolate. Another separate Streptomyces produced a compound that matched the UV/VIS spectra of heterobactin, a siderophore previously only described in Rhodococcus and Nocardia.

  17. Homoharringtonine production by endophytic fungus isolated from Cephalotaxus hainanensis Li.

    PubMed

    Hu, Xiaoping; Li, Wu; Yuan, Mu; Li, Congfa; Liu, Sixin; Jiang, Chunjie; Wu, Yanchun; Cai, Kun; Liu, Yan

    2016-07-01

    Homoharringtonine (HHT), a natural plant alkaloid derived from Cephalotaxus, has demonstrated to have a broad antitumor activity and efficacy in treating human chronic myeloid leukemia. An alternative source is required to substitute for the slow-growing and scarce Cephalotaxus to meet the increasing demand of the drug market. The objective of this study was to screen HHT-producing endophytic fungi from Cephalotaxus hainanensis Li. By screening 213 fungal isolates obtained from the bark parts of Cephalotaxus hainanensis Li, one isolate was found to be capable of biosynthesizing HHT. The fungus was identified as Alternaria tenuissima by morphological characteristics and internal transcribed spacer (ITS) sequence analysis and was named as CH1307. HHT obtained from CH1307 was analyzed through the HPLC and LC-MS/MS and NMR spectroscopy. The extract of the fermentation broth of CH1307 showed antiproliferative activities against K562 (chronic myelocytic leukemia), NB4 (acute promyelocytic leukemia), and HL-60 (promyelocytic leukemia) human cancer cell lines with IC50 values of 67.25 ± 4.26, 65.02 ± 4.75, and 99.23 ± 4.26 μg/mL, respectively. The findings suggest that HHT-producing endophytic fungus, Alternaria tenuissima CH1307 might provide a promising source for the research and application of HHT.

  18. Isolation and characterization of endophytic bacteria isolated from the leaves of the common bean (Phaseolus vulgaris)

    PubMed Central

    de Oliveira Costa, Leonardo Emanuel; de Queiroz, Marisa Vieira; Borges, Arnaldo Chaer; de Moraes, Celia Alencar; de Araújo, Elza Fernandes

    2012-01-01

    The common bean is one of the most important legumes in the human diet, but little is known about the endophytic bacteria associated with the leaves of this plant. The objective of this study was to characterize the culturable endophytic bacteria of common bean (Phaseolus vulgaris) leaves from three different cultivars (Vermelhinho, Talismã, and Ouro Negro) grown under the same field conditions. The density of endophytic populations varied from 4.5 x 102 to 2.8 x 103 CFU g-1 of fresh weight. Of the 158 total isolates, 36.7% belonged to the Proteobacteria, 32.9% to Firmicutes, 29.7% to Actinobacteria, and 0.6% to Bacteroidetes. The three P. vulgaris cultivars showed class distribution differences among Actinobacteria, Alphaproteobacteria and Bacilli. Based on 16S rDNA sequences, 23 different genera were isolated comprising bacteria commonly associated with soil and plants. The genera Bacillus, Delftia, Methylobacterium, Microbacterium, Paenibacillus, Staphylococcus and Stenotrophomonas were isolated from all three cultivars. To access and compare the community structure, diversity indices were calculated. The isolates from the Talismã cultivar were less diverse than the isolates derived from the other two cultivars. The results of this work indicate that the cultivar of the plant may contribute to the structure of the endophytic community associated with the common bean. This is the first report of endophytic bacteria from the leaves of P. vulgaris cultivars. Future studies will determine the potential application of these isolates in biological control, growth promotion and enzyme production for biotechnology. PMID:24031988

  19. In vitro α-glucosidase inhibition and antioxidative potential of an endophyte species (Streptomyces sp. loyola UGC) isolated from Datura stramonium L.

    PubMed

    Nimal Christhudas, I V S; Praveen Kumar, P; Agastian, P

    2013-07-01

    Endophytic actinomycetes isolated from Datura stramonium L. was evaluated for its effects against in vitro α-glucosidase inhibition, antioxidant, and free radical scavenging activities. Based on microbial cultural characteristic and 16S rRNA sequencing, it was identified as Streptomyces sp. loyola UGC. The methanolic extract of endophytic actinomycetes (MeEA) shows remarkable inhibition of α-glucosidase (IC50 730.21 ± 1.33 μg/ml), scavenging activity on 2,2-diphenyl-picrylhydrazyl (DPPH) (IC50 435.31 ± 1.79 μg/ml), hydroxyl radical (IC50 350.21 ± 1.02 μg/ml), nitric oxide scavenging (IC50 800.12 ± 1.05 μg/ml), superoxide anion radical (IC50 220.31 ± 1.47 μg/ml), as well as a high and dose-dependent reducing power. The MeEA also showed a strong suppressive effect on rat liver lipid peroxidation. Antioxidants of β-carotene linoleate model system revels significantly lower than BHA. The total phenolic content of the extract was 176 mg of catechol equivalents/gram extract. Perusal of this study indicates MeEA can be used as natural resource of α-glucosidase inhibitor and antioxidants.

  20. Amycolatopsis saalfeldensis sp. nov., a novel actinomycete isolated from a medieval alum slate mine.

    PubMed

    Carlsohn, Marc René; Groth, Ingrid; Tan, Geok Yuan Annie; Schütze, Barbara; Saluz, Hans-Peter; Munder, Thomas; Yang, Jun; Wink, Joachim; Goodfellow, Michael

    2007-07-01

    Three actinomycetes isolated from the surfaces of rocks in a medieval slate mine were examined in a polyphasic taxonomic study. Chemotaxonomic and morphological characteristics of the isolates were typical of strains of the genus Amycolatopsis. The isolates had identical 16S rRNA gene sequences and formed a distinct phyletic line towards the periphery of the Amycolatopsis mediterranei clade, being most closely related to Amycolatopsis rifamycinica. The organisms shared a wide range of genotypic and phenotypic markers that distinguished them from their closest phylogenetic neighbours. On the basis of these results, a novel species, Amycolatopsis saalfeldensis sp. nov., is proposed. The type strain is HKI 0457(T) (=DSM 44993(T)=NRRL B-24474(T)).

  1. [Isolation and diversity analyses of endophytic fungi from Paris polyphylla var. yunnanensis].

    PubMed

    Wang, Qian; Shen, Shi-Kang; Zhang, Ai-Li; Wu, Chun-Yan; Wu, Fu-Qin; Zhang, Xin-Jun; Wang, Yue-Hua

    2013-11-01

    The paper is aimed at studying the diversity of endophytic fungi community from Paris polyphylla var. yunnanensis, and to provide a scientific basis for the utilization value of the endophytic fungi as bioactive material resources. In the present study, endophytic fungi were isolated from roots, rhizomes and leaves of wild P. polyphylla var. yunnanensis collected from Baoshan, Heqing county and Songming city of Yunnan province, and identified and classified by morphological methods together with its ITS sequence analysis. Seven and forty-nine strains of endophytic fungi were isolated from P. polyphylla var. yunnanensis. They were identified belonging to 41 genus. In these 41 genus, 3 genus exist in root only, 12 genus only exist in rhizome and 8 genus only exist in leaf. There was difference in endophytic fungi isolated from different sample sites. Endophytic fungi diversity from rhizomes of Heqing site was the highest. Endophytic fungi similarity coefficient was low among different sites and tissues. Based on these results, it is reasonable to propose that endophytic fungi of P. polyphylla var. yannanensis from different tissue and different sample sites has a certain difference which is possibly relate to their different habitats, different structure and composition of each tissue.

  2. Molecular diversity of fungal endophytes isolated from Garcinia mangostana and Garcinia parvifolia.

    PubMed

    Sim, Jiun-Horng; Khoo, Chai-Hoon; Lee, Learn-Han; Cheah, Yoke-Kqueen

    2010-04-01

    Garcinia is commonly found in Malaysia, but limited information is available regarding endophytic fungi associated with this plant. In this study, 24 endophytic fungi were successfully recovered from different parts of two Garcinia species. Characterization of endophytic fungi was performed based on the conserved internal transcribed spacer (ITS) region sequence analysis and the antimicrobial properties. Results revealed that fruits of the plant appeared to be the highest inhabitation site (38 %) as compared with others. Glomerella sp., Guignardia sp., and Phomopsis sp. appeared to be the predominant endophytic fungi group in Garcinia mangostana and Garcinia parvifolia. Phylogenetic relationships of the isolated endophytic fungi were estimated from the sequences of the ITS region. On the other hand, antibacterial screening showed 11 of the isolates possessed positive response towards pathogenic and nonpathogenic bacteria. However, there was no direct association between certain antibacterial properties with the specific genus observed.

  3. Lignin-solubilizing ability of actinomycetes isolated from termite (Termitidae) gut. [Streptomyces viridosporus

    SciTech Connect

    Pasti, M.B.; Crawford, D.L. ); Pometto, A.L., III ); Nuti, M.P. )

    1990-07-01

    The lignocellulose-degrading abilities of 11 novel actinomycete strains isolated from termite gut were determined and compared with that of the well-characterized actinomycete, Streptomyces viridosporus T7A. Lignocellulose bioconversion was followed by (i) monitoring the degradation of ({sup 14}C)lignin- and ({sup 14}C)cellulose-labeled phloem of Abies concolor to {sup 14}CO{sub 2} and {sup 14}C-labeled water-soluble products, (ii) determining lignocellulose, lignin, and carbohydrate losses resulting from growth on a lignocellulose substrate prepared from corn stalks (Zea mays), and (iii) quantifying production of a water-soluble lignin degradation intermediate (acid-precipitable polymeric lignin). Of the assays used, total lignocellulose weight loss was most useful in determining overall bioconversion ability but not in identifying the best lignin-solubilizing strains. A screening procedure based on {sup 14}CO{sub 2} evolution from ({sup 14}C-lignin)lignocellulose combined with measurement of acid-precipitable polymeric lignin yield was the most effective in identifying lignin-solubilizing strains. For the termite gut strains, the pH of the medium showed no increase after 3 weeks of growth on lignocellulose. This is markedly different from the pattern observed with S. viridosporus T7A, which raises the medium pH considerably. Production of extracellular peroxidases by the 11 strains and S. viridosporus T7A was followed for 5 days in liquid cultures. On the basis of an increase of specific peroxidase activity in the presence of lignocellulose in the medium, the actinomycetes could be placed into the same three groups.

  4. Lignin-solubilizing ability of actinomycetes isolated from termite (Termitidae) gut.

    PubMed Central

    Pasti, M B; Pometto, A L; Nuti, M P; Crawford, D L

    1990-01-01

    The lignocellulose-degrading abilities of 11 novel actinomycete strains isolated from termite gut were determined and compared with that of the well-characterized actinomycete, Streptomyces viridosporus T7A. Lignocellulose bioconversion was followed by (i) monitoring the degradation of [14C]lignin- and [14C]cellulose-labeled phloem of Abies concolor to 14CO2 and 14C-labeled water-soluble products, (ii) determining lignocellulose, lignin, and carbohydrate losses resulting from growth on a lignocellulose substrate prepared from corn stalks (Zea mays), and (iii) quantifying production of a water-soluble lignin degradation intermediate (acid-precipitable polymeric lignin). The actinomycetes were all Streptomyces strains and could be placed into three groups, including a group of five strains that appear superior to S. viridosporus T7A in lignocellulose-degrading ability, three strains of approximately equal ability, and three strains of lesser ability. Strain A2 was clearly the superior and most effective lignocellulose decomposer of those tested. Of the assays used, total lignocellulose weight loss was most useful in determining overall bioconversion ability but not in identifying the best lignin-solubilizing strains. A screening procedure based on 14CO2 evolution from [14C-lignin]lignocellulose combined with measurement of acid-precipitable polymeric lignin yield was the most effective in identifying lignin-solubilizing strains. For the termite gut strains, the pH of the medium showed no increase after 3 weeks of growth on lignocellulose. This is markedly different from the pattern observed with S. viridosporus T7A, which raises the medium pH considerably. Production of extracellular peroxidases by the 11 strains and S. viridosporus T7A was followed for 5 days in liquid cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2167628

  5. Isolation and functional characterization of bacterial endophytes from Carica papaya fruits.

    PubMed

    Krishnan, P; Bhat, R; Kush, A; Ravikumar, P

    2012-08-01

    To isolate and characterize the endophytes from papaya fruits and to determine the fermentative potential of the strains. Endophytes provide potential sources for novel natural products for the use in agriculture and nutrition. There is very limited information on isolation and characterization of bacterial endophytes from papaya. We describe isolation and characterization of eighteen endophytes of papaya fruit from four economically important papaya varieties viz 'Red lady', 'Solo', 'Coorg Honey' and 'Bangalore'. The phylogenetic analysis based on the 16S rRNA sequence revealed that isolated endophytes are genetically distinct and cluster as discrete clades in the dendrogram. The Bacillus species is a predominant bacterial endophyte across papaya varieties. The seeds and the endocarp of papaya fruits harbour Kocuria, Acinetobacter and Enterobacter species. The Staphylococcus species were detected in the fruit mesocarp of two papaya varieties used in the study. The endophytes isolated from papaya fruits were capable of producing extracellular enzymes like amylase, cellulase, pectinase and xylanase. Three isolates, Bacillus (PE-LR-1 and PE-LR-3) and Kocuria (PE-LR-2), were selected for fruit fermentation, and antioxidant potential of the fermented product was evaluated. PE-LR-3 fermented product has the free radical scavenging activity of 61·2% and a microbial cocktail of PE-LR-3 with Saccharomyces cerevisiae MTCC 2918 enhances the antioxidant potential to 75·7%. These findings suggest that different parts of papaya fruits harbour an array of bacterial endophytes that could be important agents in attributing the high nutritive status to the fruit and can serve as potent microbial cocktails for developing value-added fermented products of this important fruit. This study describes isolation of a bacterial endophyte from papaya fruit that is capable of improving the antioxidant potential of raw papaya after fermentation. No claim to Indian Government works Journal

  6. An endophytic fungus efficiently producing paclitaxel isolated from Taxus wallichiana var. mairei

    PubMed Central

    Zaiyou, Jian; Li, Meng; Xiqiao, Hu

    2017-01-01

    Abstract Paclitaxel is a medicinal ingredient with high anticancer activity and widely used in hospitals and clinics. In this study, we isolate endophytic fungi efficiently producing paclitaxel from yew for the purpose of paclitaxel manufacture. The bark of Taxus wallichiana var. mairei was surface sterilized and then inoculated in potato dextrose agar culture medium to isolate endophytic fungi. The paclitaxel in the fungal culture was extracted with mixture of chloroform and the same amount of methanol. The content of paclitaxel in the extract was determined and identified with LC-MS. The endophytic fungus efficiently producing paclitaxel was species identified with ITS rDNA and 26S D1/D2 rDNA sequencing. There were 528 endophytic fungal strains were isolated from the bark of T wallichiana var. mairei in total. There was only a strain efficiently producing paclitaxel in these endophytic fungi. The unique strain was identified as Phoma medicaginis. The paclitaxel contents in whole potato dextrose broth (PDB) culture, spent culture medium from this strain and that in dry mycelium is 1.215 mg/L, 0.936 mg/L, and 20 mg/kg, respectively. An endophytic fungus efficiently producing paclitaxel was isolated from T wallichiana var. mairei. This isolated endophytic fungus can be used as a producing strain for paclitaxel manufacture. PMID:28682896

  7. An endophytic fungus efficiently producing paclitaxel isolated from Taxus wallichiana var. mairei.

    PubMed

    Zaiyou, Jian; Li, Meng; Xiqiao, Hu

    2017-07-01

    Paclitaxel is a medicinal ingredient with high anticancer activity and widely used in hospitals and clinics. In this study, we isolate endophytic fungi efficiently producing paclitaxel from yew for the purpose of paclitaxel manufacture.The bark of Taxus wallichiana var. mairei was surface sterilized and then inoculated in potato dextrose agar culture medium to isolate endophytic fungi. The paclitaxel in the fungal culture was extracted with mixture of chloroform and the same amount of methanol. The content of paclitaxel in the extract was determined and identified with LC-MS. The endophytic fungus efficiently producing paclitaxel was species identified with ITS rDNA and 26S D1/D2 rDNA sequencing.There were 528 endophytic fungal strains were isolated from the bark of T wallichiana var. mairei in total. There was only a strain efficiently producing paclitaxel in these endophytic fungi. The unique strain was identified as Phoma medicaginis. The paclitaxel contents in whole potato dextrose broth (PDB) culture, spent culture medium from this strain and that in dry mycelium is 1.215 mg/L, 0.936 mg/L, and 20 mg/kg, respectively.An endophytic fungus efficiently producing paclitaxel was isolated from T wallichiana var. mairei. This isolated endophytic fungus can be used as a producing strain for paclitaxel manufacture.

  8. Diversity of cultivated endophytic bacteria from sugarcane: genetic and biochemical characterization of Burkholderia cepacia complex isolates.

    PubMed

    Mendes, Rodrigo; Pizzirani-Kleiner, Aline A; Araujo, Welington L; Raaijmakers, Jos M

    2007-11-01

    Bacteria were isolated from the rhizosphere and from inside the roots and stems of sugarcane plants grown in the field in Brazil. Endophytic bacteria were found in both the roots and the stems of sugarcane plants, with a significantly higher density in the roots. Many of the cultivated endophytic bacteria were shown to produce the plant growth hormone indoleacetic acid, and this trait was more frequently found among bacteria from the stem. 16S rRNA gene sequence analysis revealed that the selected isolates of the endophytic bacterial community of sugarcane belong to the genera of Burkholderia, Pantoea, Pseudomonas, and Microbacterium. Bacterial isolates belonging to the genus Burkholderia were the most predominant among the endophytic bacteria. Many of the Burkholderia isolates produced the antifungal metabolite pyrrolnitrin, and all were able to grow at 37 degrees C. Phylogenetic analyses of the 16S rRNA gene and recA gene sequences indicated that the endophytic Burkholderia isolates from sugarcane are closely related to clinical isolates of the Burkholderia cepacia complex and clustered with B. cenocepacia (gv. III) isolates from cystic fibrosis patients. These results suggest that isolates of the B. cepacia complex are an integral part of the endophytic bacterial community of sugarcane in Brazil and reinforce the hypothesis that plant-associated environments may act as a niche for putative opportunistic human pathogenic bacteria.

  9. Nocardia kroppenstedtii sp. nov., an actinomycete isolated from a lung transplant patient with a pulmonary infection.

    PubMed

    Jones, Amanda L; Fisher, Andrew J; Mahida, Rahul; Gould, Kate; Perry, John D; Hannan, Margaret M; Judge, Eoin P; Brown, Ros; Boagey, Kimberley; Goodfellow, Michael

    2014-03-01

    A novel actinomycete, strain N1286(T), isolated from a lung transplant patient with a pulmonary infection, was provisionally assigned to the genus Nocardia. The strain had chemotaxonomic and morphological properties typical of members of the genus Nocardia and formed a distinct phyletic line in the Nocardia 16S rRNA gene tree. Isolate N1286(T) was most closely related to Nocardia farcinica DSM 43665(T) (99.8% gene sequence similarity) but could be distinguished from the latter by the low level of DNA-DNA relatedness. These strains were also distinguishable on the basis of a broad range of phenotypic properties. It is concluded that strain N1286(T) represents a novel species of the genus Nocardia for which the name Nocardia kroppenstedtii sp. nov. is proposed. The type strain is N1286(T) ( = DSM 45810(T) = NCTC 13617(T)).

  10. Isolation of Amylolytic Strains of Thermoactinomyces vulgaris and Production of Thermophilic Actinomycete Amylases1

    PubMed Central

    Kuo, M. J.; Hartman, P. A.

    1966-01-01

    Kuo, M. J. (Iowa State University, Ames), and P. A. Hartman. Isolation of amylolytic strains of Thermoactinomyces vulgaris and production of thermophilic actinomycete amylases. J. Bacteriol. 92:723–726. 1966.—Of 759 isolates obtained from dung, compost, and soil samples, a culture of Thermoactinomyces vulgaris (strain 5) was selected for further study on the basis of quantities of amylase produced in synthetic and nonsynthetic media, rapid growth and sporulation, culture stability upon prolonged storage at 5 C, and growth temperature range. Inoculum preparation, temperature optimum for amylase formation, and the effects of various kinds and levels of carbon and nitrogen sources on amylase production were studied with T. vulgaris strain 5. An optimal procedure for production of T. vulgaris amylases is proposed. PMID:5922543

  11. Isolation and identification of bacterial endophytes from pharmaceutical agarwood-producing Aquilaria species

    PubMed Central

    Bhore, Subhash J.; Preveena, Jagadesan; Kandasamy, Kodi I.

    2013-01-01

    Background: Resins and gums are used in traditional medicine and do have potential applications in pharmacy and medicine. Agarwood is the fragrant resinous wood, which is an important commodity from Aquilaria species and has been used as a sedative, analgesic, and digestive in traditional medicine. Endophytic bacteria are potentially important in producing pharmaceutical compounds found in the plants. Hence, it was important to understand which types of endophytic bacteria are associated with pharmaceutical agarwood-producing Aquilaria species. Objective: This study was undertaken to isolate and identify endophytic bacteria associated with agarwood-producing seven (7) Aquilaria species from Malaysia. Materials and Methods: Botanical samples of seven Aquilaria species were collected, and endophytic bacteria were isolated from surface-sterilized-tissue samples. The 16S rRNA gene fragments were amplified using PCR method, and endophytic bacterial isolates (EBIs) were identified based on 16S rRNA gene sequence similarity based method. Results: Culturable, 77 EBIs were analyzed, and results of 16S rRNA gene sequences analysis suggest that 18 different types of endophytic bacteria are associated with (seven) Aquilaria species. From 77 EBIs, majority (36.4%) of the isolates were of Bacillus pumilus. Conclusion: These findings indicate that agarwood-producing Aquilaria species are harboring 18 different types of culturable endophytic bacteria. PMID:23798890

  12. Antimicrobial Activities of Some Actinomycetes Isolated from Different Rhizospheric Soils in Tunisia.

    PubMed

    Trabelsi, Ines; Oves, Daniel; Manteca, Angel; Genilloud, Olga; Altalhi, Abdullah; Nour, Mohamed

    2016-08-01

    Fifty four isolates of actinomycetes were collected from four different rhizospheric soils: 18 strains from palm tree bark and soil, 12 strains from an olive field soil, 9 strains from a coastal forest, and 15 strains from an agriculture soil situated in the Algerian-Tunisian border (Oum Tboul). Based on morphological and cultural characters, the isolates were classified as Streptomyces (42 strains), Micromonospora (4 strains), Pseudonocardia (1 strain), Actinomadura (1 strain), Nocardia (1 strain), and non-Streptomyces (5 strains). More than half of the isolates inhibited at least one tested pathogenic microorganisms in liquid culture. In addition, antimicrobial activities of some strains were tested on solid culture. Several bioactive compounds were identified by liquid chromatography joined with low-resolution mass spectroscopy (LC/MS) and analysed by MEDINA's database and by the dictionary of natural products Chapman & Hall. An interesting chlorinated compound with the molecular formula C20H37ClN2O4, produced by three different strains (SF1, SF2, and SF5), was subject of an attempted purification. However, it was demonstrated using confocal microscopy and LC/MS high resolution that this compound is produced only on solid culture. These three potential antimicrobial isolates showed high similarity with Streptomyces thinghirensis and Streptomyces lienomycini, in terms of morphological characteristics and 16S rRNA gene sequences (bootstrap 97 %). All these findings prove the high antimicrobial diversity of the studied soils. The potential of the selected and other relatively unexplored extreme environments constitute a source of interesting actinomycete strains producing several biologically active secondary metabolites.

  13. Plant growth-promoting potential of endophytic fungi isolated from Solanum nigrum leaves.

    PubMed

    Khan, Abdur Rahim; Ullah, Ihsan; Waqas, Muhammad; Shahzad, Raheem; Hong, Sung-Jun; Park, Gun-Seok; Jung, Byung Kwon; Lee, In-Jung; Shin, Jae-Ho

    2015-09-01

    Fungal endophytes have been characterized as producers of phytohormones and potent promoters of plant growth. In this study, two fungal endophytes, Fusarium tricinctum RSF-4L and Alternaria alternata RSF-6L, were isolated from the leaves of Solanum nigrum. Culture filtrates (CFs) from each isolate were initially screened for indole compounds, and assayed for their ability to promote the growth of Dongjin rice plants. Nearly all plant growth attributes examined (i.e., chlorophyll content, root-shoot length, and biomass production) were significantly enhanced upon treatment with fungal CFs. Subsequently, gas chromatography/mass spectrometry analyses were utilized to confirm the presence of phytohormones in the CF of each fungal endophytic isolate. These analyses revealed that RSF-4L and RSF-6L produced 54 and 30 µg/mL indole acetic acid, respectively, within their respective cultures. These findings suggest that the endophytes isolated in this study synthesize bioactive compounds that could play important roles in promoting plant growth.

  14. In vitro interaction of actinomycetes isolates with Aspergillus flavus: impact on aflatoxins B1 and B2 production.

    PubMed

    Verheecke, C; Liboz, T; Darriet, M; Sabaou, N; Mathieu, F

    2014-06-01

    This work aimed to study the interaction between Actinomycetal isolates and Aspergillus flavus to promote mutual antagonism in contact. Thirty-seven soilborn Streptomyces spp. isolates were chosen as potential candidates. After a 10-day in vitro co-incubation period, 27 isolates respond to the criteria, that is, mutual antagonism in contact. Further aflatoxins B1 and B2 analysis revealed that those 27 isolates reduced aflatoxin B1 residual concentration from 38·6 to 4·4%, depending on the isolate. We selected 12 isolates and tested their capacity to reduce AFB1 in pure culture to start identifying the mechanisms involved in its reduction. AFB1 was reduced by eight isolates. The remaining AFB1 concentration varied between 82·2 and 15·6%. These findings led us to suggest that these eight isolates could be used as biocontrol agents against AFB1 and B2 with low risk of impacting the natural microbial equilibrium. Interaction between Aspergillus flavus and Actinomycetes isolates was conducted in vitro. Actinomycetes isolates having a mutual antagonism in contact with A. flavus were chosen for further aflatoxins production study. This is a new approach based to develop biocontrol against aflatoxins accumulation in maize while respecting natural microbial equilibrium. © 2014 The Society for Applied Microbiology.

  15. Identification and characterization of endophytic bacteria isolated from in vitro cultures of peach and pear rootstocks.

    PubMed

    Liaqat, Fakhra; Eltem, Rengin

    2016-12-01

    Endophytes are microorganisms which live symbiotically with almost all varieties of plant and in turn helping the plant in a number of ways. Instead of satisfactory surface sterilization approaches, repeatedly occurring bacterial growth on in vitro rootstock cultures of peach and pear was identified and isolated as endophytic bacteria in our present study. Five different isolates from peach rootstocks were molecularly identified by 16S rRNA gene sequencing as Brevundimonas diminuta, Leifsonia shinshuensis, Sphingomonas parapaucimobilis Brevundimonas vesicularis, Agrobacterium tumefaciens while two endophytic isolates of pear were identified as Pseudoxanthomonas mexicana, and Stenotrophomonas rhizophilia. Identified endophytes were also screened for their potential of plant growth promotion according to indoleacetic acid (IAA) production, nitrogen fixation, solubilization of phosphate and production of siderophore. All seven endophytic isolates have shown positive results for IAA, nitrogen fixation and phosphate solubilization tests. However, two out of seven isolates showed positive results for siderophore production. On the basis of these growth promoting competences, isolated endophytes can be presumed to have significant influence on the growth of host plants. Future studies required to determine the antimicrobial susceptibility profile and potential application of these isolates in biological control, microbial biofertilizers and degradative enzyme production.

  16. Actinopolyspora algeriensis sp. nov., a novel halophilic actinomycete isolated from a Saharan soil.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2012-09-01

    A halophilic actinomycete strain designated H19(T), was isolated from a Saharan soil in the Bamendil region (Ouargla province, South Algeria) and was characterized taxonomically by using a polyphasic approach. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Actinopolyspora, and 16S rRNA gene sequence analysis confirmed that strain H19(T) was a novel species of the genus Actinopolyspora. DNA-DNA hybridization value between strain H19(T) and the nearest Actinopolyspora species, A. halophila, was clearly below the 70 % threshold. The genotypic and phenotypic data showed that the organism represents a novel species of the genus Actinopolyspora for which the name Actinopolyspora algeriensis sp. nov. is proposed, with the type strain H19(T) (= DSM 45476(T) = CCUG 62415(T)).

  17. Pseudonocardia hispaniensis sp. nov., a novel actinomycete isolated from industrial wastewater activated sludge.

    PubMed

    Cuesta, G; Soler, A; Alonso, J L; Ruvira, M A; Lucena, T; Arahal, D R; Goodfellow, M

    2013-01-01

    A novel actinomycete, designated PA3(T), was isolated from an oil refinery wastewater treatment plant, located in Palos de la Frontera, Huelva, Spain, and characterized taxonomically by using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate formed a distinct subclade in the Pseudonocardia tree together with Pseudonocardia asaccharolytica DSM 44247(T). The chemotaxonomic properties of the isolate, for example, the presence of MK-8 (H(4)) as the predominant menaquinone and iso-C(16:0) as the major fatty acid, are consistent with its classification in the genus Pseudonocardia. DNA:DNA pairing experiments between the isolate and the type strain of P. asaccharolytica DSM 44247(T) showed that they belonged to separate genomic species. The two strains were readily distinguished using a combination of phenotypic properties. Consequently, it is proposed that isolate PA3(T) represents a novel species for which the name Pseudonocardia hispaniensis sp. nov. is proposed. The type strain is PA3(T) (= CCM 8391(T) = CECT 8030(T)).

  18. Isolation and characterization of indigenous endophytic bacteria associated with leaves of switchgrass (Panicum virgatum L.) cultivars.

    PubMed

    Gagne-Bourgue, F; Aliferis, K A; Seguin, P; Rani, M; Samson, R; Jabaji, S

    2013-03-01

    To isolate and characterize indigenous bacterial endophytes from cultivars of switchgrass and study their antimicrobial and growth promoting potential. The diversity, molecular and biochemical characterizations of indigenous and culturable bacterial endophytes residing in leaves of switchgrass have not been studied previously. This study describes the characterization of 31 bacterial endophytes from three switchgrass cutlivars: Cave-in Rock, Blue Jacket and Tecumseh. Molecular and phylogenetic analysis based on the 16S rRNA sequence grouped the endophytes into eight different taxa that shared high homology of 98-99% with other known sequences. Bacterial endophytes were identified as Microbacterium testaceum, Curtobacterium flaccumfaciens, Bacillus subtilis and Bacillus pumilus, Pseudomonas fluorescens, Sphingomonas parapaucimobilis, Serratia sp. and Pantoea ananatis. Some endophytes were detected in switchgrass seeds and in plants that originated from seeds collected a year earlier, confirming vertical transmission to the next generation of the host. Selected endophytes produced cellulases and were capable of solubilizing inorganic phosphorus. Analysis of cell-free culture filtrate of selected strains using direct infusion orbitrap mass spectrometry confirmed the presence of several well-characterized lipopeptide toxins and phytohormones. Re-inoculation of the roots of switchgrass seedlings with endophytes singly or combined confirmed their migration to the upper aerial parts of the plant. Our findings suggest that switchgrass leaves harbour a diversity of bacterial endophytes, some of which could potentially be applied as growth promoting bacteria. This is the first report on the characterization of indigenous bacterial endophytes and their potential use as biofertilizers. © 2012 The Society for Applied Microbiology.

  19. Transformation of 2,4,6-trinitrotoluene (TNT) by actinomycetes isolated from TNT-contaminated and uncontaminated environments

    SciTech Connect

    Pasti-Grigsby, M.B.; Lewis, T.A.; Crawford, D.L.; Crawford, R.L.

    1996-03-01

    Biotransformation of TNT has been reported under both aerobic and anaerobic conditions. Actinomycetes are important decomposers in composts. This study examines the tolerance of acitomycete cultures, isolated from both TNT-contaminated and uncontaminated environments for different concentrations to TNT, determined how selected isolates transform TNT, and examined whether such TNT transformations were constitutive or induced by exposure to TNT. 33 refs., 1 figs., 1 tab.

  20. Characterization of Five Fungal Endophytes Producing Cajaninstilbene Acid Isolated from Pigeon Pea [Cajanus cajan (L.) Millsp.

    PubMed Central

    Zu, Yuan Gang; Fu, Yu Jie; Wang, Wei; Luo, Meng; Efferth, Thomas

    2011-01-01

    Five fungal endophytes (K4, K5, K6, K9, K14) producing Cajaninstilbene acid (CSA, 3-hydroxy-4-prenyl-5-methoxystilbene-2-carboxylic acid) were isolated from the roots of pigeon pea [Cajanus cajan (L.) Millsp.]. CSA is responsible for the prominent pharmacological activities in pigeon pea. The amount of CSA in culture solution varied among the five fungal endophytes. K4 produced the highest levels of CSA (1037.13 µg/L) among the endophytes tested after incubation for five days. Both morphological characteristics and molecular methods were used for species identification of fungal endophytes. The five endophytic isolates were characterized by analyzing the internal transcribed spacer (ITS) rRNA and β-tubulin genes. The K4, K5, K9 and K14 strains isolated from pigeon pea roots were found to be closely related to the species Fusarium oxysporum. K6 was identified as Neonectria macrodidym. The present study is the first report on the isolation and identification of fungal endophytes producing CSA in pigeon pea. The study also provides a scientific base for large scale production of CSA. PMID:22102911

  1. A method to type the potential angucycline producers in actinomycetes isolated from marine sponges.

    PubMed

    Ouyang, Yongchang; Wu, Houbo; Xie, Lianwu; Wang, Guanghua; Dai, Shikun; Chen, Minjie; Yang, Keqian; Li, Xiang

    2011-05-01

    Angucyclines are aromatic polyketides with antimicrobial, antitumor, antiviral and enzyme inhibition activities. In this study, a new pair of degenerate primers targeting the cyclase genes that are involved in the aromatization of the first and/or second ring of angucycline, were designed and evaluated in a PCR protocol targeting the jadomycin cyclase gene of Streptomyces venezuelae ISP5230. The identity of the target amplicon was confirmed by sequencing. After validation, the primers were used to screen 49 actinomycete isolates from three different marine sponges to identify putative angucycline producers. Seven isolates were positively identified using this method. Sequence analysis of the positive amplicons confirmed their identity as putative angucycline cyclases with sequence highly similar to known angucycline cyclases. Phylogenetic analysis clustered these positives into the angucycline group of cyclases. Furthermore, amplifications of the seven isolates using ketosynthase-specific primers were positive, backing the results using the cyclase primers. Together these results provided strong support for the presence of angucycline biosynthetic genes in these isolates. The specific primer set targeting the cyclase can be used to identify putative angucycline producers among marine actinobacteria, and aid in the discovery of novel angucyclines.

  2. Isolation, identification and bioactivity of endophytic fungi from medicinal plant Malus sieboldii.

    PubMed

    Cai, Guanghua; Wang, Xiaoling

    2012-03-01

    To isolate and identify endophytic fungi from Malus sieboldii, and detect cytotoxicity, protease inhibition and antifungal activities of their crude extracts. The fungi were identified with the aid of morphology or Internal Transcribed Spacer (ITS) rDNA molecular methods. Fungal activities were tested by cylinder-plate, MTT and BRpNA methods, respectively. A total of 217 endophytic fungi were isolated from M. sieboldii. Of the 22 taxa obtained, non-sporulating, Alternaria, Colletotrichum, Aspergillu, Fusarlum, Gliocladium and Cunninghamella were dominant communities. The result of the bioactivity test showed that 30 endophytic fungi displayed inhibition against at least one pathogenic fungus, and 3 and 4 showed cytotoxicity and protease inhibition, respectively. M. sieboldii should be a potential source of bioactive endophytic fungi.

  3. Culturable bacterial endophytes isolated from Mangrove tree (Rhizophora apiculata Blume) enhance seedling growth in Rice

    PubMed Central

    Deivanai, Subramanian; Bindusara, Amitraghata Santhanam; Prabhakaran, Guruswamy; Bhore, Subhash Janardhan

    2014-01-01

    Background: Endophytic bacteria do have several potential applications in medicine and in other various sectors of biotechnology including agriculture. Bacterial endophytes need to be explored for their potential applications in agricultural biotechnology. One of the potential applications of bacterial endophytes in agricultural is to enhance the growth of the agricultural crops. Hence, this study was undertaken to explore the plant growth promoting potential application of bacterial endophytes. Objective: The objective of this study was to examine the effect of endophytic bacteria from mangrove tree (Rhizophora apiculata Blume) for their efficacy in promoting seedling growth in rice. Materials and Methods: Eight endophytic bacterial isolates (EBIs) isolated from twig and petiole tissues of the mangrove were identified based on their 16S ribosomal ribonucleic acid (rRNA) gene sequence homology. Separately, surface sterilized paddy seeds were treated with cell-free broth and cell suspension of the EBIs. Rice seedlings were analyzed by various bioassays and data was recorded. Results: The gene sequences of the isolates were closely related to two genera namely, Bacillus and Pantoea. Inoculation of EBIs from R. apiculata with rice seeds resulted in accelerated root and shoot growth with significant increase in chlorophyll content. Among the isolates, Pantoea ananatis (1MSE1) and Bacillus amyloliquefaciens (3MPE1) had shown predominance of activity. Endophytic invasion was recognized by the non-host by rapid accumulation of reactive oxygen species (ROS) and was counteracted by the production of hydrogen peroxide (H2O2) and lipid peroxide. The results demonstrated that EBIs from mangrove tree can increase the fitness of the rice seedlings under controlled conditions. Conclusion: These research findings could be useful to enhance the seedling growth and could serve as foundation in further research on enhancing the growth of the rice crop using endophytic bacteria. PMID

  4. Molecular Identification of Endophytic Fungi Isolated from Needle Leaves of Conifers in Bohyeon Mountain, Korea

    PubMed Central

    Yoo, Jae-Joon

    2012-01-01

    Fungal endophytes are microfungi that live in plants without causing apparent symptoms of infection. This study was conducted to identify endophytic fungi isolated from leaves of coniferous trees in Bohyeon Mountain of Korea. We collected leaves of two species of coniferous trees, Pinus densiflora and Pinus koraiensis, from 11 sites in the study area. A total 58 isolates were obtained and identified using molecular and morphological characteristics. Four species of endophytic fungi were isolated from P. densiflora: Lophodermium conigenum, Leotiomycetes sp., Septoria pini-thunbergii, and Polyporales sp., while two fungal species were isolated from P. koraiensis: Eurotiomycetes sp. and Rhytismataceae sp. The most frequently isolated species were L. conigenum and S. pini-thunbergii. PMID:23323047

  5. Molecular identification of endophytic fungi isolated from needle leaves of conifers in bohyeon mountain, Korea.

    PubMed

    Yoo, Jae-Joon; Eom, Ahn-Heum

    2012-12-01

    Fungal endophytes are microfungi that live in plants without causing apparent symptoms of infection. This study was conducted to identify endophytic fungi isolated from leaves of coniferous trees in Bohyeon Mountain of Korea. We collected leaves of two species of coniferous trees, Pinus densiflora and Pinus koraiensis, from 11 sites in the study area. A total 58 isolates were obtained and identified using molecular and morphological characteristics. Four species of endophytic fungi were isolated from P. densiflora: Lophodermium conigenum, Leotiomycetes sp., Septoria pini-thunbergii, and Polyporales sp., while two fungal species were isolated from P. koraiensis: Eurotiomycetes sp. and Rhytismataceae sp. The most frequently isolated species were L. conigenum and S. pini-thunbergii.

  6. Extracellular enzymatic profiles and taxonomic identification of endophytic fungi isolated from four plant species.

    PubMed

    Alberto, R N; Costa, A T; Polonio, J C; Santos, M S; Rhoden, S A; Azevedo, J L; Pamphile, J A

    2016-11-03

    Plants of medicinal and economic importance have been studied to investigate the presence of enzyme-producing endophytic fungi. The characterization of isolates with distinct enzyme production potential may identify suitable alternatives for specialized industry. At Universidade Estadual de Maringá Laboratory of Microbial Biotechnology, approximately 500 isolates of endophytic fungi have been studied over the last decade from various host plants, including medicinally and economically important species, such as Luehea divaricata (Martius et Zuccarini), Trichilia elegans A. Juss, Sapindus saponaria L., Piper hispidum Swartz, and Saccharum spp. However, only a fraction of these endophytes have been identified and evaluated for their biotechnological application, having been initially grouped by morphological characteristics, with at least one representative of each morphogroup tested. In the current study, several fungal strains from four plants (L. divaricata, T. elegans, S. saponaria, and Saccharum spp) were identified by ribosomal DNA typing and evaluated semi-quantitatively for their enzymatic properties, including amylase, cellulase, pectinase, and protease activity. Phylogenetic analysis revealed the presence of four genera of endophytic fungi (Diaporthe, Saccharicola, Bipolaris, and Phoma) in the plants examined. According to enzymatic tests, 62% of the isolates exhibited amylase, approximately 93% cellulase, 50% pectinase, and 64% protease activity. Our results verified that the composition and abundance of endophytic fungi differed between the plants tested, and that these endophytes are a potential enzyme production resource of commercial and biotechnological value.

  7. Antifungal and proteolytic activities of endophytic fungi isolated from Piper hispidum Sw.

    PubMed

    Orlandelli, Ravely Casarotti; de Almeida, Tiago Tognolli; Alberto, Raiani Nascimento; Polonio, Julio Cesar; Azevedo, João Lúcio; Pamphile, João Alencar

    2015-06-01

    Endophytes are being considered for use in biological control, and the enzymes they secrete might facilitate their initial colonization of internal plant tissues and direct interactions with microbial pathogens. Microbial proteases are also biotechnologically important products employed in bioremediation processes, cosmetics, and the pharmaceutical, photographic and food industries. In the present study, we evaluated antagonism and competitive interactions between 98 fungal endophytes and Alternaria alternata, Colletotrichum sp., Phyllosticta citricarpa and Moniliophthora perniciosa. We also examined the proteolytic activities of endophytes grown in liquid medium and conducted cup plate assays. The results showed that certain strains in the assemblage of P. hispidum endophytes are important sources of antifungal properties, primarily Lasiodiplodia theobromae JF766989, which reduced phytopathogen growth by approximately 54 to 65%. We detected 28 endophytes producing enzymatic halos of up to 16.40 mm in diameter. The results obtained in the present study highlight the proteolytic activity of the endophytes Phoma herbarum JF766995 and Schizophyllum commune JF766994, which presented the highest enzymatic halo diameters under at least one culture condition tested. The increased activities of certain isolates in the presence of rice or soy flour as a substrate (with halos up to 17.67 mm in diameter) suggests that these endophytes have the potential to produce enzymes using agricultural wastes.

  8. Antifungal and proteolytic activities of endophytic fungi isolated from Piper hispidum Sw

    PubMed Central

    Orlandelli, Ravely Casarotti; de Almeida, Tiago Tognolli; Alberto, Raiani Nascimento; Polonio, Julio Cesar; Azevedo, João Lúcio; Pamphile, João Alencar

    2015-01-01

    Endophytes are being considered for use in biological control, and the enzymes they secrete might facilitate their initial colonization of internal plant tissues and direct interactions with microbial pathogens. Microbial proteases are also biotechnologically important products employed in bioremediation processes, cosmetics, and the pharmaceutical, photographic and food industries. In the present study, we evaluated antagonism and competitive interactions between 98 fungal endophytes and Alternaria alternata, Colletotrichum sp., Phyllosticta citricarpa and Moniliophthora perniciosa. We also examined the proteolytic activities of endophytes grown in liquid medium and conducted cup plate assays. The results showed that certain strains in the assemblage of P. hispidum endophytes are important sources of antifungal properties, primarily Lasiodiplodia theobromae JF766989, which reduced phytopathogen growth by approximately 54 to 65%. We detected 28 endophytes producing enzymatic halos of up to 16.40 mm in diameter. The results obtained in the present study highlight the proteolytic activity of the endophytes Phoma herbarum JF766995 and Schizophyllum commune JF766994, which presented the highest enzymatic halo diameters under at least one culture condition tested. The increased activities of certain isolates in the presence of rice or soy flour as a substrate (with halos up to 17.67 mm in diameter) suggests that these endophytes have the potential to produce enzymes using agricultural wastes. PMID:26273250

  9. In Vitro Evaluation of Enzymatic and Antifungal Activities of Soil-Actinomycetes Isolates and Their Molecular Identification by PCR

    PubMed Central

    Keikha, Nasser; Ayatollahi Mousavi, Seyyed Amin; Nakhaei, Ali Reza; Yadegari, Mohammad Hossein; Shahidi Bonjar, Gholam Hossein; Amiri, Somayyeh

    2015-01-01

    Background: Human cutaneous infection caused by a homogeneous group of keratinophilic fungi called dermatophytes. These fungi are the most common infectious agents in humans that are free of any population and geographic area. Microsporum canis is a cause of dermatophytosis (Tinea) in recent years in Iran and atypical strain has been isolated in Iran. Its cases occur sporadically due to M. canis transmission from puppies and cats to humans. Since this pathogenic dermatophyte is eukaryotes, chemical treatment with antifungal drugs may also affect host tissue cells. Objectives: The aim of the current study was to find a new antifungal agent of soil-Actinomycetes from Kerman province against M. canis and Actinomycete isolates were identified by PCR. Materials and Methods: A number of hundred Actinomycete isolated strains were evaluated from soil of Kerman province, for their antagonistic activity against the M. canis. M. canis of the Persian Type Culture Collection (PTCC) was obtained from the Iranian Research Organization for Science and Technology (IROST). Electron microscope studies of these isolates were performed based on the physiological properties of these antagonists including lipase, amylase, protease and chitinase activities according to the relevant protocols and were identified using gene 16SrDNA. Results: In this study the most antagonist of Actinomycete isolates with antifungal activity against M. canis isolates of L1, D5, Ks1m, Km2, Kn1, Ks8 and Ks1 were shown in vitro. Electron microscopic studies showed that some fungal strains form spores, mycelia and spore chain. Nucleotide analysis showed that Ks8 had maximum homology (98%) to Streptomyces zaomyceticus strain xsd08149 and L1 displayed 100% homology to Streptomyces sp. HVG6 using 16SrDNA studies. Conclusions: Our findings showed that Streptomyces has antifungal effects against M. canis. PMID:26060560

  10. Glycomyces tarimensis sp. nov., an actinomycete isolated from a saline-alkali habitat.

    PubMed

    Lv, Ling-Ling; Zhang, Yue-Feng; Zhang, Li-Li

    2015-05-01

    A novel actinomycete strain, designated TRM 45387(T), was isolated from a saline-alkali soil in Xinjiang Province (40° 22' N 79° 08' E), north-west China. The isolate was characterized using a polyphasic approach. 16S rRNA gene sequence analysis indicated that strain TRM 45387(T) belonged to the genus Glycomyces and was closely related to Glycomyces arizonensis DSM 44726(T) (96.59% 16S rRNA gene sequence similarity). The G+C content of the DNA was 71.26 mol%. The isolate contained meso-diaminopimelic acid as the diagnostic diamino acid, and xylose, glucose, galactose, arabinose and ribose as the major whole-cell sugars. The diagnostic phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides. The predominant menaquinone was MK-10(H6). The major fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. On the basis of the evidence from this polyphasic study, a novel species, Glycomyces tarimensis sp. nov., is proposed. The type strain of Glycomyces tarimensis is TRM 45387(T) ( =CCTCC AA 2014007(T) =JCM 30184(T)). © 2015 Xinjiang Production & Construction Corps Key Laborartory of Protection and Utilization of Biological Resources in Tarim Basin.

  11. Amycolatopsis nigrescens sp. nov., an actinomycete isolated from a Roman catacomb.

    PubMed

    Groth, Ingrid; Tan, Geok Yuan Annie; González, Juan M; Laiz, Leonila; Carlsohn, Marc René; Schütze, Barbara; Wink, Joachim; Goodfellow, Michael

    2007-03-01

    The taxonomic status of two actinomycetes isolated from the wall of a hypogean Roman catacomb was established based on a polyphasic investigation. The organisms were found to have chemical and morphological markers typical of members of the genus Amycolatopsis. They also shared a range of chemical, molecular and phenotypic markers which served to separate them from representatives of recognized Amycolatopsis species. The new isolates formed a branch in the Amycolatopsis 16S rRNA gene sequence tree with Amycolatopsis minnesotensis NRRL B-24435(T), but this association was not supported by a particularly high bootstrap value or by the product of the maximum-parsimony tree-making algorithm. The organisms were distinguished readily from closely related Amycolatopsis species based on a combination of phenotypic properties and from all Amycolatopsis strains by their characteristic menaquinone profiles, in which tetra-hydrogenated menaquinones with 11 isoprene units predominated. The combined genotypic and phenotypic data indicate that the isolates merit recognition as representing a novel species of the genus Amycolatopsis. The name proposed for this novel species is Amycolatopsis nigrescens sp. nov., with type strain CSC17Ta-90(T) (=HKI 0330(T)=DSM 44992(T)=NRRL B-24473(T)).

  12. Streptomyces gamaensis sp. nov., a novel actinomycete with antifungal activity isolated from soil in Gama, Chad.

    PubMed

    Zhao, Shanshan; Ye, Lan; Liu, Chongxi; Abagana, Adam Yacoub; Zheng, Weiwei; Sun, Pengyu; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing

    2017-04-01

    During an investigation exploring potential sources of novel species and natural products, a novel actinomycete with antifungal activity, designated strain NEAU-Gz11(T), was isolated from a soil sample, which was collected from Gama, Chad. The isolate was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain NEAU-Gz11(T) belongs to the genus Streptomyces with high sequence similarity to Streptomyces hiroshimensis JCM 4098(T) (98.0 %). Similarities to other type strains of the genus Streptomyces were lower than 98.0 %. However, the physiological and biochemical characteristics and low levels of DNA-DNA relatedness could differentiate the isolate genotypically and phenotypically from S. hiroshimensis JCM 4098(T). Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces gamaensis sp. nov. is proposed. The type strain is NEAU-Gz11(T) (=CGMCC 4.7304(T)=DSM 101531(T)).

  13. Streptomyces atlanticus sp. nov., a novel actinomycete isolated from marine sponge Aplysina fulva (Pallas, 1766).

    PubMed

    Silva, Fábio Sérgio Paulino; Souza, Danilo Tosta; Zucchi, Tiago Domingues; Pansa, Camila Cristiane; de Figueiredo Vasconcellos, Rafael Leandro; Crevelin, Eduardo José; de Moraes, Luiz Alberto Beraldo; Melo, Itamar Soares

    2016-11-01

    The taxonomic position of a novel marine actinomycete isolated from a marine sponge, Aplysina fulva, which had been collected in the Archipelago of Saint Peter and Saint Paul (Equatorial Atlantic Ocean), was determined by using a polyphasic approach. The organism showed a combination of morphological and chemotaxonomic characteristics consistent with its classification in the genus Streptomyces and forms a distinct branch within the Streptomyces somaliensis 16S rRNA gene tree subclade. It is closely related to Streptomyces violascens ISP 5183(T) (97.27 % 16S rRNA gene sequence similarity) and Streptomyces hydrogenans NBRC 13475(T) (97.15 % 16S rRNA gene sequence similarity). The 16S rRNA gene similarities between the isolate and the remaining members of the subclade are lower than 96.77 %. The organism can be distinguished readily from other members of the S. violacens subclade using a combination of phenotypic properties. On the basis of these results, it is proposed that isolate 103(T) (=NRRL B-65309(T) = CMAA 1378(T)) merits recognition as the type strain of a new Streptomyces species, namely Streptomyces atlanticus sp. nov.

  14. Actinoplanes hulinensis sp. nov., a novel actinomycete isolated from soybean root (Glycine max (L.) Merr).

    PubMed

    Shen, Yue; Liu, Chongxi; Wang, Xiangjing; Zhao, Junwei; Jia, Feiyu; Zhang, Yuqin; Wang, Liang; Yang, Deguang; Xiang, Wensheng

    2013-02-01

    A novel actinomycete, designated strain NEAU-M9(T), was isolated from soybean root (Glycine max (L.) Merr) and characterized using a polyphasic approach. 16S rRNA gene sequence similarity studies showed that strain NEAU-M9(T) belonged to the genus Actinoplanes, being most closely related to Actinoplanes campanulatus DSM 43148(T) (98.85 %), Actinoplanes capillaceus DSM 44859(T) (98.70 %), Actinoplanes lobatus DSM 43150(T) (98.30 %), Actinoplanes auranticolor DSM 43031(T) (98.23 %) and Actinoplanes sichuanensis 03-723(T) (98.06 %); similarity to other type strains of the genus Actinoplanes ranged from 95.87 to 97.56 %. The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that the isolate formed a distinct phyletic line with A. campanulatus DSM 43148(T) and A. capillaceus DSM 44859(T). This branching pattern was also supported by the tree constructed with the maximum-likelihood method. However, the low level of DNA-DNA relatedness allowed the isolate to be differentiated from the above-mentioned two Actinoplanes species. Moreover, strain NEAU-M9(T) could also be distinguished from the most closely related species by morphological, physiological and characteristics. Therefore, it is proposed that strain NEAU-M9(T) represents a novel Actinoplanes species, Actinoplanes hulinensis sp. nov. The type strain of Actinoplanes hulinensis is NEAU-M9(T) (= CGMCC 4.7036(T) = DSM 45728(T)).

  15. Field studies on two rock phosphate solubilizing actinomycete isolates as biofertilizer sources

    NASA Astrophysics Data System (ADS)

    Mba, Caroline C.

    1994-03-01

    Recently biotechnology is focusing attention on utilization of biological resources to solve a number of environmental problems such as soil fertility management. Results of microbial studies on earthworm compost in the University of Nigeria farm identified a number of rock phosphate solubilizing actinomycetes. Two of these, isclates 02 and 13, were found to be efficient rock phosphate (RP) solubilizers and fast-growing cellulolytic microbes producing extracellular hydrolase enzymes. In this preliminary field study the two microbial isolates were investigated with respect to their effects on the growth of soybean and egusi as well as their effect on the incidence of toxicity of poultry droppings. Application of these isolates in poultry manure-treated field plots, as microbial fertilizers, brought about yield increases of 43% and 17% with soybeans and 19% and 33% with egusi, respectively. Soil properties were also improved. With isolates 02 and 13, the soil available phosphorus increased at the five-leaf stage, while N-fixation in the soil increased by 45% or 11% relative to control. It was further observed that air-dried poultry manure after four days of incubation was still toxic to soybean. The toxic effect of the applied poultry manure was reduced or eliminated with microbial fertilizers 02 or 13, respectively. The beneficial effects of the microbial organic fertilizer are discussed. Justification for more intensive research on rock phosphate organic fertilizer is highlighted.

  16. Isolation and enzyme bioprospection of endophytic bacteria associated with plants of Brazilian mangrove ecosystem.

    PubMed

    Castro, Renata A; Quecine, Maria Carolina; Lacava, Paulo T; Batista, Bruna D; Luvizotto, Danice M; Marcon, Joelma; Ferreira, Anderson; Melo, Itamar S; Azevedo, João L

    2014-01-01

    The mangrove ecosystem is a coastal tropical biome located in the transition zone between land and sea that is characterized by periodic flooding, which confers unique and specific environmental conditions on this biome. In these ecosystems, the vegetation is dominated by a particular group of plant species that provide a unique environment harboring diverse groups of microorganisms, including the endophytic microorganisms that are the focus of this study. Because of their intimate association with plants, endophytic microorganisms could be explored for biotechnologically significant products, such as enzymes, proteins, antibiotics and others. Here, we isolated endophytic microorganisms from two mangrove species, Rhizophora mangle and Avicennia nitida, that are found in streams in two mangrove systems in Bertioga and Cananéia, Brazil. Bacillus was the most frequently isolated genus, comprising 42% of the species isolated from Cananéia and 28% of the species from Bertioga. However, other common endophytic genera such as Pantoea, Curtobacterium and Enterobacter were also found. After identifying the isolates, the bacterial communities were evaluated for enzyme production. Protease activity was observed in 75% of the isolates, while endoglucanase activity occurred in 62% of the isolates. Bacillus showed the highest activity rates for amylase and esterase and endoglucanase. To our knowledge, this is the first reported diversity analysis performed on endophytic bacteria obtained from the branches of mangrove trees and the first overview of the specific enzymes produced by different bacterial genera. This work contributes to our knowledge of the microorganisms and enzymes present in mangrove ecosystems.

  17. Molecular phylogeny and diversity of Fusarium endophytes isolated from tomato stems.

    PubMed

    Imazaki, Iori; Kadota, Ikuo

    2015-09-01

    Plant tissues are a known habitat for two types of Fusarium species: plant pathogens and endophytes. Here, we investigated the molecular phylogeny and diversity of endophytic fusaria, because endophytes are not as well studied as pathogens. A total of 543 Fusarium isolates were obtained from the inside of tomato stems cultivated in soils mainly obtained from agricultural fields. We then determined partial nucleotide sequences of the translation elongation factor-1 alpha (EF-1α) genes of the isolates. Among the isolates from tomato, 24 EF-1α gene sequence types (EFST) were found: nine were classified as being from the Fusarium oxysporum species complex and its sister taxa (FOSC, 332 isolates), seven from the F. fujikuroi species complex (FFSC, 75 isolates) and eight from the F. solani species complex (FSSC, 136 isolates). To determine more characteristic details of the tomato isolates, we isolated 180 fusaria directly from soils and found 95% of them were nested within the FOSC (82 isolates; five EFSTs), FFSC (21 isolates; six FESTs) and FSSC (68 isolates; 11 EFSTs). These results suggested that the dominant Fusarium endophytes within tomato stems were members of the same three species complexes, which were also the dominant fusaria in the soils.

  18. Spatial and temporal variation in fungal endophyte communities isolated from cultivated cotton (Gossypium hirsutum).

    PubMed

    Ek-Ramos, María J; Zhou, Wenqing; Valencia, César U; Antwi, Josephine B; Kalns, Lauren L; Morgan, Gaylon D; Kerns, David L; Sword, Gregory A

    2013-01-01

    Studies of fungi in upland cotton (Gossypium hirsutum) cultivated in the United States have largely focused on monitoring and controlling plant pathogens. Given increasing interest in asymptomatic fungal endophytes as potential biological control agents, surveys are needed to better characterize their diversity, distribution patterns and possible applications in integrated pest management. We sampled multiple varieties of cotton in Texas, USA and tested for temporal and spatial variation in fungal endophyte diversity and community composition, as well as for differences associated with organic and conventional farming practices. Fungal isolates were identified by morphological and DNA identification methods. We found members of the genera Alternaria, Colletotrichum and Phomopsis, previously isolated as endophytes from other plant species. Other recovered species such as Drechslerella dactyloides (formerly Arthrobotrys dactyloides) and Exserohilum rostratum have not, to our knowledge, been previously reported as endophytes in cotton. We also isolated many latent pathogens, but some species such as Alternaria tennuissima, Epicoccum nigrum, Acremonium alternatum, Cladosporium cladosporioides, Chaetomium globosum and Paecilomyces sp., are known to be antagonists against plant pathogens, insects and nematode pests. We found no differences in endophyte species richness or diversity among different cotton varieties, but did detect differences over time and in different plant tissues. No consistent patterns of community similarity associated with variety, region, farming practice, time of the season or tissue type were observed regardless of the ecological community similarity measurements used. Results indicated that local fungal endophyte communities may be affected by both time of the year and plant tissue, but the specific community composition varies across sites. In addition to providing insights into fungal endophyte community structure, our survey provides

  19. Spatial and Temporal Variation in Fungal Endophyte Communities Isolated from Cultivated Cotton (Gossypium hirsutum)

    PubMed Central

    Ek-Ramos, María J.; Zhou, Wenqing; Valencia, César U.; Antwi, Josephine B.; Kalns, Lauren L.; Morgan, Gaylon D.; Kerns, David L.; Sword, Gregory A.

    2013-01-01

    Studies of fungi in upland cotton (Gossypium hirsutum) cultivated in the United States have largely focused on monitoring and controlling plant pathogens. Given increasing interest in asymptomatic fungal endophytes as potential biological control agents, surveys are needed to better characterize their diversity, distribution patterns and possible applications in integrated pest management. We sampled multiple varieties of cotton in Texas, USA and tested for temporal and spatial variation in fungal endophyte diversity and community composition, as well as for differences associated with organic and conventional farming practices. Fungal isolates were identified by morphological and DNA identification methods. We found members of the genera Alternaria, Colletotrichum and Phomopsis, previously isolated as endophytes from other plant species. Other recovered species such as Drechslerella dactyloides (formerly Arthrobotrys dactyloides) and Exserohilum rostratum have not, to our knowledge, been previously reported as endophytes in cotton. We also isolated many latent pathogens, but some species such as Alternaria tennuissima, Epicoccum nigrum, Acremonium alternatum, Cladosporium cladosporioides, Chaetomium globosum and Paecilomyces sp., are known to be antagonists against plant pathogens, insects and nematode pests. We found no differences in endophyte species richness or diversity among different cotton varieties, but did detect differences over time and in different plant tissues. No consistent patterns of community similarity associated with variety, region, farming practice, time of the season or tissue type were observed regardless of the ecological community similarity measurements used. Results indicated that local fungal endophyte communities may be affected by both time of the year and plant tissue, but the specific community composition varies across sites. In addition to providing insights into fungal endophyte community structure, our survey provides

  20. Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination

    PubMed Central

    Lumactud, Rhea; Shen, Shu Yi; Lau, Mimas; Fulthorpe, Roberta

    2016-01-01

    The bacterial endophytic communities of four plants growing abundantly in soils highly contaminated by hydrocarbons were analyzed through culturable and culture-independent means. Given their tolerance to the high levels of petroleum contamination at our study site, we sought evidence that Achillea millefolium, Solidago canadensis, Trifolium aureum, and Dactylis glomerata support high levels of hydrocarbon degrading endophytes. A total of 190 isolates were isolated from four plant species. The isolates were identified by partial 16S rDNA sequence analysis, with class Actinobacteria as the dominant group in all species except S. canadensis, which was dominated by Gammaproteobacteria. Microbacterium foliorum and Plantibacter flavus were present in all the plants, with M. foliorum showing predominance in D. glomerata and both endophytic bacterial species dominated T. aureum. More than 50% of the isolates demonstrated degradative capabilities for octanol, toluene, naphthalene, kerosene, or motor oil based on sole carbon source growth screens involving the reduction of tetrazolium dye. P. flavus isolates from all the sampled plants showed growth on all the petroleum hydrocarbons (PHCs) substrates tested. Mineralization of toluene and naphthalene was confirmed using gas-chromatography. 16S based terminal restriction fragment length polymorphism analysis revealed significant differences between the endophytic bacterial communities showing them to be plant host specific at this site. To our knowledge, this is the first account of the degradation potential of bacterial endophytes in these commonly occurring pioneer plants that were not previously known as phytoremediating plants. PMID:27252685

  1. Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination.

    PubMed

    Lumactud, Rhea; Shen, Shu Yi; Lau, Mimas; Fulthorpe, Roberta

    2016-01-01

    The bacterial endophytic communities of four plants growing abundantly in soils highly contaminated by hydrocarbons were analyzed through culturable and culture-independent means. Given their tolerance to the high levels of petroleum contamination at our study site, we sought evidence that Achillea millefolium, Solidago canadensis, Trifolium aureum, and Dactylis glomerata support high levels of hydrocarbon degrading endophytes. A total of 190 isolates were isolated from four plant species. The isolates were identified by partial 16S rDNA sequence analysis, with class Actinobacteria as the dominant group in all species except S. canadensis, which was dominated by Gammaproteobacteria. Microbacterium foliorum and Plantibacter flavus were present in all the plants, with M. foliorum showing predominance in D. glomerata and both endophytic bacterial species dominated T. aureum. More than 50% of the isolates demonstrated degradative capabilities for octanol, toluene, naphthalene, kerosene, or motor oil based on sole carbon source growth screens involving the reduction of tetrazolium dye. P. flavus isolates from all the sampled plants showed growth on all the petroleum hydrocarbons (PHCs) substrates tested. Mineralization of toluene and naphthalene was confirmed using gas-chromatography. 16S based terminal restriction fragment length polymorphism analysis revealed significant differences between the endophytic bacterial communities showing them to be plant host specific at this site. To our knowledge, this is the first account of the degradation potential of bacterial endophytes in these commonly occurring pioneer plants that were not previously known as phytoremediating plants.

  2. Amycolatopsis thailandensis sp. nov., a poly(L-lactic acid)-degrading actinomycete, isolated from soil.

    PubMed

    Chomchoei, Atchareeya; Pathom-Aree, Wasu; Yokota, Akira; Kanongnuch, Chartchai; Lumyong, Saisamorn

    2011-04-01

    A novel actinomycete that was capable of degrading poly(l-lactic acid), strain CMU-PLA07(T), was isolated from soil in northern Thailand. Strain CMU-PLA07(T) had biochemical, chemotaxonomic, morphological and physiological properties that were consistent with its classification in the genus Amycolatopsis. 16S rRNA gene sequence analysis showed that the isolate formed a phyletic line within the genus Amycolatopsis. Strain CMU-PLA07(T) was most similar to Amycolatopsis coloradensis IMSNU 22096(T) (99.5 % 16S rRNA gene sequence similarity) and Amycolatopsis alba DSM 44262(T) (99.4 %). However, strain CMU-PLA07(T) was distinguishable from the type strains of species of the genus Amycolatopsis on the basis of DNA-DNA relatedness and phenotypic data. Therefore, strain CMU-PLA07(T) is considered to represent a novel species of the genus Amycolatopsis, for which the name Amycolatopsis thailandensis sp. nov. is proposed. The type strain is CMU-PLA07(T) ( = JCM 16380(T) = BCC 38279(T)).

  3. Nocardiopsis arabia sp. nov., a halotolerant actinomycete isolated from a sand-dune soil.

    PubMed

    Hozzein, Wael N; Goodfellow, Michael

    2008-11-01

    The taxonomic status of an unknown actinomycete isolated from a sand-dune soil was established using a polyphasic approach. Isolate S186(T) had chemotaxonomic and morphological properties consistent with its classification in the genus Nocardiopsis, grew on agar plates at NaCl concentrations of up to 15 % (w/v) and formed a distinct phyletic line in the Nocardiopsis 16S rRNA gene sequence tree. Its closest phylogenetic neighbours were Nocardiopsis chromatogenes, Nocardiopsis composta, Nocardiopsis gilva and Nocardiopsis trehalosi, with sequence similarity to the various type strains of 96.9 %, but it was readily distinguished from the type strains of these and related species using a range of phenotypic properties. It is apparent from the genotypic and phenotypic data that strain S186(T) belongs to a novel species of the genus Nocardiopsis, for which the name Nocardiopsis arabia sp. nov. is proposed. The type strain is S186(T) (=CGMCC 4.2057(T) =DSM 45083(T)).

  4. Actinorugispora endophytica gen. nov., sp. nov., an actinomycete isolated from Daucus carota.

    PubMed

    Liu, Min-Jiao; Zhu, Wen-Yong; Li, Jie; Zhao, Guo-Zhen; Xiong, Zhi; Park, Dong-Jin; Hozzein, Wael N; Kim, Chang-Jin; Li, Wen-Jun

    2015-08-01

    An actinomycete strain, designated YIM 690008T, was isolated from Daucus carota collected from South Korea and its taxonomic position was investigated by using a polyphasic approach. The strain grew well on most media tested and no diffusible pigment was produced. The aerial mycelium formed wrinkled single spores and short spore chains, some of which were branched. The whole-cell hydrolysates contained meso-diaminopimelic acid, glucose, mannose, ribose, galactose and rhamnose. The predominant menaquinones were MK-10(H4), MK-10(H6), MK-10(H8) and MK-10(H2). The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannosides, some unknown phospholipids, glycolipids and polar lipids. The major fatty acids were i-C16 : 0, ai-C17 : 0 and C18 : 1ω9c. The DNA G+C content of the genomic DNA was 63.1 mol%. Phylogenetic analysis indicated that the isolate belongs to the family Nocardiopsaceae. However, based on phenotypic, chemotaxonomic and genotypic data, it was concluded that strain YIM 690008T represents a novel genus and novel species of the family Nocardiopsaceae, for which the name Actinorugispora endophytica gen. nov., sp. nov. (type strain YIM 690008T = DSM 46770T = JCM 30099T = KCTC 29480T) is proposed.

  5. Actinomycetes Diversity among rRNA Gene Clones and Cellular Isolates from Sambhar Salt Lake, India

    PubMed Central

    Yadav, A. K.; Vardhan, S.; Kashyap, S.; Yandigeri, M.; Arora, D. K.

    2013-01-01

    The vertical stratification of actinomycetes diversity in Sambhar salt lake (India's largest salt lake) was investigated by using cultivable and uncultivable approaches. The isolates from cultured approaches were clustered on the basis of cultural, morphological, biochemical, and cell wall characteristics, and results were further strengthened by 16S rDNA-RFLP into five major groups. 16S rDNA sequencing of the representative isolates from each clusters was identified as belonging to Streptomyces, Actinopolyspora, Microbispora, Saccharopolyspora, and Actinoplanes genera, while culture independent group was established as Streptomyces (130 clones, 20 OTUs), Micromonospora (96 clones, 7 OTUs), Streptosporangium (79 clones, 9 OTUs), Thermomonospora (46 clones, 8 OTUs), and Dactylosporangium (58 clones, 8 OTUs). The diversity assessment using Shannon and Wiener index was found to be 1.55, 1.52, 1.55, and 1.49 from surface lake water, at depth of 1.5 m, shallow layer of water with algal population, and finally at depth of 2.5 m, respectively. We observed diversity in terms of the species richness as Streptomyces is dominant genus in both culture dependent and culture independent techniques followed by Microbispora (culture dependent methods) and Micromonospora (culture independent method) genera, respectively. PMID:24307879

  6. Streptomyces kronopolitis sp. nov., an actinomycete that produces phoslactomycins isolated from a millipede (Kronopolites svenhedind Verhoeff).

    PubMed

    Liu, Chongxi; Ye, Lan; Li, Yao; Jiang, Shanwen; Liu, Hui; Yan, Kai; Xiang, Wensheng; Wang, Xiangjing

    2016-12-01

    A phoslactomycin-producing actinomycete, designated strain NEAU-ML8T, was isolated from a millipede (Kronopolites svenhedind Verhoeff) and characterized using a polyphasic approach. 16S rRNA gene sequence analysis showed that strain NEAU-ML8T belongs to the genus Streptomyces with the highest sequence similarities to Streptomyces lydicus NBRC 13058T (99.39 %) and Streptomyces chattanoogensis DSM 40002T (99.25 %). The maximum-likelihood phylogenetic tree based on 16S rRNA gene sequences showed that the isolate formed a distinct phyletic line with NBRC 13058T and S. chattanoogensis DSM 40002T. This branching pattern was also supported by the tree rconstructed with the neighbour-joining method. A combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain NEAU-ML8T and its phylogenetically closely related strains, which further clarified their relatedness and demonstrated that NEAU-ML8T could be distinguished from NBRC 13058T and S. chattanoogensis DSM 40002T. Therefore, it is concluded that strain NEAU-ML8T can be classified as representing a novel species of the genus Streptomyces, for which the name Streptomyces kronopolitis sp. nov. is proposed. The type strain is NEAU-ML8T (=DSM 101986T=CGMCC 4.7323T).

  7. Saccharopolyspora spongiae sp. nov., a novel actinomycete isolated from the marine sponge Scopalina ruetzleri (Wiedenmayer, 1977).

    PubMed

    Souza, Danilo Tosta; Silva, Fábio Sérgio Paulino da; Silva, Leonardo José da; Crevelin, Eduardo José; Moraes, Luiz Alberto Beraldo; Zucchi, Tiago Domingues; Melo, Itamar Soares

    2017-06-01

    A novel marine actinomycete, designated strain CMAA 1452T, was isolated from the sponge Scopalina ruetzleri collected from Saint Peter and Saint Paul Archipelago, in Brazil, and subjected to a polyphasic taxonomic investigation. The organism formed a distinct phyletic line in the Saccharopolyspora 16S rRNA gene tree and had chemotaxonomic and morphological properties consistent with its classification in this genus. It was found to be closely related to Saccharopolyspora dendranthemae KLBMP 1305T (99.5% 16S rRNA gene sequence similarity) and shared similarities of 99.3, 99.2 and 99.0 % with 'Saccharopolyspora endophytica' YIM 61095, Saccharopolyspora tripterygii YIM 65359T and 'Saccharopolyspora pathumthaniensis' S582, respectively. DNA-DNA relatedness values between the isolate and its closest phylogenetic neighbours, namely S. dendranthemae KLBMP 1305T, 'S. endophytica' YIM 61095 and S. tripterygii YIM 65359T, were 53.5, 25.8 and 53.2 %, respectively. Strain CMAA 1452T was also distinguished from the type strains of these species using a range of phenotypic features. On the basis of these results, it is proposed that strain CMAA 1452T (=DSM 103218T=NRRL B-65384T) merits recognition as the type strain of a novel Saccharopolyspora species, Saccharopolyspora spongiae sp. nov.

  8. Plantactinospora veratri sp. nov., an actinomycete isolated from black false hellebore root (Veratrum nigrum L.).

    PubMed

    Xing, He; Liu, Chongxi; Zhang, Yuejing; Zhao, Junwei; Li, Chuang; Liu, Hui; Li, Lianjie; Wang, Xiangjing; Xiang, Wensheng

    2015-06-01

    A novel actinomycete, designated strain NEAU-FHS4T, was isolated from the root of black false hellebore (Veratrum nigrum L.). Strain NEAU-FHS4T formed single spores with smooth surfaces on substrate mycelium. The novel strain contained meso-diaminopimelic as amino acid of the peptidoglycan and xylose and glucose as whole-cell sugars. The predominant menaquinones were MK-10(H6) and MK-10(H8). Mycolic acids were not detected. The diagnostic phospholipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol. The predominant cellular fatty acids were iso-C16 : 0, C16 : 0, C18 : 0 and anteiso-C17 : 0. Phenotypic and chemotaxonomic analysis showed that the novel isolate had characteristics typical of members of the genus Plantactinospora. 16S rRNA gene sequence analysis also indicated that strain NEAU-FHS4T belonged to the genus Plantactinospora, with highest sequence similarities to Plantactinospora mayteni YIM 61359T (98.88 %) and Plantactinospora endophytica YIM 68255T (98.85 %). The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the novel strain from the most closely related strains. Based on morphological, chemotaxonomic and phylogenetic data, strain NEAU-FHS4T is considered to represent a novel species of the genus Plantactinospora, for which the name Plantactinospora veratri sp. nov. is proposed. The type strain is NEAU-FHS4T ( = CGMCC 4.7143T = DSM 46718T).

  9. Diversity and seasonal variation of endophytic fungi isolated from three conifers in mt. Taehwa, Korea.

    PubMed

    Kim, Chang-Kyun; Eo, Ju-Kyeong; Eom, Ahn-Heum

    2013-06-01

    The needled leaves of three conifer species were collected in Mt. Taehwa during different seasons of the year. Total 59 isolates and 19 species of endophytic fungi were isolated from the leaves and identified using morphological and molecular characteristics. As a result, Shannon index was different in its host plant; Larix kaempferi had a highest value of species diversity. According to the sampling season, 9 species of 19 species were isolated during fall season. The results suggest that the existing of host plant and sampling season are major factors of distribution of endophytic fungi.

  10. Diversity and Seasonal Variation of Endophytic Fungi Isolated from Three Conifers in Mt. Taehwa, Korea

    PubMed Central

    Kim, Chang-Kyun; Eo, Ju-Kyeong

    2013-01-01

    The needled leaves of three conifer species were collected in Mt. Taehwa during different seasons of the year. Total 59 isolates and 19 species of endophytic fungi were isolated from the leaves and identified using morphological and molecular characteristics. As a result, Shannon index was different in its host plant; Larix kaempferi had a highest value of species diversity. According to the sampling season, 9 species of 19 species were isolated during fall season. The results suggest that the existing of host plant and sampling season are major factors of distribution of endophytic fungi. PMID:23874130

  11. Isolation and Characterization of Endophytic Colonizing Bacteria from Agronomic Crops and Prairie Plants†

    PubMed Central

    Zinniel, Denise K.; Lambrecht, Pat; Harris, N. Beth; Feng, Zhengyu; Kuczmarski, Daniel; Higley, Phyllis; Ishimaru, Carol A.; Arunakumari, Alahari; Barletta, Raúl G.; Vidaver, Anne K.

    2002-01-01

    Endophytic bacteria reside within plant hosts without causing disease symptoms. In this study, 853 endophytic strains were isolated from aerial tissues of four agronomic crop species and 27 prairie plant species. We determined several phenotypic properties and found approximately equal numbers of gram-negative and gram-positive isolates. In a greenhouse study, 28 of 86 prairie plant endophytes were found to colonize their original hosts at 42 days postinoculation at levels of 3.5 to 7.7 log10 CFU/g (fresh weight). More comprehensive colonization studies were conducted with 373 corn and sorghum endophytes. In growth room studies, none of the isolates displayed pathogenicity, and 69 of the strains were recovered from corn or sorghum seedlings at levels of 8.3 log10 CFU/plant or higher. Host range greenhouse studies demonstrated that 26 of 29 endophytes were recoverable from at least one host other than corn and sorghum at levels of up to 5.8 log10 CFU/g (fresh weight). Long-range dent corn greenhouse studies and field trials with 17 wild-type strains and 14 antibiotic-resistant mutants demonstrated bacterial persistence at significant average colonization levels ranging between 3.4 and 6.1 log10 CFU/g (fresh weight) up to 78 days postinoculation. Three prairie and three agronomic endophytes exhibiting the most promising levels of colonization and an ability to persist were identified as Cellulomonas, Clavibacter, Curtobacterium, and Microbacterium isolates by 16S rRNA gene sequence, fatty acid, and carbon source utilization analyses. This study defines for the first time the endophytic nature of Microbacterium testaceum. These microorganisms may be useful for biocontrol and other applications. PMID:11976089

  12. Draft genome sequences of three chemically rich actinomycetes isolated from Mediterranean sponges.

    PubMed

    Horn, Hannes; Cheng, Cheng; Edrada-Ebel, RuAngelie; Hentschel, Ute; Abdelmohsen, Usama Ramadan

    2015-12-01

    Metabolomic analysis has shown the chemical richness of the sponge-associated actinomycetes Streptomyces sp. SBT349, Nonomureae sp. SBT364, and Nocardiopsis sp. SBT366. The genomes of these actinomycetes were sequenced and the genomic potential for secondary metabolism was evaluated. Their draft genomes have sizes of 8.0, 10, and 5.8 Mb having 687, 367, and 179 contigs with a GC content of 71.6, 70.7, and 72.7%, respectively. Moreover, antiSMASH 3.0 predicted 108, 149, and 75 secondary metabolite gene clusters, respectively which highlight the metabolic capacity of the three actinomycete species to produce diverse classes of natural products.

  13. ISOLATION AND IDENTIFICATION OF AN ENDOPHYTIC FUNGUS PRODUCING PACLITAXEL FROM TAXUS WALLICHIANA VAR MAIREI.

    PubMed

    Zaiyou, Jian; Hongsheng, Wang; Ning, Wang; Li, Meng; Guifang, Xu

    2015-12-01

    The objective of this study was to isolate endophytic fungi producing paclitaxel from yew for the purpose of paclitaxel manufacture. Surface sterilized bark of Taxus wallichiana var. mairei was used as source material and potato dextrose agar culture medium was used in isolation of endophytic fungi. Fungal cultures were extracted with a mixture of chloroform / methanol (1:1, v/v) and the paclitaxel in the extracts was determined and authenticated with LC-MS. An endophytic fungus that produced paclitaxel was identified by ITS rDNA and 26S D1/D2 rDNA sequencing. The results showed that a total of 435 endophytic fungal strains were isolated from T. wallichiana var. mairei and purified. Only one of these strains produced paclitaxel and it belongs to Fusarium. The paclitaxel productivity in whole PDB culture and that in spent culture medium from this strain is 0.0153 mg/L and 0.0119 mg/L respectively. The paclitaxel content in dry mycelium is 0.27 mg/kg. This isolated endophytic fungus produced paclitaxel at a considerable level and shows potentiality as a producing strain for paclitaxel manufacture after strain improvement. Copyright AULA MEDICA EDICIONES 2014. Published by AULA MEDICA. All rights reserved.

  14. Nonomuraea solani sp. nov., an actinomycete isolated from eggplant root (Solanum melongena L.).

    PubMed

    Wang, Xiangjing; Zhao, Junwei; Liu, Chongxi; Wang, Jidong; Shen, Yue; Jia, Feiyu; Wang, Liang; Zhang, Ji; Yu, Chao; Xiang, Wensheng

    2013-07-01

    A novel actinomycete, designated strain NEAU-Z6(T), was isolated from eggplant (Solanum melongena L.) root. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain NEAU-Z6(T) belonged to the genus Nonomuraea, with highest sequence similarity to Nonomuraea monospora PT 708(T) (98.83 %), Nonomuraea rosea GW 12687(T) (98.55 %) and Nonomuraea rhizophila YIM 67092(T) (98.02 %). Sequence similarities between strain NEAU-Z6(T) and other species of the genus Nonomuraea ranged from 97.94 % (Nonomuraea candida HMC10(T)) to 96.30 % (Nonomuraea wenchangensis 210417(T)). Key morphological, physiological and chemotaxonomic characteristics of strain NEAU-Z6(T) were congruent with the description of the genus Nonomuraea. The G+C content of the genomic DNA was 64.51 mol%. DNA-DNA relatedness and comparative analysis of physiological, biochemical and chemotaxonomic data allowed genotypic and phenotypic differentiation of strain NEAU-Z6(T) from closely related species. Thus, strain NEAU-Z6(T) represents a novel species of the genus Nonomuraea, for which the name Nonomuraea solani sp. nov. is proposed. The type strain is NEAU-Z6(T) ( = CGMCC 4.7037(T) = DSM 45729(T)).

  15. Demetria terragena gen. nov., sp. nov., a new genus of actinomycetes isolated from compost soil.

    PubMed

    Groth, I; Schumann, P; Rainey, F A; Martin, K; Schuetze, B; Augsten, K

    1997-10-01

    A novel actinomycete was isolated from compost soil and was studied taxonomically and phylogenetically. Cells of this organism were gram positive, not acid fast, nonmotile, nonsporulating, irregular coccoid to short rod shaped, and microaerophilic. The cell wall peptidoglycan contained lysine and was cross-linked via an L-Lys<--L-Ser<--D-Asp interpeptide bridge. The major menaquinone was MK-8(H4). The polar lipids were phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, and two unknown phospholipids. Mycolic acids were absent. The cellular fatty acid profile was complex, with large amounts of saturated and monounsaturated straight-chain acids and smaller amounts of iso and anteiso branched-chain acids. The G+C content of the DNA was 66 mol%. Comparative 16S ribosomal DNA studies revealed that strain HKI 0089T represents a novel lineage within Actinobacteria (32) distinct from all previously described genera and most closely related to members of the genera Kytococcus, Dermacoccus, and Dermatophilus of the family Dermatophilaceae. On the basis of our results, we suggest that strain HKI 0089 should be classified in a new genus and species, for which we propose the name Demetria terragena. The type strain and the only strain of the genus and species is HKI 0089 (DSM 11295).

  16. Actinomycetospora iriomotensis sp. nov., a novel actinomycete isolated from a lichen sample.

    PubMed

    Yamamura, Hideki; Ashizawa, Haruna; Nakagawa, Youji; Hamada, Moriyuki; Ishida, Yuumi; Otoguro, Misa; Tamura, Tomohiko; Hayakawa, Masayuki

    2011-04-01

    An actinomycete strain, IR73-Li102(T), was isolated from a lichen sample obtained from Iriomote Island, Japan, and subsequently characterized using a polyphasic approach. Comparative 16S rRNA gene sequence analysis revealed that strain IR73-Li102(T) had the highest sequence similarities with Actinomycetospora chiangmaiensis YIM 0006(T) (98.3%), A. corticola 014-5(T) (98.1%) and A. rishiriensis RI109-Li102(T) (98.0%). However, DNA-DNA hybridization assays, as well as physiological and biochemical analyzes, showed that strain IR73-Li102(T) could be clearly differentiated from its closest phylogenetic relatives. The strain contained meso-diaminopimelic acid, and arabinose and galactose were present in whole-cell hydrolysates. The predominant menaquinone was MK-8(H(4)), and the diagnostic phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and diphosphatidylglycerol. The major cellular fatty acid was iso-C(16:0) (58%). The chemotaxonomic properties of strain IR73-Li102(T) were consistent with those shared by members of the genus Actinomycetospora. On the basis of the phenotypic features and DNA-DNA hybridization data, strain IR73-Li102(T) (= NBRC 106365(T) = KCTC 19783(T)) represents a novel species of the genus Actinomycetospora, for which the name Actinomycetospora iriomotensis sp. nov. is proposed.

  17. Streptomyces castaneus sp. nov., a novel actinomycete isolated from the rhizosphere of Peucedanum praeruptorum Dunn.

    PubMed

    Zhou, Shuyu; Li, Zhilei; Bai, Lu; Yan, Kai; Zhao, Junwei; Lu, Chang; Liu, Chongxi; Wang, Xiangjing; Xiang, Wensheng

    2017-01-01

    During an investigation of microbial diversity in medicinal herbs, a novel actinomycete, strain NEAU-QHHV11(T) was isolated from the rhizosphere of Peucedanum praeruptorum Dunn collected from Xianglu Mountain in Heilongjiang Province, northeast China and characterized using a polyphasic approach. The organism was found to have typical characteristics of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequence also indicated that strain NEAU-QHHV11(T) belongs to the genus Streptomyces and was most closely related to Streptomyces graminilatus NBRC 108882(T) (98.7 % sequence similarity) and Streptomyces turgidiscabies NBRC 16080(T) (98.7 % sequence similarity). The results of DNA-DNA hybridization and some phenotypic characteristics indicated that strain NEAU-QHHV11(T) could be distinguished from its close phylogenetic relatives. Thus, strain NEAU-QHHV11(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces castaneus sp. nov. is proposed. The type strain is NEAU-QHHV11(T) (=CGMCC 4.7235(T) = DSM 100520(T)).

  18. Actinomadura gamaensis sp. nov., a novel actinomycete isolated from soil in Gama, Chad.

    PubMed

    Abagana, Adam Yacoub; Sun, Pengyu; Liu, Chongxi; Cao, Tingting; Zheng, Weiwei; Zhao, Shanshan; Xiang, Wensheng; Wang, Xiangjing

    2016-06-01

    A novel single spore-producing actinomycete, designated strain NEAU-Gz5(T), was isolated from a soil sample from Gama, Chad. A polyphasic taxonomic study was carried out to establish the status of this strain. The diamino acid present in the cell wall is meso-diaminopimelic acid. Glucose, mannose and madurose occur in whole cell hydrolysates. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside and an unidentified glycolipid. The predominant menaquinones were identified as MK-9(H8) and MK-9(H6). The predominant cellular fatty acids were found to be C16:0, iso-C15:0, iso-C16:0 and C18:0 10-methyl. Phylogenetic analysis based on the 16S rRNA gene showed that strain NEAU-Gz5(T) belongs to the genus Actinomadura and is closely related to Actinomadura oligospora JCM 10648(T) (ATCC 43269(T); 98.3 % similarity). However, the low level of DNA-DNA relatedness and some different phenotypic characteristics allowed the strain to be distinguished from its close relatives. Therefore, it is concluded that strain NEAU-Gz5(T) represents a novel species of the genus of Actinomadura, for which the name Actinomadura gamaensis sp. nov. is proposed. The type strain is NEAU-Gz5(T) (= CGMCC 4.7301(T) = DSM 100815(T)).

  19. Actinopolyspora mzabensis sp. nov., a halophilic actinomycete isolated from an Algerian Saharan soil.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-10-01

    A halophilic actinomycete strain, designated H55(T), was isolated from Saharan soil sampled in the Mzab region (Ghardaïa, southern Algeria) and was characterized in a taxonomic study using a polyphasic approach. The cell wall was determined to contain meso-diaminopimelic acid and the characteristic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-10(H4) and MK-9(H4). The predominant cellular fatty acids were determined to be anteiso-C17 : 0, iso-C16 : 0 and iso-C15 : 0. The diagnostic phospholipid detected was phosphatidylcholine. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Actinopolyspora, and 16S rRNA gene sequence analysis confirmed that strain H55(T) was a member of this genus. DNA-DNA hybridization values between strain H55(T) and the type strains of the nearest species of the genus Actinopolyspora, Actinopolyspora erythraea and A. alba, were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the organism represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora mzabensis sp. nov. is proposed, with the type strain H55(T) ( = DSM 45460(T) = CCUG 62965(T)).

  20. Saccharothrix saharensis sp. nov., an actinomycete isolated from Algerian Saharan soil.

    PubMed

    Boubetra, Dalila; Zitouni, Abdelghani; Bouras, Noureddine; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-10-01

    The taxonomic position of a novel actinomycete, strain SA152(T), isolated from a sample of Algerian Saharan soil, was determined using a polyphasic taxonomic approach. The strain produced abundant aerial mycelium and fragmented substrate mycelium on most media tested. Chemotaxonomically and phylogenetically, the strain was related to the members of the genus Saccharothrix. Results of 16S rRNA gene sequence comparison revealed that strain SA152(T) shared the highest degree of 16S rRNA gene sequence similarity with Saccharothrix xinjiangensis NBRC 101911(T) (99.3 %) and Saccharothrix texasensis NRRL B-16134(T) (98.9 %). However, DNA-DNA hybridization studies showed only 16.2 % relatedness with S. xinjiangensis DSM 44896(T) and 33.9 % relatedness with S. texasensis DSM 44231(T). Based upon genotypic and phenotypic differences from other members of the genus, a novel species, Saccharothrix saharensis sp. nov., is proposed, with SA152(T) ( = DSM 45456(T) = CCUG 60213(T)) as the type strain.

  1. Prauserella isguenensis sp. nov., a halophilic actinomycete isolated from desert soil.

    PubMed

    Saker, Rafika; Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-05-01

    Two actinomycete strains, designated H225(T) and H137, were isolated from two soil samples collected from the arid region of Ahbas at Béni-Isguen (Mzab), located in the Algerian Sahara. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the novel strains should be assigned to the genus Prauserella of the family Pseudonocardiaceae , and they were therefore subjected to a polyphasic taxonomic study. These two strains contained meso-diaminopimelic acid as the diagnostic diamino acid and arabinose and galactose as major whole-cell sugars. The diagnostic phospholipid was phosphatidylethanolamine. The predominant menaquinone was MK-9(H4), and the major fatty acid was iso-C16 : 0. DNA-DNA hybridization values between strain H225(T) and its closest phylogenetic neighbours, namely Prauserella flava DSM 45265(T), Prauserella alba DSM 44590(T), Prauserella aidingensis DSM 45266(T), Prauserella salsuginis DSM 45264(T) and Prauserella sediminis DSM 45267(T), were clearly below the 70% threshold used for species delineation. The genomic DNA G+C content of strains H225(T) and H137 was 70.4 mol%. On the basis of phenotypic and genotypic data, strains H225(T) and H137(T) are considered to represent a novel species of the genus Prauserella , for which the name Prauserella isguenensis sp. nov. is proposed. The type strain is H225(T) ( =DSM 46664(T) = CECT 8577(T)).

  2. Phytohabitans suffuscus gen. nov., sp. nov., an actinomycete of the family Micromonosporaceae isolated from plant roots.

    PubMed

    Inahashi, Yuki; Matsumoto, Atsuko; Danbara, Hirofumi; Omura, Satoshi; Takahashi, Yōko

    2010-11-01

    An actinomycete strain, K07-0523(T), was isolated from the roots of an orchid collected in Okinawa prefecture, Japan. 16S rRNA gene sequence analysis indicated that the new strain belonged to the family Micromonosporaceae and the similarity values between strain K07-0523(T) and the type species of 24 genera in the family Micromonosporaceae were 93.3-97.7 %. Strain K07-0523(T) contained D-glutamic acid, glycine, D-alanine, meso-diaminopimelic acid, hydroxydiaminopimelic acid and L-lysine in the cell wall. The major menaquinones were MK-9(H(6)), MK-10(H(4)) and MK-10(H(6)). Galactose, glucose, mannose, ribose and xylose were present in the whole-cell sugars. The acyl type of the peptidoglycan was glycolyl. Major fatty acids were anteiso-C(17 : 0), iso-C(17 : 0), iso-C(16 : 0) and iso-C(15 : 0). Phosphatidylethanolamine was detected as the major phospholipid and corresponded to phospholipid type II. The G+C content of the genomic DNA was 73 mol%. On the basis of phylogenetic and chemotaxonomic data, the new strain represents a member of a new genus and novel species, namely Phytohabitans suffuscus gen. nov., sp. nov., in the family Micromonosporaceae. The type strain of the type species is K07-0523(T) (=DSM 45306(T)=NBRC 105367(T)).

  3. Isolation of endophytic diazotroph Pantoea agglomerans and nondiazotroph Enterobacter asburiae from sweetpotato stem in Japan.

    PubMed

    Asis, C A; Adachi, K

    2004-01-01

    To isolate and identify diazotrophic endophytes in the stem of Japanese sweetpotato cv. Koganesengan. Surface-sterilized and thinly sliced (1-2 mm) sweetpotato stem samples were incubated in test tubes with semi-solid modified Rennie (MR) medium. The test tubes were assayed for acetylene reduction activity (ARA) 5 days after incubation at 30 degrees C. Twelve isolates were obtained from MR plates inoculated with a loop of semi-solid MR medium from ARA+ tubes. However, ARA test showed that only nine isolates were diazotrophic and three were nondiazotrophic strains. Using the API 20E diagnostic kit, four diazotrophic isolates were identified as strains of Pantoea spp. and five isolates as Klebsiella spp. The nondiazotrophic bacteria were strains of Enterobacter spp. A diazotrophic isolate Pantoea sp. MY1 and nondiazotrophic isolate Enterobacter sp. MY2 were identified to the species level by full sequence analysis of 16S rRNA gene. The results showed that MY1 had 99.2% similarity to Pantoea agglomerans ATCC 27155 and MY2 had 99.5% similarity to Enterobacter asburiae ATCC 35953. The stem of sweetpotato cv. Koganesengan was colonized by diazotrophic endophyte P. agglomerans and nondiazotrophic endophyte E. asburiae. This study is an essential step toward understanding the ecology and interaction between endophytic bacteria and sweetpotato.

  4. Streptomyces canalis sp. nov., an actinomycete isolated from an alkali-removing canal.

    PubMed

    Xie, Yu-Xuan; Han, Xiao-Xue; Luo, Xiao-Xia; Xia, Zhan-Feng; Wan, Chuan-Xing; Zhang, Li-Li

    2016-08-01

    A novel actinomycete strain, designated TRM 46794-61T, was isolated from an alkali-removing canal in 14th Farms of Xinjiang Production and Construction Corps, north-west China. The isolate contained ll-diaminopimelic acid as the diagnostic diamino acid. The whole-cell sugar patterns of the isolate contained ribose, mannose and glucose. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannoside and two unidentified phospholipids. The predominant menaquinones were MK-9(H2), MK-9(H4), MK-9(H6) and MK-9(H8). The major fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. The G+C content of the DNA was 70.4 mol%. Phylogenetic analysis showed that strain TRM 46794-61T had a 16S rRNA gene sequence similarity of 97.6 % with the most closely related species with a validly published name, Streptomyces aidingensis TRM 46012T, and it could be distinguished from all species in the genus Streptomyces based on data from this polyphasic taxonomic study. However, DNA-DNA hybridization studies between strain TRM 46794-61T and S.aidingensis TRM 46012T showed only 45.4 % relatedness. On the basis of these data, strain TRM 46794-61T should be designated as a representative of a novel species of the genus Streptomyces, for which the name Streptomyces canalis sp. nov. is proposed. The type strain is TRM 46794-61T (=CCTCC AA 2015006T=KCTC 39568T).

  5. Saccharopolyspora griseoalba sp. nov., a novel actinomycete isolated from the Dead Sea.

    PubMed

    Jiang, Yingying; Wei, Xiaomin; Chen, Xiu; Jiang, Yi; Xue, Quanhong; Lai, Hangxian; Jiang, Chenglin

    2016-12-01

    A novel halotolerant actinomycete, designated strain AFM 10238(T), was isolated from a sediment sample collected from the Dead Sea of Israel. The isolate grew at 15-45 °C, pH 6-12 and with 0-15 % (w/v) NaCl. Strain AFM 10238(T) contains meso-diaminopimelic acid as cell wall diamino acid, and galactose and arabinose as the whole cell sugars. The major polar lipids are phosphatidylcholine, phosphatidylglycerol, and diphosphatidylglycerol. Major fatty acids are iso-C16:0, iso-C17:0, iso-C15:0, anteiso-C17:0 and C17:1 ω8c. MK-9(H4) is the predominant menaquinone and the DNA G + C content is 72.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM10238(T) belongs to the genus Saccharopolyspora. The 16S rRNA gene sequence similarity between strain AFM 10238(T) and its close neighbours, Saccharopolyspora halophila YIM 90500(T) , Saccharopolyspora spinosa DSM 44228(T), Saccharopolyspora dendranthemae KLBMP 1305(T) and Saccharopolyspora cebuensis DSM 45019(T) were 98.2, 97.2, 97.1 and 97.0 %, respectively. Sequence similarities to other type strains of this genus were below 97 %. DNA-DNA relatedness data, together with phenotypic and chemotaxonomic differences, clearly distinguished the isolate from its close neighbours. On the basis of the data from this polyphasic analysis, a novel species Saccharopolyspora griseoalba sp. nov. is proposed. The type strain is AFM 10238(T) (= DSM 46,663 = CGMCC 4.7124).

  6. Amycolatopsis flava sp. nov., a halophilic actinomycete isolated from Dead Sea.

    PubMed

    Wei, Xiaomin; Jiang, Yingying; Chen, Xiu; Jiang, Yi; Lai, Hangxian

    2015-10-01

    A novel halophilic, filamentous actinomycete, designated strain AFM 10111(T), was isolated from a sediment sample collected from the Dead Sea of Israel and its taxonomic position was established by using a polyphasic taxonomic approach. The isolate grew at 20-35 °C, pH 5-12 and with 1-30 % NaCl. The substrate mycelium is white or yellow, well developed, branched and fragments into squarish, rod-like elements. The isolate contained meso-diaminopimelic acid as cell-wall diamino acid, and arabinose and galactose as whole-cell sugars. The major menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine phosphatidylmethylethanolamine and one unidentified phospholipid. Major fatty acids were iso-C16:0, iso-C16:1 H, C17:1 ω6c. The DNA G + C content was 67.7 mol %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM 10111(T) belongs to the genus Amycolatopsis, and formed a distinct clade with Amycolatopsis marina CGMCC 4.3568(T) and Amycolatopsis palatopharyngis CGMCC 4.1729(T), with the sequence similarity 98.4 and 98.6 %. The level of DNA-DNA relatedness between the strain AFM 10111(T) and A. marina CGMCC 4.3568(T) and A. palatopharyngis CGMCC 4.1729(T) were 46.9 ± 3.08 and 49.4 ± 1.25 %. The combined genotypic and phenotypic data indicate that strain AFM 10111(T) represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis flava sp. nov. is proposed. The type strain is AFM 10111(T) (= DSM 46658(T) = CGMCC 4.7123(T)).

  7. Saccharothrix lopnurensis sp. nov., a filamentous actinomycete isolated from sediment of Lop Nur.

    PubMed

    Li, Yu-Qian; Liu, Lan; Cheng, Cong; Shi, Xiao-Han; Lu, Chun-Yan; Dong, Zhou-Yan; Salam, Nimaichand; An, Deng-Di; Li, Wen-Jun

    2015-10-01

    A novel actinomycete strain, designated YIM LPA2h(T), was isolated from a sediment sample collected from Lop Nur, Xinjiang Uygur Autonomous Region, North-West China. The taxonomic position of strain YIM LPA2h(T) was investigated by a polyphasic approach. The morphological and chemotaxonomic properties of the isolate were in accordance with the members of the genus Saccharothrix. The 16S rRNA gene sequence of the strain showed highest similarities to Saccharothrix yanglingensis (98.6 %), Saccharothrix longispora (98.4 %) and Saccharothrix hoggarensis (98.3 %). However, the DNA-DNA hybridization values between the new isolate YIM LPA2h(T) and S. yanglingensis, S. longispora and S. hoggarensis were significantly below 70 %. Strain YIM LPA2h(T) was found to contain meso-diaminopimelic acid as diagnostic diamino acid. The major sugars in whole-cell hydrolysates were rhamnose, galactose, mannose, glucose and fructose. The major polar lipids were identified as phosphatidylethanolamine, phosphatidylhydroxylethanolamine, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. The predominant respiratory menaquinones were MK-9 (H4) and MK-10 (H4). The major fatty acids were C17:1 ω8c (15 %), iso-C15:0 (12 %), anteiso-C15:0 (12 %) and summed feature 3 (C16:1 ω6c/C16:1 ω7c, 10 %). The genomic DNA G+C content of strain YIM LPA2h(T) was determined to be 75 mol %. The genotypic and phenotypic results suggest that strain YIM LPA2h(T) represents a novel species of the genus Saccharothrix, for which the name Saccharothrix lopnurensis sp. nov. is proposed. The type strain is YIM LPA2h(T) (=CGMCC 4.7246(T)=KCTC 39545(T)).

  8. Actinopolyspora saharensis sp. nov., a novel halophilic actinomycete isolated from a Saharan soil of Algeria.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-04-01

    A novel halophilic actinomycete, strain H32(T), was isolated from a Saharan soil sample collected in El-Oued province, south Algeria. The isolate was characterized by means of polyphasic taxonomy. Optimal growth was determined to occur at 28-32 °C, pH 6.0-7.0 and in the presence of 15-25 % (w/v) NaCl. The strain was observed to produce abundant aerial mycelium, which formed long chains of rod-shaped spores at maturity, and fragmented substrate mycelium. The cell wall was determined to contain meso-diaminopimelic acid and the characteristic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-10(H4) and MK-9(H4). The predominant cellular fatty acids were determined to be anteiso C17:0, iso-C15:0 and iso-C16:0. The diagnostic phospholipid detected was phosphatidylcholine. Phylogenetic analyses based on the 16S rRNA gene sequence showed that this strain formed a distinct phyletic line within the radiation of the genus Actinopolyspora. The 16S rRNA gene sequence similarity indicated that strain H32(T) was most closely related to 'Actinopolyspora algeriensis' DSM 45476(T) (98.8 %) and Actinopolyspora halophila DSM 43834(T) (98.5 %). Furthermore, the result of DNA-DNA hybridization between strain H32(T) and the type strains 'A. algeriensis' DSM 45476(T), A. halophila DSM 43834(T) and Actinopolyspora mortivallis DSM 44261(T) demonstrated that this isolate represents a different genomic species in the genus Actinopolyspora. Moreover, the physiological and biochemical data allowed the differentiation of strain H32(T) from its closest phylogenetic neighbours. Therefore, it is proposed that strain H32(T) represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora saharensis sp. nov. is proposed. The type strain is H32(T) (=DSM 45459(T)=CCUG 62966(T)).

  9. Cytotoxic and antibacterial activities of endophytic fungi isolated from plants at the National Park, Pahang, Malaysia

    PubMed Central

    2009-01-01

    Background Endophytes, microorganisms which reside in plant tissues, have potential in producing novel metabolites for exploitation in medicine. Cytotoxic and antibacterial activities of a total of 300 endophytic fungi were investigated. Methods Endophytic fungi were isolated from various parts of 43 plants from the National Park Pahang, Malaysia. Extracts from solid state culture were tested for cytotoxicity against a number of cancer cell lines using the MTT assay. Antibacterial activity was determined using the disc diffusion method. Results A total of 300 endophytes were isolated from various parts of plants from the National Park, Pahang. 3.3% of extracts showed potent (IC50 < 0.01 μg/ml) cytotoxic activity against the murine leukemic P388 cell line and 1.7% against a human chronic myeloid leukemic cell line K562. Sporothrix sp. (KK29FL1) isolated from Costus speciosus showed strong cytotoxicity against colorectal carcinoma (HCT116) and human breast adenocarcinoma (MCF7) cell lines with IC50 values of 0.05 μg/ml and 0.02 μg/ml, respectively. Antibacterial activity was demonstrated for 8% of the extracts. Conclusion Results indicate the potential for production of bioactive agents from endophytes of the tropical rainforest flora. PMID:19930582

  10. Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity

    PubMed Central

    Astuti, Puji; Sudarsono, Sudarsono; Nisak, Khoirun; Nugroho, Giri Wisnu

    2014-01-01

    Purpose: Coleus amboinicus is a medicinal plant traditionally used to treat various diseases such as throat infection, cough and fever, diarrhea, nasal congestion and digestive problems. The plant was explored for endophytic fungi producing antimicrobial agents. Methods: Screening for endophytic fungi producing antimicrobial agents was conducted using agar plug method and antimicrobial activity of promising ethyl acetate extracts was determined by disc diffusion assay. Thin layer chromatography (TLC) - bioautography was performed to localize the bioactive components within the extract. TLC visualization detection reagents were used to preliminary analyze phytochemical groups of the bioactive compounds. Results: Three endophytic fungi were obtained, two of them showed promising potential. Agar diffusion method showed that endophytic fungi CAL-2 exhibited antimicrobial activity against P. aeruginosa, B. subtilis, S. aureus and S. thypi, whilst CAS-1 inhibited the growth of B. subtilis. TLC bioautography of ethyl acetate extract of CAL-2 revealed at least three bands exhibited antimicrobial activity and at least two bands showed inhibition of B. subtilis growth. Preliminary analysis of the crude extracts suggests that bioactive compounds within CAL-2 extract are terpenoids, phenolics and phenyl propanoid compounds whilst the antimicrobial agents within CAS-1 extract are terpenoids, propylpropanoids, alkaloids or heterocyclic nitrogen compounds. Conclusion: These data suggest the potential of endophytic fungi of C. amboinicus as source for antimicrobial agents. PMID:25671195

  11. Gymnemagenin-producing endophytic fungus isolated from a medicinal plant Gymnema sylvestre R.Br.

    PubMed

    Parthasarathy, Ramalingam; Sathiyabama, Muthukrishnan

    2014-03-01

    Gymnema sylvestre is a plant containing the triterpenoid gymnemagenin, which is used in the pharmaceutical industry as an antidiabetic agent. The objective of this study was to determine whether endophytic fungi, isolated from G. sylvestre, produce gymnemagenin. We isolated an endophytic fungal strain from the leaves of G. sylvestre which produces gymnemagenin in the medium. The fungus was identified as Penicillium oxalicum based on morphological and molecular methods. The strain had a component with the same TLC Rf value and HPLC retention time as authentic gymnemagenin. The presence of gymnemagenin was further confirmed by FTIR, UV, and (1)H NMR analyses.

  12. Molecular analysis of endophytic bacteria from the genus Bacillus isolated from tropical maize (Zea mays L.).

    PubMed

    Figueiredo, José Edson Fontes; Gomes, Eliane Aparecida; Guimarães, Claudia Teixeira; de Paula Lana, Ubiraci Gomes; Teixeira, Marta Aparecida; Lima, Guilherme Vitor Corrêa; Bressan, Wellington

    2009-07-01

    Endophytic bacteria play an important role in agriculture by improving plant performance and adaptation against biotic and abiotic stresses. In the present study molecular methods were used for identifying Bacillus endophytic bacteria isolated from Brazilian sweet corn. SDS-PAGE of whole-cell protein extract of forty-two isolates revealed a high number of scrutinable bands. Twenty-four isolates were identified in nine different groups of duplicated bacteria and eighteen were identified as unique. Some high-accumulated polipeptides with variable length were observed in almost isolates. Partial sequencing of 16S ribosomal gene revealed that all isolates are Bacillus sp. and among thirteen isolates with similar protein profiles, two were different strains. Among the forty-two isolates identified by rDNA sequencing, Bacillus subitilis and B. pumilus were the most frequenty species (15 and 12 isolates, respectively) followed by B. licheniformes (7 isolates), B. cereus (5 isolates) and B. amiloliquefascens (3 isolates). According to present results, SDS-PAGE technique could be used as a fast and cheap first tool for identifying inter-specific variation in maize endophytic bacterial collections while rDNA sequencing could be applied for analyzing intra-specific variation among isolates with similar protein profile as well as for taxonomic studies.

  13. Molecular analysis of endophytic bacteria from the genus Bacillus isolated from tropical maize (Zea mays L.)

    PubMed Central

    Figueiredo, José Edson Fontes; Gomes, Eliane Aparecida; Guimarães, Claudia Teixeira; de Paula Lana, Ubiraci Gomes; Teixeira, Marta Aparecida; Lima, Guilherme Vitor Corrêa; Bressan, Wellington

    2009-01-01

    Endophytic bacteria play an important role in agriculture by improving plant performance and adaptation against biotic and abiotic stresses. In the present study molecular methods were used for identifying Bacillus endophytic bacteria isolated from Brazilian sweet corn. SDS-PAGE of whole-cell protein extract of forty-two isolates revealed a high number of scrutinable bands. Twenty-four isolates were identified in nine different groups of duplicated bacteria and eighteen were identified as unique. Some high-accumulated polipeptides with variable length were observed in almost isolates. Partial sequencing of 16S ribosomal gene revealed that all isolates are Bacillus sp. and among thirteen isolates with similar protein profiles, two were different strains. Among the forty-two isolates identified by rDNA sequencing, Bacillus subitilis and B. pumilus were the most frequenty species (15 and 12 isolates, respectively) followed by B. licheniformes (7 isolates), B. cereus (5 isolates) and B. amiloliquefascens (3 isolates). According to present results, SDS-PAGE technique could be used as a fast and cheap first tool for identifying inter-specific variation in maize endophytic bacterial collections while rDNA sequencing could be applied for analyzing intra-specific variation among isolates with similar protein profile as well as for taxonomic studies. PMID:24031395

  14. Detoxification of Atrazine by Endophytic Streptomyces sp. Isolated from Sugarcane and Detection of Nontoxic Metabolite.

    PubMed

    Mesquini, Josiane A; Sawaya, Alexandra C H F; López, Begonã G C; Oliveira, Valéria M; Miyasaka, Natalia R S

    2015-12-01

    Atrazine is still one of the most used agricultural pesticides worldwide and it has been recognized as a major contaminant of surface and ground water. The aims of this research were to isolate an endophytic microorganism from leaves of sugarcane, evaluate its ability to degrade atrazine, and investigate the formation of metabolites. By sequencing of the 16S rRNA gene, the endophytic isolate atz2 was identified as Streptomyces sp. The reduction in atrazine concentration by Streptomyces sp. atz2 was 98 % and UHPLC-MS/MS analyses showed the appearance of an unknown metabolite observed as m/z 311. Ecotoxicity tests with an aquatic organism, Daphnia similis, confirmed that this metabolite was nontoxic. This mechanism of detoxification of atrazine is different from the ones of other free-living microorganisms that inhabit the soil or rhizosphere. The results show new aspects of atrazine detoxification, highlighting a new role of endophytic bacteria in plants.

  15. Nocardiopsis akesuensis sp. nov., an actinomycete isolated from a salt water beach.

    PubMed

    Gao, Guang-Bin; Luo, Xiao-Xia; Xia, Zhan-Feng; Zhang, Yao; Wan, Chuan-Xing; Zhang, Li-Li

    2016-12-01

    The taxonomic position of a novel actinomycete, strain TRM 46250T, isolated from the sediment of a salt water beach at Baicheng, Xinjiang, China, was determined by a polyphasic approach. Strain TRM 46250T grew optimally in the presence of 2 % (w/v) NaCl and an optimum temperature range for growth of 28-37 °C. The whole-cell sugars of strain TRM 46250T were ribose, xylose, mannose and galactose. The diagnostic diamino acid was meso-diaminopimelic acid. The polar lipids were phosphatidylinositol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylmethyl ethanolamine and six unidentified phospholipids. The predominant menaquinones were MK-10, MK-10(H6) and MK-10(H8). The major fatty acids were 10-methyl C18 : 0, iso-C16 : 0, C16 : 0, iso-G C16 : 1 and C18 : 1ω9c. Based on morphological and chemotaxonomic characteristics the isolate was determined to belong to the genus Nocardiopsis. The phylogenetic tree based on its nearly complete 16S rRNA gene sequence (1493 nt) with those of representative strains showed that the strain consistently falls into a distinct phyletic line together with Nocardiopsis gilva YIM 90087T (97.68 % similarity) and a subclade consisting of Nocardiopsis composta KS9T (97.52 %), Nocardiopsis rosea YIM 90094T (97.44 %) and Nocardiopsis rhodophaea YIM 90096T (97.16 %). However, DNA-DNA hybridization studies between strain TRM 46250T and N. gilva YIM 90087T showed only 36.94 % relatedness. On the basis of these data, strain TRM 46250T should be designated as a representative of a novel species of the genus Nocardiopsis, for which the name Nocardiopsis akesuensis sp. nov. is proposed. The type strain is TRM 46250T (=CCTCC AA 2015027T=KCTC 39725T).

  16. Streptomyces euryhalinus sp. nov., a new actinomycete isolated from a mangrove forest.

    PubMed

    Biswas, Kaushik; Choudhury, Jayanta D; Mahansaria, Riddhi; Saha, Malay; Mukherjee, Joydeep

    2017-06-01

    A Gram-positive, aerobic, non-motile actinomycete (strain MS 3/20(T)) was isolated from the sediment of the Sundarbans mangrove forest in India. On International Streptomyces Project (ISP) medium 2, the isolate produced yellowish brown to red aerial hyphae that carried spiny-surfaced spores in a retinaculum-apertum arrangement. Whole-cell hydrolysate of the strain contained LL-diaminopimelic acid and galactose. Predominant menaquinones were MK-9(H8) and MK-9(H6). Diagnostic polar lipids were glycolipid, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, unidentified phospholipid and unidentified amino lipid. The major fatty acids were anteiso-C15:0 (17.53%), iso-C16:0 (23.89%) and anteiso-C17:0 (10.29%). The strain showed 100% 16S ribosomal RNA (rRNA) gene sequence similarity with Streptomyces variabilis NBRC 12825(T), Streptomyces erythrogriseus LMG 19406(T), Streptomyces griseoincarnatus LMG 19316(T) and Streptomyces labedae NBRC 15864(T). However, strain MS 3/20(T) could be distinguished from these and seven other closely related species based on low levels of DNA-DNA relatedness (27.2-53.8%), supported by the unique banding pattern obtained from random amplified polymorphic DNA-PCR amplification and the distinctive matrix-assisted laser desorption/ionization-time-of-flight/mass spectrometry (MALDI-TOF/MS) profile of whole-cell proteins acquired for strain MS 3/20(T) in comparison with its phylogenetic relatives. Disparate morphological, physiological and chemotaxonomic features, principally growth in NaCl, further corroborated the distinction of strain MS 3/20(T) from other phylogenetic relatives. Strain MS 3/20(T) is therefore suggested to be a novel species of the genus Streptomyces, for which the name Streptomyces euryhalinus sp. nov. is proposed. The type strain is MS 3/20(T) (=CICC 11032(T)=DSM 103378(T)).

  17. Nonomuraea flavida sp. nov., a novel species of soil actinomycete isolated from Aconitum napellus rhizosphere.

    PubMed

    Chen, Shaofeng; Shi, Jindi; Li, Dan; Wu, Yingying; Huang, Yaojian

    2015-11-01

    A novel actinomycete strain, YN-5-1T, isolated from the rhizosphere soil of a medicinal plant, Aconitum napellus, was characterized by a polyphasic approach to determine its taxonomic position. The strain showed highest 16S rRNA gene sequence similarities of 97.3, 97.2 and 97.1 % to Nonomuraea turkmeniaca DSM 43926T, Nonomuraea ferruginea DSM 43553T and Nonomuraea candida DSM 45086T, respectively. A wide range of genotypic and phenotypic characteristics, as well as levels of DNA-DNA relatedness between strain YN-5-1T and N. turkmeniaca DSM 43926T (57.46 %), N. ferruginea DSM 43553T (53.50 %) and N. candida DSM 45086T (48.80 %), distinguished the novel isolate from its closest phylogenetic neighbours. The morphological characteristics of strain YN-5-1T were typical of the genus Nonomuraea. Chemotaxonomic characteristics, such as diagnostic diamino acid of the peptidoglycan, whole-cell sugars, phospholipid type, major menaquinone and major fatty acids, further supported the assignment of strain YN-5-1T to the genus Nonomuraea. The G+C content of the genomic DNA was 72.1 mol%. Based on the above data, strain YN-5-1T is considered to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea flavida sp. nov. is proposed. The type strain is YN-5-1T ( = CCTCC AB 2012909T = KCTC 29143T).

  18. Phylogenetic diversity of endophytic leaf fungus isolates from the medicinal tree Trichilia elegans (Meliaceae).

    PubMed

    Rhoden, S A; Garcia, A; Rubin Filho, C J; Azevedo, J L; Pamphile, J A

    2012-08-16

    Various types of organisms, mainly fungi and bacteria, live within vegetal organs and tissues, without causing damage to the plant. These microorganisms, which are called endophytes, can be useful for biological control and plant growth promotion; bioactive compounds from these organisms may have medical and pharmaceutical applications. Trichilia elegans (Meliaceae) is a native tree that grows abundantly in several regions of Brazil. Preparations using the leaves, seeds, bark, and roots of many species of the Meliaceae family have been widely used in traditional medicine, and some members of the Trichilia genus are used in Brazilian popular medicine. We assessed the diversity of endophytic fungi from two wild specimens of T. elegans, collected from a forest remnant, by sequencing ITS1-5.8S-ITS2 of rDNA of the isolates. The fungi were isolated and purified; 97 endophytic fungi were found; they were separated into 17 morpho-groups. Of the 97 endophytic fungi, four genera (Phomopsis, Diaporthe, Dothideomycete, and Cordyceps) with 11 morpho-groups were identified. Phomopsis was the most frequent genus among the identified endophytes. Phylogenetic analysis showed two major clades: Sordariomycetes, which includes three genera, Phomopsis, Diaporthe, and Cordyceps, and the clade Dothideomycetes, which was represented by the order Pleosporales.

  19. Antifungal and antiproliferative activities of endophytic fungi isolated from the leaves of Markhamia tomentosa.

    PubMed

    Ibrahim, Mutiat; Kaushik, Nutan; Sowemimo, Abimbola; Chhipa, Hemraj; Koekemoer, Trevor; van de Venter, Maryna; Odukoya, Olukemi A

    2017-12-01

    Plants harbor endophytes with potential bioactivity. Markhamia tomentosa (Benth) K. Schum ex. Engl. (Bignoniaceae) is reported to possess antioxidant, anti-inflammatory and anticancer activities. The antifungal and antiproliferative properties of endophytic fungi extracts and fractions from M. tomentosa were evaluated. Endophytic fungi were isolated from the leaves of M. tomentosa and identified by ITS-rDNA sequence analysis. The antagonistic effect of the fungal strains was investigated against pathogenic fungi viz, Fusarium oxysporum, Sclerotinia sclerotiorium, Rhizoctonia solani, and Botrytis cinerea using the dual culture assay for 5-7 days. Antiproliferative effect of the fungal extracts and fractions (3.91-250 μg/mL) on HeLa cancer cell line was tested and IC50 was calculated. Poisoning food assay and antifeedant activity against the pathogenic fungi and Spodoptera litura larvae, for 7 days and 2 h, respectively, was also tested at concentrations of 250, 500 and 1000 μg/mL. Fungal endophytes Trichoderma longibrachiatum and Syncephalastrum racemosum were isolated from the leaves of M. tomentosa. Isolated endophytic fungal strains and solvent extracts showed MIC value of 1000 μg/mL against tested pathogenic fungi in the dual culture and poisoning food assays. Methanol fraction of S. racemosum isolate showed the most effective antiproliferative activity with IC50 of 43.56 μg/mL. Minimal feeding deterrent activity against S. litura larvae was also observed. These findings showed that the leaves of Markhamia tomentosa harbor strains of endophytic fungi with promising health benefits, and suggest their antifungal and antiproliferative effects against pathogenic fungi and HeLa cancer cell line.

  20. [Isolation of endophytic fungi from medicinal plant Brucea javanica and their microbial inhibition activity].

    PubMed

    Liang, Zi-Ning; Zhu, Hua; Lai, Kai-Ping; Chen, Long

    2014-04-01

    To isolate and identify endophytic fungi from Brucea javanica, and to detect the antimicrobial activity of these strains. Endophytic fungi were isolated by tissue inoculation culture and identified by conventional morphological characteristic method. Seven kinds of pathogenic fungi and three kinds of bacteria were used as targeting microbes to test microbial inhibition activities by agar plate antagonistic action and modified agar gel diffusion methods, respectively. A total of 83 endophytic fungi strains were isolated from the root, stem, leaf and fruit of Brucea javanica. 34 strains were obtained from the stem, 32 strains were obtained from the leaf, 15 strains were isolated from the root and 2 strains came from the fruit. These 73 strains which had been identified attribute to 5 orders, 6 families and 12 genera. For the isolated strains, 14 strains had antifungal activities against at least one pathogenic fungi, 9 strains showed antibacterial activities against one or more bacteria. Especially, the strain YJ-17 which belonged to Phomopsis genus showed the best inhibitory effect on the targeting microbes. The endophytic fungi from Brucea javanica show diversity and microbial inhibition activity, and are worthy for further study on plant disease controlling.

  1. Plant growth promotion induced by phosphate solubilizing endophytic Pseudomonas isolates

    PubMed Central

    Oteino, Nicholas; Lally, Richard D.; Kiwanuka, Samuel; Lloyd, Andrew; Ryan, David; Germaine, Kieran J.; Dowling, David N.

    2015-01-01

    The use of plant growth promoting bacterial inoculants as live microbial biofertilizers provides a promising alternative to chemical fertilizers and pesticides. Inorganic phosphate solubilization is one of the major mechanisms of plant growth promotion by plant associated bacteria. This involves bacteria releasing organic acids into the soil which solubilize the phosphate complexes converting them into ortho-phosphate which is available for plant up-take and utilization. The study presented here describes the ability of endophytic bacteria to produce gluconic acid (GA), solubilize insoluble phosphate, and stimulate the growth of Pisum sativum L. plants. This study also describes the genetic systems within three of these endophyte strains thought to be responsible for their effective phosphate solubilizing abilities. The results showed that many of the endophytic strains produced GA (14–169 mM) and have moderate to high phosphate solubilization capacities (~400–1300 mg L−1). When inoculated into P. sativum L. plants grown in soil under soluble phosphate limiting conditions, the endophytes that produced medium-high levels of GA displayed beneficial plant growth promotion effects. PMID:26257721

  2. Whole-genome sequence of Enterobacter sp. strain SST3, an endophyte isolated from Jamaican sugarcane (Saccharum sp.) stalk tissue.

    PubMed

    Gan, Han Ming; McGroty, Sean E; Chew, Teong Han; Chan, Kok Gan; Buckley, Larry J; Savka, Michael A; Hudson, André O

    2012-11-01

    Enterobacter sp. strain SST3 is an endophytic bacterium isolated from Saccharum spp. Here we present its annotated draft genome that may shed light on its role as a bacterial endophyte of sugarcane. To our knowledge, this is the first genome announcement of a sugarcane-associated bacterium from the genus Enterobacter.

  3. Actinoplanes rhizophilus sp. nov., an actinomycete isolated from the rhizosphere of Sansevieria trifasciata Prain.

    PubMed

    He, Hairong; Xing, Jia; Liu, Chongxi; Li, Chuang; Ma, Zhaoxu; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing

    2015-12-01

    A novel actinomycete, designated strain NEAU-A-2T, was isolated from the rhizosphere soil of Sansevieria trifasciata Prain collected from Heilongjiang province, north-east China. The taxonomic status of this organism was established using a polyphasic approach. The isolate formed irregular sporangia containing motile spores on the substrate mycelium. The whole-cell sugars were xylose and galactose. The predominant menaquinones were MK-9(H10), MK-9(H2), MK-10(H2) and MK-10(H4). The major fatty acids were iso-C15 : 0, iso-C16 : 0 and anteiso-C15 : 0. The polar lipids were diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, three unidentified phospholipids and an unidentified glycolipid. 16S rRNA gene sequence similarity studies showed that strain NEAU-A-2T belongs to the genus Actinoplanes with the highest sequence similarities to Actinoplanes globisporus NBRC 13912T (97.7 % 16S rRNA gene sequence similarity), Actinoplanes ferrugineus IMSNU 22125T (97.5 %), Actinoplanes toevensis MN07-A0368T (97.2 %) and Actinoplanes rishiriensis NBRC 108556T (97.2 %); similarities to type strains of other species of this genus were < 97 %. Two tree-making algorithms showed that strain NEAU-A-2T formed a distinct clade with A. globisporus NBRC 13912T and A. rishiriensis NBRC 108556T. However, low DNA-DNA relatedness values allowed the isolate to be differentiated from the above-mentioned two species of the genus Actinoplanes. Moreover, strain NEAU-A-2T could also be distinguished from the most closely related species by morphological and physiological characteristics. Therefore, in conclusion, isolate NEAU-A-2T represents a novel species of the genus Actinoplanes, for which the name Actinoplanes rhizophilus sp. nov. is proposed. The type strain is NEAU-A-2T ( = CGMCC 4.7133T = DSM 46672T).

  4. Actinopolyspora biskrensis sp. nov., a novel halophilic actinomycete isolated from Northern Sahara.

    PubMed

    Saker, Rafika; Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2015-03-01

    A novel halophilic, filamentous actinomycete, designated H254(T), was isolated from a Saharan soil sample collected from Biskra (Northern Sahara), and subjected to a polyphasic taxonomic characterization. The strain is Gram-positive, aerobic, and halophilic, and the optimum NaCl concentration for growth is 15-20 % (w/v). The cell-wall hydrolysate contained meso-diaminopimelic acid, and the diagnostic whole-cell sugars were arabinose and galactose. The diagnostic phospholipid detected was phosphatidylcholine, and MK-9(H4) was the predominant menaquinone. The major fatty acid profiles were anteiso-C17:0 (32.8 %), C15:0 (28 %), and iso-C17:0 (12.3 %). Comparative analysis of the 16S rRNA gene sequences revealed that the strain H254(T) formed a well-separated sub-branch within the radiation of the genus Actinopolyspora, and the microorganism was most closely related to Actinopolyspora saharensis DSM 45459(T) (99.2 %), Actinopolyspora halophila DSM 43834(T) (99.1 %), and Actinopolyspora algeriensis DSM 45476(T) (99.0 %). Nevertheless, the strain had relatively lower mean values for DNA-DNA relatedness with the above strains (57.2, 68.4, and 45.6 %, respectively). Based on phenotypic features and phylogenetic position, we propose that strain H254(T) represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora biskrensis sp. nov. is proposed. The type strain of A. biskrensis is strain H254(T) (=DSM 46684(T) =CECT 8576(T)).

  5. Nonomuraea indica sp. nov., novel actinomycetes isolated from lime-stone open pit mine, India.

    PubMed

    Quadri, Syed Raziuddin; Tian, Xin-Peng; Zhang, Jing; Li, Jie; Nie, Guo-Xing; Tang, Shu-Kun; Al Ruwaili, Jamal; Agsar, Dayanand; Li, Wen-Jun; Dastager, Syed G

    2015-08-01

    A Gram-positive, aerobic, nonmotile actinomycete strain designated DRQ-2(T) was isolated from the soil sample collected from lime-stone open pit mine from the Gulbarga region, Karnataka province, India. Strain DRQ-2(T) was identified as a member of the genus Nonomuraea by a polyphasic approach. Strain DRQ-2(T) could be differentiated from other members of the genus Nonomuraea on the basis of physiology and 16S rRNA gene sequence analysis. The 16S rRNA gene sequence similarity of strain DRQ-2(T) showed highest sequence similarity to Nonomuraea muscovyensis DSM 45913(T) (99.1%), N. salmonea DSM 43678(T) (98.2%) and N. maheshkhaliensis JCM 13929(T) with 98.0%, respectively. Chemotaxonomic properties showing predominant menaquinones of MK-9 (H4), MK-9(H2) and MK-9(H6), major polar lipids comprised diphosphatidylglycerol, phosphatidylmono methyl ethanolamine (PME), phosphatidylethanolamine (PE), hydroxy-PME (OH-PME), hydroxy PE (OH-PEE), phosphatidylglycerol (PG), ninhydrin-positive phosphoglycolipid and unknown phospholipid, fatty acids with major amounts of i-C16:0, ai-C15:0 and ai-C17:0 supported allocation of the strain to the genus Nonomuraea. Results of DNA-DNA hybridization and physiological tests allowed genotypic and phenotypic differentiation of strain DRQ-2(T) from closely related species. The genomic DNA G+C content of the organism was 72.5 mol%. On the basis of phenotypic, chemotypic and molecular characteristics, strain DRQ-2(T) represents a novel species of the genus Nonomuraea, for which the name N. indica sp. nov. is proposed, with type strain DRQ-2(T) (=NCIM 5480(T)= CCTCC AA 209050(T)).

  6. Cryptosporangium eucalypti sp. nov., an actinomycete isolated from Eucalyptus camaldulensis roots.

    PubMed

    Himaman, Winanda; Suksaard, Paweena; Mingma, Ratchanee; Matsumoto, Atsuko; Duangmal, Kannika

    2017-08-01

    A novel actinomycete, designated strain EURKPP3H10T, was isolated from surface-sterilized roots of Eucalyptus camaldulensis Dehnh., collected from Kamphaengphet Silvicultural Research Station, Kamphaengphet province, Thailand. The taxonomic position of strain EURKPP3H10T was studied using a polyphasic approach. Phylogenetic evaluation based on 16S rRNA gene sequence analysis showed that strain EURKPP3H10T belongs to the genus Cryptosporangium, with the highest sequence similarity to Cryptosporangium cibodasense LIPI11-2-Ac046T (99.2 %). Colonies of strain EURKPP3H10T were orange yellow. Spherical sporangia with motile spores were observed. The strain contained meso-diaminopimelic acid and acofriose, arabinose, galactose, glucose, mannose, xylose and ribose in whole-cell hydrolysates. The predominant menaquinones were MK-9(H8) and MK-9(H6). The major fatty acids were iso-C16 : 0, C17 : 1ω8c, C18 : 1ω9c and C17 : 0. The polar lipids of the strain were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and unknown lipids. The DNA G+C content of the genomic DNA was 71.5 mol%. Based on comparative analysis of physiological, biochemical and chemotaxonomic data, including DNA-DNA hybridization, strain EURKPP3H10T represents a novel species of the genus Cryptosporangium, for which the name Cryptosporangium eucalypti sp. nov. is proposed. The type strain is EURKPP3H10T (=BCC 77605T=NBRC 111482T).

  7. Nocardia halotolerans sp. nov., a halotolerant actinomycete isolated from saline soil.

    PubMed

    Moshtaghi Nikou, Mahdi; Ramezani, Mohaddaseh; Ali Amoozegar, Mohammad; Rasooli, Mehrnoosh; Harirchi, Sharareh; Shahzadeh Fazeli, Seyed Abolhasan; Schumann, Peter; Spröer, Cathrin; Ventosa, Antonio

    2015-09-01

    A novel halotolerant actinomycete, strain Chem15(T), was isolated from soil around Inche-Broun hypersaline wetland; its taxonomic position was determined based on a polyphasic approach. Strain Chem15(T) was strictly aerobic and tolerated NaCl up to 12.5%. The optimum temperature and pH for growth were 28-30 °C and pH 7.0-7.5, respectively. The cell wall of strain Chem15(T) contained meso-diaminopimelic acid as diamino acid and galactose, arabinose and ribose as whole-cell sugars. The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The cellular fatty acids profile consisted of C16 : 0, iso-C18 : 0, C18 : 0 10-methyl and C18 : 1ω9c, and the major respiratory quinone was MK-8(H4cycl). The G+C content of the genomic DNA was 68.0 mol%. The novel strain constituted a distinct phyletic line within the genus Nocardia, based on 16S rRNA gene sequence analysis, and was closely associated with Nocardia sungurluensis DSM 45714(T) and Nocardia alba DSM 44684(T) (98.2 and 98.1% 16S rRNA gene sequence similarity, respectively). However DNA-DNA relatedness and phenotypic data demonstrated that strain Chem15(T) was clearly different from closely related species of the genus Nocardia. It is concluded that the organism should be classified as a representative of a novel species of the genus Nocardia, for which the name Nocardia halotolerans sp. nov. is proposed. The type strain is Chem15(T) ( = IBRC-M 10490(T) = LMG 28544(T)).

  8. Streptomyces formicae sp. nov., a novel actinomycete isolated from the head of Camponotus japonicus Mayr.

    PubMed

    Bai, Lu; Liu, Chongxi; Guo, Lifeng; Piao, Chenyu; Li, Zhilei; Li, Jiansong; Jia, Feiyu; Wang, Xiangjing; Xiang, Wensheng

    2016-02-01

    During a screening for novel and biotechnologically useful actinobacteria in insects, a novel actinomycete with antifungal activity, designated strain 1H-GS9(T), was isolated from the head of a Camponotus japonicus Mayr ant, which were collected from Northeast Agricultural University (Harbin, Heilongjiang, China). Strain 1H-GS9(T) was characterised using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain 1H-GS9(T) belongs to the genus Streptomyces with high sequence similarities to Streptomyces scopuliridis DSM 41917(T) (98.8 %) and Streptomyces mauvecolor JCM 5002(T) (98.6 %). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it forms a monophyletic clade with Streptomyces kurssanovii JCM 4388(T) (98.6 %), Streptomyces xantholiticus JCM 4282(T) (98.6 %) and Streptomyces peucetius JCM 9920(T) (98.5 %). Thus, a combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 1H-GS9(T) and the above-mentioned five strains, which further clarified their relatedness and demonstrated that strain 1H-GS9(T) could be distinguished from these strains. Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces formicae sp. nov. is proposed. The type strain is 1H-GS9(T) (=CGMCC 4.7277(T) = DSM 100524(T)).

  9. Streptomyces camponoticapitis sp. nov., an actinomycete isolated from the head of an ant (Camponotus japonicus Mayr).

    PubMed

    Li, Yao; Ye, Lan; Wang, Xiangjing; Zhao, Junwei; Ma, Zhaoxu; Yan, Kai; Xiang, Wensheng; Liu, Chongxi

    2016-10-01

    A novel single-spore-producing actinomycete, designated strain 2H-TWYE14T, was isolated from the head of an ant (Camponotus japonicus Mayr) and characterized using a polyphasic approach. 16S rRNA gene sequence analysis showed that strain 2H-TWYE14T belongs to the genus Streptomyces, with highest sequence similarity to Streptomyces niveus NRRL 2466T (98.84 %). Analysis based on the gyrB gene also indicated that strain 2H-TWYE14T should be assigned to the genus Streptomyces. The chemotaxonomic properties of strain 2H-TWYE14T were consistent with those of members of the genus Streptomyces. The cell wall contained ll-diaminopimelic acid. The predominant menaquinones were MK-9(H6), MK-9(H8) and MK-9(H4). The phospholipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were iso-C16 : 0 and iso-C15 : 0. DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 2H-TWYE14T and its phylogenetically closely related strain S. niveus JCM 4251T, which further clarified their relatedness and demonstrated that 2H-TWYE14T could be distinguished from S. niveus. Therefore, it is concluded that strain 2H-TWYE14T can be classified as representing a novel species of the genus Streptomyces, for which the name Streptomyces camponoticapitis sp. nov. is proposed. The type strain is 2H-TWYE14T (=DSM 100523T=CGMCC 4.7275T).

  10. Salinactinospora qingdaonensis gen. nov., sp. nov., a halophilic actinomycete isolated from a salt pond.

    PubMed

    Chang, Xianbo; Liu, Wenzheng; Zhang, Xiao-Hua

    2012-04-01

    A novel halophilic, filamentous, actinomycete strain, designated CXB832T, was isolated from a salt pond in Qingdao, China. Optimal growth occurred at 37 °C, pH 7.0-8.0 and 9-12 % (w/v) NaCl. Strain CXB832T formed pale yellow to deep yellow branched substrate mycelium without fragmentation. Abundant white aerial mycelium differentiated into long chains of spores and the spores were rod-shaped with smooth surfaces. Strain CXB832T contained meso-diaminopimelic acid as the diagnostic diamino acid of the cell-wall peptidoglycan, and glucose and xylose as the major whole-cell sugars. The phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phospholipids, glycolipid and unidentified lipids. MK-10(H8), MK-9(H8), MK-10(H2) and MK-10(H6) were the predominant menaquinones. The major fatty acids were i-C16:0 (30.71 %), ai-C17:0 (13.31 %) and C16:0 (11.28 %). The G+C content of the DNA was 60.1 mol%. Comparative analysis of 16S rRNA gene sequences showed that the novel strain was most closely related to genera within the family Nocardiopsaceae, but formed a separate lineage. The highest sequence similarities were to Nocardiopsis arabia DSM 45083T (95.4 %) and Haloactinospora alba DSM 45015T (94.9 %). On the basis of phenotypic, chemotaxonomic and phylogenetic distinctiveness, strain CXB832T represents a new genus and novel species in the family Nocardiopsaceae, for which the name Salinactinospora qingdaonensis gen. nov., sp. nov. is proposed. The type strain of the type species is CXB832T (=DSM 45442T=LMG 25567T).

  11. Nocardiopsis algeriensis sp. nov., an alkalitolerant actinomycete isolated from Saharan soil.

    PubMed

    Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Mathieu, Florence; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-02-01

    An alkalitolerant actinomycete strain, designated B32(T), was isolated from a Saharan soil sample collected from Adrar province (South of Algeria), and then investigated using a polyphasic taxonomic approach. The strain was observed to produce short chains of spores on the dichotomous branched aerial mycelium and formed a fragmented substrate mycelium. The optimum NaCl concentration for growth was found to be 0-5 % (w/v) and the optimum growth temperature and pH were found to be 25-35 °C and 7.0-10.0 °C, respectively. The diagnostic diamino acid in the cell-wall peptidoglycan was identified as meso-diaminopimelic acid. The predominant menaquinones of strain B32(T) were identified as MK-10 (H4) and MK-11 (H4). The major fatty acids were found to be iso-C16:0 and anteiso-C15:0. The diagnostic phospholipids detected were phosphatidylcholine, phosphatidylmethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The chemotaxonomic properties of strain B32(T) are consistent with those shared by members of the genus Nocardiopsis. 16S rRNA gene sequence analysis indicated that strain B32(T) is most closely related to Nocardiopsis alba DSM 43377(T) (98.7 %), Nocardiopsis lucentensis DSM 44048(T) (98.6 %), Nocardiopsis aegyptia DSM 44442(T) (98.6 %), Nocardiopsis sinuspersici HM6(T) (98.6 %) and Nocardiopsis arvandica HM7(T) (98.5 %). However, the DNA-DNA relatedness values between strain B32(T) and the closely related type strains were 17.9, 14.6, 31.1, 27.1 and 14.1 %, respectively. Based on the combined genotypic and phenotypic evidence, it is proposed that strain B32(T) should be classified as representative of a novel species, for which the name Nocardiopsis algeriensis sp. nov. is proposed. The type strain is B32(T) (=DSM 45462(T) = CECT 8712(T)).

  12. Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Algeria.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-09-01

    A novel halophilic actinomycete strain, H23(T), was isolated from a Saharan soil sample collected in Djamâa (Oued Righ region), El-Oued province, South Algeria. Strain H23(T) was identified as a member of the genus Actinopolyspora by a polyphasic approach. Phylogenetic analysis showed that strain H23(T) had 16S rRNA gene sequence similarities ranging from 97.8 % (Actinopolyspora xinjiangensis TRM 40136(T)) to 94.8 % (Actinopolyspora mortivallis DSM 44261(T)). The strain grew optimally at pH 6.0-7.0, 28-32 °C and in the presence of 15-25 % (w/v) NaCl. The substrate mycelium was well developed and fragmented with age. The aerial mycelium produced long, straight or flexuous spore chains with non-motile, smooth-surfaced and rod-shaped spores. Strain H23(T) had MK-10 (H4) and MK-9 (H4) as the predominant menaquinones. The whole micro-organism hydrolysates mainly consisted of meso-diaminopimelic acid, galactose and arabinose. The diagnostic phospholipid detected was phosphatidylcholine. The major cellular fatty acids were anteiso-C17:0 (37.4 %), iso-C17:0 (14.8 %), iso-C15:0 (14.2 %), and iso-C16:0 (13.9 %). The genotypic and phenotypic data show that the strain represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora righensis sp. nov. is proposed, with the type strain H23(T) (=DSM 45501(T) = CCUG 63368(T) = MTCC 11562(T)).

  13. Streptomonospora algeriensis sp. nov., a halophilic actinomycete isolated from soil in Algeria.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Riba, Amar; Zitouni, Abdelghani; Mathieu, Florence; Rohde, Manfred; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2014-08-01

    A halophilic actinomycete strain, designated H27(T), was isolated from a soil sample collected from a hypersaline habitat in Djelfa Province (North-Central Algeria), and then investigated using a polyphasic taxonomic approach. The strain was observed to produce poor aerial mycelium, which formed short chains of oval to cylindrical-shaped spores at maturity, and non fragmented substrate mycelium. The optimum NaCl concentration for growth was found to be 10-15 % (w/v) and the optimum growth temperature and pH were found to be 28-37 °C and 6-7, respectively. The diagnostic diamino acid in the cell-wall peptidoglycan was identified as meso-diaminopimelic acid. The predominant menaquinones of strain H27(T) were identified as MK-11 (H4) and MK-10 (H6). The major fatty acids were found to be iso-C16:0, anteiso-C17:0, 10 methyl C17:0 and 10 methyl C16:0. The diagnostic phospholipids detected were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylinositol. The chemotaxonomic properties of strain H27(T) are consistent with those shared by members of the genus Streptomonospora. 16S rRNA gene sequence analysis indicated that strain H27(T) is most closely related to Streptomonospora alba DSM 44588(T) (98.8 %) and Streptomonospora flavalba DSM 45155(T) (98.7 %) whereas the DNA-DNA relatedness values between strain H27(T) and the two type strains were 17.1 and 57.9 %, respectively. Based on the combined genotypic and phenotypic evidence, it is proposed that strain H27(T) should be classified as representative of a novel species, for which the name Streptomonospora algeriensis sp. nov. is proposed. The type strain is H27(T) (=DSM 45604(T) =CCUG 63369(T) =MTCC 11563(T)).

  14. Saccharopolyspora ghardaiensis sp. nov., an extremely halophilic actinomycete isolated from Algerian Saharan soil.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Sabaou, Nasserdine; Mathieu, Florence; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter

    2014-04-01

    A novel halophilic actinomycete, strain designated H53(T), was isolated from a Saharan soil sample collected from Chaâbet Ntissa, Béni-isguen, Ghardaïa (South of Algeria) and was characterized taxonomically by means of polyphasic approach. Optimal growth was found to occur at 30-35 °C, pH 6-7 and in the presence of 15-25% (w/v) NaCl. The strain was observed to produce abundant aerial mycelium, which formed long chains of rod-shaped spores at maturity, and well developed and fragmented substrate mycelium. The cell wall was determined to contain meso-diaminopimelic acid; the diagnostic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-9(H₄) and MK-9(H₆). The predominant cellular fatty acids were determined to be iso- and anteiso-C17:0, iso-C15:0, and cis9 iso-C17:1. The diagnostic phospholipid detected was phosphatidylcholine. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Saccharopolyspora. Phylogenetic analyses on the basis of the 16S ribosomal RNA (rRNA) gene sequence showed that this strain formed a distinct phyletic line within the radiation of the genus Saccharopolyspora. The 16S rRNA sequence similarities between strain H53(T) and other members of the genus Saccharopolyspora ranged from 92.1 to 94.3%. The DNA G+C content of strain H53(T) was 72.6%. The genotypic and phenotypic data showed that the strain H53(T) represents a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora ghardaiensis sp. nov. is proposed, with the type strain H53(T) (=DSM 45606(T)=CCUG 63370(T)=CECT 8304(T)).

  15. Actinokineospora mzabensis sp. nov., a novel actinomycete isolated from Saharan soil.

    PubMed

    Aouiche, Adel; Bouras, Noureddine; Mokrane, Salim; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-01-01

    A novel actinomycete strain, designated PAL84, was isolated from a Saharan soil sample collected from Béni-Isguen, Ghardaïa (South of Algeria). This strain was studied for its taxonomic position using a polyphasic approach and was identified as a member of the genus Actinokineospora. Phylogenetic analysis showed that strain PAL84 had 16S rRNA gene sequence similarities with members of the genus Actinokineospora ranging from 96.2 % (Actinokineospora inagensis DSM 44258(T)) to 97.8 % (Actinokineospora baliensis NBRC 104211(T)). The strain was observed to produce pinkish-purple aerial mycelium and purplish red substrate mycelium, which fragmented readily into chains of non-motile elements. The optimum growth temperature and pH were found to be 25-30 °C and 5.0-7.0, respectively. The cell-wall hydrolysate of strain PAL84 was found to contain meso-diaminopimelic acid and the diagnostic whole-cell sugars were identified as arabinose and galactose. The predominant menaquinone was identified as MK-9 (H4). The major fatty acids were found to be iso-C16:0, iso-C15:0, iso-C16:1 H and iso-C16:0 2OH. The diagnostic phospholipid detected was phosphatidylethanolamine. The genotypic and phenotypic data show that the strain represents a novel species of the genus Actinokineospora, for which the name Actinokineospora mzabensis sp. nov. is proposed, with the type strain PAL84(T) (=DSM 45961(T) = CECT 8578(T)).

  16. Metabolites from the endophytic fungus Sporormiella minimoides isolated from Hintonia latiflora

    USDA-ARS?s Scientific Manuscript database

    An extract of the solid cultures of Sporormiella minimoides (Sporormiaceae) isolated as an endophytic fungus from Hintonia latiflora (Rubiaceae), yielded three polyketides, 3,6-dimethoxy-8-methyl-1H,6H-benzo[de]isochromene-1,9-dione, 3-hydroxy-1,6,10-trimethoxy-8-methyl-1H,3H-benzo[de]isochromen-9-o...

  17. (+)-Ascosalitoxin and vermelhotin, a calmodulin inhibitor, from an endophytic fungus isolated from Hintonia latiflora

    USDA-ARS?s Scientific Manuscript database

    Chemical investigation of the endophytic fungus 39140-2, isolated from the medicinal plant Hintonia latiflora, yielded the known polyketide vermelhotin (1) and a new salycilic aldehyde derivative, namely 9S,11R-(+)-ascosalitoxin (2). The structure and absolute configuration of the new compound was ...

  18. Acremonium camptosporum isolated as an endophyte of Bursera simaruba from Yucatan Peninsula, Mexico

    USDA-ARS?s Scientific Manuscript database

    This paper draws on morphological and molecular analyses to determinate the systematic position of an interesting endophytic fungus isolated from the leaves of Bursera simaruba, a tree of semideciduous dry tropical forest at El Eden Ecological Reserve. The cultured strain develops the characteristic...

  19. Morphological, Physiological, and Taxonomic Characterization of Actinobacterial Isolates Living as Endophytes of Cacao Pods and Cacao Seeds

    PubMed Central

    Tchinda, Romaric Armel Mouafo; Boudjeko, Thaddée; Simao-Beaunoir, Anne-Marie; Lerat, Sylvain; Tsala, Éric; Monga, Ernest; Beaulieu, Carole

    2016-01-01

    Vascular plants are commonly colonized by endophytic actinobacteria. However, very little is known about the relationship between these microorganisms and cacao fruits. In order to determine the physiological and taxonomic relationships between the members of this community, actinobacteria were isolated from cacao fruits and seeds. Among the 49 isolates recovered, 11 morphologically distinct isolates were selected for further characterization. Sequencing of the 16S rRNA gene allowed the partition of the selected isolates into three phylogenetic clades. Most of the selected endophytic isolates belonged to the Streptomyces violaceusniger clade. Physiological characterization was carried out and a similarity index was used to cluster the isolates. However, clustering based on physiological properties did not match phylogenetic lineages. Isolates were also characterized for traits commonly associated with plant growth-promoting bacteria, including antibiosis and auxin biosynthesis. All isolates exhibited resistance to geldanamycin, whereas only two isolates were shown to produce this antibiotic. Endophytes were inoculated on radish seedlings and most isolates were found to possess plant growth-promoting abilities. These endophytic actinobacteria inhibited the growth of various plant pathogenic fungi and/or bacteria. The present study showed that S. violaceusniger clade members represent a significant part of the actinobacterial community living as endophytes in cacao fruits and seeds. While several members of this clade are known to be geldanamycin producers and efficient biocontrol agents of plant diseases, we herein established the endophytic lifestyle of some of these microorganisms, demonstrating their potential as plant health agents. PMID:26947442

  20. Morphological, Physiological, and Taxonomic Characterization of Actinobacterial Isolates Living as Endophytes of Cacao Pods and Cacao Seeds.

    PubMed

    Tchinda, Romaric Armel Mouafo; Boudjeko, Thaddée; Simao-Beaunoir, Anne-Marie; Lerat, Sylvain; Tsala, Éric; Monga, Ernest; Beaulieu, Carole

    2016-01-01

    Vascular plants are commonly colonized by endophytic actinobacteria. However, very little is known about the relationship between these microorganisms and cacao fruits. In order to determine the physiological and taxonomic relationships between the members of this community, actinobacteria were isolated from cacao fruits and seeds. Among the 49 isolates recovered, 11 morphologically distinct isolates were selected for further characterization. Sequencing of the 16S rRNA gene allowed the partition of the selected isolates into three phylogenetic clades. Most of the selected endophytic isolates belonged to the Streptomyces violaceusniger clade. Physiological characterization was carried out and a similarity index was used to cluster the isolates. However, clustering based on physiological properties did not match phylogenetic lineages. Isolates were also characterized for traits commonly associated with plant growth-promoting bacteria, including antibiosis and auxin biosynthesis. All isolates exhibited resistance to geldanamycin, whereas only two isolates were shown to produce this antibiotic. Endophytes were inoculated on radish seedlings and most isolates were found to possess plant growth-promoting abilities. These endophytic actinobacteria inhibited the growth of various plant pathogenic fungi and/or bacteria. The present study showed that S. violaceusniger clade members represent a significant part of the actinobacterial community living as endophytes in cacao fruits and seeds. While several members of this clade are known to be geldanamycin producers and efficient biocontrol agents of plant diseases, we herein established the endophytic lifestyle of some of these microorganisms, demonstrating their potential as plant health agents.

  1. An endophytic Basidiomycete, Grammothele lineata, isolated from Corchorus olitorius, produces paclitaxel that shows cytotoxicity.

    PubMed

    Das, Avizit; Rahman, Mohammad Imtiazur; Ferdous, Ahlan Sabah; Amin, Al-; Rahman, Mohammad Mahbubur; Nahar, Nilufar; Uddin, Md Aftab; Islam, Mohammad Riazul; Khan, Haseena

    2017-01-01

    Grammothele lineata, an endophyte isolated in our laboratory from jute (Corchorus olitorius acc. 2015) was found to be a substantial paclitaxel producer. Taxol and its related compounds, produced by this endophyte were extracted by growing the fungus in simple nutrient media (potato dextrose broth, PDB). Taxol was identified and characterized by different analytical techniques (TLC, HPLC, FTIR, LC-ESI-MS/MS) following its extraction by ethyl acetate. In PDB media, this fungus was found to produce 382.2 μgL-1 of taxol which is about 7.6 x103 fold higher than the first reported endophytic fungi, Taxomyces andreanae. The extracted taxol exhibited cytotoxic activity in an in vitro culture of HeLa cancer cell line. The fungal extract also exhibited antifungal and antibacterial activities against different pathogenic strains. This is the first report of a jute endophytic fungus harboring the capacity to produce taxol and also the first reported taxol producing species that belongs to the Basidiomycota phylum, so far unknown to be a taxol producer. These findings suggest that the fungal endophyte, Grammothele lineata can be an excellent source of taxol and can also serve as a potential species for chemical and genetic engineering to enhance further the production of taxol.

  2. Degradation of the polyamine alkaloid aphelandrine by endophytic fungi isolated from Aphelandra tetragona.

    PubMed

    Werner, C; Petrini, O; Hesse, M

    1997-10-15

    Members of the genus Aphelandra (Acanthaceae) produce rare macrocyclic polyamine alkaloids which consist of spermine acylated with two units of 3-(4-hydroxyphenyl)prop-2-enoic acid. Endophytic fungi were isolated from roots and shoots of Aphelandra tetragona and tested for their ability to metabolize the main alkaloid aphelandrine, which accumulates exclusively in the roots of the plants. Several endophytes were able to metabolize aphelandrine but only root endophytes belonging to the Nectriaceae were good metabolizers. In addition, the endophytes were grown on an agar medium containing putrescine, spermidine, or spermine as the sole nitrogen source. All fungi were able to grow on putrescine, but only the good aphelandrine metabolizers grew well on spermidine or spermine. Acremonium sp. 15, one of the most active metabolizers, grew also on a medium containing aphelandrine as sole nitrogen source. A number of strains thought to be conspecific with Acremonium sp. 15 were also tested for their ability to metabolize aphelandrine. The ability of the endophytes to metabolize aphelandrine suggests an ecological adaptation of the symbionts to their host. The possibility of using the aphelandrine metabolism as a taxonomic character is briefly discussed.

  3. Genetic diversity and antimicrobial activity of endophytic Myrothecium spp. isolated from Calophyllum apetalum and Garcinia morella.

    PubMed

    Ruma, Karmakar; Sunil, Kumar; Kini, Kukkundoor R; Prakash, Harischandra Sripathy

    2015-11-01

    Calophyllum apetalum and Garcinia morella, medicinal plants are endemic to Western Ghats, Karnataka, India. Sixteen Myrothecium isolates were obtained from the tissues of bark and twigs of these plants. The purpose of this study was to explore the antimicrobial activity and genetic variability of the endophytic Myrothecium isolates. The antimicrobial activity as well as the genetic diversity of endophytic Myrothecium species was investigated through RAPD, ISSR and ITS sequence analysis. Myrothecium isolates were genotypically compared by RAPD and ISSR techniques, 510 and 189 reproducible polymorphic bands were obtained using 20 RAPD and ten ISSR primers respectively. The isolates grouped into four main clades and subgroups using unweighted pair group method with arithmetic mean cluster analysis. rDNA ITS sequence analysis presented better resolution for characterising the isolates of Myrothecium spp. The clustering patterns of the isolates were almost similar when compared with RAPD and ISSR dendograms. The results signify that RAPD, ISSR and ITS analysis can be employed to distinguish the genetic diversity of the Myrothecium species. The endophytic and pathogenic strains were compared by maximum parsimony, maximum likelihood and neighbour joining methods. One isolate (JX862206) amongst the 16 Myrothecium isolates exhibited potent antibacterial and as well as anti-Candida activity.

  4. Phylogenetic and chemical diversity of fungal endophytes isolated from Silybum marianum (L) Gaertn. (milk thistle)

    PubMed Central

    Raja, Huzefa A.; Kaur, Amninder; El-Elimat, Tamam; Figueroa, Mario; Kumar, Rahul; Deep, Gagan; Agarwal, Rajesh; Faeth, Stanley H.; Cech, Nadja B.; Oberlies, Nicholas H.

    2015-01-01

    Use of the herb milk thistle (Silybum marianum) is widespread, and its chemistry has been studied for over 50 years. However, milk thistle endophytes have not been studied previously for their fungal and chemical diversity. We examined the fungal endophytes inhabiting this medicinal herb to determine: (1) species composition and phylogenetic diversity of fungal endophytes; (2) chemical diversity of secondary metabolites produced by these organisms; and (3) cytotoxicity of the pure compounds against the human prostate carcinoma (PC-3) cell line. Forty-one fungal isolates were identified from milk thistle comprising 25 operational taxonomic units based on BLAST search via GenBank using published authentic sequences from nuclear ribosomal internal transcribed spacer sequence data. Maximum likelihood analyses of partial 28S rRNA gene showed that these endophytes had phylogenetic affinities to four major classes of Ascomycota, the Dothideomycetes, Sordariomycetes, Eurotiomycetes, and Leotiomycetes. Chemical studies of solid–substrate fermentation cultures led to the isolation of four new natural products. In addition, 58 known secondary metabolites, representing diverse biosynthetic classes, were isolated and characterized using a suite of nuclear magnetic resonance and mass spectrometry techniques. Selected pure compounds were tested against the PC-3 cell line, where six compounds displayed cytotoxicity. PMID:26000195

  5. Morphological and genetic characterization of endophytic bacteria isolated from roots of different maize genotypes.

    PubMed

    Ikeda, Angela Cristina; Bassani, Luciana Lange; Adamoski, Douglas; Stringari, Danyelle; Cordeiro, Vanessa Kava; Glienke, Chirlei; Steffens, Maria Berenice Reynaud; Hungria, Mariangela; Galli-Terasawa, Lygia Vitoria

    2013-01-01

    Maize is one of the most important crops worldwide, and in Brazil, the state of Paraná stands as its largest producer. The crop demands high inputs of N fertilizers, therefore all strategies aiming to optimize the grain production with lower inputs are very relevant. Endophytic bacteria have a high potential to increment maize grain yield by means of input via biological nitrogen fixation and/or plant growth promotion, in this last case increasing the absorption of water and nutrients by the plants. In this study, we established a collection of 217 endophytic bacteria, isolated from roots of four lineages and three hybrid genotypes of maize, and isolated in four different N-free culture media. Biochemical-comprising growth in different carbon sources, intrinsic tolerance to antibiotics, and biochemical tests for catalase, nitrate reductase, urease, and growth in N-free media in vitro-and genetic characterization by BOX-PCR revealed great variability among the isolates. Both commercial hybrids and homozygous lineages were broadly colonized by endophytes, and sequencing of the 16S rRNA gene revealed the presence of bacteria belonging to the genera Pantoea, Bacillus, Burkholderia, and Klebsiella. Qualitative differences in endophytic colonization were detected between lineages and hybrid genotypes.

  6. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell (Scrophulariaceae)

    PubMed Central

    2014-01-01

    Background Endophytes, which reside in plant tissues, have the potential to produce novel metabolites with immense benefits for health industry. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell were investigated. Methods Endophytic fungi were isolated from the Bacopa monnieri. Extracts from liquid cultures were tested for cytotoxicity against a number of cancer cell lines using the MTT assay. Antimicrobial activity was determined using the micro dilution method. Results 22% of the examined extracts showed potent (IC50 of <20 μg/ml) cytotoxic activity against HCT-116 cell line. 5.5%, 11%, 11% of the extracts were found to be cytotoxic for MCF-7, PC-3, and A-549 cell lines respectively. 33% extracts displayed antimicrobial activity against at least one test organism with MIC value 10–100 μg/ml. The isolate B9_Pink showed the most potent cytotoxic activity for all the cell lines examined and maximum antimicrobial activity against the four pathogens examined which was followed by B19. Conclusions Results indicated the potential for production of bioactive agents from endophytes of Bacopa monnieri. PMID:24512530

  7. Characterization of Antifungal Natural Products Isolated from Endophytic Fungi of Finger Millet (Eleusine coracana).

    PubMed

    Mousa, Walaa Kamel; Schwan, Adrian L; Raizada, Manish N

    2016-09-03

    Finger millet is an ancient African-Indian crop that is resistant to many pathogens including the fungus, Fusarium graminearum. We previously reported the first isolation of putative fungal endophytes from finger millet and showed that the crude extracts of four strains had anti-Fusarium activity. However, active compounds were isolated from only one strain. The objectives of this study were to confirm the endophytic lifestyle of the three remaining anti-Fusarium isolates, to identify the major underlying antifungal compounds, and to initially characterize the mode(s) of action of each compound. Results of confocal microscopy and a plant disease assay were consistent with the three fungal strains behaving as endophytes. Using bio-assay guided fractionation and spectroscopic structural elucidation, three anti-Fusarium secondary metabolites were purified and characterized. These molecules were not previously reported to derive from fungi nor have antifungal activity. The purified antifungal compounds were: 5-hydroxy 2(3H)-benzofuranone, dehydrocostus lactone (guaianolide sesquiterpene lactone), and harpagoside (an iridoide glycoside). Light microscopy and vitality staining were used to visualize the in vitro interactions between each compound and Fusarium; the results suggested a mixed fungicidal/fungistatic mode of action. We conclude that finger millet possesses fungal endophytes that can synthesize anti-fungal compounds not previously reported as bio-fungicides against F. graminearum.

  8. Antimicrobial activities of endophytic fungi obtained from the arid zone invasive plant Opuntia dillenii and the isolation of equisetin, from endophytic Fusarium sp.

    PubMed

    Ratnaweera, Pamoda B; de Silva, E Dilip; Williams, David E; Andersen, Raymond J

    2015-07-10

    Opuntia dillenii is an invasive plant well established in the harsh South-Eastern arid zone of Sri Lanka. Evidence suggests it is likely that the endophytic fungal populations of O. dillenii assist the host in overcoming biotic and abiotic stress by producing biologically active metabolites. With this in mind there is potential to discover novel natural products with useful biological activities from this hitherto poorly investigated source. Consequently, an investigation of the antimicrobial activities of the endophytes of O. dillenii, that occupies a unique ecological niche, may well provide useful leads in the discovery of new pharmaceuticals. Endophytic fungi were isolated from the surface sterilized cladodes and flowers of O. dillenii using several nutrient media and the antimicrobial activities were evaluated against three Gram-positive and two Gram-negative bacteria and Candida albicans. The two most bioactive fungi were identified by colony morphology and DNA sequencing. The secondary metabolite of the endophyte Fusarium sp. exhibiting the best activity was isolated via bioassay guided chromatography. The chemical structure was elucidated from the ESIMS and NMR spectroscopic data obtained for the active metabolite. The minimum inhibitory concentrations (MICs) of the active compound were determined. Eight endophytic fungi were isolated from O. dillenii and all except one showed antibacterial activities against at least one of the test bacteria. All extracts were inactive against C. albicans. The most bioactive fungus was identified as Fusarium sp. and the second most active as Aspergillus niger. The structure of the major antibacterial compound of the Fusarium sp. was shown to be the tetramic acid derivative, equisetin. The MIC's for equisetin were 8 μg mL(-1) against Bacillus subtilis, 16 μg mL(-1) against Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus (MRSA). O. dillenii, harbors several endophytic fungi capable of producing

  9. Screening of endophytic bacteria isolated from leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] for cytokinin-like compounds

    PubMed Central

    Bhore, Subhash J; Ravichantar, Nithya; Loh, Chye Ying

    2010-01-01

    Endophytic bacteria are harmless in most plant species; and known to boost the growth and development of the host plants probably by secreting growth hormones. The isolation, identification and screening of endophytic bacteria for the plant growth regulators like cytokinin are needed to get the leads for their applications in agriculture sector. We describe the isolation and identification of the bacterial endophytes from the leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] and their screening for cytokinin-like compounds. We isolated three endophytic bacteria from the leaves of G. procumbens collected from the forest research institute of Malaysia (FRIM). They were further identified using amplified 16S rRNA gene sequence based method of bacterial identification. The ethyl acetate extracts of the isolates-broth were analyzed using cucumber cotyledon greening bioassay (CCGB) to determine the presence of cytokinin-like compounds. Consequently, the bacterial putative endophytes were identified as Psuedomonas resinovorans, Paenibacillus polymaxa, and Acenitobacter calcoaceticus. Broth-extracts from two (Psuedomonas resinovorans and Paenibacillus polymaxa) of the three putative bacterial endophytes show the positive results in their screening for cytokinin-like compounds using CCGB. Thus, we hypothesize that the bacterial putative endophytes of G. procumbens that produce cytokinin-like compounds might have a role in the growth and development of G. procumbens. Abbreviations CCGB - Cucumber cotyledon greening bioassay, rDNA - Ribosomal DNA, K12, BAP - 6-Benzylaminopurine, Db1, MSA - Multiple sequence alignment. 8081, PMID:21364796

  10. Endophytic fungi isolated from oil-seed crop Jatropha curcas produces oil and exhibit antifungal activity.

    PubMed

    Kumar, Susheel; Kaushik, Nutan

    2013-01-01

    Jatropha curcas L., a perennial plant grown in tropics and subtropics is popularly known for its potential as biofuel. The plant is reported to survive under varying environmental conditions having tolerance to stress and an ability to manage pest and diseases. The plant was explored for its endophytic fungi for use in crop protection. Endophytic fungi were isolated from leaf of Jatropha curcas, collected from New Delhi, India. Four isolates were identified as Colletotrichum truncatum, and other isolates were identified as Nigrospora oryzae, Fusarium proliferatum, Guignardia cammillae, Alternaria destruens, and Chaetomium sp. Dual plate culture bioassays and bioactivity assays of solvent extracts of fungal mycelia showed that isolates of Colletotrichum truncatum were effective against plant pathogenic fungi Fusarium oxysporum and Sclerotinia sclerotiorum. Isolate EF13 had highest activity against S. sclerotiorum. Extracts of active endophytic fungi were prepared and tested against S. sclerotiorum. Ethyl acetate and methanol extract of C. truncatum EF10 showed 71.7% and 70% growth inhibition, respectively. Hexane extracts of C. truncatum isolates EF9, EF10, and EF13 yielded oil and the oil from EF10 was similar to oil of the host plant, i.e., J. curcas.

  11. Endophytic Fungi Isolated from Oil-Seed Crop Jatropha curcas Produces Oil and Exhibit Antifungal Activity

    PubMed Central

    Kumar, Susheel; Kaushik, Nutan

    2013-01-01

    Jatropha curcas L., a perennial plant grown in tropics and subtropics is popularly known for its potential as biofuel. The plant is reported to survive under varying environmental conditions having tolerance to stress and an ability to manage pest and diseases. The plant was explored for its endophytic fungi for use in crop protection. Endophytic fungi were isolated from leaf of Jatropha curcas, collected from New Delhi, India. Four isolates were identified as Colletotrichum truncatum, and other isolates were identified as Nigrospora oryzae, Fusarium proliferatum, Guignardia cammillae, Alternaria destruens, and Chaetomium sp. Dual plate culture bioassays and bioactivity assays of solvent extracts of fungal mycelia showed that isolates of Colletotrichum truncatum were effective against plant pathogenic fungi Fusarium oxysporum and Sclerotinia sclerotiorum. Isolate EF13 had highest activity against S. sclerotiorum. Extracts of active endophytic fungi were prepared and tested against S. sclerotiorum. Ethyl acetate and methanol extract of C. truncatum EF10 showed 71.7% and 70% growth inhibition, respectively. Hexane extracts of C. truncatum isolates EF9, EF10, and EF13 yielded oil and the oil from EF10 was similar to oil of the host plant, i.e., J. curcas. PMID:23409154

  12. Isolation and morphological and metabolic characterization of common endophytes in annually burned tallgrass prairie.

    PubMed

    Mandyam, Keerthi; Loughin, Thomas; Jumpponen, Ari

    2010-01-01

    Dark septate endophytes (DSE) are common and abundant root-colonizing fungi in the native tallgrass prairie. To characterize DSE fungi were isolated from roots of mixed tallgrass prairie plant communities. Isolates were grouped according to morphology, and the grouping was refined by ITS-RFLP and/or sequencing of the ITS region. Sporulating species of Periconia, Fusarium, Microdochium and Aspergillus were isolated along with many sterile fungi. Leek resynthesis was used to quickly screen for DSE fungi among the isolates. Periconia macro-spinosa and Microdochium sp. formed typical DSE structures in the roots; Periconia produced melanized intracellular microsclerotia in host root cortex, whereas Microdochium produced abundant melanized inter- and intracellular chlamydospores. To further validate the results of the leek resynthesis growth responses of leek and a dominant prairie grass, Andropogon gerardii, were assessed in a laboratory resynthesis system. Leek growth mainly was unresponsive to the inoculation with Periconia or Microdochium, whereas Andropogon tended to respond positively. Select Periconia and Microdochium isolates were tested further for their enzymatic capabilities and for ability to use organic and inorganic nitrogen sources. These fungi tested positive for amylase, cellulase, polyphenol oxidases and gelatinase. Periconia isolates used both organic and inorganic nitrogen sources. Our study identified distinct endophytes in a tallgrass prairie ecosystem and indicated that these endo-phytes can use a variety of complex nutrient sources, suggesting facultative biotrophic and saprotrophic habits.

  13. Fungal endophytes isolated from Protium heptaphyllum and Trattinnickia rhoifolia as antagonists of Fusarium oxysporum.

    PubMed

    Fierro-Cruz, Juan E; Jiménez, Pedro; Coy-Barrera, Ericsson

    Control of fungal pathogens is mainly addressed by the use of chemically synthesized fungicides which result in environmental pollution, developing resistance after prolonged use. In this context, endophytes have been recognized as potential biocontrollers, and also as a promising source of antifungal metabolites. Therefore, as part of our research on phytopathogen controllers, 355 fungal endophytes were isolated from Protium heptaphyllum and Trattinnickia rhoifolia (Burseraceae), both ethnobotanically important tree species that produce secondary metabolites of agronomic and industrial interest. Endophytes were tested by in vitro dual culture against Fusarium oxysporum, a phytopathogen of agronomic importance. Five endophytes exerted at least 40% inhibition on F. oxysporum growth. Ethyl acetate (EtOAc) extracts were obtained from the most active antagonistic fungi, after growing them in three different liquid media. The extracts were tested against a conidial suspension of F. oxysporum by direct bioautography. Two extracts derived from fungi identified as Chaetomium globosum, F211_UMNG and Meyerozima sp. F281_UMNG showed inhibition of pathogen growth. Isolate C. globosum, F211_UMNG was selected for a chemical analysis by RP-HPLC-DAD-ESI-MS and antifungal molecules such as cladosporin, chaetoatrosin A and chaetoviridin A were annotated and identified based on their MS data. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  14. Saccharopolyspora subtropica sp. nov., a thermophilic actinomycete isolated from soil of a sugar cane field.

    PubMed

    Wu, Hao; Liu, Bin; Pan, Shangli

    2016-05-01

    A novel thermophilic actinomycete, designated strain T3T, was isolated from a soil sample of a sugar cane field. The strain grew at 25-60 °C (optimum 37-50 °C), at pH 6.0-11.0 (optimum 7.0-9.0) and with 0-12.0 % (w/v) NaCl (optimum 0-7 %). The aerial mycelium was white and the vegetative mycelium was colourless to pale yellow. The substrate mycelium fragmented into rod-shaped elements after 4-5 days at 50 °C. The aerial mycelium formed flexuous chains of 5-20 spores per chain; the oval-shaped spores had spiny surfaces and were non-motile. The organism contained meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The whole-cell sugars consisted of arabinose, galactose and ribose. The cellular fatty acid profile consisted mainly of anteiso-C17 : 0, iso-C17 : 0 and iso-C16 : 0. The quinone system was composed predominantly of MK-9(H4). The phospholipids detected were diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylmethylethanolamine and ninhydrin-positive glycophospholipids. The DNA G+C content of strain T3T was 71.3 mol%. The organism showed a combination of morphological and chemotaxonomic properties typical of members of the genus Saccharopolyspora. In the 16S rRNA gene tree of Saccharopolyspora it formed a distinct phyletic line and was related most closely to Saccharopolyspora thermophila 216T. However, the phenotypic characteristics of strain T3T were significantly different from those of S. thermophila 216T and DNA-DNA hybridization revealed a low level of relatedness (28.6-32.3 %) between them. Based on the phenotypic and phylogenetic data, strain T3T represents a novel species in the genus Saccharopolyspora, for which the name Saccharopolyspora subtropica sp. nov. is proposed. The type strain is T3T ( = DSM 46801T = CGMCC 4.7206T).

  15. Antioxidant treatments counteract the non-culturability of bacterial endophytes isolated from legume nodules.

    PubMed

    Muresu, Rosella; Tondello, Alessandra; Polone, Elisa; Sulas, Leonardo; Baldan, Barbara; Squartini, Andrea

    2013-06-01

    In many wild legumes, attempts to cultivate nodule bacteria fail. We hypothesized that the limited culturability could be related to injury from oxidative stress caused by disruption of plant tissues during isolation. To test that, we isolated bacteria from nodules of Hedysarum spinosissimum and Tetragonolobus purpureus using buffers supplemented with scavenging systems to prevent damage from reactive oxygen species (ROS). Treatments included the following: antioxidants (glutathione, ascorbate, EDTA) or enzymes (catalase, peroxidase, superoxide dismutase), tested either as modified squashing buffers or added in plates. Some combinations yielded dramatic increases of culturability. Different endophytes were found, including additional Rhizobiaceae that were not the primary symbiont and were unable to nodulate. Their H2O2 tolerance in broth culture showed differences consistent with the unequal culturability observed. In wild legumes species, ROS generation during extraction appears to be a major factor limiting microbiota isolation, and protocols presented here significantly improve the recovery of culturable bacterial endophytes from plants.

  16. Endophytic fungi from Dragon's blood specimens: isolation, identification, phylogenetic diversity and bioactivity.

    PubMed

    Cui, Jin-long; Guo, Shun-xing; Dong, Hailing; Xiao, Peigen

    2011-08-01

    Endophytic fungi from Dragon's blood specimens of different locations of China were characterized taxonomically and investigated concerning their antimicrobial and antitumor activity against six pathogenic microbes and five tumor cells. A total of 49 endophytic fungi were obtained from Dragon's blood materials of Dracaena spp., 18 taxa were represented by 43 (87.8%) isolates and only six (12.2%) isolates were unknown. Twenty (40.8%) of the isolates displayed antimicrobial activity against at least one pathogenic microorganism. Three isolates YNDC07, BJDC06 and BJDC09 displayed significant antimicrobial activities against Staphylococcus aureus, Cryptococcus neoformans and Aspergillus fumigates, respectively. The results of antitumor activity by the MTT assay revealed that 26.5%, 69.4%, 48.9%, 6.1% and 42.9% of isolate fermentation broths displayed growth inhibition on HepG2 cells, SKVO3 cells, MCF7 cells, HL-60 cells and 293-T cells, respectively. HNDC09 and HNDC10 showed very strong antitumor activity against MCF7 and 293-T, respectively. The results showed that endophytic fungi in Dragon's blood samples were valuable in screening antitumor and antimicrobial bioactivity agents.

  17. Petroleum degradation by endophytic Streptomyces spp. isolated from plants grown in contaminated soil of southern Algeria.

    PubMed

    Baoune, Hafida; Ould El Hadj-Khelil, Aminata; Pucci, Graciela; Sineli, Pedro; Loucif, Lotfi; Polti, Marta Alejandra

    2017-09-15

    Petroleum hydrocarbons are well known by their high toxicity and recalcitrant properties. Their increasing utilization around worldwide led to environmental contamination. Phytoremediation using plant-associated microbe is an interesting approach for petroleum degradation and actinobacteria have a great potential for that. For this purpose, our study aimed to isolate, characterize, and assess the ability of endophytic actinobacteria to degrade crude petroleum, as well as to produce plant growth promoting traits. Seventeen endophytic actinobacteria were isolated from roots of plants grown naturally in sandy contaminated soil. Among them, six isolates were selected on the basis of their tolerance to petroleum on solid minimal medium and characterized by 16S rDNA gene sequencing. All petroleum-tolerant isolates belonged to the Streptomyces genus. Determination by crude oil degradation by gas chromatorgraph-flame ionization detector revealed that five strains could use petroleum as sole carbon and energy source and the petroleum removal achieved up to 98% after 7 days of incubation. These isolates displayed an important role in the degradation of the n-alkanes (C6-C30), aromatic and polycyclic aromatic hydrocarbons. All strains showed a wide range of plant growth promoting features such as siderophores, phosphate solubilization, 1-aminocyclopropane-1-carboxylate deaminase, nitrogen fixation and indole-3-acetic acid production as well as biosurfactant production. This is the first study highlighting the petroleum degradation ability and plant growth promoting attributes of endophytic Streptomyces. The finding suggests that the endophytic actinobacteria isolated are promising candidates for improving phytoremediation efficiency of petroleum contaminated soil. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Isolation, Taxonomy, and Antagonistic Properties of Halophilic Actinomycetes in Saharan Soils of Algeria ▿

    PubMed Central

    Meklat, Atika; Sabaou, Nasserdine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed

    2011-01-01

    The diversity of a population of 52 halophilic actinomycetes was evaluated by a polyphasic approach, which showed the presence of members of the Actinopolyspora, Nocardiopsis, Saccharomonospora, Streptomonospora, and Saccharopolyspora genera. One strain was considered to be a new member of the last genus, and several other strains seemed to be new species. Furthermore, 50% of strains were active against a broad range of indicators and contained genes encoding polyketide synthetases and nonribosomal peptide synthetases. PMID:21764956

  19. Isolation, taxonomy, and antagonistic properties of halophilic actinomycetes in Saharan soils of Algeria.

    PubMed

    Meklat, Atika; Sabaou, Nasserdine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed

    2011-09-01

    The diversity of a population of 52 halophilic actinomycetes was evaluated by a polyphasic approach, which showed the presence of members of the Actinopolyspora, Nocardiopsis, Saccharomonospora, Streptomonospora, and Saccharopolyspora genera. One strain was considered to be a new member of the last genus, and several other strains seemed to be new species. Furthermore, 50% of strains were active against a broad range of indicators and contained genes encoding polyketide synthetases and nonribosomal peptide synthetases.

  20. [Study of marine actinomycetes isolated from the central coast of Peru and their antibacterial activity against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis].

    PubMed

    León, Jorge; Aponte, Juan José; Rojas, Rosario; Cuadra, D'Lourdes; Ayala, Nathaly; Tomás, Gloria; Guerrero, Marco

    2011-06-01

    To determine the antimicrobial potential of marine actinomycetes against drug-resistant pathogens represented by strains of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE). Strains of actinomycetes (29) isolated from marine sediment were evaluated by their characteristics in two culture media and by testing their inhibitory capacity by in vitro antagonism against multi-drug resistant (MDR) pathogenic bacteria for MRSA and VRE. Organic extracts of 3 selected actinomicetes were processed to determine the minimum inhibitory concentration (MIC) of the active compound. Most isolated actinomycetes belong to a homogeneous group of write-gray actinomycetes with a good growth in Marine Agar. The inhibitory rates of the isolates were above 85% for both pathogens with inhibition zones greater than 69 and 78 mm in diameter for MRSA and VRE respectively. Dichloromethane extracts of 3 isolates (I-400A, B1-T61, M10-77) showed strong inhibitory activity of both pathogens, M10-77 being the highest actinomycete strain with antibiotic activity against methicillin-resistant S. aureus ATCC 43300 and vancomycin-resistant E. faecalis ATCC 51299 with a minimum inhibitory concentrations (MIC) of 7.9 and 31.7 μg/ml respectively. Phylogenetic analysis of M10-77 strain showed 99% similarity with the marine species Streptomyces erythrogriseus. Marine sediments of the central coast of Peru, are a source of actinomycetes strains showing high capacity to produce bioactive compounds able to inhibit pathogens classified as multi-drug-resistant such as methicillin-resistant S. aureus and vancomycin-resistant E. faecalis.

  1. Lethality of cytochalasin B and other compounds isolated from fungus Aspergillus sp. (Trichocomaceae) endophyte of Bauhinia guianensis (Fabaceae).

    PubMed

    Feitosa, André de O; Dias, Amanda Cristina S; Ramos, Gisele da C; Bitencourt, Heriberto R; Siqueira, José Edson S; Marinho, Patrícia Santana B; Barison, Andersson; Ocampos, Fernanda M M; Marinho, Andrey Moacir do R

    Endophytic fungi are fungi that colonize internal tissues of plants; several biologically active compounds have been isolated from these fungi. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, this study aimed the isolation and structural identification of ergosterol (1), ergosterol peroxide (2), mevalonolactone (3), cytochalasin B (4) and cytochalasin H (5) from Aspergillus sp. EJC 04, an endophytic fungus from Bauhinia guianensis. The cytochalasin B (4) and the diacetate derivative of cytochalasin B (4a) showed high lethality in the brine shrimp assay. This is the first occurrence of cytochalasins in Amazonian endophytic fungi from B. guianensis. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  2. Indentification of vincamine indole alkaloids producing endophytic fungi isolated from Nerium indicum, Apocynaceae.

    PubMed

    Na, Ren; Jiajia, Liu; Dongliang, Yang; Yingzi, Peng; Juan, Hong; Xiong, Liu; Nana, Zhao; Jing, Zhou; Yitian, Luo

    2016-11-01

    Vincamine, a monoterpenoid indole alkaloid which had been marketed as nootropic drugs for the treatment of cerebral insufficiencies, is widely found in plants of the Apocynaceae family. Nerium indicum is a plant belonging to the Apocynaceae family. So, the purpose of this research was designed to investigate the vincamine alkaloids producing endophytic fungi from Nerium indicum, Apocynaceae. 11 strains of endophytic fungi, isolated from the stems and roots of the plant, were grouped into 5 genera on the basis of morphological characteristics. All fungal isolates were fermented and their extracts were preliminary screened by Dragendorff's reagent and thin layer chromatography (TLC). One isolated strain CH1, isolated from the stems of Nerium indicum, had the same Rf value (about 0.56) as authentic vincamine. The extracts of strain CH1 were further analyzed by high performance liquid chromatography (HPLC) and liquid chromatograph-mass spectrometry (LC-MS), and the results showed that the strain CH1 could produce vincamine and vincamine analogues. The acetylcholinesterase (AchE) inhibitory activity assays using Ellman's method revealed that the metabolites of strain CH1 had significant AchE inhibitory activity with an IC50 value of 5.16μg/mL. The isolate CH1 was identified as Geomyces sp. based on morphological and molecular identification, and has been deposited in the China Center for Type Culture Collection (CCTCCM 2014676). This study first reported the natural compounds tabersonine and ethyl-vincamine from endophytic fungi CH1, Geomyces sp. In conclusion, the fungal endophytes from Nerium indicum can be used as alternative source for the production of vincamine and vincamine analogues. Copyright © 2016 Elsevier GmbH. All rights reserved.

  3. Isolation, Characterization, and Insecticidal Activity of an Endophyte of Drunken Horse Grass, Achnatherum inebrians

    PubMed Central

    Shi, YingWu; Zhang, Xuebing; Lou, Kai

    2013-01-01

    Endophytic microorganisms reside within plant tissues and have often been found to promote plant growth. In this study, endophytic microorganisms were isolated from the roots, stems, leaves, and seeds of healthy drunken horse grass, Achnatherum inebrians (Hance) Keng (Poales: Poaceae), through the use of a grinding separation method and identified by a dual approach of morphological and physiological observation and 16S rRNA gene-based (for bacteria) and internal transcribed sequence-based (for fungi) molecular identification. The endophytes were then inoculated into liquid media for fermentation, and their crude extracts were employed for insecticidal activity tests using slide disc immersion and nebulization methods. A total of 89 bacteria species, which were classified into eight genera, Bacillus, Pseudomonas, Actinomyces, Corynebacterium, Acinetobacter, Sphingomonas, Paenibacillus, and Phyllobacterium, and two fungi, Claviceps and Chaetomium, were isolated. Of these species, isolates Streptomyces albus (Rossi-Doria) Waksman and Henrici (Actinomycetales: Streptomycetaceae) (GA) and Claviceps purpurea (Fr.) Tul. (Hypocreales: Clavicipitaceae) (PF-2) were shown to produce mortality rates of more than 90% in the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), after first and second screenings. The isolates PF-2 and GA associated with A. inebrians had significant insecticidal activities towards A. gossypii Glover (Hemiptera: Aphididae) and may provide a new biological resource for exploring a new microbial insecticide. PMID:24784492

  4. Screening of endophytic Streptomycetes isolated from Parthenium hysterophorus L. against nosocomial pathogens.

    PubMed

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida

    2013-03-01

    Parthenium hysterophorus L. is an obnoxious weed of the family asteraceae recognized for its detrimental effects and significant economic losses to agriculture. In this study 42 endophytic streptomycetes strains were isolated from its roots and leaves. The isolates were identified by morphological, microscopic, biochemical and physiological characterization as members of genus Streptomyces. In 16S rRNA gene sequencing the selected isolates exhibited maximum similarity with Streptomyces rochei (99%), Streptomyces litmocidini (99%), Streptomyces enissocaesili (99%), Streptomyces djakartensis (99%), Streptomyces olivaceus (99%), Streptomyces spp (99%), Streptomyces plicatus (99%), Streptomyces geysiriensis (99%) and Streptomyces vinaceusdrappus (99%). In biological screening the crude extracts of 12 strains exhibited significant antimicrobial activity against multi drug resistant nosocomial pathogens including Pseudomonas, Enterobacter, Bacillus, Escherichia coli, Staphlococcus aureus and Candida albicans. In chemical screening by thin layer chromatography (TLC) the extracts exhibited an impressive diversity of the bioactive secondary metabolites. Additionally high performance liquid chromatography (HPLC-UV) chromatographs revealed many impressive peaks of unidentified bioactive metabolites. As such this is a first study reporting the isolation, identification and screening of endophytic Streptomyces from the invasive weed. The results provide an insight into an untapped endophytic environment yet to be explored which might be a promising source of lead antimicrobial agents.

  5. Isolation, characterization, and insecticidal activity of an endophyte of drunken horse grass, Achnatherum inebrians.

    PubMed

    Shi, YingWu; Zhang, Xuebing; Lou, Kai

    2013-01-01

    Abstract Endophytic microorganisms reside within plant tissues and have often been found to promote plant growth. In this study, endophytic microorganisms were isolated from the roots, stems, leaves, and seeds of healthy drunken horse grass, Achnatherum inebrians (Hance) Keng (Poales: Poaceae), through the use of a grinding separation method and identified by a dual approach of morphological and physiological observation and 16S rRNA gene-based (for bacteria) and internal transcribed sequence-based (for fungi) molecular identification. The endophytes were then inoculated into liquid media for fermentation, and their crude extracts were employed for insecticidal activity tests using slide disc immersion and nebulization methods. A total of 89 bacteria species, which were classified into eight genera, Bacillus, Pseudomonas, Actinomyces, Corynebacterium, Acinetobacter, Sphingomonas, Paenibacillus, and Phyllobacterium, and two fungi, Claviceps and Chaetomium, were isolated. Of these species, isolates Streptomyces albus (Rossi-Doria) Waksman and Henrici (Actinomycetales: Streptomycetaceae) (GA) and Claviceps purpurea (Fr.) Tul. (Hypocreales: Clavicipitaceae) (PF-2) were shown to produce mortality rates of more than 90% in the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), after first and second screenings. The isolates PF-2 and GA associated with A. inebrians had significant insecticidal activities towards A. gossypii Glover (Hemiptera: Aphididae) and may provide a new biological resource for exploring a new microbial insecticide.

  6. Piperine production by endophytic fungus Colletotrichum gloeosporioides isolated from Piper nigrum.

    PubMed

    Chithra, S; Jasim, B; Sachidanandan, P; Jyothis, M; Radhakrishnan, E K

    2014-03-15

    Many endophytic fungi have been reported with the biosynthetic potential to produce same or similar metabolites present in host plants. The adaptations that might have acquired by these fungi as a result of the long-term association with their host plants can be the possible basis of their biosynthetic potential. The bioactive compounds originated from endophytes are currently explored for their potential applications in pharmaceutical, agriculture and food industries. Piper nigrum, a plant of the Piperaceae is very remarkable because of the presence of the alkaloid piperine. Piperine has been reported to have broad bioactive properties ranging from antimicrobial, antidepressant, anti-inflammatory, antioxidative to anticancer activities. Interestingly, piperine also plays a vital role in increasing the bioavailability of many drugs which again is a promising property. The current study was carried out to identify piperine producing endophytic fungus from Piper nigrum L. By screening various endophytic fungi, the isolate which was identified as member of Colletotrichum gloeosporioides was found to have the ability to form piperine and was confirmed by HPLC and LCMS. Considering the broad bioactive potential of piperine, the piperine producing fungi identified in the study can expect to have much industrial potential. Copyright © 2013 Elsevier GmbH. All rights reserved.

  7. Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves.

    PubMed

    Singh, Ankit Kumar; Sharma, Rajesh Kumar; Sharma, Varsha; Singh, Tanmay; Kumar, Rajesh; Kumari, Dimple

    2017-05-01

    The objective of this study was to isolate endophytic bacteria from Azadirachta indica (neem) leaves, their identification and investigate their antibacterial activity against three Gram-positive bacteria, Staphylococcus aureus, Streptococcus pyogenes and Bacillus cereus and Gram-negative bacteria Escherichia coli, Salmonella Typhimurium and Klebsiella pneumoniae. Fresh leaves of A. indica (neem) was procured from the Department of Botany, JNKVV, Jabalpur. Five samples were taken, and each sample was divided into five subsamples and separated for further isolation of endophytic bacteria. For sterilization leaves were treated with double distilled water, 0.1% sodium hypochlorite, 0.01% bavistin, 0.05% and 70% ethanol. Sterilized leaves of the plants were embedded in Kings B (KB) petri plates and incubated at 37°C for 24 h. Characterization of the bacteria was done according to its morphology and by Gram-staining. After that, a single colony was transferred into brain heart infusion (BHI) broth and incubated at 37°C for 24 h. The antibacterial effect was studied by the disk diffusion method with known antibiotic ciprofloxacin (Ci) as standard. A total of 25 bacterial isolates from A. indica (neem) were obtained and identified morphologically. Most of the samples on KB media depicted irregular shape, flat elevation, undulated, rough, opaque, and white in color. Most of the samples on blood agar showed irregular, raise elevation, undulated, smooth, opaque and all the isolates were nonhemolytic and nonchromogenic. The growth of endophytic bacteria in BHI broth were all isolates showed turbidity. The microscopic examination revealed that maximum isolates were Gram-positive and rod shaped. Good antibacterial activity was observed against S. aureus, Streptococcus pyogenes, E. coli, Salmonella Typhimurium, and K. pneumoniae. Endophytic bacteria are present in leaves of A. indica (neem) and it possesses antibacterial activity against few Gram-positive and Gram

  8. Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves

    PubMed Central

    Singh, Ankit Kumar; Sharma, Rajesh Kumar; Sharma, Varsha; Singh, Tanmay; Kumar, Rajesh; Kumari, Dimple

    2017-01-01

    Aim: The objective of this study was to isolate endophytic bacteria from Azadirachta indica (neem) leaves, their identification and investigate their antibacterial activity against three Gram-positive bacteria, Staphylococcus aureus, Streptococcus pyogenes and Bacillus cereus and Gram-negative bacteria Escherichia coli, Salmonella Typhimurium and Klebsiella pneumoniae. Materials and Methods: Fresh leaves of A. indica (neem) was procured from the Department of Botany, JNKVV, Jabalpur. Five samples were taken, and each sample was divided into five subsamples and separated for further isolation of endophytic bacteria. For sterilization leaves were treated with double distilled water, 0.1% sodium hypochlorite, 0.01% bavistin, 0.05% and 70% ethanol. Sterilized leaves of the plants were embedded in Kings B (KB) petri plates and incubated at 37°C for 24 h. Characterization of the bacteria was done according to its morphology and by Gram-staining. After that, a single colony was transferred into brain heart infusion (BHI) broth and incubated at 37°C for 24 h. The antibacterial effect was studied by the disk diffusion method with known antibiotic ciprofloxacin (Ci) as standard. Results: A total of 25 bacterial isolates from A. indica (neem) were obtained and identified morphologically. Most of the samples on KB media depicted irregular shape, flat elevation, undulated, rough, opaque, and white in color. Most of the samples on blood agar showed irregular, raise elevation, undulated, smooth, opaque and all the isolates were nonhemolytic and nonchromogenic. The growth of endophytic bacteria in BHI broth were all isolates showed turbidity. The microscopic examination revealed that maximum isolates were Gram-positive and rod shaped. Good antibacterial activity was observed against S. aureus, Streptococcus pyogenes, E. coli, Salmonella Typhimurium, and K. pneumoniae. Conclusions: Endophytic bacteria are present in leaves of A. indica (neem) and it possesses antibacterial

  9. Operational taxonomic units (OTUs) of endophytic bacteria isolated from banana cultivars in the Amazon.

    PubMed

    Souza, Á; Nogueira, V B; Cruz, J C; Sousa, N R; Procópio, A R L; Silva, G F

    2015-08-19

    Endophytic microorganisms colonize plants, inhibit the growth of pathogens (by competing for nutrients and/or space), or produce antagonistic substances. Fifty-five endophytic bacteria were isolated from the leaf tissue of the FHIA 18 banana cultivar. Genetic diversity analyses were performed using the enterobacterial repetitive intergenic consensus sequence polymerase chain reaction method and BOX molecular markers. These analyses resulted in 33 and 21 polymorphic bands, respectively. The similarity data, obtained using the Dice coefficient based on the polyphasic analysis method, ranged from 22 to 100%. This indicated a high genetic diversity among the analyzed isolates. Sixty percent similarity was utilized as the cut-off criterion for the formation of operational taxonomic units (OTUs); this resulted in the identification of 32 possible OTUs, indicating a high number of potential species.

  10. Stemphol galactoside, a new stemphol derivative isolated from the tropical endophytic fungus Gaeumannomyces amomi.

    PubMed

    Jumpathong, Juangjun; Abdalla, Muna Ali; Lumyong, Saisamorn; Laatsch, Hartmut

    2010-04-01

    A new stemphol derivative, stemphol 1-O-beta-D-galactopyranoside (1b), together with three known metabolites, stemphol (1a), indole-3-carboxylic acid, and kojic acid, has been isolated from the ethyl acetate extract of cultures of Gaeumannomyces amomi BCC4066, an endophytic fungus found on healthy parts of ginger (Alpinia malaccensis; Thai name Kha-pa). The structure of 1b was established via spectroscopic methods, including 2D NMR measurements and GC/MS experiments.

  11. Broad-Spectrum Antimicrobial Epiphytic and Endophytic Fungi from Marine Organisms: Isolation, Bioassay and Taxonomy

    PubMed Central

    Zhang, Yi; Mu, Jun; Feng, Yan; Kang, Yue; Zhang, Jia; Gu, Peng-Juan; Wang, Yu; Ma, Li-Fang; Zhu, Yan-Hua

    2009-01-01

    In the search for new marine derived antibiotics, 43 epi- and endophytic fungal strains were isolated from the surface or the inner tissue of different marine plants and invertebrates. Through preliminary and secondary screening, 10 of them were found to be able to produce broad-spectrum antimicrobial metabolites. By morphological and molecular biological methods, three active strains were characterized to be Penicillium glabrum, Fusarium oxysporum, and Alternaria alternata. PMID:19597575

  12. Broad-spectrum antimicrobial epiphytic and endophytic fungi from marine organisms: isolation, bioassay and taxonomy.

    PubMed

    Zhang, Yi; Mu, Jun; Feng, Yan; Kang, Yue; Zhang, Jia; Gu, Peng-Juan; Wang, Yu; Ma, Li-Fang; Zhu, Yan-Hua

    2009-04-17

    In the search for new marine derived antibiotics, 43 epi- and endophytic fungal strains were isolated from the surface or the inner tissue of different marine plants and invertebrates. Through preliminary and secondary screening, 10 of them were found to be able to produce broad-spectrum antimicrobial metabolites. By morphological and molecular biological methods, three active strains were characterized to be Penicillium glabrum, Fusarium oxysporum, and Alternaria alternata.

  13. Terpenoids from endophytic fungi.

    PubMed

    de Souza, Jucimar Jorgeane; Vieira, Ivo José Curcino; Rodrigues-Filho, Edson; Braz-Filho, Raimundo

    2011-12-19

    This work reviews the production of terpenoids by endophytic fungi and their biological activities, in period of 2006 to 2010. Sixty five sesquiterpenes, 45 diterpenes, five meroterpenes and 12 other terpenes, amounting to 127 terpenoids were isolated from endophytic fungi.

  14. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    SciTech Connect

    Hirsch, A. M.; Alvarado, J.; Bruce, D.; Chertkov, O.; De Hoff, P. L.; Detter, J. C.; Fujishige, N. A.; Goodwin, L. A.; Han, J.; Han, S.; Ivanova, N.; Land, M. L.; Lum, M. R.; Milani-Nejad, N.; Nolan, M.; Pati, A.; Pitluck, S.; Tran, S. S.; Woyke, T.; Valdes, M.

    2013-08-29

    Micromonospora species live in diverse environments and exhibit a broad range of functions including antibiotic production, biocontrol, and ability to degrade complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico.

  15. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    PubMed Central

    Alvarado, Johana; Bruce, David; Chertkov, Olga; De Hoff, Peter L.; Detter, John C.; Fujishige, Nancy A.; Goodwin, Lynne A.; Han, James; Han, Shunsheng; Ivanova, Natalia; Land, Miriam L.; Lum, Michelle R.; Milani-Nejad, Nima; Nolan, Matt; Pati, Amrita; Pitluck, Sam; Tran, Stephen S.; Woyke, Tanja; Valdés, Maria

    2013-01-01

    Micromonospora species live in diverse environments and exhibit a broad range of functions, including antibiotic production, biocontrol, and degradation of complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico. PMID:24072863

  16. Endophytic fungi isolated from Pinus halepensis needles in M'sila (Algeria) region and their bioactivities.

    PubMed

    Ladjal, S; Harzallah, D; Dahamna, S; Bouamra, D; Bouharati, S; Khennouf, S

    2013-01-01

    Fungal endophytes grow asymptomatically within the tissues of all vascular plants and some are known to provide their host plants with tolerance to different types of environmental stress. Endophytic fungi were detected, isolated and collected from healthy needles of Alep pine (Pinus halepensis Mill.) from the canton of El-Haourane in M'sila, Algeria. For the author's knowledge, these fungi are reported from this conifer for the first time in this region. A survey has been conducted using randomly sampled needles of symptomless trees for two categories of needles (old and young). From 600 surface-sterilized needles collected from 15 trees (300 needles for each age), 29 fungal isolates were obtained and identified on the basis of the morphological characterization using microscopic observations. The fungal isolates were grouped in Deuteromycetes and Zygomycetes. Species of Moniliales were the most common fungi assaciated with P. halepensis. It appears from the results also that the colonization frequency (CF%) was dependant on the age of the needles; older needles were more densely colonized than the younger needles with a 68.5%. The results of this study indicate that P. halepensis serves as a host to numerous endophytic fungi. These fungi could have significance as a source of novel metabolites. And therefore the mycoendophytes genera of Penicillium, Acremonium, Aspergillus, Rhizopus, Trichosporon, Cladosporium, Fusarium, Trichoderma. Expressed an antagonistic and antibiotic effect against pathogenic microorganisms to humans and plants.

  17. Optimization of isolation and cultivation of bacterial endophytes through addition of plant extract to nutrient media.

    PubMed

    Eevers, N; Gielen, M; Sánchez-López, A; Jaspers, S; White, J C; Vangronsveld, J; Weyens, N

    2015-07-01

    Many endophytes have beneficial effects on plants and can be exploited in biotechnological applications. Studies hypothesize that only 0.001-1% of all plant-associated bacteria are cultivable. Moreover, even after successful isolations, many endophytic bacteria often show reduced regrowth capacity. This research aimed to optimize isolation processes and culturing these bacteria afterwards. We compared several minimal and complex media in a screening. Beside the media themselves, two gelling agents and adding plant extract to media were investigated to enhance the number and diversity of endophytes as well as the growth capacity when regrown after isolation. In this work, 869 medium delivered the highest numbers of cultivable bacteria, as well as the highest diversity. When comparing gelling agents, no differences were observed in the numbers of bacteria. Adding plant extract to the media lead to a slight increase in diversity. However, when adding plant extract to improve the regrowth capacity, sharp increases of viable bacteria occurred in both rich and minimal media.

  18. Isolation and Identification of Endophytic Fungi from Actinidia macrosperma and Investigation of Their Bioactivities

    PubMed Central

    Lu, Yin; Chen, Chuan; Chen, Hong; Zhang, Jianfen; Chen, Weiqin

    2012-01-01

    Endophytic fungi from the Chinese medicinal plant Actinidia macrosperma were isolated and identified for the first time. This was the first study to evaluate their cytotoxic and antitumour activities against brine shrimp and five types of tumour cells, respectively. In total, 17 fungal isolates were obtained. Five different taxa were represented by 11 isolates, and six isolates were grouped into the species of Ascomycete Incertae sedis with limited morphological and molecular data. Cytotoxic activity has been found in most isolates except AM05, AM06, and AM10. The isolates AM07 (4.86 μg/mL), AM11 (7.71 μg/mL), and AM17 (14.88 μg/mL) exhibited significant toxicity against brine shrimp. The results of the MTT assay to assess antitumour activity revealed that 82.4% of isolate fermentation broths displayed growth inhibition (50% inhibitory concentration IC50< 100 μg/mL). Moreover, AM07, AM11, and AM17 showed strong antitumour activity in all the cell lines examined. These results suggest that endophytic fungi in A. macrosperma are valuable for the isolation and identification of novel cytotoxic and antitumour bioactive agents. PMID:22203869

  19. [Isolation and identification of root endophytic and rhizosphere bacteria of rice landraces in Yuanyang Terrace, China].

    PubMed

    Zhao, Juan; Liu, Tao; Pan, Lei; Jin, Bai-hui; Zhao, Dan; Chen, Chen; Zhu, You-yong; He, Xia-hong

    2015-12-01

    To reveal root endophytic and rhizosphere bacteria constitution of rice landraces in Yuanyang Terrace, isolation was carried out by tissue isolation method and soil dilution plate method for two landraces of Yuelianggu and Hongjiaolaojing. A total of 399 bacterial strains isolated were identified by morphological characteristics, physiological and biochemical identification. The results showed that there were 8 genera isolated from the root of Yuelianggu and 5 genera from its rhizosphere soil, and 5 genera were same. For Hongjiaolaojing, there were 10 genera isolated from its root and 7 genera from its rhizosphere soil, and 6 genera were same. By molecular biology, identification, a total of 11 species and 5 genera were isolated from the root of Yuelianggu, 8 species and 4 genera from its rhizosphere soil, and 5 species and 4 genera were same. As for Hongjiaolaojing, there were 9 species and 5 genera isolated from its root, and 10 species and 3 genera from its rhizosphere soil, and 4 species and 2 genera were same. The results of physiological and biochemical characteristics identification method and molecular identification method were basically same at the genus level, while most of the strains could be identified to species by molecular identification. There were certain species homology and specificity in the root endophytic and rhizosphere bacteria of Yuanyang rice landraces.

  20. Indoleacetic acid production and plant growth promoting potential of bacterial endophytes isolated from rice (Oryza sativa L.) seeds.

    PubMed

    Shahzad, Raheem; Waqas, Muhammad; Khan, Abdul Latif; Al-Hosni, Khadija; Kang, Sang-Mo; Seo, Chang-Woo; Lee, In-Jung

    2017-06-01

    Bacterial endophytes from the phyllosphere and rhizosphere have been used to produce bioactive metabolites and to promote plant growth. However, little is known about the endophytes residing in seeds. This study aimed to isolate and identify seed-borne bacterial endophytes from rice and elucidate their potential for phytohormone production and growth enhancement. The isolated endophytes included Micrococcus yunnanensis RWL-2, Micrococcus luteus RWL-3, Enterobacter soli RWL-4, Leclercia adecarboxylata RWL-5, Pantoea dispersa RWL-6, and Staphylococcus epidermidis RWL-7, which were identified using 16S rRNA sequencing and phylogenetic analysis. These strains were analyzed for indoleacetic acid (IAA) production by using GC-MS and IAA was found in the range of 11.50 ± 0.77 μg ml(-1) to 38.80 ± 1.35 μg ml(-1). We also assessed the strains for plant growth promoting potential because these isolates were able to produce IAA in pure culture. Most of the growth attributes of rice plants (shoot and root length, fresh and dry biomass, and chlorophyll content) were significantly increased by bacterial endophytes compared to the controls. These results show that IAA producing bacterial endophytes can improve hostplant growth traits and can be used as bio-fertilizers.

  1. Comparative genomics of an endophytic Pseudomonas putida isolated from mango orchard.

    PubMed

    Asif, Huma; Studholme, David J; Khan, Asifullah; Aurongzeb, M; Khan, Ishtiaq A; Azim, M Kamran

    2016-01-01

    We analyzed the genome sequence of an endophytic bacterial strain Pseudomonas putida TJI51 isolated from mango bark tissues. Next generation DNA sequencing and short read de novo assembly generated the 5,805,096 bp draft genome of P. putida TJI51. Out of 6,036 protein coding genes in P. putida TJI51 sequences, 4,367 (72%) were annotated with functional specifications, while the remaining encoded hypothetical proteins. Comparative genome sequence analysis revealed that the P. putida TJI51genome contains several regions, not identified in so far sequenced P. putida genomes. Some of these regions were predicted to encode enzymes, including acetylornithine deacetylase, betaine aldehyde dehydrogenase, aldehyde dehydrogenase, benzoylformate decarboxylase, hydroxyacylglutathione hydrolase, and uroporphyrinogen decarboxylase. The genome of P. putida TJI51 contained three nonribosomal peptide synthetase gene clusters. Genome sequence analysis of P. putidaTJI51 identified this bacterium as an endophytic resident. The endophytic fitness might be linked with alginate, which facilitates bacterial colonization in plant tissues. Genome sequence analysis shed light on the presence of a diverse spectrum of metabolic activities and adaptation of this isolate to various niches.

  2. Comparative genomics of an endophytic Pseudomonas putida isolated from mango orchard

    PubMed Central

    Asif, Huma; Studholme, David J.; Khan, Asifullah; Aurongzeb, M.; Khan, Ishtiaq A.; Azim, M. Kamran

    2016-01-01

    Abstract We analyzed the genome sequence of an endophytic bacterial strain Pseudomonas putida TJI51 isolated from mango bark tissues. Next generation DNA sequencing and short read de novo assembly generated the 5,805,096 bp draft genome of P. putida TJI51. Out of 6,036 protein coding genes in P. putida TJI51 sequences, 4,367 (72%) were annotated with functional specifications, while the remaining encoded hypothetical proteins. Comparative genome sequence analysis revealed that the P. putida TJI51genome contains several regions, not identified in so far sequenced P. putida genomes. Some of these regions were predicted to encode enzymes, including acetylornithine deacetylase, betaine aldehyde dehydrogenase, aldehyde dehydrogenase, benzoylformate decarboxylase, hydroxyacylglutathione hydrolase, and uroporphyrinogen decarboxylase. The genome of P. putida TJI51 contained three nonribosomal peptide synthetase gene clusters. Genome sequence analysis of P. putidaTJI51 identified this bacterium as an endophytic resident. The endophytic fitness might be linked with alginate, which facilitates bacterial colonization in plant tissues. Genome sequence analysis shed light on the presence of a diverse spectrum of metabolic activities and adaptation of this isolate to various niches. PMID:27560648

  3. Characterization of pectin lyase produced by an endophytic strain isolated from coffee cherries.

    PubMed

    Sakiyama, C C; Paula, E M; Pereira, P C; Borges, A C; Silva, D O

    2001-08-01

    The effect of endophytic bacterial activity on the quality of coffee beverage was studied. A survey of the micro-organisms in coffee cherries was performed before harvesting, and their growth on the main nutrients available in coffee cherries was determined in vitro. Many endophytic bacteria were isolated from surface-sterilized coffee cherries. One of the pectinolytic strains was physiologically and phenotypically characterized, and was tentatively identified by partial 16S rDNA sequencing as Paenibacillus amylolyticus. This endophytic strain produced an extracellular pectinase with maximal activity at 40 degrees C and pH 7.9, and was thermostable up to 45 degrees C. EDTA and metal ions had little effect on pectin lyase activity. Km and Vmax values were 4.6 mg ml(-1) and 94.0 10(-8) mol min(-1) ml(-1), respectively. Pectin lyases have been found in fungi but rarely in bacteria, and this isolate is a promising tool for regulation studies of these enzymes.

  4. Isolation and characterization of culturable seed-associated bacterial endophytes from gnotobiotically grown Marama bean seedlings.

    PubMed

    Chimwamurombe, Percy Maruwa; Grönemeyer, Jann Lasse; Reinhold-Hurek, Barbara

    2016-06-01

    Marama bean (Tylosema esculentum) is an indigenous non-nodulating legume to the arid agro-ecological parts of Southern Africa. It is a staple food for the Khoisan and Bantu people from these areas. It is intriguing how it is able to synthesize the high-protein content in the seeds since its natural habitat is nitrogen deficient. The aim of the study was to determine the presence of seed transmittable bacterial endophytes that may have growth promoting effects, which may be particularly important for the harsh conditions. Marama bean seeds were surface sterilized and gnotobiotically grown to 2 weeks old seedlings. From surface-sterilized shoots and roots, 123 distinct bacterial isolates were cultured using three media, and identified by BOX-PCR fingerprinting and sequence analyses of the 16S rRNA and nifH genes. Phylogenetic analyses of 73 putative endophytes assigned them to bacterial species from 14 genera including Proteobacteria (Rhizobium, Massilia, Kosakonia, Pseudorhodoferax, Caulobacter, Pantoea, Sphingomonas, Burkholderia, Methylobacterium), Firmicutes (Bacillus), Actinobacteria (Curtobacterium, Microbacterium) and Bacteroidetes (Mucilaginibacter, Chitinophaga). Screening for plant growth-promoting activities revealed that the isolates showed production of IAA, ACC deaminase, siderophores, endoglucanase, protease, AHLs and capacities to solubilize phosphate and fix nitrogen. This is the first report that marama bean seeds may harbor endophytes that can be cultivated from seedlings; in this community of bacteria, physiological characteristics that are potentially plant growth promoting are widespread.

  5. Dietzia papillomatosis sp. nov., a novel actinomycete isolated from the skin of an immunocompetent patient with confluent and reticulated papillomatosis.

    PubMed

    Jones, Amanda L; Koerner, Roland J; Natarajan, Sivakumar; Perry, John D; Goodfellow, Michael

    2008-01-01

    An actinomycete isolated from an immunocompetent patient suffering from confluent and reticulated papillomatosis was characterized using a polyphasic taxonomic approach. The organism had chemotaxonomic and morphological properties that were consistent with its assignment to the genus Dietzia and it formed a distinct phyletic line within the Dietzia 16S rRNA gene tree. It shared a 16S rRNA gene sequence similarity of 98.3% with its nearest neighbour, the type strain of Dietzia cinnamea, and could be distinguished from the type strains of all Dietzia species using a combination of phenotypic properties. It is apparent from genotypic and phenotypic data that the organism represents a novel species in the genus Dietzia. The name proposed for this taxon is Dietzia papillomatosis; the type strain is N 1280(T) (=DSM 44961(T)=NCIMB 14145(T)).

  6. Richness of endophytic fungi isolated from Opuntia ficus-indica Mill. (Cactaceae) and preliminary screening for enzyme production.

    PubMed

    Bezerra, J D P; Santos, M G S; Svedese, V M; Lima, D M M; Fernandes, M J S; Paiva, L M; Souza-Motta, C M

    2012-05-01

    Opuntia ficus-indica Mill. (forage cactus) is farmed with relative success in the semi-arid region of the Brazilian northeast for commercial purposes, particularly as forage and food. Endophytic microorganisms are those that can be isolated inside plant tissues and can be a new source to production of enzymes with different potentialities. The objective of this study was to describe the richness of endophytic fungi from O. ficus-indica and to detect the capacity of these species to produce extracellular hydrolytic enzymes. Forty-four endophytic fungi species were isolated. Among them, the most commonly found were Cladosporium cladosporioides (20.43%) and C. sphaerospermum (15.99%). Acremonium terricola, Monodictys castaneae, Penicillium glandicola, Phoma tropica and Tetraploa aristata are being reported for the first time as endophytic fungi for Brazil. The majority of isolated fungi exhibited enzymatic potential. Aspergillus japonicus and P. glandicola presented pectinolytic activity. Xylaria sp. was the most important among the other 14 species with positive cellulase activity. All 24 isolates analysed were xylanase-positive. Protease was best produced by isolate PF103. The results indicate that there is a significant richness of endophytic fungi in O. ficus-indica, and that these isolates indicate promising potential for deployment in biotechnological processes involving production of pectinases, cellulases, xylanases and proteases.

  7. [Stress resistance and genetic diversity of endophytic bacteria isolated from Caragana spp. root nodules].

    PubMed

    Dai, Jin-Xia; Wang, Yu-Jiong; Wu, Xue-Juan; Zhang, Xiao-Ling

    2012-02-01

    By adopting PCR-RFLP and 16S rDNA sequencing, this paper analyzed the genetic diversity and phylogeny of 40 endophytic bacterial strains isolated from Caragana spp. root nodules, and determined the salt resistance, acid- and alkali resistance, and growth temperature range of the strains. A total of 9 genotypes were obtained from the 40 strains by RFLP. The 16S rDNA sequencing, morphological observation, and biochemical test of representative strains showed that the strains belonged to Bacillus, Inquilinus, Shinella and Acinetobacter, respectively, and had rich genetic diversity. 57.5% of the strains could tolerate 4% NaCl stress, 75% of the strains could grow in YMA medium with an initial pH 11.0, and 85% of the strains could survive after heat shock treatment at 60 degrees C, suggesting that the endophytic bacteria of Caragana spp. had strong resistance capacity. Among the strains, LWEN 07 and LWEN 15 were most resistant.

  8. Lovastatin-producing endophytic fungus isolated from a medicinal plant Solanum xanthocarpum.

    PubMed

    Parthasarathy, Ramalingam; Sathiyabama, Muthukrishnan

    2015-01-01

    Lovastatin is a potent drug for lowering blood cholesterol. An endophytic fungus Phomopsis vexans was isolated from the healthy leaf tissues of Solanum xanthocarpum, a medicinal plant, and screened for lovastatin production. The fungus was identified by their characteristic cultural morphology and molecular analysis. The strain had a component with the same TLC Rf value and HPLC retention time as authentic lovastatin. The presence of lovastatin was further confirmed by FT-IR, UV, (1)H, (13)C NMR and LC-MS analyses. The amount of lovastatin produced by this endophytic fungus was quantified to be 550 mg/L, and thus the fungus can serve as a potential material to improve the production of lovastatin.

  9. Discovery of novel metabolites from marine actinomycetes.

    PubMed

    Lam, Kin S

    2006-06-01

    Recent findings from culture-dependent and culture-independent methods have demonstrated that indigenous marine actinomycetes exist in the oceans and are widely distributed in different marine ecosystems. There is tremendous diversity and novelty among the marine actinomycetes present in marine environments. Progress has been made to isolate novel actinomycetes from samples collected at different marine environments and habitats. These marine actinomycetes produce different types of new secondary metabolites. Many of these metabolites possess biological activities and have the potential to be developed as therapeutic agents. Marine actinomycetes are a prolific but underexploited source for the discovery of novel secondary metabolites.

  10. Biocontrol of verticillium wilt and colonization of cotton plants by an endophytic bacterial isolate.

    PubMed

    Li, C-H; Shi, L; Han, Q; Hu, H-L; Zhao, M-W; Tang, C-M; Li, S-P

    2012-09-01

    To explore biocontrol potential of 39 DAEB isolates (doubly antagonistic towards both Verticillium dahliae Kleb and Fusarium oxysporum) against verticillium wilt of cotton and to elucidate colonization and category characteristics of an endophytic bacterium with significant biocontrol activity. Thirty-nine antagonistic endophytic bacteria strains were tested for their ability to control verticillium wilt in cotton plants caused by a defoliating pathotype of V. dahliae 107 in cotton under controlled conditions. The biocontrol trial revealed that an endophytic bacterium, designated HA02, showed a significant biocontrol activity to V. dahliae 107. After cotton seedlings were inoculated with a gfp gene-tagged HA02 (HA02-gfp), HA02-gfp populations were higher in the root than in the stem; in addition, the HA02-gfp was distributed in the maturation zone of cotton root. Furthermore, HA02-gfp also colonized seedlings of maize, rape and soybean after the bacteria inoculation. Phylogenetic trees based on 16S rDNA sequences combined with morphological, physiological and identification showed that the bacterium belongs to the Enterobacter genus. Our results showed that only 1 of 39 DAEB isolates demonstrated more efficient biocontrol potential towards V. dahliae 107 in greenhouse and field trials. HA02-gfp mainly colonized cotton in roots. In addition, we quantitatively observed HA02 colonization in other hosts. HA02 belongs to the Enterobacter genus. This is the first study on biocontrol to defoliating pathotype of V. dahliae Kleb by endophytic bacteria. The HA02 showed effective biocontrol to V. dahliae 107 in greenhouse and field trials. Furthermore, we assessed the quantitative and qualitative colonization of HA02 in cotton seedlings. Our study provides basic information to further explore managing strategies to control this critical disease. © 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  11. Anti-Respiratory Syncytial Virus Compounds from Two Endophytic Fungi Isolated from Nigerian Medicinal Plants.

    PubMed

    Uzor, P F; Odimegwu, D C; Ebrahim, W; Osadebe, P O; Nwodo, N J; Okoye, F B C; Liu, Z; Proksch, P

    2016-10-01

    Background: Respiratory syncytial virus (RSV) is known to cause severe respiratory infections particularly in infants younger than 2 years of age. The only approved drug, ribavirin, is expensive and is not likely to improve therapeutic outcome, thereby necessitating the search for safer and more potent alternatives from natural sources such as endophytic fungi. The present study aimed to investigate the anti-RSV activity of compounds from endophytic fungi. Methods: Two endophytic fungi Colletotrichum gloeosporioides and Pestalotiopsis thea were isolated from the fresh leaves of the host Nigerian plants Anthocleista djalonensis and Fagara zanthoxyloides, respectively. After fermentation in solid rice media, C. gloeosporioides afforded 4 known compounds 4-hydroxybenzoic acid (1), vanillic acid (2), ferulic acid (3) and Nb-acetyltryptamine (4) while P. thea afforded 3 known compounds chloroisosulochrin (5), ficipyrone A (6) and pestheic acid (7). The compounds were investigated for their anti-RSV activity using the HEP-2 cell lines and ribavirin as the standard drug. Results: Compound 5 was found to show the strongest inhibition of the RSV with IC50 of 4.22±1.03 µM (ribavirin 4.91±1.85 µM). Other compounds showed moderate inhibition of the virus (IC50 ranging from 45.00±0.98 to 259.23±2.36 µM). Conclusion: The results of the present study have shown that chloroisosulochrin (5), isolated from an endophytic fungus P. thea, possesses strong activity against RSV. © Georg Thieme Verlag KG Stuttgart · New York.

  12. Plantactinospora endophytica sp. nov., an actinomycete isolated from Camptotheca acuminata Decne., reclassification of Actinaurispora siamensis as Plantactinospora siamensis comb. nov. and emended descriptions of the genus Plantactinospora and Plantactinospora mayteni.

    PubMed

    Zhu, Wen-Yong; Zhao, Li-Xing; Zhao, Guo-Zhen; Duan, Xue-Wei; Qin, Sheng; Li, Jie; Xu, Li-Hua; Li, Wen-Jun

    2012-10-01

    A novel endophytic actinomycete, designated strain YIM 68255(T), was isolated from healthy leaves of Camptotheca acuminata Decne. collected in Yunnan province, south-west China and characterized by using a polyphasic approach. The strain formed well-developed substrate mycelium, but no aerial mycelium. It grew at 10-45 °C, at pH 5-10 (optimum pH 7) and in the presence of 0-3 % (w/v) NaCl. The DNA G+C content was 73.0 mol%. Phylogenetic analyses showed that strain YIM 68255(T) belonged to the genus Plantactinospora. However, it exhibited some differences from Plantactinospora mayteni YIM 61359(T) and the level of DNA-DNA relatedness was 42.7 ± 1.3 %. Based on comparative analysis of physiological and chemotaxonomic data, it is proposed that strain YIM 68255(T) represents a novel species of the genus Plantactinospora, Plantactinospora endophytica sp. nov., with strain YIM 68255(T) ( = DSM 45387(T) = CCTCC AA 209047(T)) as the type strain. In addition, it is also proposed that Actinaurispora siamensis Thawai et al. 2010 be transferred to the genus Plantactinospora as Plantactinospora siamensis comb. nov. [type strain CM2-8(T) ( = JCM 15677(T) = BCC 34762(T))] based on chemotaxonomic characteristics and phylogenetic analysis. Emended descriptions of the genus Plantactinospora and Plantactinospora mayteni are also provided.

  13. In vitro and in vivo antagonism of actinomycetes isolated from Moroccan rhizospherical soils against Sclerotium rolfsii: a causal agent of root rot on sugar beet (Beta vulgaris L.).

    PubMed

    Errakhi, R; Lebrihi, A; Barakate, M

    2009-08-01

    To evaluate the ability of the isolated actinomycetes to inhibit in vitro plant pathogenic fungi and the efficacy of promising antagonistic isolates to reduce in vivo the incidence of root rot induced by Sclerotium rolfsii on sugar beet. Actinomycetes isolated from rhizosphere soil of sugar beet were screened for antagonistic activity against a number of plant pathogens, including S. rolfsii. Ten actinomycetes out of 195 screened in vitro were strongly inhibitory to S. rolfsii. These isolates were subsequently tested for their ability to inhibit sclerotial germination and hyphal growth of S. roflsii. The most important inhibitions were obtained by the culture filtrate from the isolates J-2 and B-11, including 100% inhibition of sclerotial germination and 80% inhibition of hyphal growth. These two isolates (J-2 and B-11) were then screened for their ability to protect sugar beet against infection of S. rolfsii induced root rot in a pot trial. The treatment of S. rolfsii infested soil with a biomass and culture filtrate mixture of the selected antagonists reduced significantly (P < or = 0.05) the incidence of root rot on sugar beet. Isolate J-2 was most effective and allowed a high fresh weight of sugar beet roots to be obtained. Both antagonists J-2 and B-11 were classified as belonging to the genus Streptomyces species through morphological and chemical characteristics as well as 16S rDNA analysis. Streptomyces isolates J-2 and B-11 showed a potential for controlling root rot on sugar beet and could be useful in integrated control against diverse soil borne plant pathogens. This investigation showed the role, which actinomycete bacteria can play to control root rot caused by S. rolfsii, in the objective to reduce treatments with chemical fungicides.

  14. Diversity and antimicrobial activity of endophytic fungi isolated from Cephalotaxus hainanensis Li, a well-known medicinal plant in China.

    PubMed

    Yang, H R; Hu, X P; Jiang, C J; Qi, J; Wu, Y C; Li, W; Zeng, Y J; Li, C F; Liu, S X

    2015-11-01

    About 1051 endophytic fungi were isolated from leaves, branches, barks and stems of Cephalotaxus hainanensis Li from four sites in Hainan, China. The fungi were identified as 21 genera by morphology and ITS sequences. One dominant species was Phomopsis quercella in Hainan Tropical Botanical Garden and Bawangling Nature Reserve, with relative frequency of 42·06 and 34·88% respectively. Another dominant species was Colletotrichum boninense in Wuzhishan and Jianfengling Nature Reserves, with relative frequency of 36·84 and 46·97% respectively. Among the selected 21 endophytic fungi, 17 strains (80·95%) had activity against at least one pathogenic bacteria, and 14 strains (66·67%) exhibited activity against at least one fungal pathogens. Neonectria macroconidialis showed strong inhibition against Staphylococcus aureus (inhibition zone being 20 mm), Bacillus subtilis (14 mm) and Streptococcus agalactiae (28 mm). Xylaria sp. showed strong inhibition against Escherichia coli (20 mm), Rhizoctonia solani (20 mm) and Sclerotinia sclerotiorum (17 mm). Verticillium bulbillosum showed great activity against Strep. agalactiae (32 mm) and Fusarium oxysporum (22 mm). These endophytic fungi showed potentials in medicine development. Endophytic fungi from medicinal plants are an important source of novel and viable drugs. Cephalotaxus hainanensis Li is well known for leukaemia treatment and its endophytic fungi were isolated to investigate the diversity and antimicrobial activity. It was found that Ce. hainanensis Li had rich endophytic fungi, and some fungi showed strong antimicrobial activity against certain pathogens. These fungi can be used in medicine development. © 2015 The Society for Applied Microbiology.

  15. Isolation, diversity and acetylcholinesterase inhibitory activity of the culturable endophytic fungi harboured in Huperzia serrata from Jinggang Mountain, China.

    PubMed

    Wang, Ya; Lai, Zheng; Li, Xi-Xi; Yan, Ri-Ming; Zhang, Zhi-Bin; Yang, Hui-Lin; Zhu, Du

    2016-02-01

    Huperzia serrata has many important medicinal properties with proven pharmacological potential. Some of these properties may be mediated by its endophytic fungi. To test this hypothesis, in the present study, we provided a first insights into evaluating the species composition and acetylcholinesterase (AChE) inhibitory activity of the culturable endophytic fungi of H. serrata from the regional at Jinggang Mountain in southeastern China. A total number of 885 fungal isolates distributed across 44 genera and 118 putative species were obtained from 1422 fragments of fine H. serrata roots, stems and leaves base on ITS-rDNA sequences BLAST analysis. The endophytic fungi were phylogenetically diverse and species-rich, with high rate of colonization and isolation. The assemble of endophytic fungi consisted mainly of Ascomycota (97.15%), followed by Basidiomycota (1.92%) and unknown fungal species (0.90%). Colletotrichum (64.29%), Phyllosticta (3.39%), Hypoxylon (2.81%), Xylaria (2.25%) and Nigrospora (2.04%) were the most abundant genera, whereas the remaining genera were infrequent groups. Although, roots yielded low abundance strains, the diverse and species-rich were both higher than that of stems and leaves. In addition, out of the 247 endophytic fungi strains determinated, 221 fungal extracts showed AChE inhibition activities in vitro. Among them, 22 endophytic fungi strains achieved high inhibitory activity (≥50%) on AChE which belongs to 13 genera and five incertae sedis strains. Four endophytic fungi designated as JS4 (Colletotrichum spp.), FL14 (Ascomycota spp.), FL9 (Sarcosomataceae spp.) and FL7 (Dothideomycetes spp.) were displayed highly active (≥80%) against AChE, which the inhibition effects were even more intense than the positive control. Our findings highlight that H. serrata grown in Jinggang Mountain harbors a rich and fascinating endophytic fungus community with potential AChE inhibitory activity, which could further broaden the natural

  16. Plant growth-promoting potential of endophytic bacteria isolated from roots of coastal sand dune plants.

    PubMed

    Shin, Dong-Sung; Park, Myung Soo; Jung, Sera; Lee, Myoung Sook; Lee, Kang Hyun; Bae, Kyung Sook; Kim, Seung Bum

    2007-08-01

    Endophytic bacteria associated with the roots of coastal sand dune plants were isolated, taxonomically characterized, and tested for their plant growth-promoting activities. Ninety-one endophytic bacterial isolates were collected and assigned to 17 different genera of 6 major bacterial phyla based on partial 16S rDNA sequence analyses. Gammaproteobacteria represented the majority of the isolates (65.9%), and members of Pseudomonas constituted 49.5% of the total isolates. When testing for antagonism towards plant pathogenic fungi, 25 strains were antagonistic towards Rhizoctonia solani, 57 strains were antagonistic towards Pythium ultimum, 53 strains were antagonistic towards Fusarium oxysporum, and 41 strains were antagonistic towards Botrytis cinerea. Seven strains were shown to produce indole acetic acid (IAA), 33 to produce siderophores, 23 to produce protease, 37 to produce pectinase, and 38 to produce chitinase. The broadest spectra of activities were observed among the Pseudomonas strains, indicating outstanding plant growth-promoting potential. The isolates from C. kobomugi and M. sibirica also exhibited good plant growth-promoting potential. The correlations among individual plant growth-promoting activities were examined using phi coefficients, and the resulting data indicated that the production of protease, pectinase, chitinase, and siderophores was highly related.

  17. Endophytic Actinobacteria Associated with Dracaena cochinchinensis Lour.: Isolation, Diversity, and Their Cytotoxic Activities.

    PubMed

    Salam, Nimaichand; Khieu, Thi-Nhan; Liu, Min-Jiao; Vu, Thu-Trang; Chu-Ky, Son; Quach, Ngoc-Tung; Phi, Quyet-Tien; Narsing Rao, Manik Prabhu; Fontana, Angélique; Sarter, Samira; Li, Wen-Jun

    2017-01-01

    Dracaena cochinchinensis Lour. is an ethnomedicinally important plant used in traditional Chinese medicine known as dragon's blood. Excessive utilization of the plant for extraction of dragon's blood had resulted in the destruction of the important niche. During a study to provide a sustainable way of utilizing the resources, the endophytic Actinobacteria associated with the plant were explored for potential utilization of their medicinal properties. Three hundred and four endophytic Actinobacteria belonging to the genera Streptomyces, Nocardiopsis, Brevibacterium, Microbacterium, Tsukamurella, Arthrobacter, Brachybacterium, Nocardia, Rhodococcus, Kocuria, Nocardioides, and Pseudonocardia were isolated from different tissues of D. cochinchinensis Lour. Of these, 17 strains having antimicrobial and anthracyclines-producing activities were further selected for screening of antifungal and cytotoxic activities against two human cancer cell lines, MCF-7 and Hep G2. Ten of these selected endophytic Actinobacteria showed antifungal activities against at least one of the fungal pathogens, of which three strains exhibited cytotoxic activities with IC50-values ranging between 3 and 33 μg·mL(-1). Frequencies for the presence of biosynthetic genes, polyketide synthase- (PKS-) I, PKS-II, and nonribosomal peptide synthetase (NRPS) among these 17 selected bioactive Actinobacteria were 29.4%, 70.6%, and 23.5%, respectively. The results indicated that the medicinal plant D. cochinchinensis Lour. is a good niche of biologically important metabolites-producing Actinobacteria.

  18. Endophytic Actinobacteria Associated with Dracaena cochinchinensis Lour.: Isolation, Diversity, and Their Cytotoxic Activities

    PubMed Central

    Salam, Nimaichand; Khieu, Thi-Nhan; Liu, Min-Jiao; Vu, Thu-Trang; Quach, Ngoc-Tung; Phi, Quyet-Tien; Fontana, Angélique; Sarter, Samira

    2017-01-01

    Dracaena cochinchinensis Lour. is an ethnomedicinally important plant used in traditional Chinese medicine known as dragon's blood. Excessive utilization of the plant for extraction of dragon's blood had resulted in the destruction of the important niche. During a study to provide a sustainable way of utilizing the resources, the endophytic Actinobacteria associated with the plant were explored for potential utilization of their medicinal properties. Three hundred and four endophytic Actinobacteria belonging to the genera Streptomyces, Nocardiopsis, Brevibacterium, Microbacterium, Tsukamurella, Arthrobacter, Brachybacterium, Nocardia, Rhodococcus, Kocuria, Nocardioides, and Pseudonocardia were isolated from different tissues of D. cochinchinensis Lour. Of these, 17 strains having antimicrobial and anthracyclines-producing activities were further selected for screening of antifungal and cytotoxic activities against two human cancer cell lines, MCF-7 and Hep G2. Ten of these selected endophytic Actinobacteria showed antifungal activities against at least one of the fungal pathogens, of which three strains exhibited cytotoxic activities with IC50-values ranging between 3 and 33 μg·mL−1. Frequencies for the presence of biosynthetic genes, polyketide synthase- (PKS-) I, PKS-II, and nonribosomal peptide synthetase (NRPS) among these 17 selected bioactive Actinobacteria were 29.4%, 70.6%, and 23.5%, respectively. The results indicated that the medicinal plant D. cochinchinensis Lour. is a good niche of biologically important metabolites-producing Actinobacteria. PMID:28484706

  19. Taxol production by an endophytic fungus, Fusarium redolens, isolated from Himalayan yew.

    PubMed

    Garyali, Sanjog; Kumar, Anil; Reddy, M Sudhakara

    2013-10-28

    Different endophytic fungi isolated from Himalayan Yew plants were tested for their ability to produce taxol. The BAPT gene (C-13 phenylpropanoid side chain-CoA acetyl transferase) involved in the taxol biosynthetic pathway was used as a molecular marker to screen taxol-producing endophytic fungi. Taxol extracted from fungal strain TBPJ-B was identified by HPLC and MS analysis. Strain TBPJ-B was identified as Fusarium redolens based on the morphology and internal transcribed spacer region of nrDNA analysis. HPLC quantification of fungal taxol showed that F. redolens was capable of producing 66 μg/l of taxol in fermentation broth. The antitumour activity of the fungal taxol was tested by potato disc tumor induction assay using Agrobacterium tumefaciens as the tumor induction agent. The present study results showed that PCR amplification of genes involved in taxol biosynthesis is an efficient and reliable method for prescreening taxol-producing fungi. We are reporting for the first time the production of taxol by F. redolens from Taxus baccata L. subsp. wallichiana (Zucc.) Pilger. This study offers important information and a new source for the production of the important anticancer drug taxol by endophytic fungus fermentation.

  20. Diversity and Antimicrobial Activity of Culturable Endophytic Fungi Isolated from Moso Bamboo Seeds

    PubMed Central

    Cai, Chun-Ju; Fan, Li; Gao, Jian; Hou, Cheng-Lin

    2014-01-01

    Bamboos, regarded as therapeutic agents in ethnomedicine, have been used to inhibit inflammation and enhance natural immunity for a long time in Asia, and there are many bamboo associated fungi with medical and edible value. In the present study, a total of 350 fungal strains were isolated from the uncommon moso bamboo (Phyllostachys edulis) seeds for the first time. The molecular diversity of these endophytic fungi was investigated and bioactive compound producers were screened for the first time. All the fungal endophytes were categorized into 69 morphotypes according to culturable characteristics and their internal transcriber spacer (ITS) regions were analyzed by BLAST search with the NCBI database. The fungal isolates showed high diversity and were divided in Ascomycota (98.0%) and Basidiomycota (2.0%), including at least 19 genera in nine orders. Four particular genera were considered to be newly recorded bambusicolous fungi, including Leptosphaerulina, Simplicillium, Sebacina and an unknown genus in Basidiomycetes. Furthermore, inhibitory effects against clinical pathogens and phytopathogens were screened preliminarily and strains B09 (Cladosporium sp.), B34 (Curvularia sp.), B35 (undefined genus 1), B38 (Penicillium sp.) and zzz816 (Shiraia sp.) displayed broad-spectrum activity against clinical bacteria and yeasts by the agar diffusion method. The crude extracts of isolates B09, B34, B35, B38 and zzz816 under submerged fermentation, also demonstrated various levels of bioactivities against bambusicolous pathogenic fungi. This study is the first report on the antimicrobial activity of endophytic fungi associated with moso bamboo seeds, and the results show that they could be exploited as a potential source of bioactive compounds and plant defense activators. In addition, it is the first time that strains of Shiraia sp. have been isolated and cultured from moso bamboo seeds, and one of them (zzz816) could produce hypocrellin A at high yield, which is

  1. Screening of endophytic fungi having ability for antioxidative and alpha-glucosidase inhibitor activities isolated from Taxus sumatrana.

    PubMed

    Artanti, N; Tachibana, S; Kardono, L B S; Sukiman, H

    2011-11-15

    Endophytic microbes are considered as an important source of natural products. They show antibiotic, anticancer, antioxidative and antidiabetic activities. Therefore, there are many reports on the isolation and bioactivity screening of endophytic fungi from various plants including Taxus species. Taxus sumatrana (Miq.) de Laub is found in Indonesia. The objective of this study is to conduct an in vitro screening of 14 endophytic fungi isolated from Taxus sumatrana having antioxidative and alpha-glucosidase inhibitor activities. Each endophytic fungus was cultured for 7 days and the fungal mycelium and medium were extracted with methanol and ethyl acetate, respectively, to produce each extract. The antioxidative activity of each extract was tested by DPPH free radical scavenging activity and beta-carotene bleaching assays, whereas antidiabetic activity was tested based on alpha-glucosidase inhibitor activity. The screening results showed that fungal mycelia of TSC 13 had the best alpha-glucosidase inhibitor activity and TSC 24 had the best antioxidative activity. Isolation of bioactive compounds from TSC 13 and TSC 24 is being conducted. This is the first report that endophytic fungi isolated from T. sumatrana exhibited anti alpha-glucosidase inhibitory and anti oxidative activities.

  2. Paenibacillus herberti sp. nov., an endophyte isolated from Herbertus sendtneri.

    PubMed

    Guo, Guan Nan; Zhou, Xun; Zhao, Ran; Chen, Xin Yao; Chen, Zhi Ling; Li, Xue Dong; Li, Yan Hong

    2015-09-01

    Strain R33(T), an endophyte recovered from Herbertus sendtneri, was identified as representing a novel species of the genus Paenibacillus by using a polyphasic taxonomic approach. The novel strain was observed to be a Gram-stain positive, aerobic, rod-shaped, motile and endospore-forming bacterium. The major polar lipids of strain R33(T) were identified as diphosphatidylglycerol, phosphatidylethanolamine, along with lesser amounts of phosphatidylglycerol, three unidentified aminophospholipids, two unidentified phospholipids and two unidentified lipids. The predominant isoprenoid quinone was identified as MK-7. The major fatty acids (>8.0 %) were found to be anteiso-C15:0 (40.0 %), C16:1 ω11c (9.4 %), C16:1 ω7c alcohol (8.5 %) and C16:0 (8.2 %). The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The G+C content of genomic DNA was determined to be 56.9 mol%. The 16S rRNA gene sequence similarities of strain R33(T) to other Paenibacillus species ranged from 91.6 to 97.2 %, with high similarities to Paenibacillus humicus PC-147(T) and Paenibacillus pasadenensis SAFN-007(T). The phylogenetic analyses based on 16S rRNA gene sequences and the partial rpoB gene confirmed that strain R33(T) belongs to the genus Paenibacillus. However, strain R33(T) shows differential molecular characteristics compared to other related Paenibacillus species based on 16S rDNA-RFLP analyses; the DNA-DNA relatedness values between strain R33(T) and P. humicus PC-147(T), and that between strain R33(T) and P. pasadenensis SAFN-007(T), were 35.0 ± 2.0 and 41.4 ± 0.9 %, respectively. Based on its phenotypic, chemotaxonomic and phylogenetic properties, strain R33(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus herberti is proposed (type strain R33(T) = CGMCC 1.15042(T) = DSM 29849(T)).

  3. Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars.

    PubMed

    Ji, Sang Hye; Gururani, Mayank Anand; Chun, Se-Chul

    2014-01-20

    We have isolated 576 endophytic bacteria from the leaves, stems, and roots of 10 rice cultivars and identified 12 of them as diazotrophic bacteria using a specific primer set of nif gene. Through 16S rDNA sequence analysis, nifH genes were confirmed in the two species of Penibacillus, three species of Microbacterium, three Bacillus species, and four species of Klebsiella. Rice seeds treated with these plant growth-promoting bacteria (PGPB) showed improved plant growth, increased height and dry weight and antagonistic effects against fungal pathogens. In addition, auxin and siderophore producing ability, and phosphate solubilizing activity were studied for the possible mechanisms of plant growth promotion. Among 12 isolates tested, 10 strains have shown higher auxin producing activity, 6 isolates were confirmed as strains with high siderophore producing activity while 4 isolates turned out to have high phosphate-solubilizing activity. These results strongly suggest that the endophytic diazotrophic bacteria characterized in this study could be successfully used to promote plant growth and inducing fungal resistance in plants.

  4. Endophytic Colonization and In Planta Nitrogen Fixation by a Herbaspirillum sp. Isolated from Wild Rice Species

    PubMed Central

    Elbeltagy, Adel; Nishioka, Kiyo; Sato, Tadashi; Suzuki, Hisa; Ye, Bin; Hamada, Toru; Isawa, Tsuyoshi; Mitsui, Hisayuki; Minamisawa, Kiwamu

    2001-01-01

    Nitrogen-fixing bacteria were isolated from the stems of wild and cultivated rice on a modified Rennie medium. Based on 16S ribosomal DNA (rDNA) sequences, the diazotrophic isolates were phylogenetically close to four genera: Herbaspirillum, Ideonella, Enterobacter, and Azospirillum. Phenotypic properties and signature sequences of 16S rDNA indicated that three isolates (B65, B501, and B512) belong to the Herbaspirillum genus. To examine whether Herbaspirillum sp. strain B501 isolated from wild rice, Oryza officinalis, endophytically colonizes rice plants, the gfp gene encoding green fluorescent protein (GFP) was introduced into the bacteria. Observations by fluorescence stereomicroscopy showed that the GFP-tagged bacteria colonized shoots and seeds of aseptically grown seedlings of the original wild rice after inoculation of the seeds. Conversely, for cultivated rice Oryza sativa, no GFP fluorescence was observed for shoots and only weak signals were observed for seeds. Observations by fluorescence and electron microscopy revealed that Herbaspirillum sp. strain B501 colonized mainly intercellular spaces in the leaves of wild rice. Colony counts of surface-sterilized rice seedlings inoculated with the GFP-tagged bacteria indicated significantly more bacterial populations inside the original wild rice than in cultivated rice varieties. Moreover, after bacterial inoculation, in planta nitrogen fixation in young seedlings of wild rice, O. officinalis, was detected by the acetylene reduction and 15N2 gas incorporation assays. Therefore, we conclude that Herbaspirillum sp. strain B501 is a diazotrophic endophyte compatible with wild rice, particularly O. officinalis. PMID:11679357

  5. The diversity and antimicrobial activity of Preussia sp. endophytes isolated from Australian dry rainforests.

    PubMed

    Mapperson, Rachel R; Kotiw, Michael; Davis, Rohan A; Dearnaley, John D W

    2014-01-01

    Limited knowledge currently exists regarding species diversity and antimicrobial activity of endophytic isolates of Preussia within Australia. This report describes endophytic Preussia species that were identified through molecular analysis of the internal transcribed spacer region. Screening for antimicrobial secondary metabolites was determined by testing crude ethyl acetate (EtOAc) extracts derived from fungal mycelia against a panel of ATCC type strains which included Bacillus cereus, Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, Serratia marcescens, methicillin-resistant Staphylococcus aureus (MRSA) and the opportunist yeast pathogen Candida albicans. Subsequently, high-performance liquid chromatography generated fractions of bioactive EtOAc extracts which were subject to confirmatory testing using the Clinical and Laboratory Standards Institute reference microdilution antimicrobial activity assay. A total of 18 Preussia were isolated from nine host plants with 6/18 having a <97 % sequence similarity to other known species in Genbank, suggesting that they are new species. In preliminary screening, 13/18 Preussia isolates revealed antimicrobial activity against at least one of the microbes tested, whilst 6/18 isolates, including 4/6 putative new species showed specific antimicrobial activity against MRSA and C. albicans. These results highlight the antimicrobial potential of Australian Preussia spp. and also the importance of Australian dry rainforests as an untapped repository of potentially significant bioactive compounds.

  6. Isolation, Purification and Characterization of Vinblastine and Vincristine from Endophytic Fungus Fusarium oxysporum Isolated from Catharanthus roseus

    PubMed Central

    Kumar, Ashutosh; Patil, Deepak; Rajamohanan, Pattuparambil Ramanpillai; Ahmad, Absar

    2013-01-01

    Endophytic fungi reside in a symbiotic fashion inside their host plants, mimic their chemistry and interestingly, produce the same natural products as their hosts and are thus being screened for the production of valuable compounds like taxol, camptothecin, podophyllotoxin, etc. Vinblastine and vincristine are excellent anti-cancer drugs but their current production using plants is non-abundant and expensive. In order to make these drugs readily available to the patients at affordable prices, we isolated the endophytic fungi from Catharanthus roseus plant and found a fungus AA-CRL-6 which produces vinblastine and vincristine in appreciable amounts. These drugs were purified by TLC and HPLC and characterized using UV-Vis spectroscopy, ESI-MS, MS/MS and 1H NMR. One liter of culture filtrate yielded 76 µg and 67 µg of vinblastine and vincristine respectively. This endophytic fungal strain was identified as Fusarium oxysporum based upon its cultural and morphological characteristics and internal transcribed spacer (ITS) sequence analysis. PMID:24066024

  7. Plant Growth Promoting of Endophytic Sporosarcina aquimarina SjAM16103 Isolated from the Pneumatophores of Avicennia marina L.

    PubMed Central

    Janarthine, S. Rylo Sona; Eganathan, P.

    2012-01-01

    Endophytic Sporosarcina aquimarina SjAM16103 was isolated from the inner tissues of pneumatophores of mangrove plant Avicennia marina along with Bacillus sp. and Enterobacter sp. Endophytic S. aquimarina SjAM16103 was Gram variable, and motile bacterium measured 0.6–0.9 μm wide by 1.7–2.0 μm long and light orange-brown coloured in 3-day cultures on tryptone broth at 26°C. Nucleotide sequence of this strain has been deposited in the GenBank under accession number GU930359. This endophytic bacterium produced 2.37 μMol/mL of indole acetic acid and siderophore as it metabolites. This strain could solubilize phosphate molecules and fixes atmospheric nitrogen. Endophytic S. aquimarina SjAM16103 was inoculated into four different plants under in vitro method to analyse its growth-promoting activity and role inside the host plants. The growth of endophytic S. aquimarina SjAM16103 inoculated explants were highly significant than the uninoculated control explants. Root hairs and early root development were observed in the endophytic S. aquimarina SjAM16103 inoculated explants. PMID:22811715

  8. Bioactive metabolites produced by Chaetomium globosum, an endophytic fungus isolated from Ginkgo biloba.

    PubMed

    Qin, Jian-Chun; Zhang, Ya-Mei; Gao, Jin-Ming; Bai, Ming-Sheng; Yang, Sheng-Xiang; Laatsch, Hartmut; Zhang, An-Ling

    2009-03-15

    A novel cytotoxic chlorinated azaphilone derivative named chaetomugilin D (1), together with three known metabolites, chaetomugilin A (2), chaetoglobosins A (3) and C (4), has been isolated by a bioassay-guided fractionation from the EtOAc extract of the cultures of Chaetomium globosum, an endophytic fungus found in the leaves of Ginkgo biloba. Structure of 1 was established by analyses of spectroscopic methods, including 2D-NMR experiments (COSY, NOESY, HMQC, and HMBC). Compounds 1-4 displayed significant growth inhibitory activity against the brine shrimp (Artemia salina) and Mucor miehei.

  9. Phylogenetic reconstruction of endophytic fungal isolates using internal transcribed spacer 2 (ITS2) region.

    PubMed

    GokulRaj, Kathamuthu; Sundaresan, Natesan; Ganeshan, Enthai Jagan; Rajapriya, Pandi; Muthumary, Johnpaul; Sridhar, Jayavel; Pandi, Mohan

    2014-01-01

    Endophytic fungi are inhabitants of plants, living most part of their lifecycle asymptomatically which mainly confer protection and ecological advantages to the host plant. In this present study, 48 endophytic fungi were isolated from the leaves of three medicinal plants and characterized based on ITS2 sequence - secondary structure analysis. ITS2 secondary structures were elucidated with minimum free energy method (MFOLD version 3.1) and consensus structure of each genus was generated by 4SALE. ProfDistS was used to generate ITS2 sequence structure based phylogenetic tree respectively. Our elucidated isolates were belonging to Ascomycetes family, representing 5 orders and 6 genera. Colletotrichum/Glomerella spp., Diaporthae/Phomopsis spp., and Alternaria spp., were predominantly observed while Cochliobolus sp., Cladosporium sp., and Emericella sp., were represented by singletons. The constructed phylogenetic tree has well resolved monophyletic groups with >50% bootstrap value support. Secondary structures based fungal systematics improves not only the stability; it also increases the precision of phylogenetic inference. Above ITS2 based phylogenetic analysis was performed for our 48 isolates along with sequences of known ex-types taken from GenBank which confirms the efficiency of the proposed method. Further, we propose it as superlative marker for reconstructing phylogenetic relationships at different taxonomic levels due to their lesser length.

  10. Isolation and identification of endophytic and mycorrhizal fungi from seeds and roots of Dendrobium (Orchidaceae).

    PubMed

    Chen, Juan; Wang, Hui; Guo, Shun-Xing

    2012-05-01

    The seed germination of orchids under natural conditions requires association with mycorrhizal fungi. Dendrobium nobile and Dendrobium chrysanthum are threatened orchid species in China where they are considered medicinal plants. For conservation and application of Dendrobium using symbiosis technology, we isolated culturable endophytic and mycorrhizal fungi colonized in the protocorms and adult roots of two species plants and identified them by morphological and molecular analyses (5.8S and nrLSU). Of the 127 endophytic fungi isolated, 11 Rhizoctonia-like strains were identified as Tulasnellales (three strains from protocorms of D. nobile), Sebacinales (three strains from roots of D. nobile and two strains from protocorms of D. chrysanthum) and Cantharellales (three strains from roots of D. nobile), respectively. In addition, species of Xylaria, Fusarium, Trichoderma, Colletotrichum, Pestalotiopsis, and Phomopsis were the predominant non-mycorrhizal fungi isolated, and their probable ecological roles in the Dendrobium plants are discussed. These fungal resources will be of great importance for the large-scale cultivation of Dendrobium plants using symbiotic germination technology and for the screening of bioactive metabolites from them in the future. © Springer-Verlag 2012

  11. Nematicidal activity of fervenulin isolated from a nematicidal actinomycete, Streptomyces sp. CMU-MH021, on Meloidogyne incognita.

    PubMed

    Ruanpanun, Pornthip; Laatsch, Hartmut; Tangchitsomkid, Nuchanart; Lumyong, Saisamorn

    2011-06-01

    An isolate of the actinomycete, Streptomyces sp. CMU-MH021 produced secondary metabolites that inhibited egg hatch and increased juvenile mortality of the root-knot nematode Meloidogyne incognita in vitro. 16S rDNA gene sequencing showed that the isolate sequence was 99% identical to Streptomyces roseoverticillatus. The culture filtrates form different culture media were tested for nematocidal activity. The maximal activity against M. incognita was obtained by using modified basal (MB) medium. The nematicidal assay-directed fractionation of the culture broth delivered fervenulin (1) and isocoumarin (2). Fervenulin, a low molecular weight compound, shows a broad range of biological activities. However, nematicidal activity of fervenulin was not previously reported. The nematicidal activity of fervenulin (1) was assessed using the broth microdilution technique. The lowest minimum inhibitory concentrations (MICs) of the compound against egg hatch of M. incognita was 30 μg/ml and juvenile mortality of M. incognita increasing was observed at 120 μg/ml. Moreover, at the concentration of 250 μg/ml fervenulin (1) showed killing effect on second-stage nematode juveniles of M. incognita up to 100% after incubation for 96 h. Isocoumarin (2), another bioactive compound produced by Streptomyces sp. CMU-MH021, showed weak nematicidal activity with M. incognita.

  12. Verrucosispora maris sp. nov., a novel deep-sea actinomycete isolated from a marine sediment which produces abyssomicins.

    PubMed

    Goodfellow, Michael; Stach, James E M; Brown, Roselyn; Bonda, Avinash Naga Venkata; Jones, Amanda L; Mexson, Joanne; Fiedler, Hans-Peter; Zucchi, Tiago Domingues; Bull, Alan T

    2012-01-01

    Verrucosispora isolate AB-18-032(T), the abyssomicin- and proximicin-producing actinomycete, has chemotaxonomic and morphological properties consistent with its classification in the genus Verrucosispora. The organism formed a distinct phyletic line in the Verrucosispora 16S rRNA gene tree sharing similarities of 99.7%, 98.7% and 98.9% with Verrucosispora gifhornensis DSM 44337(T), Verrucosispora lutea YIM 013(T) and Verrucosispora sediminis MS 426(T), respectively. It was readily distinguished from the two latter species using a range of phenotypic features and from V. gifhornensis DSM 44337(T), its nearest phylogenetic neighbor, by a DNA G+C content of 65.5 mol% obtained by thermal denaturation and fluorometry and DNA:DNA relatedness values of 64.0% and 65.0% using renaturation and fluorometric methods, respectively. It is apparent from the combined genotypic and phenotypic data that strain AB-18-032(T) should be classified in the genus Verrucosispora as a new species. The name Verrucosispora maris sp. nov. is proposed for this taxon with isolate AB-18-032(T) (= DSM 45365(T) = NRRL B-24793(T)) as the type strain.

  13. Effectiveness and toxicity of a novel isolated actinomycete strain Streptomyces sp. JS01 on a harmful alga Phaeocystis globosa.

    PubMed

    Zhang, Huajun; Zhang, Su; Peng, Yun; Li, Yi; Cai, Guanjing; Chen, Zhangran; Zheng, Wei; Tian, Yun; Xu, Hong; Zheng, Tianling

    2015-06-01

    An aquatic actinomycete capable of eliminating the brown tide causing marine alga Phaeocystis globosa was isolated from the surface sea water and the isolate named JS01 was characterized as Streptomyces on the basis of its 16S rRNA gene sequence. The supernatant of JS01 could lyse algal cells, implying that JS01 produced a latent alga-lytic compound. Considering this algicidal activity and the response of the algal cells, Chlorophyll a fluorescence decreased significantly in P. globosa in response to the JS01 supernatant when analyzed with flow cytometry. The algal cells experienced cell shrinkage and plasmolysis before disintegration after 72 h of treatment. The released algicide(s) were heat-tolerant, except above 121 °C, and fluctuation in pH variations; even so, algicidal activity was also over 60 %. The maximum toxicity of JS01 was on the seventh day of culture, and the relative luminosity was 0.49 at that time when detected by luminous bacteria Vibrio fischeri. These results indicated that the Streptomyces sp. JS01 could function as a potential controller of Phaeocystis globosa blooms.

  14. Rapid identification of clinically significant species and taxa of aerobic actinomycetes, including Actinomadura, Gordona, Nocardia, Rhodococcus, Streptomyces, and Tsukamurella isolates, by DNA amplification and restriction endonuclease analysis.

    PubMed Central

    Steingrube, V A; Wilson, R W; Brown, B A; Jost, K C; Blacklock, Z; Gibson, J L; Wallace, R J

    1997-01-01

    A previously described PCR-restriction fragment length polymorphism (RFLP) identification schema for Nocardia that used an amplified 439-bp segment (amplicon) of the 65-kDa heat shock protein gene was evaluated for potential use with isolates of all clinically significant aerobic actinomycetes. The study included 28 reference (American Type Culture Collection) strains and 198 clinical isolates belonging to 20 taxonomic groups. Of these 198 isolates, 188 could be differentiated by this PCR-RFLP method. Amplicons from all aerobic actinomycete isolates lacked BstEII recognition sites, thereby distinguishing them from those of mycobacteria that contain one or more such sites. Of 29 restriction endonucleases, MspI plus HinfI produced RFLP patterns that differentiated 16 of the 20 taxa. A single RFLP pattern was observed for 15 of 20 taxa that included 65% of phenotypically clustered isolates. Multiple patterns were seen with Gordona bronchialis, Nocardia asteroides complex type VI, Nocardia otitidiscaviarum, Nocardia transvalensis, and Streptomyces spp. Streptomyces RFLP patterns were the most heterogeneous (five patterns among 19 isolates), but exhibited a unique HinfI fragment of > 320 bp. RFLP patterns that matched those from type strains of Streptomyces albus, Streptomyces griseus, or Streptomyces somaliensis were obtained from 14 of 19 Streptomyces isolates. Only 10 of 28 isolates of N. otitidiscaviarum failed to yield satisfactory amplicons, while only 6 of 188 (3.2%) clinical isolates exhibited patterns that failed to match one of the 21 defined RFLP patterns. These studies extended the feasibility of using PCR-RFLP analysis as a rapid method for the identification of all clinically significant species and taxa of aerobic actinomycetes. PMID:9157134

  15. Rapid identification of clinically significant species and taxa of aerobic actinomycetes, including Actinomadura, Gordona, Nocardia, Rhodococcus, Streptomyces, and Tsukamurella isolates, by DNA amplification and restriction endonuclease analysis.

    PubMed

    Steingrube, V A; Wilson, R W; Brown, B A; Jost, K C; Blacklock, Z; Gibson, J L; Wallace, R J

    1997-04-01

    A previously described PCR-restriction fragment length polymorphism (RFLP) identification schema for Nocardia that used an amplified 439-bp segment (amplicon) of the 65-kDa heat shock protein gene was evaluated for potential use with isolates of all clinically significant aerobic actinomycetes. The study included 28 reference (American Type Culture Collection) strains and 198 clinical isolates belonging to 20 taxonomic groups. Of these 198 isolates, 188 could be differentiated by this PCR-RFLP method. Amplicons from all aerobic actinomycete isolates lacked BstEII recognition sites, thereby distinguishing them from those of mycobacteria that contain one or more such sites. Of 29 restriction endonucleases, MspI plus HinfI produced RFLP patterns that differentiated 16 of the 20 taxa. A single RFLP pattern was observed for 15 of 20 taxa that included 65% of phenotypically clustered isolates. Multiple patterns were seen with Gordona bronchialis, Nocardia asteroides complex type VI, Nocardia otitidiscaviarum, Nocardia transvalensis, and Streptomyces spp. Streptomyces RFLP patterns were the most heterogeneous (five patterns among 19 isolates), but exhibited a unique HinfI fragment of > 320 bp. RFLP patterns that matched those from type strains of Streptomyces albus, Streptomyces griseus, or Streptomyces somaliensis were obtained from 14 of 19 Streptomyces isolates. Only 10 of 28 isolates of N. otitidiscaviarum failed to yield satisfactory amplicons, while only 6 of 188 (3.2%) clinical isolates exhibited patterns that failed to match one of the 21 defined RFLP patterns. These studies extended the feasibility of using PCR-RFLP analysis as a rapid method for the identification of all clinically significant species and taxa of aerobic actinomycetes.

  16. Cryptoendolithic actinomycetes from antarctic sandstone rock samples: Micromonospora endolithica sp. nov. and two isolates related to Micromonospora coerulea Jensen 1932.

    PubMed

    Hirsch, Peter; Mevs, Udo; Kroppenstedt, Reiner M; Schumann, Peter; Stackebrandt, Erko

    2004-03-01

    Three cryptoendolithic, aerobic actinomycetes (AA-459T, AA-319 and AA-321) from antarctic sandstone were characterised phenotypically and by molecular taxonomic methods. The isolates had single spores on substrate mycelium, meso-diaminopimelic acid (m-DAP) and glycine (cell wall type II), a whole cell sugar pattern D (galactose, xylose, arabinose, glucose or rhamnose) and phospholipids of type PII (diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol). Their predominant fatty acids were iso-16:0 and iso-15:0 or 17:1omega8c, the menaquinone profile was complex with mainly MK10 (H4) and MK10 (H6). A wide variety of sugars and several acids were utilised for growth. The isolates were sensitive to a few antibiotics, but formation and excretion of antibiotics was not observed. Phenotypically, isolates AA-319 and AA-321 were similar. Phylogenetic analysis of 16S rRNA gene sequences revealed close relationship of strains AA-319 and AA-321 with each other (99.5%) and clustering (98.5%) with Micromonospora coerulea DSM 43143T. DNA-DNA hybridisation showed both strains to be genomically highly similar to strain DSM 43143T. Phenotypically they could be viewed as separate taxa, but presently they will be considered as strains of Micromonospora coerulea. Strain AA-459T was phylogenetically close to Micromonospora chersina DSM 44151T (99.1%) and to Micromonospora rosaria DSM 803T, but DNA-DNA similarity with M. chersina DSM 44151T was low with 28.9/33.5 %, indicating the presence of a different and new species. Consequently, isolate AA-459T (DSM 44398T NRRL B-24248T) is described as the type strain of Micromonospora endolithica sp. nov.

  17. Decolourisation of Synthetic Dyes by Endophytic Fungal Flora Isolated from Senduduk Plant (Melastoma malabathricum)

    PubMed Central

    Ngieng, Ngui Sing

    2013-01-01

    A total of twenty endophytic fungi successfully isolated from Melastoma malabathricum (Senduduk) were examined for their ability to decolourise azo dyes: Congo red, Orange G, and Methyl red and an anthraquinone dye, Remazol Brilliant Blue R. Initial screening on the glucose minimal media agar plates amended with 200 mg L−1 of each respective dye showed that only isolate MS8 was able to decolourise all of the four dyes. The isolate decolourised completely both the RBBR and Orange G in the agar medium within 8 days. Further quantitative analysis of the dye decolourisation by isolate MS8 in aqueous minimal medium showed that isolate MS8 was able to decolourise all the tested dyes at varying levels. Dye decolourisation by the isolate MS8 was determined to be 97% for RBBR, 33% for Orange G, 48% for Congo red, and 56% for Methyl red, respectively, within a period of 16 days. Molecular identification of the fungal isolate MS8 using primer ITS1 and ITS4 showed that isolate MS8 shared 99% sequence similarity with Marasmius cladophyllus, a Basidiomycete. The ability to decolourise different types of dyes by isolate MS8 thus suggested a possible application of this fungus in the decolourisation of dyestuff effluents. PMID:25937973

  18. Gancidin W, a potential low-toxicity antimalarial agent isolated from an endophytic Streptomyces SUK10

    PubMed Central

    Zin, Noraziah Mohamad; Baba, Mohd Shukri; Zainal-Abidin, Abu Hassan; Latip, Jalifah; Mazlan, Noor Wini; Edrada-Ebel, RuAngelie

    2017-01-01

    Endophytic Streptomyces strains are potential sources for novel bioactive molecules. In this study, the diketopiperazine gancidin W (GW) was isolated from the endophytic actinobacterial genus Streptomyces, SUK10, obtained from the bark of Shorea ovalis tree, and it was tested in vivo against Plasmodium berghei PZZ1/100. GW exhibited an inhibition rate of nearly 80% at 6.25 and 3.125 μg kg−1 body weight on day four using the 4-day suppression test method on male ICR strain mice. Comparing GW at both concentrations with quinine hydrochloride and normal saline as positive and negative controls, respectively, 50% of the mice treated with 3.125 μg kg−1 body weight managed to survive for more than 11 months after infection, which almost reached the life span of normal mice. Biochemical tests of selected enzymes and proteins in blood samples of mice treated with GW were also within normal levels; in addition, no abnormalities or injuries were found on internal vital organs. These findings indicated that this isolated bioactive compound from Streptomyces SUK10 exhibits very low toxicity and is a good candidate for potential use as an antimalarial agent in an animal model. PMID:28223778

  19. Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity.

    PubMed

    Pan, Feng; Liu, Zheng-Qiong; Chen, Que; Xu, Ying-Wen; Hou, Kai; Wu, Wei

    2016-01-01

    The aim of this paper is to identify and investigate an endophytic fungus (strain 28) that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc) extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass.) Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB) as the base culture medium, fermentation for 4-8d (initial pH: 7.5), followed by extraction with EtOAc. The extract was stable at temperatures up to 80°C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  20. Biological activities from extracts of endophytic fungi isolated from Viguiera arenaria and Tithonia diversifolia.

    PubMed

    Guimarães, Denise O; Borges, Warley S; Kawano, Cristina Y; Ribeiro, Patrícia H; Goldman, Gustavo H; Nomizo, Auro; Thiemann, Otávio H; Oliva, Glaucius; Lopes, Norberto P; Pupo, Mônica T

    2008-01-01

    A total of 39 endophytic fungi have been isolated from Viguiera arenaria and Tithonia diversifolia, both collected in São Paulo State, Brazil. The isolates were identified based on their ribosomal DNA sequences. The ethyl acetate (EtOAc) extracts of all endophytic fungi were evaluated for their antimicrobial, antiparasitic and antitumoral activity. Antimicrobial screening was conducted using an agar diffusion assay against three pathogenic microorganisms: Staphylococcus aureus, Escherichia coli and Candida albicans. Antiparasitic activity was determined by enzymatic inhibition of gGAPDH of Trypanosoma cruzi and adenine phosphorybosiltransferase (APRT) of Leishmania tarentolae. Antitumoral activity was tested against human T leukemia cells by the Mosmann colorimetric method. All extracts showed activity in at least one assay: 79.5% of the extracts were cytotoxic against leukemia cells, 5.1% of the extracts were active against S. aureus, 25.6% against E. coli and 64.1% against Candida albicans. Only one extract showed promising results in the inhibition of parasitic enzymes gGAPDH (95.0%) and three were found to inhibit APRT activity. The cytotoxic extract produced by the strain VA1 (Glomerella cingulata) was fractionated and yielded nectriapyrone and tyrosol. Nectriapyrone showed relevant cytotoxic activity against both human T leukemia and melanoma tumor cell lines.

  1. Gancidin W, a potential low-toxicity antimalarial agent isolated from an endophytic Streptomyces SUK10.

    PubMed

    Zin, Noraziah Mohamad; Baba, Mohd Shukri; Zainal-Abidin, Abu Hassan; Latip, Jalifah; Mazlan, Noor Wini; Edrada-Ebel, RuAngelie

    2017-01-01

    Endophytic Streptomyces strains are potential sources for novel bioactive molecules. In this study, the diketopiperazine gancidin W (GW) was isolated from the endophytic actinobacterial genus Streptomyces, SUK10, obtained from the bark of Shorea ovalis tree, and it was tested in vivo against Plasmodium berghei PZZ1/100. GW exhibited an inhibition rate of nearly 80% at 6.25 and 3.125 μg kg(-1) body weight on day four using the 4-day suppression test method on male ICR strain mice. Comparing GW at both concentrations with quinine hydrochloride and normal saline as positive and negative controls, respectively, 50% of the mice treated with 3.125 μg kg(-1) body weight managed to survive for more than 11 months after infection, which almost reached the life span of normal mice. Biochemical tests of selected enzymes and proteins in blood samples of mice treated with GW were also within normal levels; in addition, no abnormalities or injuries were found on internal vital organs. These findings indicated that this isolated bioactive compound from Streptomyces SUK10 exhibits very low toxicity and is a good candidate for potential use as an antimalarial agent in an animal model.

  2. Cytotoxic and Antifungal Constituents Isolated from the Metabolites of Endophytic Fungus DO14 from Dendrobium officinale.

    PubMed

    Wu, Ling-Shang; Jia, Min; Chen, Ling; Zhu, Bo; Dong, Hong-Xiu; Si, Jin-Ping; Peng, Wei; Han, Ting

    2015-12-22

    Two novel cytotoxic and antifungal constituents, (4S,6S)-6-[(1S,2R)-1, 2-dihydroxybutyl]-4-hydroxy-4-methoxytetrahydro-2H-pyran-2-one (1), (6S,2E)-6-hydroxy-3-methoxy-5-oxodec-2-enoic acid (2), together with three known compounds, LL-P880γ (3), LL-P880α (4), and Ergosta-5,7,22-trien-3b-ol (5) were isolated from the metabolites of endophytic fungi from Dendrobium officinale. The chemical structures were determined based on spectroscopic methods. All the isolated compounds 1-5 were evaluated by cytotoxicity and antifungal effects. Our present results indicated that compounds 1-4 showed notable anti-fungal activities (minimal inhibitory concentration (MIC) ≤ 50 μg/mL) for all the tested pathogens including Candida albicans, Cryptococcus neoformans, Trichophyton rubrum, Aspergillus fumigatus. In addition, compounds 1-4 possessed notable cytotoxcities against human cancer cell lines of HL-60 cells with the IC50 values of below 100 μM. Besides, compounds 1, 2, 4 and 5 showed strong cytotoxities on the LOVO cell line with the IC50 values were lower than 100 μM. In conclusion, our study suggested that endophytic fungi of D. officinale are great potential resources to discover novel agents for preventing or treating pathogens and tumors.

  3. Novel bioactive compounds from actinomycetes.

    PubMed

    Sanglier, J J; Wellington, E M; Behal, V; Fiedler, H P; Ellouz Ghorbel, R; Finance, C; Hacene, M; Kamoun, A; Kelly, C; Mercer, D K

    1993-10-01

    Actinomycetes form an enormous reservoir of secondary metabolites and enzymes. The potential for exploiting rare actinomycetes is highlighted by the discovery of novel compounds from strains of Spirillospora and Nocardioides. Novel compounds of well known classes of antibiotics, such as polyenes, continue to be discovered. For compounds containing a chromophore, the analysis by high-performance liquid chromatography coupled with a diode-array detector enables the elimination of producers of known compounds and facilitates the discovery of novel compounds or derivatives. The complexity of the regulatory mechanisms is illustrated by glutamine synthetase. The characterization of thermostable amylolytic, lignolytic, peroxidase and neuramidase activities, and the isolation of novel cellulolytic actinomycetes clearly demonstrate the potential of Actinomycetes as producers of enzymes.

  4. Endophytic Bacteria Isolated from Common Bean (Phaseolus vulgaris) Exhibiting High Variability Showed Antimicrobial Activity and Quorum Sensing Inhibition.

    PubMed

    Lopes, Ralf Bruno Moura; Costa, Leonardo Emanuel de Oliveira; Vanetti, Maria Cristina Dantas; de Araújo, Elza Fernandes; de Queiroz, Marisa Vieira

    2015-10-01

    Endophytic bacteria play a key role in the biocontrol of phytopathogenic microorganisms. In this study, genotypic diversity was analyzed via repetitive element PCR (rep-PCR) of endophytic isolates of the phylum Actinobacteria that were previously collected from leaves of cultivars of common bean (Phaseolus vulgaris). Considerable variability was observed, which has not been reported previously for this phylum of endophytic bacteria of the common bean. Furthermore, the ethanol extracts from cultures of various isolates inhibited the growth of pathogenic bacteria in vitro, especially Gram-positive pathogens. Extracts from cultures of Microbacterium testaceum BAC1065 and BAC1093, which were both isolated from the 'Talismã' cultivar, strongly inhibited most of the pathogenic bacteria tested. Bean endophytic bacteria were also demonstrated to have the potential to inhibit the quorum sensing of Gram-negative bacteria. This mechanism may regulate the production of virulence factors in pathogens. The ability to inhibit quorum sensing has also not been reported previously for endophytic microorganisms of P. vulgaris. Furthermore, M. testaceum with capacity to inhibit quorum sensing appears to be widespread in common bean. The genomic profiles of M. testaceum were also analyzed via pulsed-field gel electrophoresis, and greater differentiation was observed using this method than rep-PCR; in general, no groups were formed based on the cultivar of origin. This study showed for the first time that endophytic bacteria from common bean plants exhibit high variability and may be useful for the development of strategies for the biological control of diseases in this important legume plant.

  5. Nonomuraea monospora sp. nov., an actinomycete isolated from cave soil in Thailand, and emended description of the genus Nonomuraea.

    PubMed

    Nakaew, Nareeluk; Sungthong, Rungroch; Yokota, Akira; Lumyong, Saisamorn

    2012-12-01

    A novel actinomycete, designated strain PT708(T), was isolated from cave soil collected in Pha Tup Cave Forest Park, Nan province, Thailand. It produced compounds with antimicrobial and anticancer activities. Its chemotaxonomic properties were consistent with those of members of the genus Nonomuraea. The major menaquinone was MK-9(H(4)), with minor amounts of MK-9(H(6)), MK-9(H(2)), MK-10(H(2)) and MK-8(H(4)). The polar lipid profile contained phosphatidylmonomethylethanolamine, diphosphatidylglycerol, hydroxy-phosphatidylmonomethylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol mannoside and phosphatidylinositol. The major fatty acids were iso-C(16:0), 10-methyl C(17:0), C(16:0) and C(17:1)ω6c. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain PT708(T) belonged to the genus Nonomuraea and was most closely related to Nonomuraea rhizophila YIM 67092(T) (98.50% sequence similarity) and Nonomuraea rosea GW 12687(T) (98.30%). The genomic DNA G+C content of strain PT708(T) was 73.3 mol%. Unlike the recognized members of the genus Nonomuraea, the novel strain formed single spores at the tips of aerial hyphae. Based on the phenotypic, phylogenetic and genotypic evidence, strain PT708(T) represents a novel species of the genus Nonomuraea, for which the name Nonomuraea monospora sp. nov. is proposed. The type strain is PT708(T) ( = TISTR 1910(T) = JCM 16114(T)).

  6. Streptomyces lasiicapitis sp. nov., an actinomycete that produces kanchanamycin, isolated from the head of an ant (Lasius fuliginosus L.).

    PubMed

    Ye, Lan; Zhao, Shanshan; Li, Yao; Jiang, Shanwen; Zhao, Yue; Li, Jinmeng; Yan, Kai; Wang, Xiangjing; Xiang, Wensheng; Liu, Chongxi

    2017-05-01

    During a screening for novel and biotechnologically useful actinobacteria in insects, a kanchanamycin-producing actinomycete with antifungal activity, designated strain 3H-HV17(2)T, was isolated from the head of an ant (Lasius fuliginosus L.) and characterized using a polyphasic approach. 16S rRNA gene sequence similarity studies showed that strain 3H-HV17(2)T belongs to the genus Streptomyces with the highest sequence similarities to Streptomyces spectabilis NBRC 13424T (98.90 %, with which it phylogenetically clustered, Streptomyces alboflavus NRRL B-2373T (98.65 %) and Streptomyces flavofungini NBRC 13371T (98.36 %). Phylogenetic analysis based on the gyrB gene also supported the close relationship of these strains. The morphological and chemotaxonomic properties of the strain are also consistent with those members of the genus Streptomyces. A combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 3H-HV17(2)T and its phylogenetically closely related strains, which further clarified their relatedness and demonstrated that strain 3H-HV17(2)T could be distinguished from these strains. Therefore, strain 3H-HV17(2)T is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces lasiicapitis sp. nov. is proposed. The type strain is 3H-HV17(2)T (=CGMCC 4.7349T=DSM 103124T).

  7. Complete Genome Sequence of Kosakonia sacchari Strain BO-1, an Endophytic Diazotroph Isolated from a Sweet Potato

    PubMed Central

    Shinjo, Rina; Uesaka, Kazuma; Ihara, Kunio; Loshakova, Kseniia; Mizuno, Yuri; Yano, Katsuya

    2016-01-01

    The complete genome sequence of the endophytic diazotroph Kosakonia sacchari, isolated from a sweet potato, was analyzed. The 4,902,106-bp genome with 53.7% G+C content comprises 4,638 open reading frames, including nif genes, 84 tRNAs, and seven complete rRNAs in a circular chromosome. PMID:27609910

  8. Complete Genome Sequences of the Endophytic Streptomyces Strains EN16, EN23, and EN27, Isolated from Wheat Plants

    PubMed Central

    Araujo, Ricardo; Adetutu, Eric; Tobe, Shanan S.; Mallya, Sandeep; Paul, Bobby; Satyamoorthy, Kapaettu

    2016-01-01

    The complete genome sequences of three endophytic Streptomyces species were compared. Strains EN16, EN23, and EN27 were isolated from surface-sterilized roots of wheat plants from South Australia. In field trials, these strains are effective in suppressing fungal root diseases of wheat when added as spore coatings to wheat seed. PMID:27932645

  9. Draft Genome Sequence of Enterobacter sp. Sa187, an Endophytic Bacterium Isolated from the Desert Plant Indigofera argentea

    PubMed Central

    Lafi, Feras F.; Alam, Intikhab; Geurts, Rene; Bisseling, Ton; Bajic, Vladimir B.

    2017-01-01

    ABSTRACT Enterobacter sp. Sa187 is a plant endophytic bacterium, isolated from root nodules of the desert plant Indigofera argentea, collected from the Jizan region of Saudi Arabia. Here, we report the genome sequence of Sa187, highlighting several genes involved in plant growth–promoting activity and environmental adaption. PMID:28209831

  10. Isolation of endophytic endospore-forming bacteria from Theobroma cacao as potential biological control agents of cacao dieseases

    USDA-ARS?s Scientific Manuscript database

    Endospore-forming bacterial endophytes were isolated from Theobroma cacao to access the present and diversity of endospore-forming bacteria in cacao. Cacao leaves, pods, branches, and flower cushions were removed from cacao trees escaping disease on INIAP’s Tropical Research Station in Pichilingue, ...

  11. Expression of Russian Wheat Aphid (Homoptera: Aphididae) Resistance in Genotypes of Tall Fescue Harboring Different Isolates of Acremonium Endophyte

    Treesearch

    S.L. Clement; D.G. Lester; A. Dan Wilson; R.C. Johnson; J.H. Bouton

    1996-01-01

    Experiments were conducted to compare the expression of Russian wheat aphid, Diurnphis noxia (Mordvilko), resistance in 2 genotypes of tall fescue grass, Festucn arundinacea Schreb., harboring different isolates of the endophytic fungus Acremonium coenophialum Morgan-Jones & cams. Aphids did not select...

  12. Draft Genome Sequence of Pseudomonas putida Strain GM4FR, an Endophytic Bacterium Isolated from Festuca rubra L.

    PubMed Central

    Hollensteiner, Jacqueline; Granzow, Sandra; Daniel, Rolf; Vidal, Stefan; Wemheuer, Bernd

    2017-01-01

    ABSTRACT Pseudomonas putida GM4FR is an endophytic bacterium isolated from aerial plant tissues of Festuca rubra L. Functional annotation of the draft genome (7.1 Mb) revealed 6,272 predicted protein-encoding genes. The genome provides insights into the biocontrol and plant growth-promoting potential of P. putida GM4FR. PMID:28360162

  13. Interaction between a dark septate endophytic isolate from Dendrobium sp. and roots of D. nobile seedlings.

    PubMed

    Hou, Xiao-Qiang; Guo, Shun-Xing

    2009-04-01

    Interactions between an isolate of dark septate endophytes (DSE) and roots of Dendrobium nobile Lindl. seedlings are reported in this paper. The isolate was obtained from orchid mycorrhizas on Dendrobium sp. in subtropical forest. The fungus formed typical orchid mycorrhiza in aseptic co-culture with D. nobile seedlings on modified Murashige-Skoog (MMS) medium. Anatomic observations of the infected roots showed that the DSE hyphae invaded the velamen layer, passed through passage cells in exodermis, entered the cortex cells, and then formed fungal pelotons of orchid mycorrhiza. D. nobile seedlings' plant height, stem diameter, new roots number and biomass were greatly enhanced by inoculating the fungus to seedlings. The fungus was identified as Leptodontidium by sequencing the polymerase chain reaction-amplified rDNA ITS1-5.8S-ITS2 (internal transcribed spacer (ITS)) regions and comparison with similar taxa.

  14. Papaya shoot tip associated endophytic bacteria isolated from in vitro cultures and host-endophyte interaction in vitro and in vivo.

    PubMed

    Thomas, Pious; Kumari, Sima; Swarna, Ganiga K; Gowda, T K S

    2007-03-01

    Fourteen distinct bacterial clones were isolated from surface-sterilized shoot tips (approximately 1 cm) of papaya (Carica papaya L. 'Surya') planted on Murashige and Skoog (MS)-based papaya culture medium (23/50 nos.) during the 2-4 week period following in vitro culturing. These isolates were ascribed to six Gram-negative genera, namely Pantoea (P. ananatis), Enterobacter (E. cloacae), Brevundimonas (B. aurantiaca), Sphingomonas, Methylobacterium (M. rhodesianum), and Agrobacterium (A. tumefaciens) or two Gram-positive genera, Microbacterium (M. esteraromaticum) and Bacillus (B. benzoevorans) based on 16S rDNA sequence analysis. Pantoea ananatis was the most frequently isolated organism (70% of the cultures) followed by B. benzoevorans (13%), while others were isolated from single stocks. Bacteria-harboring in vitro cultures often showed a single organism. Pantoea, Enterobacter, and Agrobacterium spp. grew actively on MS-based normal papaya medium, while Microbacterium, Brevundimonas, Bacillus, Sphingomonas, and Methylobacterium spp. failed to grow in the absence of host tissue. Supplying MS medium with tissue extract enhanced the growth of all the organisms in a dose-dependent manner, indicating reliance of the endophyte on its host. Inoculation of papaya seeds with the endophytes (20 h at OD550=0.5) led to delayed germination or slow seedling growth initially. However, the inhibition was overcome by 3 months and the seedlings inoculated with Pantoea, Microbacterium, or Sphingomonas spp. displayed significantly better root and shoot growths.

  15. Sphingomonaszeicaulis sp. nov., an endophytic bacterium isolated from maize root.

    PubMed

    Gao, Jun-Lian; Sun, Pengbo; Wang, Xu-Ming; Cheng, Shoutao; Lv, Fanyang; Qiu, Tian-Lei; Yuan, Mei; Sun, Jian-Guang

    2016-09-01

    A novel Gram-staining-negative, aerobic and rod-shaped strain designated 541T was isolated from surface-sterilized root tissue of maize, collected from the Fangshan District of Beijing, People's Republic of China, and was subjected to a taxonomic study using a polyphasic approach. According to a phylogenetic tree based on 16S rRNA gene sequences, strain 541T represented a member of the genus Sphingomonas and clustered with Sphingomonas sanxanigenens DSM 19645T, with which it shared the highest 16S rRNA gene sequence similarity (98.8 %). The predominant respiratory quinone was ubiquinone-10 (Q-10), the major polyamine was sym-homospermidine and the major cellular fatty acids were C18 : 1ω7c (50.9 %), C16 : 0 (22.0 %) and C14 : 0 2-OH (11.4 %). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. The DNA G+C content was 64.7 mol%. DNA-DNA relatedness between strain 541T and its closest phylogenetic relative Sphingomonas sanxanigenens DSM 19645T was 50.8 %. The results of physiological and biochemical tests and the differences in the fatty acid profiles allowed a clear phenotypic differentiation of strain 541T from closely related species of the genus Sphingomonas. Strain 541T represents a novel species within the genus Sphingomonas, for which the nameSphingomonas zeicaulis sp. nov. is proposed, with the type strain 541T (=CGMCC 1.15008T=DSM 100587T).

  16. Cytotoxic effects of oosporein isolated from endophytic fungus Cochliobolus kusanoi

    PubMed Central

    Ramesha, Alurappa; Venkataramana, M.; Nirmaladevi, Dhamodaran; Gupta, Vijai K.; Chandranayaka, S.; Srinivas, Chowdappa

    2015-01-01

    In the present study, oosporein, a fungal toxic secondary metabolite known to be a toxic agent causing chronic disorders in animals, was isolated from fungus Cochliobolus kusanoi of Nerium oleander L. Toxic effects of oosporein and the possible mechanisms of cytotoxicity as well as the role of oxidative stress in cytotoxicity to Madin-Darby canine kidney kidney cells and RAW 264.7 splene cells were evaluated in vitro. Also to know the possible in vivo toxic effects of oosporein on kidney and spleen, Balb/C mouse were treated with different concentrations of oosporein ranging from 20 to 200 μM). After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs. Oosporein induced elevated levels of reactive oxygen species (ROS) generation and high levels of malondialdehyde, loss of mitochondrial membrane potential, induced glutathione hydroxylase (GSH) production was observed in a dose depended manner. Effects oosporein on chromosomal DNA damage was assessed by Comet assay, and increase in DNA damage were observed in both the studied cell lines by increasing the oosporein concentration. Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutase1 and Catalase ) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay. Histopathological examination of oosporein treated mouse kidney and splenocytes further revealed that, oosporein treated target mouse tissues were significantly damaged with that of untreated sam control mice and these effects were in directly proportional to the the toxin dose. Results of the present study reveals that, ROS is the principle event prompting increased oosporein toxicity in studied in vivio and in vitro animal models. The high previlance of these fungi in temperate climates further

  17. Endophytic and pathogenic isolates of the cacao fungal pathogen Moniliophthora perniciosa (Tricholomataceae) are indistinguishable based on genetic and physiological analysis.

    PubMed

    Lana, T G; Azevedo, J L; Pomella, A W V; Monteiro, R T R; Silva, C B; Araújo, W L

    2011-02-22

    We evaluated the genetic and physiological variability of Moniliophthora perniciosa obtained from healthy and diseased branches of cacao (Theobroma cacao) plants. The diversity of the isolates was evaluated by RAPD technique and by studies of virulence and exoenzyme production. The genetic variability of endophytic and pathogenic M. perniciosa was evaluated in association with pathogenicity assays. RAPD analysis showed eight genetic groups, which were not related to plant disease status (healthy versus diseased branches). Isolates from cacao were included in three groups, excluding isolates from other host plants. Pathogenicity and enzyme analysis showed that the virulence of the isolates is not related to exoenzyme production. This is the first evidence that M. perniciosa colonizes healthy parenchymatic tissues, showing that endophytic behavior may occur in this species.

  18. Anti-phytopathogen potential of endophytic actinobacteria isolated from tomato plants (Lycopersicon esculentum) in southern Brazil, and characterization of Streptomyces sp. R18(6), a potential biocontrol agent.

    PubMed

    de Oliveira, Margaroni Fialho; da Silva, Mariana Germano; Van Der Sand, Sueli T

    2010-09-01

    Tomato plants (Lycopersicon esculentum) are highly susceptible to phytopathogen attack. The resulting intensive application of pesticides on tomato crops can affect the environment and health of humans and animals. The objective of this study was to select potential biocontrol agents among actinobacteria from tomato plants, in a search for alternative phytopathogen control. We evaluated 70 endophytic actinobacteria isolated from tomato plants in southern Brazil, testing their antimicrobial activity, siderophore production, indoleacetic acid production, and phosphate solubility. The actinomycete isolate with the highest antimicrobial potential was selected using the agar-well diffusion method, in order to optimize conditions for the production of compounds with antimicrobial activity. For this study, six growth media (starch casein-SC, ISP2, Bennett's, Sahin, Czapek-Dox, and TSB), three temperatures (25 degrees C, 30 degrees C, and 35 degrees C) and different pH were tested. Of the actinobacteria tested, 88.6% showed antimicrobial activity against at least one phytopathogen, 72.1% showed a positive reaction for indoleacetic acid production, 86.8% produced siderophores and 16.2% showed a positive reaction for phosphate solubility. Isolate R18(6) was selected due to its antagonistic activity against all phytopathogenic microorganisms tested in this study. The best conditions for production were observed in the SC medium, at 30 degrees C and pH 7.0. The isolate R18(6) showed close biochemical and genetic similarity to Streptomyces pluricolorescens. Copyright 2010 Elsevier Masson SAS. All rights reserved.

  19. Isolation of (-)-avenaciolide as the antifungal and antimycobacterial constituent of a Seimatosporium sp. Endophyte from the medicinal plant Hypericum perforatum .

    PubMed

    Clark, Trevor N; Bishop, Amanda I; McLaughlin, Mark; Calhoun, Larry A; Johnson, John A; Gray, Christopher A

    2014-10-01

    An extract of Seimatosporium sp., an endophyte from the Canadian medicinal plant Hypericum perforatum, exhibited significant antifungal and antimycobacterial activity against Candida albicans and Mycobacterium tuberculosis H37Ra. Bioassay guided fractionation led to the isolation of (-)-avenaciolide as the only bioactive constituent of the extract. This is the first report of both the antimycobacterial activity of avenaciolide and its isolation from a Seimatosporium sp. fungus.

  20. Tsukamurella pseudospumae sp. nov., a novel actinomycete isolated from activated sludge foam.

    PubMed

    Nam, Sun-Woo; Kim, Wonyong; Chun, Jongsik; Goodfellow, Michael

    2004-07-01

    The taxonomic position of two Tsukamurella strains isolated from activated sludge foam was clarified. The organisms, isolates JC85 and N1176T, were found to have chemical and morphological properties typical of members of the genus Tsukamurella. DNA-DNA relatedness studies showed that the strains formed a distinct genomic species that was most closely related to Tsukamurella spumae. The two isolates also share a range of phenotypic properties that distinguishes them from representatives of all species of Tsukamurella with validly published names. It is evident from the data that the two organisms should be classified as a novel Tsukamurella species, Tsukamurella pseudospumae sp. nov. The type strain is N1176T (=DSM 44118T=NCIMB 13963T).

  1. Bioactive polyketides and alkaloids from Penicillium citrinum, a fungal endophyte isolated from Ocimum tenuiflorum.

    PubMed

    Lai, Daowan; Brötz-Oesterhelt, Heike; Müller, Werner E G; Wray, Victor; Proksch, Peter

    2013-12-01

    Chemical investigation of the endophytic fungus Penicillium citrinum cultured on white beans or on rice led to the isolation of two new alkaloids (1 and 2), along with fourteen known polyketides (6-12, 14-20) and four known alkaloids (3-5, and 13). The structures of the isolated compounds were determined by extensive analysis of the 1D, 2D NMR, and MS data, and by comparison with the literature. Compound 13, which had been previously obtained only by chemical synthesis, was isolated as a natural product for the first time, while compound 6 was firstly reported as a fungal metabolite. A re-isolation of sclerotinin A (14) revealed it to be a diastereoisomeric mixture (14a and 14b), whose stereochemistry was proposed for the first time based on ROESY experiment. All isolated compounds were evaluated for their cytotoxic and antibacterial activities. Compounds 12 and 17 showed significant cytotoxicity against the murine lymphoma cell line L5178Y with IC50 values of 1.0, and 0.78 μg/ml, respectively, while compounds 5, 11, and 15 were moderately active against Staphylococcus aureus ATCC 29213 (MIC 64 μg/ml).

  2. Congolese Rhizospheric Soils as a Rich Source of New Plant Growth-Promoting Endophytic Piriformospora Isolates

    PubMed Central

    Venneman, Jolien; Audenaert, Kris; Verwaeren, Jan; Baert, Geert; Boeckx, Pascal; Moango, Adrien M.; Dhed’a, Benoît D.; Vereecke, Danny; Haesaert, Geert

    2017-01-01

    In the last decade, there has been an increasing focus on the implementation of plant growth-promoting (PGP) organisms as a sustainable option to compensate for poor soil fertility conditions in developing countries. Trap systems were used in an effort to isolate PGP fungi from rhizospheric soil samples collected in the region around Kisangani in the Democratic Republic of Congo. With sudangrass as a host, a highly conducive environment was created for sebacinalean chlamydospore formation inside the plant roots resulting in a collection of 51 axenically cultured isolates of the elusive genus Piriformospora (recently transferred to the genus Serendipita). Based on morphological data, ISSR fingerprinting profiles and marker gene sequences, we propose that these isolates together with Piriformospora williamsii constitute a species complex designated Piriformospora (= Serendipita) ‘williamsii.’ A selection of isolates strongly promoted plant growth of in vitro inoculated Arabidopsis seedlings, which was evidenced by an increase in shoot fresh weight and a strong stimulation of lateral root formation. This isolate collection provides unprecedented opportunities for fundamental as well as translational research on the Serendipitaceae, a family of fungal endophytes in full expansion. PMID:28261171

  3. A primary assessment of the endophytic bacterial community in a xerophilous moss (Grimmia montana) using molecular method and cultivated isolates.

    PubMed

    Liu, Xiao Lei; Liu, Su Lin; Liu, Min; Kong, Bi He; Liu, Lei; Li, Yan Hong

    2014-01-01

    Investigating the endophytic bacterial community in special moss species is fundamental to understanding the microbial-plant interactions and discovering the bacteria with stresses tolerance. Thus, the community structure of endophytic bacteria in the xerophilous moss Grimmia montana were estimated using a 16S rDNA library and traditional cultivation methods. In total, 212 sequences derived from the 16S rDNA library were used to assess the bacterial diversity. Sequence alignment showed that the endophytes were assigned to 54 genera in 4 phyla (Proteobacteria, Firmicutes, Actinobacteria and Cytophaga/Flexibacter/Bacteroids). Of them, the dominant phyla were Proteobacteria (45.9%) and Firmicutes (27.6%), the most abundant genera included Acinetobacter, Aeromonas, Enterobacter, Leclercia, Microvirga, Pseudomonas, Rhizobium, Planococcus, Paenisporosarcina and Planomicrobium. In addition, a total of 14 species belonging to 8 genera in 3 phyla (Proteobacteria, Firmicutes, Actinobacteria) were isolated, Curtobacterium, Massilia, Pseudomonas and Sphingomonas were the dominant genera. Although some of the genera isolated were inconsistent with those detected by molecular method, both of two methods proved that many different endophytic bacteria coexist in G. montana. According to the potential functional analyses of these bacteria, some species are known to have possible beneficial effects on hosts, but whether this is the case in G. montana needs to be confirmed.

  4. Isolation and characterization of endophytic Bacillus subtilis Jaas ed1 antagonist of eggplant Verticillium wilt.

    PubMed

    Lin, Ling; Qiao, Yong-Sheng; Ju, Zheng-Ying; Ma, Chang-Wen; Liu, Ying-Hao; Zhou, Yi-Jun; Dong, Han-Song

    2009-07-01

    An endophytic bacterial strain, Jaas ed1, was isolated from the interior of eggplant (Solanum melongena L.) stems in Jiangsu Province, China. According to the morphological and physiological characteristics and phylogenetic analysis of the 16s rDNA sequence, it was identified as Bacillius subtilis. Strain Jaas ed1 and its cell-free filtrate had strong antifungal activity against Verticillium dahliae. The strain was an internal colonizer within the eggplant without any harmful side effects to the plant. In greenhouse experiments, the strain cell suspension effectively controlled Verticillium wilt of eggplant, and its control efficiency was far more significant than that of the cell-free filtrate after inoculation of V. dahliae.

  5. [Isolation of endophytic antagonistic bacterium from Amorphophallus konjac and research on its antibacterial metabolite].

    PubMed

    Zhou, Ying; Chen, Lin; Chai, Xin-Li; Yu, Zi-Niu; Sun, Ming

    2007-12-01

    An endophytic antagonistic bacterium was isolated from Amorphophallus konjac calli. In order to identify this bacterium, 16S rDNA was amplified and partially sequenced. Sequence comparison showed that this sequence has the highest similarity to that in Bacillus subtilis, with 99.0% identities. That demonstrated this bacterium belongs to Bacillus subtili , named BSn5. The extracted extracellular protein from strain BSn5 had antibacterial activity against Erwinia carotovora subp. carotovora, which was unstable after heated, sensitive to proteinase K and resistant to trypsin. There was only a 31.6kDa protein component as by SDS-PAGE detection. Nondenaturing polyacrylaminde gel was used to purify this protein. The purified 31.6kDa protein exhibited inhibitory activity against Erwinia carotovora subp. carotovora. This protein is different from all known metabolites from Bacillus subtilis, suggesting that it may be a novel antibacterial protein.

  6. Cytotoxic norsesquiterpene peroxides from the endophytic fungus Talaromyces flavus isolated from the mangrove plant Sonneratia apetala.

    PubMed

    Li, Hanxiang; Huang, Hongbo; Shao, Changlun; Huang, Huarong; Jiang, Jieyi; Zhu, Xun; Liu, Yayue; Liu, Lan; Lu, Yongjun; Li, Mengfeng; Lin, Yongcheng; She, Zhigang

    2011-05-27

    Four new norsesquiterpene peroxides, named talaperoxides A-D (1-4), as well as one known analogue, steperoxide B (5, or merulin A), have been isolated from a mangrove endophytic fungus, Talaromyces flavus. Their structures were elucidated mainly by 1D and 2D NMR. Structures of 1, 2, and 5 were further confirmed by single-crystal X-ray diffraction, and their absolute configurations were also determined using copper radiation. Cytotoxic activities of compounds 1-5 were evaluated in vitro against human cancer cell lines MCF-7, MDA-MB-435, HepG2, HeLa, and PC-3. Compounds 2 and 4 showed cytotoxicity against the five human cancer cell lines with IC50 values between 0.70 and 2.78 μg/mL.

  7. Diverse endophytic bacteria isolated from a leguminous tree Conzattia multiflora grown in Mexico.

    PubMed

    Wang, En Tao; Tan, Zhi Yuan; Guo, Xian Wu; Rodríguez-Duran, Rolando; Boll, Gisela; Martínez-Romero, Esperanza

    2006-10-01

    Conzattia multiflora is a leguminous tree present only in Mexico and Guatemala. There is no record about its symbiotic or pathogenic microbes. In this study, we found that numerous bacteria with 10(4)-10(6) individuals per gram of fresh epidermis were distributed in the tissue of this plant. All the bacteria isolated from the Conzattia epidermis were Gram-negative, facultative anaerobic rods and formed yellow or colorless colonies. They were identified as endophytes by inoculation tests. Some of the bacteria could significantly promote the growth of Conzattia seedlings. Nine different groups were defined by PCR-based RFLP, which were classified as Pantoea, Erwinia, Salmonella, Enterobacter, Citrobacter and Klebsiella by the phylogenetic analysis of 16S rRNA genes. The existence of plant-borne lineages of Salmonella indicates that the unexplored plants may harbor some unknown microbes.

  8. Methods for isolation of marine-derived endophytic fungi and their bioactive secondary products.

    PubMed

    Kjer, Julia; Debbab, Abdessamad; Aly, Amal H; Proksch, Peter

    2010-03-01

    Marine-derived fungi have been shown in recent years to produce a plethora of new bioactive secondary metabolites, some of them featuring new carbon frameworks hitherto unprecedented in nature. These compounds are of interest as new lead structures for medicine as well as for plant protection. The aim of this protocol is to give a detailed description of methods useful for the isolation and cultivation of fungi associated with various marine organisms (sponges, algae and mangrove plants) for the extraction, characterization and structure elucidation of biologically active secondary metabolites produced by these marine-derived endophytic fungi, and for the preliminary evaluation of their pharmacological properties based on rapid 'in house' screening systems. Some results exemplifying the positive outcomes of the protocol are given at the end. From sampling in marine environment to completion of the structure elucidation and bioactivity screening, a period of at least 3 months has to be scheduled.

  9. Endophytic bacteria isolated from orchid and their potential to promote plant growth.

    PubMed

    Faria, Deise Cristina; Dias, Armando Cavalcante Franco; Melo, Itamar Soares; de Carvalho Costa, Francisco Eduardo

    2013-02-01

    Twelve endophytic bacteria were isolated from the meristem of in vitro Cymbidium eburneum orchid, and screened according to indole yield quantified by colorimetric assay, in vitro phosphate solubilization, and potential for plant growth promotion under greenhouse conditions. Eight strains with positive results were classified into the genus Paenibacillus by FAME profile, and evaluated for their ability to increase survival and promote the growth of in vitro germinated Cattleya loddigesii seedlings during the acclimatization process. The obtained results showed that all strains produced detectable indole levels and did not exhibit potential for solubilizing inorganic phosphate. Particularly, an increase of the total biomass and number of leaves was observed. Two strains of Paenibacillus macerans promoted plant growth under greenhouse conditions. None of the treatments had a deleterious effect on growth of inoculated plants. These results suggest that these bacterial effects could be potentially useful to promote plant growth during seedling acclimatization in orchid species other than the species of origin.

  10. Geodermatophilus africanus sp. nov., a halotolerant actinomycete isolated from Saharan desert sand.

    PubMed

    Montero-Calasanz, María del Carmen; Göker, Markus; Pötter, Gabriele; Rohde, Manfred; Spröer, Cathrin; Schumann, Peter; Gorbushina, Anna A; Klenk, Hans-Peter

    2013-08-01

    A novel Gram-strain positive, aerobic, actinobacterial strain, designated CF11/1(T), was isolated from a sand sample obtained in the Sahara Desert, Chad. The black-pigmented isolate was aerobic and exhibited optimal growth from 25 to 35 °C at pH 6.0-8.0 and with 0-8 % (w/v) NaCl, indicating that it is a halotolerant mesophile. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The G+C content in the genome was 74.4 mol%. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and a minor fraction of phosphatidylglycerol; MK-9(H4) was the dominant menaquinone, and galactose was detected as a diagnostic sugar. The major cellular fatty acid was branched-chain saturated acid iso-C16:0. Analysis of 16S rRNA gene sequences showed 95.3-98.6 % pairwise sequence identity with the members of the genus Geodermatophilus. Based on phenotypic and chemotaxonomic properties, as well as phylogenetic distinctiveness, the isolate represents a novel species, Geodermatophilus africanus, with the type strain CF11/1(T) (DSM 45422 = CCUG 62969 = MTCC 11556).

  11. Actinomycetospora rhizophila sp. nov., an actinomycete isolated from rhizosphere soil of a peace lily (Spathi phyllum Kochii).

    PubMed

    He, Hairong; Zhang, Yuejing; Ma, Zhaoxu; Li, Chuang; Liu, Chongxi; Zhou, Ying; Li, Lianjie; Wang, Xiangjing; Xiang, Wensheng

    2015-05-01

    A novel actinomycete, designated strain NEAU-B-8(T), was isolated from the rhizosphere soil of a peace lily (Spathi phyllum Kochii) collected from Heilongjiang province, north-east China. Key morphological and physiological characteristics as well as chemotaxonomic features of strain NEAU-B-8(T) were congruent with the description of the genus Actinomycetospora , such as the major fatty acids, the whole-cell hydrolysates, the predominant menaquinone and the phospholipid profile. The 16S rRNA gene sequence analysis revealed that strain NEAU-B-8(T) shared the highest sequence similarities with Actinomycetospora lutea JCM 17982(T) (99.3% 16S rRNA gene sequence similarity), Actinomycetospora chlora TT07I-57(T) (98.4 %), Actinomycetospora straminea IY07-55(T) (98.3%) and Actinomycetospora chibensis TT04-21(T) (98.2%); similarities to type strains of other species of this genus were lower than 98%. The phylogenetic tree based on 16S rRNA gene sequences showed that strain NEAU-B-8(T) formed a distinct branch with A. lutea JCM 17982(T) that was supported by a high bootstrap value of 97% in the neighbour-joining tree and was also recovered with the maximum-likelihood algorithm. However, the DNA-DNA relatedness between strain NEAU-B-8(T) and A. lutea JCM 17982(T) was found to be 50.6 ± 1.2%. Meanwhile, strain NEAU-B-8(T) differs from other most closely related strains in phenotypic properties, such as maximum NaCl tolerance, hydrolysis of aesculin and decomposition of urea. On the basis of the morphological, physiological, chemotaxonomic, phylogenetic and DNA-DNA hybridization data, we conclude that strain NEAU-B-8(T) represents a novel species of the genus Actinomycetospora , named Actinomycetospora rhizophila sp. nov. The type strain is NEAU-B-8(T). ( = CGMCC 4.7134(T) =DSM 46673(T)).

  12. Actinokineospora soli sp. nov., a thermotolerant actinomycete isolated from soil, and emended description of the genus Actinokineospora.

    PubMed

    Tang, Xia; Zhou, Yu; Zhang, Jing; Ming, Hong; Nie, Guo-Xing; Yang, Ling-Ling; Tang, Shu-Kun; Li, Wen-Jun

    2012-08-01

    A strain of thermotolerant actinomycete, designated YIM 75948(T), was isolated from a soil sample in Yunnan province, China. The strain grew at 25-55 °C (optimum 37 °C). The substrate mycelium and aerial mycelium produced on Czapek's agar were both pale yellow to white. The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid, and the whole-cell sugars were mannose, ribose, glucose, galactose and arabinose. The major fatty acids were iso-C(15:0), iso-C(16:0) and C(16:0) and the predominant respiratory quinone was MK-9(H(4))(.) The polar lipids consisted of phosphatidylethanolamine, phosphatidylethanolamine with hydroxy fatty acids, diphosphatidylglycerol, phosphatidylinositol and two unidentified phospholipids. The genomic DNA G+C content was 73.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM 75948(T) belonged in the genus Actinokineospora and that its closest relative among recognized species was Actinokineospora fastidiosa DSM 43855(T) (97.6% sequence similarity). The mean level of DNA-DNA relatedness between the novel strain and A. fastidiosa DSM 43855(T) was, however, only 47.8%. Based on the phenotypic, chemotaxonomic and phylogenetic data and the results of the DNA-DNA hybridizations, strain YIM 75948(T) represents a novel species of the genus Actinokineospora for which the name Actinokineospora soli sp. nov. is proposed. The type strain is YIM 75948(T) ( =JCM 17695(T) =DSM 45613(T)). The description of the genus Actinokineospora is emended to reflect the fact that the genomic DNA G+C contents of A. fastidiosa DSM 43855(T) and the type strain of Actinokineospora soli sp. nov. recorded in the present study fell above the range given in previous descriptions of this genus.

  13. Fungal and plant gene expression during the colonization of cacao seedlings by endophytic isolates of four Trichoderma species.

    PubMed

    Bailey, B A; Bae, H; Strem, M D; Roberts, D P; Thomas, S E; Crozier, J; Samuels, G J; Choi, Ik-Young; Holmes, K A

    2006-11-01

    Endophytic isolates of Trichoderma species are being considered as biocontrol agents for diseases of Theobroma cacao (cacao). Gene expression was studied during the interaction between cacao seedlings and four endophytic Trichoderma isolates, T. ovalisporum-DIS 70a, T. hamatum-DIS 219b, T. harzianum-DIS 219f, and Trichoderma sp.-DIS 172ai. Isolates DIS 70a, DIS 219b, and DIS 219f were mycoparasitic on the pathogen Moniliophthora roreri, and DIS 172ai produced metabolites that inhibited growth of M. roreri in culture. ESTs (116) responsive to endophytic colonization of cacao were identified using differential display and their expression analyzed using macroarrays. Nineteen cacao ESTs and 17 Trichoderma ESTs were chosen for real-time quantitative PCR analysis. Seven cacao ESTs were induced during colonization by the Trichoderma isolates. These included putative genes for ornithine decarboxylase (P1), GST-like proteins (P4), zinc finger protein (P13), wound-induced protein (P26), EF-calcium-binding protein (P29), carbohydrate oxidase (P59), and an unknown protein (U4). Two plant ESTs, extensin-like protein (P12) and major intrinsic protein (P31), were repressed due to colonization. The plant gene expression profile was dependent on the Trichoderma isolate colonizing the cacao seedling. The fungal ESTs induced in colonized cacao seedlings also varied with the Trichoderma isolate used. The most highly induced fungal ESTs were putative glucosyl hydrolase family 2 (F3), glucosyl hydrolase family 7 (F7), serine protease (F11), and alcohol oxidase (F19). The pattern of altered gene expression suggests a complex system of genetic cross talk occurs between the cacao tree and Trichoderma isolates during the establishment of the endophytic association.

  14. Phylogenetic analysis of endophytic bacterial isolates from leaves of the medicinal plant Trichilia elegans A. Juss. (Meliaceae).

    PubMed

    Rhoden, S A; Garcia, A; Santos e Silva, M C; Azevedo, J L; Pamphile, J A

    2015-02-20

    Various organisms such as fungi and bacteria can live inside plants, inhabiting the aerial parts (primarily the leaves) without causing damage. These microorganisms, called endophytes, produce an extensive variety of compounds that can be useful for medical and agronomic purposes. Trichilia elegans A. Juss., belonging to the Meliaceae family, shows wide dispersion in South America, and phytochemical analyses from these plants and endophyte isolates have shown biological activity. Accordingly, the aim of this study was to verify the diversity of bacterial endophytes from T. elegans using partial sequencing of 16S rRNA, followed by phylogenetic analysis. Isolation was performed by cutting the leaves, after disinfection with 5% sodium hypochlorite (NaOCl), in 1-2-mm² fragments, which were equally placed on dishes containing TSA and fungicide BENLATE at 75 μg/mL. All dishes were incubated at 28°C in the biochemical oxygen demand system for 5 days and periodically checked. Afterwards, the colonization frequency (%) was determined: (number of fragments colonized by bacteria/total number of fragments) x 100. Three isolations between September 2011 and March 2012 were performed; the growth frequency ranged between 1.6 and 13.6%. Following sequencing of 16S rRNA and phylogenetic analysis, the genera identified were: Staphylococcus, Bacillus, Microbacterium, Pseudomonas, and Pantoea. These results will provide important knowledge on the diversity of endophytic bacteria inhabiting medicinal plants, and a better understanding of the microbiome of T. elegans would reinforce the necessity of endophyte studies with a focus on their future applications in biotechnological areas of agriculture, medicine, and the environment.

  15. Nocardiopsis oceani sp. nov. and Nocardiopsis nanhaiensis sp. nov., actinomycetes isolated from marine sediment of the South China Sea.

    PubMed

    Pan, Hua-Qi; Zhang, Dao-Feng; Li, Li; Jiang, Zhao; Cheng, Juan; Zhang, Yong-Guang; Wang, Hong-Fei; Hu, Jiang-Chun; Li, Wen-Jun

    2015-10-01

    Two actinomycete strains, designated 10A08AT and 10A08BT, were isolated from marine sediment samples of the South China Sea and their taxonomic positions were determined by a polyphasic approach. The two Gram-stain-positive, aerobic strains produced branched substrate mycelium and aerial hyphae, and no diffusible pigment was produced in the media tested. At maturity, spore chains were formed on aerial hyphae and all mycelium fragmented with age. Whole-cell hydrolysates of both strains contained meso-diaminopimelic acid and no diagnostic sugars. Their predominant menaquinones (>10 %) were MK-9(H4), MK-9(H6) and MK-10(H6) for strain 10A08AT and MK-9(H4), MK-9(H6), MK-10(H4) and MK-10(H6) for strain 10A08BT. The polar lipids detected from the two strains were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and unknown phosphoglycolipids and phospholipids. The major fatty acids (>10 %) of both strains were iso-C16 : 0 and summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B). The genomic DNA G+C contents of strains 10A08AT and 10A08BT were 70.9 and 71.6 mol%, respectively. On the basis of 16S rRNA gene sequence similarities, the two strains were shown to be most closely related to species of the genus Nocardiopsis. DNA–DNA hybridization relatedness values of < 70 % between these two isolates and their closest neighbour, Nocardiopsis terrae YIM 90022T, and between the two strains supported the conclusion that they represent two novel species. Based on phylogenetic analysis and phenotypic and genotypic data, it is concluded that the two isolates belong to the genus Nocardiopsis, and the names Nocardiopsis oceani sp. nov. (type strain 10A08AT = DSM 45931T = BCRC 16951T) and Nocardiopsis nanhaiensis sp. nov. (type strain 10A08BT = CGMCC 47227T = BCRC 16952T) are proposed.

  16. Isolation and characterization of actinomycetes strains that produce phospholipase D having high transphosphatidylation activity.

    PubMed

    Nakazawa, Yozo; Uchino, Masataka; Sagane, Yoshimasa; Sato, Hiroaki; Takano, Katsumi

    2009-01-01

    The present study was conducted to screen microorganisms that produce phospholipase D (PLD), and we especially focused on the strains having high transphosphatidylation activity. Eighty bacterial strains were isolated from soil samples by a screening method utilizing a preliminary selection medium with phosphatidylcholine (PC) as the sole carbon source. The culture supernatants were then assayed for PLD activity. The finding of dual PLD activities in cultures revealed that the hydrolytic and transphosphatidylation activities were correlated. Consequently, six strains were selected as stably producing PLD enzyme(s) during continuous subcultures. The culture supernatants of selected strains synthesized phosphatidylglycerol, phosphatidylserine and phosphatidylethanolamine from PC with high conversion rates. These isolated strains will be made available to carry out phospholipid modification through the efficient transphosphatidylation activity of the PLD that they produce.

  17. Geodermatophilus telluris sp. nov., an actinomycete isolated from Saharan desert sand.

    PubMed

    Montero-Calasanz, Maria del Carmen; Göker, Markus; Pötter, Gabriele; Rohde, Manfred; Spröer, Cathrin; Schumann, Peter; Klenk, Hans-Peter; Gorbushina, Anna A

    2013-06-01

    A novel Gram-positive, multiloculated thalli-forming, aerobic, actinobacterial strain, CF9/1/1(T), was isolated in 2007 during environmental screening for xerophilic fungi in arid desert soil from the Sahara desert, Chad. The isolate grew best at a temperature range of 20-35 °C and at pH 6.0-8.5 and with 0-4% (w/v) NaCl, forming black-coloured and irregular colonies on GYM agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G+C content of the novel strain was 75.4 mol%. The peptidoglycan contained meso-diaminopimelic acid as a diagnostic diamino acid. The main phospholipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, a not yet structurally identified aminophospholipid and a small amount of phosphatidylglycerol; MK-9(H4) was identified as the dominant menaquinone and galactose was a diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids: iso-C16:0 and iso-C15:0. The 16S rRNA gene sequence of the isolate showed 94.6-97.0% sequence similarities with those of five members of the genus: Geodermatophilus ruber DSM 45317(T) (94.6%), Geodermatophilus obscurus DSM 43160(T) (94.8%), Geodermatophilus siccatus DSM 45419(T) (96.2%), Geodermatophilus nigrescens DSM 45408(T) (96.7%) and Geodermatophilus arenarius DSM 45418(T) (97.0%). Based on the evidence from this polyphasic taxonomic study, a novel species, Geodermatophilus telluris sp. nov., is proposed; the type strain is CF9/1/1(T) (=DSM 45421(T)=CCUG 62764(T)).

  18. Geodermatophilus arenarius sp. nov., a xerophilic actinomycete isolated from Saharan desert sand in Chad.

    PubMed

    Montero-Calasanz, M C; Göker, M; Pötter, G; Rohde, M; Spröer, C; Schumann, P; Gorbushina, A A; Klenk, H-P

    2012-11-01

    A novel Gram-positive, aerobic, actinobacterial strain, CF5/4(T), was isolated in 2007 during an environmental screening of arid desert soil in Ouré Cassoni, Chad. The isolate grew best in a temperature range of 28-40 °C and at pH 6.0-8.5, with 0-1 % (w/v) NaCl, forming brown-coloured and nearly circular colonies on GYM agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content of the novel strain was 75.9 mol %. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, diphosphatidylglycerol and a small amount of phosphatidylglycerol; MK-9(H(4)) was identified as the dominant menaquinone and galactose as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids: iso-C(15:0) and iso-C(16:0). The 16S rRNA gene showed 96.2-98.3 % sequence identity with the three members of the genus Geodermatophilus: G. obscurus (96.2 %), G. ruber (96.5 %), and G. nigrescens (98.3 %). Based on the chemotaxonomic results, 16S rRNA gene sequence analysis and DNA-DNA hybridization with the type strain of G. nigrescens, the isolate is proposed to represent a novel species, Geodermatophilus arenarius (type strain CF5/4(T) = DSM 45418(T) = MTCC 11413(T) = CCUG 62763(T)).

  19. Fusaroside, a unique glycolipid from Fusarium sp., an endophytic fungus isolated from Melia azedarach.

    PubMed

    Yang, Sheng-Xiang; Wang, Hong-Peng; Gao, Jin-Ming; Zhang, Qiang; Laatsch, Hartmut; Kuang, Yi

    2012-01-28

    Fusaroside (1), a unique trehalose-containing glycolipid composed of the 4-hydroxyl group of a trehalose unit attached to the carboxylic carbon of a long-chain fatty acid, was isolated from the organic extract of fermentation broths of an endophytic fungus, Fusarium sp. LN-11 isolated from the leaves of Melia azedarach. Six known compounds, phalluside (2), (9R*,10R*,7E)-6, 9,10-trihydroxyoctadec-7-enoic acid (3), porrigenic acid (4), (9Z)-2,3-dihydroxypropyl octadeca-9-enoate (5), cerevisterol (6) and ergokonin B (7), were also isolated from this fungus. The glycolipid contains a rare branched long-chain fatty acid (C(20:4)) with a conjugated diene moiety and a conjugated ketone moiety. The structure of the new compound 1 was elucidated by spectroscopic methods (1D and 2D NMR experiments, MS) and chemical degradations. The metabolites 1-5 were shown to have moderate to weak active against the brine shrimp larvae. To our knowledge, this is the first report of isolation of the first representative of a new family of glycolipids from natural sources.

  20. Amycolatopsis rhabdoformis sp. nov., an actinomycete isolated from a tropical forest soil.

    PubMed

    Souza, Wallace Rafael; Silva, Rafael Eduardo; Goodfellow, Michael; Busarakam, Kanungnid; Figueiro, Fernanda Sales; Ferreira, Douglas; Rodrigues-Filho, Edson; Moraes, Luiz Alberto Beraldo; Zucchi, Tiago Domingues

    2015-06-01

    Strain SB026T was isolated from Brazilian rainforest soil and its taxonomic position established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological features consistent with its classification in the genus Amycolatopsis and formed a branch in the Amycolatopsis 16S rRNA gene tree together with Amycolatopsis bullii NRRL B-24847T, Amycolatopsis plumensis NRRL B-24324T, Amycolatopsis tolypomycina DSM 44544T and Amycolatopsis vancoresmycina NRRL B-24208T. It was related most closely to A. bullii NRRL B-24847T (99.0 % 16S rRNA gene sequence similarity), but was distinguished from this strain by a low level of DNA-DNA relatedness (~46 %) and discriminatory phenotypic properties. Based on the combined genotypic and phenotypic data, it is proposed that the isolate should be classified in the genus Amycolatopsis as representing a novel species, Amycolatopsis rhabdoformis sp. nov. The type strain is SB026T ( = CBMAI 1694T = CMAA 1285T = NCIMB 14900T).

  1. Streptomyces tritolerans sp. nov., a novel actinomycete isolated from soil in Karnataka, India.

    PubMed

    Syed, Dastager G; Agasar, Dayanand; Kim, Chang-Jin; Li, Wen-Jun; Lee, Jae-Chan; Park, Dong-Jin; Xu, Li-Hua; Tian, Xin-Peng; Jiang, Cheng-Lin

    2007-11-01

    A Gram-positive, nonmotile, moderately halophilic, alkali and thermotolerant strain designated DAS 165(T), was isolated from a dry land soil sample from the Gulbarga region, Karnataka province, India. The isolate produced yellow substrate mycelia and gray aerial mycelia on most tested media. Strain DAS 165(T) showed growth in the presence of 5 to 7% NaCl and at 45 degrees C. The DNA G + C content was 69.7%. 16S rRNA gene sequence analysis together with these characteristics consistently assigned strain DAS 165(T) to the genus Streptomyces. The 16S rRNA gene sequence analysis revealed that strain DAS 165(T) was most closely related to S. tendae ATCC 19812(T) (D 63873) with a sequence similarity of 99.6% (three nucleotide differences out of 1,517). Strain DAS 165(T) formed a distinct clade based on analysis of the almost complete sequence and 120-nucleotide variable gamma region of the 16S rRNA gene. Despite the high sequence similarity, strain DAS 165(T) was phenotypically different from S. tendae ATCC 19812(T). DNA-DNA hybridization between these strains was 47% showing that strain DAS 165(T) is a distinct genomic species. Phenetic and genetic results support the classification of strain DAS 165(T) as a new species, for which the name S. tritolerans is proposed, with strain DAS 165(T) as the type strain (=DSM 41899(T )= CCTCCAA 206013(T)).

  2. Verrucosispora fiedleri sp. nov., an actinomycete isolated from a fjord sediment which synthesizes proximicins.

    PubMed

    Goodfellow, Michael; Brown, Roselyn; Ahmed, Lina; Pathom-Aree, Wasu; Bull, Alan T; Jones, Amanda L; Stach, James E M; Zucchi, Tiago Domingues; Zhang, Lixin; Wang, Jian

    2013-03-01

    A novel filamentous actinobacterial organism, designated strain MG-37(T), was isolated from a Norwegian fjord sediment and examined using a polyphasic taxonomic approach. The organism was determined to have chemotaxonomic and morphological properties consistent with its classification in the genus Verrucosispora and formed a distinct phyletic line in the Verrucosispora 16S rRNA gene tree. It was most closely related to Verrucosispora maris DSM 45365(T) (99.5 % 16S rRNA gene similarity) and Verrucosispora gifhornensis DSM 44337(T) (99.4 % 16S rRNA gene similarity) but was distinguished from these strains based on low levels of DNA:DNA relatedness (~56 and ~50 %, respectively). It was readily delineated from all of the type strains of Verrucosispora species based on a combination of phenotypic properties. Isolate MG-37(T) (=NCIMB 14794(T) = NRRL-B-24892(T)) should therefore be classified as the type strain of a novel species of Verrucosispora for which the name Verrucosispora fiedleri is proposed.

  3. Morphological, physiological, and molecular characterization of actinomycetes isolated from dry soil, rocks, and monument surfaces.

    PubMed

    Eppard, M; Krumbein, W E; Koch, C; Rhiel, E; Staley, J T; Stackebrandt, E

    1996-07-01

    In an extended study on the biodiversity of rock-dwelling bacteria, the colony and cell morphology, physiology, protein patterns, and 16S rDNA sequences of 17 bacterial strains isolated from different surfaces of rocks, stones, and monuments and from various geographical locations were characterized. All except one strain, which was found to be a Bacillus, were members of the order Actinomycetales. The majority of the strains either were closely related to Geodermatophilus obscurus, which was also analyzed in this study, or formed a closely related sister taxon. All of these strains were isolated from the surface of marble in Namibia and Greece and from limestone from the Negev desert, Israel. One strain, G10, of Namibia origin was equidistantly related to Geodermatophilus obscurus, Frankia alni, Sporichthya polymorpha, and Acidothermus cellulolyticus. Three strains from rock varnish in the Mojave desert, California, were found to be highly related to Arthrobacter (formerly Micrococcus) agilis. All clusters could be confirmed from results of studies on morphological and physiological properties and from banding patterns of whole cell proteins. Based on the results of tests, four additional strains were assigned to the lineage defined by strain G10.

  4. Beutenbergia cavernae gen. nov., sp. nov., an L-lysine-containing actinomycete isolated from a cave.

    PubMed

    Groth, I; Schumann, P; Schuetze, B; Augsten, K; Kramer, I; Stackebrandt, E

    1999-10-01

    Two aerobic, Gram-positive bacteria, strains HKI 0122T and HKI 0132, were isolated from a cave. Cells are not acid-fast, non-motile, non-spore-forming and exhibit a rod-coccus growth cycle. The cell wall peptidoglycan contains lysine in position 3 of the peptide subunit and an interpeptide bridge of L-Lys<--L-Glu. The major menaquinone is MK-8(H4), 13-methyl and 12-methyl tetradecanoic acids are the predominating fatty acids. The polar lipids consist of phosphatidylinositol, diphosphatidylglycerol and three unknown phospholipids. Mycolic acids are absent. The DNA base composition is 71 mol% G + C. Phylogenetic analysis revealed that strain HKI 0122T forms a novel taxon among the families and unassigned genera of the suborder Micrococcineae, within the order Actinomycetales. On the basis of the genotypic, chemotaxonomic, morphological and physiological characteristics of these two isolates it is proposed to assign strains HKI 0122T and HKI 0132 to a new genus and species for which the name Beutenbergia cavernae gen. nov., sp. nov. is proposed. The type strain is HKI 0122T (= DSM 12333T).

  5. Potential of Endophytic Fungi Isolated from Cotton Roots for Biological Control against Verticillium Wilt Disease.

    PubMed

    Yuan, Yuan; Feng, Hongjie; Wang, Lingfei; Li, Zhifang; Shi, Yongqiang; Zhao, LiHong; Feng, Zili; Zhu, Heqin

    2017-01-01

    Verticillium wilt is a soil-borne disease, and severely limits the development of cotton production. To investigate the role of endophytic fungi on Verticillium wilt, CEF-818 (Penicillium simplicissimum), CEF-714 (Leptosphaeria sp.), CEF-642 (Talaromyces flavus.) and CEF-193 (Acremonium sp.) isolated from cotton roots were used to assess their effects against cotton wilt disease caused by a defoliating V. dahliae strain Vd080. In the greenhouse, all treatments significantly reduced disease incidence and disease index, with the control efficacy ranging from 26% (CEF-642) to 67% (CEF-818) at 25 days (d) after inoculation. In the disease nursery, compared to controls (with disease incidence of 33.8% and disease index of 31), CEF-818, CEF-193, CEF-714 and CEF-642 provided a protection effect of 69.5%, 69.2%, 54.6% and 45.7%, respectively. Especially, CEF-818 and CEF-714 still provided well protection against Verticillium wilt with 46.9% and 56.6% or 14.3% and 33.7% at the first peak of the disease in heavily infected field, respectively (in early July). These results indicated that these endophytes not only delayed but also reduced wilt symptoms on cotton. In the harvest, the available cotton bolls of plant treated with CEF-818 and CEF-714 increased to 13.1, and 12.2, respectively. And the seed cotton yield significantly increased after seed bacterization with CEF-818 (3442.04 kg/ha) compared to untreated control (3207.51 kg/ha) by 7.3%. Furtherly, CEF-818 and CET-714 treatment increased transcript levels for PAL, PPO, POD, which leads to the increase of cotton defense reactions. Our results indicate that seed treatment of cotton plants with CEF-818 and CET-714 can help in the biocontrol of V. dahliae and improve seed cotton yield in cotton fields. This study provided a better understanding of cotton-endophyte interactions which will aid in developing effective biocontrol agents for Verticillium wilt of cotton in futhre.

  6. Potential of Endophytic Fungi Isolated from Cotton Roots for Biological Control against Verticillium Wilt Disease

    PubMed Central

    Wang, Lingfei; Li, Zhifang; Shi, Yongqiang; Zhao, LiHong; Feng, Zili; Zhu, Heqin

    2017-01-01

    Verticillium wilt is a soil-borne disease, and severely limits the development of cotton production. To investigate the role of endophytic fungi on Verticillium wilt, CEF-818 (Penicillium simplicissimum), CEF-714 (Leptosphaeria sp.), CEF-642 (Talaromyces flavus.) and CEF-193 (Acremonium sp.) isolated from cotton roots were used to assess their effects against cotton wilt disease caused by a defoliating V. dahliae strain Vd080. In the greenhouse, all treatments significantly reduced disease incidence and disease index, with the control efficacy ranging from 26% (CEF-642) to 67% (CEF-818) at 25 days (d) after inoculation. In the disease nursery, compared to controls (with disease incidence of 33.8% and disease index of 31), CEF-818, CEF-193, CEF-714 and CEF-642 provided a protection effect of 69.5%, 69.2%, 54.6% and 45.7%, respectively. Especially, CEF-818 and CEF-714 still provided well protection against Verticillium wilt with 46.9% and 56.6% or 14.3% and 33.7% at the first peak of the disease in heavily infected field, respectively (in early July). These results indicated that these endophytes not only delayed but also reduced wilt symptoms on cotton. In the harvest, the available cotton bolls of plant treated with CEF-818 and CEF-714 increased to 13.1, and 12.2, respectively. And the seed cotton yield significantly increased after seed bacterization with CEF-818 (3442.04 kg/ha) compared to untreated control (3207.51 kg/ha) by 7.3%. Furtherly, CEF-818 and CET-714 treatment increased transcript levels for PAL, PPO, POD, which leads to the increase of cotton defense reactions. Our results indicate that seed treatment of cotton plants with CEF-818 and CET-714 can help in the biocontrol of V. dahliae and improve seed cotton yield in cotton fields. This study provided a better understanding of cotton-endophyte interactions which will aid in developing effective biocontrol agents for Verticillium wilt of cotton in futhre. PMID:28107448

  7. Rational approaches to improving the isolation of endophytic actinobacteria from Australian native trees.

    PubMed

    Kaewkla, Onuma; Franco, Christopher M M

    2013-02-01

    In recent years, new actinobacterial species have been isolated as endophytes of plants and shrubs and are sought after both for their role as potential producers of new drug candidates for the pharmaceutical industry and as biocontrol inoculants for sustainable agriculture. Molecular-based approaches to the study of microbial ecology generally reveal a broader microbial diversity than can be obtained by cultivation methods. This study aimed to improve the success of isolating individual members of the actinobacterial population as pure cultures as well as improving the ability to characterise the large numbers obtained in pure culture. To achieve this objective, our study successfully employed rational and holistic approaches including the use of isolation media with low concentrations of nutrients normally available to the microorganism in the plant, plating larger quantities of plant sample, incubating isolation plates for up to 16 weeks, excising colonies when they are visible and choosing Australian endemic trees as the source of the actinobacteria. A hierarchy of polyphasic methods based on culture morphology, amplified 16S rRNA gene restriction analysis and limited sequencing was used to classify all 576 actinobacterial isolates from leaf, stem and root samples of two eucalypts: a Grey Box and Red Gum, a native apricot tree and a native pine tree. The classification revealed that, in addition to 413 Streptomyces spp., isolates belonged to 16 other actinobacterial genera: Actinomadura (two strains), Actinomycetospora (six), Actinopolymorpha (two), Amycolatopsis (six), Gordonia (one), Kribbella (25), Micromonospora (six), Nocardia (ten), Nocardioides (11), Nocardiopsis (one), Nonomuraea (one), Polymorphospora (two), Promicromonospora (51), Pseudonocardia (36), Williamsia (two) and a novel genus Flindersiella (one). In order to prove novelty, 12 strains were characterised fully to the species level based on polyphasic taxonomy. One strain represented a novel

  8. Streptomyces pini sp. nov., an actinomycete isolated from phylloplane of pine (Pinus sylvestris L.) needle-like leaves.

    PubMed

    Madhaiyan, Munusamy; Poonguzhali, Selvaraj; Saravanan, Venkatakrishnan Sivaraj; Duraipandiyan, Veeramuthu; Al-Dhabi, Naif Abdullah; Pragatheswari, Dhandapani; Santhanakrishnan, Palani; Kim, Soo-Jin; Weon, Hang-Yeon; Kwon, Soon-Wo

    2016-10-01

    A novel siderophore-producing actinomycete, designated PL19T, was isolated from the Scots-pine needle-like leaves collected from TNAU campus, Coimbatore, India. The isolate was chemoorganotrophic in nutrition and able to grow at 30 °C, and the optimum pH and NaCl facilitated the growth pH 6-11 and 0-8 % (w/v), respectively. The cells are filamentous and the mycelia formed are basically of wide and intricately branched substrate mycelium from which aerial mycelia arises, later gets differentiated into spores that are warty and arranged spirally. The 16S rRNA gene of strain PL19T was sequenced and was highly similar to the type strains of species of the genus Streptomyces, including Streptomyces barkulensis RC1831T (98.8 % pairwise similarity), Streptomyces fenghuangensis GIMN4.003T (98.2 %), Streptomyces nanhaiensis SCSIO 01248T (98.0 %), Streptomyces radiopugnans R97T (97.9 %), Streptomyces atacamensis C60T (97.8 %) and Streptomyces macrosporus NBRC 14749T (97.2 %), all of which were subjected to taxonomical characterization using a polyphasic approach. The strains showed unique carbon utilization patterns, and it possesses iso-C16 : 0 anteiso-C15 : 0 and anteiso-C17 : 0 as a major cellular fatty acids. The cell-wall was dominated with ll-type diaminopimelic acid, and the menaquinone type was MK-9(H6, H8). These chemotaxonomic evidences placed strain PL19T within the genus Streptomyces. The determination of G+C ratio (69.5 mol%) and DNA-DNA hybridization values (13.4-31.8 % with the phylogenetically related species) helped in further hierarchical classification of strain PL19T. Based on morphological, physiological and chemotaxonomic data as well as DNA-DNA hybridization values, strain PL19T could be distinguished from the evolutionarily closest species currently available. All these collective data show that strain PL19T represents a novel species of the genus Streptomyces, for which the name Streptomyces pini sp. nov. is proposed

  9. Geodermatophilus poikilotrophi sp. nov.: a multitolerant actinomycete isolated from dolomitic marble.

    PubMed

    del Carmen Montero-Calasanz, Maria; Hofner, Benjamin; Göker, Markus; Rohde, Manfred; Spröer, Cathrin; Hezbri, Karima; Gtari, Maher; Schumann, Peter; Klenk, Hans-Peter

    2014-01-01

    A novel Gram-reaction-positive, aerobic actinobacterium, tolerant to mitomycin C, heavy metals, metalloids, hydrogen peroxide, desiccation, and ionizing- and UV-radiation, designated G18T, was isolated from dolomitic marble collected from outcrops in Samara (Namibia). The growth range was 15-35°C, at pH 5.5-9.5 and in presence of 1% NaCl, forming greenish-black coloured colonies on GYM Streptomyces agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for other representatives of the genus Geodermatophilus. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, and small amount of diphosphatidylglycerol. MK-9(H4) was the dominant menaquinone and galactose was detected as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids iso-C16:0 and iso-C15:0 and the unsaturated C17:1 ω8c and C16:1 ω7c. The 16S rRNA gene showed 97.4-99.1% sequence identity with the other representatives of genus Geodermatophilus. Based on phenotypic results and 16S rRNA gene sequence analysis, strain G18T is proposed to represent a novel species, Geodermatophilus poikilotrophi. Type strain is G18T (=DSM 44209T=CCUG 63018T). The INSDC accession number is HF970583. The novel R software package lethal was used to compute the lethal doses with confidence intervals resulting from tolerance experiments.

  10. Geodermatophilus poikilotrophi sp. nov.: A Multitolerant Actinomycete Isolated from Dolomitic Marble

    PubMed Central

    Montero-Calasanz, Maria del Carmen; Hofner, Benjamin; Göker, Markus; Rohde, Manfred; Spröer, Cathrin; Hezbri, Karima; Gtari, Maher; Schumann, Peter; Klenk, Hans-Peter

    2014-01-01

    A novel Gram-reaction-positive, aerobic actinobacterium, tolerant to mitomycin C, heavy metals, metalloids, hydrogen peroxide, desiccation, and ionizing- and UV-radiation, designated G18T, was isolated from dolomitic marble collected from outcrops in Samara (Namibia). The growth range was 15–35°C, at pH 5.5–9.5 and in presence of 1% NaCl, forming greenish-black coloured colonies on GYM Streptomyces agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for other representatives of the genus Geodermatophilus. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, and small amount of diphosphatidylglycerol. MK-9(H4) was the dominant menaquinone and galactose was detected as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids iso-C16:0 and iso-C15:0 and the unsaturated C17:1 ω8c and C16:1 ω7c. The 16S rRNA gene showed 97.4–99.1% sequence identity with the other representatives of genus Geodermatophilus. Based on phenotypic results and 16S rRNA gene sequence analysis, strain G18T is proposed to represent a novel species, Geodermatophilus poikilotrophi. Type strain is G18T (= DSM 44209T = CCUG 63018T). The INSDC accession number is HF970583. The novel R software package lethal was used to compute the lethal doses with confidence intervals resulting from tolerance experiments. PMID:25114928

  11. Streptomyces manipurensis sp. nov., a novel actinomycete isolated from a limestone deposit site in Manipur, India.

    PubMed

    Nimaichand, Salam; Zhu, Wen-Yong; Yang, Ling-Ling; Ming, Hong; Nie, Guo-Xing; Tang, Shu-Kun; Ningthoujam, Debananda S; Li, Wen-Jun

    2012-06-01

    A novel actinobacterium, designated MBRL 201(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. The strain was characterized using polyphasic taxonomy. Comparison of the 16S rRNA gene sequence of strain MBRL 201(T) and other Streptomyces species showed sequence similarities ranging from 93.0 to 99.6 % and strain MBRL 201(T) showed closest similarities to Streptomyces virginiae NBRC 12827(T) (99.6 %) and Streptomyces cinnamonensis NBRC 15873(T) (99.6 %). The DNA relatedness between MBRL 201(T) and the type strains of S. virginiae NBRC 12827(T) and S. cinnamonensis NBRC 15873(T) were 44.5 and 35.6 % respectively. Strain MBRL 201(T) contained LL: -diaminopimelic acid (A(2)pm) as the diagnostic diamino acid, with glucose as the main sugar, while small amounts of galactose, glucose, mannose, rhamnose, ribose and xylose were also present in cell-wall hydrolysates. The major fatty acids identified were anteiso-C(15:0) (38.9 %), iso-C(15:0) (19.9 %) and anteiso-C(17:1) (14.7 %). The predominant menaquinones detected were MK-9(H(6)) and MK-9(H(8)), while the polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides, with other unknown phospholipids and lipids. The G+C content of the genomic DNA was 72.9 %. The phenotypic and genotypic data showed that strain MBRL 201(T) merits recognition as a representative of a novel species of the genus Streptomyces. It is proposed that the isolate should be classified in the genus Streptomyces as a novel species, Streptomyces manipurensis sp. nov. The type strain is MBRL 201(T) (=DSM 42029(T) = JCM 17351(T)).

  12. Description of Actinokineospora acnipugnans sp. nov., an actinomycete isolated from soil, showing potential uses in cosmetics.

    PubMed

    Dahal, Ram Hari; Shim, Dong Seop; Kim, Jaisoo

    2017-08-01

    During isolation of soil bacteria having antibacterial functions, an aerobic, Gram-stain-positive, oxidase-negative, catalase-positive bacterium, designated strain R434T, was isolated. Strain R434T showed antimicrobial activity against Propionibacterium acnes and Staphylococcus epidermidis and significant enzyme-inhibitory capability. The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid, and the whole-cell sugars were galactose, arabinose and glucose. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain R434T formed a lineage within the family Pseudonocardiaceae. Strain R434T showed highest sequence similarity with type strains of the genus Actinokineospora, including Actinokineospora guangxiensis Gk-6T (99.4 % sequence similarity), Actinokineospora soli YIM 75948T (98.5 %), Actinokineospora fastidiosa IMSNU 20054T (98.0 %), Actinokineospora cibodasensis ID03-0784T (97.9 %), Actinokineospora terrae IFO 15668T (97.6 %) and Actinokineospora auranticolor IFO 16518T (97.4 %). The predominant respiratory quinone of strain R434T was MK-9(H4). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine and unidentified glycolipid. The major cellular fatty acids were iso-C16 : 0, iso-C16 : 1 H and C17 : 1ω6c. The DNA G+C content of strain R434T was 71.6 mol%. On the basis of phenotypic, genotypic, chemotaxonomic and phylogenetic analysis, strain R434T represents a novel species of the genus Actinokineospora, for which the name Actinokineospora acnipugnans sp. nov. is proposed. The type strain of 'Actinokineospora acnipugnans' is R434T (=KEMB 9005-403T=KACC 18904T=JCM 31557T).

  13. Rhodococcus sovatensis sp. nov., an actinomycete isolated from the hypersaline and heliothermal Lake Ursu.

    PubMed

    Táncsics, András; Máthé, István; Benedek, Tibor; Tóth, Erika M; Atasayar, Ewelina; Spröer, Cathrin; Márialigeti, Károly; Felföldi, Tamás; Kriszt, Balázs

    2017-02-01

    A Gram-stain-positive, strictly aerobic, mesophilic bacterium, designated H004T, was isolated from a water sample of the hypersaline and heliothermal Lake Ursu, Sovata, Romania. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain H004T formed a distinct phyletic lineage within the genus Rhodococcus. It shared the highest 16S rRNA gene sequence similarity with Rhodococcus yunnanensis YIM 70056T (98.80 %), followed by Rhodococcus fascians LMG 3623T (98.73 %), Rhodococcus cercidiphylli YIM 65003T (98.73 %), Rhodococcus cerastii C5T (98.58 %) and Rhodococcus kyotonensis DS472T (98.53 %). The alkB-based phylogenetic analysis further confirmed that this strain constitutes a highly unique lineage within the genus. Chemotaxonomic characteristics, including the predominant fatty acids acids C15 : 0, C18 : 1ω9c, C19 : 1ω11c/C19 : 1ω9c and C16 : 1ω7c/iso-C15 : 0 2-OH, the major quinone MK-8(H2), the presence of mycolic acids and cell-wall chemotype IV were also consistent with the properties of members of the genus Rhodococcus. The DNA G+C content of strain H004T was 65.4 mol%. The results of DNA-DNA hybridization analyses with the closest relatives, in combination with the alkB-based phylogenetic analysis, as well as the chemotaxonomic and physiological data, demonstrated that isolate H004T represents a novel species of the genus Rhodococcus, for which the name Rhodococcus sovatensissp. nov. is proposed. The type strain is H004T (=DSM 102881T=NCAIM B.02632T).

  14. Fodinicola feengrottensis gen. nov., sp. nov., an actinomycete isolated from a medieval mine.

    PubMed

    Carlsohn, Marc René; Groth, Ingrid; Saluz, Hans-Peter; Schumann, Peter; Stackebrandt, Erko

    2008-07-01

    A filamentous, Gram-positive actinobacterium was isolated from acidic rocks in a medieval alum slate mine and was investigated by means of a polyphasic taxonomic approach. A 16S rRNA gene sequence similarity study indicated that strain HKI 0501(T) forms an individual line of descent and is related to certain members of the suborder Frankineae, order Actinomycetales (<95 % sequence similarity). Distance-matrix and neighbour-joining analyses set the branching point of the novel isolate between two clades, one being represented by members of the genus Cryptosporangium (family 'Kineosporiaceae') and the other by members of the genera Frankia and Acidothermus (family Frankiaceae and family Acidothermaceae, respectively). The organism had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan and xylose as the characteristic cell-wall sugar. The muramic acid in the peptidoglycan was found to be N-acetylated. The major menaquinones were MK-9(H(4)), MK-9(H(6)) and MK-9(H(8)) and the fatty acid profile was characterized by the predominance of iso-C(16 : 0), 10-methyl C(17 : 0), C(17 : 1) cis9 and 10-methyl iso-C(18 : 0). The polar lipids comprised diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and several unknown phospholipids and glycolipids. Mycolic acids were absent. The DNA G+C content was 65 mol%. The distinct phylogenetic position and the phenotypic markers that clearly separate the novel organism from all other members of the suborder Frankineae indicate that strain HKI 0501(T) represents a novel genus and species, for which the name Fodinicola feengrottensis gen. nov., sp. nov. is proposed. The type strain of Fodinicola feengrottensis is HKI 0501(T) (=DSM 19247(T) =JCM 14718(T)).

  15. Streptomyces xinjiangensis sp. nov., an actinomycete isolated from Lop Nur region.

    PubMed

    Cheng, Cong; Li, Yu-Qian; Asem, Mipeshwaree Devi; Lu, Chun-Yan; Shi, Xiao-Han; Chu, Xiao; Zhang, Wan-Qin; Di An, Deng-; Li, Wen-Jun

    2016-10-01

    A novel actinobacterial strain, designated LPA192(T), was isolated from a soil sample collected from Lop Nur, Xinjiang Uygur Autonomous Region, Northwest China. A polyphasic approach was used to investigate the taxonomic position of strain LPA192(T). The isolate showed morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Peptidoglycan was found to contain LL-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinones were MK-9(H6) and MK-10(H4). Polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol. Major cellular fatty acids consist of C16:0, anteiso-C15:0 and C18:1 ω9c. The sugar in whole-cell hydrolysates was mannose. Phylogenetic analysis indicated that strain LPA192(T) is closely related to Streptomyces tanashiensis LMG 20274(T) (99.3 %), Streptomyces gulbargensis DAS131(T) (99.3 %), Streptomyces nashvillensis NBRC 13064(T) (99.3 %), Streptomyces roseolus NBRC 12816(T) (99.2 %) and Streptomyces filamentosus NBRC 12767(T) (99.1 %) while showing below 98.5 % sequencing similarities with other validly published Streptomyces species. However, DNA-DNA relatedness values between LPA192(T) and the closely related type strains were below 40 %, which are much lower than 70 % threshold value for species delineation. The genomic DNA G + C content of strain LPA192(T) was 69.3 mol %. Based on the differences in genotypic and phenotypic characteristics from the closely related strains, strain LPA192(T) is considered to represent a novel species of the genus Streptomyces for which the name Streptomyces xinjiangensis sp. nov. is proposed. The type strain is LPA192(T) (=KCTC 39601(T) = CGMCC 4.7288(T)).

  16. A Comparative biochemical study on two marine endophytes, Bacterium SRCnm and Bacillus sp. JS, Isolated from red sea algae.

    PubMed

    Ahmed, Eman Fadl; Hassan, Hossam Mokhtar; Rateb, Mostafa Ezzat; Abdel-Wahab, Noha; Sameer, Somayah; Aly Taie, Hanan Anwar; Abdel-Hameed, Mohammed Sayed; Hammouda, Ola

    2016-01-01

    Two marine endophytic bacteria were isolated from the Red Sea algae; a red alga; Acanthophora dendroides and the brown alga Sargassum sabrepandum. The isolates were identified based on their 16SrRNA sequences as Bacterium SRCnm and Bacillus sp. JS. The objective of this study was to investigate the potential anti-microbial and antioxidant activities of the extracts of the isolated bacteria grown in different nutrient conditions. Compared to amoxicillin (25μg/disk) and erythromycin (15μg/disk), the extracts of Bacterium SRCn min media II, III, IV and V were potent inhibitors of the gram-positive bacterium Sarcina maxima even at low concentrations. Also, the multidrug resistant Staphylococcus aureus(MRSA) was more sensitive to the metabolites produced in medium (II) of the same endophyte than erythromycin (15μg/disk). A moderate activity of the Bacillus sp. JS extracts of media I and II was obtained against the same pathogen. The total compounds (500ug/ml) of both isolated endophytes showed moderate antioxidant activities (48.9% and 46.1%, respectively). LC/MS analysis of the bacterial extracts was carried out to investigate the likely natural products produced. Cyclo(D-cis-Hyp-L-Leu), dihydrosphingosine and 2-Amino-1,3-hexadecanediol were identified in the fermentation medium of Bacterium SRCnm, whereas cyclo (D-Pro-L-Tyr) and cyclo (L-Leu-L-Pro) were the suggested compounds of Bacillus sp. JS.

  17. Enrichment culture and identification of endophytic methanotrophs isolated from peatland plants.

    PubMed

    Stępniewska, Zofia; Goraj, Weronika; Kuźniar, Agnieszka; Łopacka, Natalia; Małysza, Magdalena

    2017-03-09

    Aerobic methane-oxidizing bacteria (MOB) are an environmentally significant group of microorganisms due to their role in the global carbon cycle. Research conducted over the past few decades has increased the interest in discovering novel genera of methane-degrading bacteria, which efficiently utilize methane and decrease the global warming effect. Moreover, methanotrophs have more promising applications in environmental bioengineering, biotechnology, and pharmacy. The investigations were undertaken to recognize the variety of endophytic methanotrophic bacteria associated with Carex nigra, Vaccinium oxycoccus, and Eriophorum vaginatum originating from Moszne peatland (East Poland). Methanotrophic bacteria were isolated from plants by adding sterile fragments of different parts of plants (roots and stems) to agar mineral medium (nitrate mineral salts (NMS)) and incubated at different methane values (1-20% CH4). Single colonies were streaked on new NMS agar media and, after incubation, transferred to liquid NMS medium. Bacterial growth dynamics in the culture solution was studied by optical density-OD600 and methane consumption. Changes in the methane concentration during incubation were controlled by the gas chromatography technique. Characterization of methanotrophs was made by fluorescence in situ hybridization (FISH) with Mg705 and Mg84 for type I methanotrophs and Ma450 for type II methanotrophs. Identification of endophytes was performed after 16S ribosomal RNA (rRNA) and mmoX gene amplification. Our study confirmed the presence of both types of methanotrophic bacteria (types I and II) with the predominance of type I methanotrophs. Among cultivable methanotrophs, there were different strains of the genus Methylomonas and Methylosinus. Furthermore, we determined the potential of the examined bacteria for methane oxidation, which ranged from 0.463 ± 0.067 to 5.928 ± 0.169 μmol/L CH4/mL/day.

  18. Rhodococcus cercidiphylli sp. nov., a new endophytic actinobacterium isolated from a Cercidiphyllum japonicum leaf.

    PubMed

    Li, Jie; Zhao, Guo-Zhen; Chen, Hua-Hong; Qin, Sheng; Xu, Li-Hua; Jiang, Cheng-Lin; Li, Wen-Jun

    2008-06-01

    An endophytic actinobacterium, designated YIM 65003(T), was isolated from a surface sterilized leaf sample of Cercidiphyllum japonicum collected from Yunnan province, south-west China. The morphological and chemotaxonomic properties of the isolate were typical of members of the genus Rhodococcus. Analysis of the 16S rRNA gene sequence revealed that the isolate was most closely related to Rhodococcus fascians DSM 20669(T) (99.6%) and Rhodococcus yunnanensis YIM 70056(T) (99.0%). DNA-DNA hybridization with the above microorganisms (46.3% and 48.8%, respectively), in combination with differences in the biochemical and physiological properties, suggested that strain YIM 65003(T) should be classified within a novel species of the genus Rhodococcus, for which the name Rhodococcus cercidiphylli sp. nov. is proposed, with YIM 65003(T) (=CCTCC AB 207160(T)=DSM 45141(T)) as the type strain. The 16S rRNA gene sequence of strain YIM 65003(T) has been deposited in GenBank under the accession number EU325542.

  19. Grass fungal endophytes and uses thereof

    DOEpatents

    Craven, Kelly

    2015-03-10

    The invention provides isolated fungal endophytes and synthetic combinations thereof with host grass plants. Methods for inoculating grass plant with the endophytes, for propagating the grass-endophyte combinations, and for producing feeds and biofuels from grass-endophyte combinations are also provided.

  20. Tsukamurella spongiae sp. nov., a novel actinomycete isolated from a deep-water marine sponge.

    PubMed

    Olson, Julie B; Harmody, Dedra K; Bej, Asim K; McCarthy, Peter J

    2007-07-01

    A Gram-positive, rod-shaped, non-spore-forming bacterium (strain K362(T)) was isolated from a deep-water marine sponge collected off the coast of Curaçao in the Netherlands Antilles. On the basis of 16S rRNA gene sequence similarities, strain K362(T) was shown to belong to the genus Tsukamurella, being most closely related to Tsukamurella pulmonis (99.2 %), Tsukamurella tyrosinosolvens (98.9 %), Tsukamurella strandjordii (98.8 %), Tsukamurella pseudospumae (98.8 %) and Tsukamurella spumae (98.8 %). A combination of the substrate utilization patterns, the fatty acid and mycolic acid profiles and the DNA-DNA hybridization results supported the affiliation of strain K362(T) to the genus Tsukamurella and enabled the genotypic and phenotypic differentiation of strain K362(T) from the seven recognized Tsukamurella species. Strain K362(T) therefore represents a novel species of the genus Tsukamurella, for which the name Tsukamurella spongiae sp. nov. is proposed. The type strain is K362(T) (=DSM 44990(T)=NRRL B-24467(T)).

  1. Streptomyces songpinggouensis sp. nov., a Novel Actinomycete Isolated from Soil in Sichuan, China.

    PubMed

    Guan, Xuejiao; Li, Wenchao; Liu, Chongxi; Jin, Pinjiao; Guo, Siyu; Wang, Xiangjing; Xiang, Wensheng

    2016-12-01

    During a screening for novel and biotechnologically useful actinobacteria, a novel actinobacteria with weak antifungal activity, designated strain NEAU-Spg19(T), was isolated from a soil sample collected from pine forest in Songpinggou, Sichuan, southwest China. The strain was characterized using a polyphasic taxonomic approach which confirmed that it belongs to the genus Streptomyces. Growth occurred at a temperature range of 10-30 °C, pH 5.0-11.0 and NaCl concentrations of 0-5 %. The cell wall peptidoglycan consisted of LL-diaminopimelic acid and glycine. The major menaquinones were MK-9(H6), MK-9(H8) and MK-9(H4). The phospholipid profile contained diphosphatidylglycerol (DPG), phosphatidylethanolamine and phosphatidylinositol. The major fatty acids were iso-C15:0, iso-C16:0, and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-Spg19(T) belongs to the genus Streptomyces with the highest sequence similarities to Streptomyces tauricus JCM 4837(T) (98.6 %) and Streptomyces rectiviolaceus JCM 9092(T) (98.3 %). Some physiological and biochemical properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. tauricus JCM 4837(T) and S. rectiviolaceus JCM 9092(T). Hence, on the basis of phenotypic and genetic analyses, it is proposed that strain NEAU-Spg19(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces songpinggouensis sp. nov. is proposed. The type strain is NEAU-Spg19(T) (=CGMCC 4.7140(T)=DSM 42141(T)).

  2. Rhodococcus canchipurensis sp. nov., an actinomycete isolated from a limestone deposit site.

    PubMed

    Nimaichand, Salam; Sanasam, Suchitra; Zheng, Liu-Qiang; Zhu, Wen-Yong; Yang, Ling-Ling; Tang, Shu-Kun; Ningthoujam, Debananda S; Li, Wen-Jun

    2013-01-01

    A novel actinobacterial strain, MBRL 353(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. Comparison of 16S rRNA gene sequences of strain MBRL 353(T) and other members of the genus Rhodococcus showed sequence similarities ranging from 95.5 to 98.2 %, with strain MBRL 353(T) showing closest sequence similarity to Rhodococcus triatomae IMMIB RIV-085(T) (98.2 %) and Rhodococcus equi DSM 20307(T) (97.2 %). DNA-DNA hybridization results, however, revealed that DNA-DNA relatedness values between strain MBRL 353(T) and R. triatomae DSM 44892(T) (43.4 %) and R. equi DSM 20307(T) (33.4 %) were well below the 70 % limit for species identification. Strain MBRL 353(T) contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose and arabinose in the cell wall. Mycolic acids were present. The major fatty acids were C(16 : 0) (45.7 %), C(18 : 1)ω9c (18.2 %) and 10-methyl C(18 : 0) (11.3 %). The only menaquinone detected was MK-8(H(2)), while the major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and one unknown phospholipid. The G+C content of the genomic DNA was 69.2 mol%. The phenotypic and genotypic data showed that strain MBRL 353(T) merits recognition as a representative of a novel species of the genus Rhodococcus for which the name Rhodococcus canchipurensis sp. nov. is proposed; the type strain is MBRL 353(T) (= KCTC 19851(T) = JCM 17578(T)).

  3. Biotransformation of ginsenoside Rb1 to ginsenoside C-K by endophytic fungus Arthrinium sp. GE 17-18 isolated from Panax ginseng.

    PubMed

    Fu, Y; Yin, Z-H; Wu, L-P; Yin, C-R

    2016-09-01

    This research aimed to isolate β-glycosidase-producing endophytic fungus in Panax ginseng to achieve biotransformation of ginsenoside Rb1 to ginsenoside C-K. Of these 15 β-glucosidase-producing endophytic fungus isolated from ginseng roots, a β-glucosidase-producing endophytic fungi GE 17-18 could hydrolyse major ginsenosides Rb1 to minor ginsenoside C-K with metabolic pathways: ginsenoside Rb1→ginsenoside Rd→ginsenoside F2→ginsenoside C-K. Phylogenetic analysis of ITS gene sequences indicated that the strain GE 17-18 belongs to the genus Arthrinium and is most closely related to Arthrinium sp. HQ832803.1. This is the first study to provide information of cultivable β-glycosidase-producing Endophytic fungus in Panax ginseng. The strain GE 17-18 has potential to be applied on the preparation for minor ginsenoside C-K in pharmaceutical industry. © 2016 The Society for Applied Microbiology.

  4. Natural Products from Mangrove Actinomycetes

    PubMed Central

    Xu, Dong-Bo; Ye, Wan-Wan; Han, Ying; Deng, Zi-Xin; Hong, Kui

    2014-01-01

    Mangroves are woody plants located in tropical and subtropical intertidal coastal regions. The mangrove ecosystem is becoming a hot spot for natural product discovery and bioactivity survey. Diverse mangrove actinomycetes as promising and productive sources are worth being explored and uncovered. At the time of writing, we report 73 novel compounds and 49 known compounds isolated from mangrove actinomycetes including alkaloids, benzene derivatives, cyclopentenone derivatives, dilactones, macrolides, 2-pyranones and sesquiterpenes. Attractive structures such as salinosporamides, xiamycins and novel indolocarbazoles are highlighted. Many exciting compounds have been proven as potential new antibiotics, antitumor and antiviral agents, anti-fibrotic agents and antioxidants. Furthermore, some of their biosynthetic pathways have also been revealed. This review is an attempt to consolidate and summarize the past and the latest studies on mangrove actinomycetes natural product discovery and to draw attention to their immense potential as novel and bioactive compounds for marine drugs discovery. PMID:24798926

  5. Alkaloids and polyketides from Penicillium citrinum, an endophyte isolated from the Moroccan plant Ceratonia siliqua.

    PubMed

    El-Neketi, Mona; Ebrahim, Weaam; Lin, Wenhan; Gedara, Sahar; Badria, Farid; Saad, Hassan-Elrady A; Lai, Daowan; Proksch, Peter

    2013-06-28

    The endophytic fungus Penicillium citrinum was isolated from a fresh stem of the Moroccan plant Ceratonia siliqua. Extracts of P. citrinum grown on rice and white bean media yielded five new compounds, namely, citriquinochroman (1), tanzawaic acids G and H (2 and 3), 6-methylcurvulinic acid (4), 8-methoxy-3,5-dimethylisoquinolin-6-ol (5), and one new natural product, 1,2,3,11b-tetrahydroquinolactacide (6), which had previously been described as a synthetic compound. In addition, 13 known compounds including seven alkaloids and six polyketides were isolated. The structures of the new compounds were unambiguously determined on the basis of one- and two-dimensional NMR spectroscopy as well as by high-resolution mass spectrometry. Citriquinochroman (1) features a new skeleton, consisting of quinolactacide and (3S)-6-hydroxy-8-methoxy-3,5-dimethylisochroman linked by a C-C bond. 1,2,3,11b-Tetrahydroquinolactacide (6) may be a biogenetic precursor of quinolactacide. Citriquinochroman (1) showed cytotoxicity against the murine lymphoma L5178Y cell line with an IC50 value of 6.1 μM, while the other compounds were inactive (IC50 >10 μM) in this assay.

  6. Biogeography of symbiotic and other endophytic bacteria isolated from medicinal Glycyrrhiza species in China.

    PubMed

    Li, Li; Sinkko, Hanna; Montonen, Leone; Wei, Gehong; Lindström, Kristina; Räsänen, Leena A

    2012-01-01

    A total of 159 endophytic bacteria were isolated from surface-sterilized root nodules of wild perennial Glycyrrhiza legumes growing on 40 sites in central and northwestern China. Amplified fragment length polymorphism (AFLP) genomic fingerprinting and sequencing of partial 16S rRNA genes revealed that the collection mainly consisted of Mesorhizobium, Rhizobium, Sinorhizobium, Agrobacterium and Paenibacillus species. Based on symbiotic properties with the legume hosts Glycyrrhiza uralensis and Glycyrrhiza glabra, we divided the nodulating species into true and sporadic symbionts. Five distinct Mesorhizobium groups represented true symbionts of the host plants, the majority of strains inducing N2-fixing nodules. Sporadic symbionts consisted of either species with infrequent occurrence (Rhizobium galegae, Rhizobium leguminosarum) or species with weak (Sinorhizobium meliloti, Rhizobium gallicum) or no N2 fixation ability (Rhizobium giardinii, Rhizobium cellulosilyticum, Phyllobacterium sp.). Multivariate analyses revealed that the host plant species and geographic location explained only a small part (14.4%) of the total variation in bacterial AFLP patterns, with the host plant explaining slightly more (9.9%) than geography (6.9%). However, strains isolated from G. glabra were clearly separated from those from G. uralensis, and strains obtained from central China were well separated from those originating from Xinjiang in the northwest, indicating both host preference and regional endemism.

  7. Molecular profiling and bioactive potential of an endophytic fungus Aspergillus sulphureus isolated from Sida acuta: a medicinal plant.

    PubMed

    Murali, M; Mahendra, C; Hema, P; Rajashekar, N; Nataraju, A; Sudarshana, M S; Amruthesh, K N

    2017-12-01

    Sida acuta Burm.f. (Malvaceae) extracts are reported to have applications against malaria, diuretic, antipyretic, nervous and urinary diseases. No fungal endophytes of S. acuta are reported. Isolation, identification and evaluation of antibacterial, antioxidant, anticancer and haemolytic potential of fungal endophytes from the ethnomedcinal plant S. acuta. Sida acuta stem segments were placed on PDA medium to isolate endophytic fungi. The fungus was identified by genomic DNA analysis and phylogenetic tree was constructed using ITS sequences (GenBank) to confirm species. The antibacterial efficacy of Aspergillus sulphureus MME12 ethyl acetate extract was tested against Gram-positive and Gram-negative pathogenic bacteria. DPPH free radical scavenging activity, anticancer and DNA fragmentation against EAC cells, and direct haemolytic activity (100-500 μg/mL) using human erythrocytes were determined. The ethyl acetate extract of A. sulphureus (Fresen.) Wehmer (Trichocomaceae) demonstrated significant antibacterial potential against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Salmonella typhi compared to streptomycin. MIC against test pathogens was in the range of 15.6-62.5 μg/mL. The antioxidant results revealed significant RSA from 12.43% to 62.02% (IC50 = 350.4 μg/mL, p ≤ 0.05). MME12 offered considerable inhibition of EAC proliferation (23% to 84%, IC50 = 216.7 μg/mL, p ≤ 0.05) supported by DNA fragmentation studies. The extract also offered insignificant haemolysis (5.6%) compared to Triton X-100. A single endophytic fungus, A. sulphureus MME12 was isolated and identified using molecular profiling. The above-mentioned findings support the pharmacological application of A. sulphureus MME12 extract and demand for purification of the active principle(s).

  8. Endophytic Trichoderma isolates from tropical environments delay disease onset and induce resistance against Phytophthora capsici in hot pepper using multiple mechanisms

    USDA-ARS?s Scientific Manuscript database

    Isolates of several Trichoderma spp., were collected from tropical environments as potential biocontrol agents for cacao (Theobroma cacao) diseases. The diversity of isolates collected, including new species, and there endophytic nature on their host plants, led us to consider if these isolates have...

  9. Isolation, Colonization, and Chlorpyrifos Degradation Mediation of the Endophytic Bacterium Sphingomonas Strain HJY in Chinese Chives (Allium tuberosum).

    PubMed

    Feng, Fayun; Ge, Jing; Li, Yisong; Cheng, Jinjin; Zhong, Jianfeng; Yu, Xiangyang

    2017-02-15

    The endophyte-plant interaction can benefit the host in many different ways. An endophytic bacterium strain (HJY) capable of degrading chlorpyrifos (CP) was isolated from Chinese chives (Allium tuberosum Rottl. ex Spreng). The isolated bacterium HJY classified as Sphingomonas sp. strain HJY could use CP as the sole carbon source. After being marked with the gfp gene, the colonization and distribution of strain HJY-gfp were directly observed in different tissues of Chinese chives with a confocal laser scanning microscope. The inoculation of strain HJY-gfp in Chinese chives resulted in a higher degradation of CP inside the plants than in uninoculated plants. With drench application, up to 70 and 66% of CP were removed from shoots and roots of inoculated Chinese chives, respectively. Moreover, up to 75% of CP was removed from the soil containing plants inoculated with HJY-gfp. With foliage application, the applied concentration of chlorpyrifos affected the degradation performance of strain HJY in Chinese chives. Significant differences were observed only between inoculated and uninoculated Chinese chives with the low applied concentration of CP. Together, other than natural endophyte-assisted plant protection for food safety, the interaction of HJY and plant may be also a promising strategy for in situ bioremediation of soil contaminated with CP.

  10. Production of potent antimicrobial agent by actinomycete, Streptomyces sannanensis strain SU118 isolated from phoomdi in Loktak Lake of Manipur, India.

    PubMed

    Singh, Laishram Shantikumar; Sharma, Hemant; Talukdar, Narayan Chandra

    2014-11-19

    Actinomycetes have provided a wealth of bioactive secondary metabolites with interesting activities such as antimicrobial, antiviral and anticancer. The study aims at isolation, characterization and the antimicrobial potentiality of Streptomyces sannanensis SU118 obtained from Phoomdi, a unique habitat of Loktak Lake of Manipur, India. An actinomycete strain isolated from Phoomdi soil of Loktak Lake of Manipur, India was identified as Streptomyces sannanensis SU118. It is a Gram-positive filamentous bacterium which exhibits antimicrobial activity only against Gram-positive bacteria, while Gram-negative organisms were not affected. Glucose Soyabean meal broth was found to be the suitable medium for antibiotic production at 28°C for seven days of incubation. The antimicrobial agent produced by the strain was extracted with ethyl acetate as solvent and purified by thin layer chromatography. Screening and bioassay - guided fractionation of the ethyl acetate extract from the culture filtrate led to the isolation of an active potential compound (R f value 0.56) with λmax 275.0 nm which has got the lowest minimum inhibitory concentration (0.5 μg/ml) against Staphylococcus aureus MTCC 96 and Staphylococcus aureus (clinical isolate), whereas highest (3.0 μg/ml) was recorded against Mycobacterium smegmatis MTCC 6 and Bacillus circulans MTCC 8074. This study has therefore uncovered the potential of exploring virgin untapped habitats in the Indo-Burma biodiversity hot spot region as reservoir of promising antimicrobial metabolite producer. These results highlighted the scope for further characterization of the metabolite and could be a candidate in the generation of new antimicrobial agents.

  11. [Isolation, identification and anticancer activity of an endophytic fungi from Juglans mandshurica].

    PubMed

    Li, Meiya; Wu, Yunwei; Jiang, Fusheng; Yu, Xiangli; Tang, Kexuan; Miao, Zhiqi

    2009-07-01

    The endophytic fungus named FSN006 was isolated from the inner bark of Juglans mandshurica. It grew quickly and formed circular colony on PDA plate. The upper side of the colony was white, while the lower side of the colony and the conditioned medium were light yellow as a result of significant yellow pigment substances were produced and secreted by the fungi. Green elliptic conidia appeared when cultured on CMX plate. Based on the morphology identification and ITS sequence, it was clear that this fungus belonged to the Deuteromycotina, HyPhomycetes, Moniliales, Trichoderma longibrachiatum. The conditioned medium of FSN006 showed a high anti-tumor ability against liver cancer cell-HepG2, and reached its IC50 concentration after being diluted 20 times, while the IC50 concentration of curcumine was(11.49 +/- 0.12) mg x L(-1). In addition, there was preeminent selective inhibiting effect against the normal liver cell strain HL-7702 and its caner counter strain HepG2. The inhibiting effect against strain HL-7702 was only one quarter of that against HepG2 at the concentration of IC50. Therefore, the fermentation of FSN006 may provide a possible way to produce anticancer drug with higher efficiency and lower toxicity.

  12. Prauserella endophytica sp. nov., an endophytic actinobacterium isolated from Tamarix taklamakanensis.

    PubMed

    Liu, Jia-Meng; Habden, Xugela; Guo, Lin; Tuo, Li; Jiang, Zhong-Ke; Liu, Shao-Wei; Liu, Xian-Fu; Chen, Li; Li, Rong-Feng; Zhang, Yu-Qin; Sun, Cheng-Hang

    2015-06-01

    A novel endophytic actinobacterium, designated strain SP28S-3(T), was isolated from a surface-sterilized stem of Tamarix taklamakanensis collected from the southern edge of Taklamakan desert, Xinjiang, China. Strain SP28S-3(T) was found to show chemotaxonomic and morphological properties consistent with its classification in the genus Prauserella. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphoglycolipid, phosphatidylcholine, phosphatidylinositol, a glycolipid, an aminolipid and unidentified phospholipids. The major fatty acids (>10 %) were identified as iso-C16:0 and C16:0. The genomic DNA G+C content was determined to be 69.7 mol%. Phylogenetic analysis of strain SP28S-3(T) clearly showed that the strain had the highest similarity of 16S rRNA gene sequence with Prauserella coralliicola SCSIO 11529(T) (99.9 %), followed by Prauserella marina DSM 45268(T) (97.0 %) and is affiliated with the genus Prauserella. The low level (47.8 ± 5.5 %) of DNA-DNA relatedness between strain SP28S-3(T) and P. coralliicola SCSIO 11529(T) combined with other polyphasic taxonomic evidence clearly support the conclusion that strain SP28S-3(T) represents a novel Prauserella species, for which the name Prauserella endophytica sp. nov. is proposed. The type strain is SP28S-3(T) (=DSM 46655(T) = CGMCC 4.7182 (T)).

  13. Trichoderma gamsii (NFCCI 2177): a newly isolated endophytic, psychrotolerant, plant growth promoting, and antagonistic fungal strain.

    PubMed

    Rinu, K; Sati, Priyanka; Pandey, Anita

    2014-05-01

    An endophytic fungus has been isolated from the lateral roots of lentil (Lens esculenta Moench), growing under mountain ecosystem of Indian Himalayan Region (IHR). While the fungus was observed as fast growing with white scanty mycelium turning to turmeric brown in 5 days of incubation at 25 °C, it also produced a unique odor. The fungus exhibited growth between 4 and 30 °C (optimum 25 °C) and tolerated pH between 2.0 and 13.5 (optimum 4-6). Based on phenotypic (colony morphology and microscopy) and genotypic (18S rRNA analysis) characters, the fungus was identified as Trichoderma gamsii (99% similarity). The fungus was evaluated for its plant growth promotion and biocontrol properties. The fungus was found to be positive for phosphate solubilization, chitinase activity, and production of ammonia and salicylic acid, while the results for production of IAA, HCN, and siderophores were negative. Out of the seven phytopathogenic fungi tested, it showed antagonism against six. Bioassays conducted under green house using four test crops (two cereals and two legumes) showed its potential in plant growth promotion. The fungus has potential to be developed as a bioformulation for application under mountain ecosystem.

  14. Bioactive metabolites from the endophytic fungus Stemphylium globuliferum isolated from Mentha pulegium.

    PubMed

    Debbab, Abdessamad; Aly, Amal H; Edrada-Ebel, Ruangelie; Wray, Victor; Müller, Werner E G; Totzke, Frank; Zirrgiebel, Ute; Schächtele, Christoph; Kubbutat, Michael H G; Lin, Wen Han; Mosaddak, Mahjouba; Hakiki, Abdelhak; Proksch, Peter; Ebel, Rainer

    2009-04-01

    The endophytic fungus Stemphylium globuliferum was isolated from stem tissues of the Moroccan medicinal plant Mentha pulegium. Extracts of the fungus, which was grown on solid rice medium, exhibited considerable cytotoxicity when tested in vitro against L5178Y cells. Chemical investigation yielded five new secondary metabolites, alterporriol G (4) and its atropisomer alterporriol H (5), altersolanol K (11), altersolanol L (12), stemphypyrone (13), and the known compounds 6-O-methylalaternin (1), macrosporin (2), altersolanol A (3), alterporriol E (6), alterporriol D (7), alterporriol A (8), alterporriol B (9), and altersolanol J (10). The structures were determined on the basis of one- and two-dimensional NMR spectroscopy and mass spectrometry. Among the alterporriol-type anthranoid dimers, the mixture of alterporriols G and H (4/5) exhibited considerable cytotoxicity against L5178Y cells with an EC(50) value of 2.7 microg/mL, whereas the other congeners showed only modest activity. The compounds were also tested for kinase inhibitory activity in an assay involving 24 different kinases. Compounds 1, 2, 3, and the mixture of 4 and 5 were the most potent inhibitors, displaying EC(50) values between 0.64 and 1.4 microg/mL toward individual kinases.

  15. Isolation and biological activities of an endophytic Mortierella alpina strain from the Antarctic moss Schistidium antarctici.

    PubMed

    Melo, Itamar S; Santos, Suikinai N; Rosa, Luiz H; Parma, Marcia M; Silva, Leonardo J; Queiroz, Sonia C N; Pellizari, Vivian H

    2014-01-01

    The Antarctic endophytic fungus (strain ITA1-CCMA 952) was isolated from the moss Schistidium antarctici found in Admiralty Bay, King George Island, Antarctica. Strain ITA1-CCMA 952 was assigned to the specie Mortierella alpina by phylogenetic analysis based on 18S rRNA gene sequences. This strain produces high levels of polyunsaturated fatty acids (PUFAs), including y-(gamma) linolenic acid and arachidonic acid, which when combined represents 48.3% of the total fatty acid content. Fungal extracts demonstrated strong antioxidant activity with the EC50 value of 48.7 μg mL(-1) and also a strong antibacterial activity, mainly against the following bacteria: Escherichia coli, with a MIC of 26.9 μg mL(-1) and Pseudomonas aeruginosa and Enterococcus faecalis, both with a MIC of 107 μg mL(-1). A GC-MS analysis of the chloroform fraction obtained from the crude extract revealed the presence of potential antimicrobials (Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl) and Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(phenylmethyl)) as the major compounds. Therefore, the M. alpina strain ITA1-CCMA 952 is a promising fungus for the biotechnological production of antibiotics, antioxidant substances and PUFAs. This study highlights the need for more research in extreme environments, such as Antarctica.

  16. [Isolation of two endophytic phenanthrene-degrading strains and their degradation capacity].

    PubMed

    Ni, Xue; Liu, Juan; Gao, Yan-Zheng; Zhu, Xue-Zhu; Sun, Kai

    2013-02-01

    Two endophytic bacterial strains, which could degrade high concentration (up to 200 mg.L-1) of phenanthrene in liquid, were isolated from plants grown in PAHs-contaminated soils by the selective. enrichment culture. According to the results of morphology, physiology and the phylogenetic analyses of 16S rDNA sequence, stain P1 was identified as Stenotrophomonas sp. , and strain P3 was identified as Pseudomonas sp.. Two strains were aerobic bacteria, the degradation rates of phenanthrene (100 mg.L-1) by strain P1 and strain P3 were all greater than 90% at 28 degrees C on the rotation shaker at 150 r.min-1 for 7 days. The degradation rates of phenanthrene by two strains were greater than 70% when cultivated under the conditions as: 20-30 degrees C , pH 6-8, 0%-4% NaCl, 10-30 mL/100 mL inventory. It suggested that the optimum culture condition was: 30 degrees C, pH 7.0, NaCl< or =4% , inventory < or = 30 mL/100 mL flask. Through comprehensive comparison analyses on the degradation capacity of two strains, it showed that the tolerance of strain P1 to high temperature was higher than that of str ain P3, while the tolerance of strain P3 to pH change and anoxic condition was higher than that of strain P1.

  17. Mycoparasitism of endophytic fungi isolated from reed on soilborne phytopathogenic fungi and production of cell wall-degrading enzymes in vitro.

    PubMed

    Cao, Ronghua; Liu, Xiaoguang; Gao, Kexiang; Mendgen, Kurt; Kang, Zhensheng; Gao, Jianfeng; Dai, Yang; Wang, Xue

    2009-12-01

    Antagonism of three endophytic fungi isolated from common reed (Phragmites australis) against eight soilborne pathogenic fungi was investigated on potato dextrose agar by light microscopy, scanning electron microscopy, and transmission electron microscopy. Inhibitory zones were not observed. The microscopical studies suggested that the endophytes inhibit growth of soilborne pathogens by means of coiling around hyphae and, after penetration, the degradation of hyphal cytoplasm. Since penetration of hyphae seems to play a major role in parasitism, we studied the production of cell wall degrading enzymes by the three endophytes. Choiromyces aboriginum produced higher activities of beta-1,3-glucanases compared to Stachybotrys elegans and Cylindrocarpon sp. For C. aboriginum and S. elegans, colloidal chitin was the best substrate for the induction of beta-1,3-glucanases and chitinases, respectively. This result suggests that mycoparasitism by endophytes on soilborne plant pathogens can be explained by their mycoparasitic activity.

  18. Renaissance in antibacterial discovery from actinomycetes.

    PubMed

    Baltz, Richard H

    2008-10-01

    The soil actinomycetes have been important sources of antibiotics, but were nearly abandoned in recent years in favor of high-throughput target-based screening of chemical libraries. The latter approach has not been productive, so it is time to reinvigorate the discovery of new antibiotics from a proven source. Recent progress has been made on antibiotic discovery from actinomycetes by using high-throughput fermentation, isolation of marine actinomycetes, mining genomes for cryptic pathways, and combinatorial biosynthesis to generate new secondary metabolites related to existing pharmacophores.

  19. Endophytic fungi associated with cacti in Arizona.

    PubMed

    Suryanarayanan, Trichur S; Wittlinger, Sally K; Faeth, Stanley H

    2005-05-01

    21 cactus species occurring in various localities within Arizona were screened for the presence of fungal endophytes. 900 endophyte isolates belonging to 22 fungal species were isolated. Cylindropuntia fulgida had the maximum endophyte species diversity, while C. ramosissima harboured the maximum number of endophyte isolates. Alternaria sp., Aureobasidium pullulans, and Phoma spp. were isolated from several cactus species. The diversity of the endophyte assemblages was low and no host specificity among endophytes was observed. However, the frequencies of colonization of the few endophyte species recovered were high and comparable to those reported for tropical plant hosts. Species of Colletotrichum, Phomopsis, and Phyllosticta, which are commonly isolated as endophytes from plants of more mesic habitats, were absent from these cacti.

  20. Phytohormone production endowed with antagonistic potential and plant growth promoting abilities of culturable endophytic bacteria isolated from Clerodendrum colebrookianum Walp.

    PubMed

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Leo, Vincent Vineeth; Gupta, Vijai Kumar; Singh, Bhim Pratap

    2016-12-01

    In this study, culturable endophytic bacterial isolates obtained from an ethnomedicinal plant Clerodendrum colebrookianum Walp., were assessed for their diversity, in vitro screening for their plant growth promoting (PGP) activities and to use them as inoculant for in vivo PGP activities with biocontrol potential. Totally, 73 isolates were recovered from different tissues of C. colebrookianum were identified by 16S rRNA gene sequencing and phylogenetically analyzed by using BOX-PCR fingerprinting. Out of 73 isolates, 52 exhibited varying extents of antagonistic potential were selected for screening for various PGP traits. Concerning the PGP activities, the percentage of isolates positive for P-solubilisation, indolic compounds production, siderophore and ammonia production were 84.6, 92.3, 78.8 and 98.0 respectively. All isolates were positive for the production of hydrocyanic acid (HCN) and 86.5%, 84.6% and 90.3% of isolates showed significant cellulase, amylase and protease production respectively. Further, the top 10 bacterial isolates based on a bonitur scale with multiple PGP activities were screened for root surface colonization and biofilm formation ability. Out of selected 10 isolates, 9 showed significant potential for root surface colonization on tomato roots. Isolate BPSAC6 identified as Bacillus sp. was most efficient in biofilm formation as assessed with respect to the intensity of crystal violet, which further showed their potential to withstand various biotic and abiotic stresses. Furthermore, Bacillus sp. strain BPSAC6 showed a significant increase in shoot and root height as well as fresh weight after 45 and 60 d of inoculation with tomato seedlings. Additionally, biosynthetic potential of antagonistic isolate was detection by using PKSI, PKSII and NRPS biosynthetic genes. Two isolates Pseudomonas psychrotolerans and Labrys wisconsinensis were reported first time as an endophyte. At last, first time an endophytic bacterial strain Bacillus sp. BPSAC

  1. Detection and identification of novel actinomycetes.

    PubMed

    Williams, S T; Locci, R; Beswick, A; Kurtböke, D I; Kuznetsov, V D; Le Monnier, F J; Long, P F; Maycroft, K A; Palma, R A; Petrolini, B

    1993-10-01

    The actinomycetes are well known as a group of filamentous, Gram-positive bacteria that produce many useful secondary metabolites, including antibiotics and enzymes. Although they have been intensively studied for both theoretical and practical objectives, there is much scope for developing our basic knowledge of the means of detection and isolation of these microbes. This session concentrated on new methods for the detection and identification of novel actinomycetes from a range of environments. Approaches to the detection of actinomycetes ranged from investigations of neglected habitats and extreme environments (e.g. alkaline soils and oil drills) to the analysis of DNA extracted from the environment and use of specific phages. The continuing problems of the identification of actinomycete isolates were also considered. Topics discussed included use of phage typing, DNA probes, and correlation between phenetic and genotypic species of Streptomyces.

  2. Endophytic Fungi from Paddy

    PubMed Central

    Zakaria, Latiffah; Yaakop, Amira Suriaty; Salleh, Baharuddin; Zakaria, Maziah

    2010-01-01

    Endophytic fungi were isolated from different parts of healthy paddy plants (Oryza sativa). The most common endophytic fungal genus recovered was Fusarium, followed by Aspergillus, Curvularia, Penicillium, Gilmaniella and Arthrobotrys foliicola. Fusarium and Curvularia had higher occurrences in the seeds compared with the other fungi. Aspergillus was recovered mostly from leaf blades and Penicillium from the leaf sheath. Gilmaniella and A. foliicola were isolated only from the roots and leaf blade, respectively. The assemblage of endophytic fungi in healthy tissues of paddy plants may indicate that some of the fungi are possible latent pathogens and some may become saprophytic. PMID:24575194

  3. New method for RNA isolation from actinomycetes: application to the transcriptional analysis of the alcohol oxidoreductase gene thcE in Rhodococcus and Mycobacterium.

    PubMed

    Nagy, I; Schoofs, G; De Schrijver, A; Vanderleyden, J; De Mot, R

    1997-07-01

    A new protocol for the isolation of RNA from Rhodococcus and other actinomycetes such as Mycobacterium and Amycolatopsis was developed. The method is based on rapid lysis of cells in a high-speed reciprocal shaker using small abrasive particles followed by spin column purification of the lysate. This quick procedure yields RNA preparations suitable for functional studies. This was shown for the thcE gene of R. erythropolis NI86/21, which encodes a N,N'-dimethyl-4-nitrosoaniline-dependent alcohol oxidoreductase. The thcE transcript was detected by Northern hybridization in R. erythropolis, R. fascians, Mycobacterium chlorophenolicum and Mycobacterium smegmatis. The transcriptional start point of the gene was determined by primer extension of the R. erythropolis mRNA.

  4. Improved plant growth and Zn accumulation in grains of rice (Oryza sativa L.) by inoculation of endophytic microbes isolated from a Zn Hyperaccumulator, Sedum alfredii H.

    PubMed

    Wang, Yuyan; Yang, Xiaoe; Zhang, Xincheng; Dong, Lanxue; Zhang, Jie; Wei, Yanyan; Feng, Ying; Lu, Lingli

    2014-02-26

    This study is to investigate the possibility of zinc (Zn) biofortification in the grains of rice (Oryza sativa L.) by inoculation of endophytic strains isolated from a Zn hyperaccumulator, Sedum alfredii Hance. Five endophytic strains, Burkholderia sp. SaZR4, Burkholderia sp. SaMR10, Sphingomonas sp. SaMR12, Variovorax sp. SaNR1, and Enterobacter sp. SaCS20, isolated from S. alfredii, were inoculated in the roots of Japonica rice Nipponbare under hydroponic condition. Fluorescence images showed that endophytic strains successfully colonized rice roots after 72 h. Improved root morphology and plant growth of rice was observed after inoculation with endophytic strains especially SaMR12 and SaCS20. Under hydroponic conditions, endophytic inoculation with SaMR12 and SaCS20 increased Zn concentration by 44.4% and 51.1% in shoots, and by 73.6% and 83.4% in roots, respectively. Under soil conditions, endophytic inoculation with SaMR12 and SaCS20 resulted in an increase of grain yields and elevated Zn concentrations by 20.3% and 21.9% in brown rice and by 13.7% and 11.2% in polished rice, respectively. After inoculation of SaMR12 and SaCS20, rhizosphere soils of rice plants contained higher concentration of DTPA-Zn by 10.4% and 20.6%, respectively. In situ micro-X-ray fluorescence mapping of Zn confirmed the elevated Zn content in the rhizosphere zone of rice treated with SaMR12 as compared with the control. The above results suggested that endophytic microbes isolated from S. alfredii could successfully colonize rice roots, resulting in improved root morphology and plant growth, increased Zn bioavailability in rhizosphere soils, and elevated grain yields and Zn densities in grains.

  5. An Endophytic Fungus, Talaromyces radicus, Isolated from Catharanthus roseus, Produces Vincristine and Vinblastine, Which Induce Apoptotic Cell Death.

    PubMed

    Palem, Padmini P C; Kuriakose, Gini C; Jayabaskaran, Chelliah

    2015-01-01

    Endophytic fungi isolated from Catharanthus roseus were screened for the production of vincristine and vinblastine. Twenty-two endophytic fungi isolated from various tissues of C. roseus were characterized taxonomically by sequence analysis of the internal transcribed spacer (ITS) region of rDNA and grouped into 10 genera: Alternaria, Aspergillus, Chaetomium, Colletotrichum, Dothideomycetes, Eutypella, Eutypa, Flavodon, Fusarium and Talaromyces. The antiproliferative activity of these fungi was assayed in HeLa cells using the MTT assay. The fungal isolates Eutypella sp--CrP14, obtained from stem tissues, and Talaromyces radicus--CrP20, obtained from leaf tissues, showed the strongest antiproliferative activity, with IC50 values of 13.5 μg/ml and 20 μg/ml, respectively. All 22 endophytic fungi were screened for the presence of the gene encoding tryptophan decarboxylase (TDC), the key enzyme in the terpenoid indole alkaloid biosynthetic pathway, though this gene could only be amplified from T. radicus--CrP20 (NCBI GenBank accession number KC920846). The production of vincristine and vinblastine by T. radicus--CrP20 was confirmed and optimized in nine different liquid media. Good yields of vincristine (670 μg/l) in modified M2 medium and of vinblastine (70 μg/l) in potato dextrose broth medium were obtained. The cytotoxic activity of partially purified fungal vincristine was evaluated in different human cancer cell lines, with HeLa cells showing maximum susceptibility. The apoptosis-inducing activity of vincristine derived from this fungus was established through cell cycle analysis, loss of mitochondrial membrane potential and DNA fragmentation patterns.

  6. An Endophytic Fungus, Talaromyces radicus, Isolated from Catharanthus roseus, Produces Vincristine and Vinblastine, Which Induce Apoptotic Cell Death

    PubMed Central

    Jayabaskaran, Chelliah

    2015-01-01

    Endophytic fungi isolated from Catharanthus roseus were screened for the production of vincristine and vinblastine. Twenty-two endophytic fungi isolated from various tissues of C. roseus were characterized taxonomically by sequence analysis of the internal transcribed spacer (ITS) region of rDNA and grouped into 10 genera: Alternaria, Aspergillus, Chaetomium, Colletotrichum, Dothideomycetes, Eutypella, Eutypa, Flavodon, Fusarium and Talaromyces. The antiproliferative activity of these fungi was assayed in HeLa cells using the MTT assay. The fungal isolates Eutypella sp—CrP14, obtained from stem tissues, and Talaromyces radicus—CrP20, obtained from leaf tissues, showed the strongest antiproliferative activity, with IC50 values of 13.5 μg/ml and 20 μg/ml, respectively. All 22 endophytic fungi were screened for the presence of the gene encoding tryptophan decarboxylase (TDC), the key enzyme in the terpenoid indole alkaloid biosynthetic pathway, though this gene could only be amplified from T. radicus—CrP20 (NCBI GenBank accession number KC920846). The production of vincristine and vinblastine by T. radicus—CrP20 was confirmed and optimized in nine different liquid media. Good yields of vincristine (670 μg/l) in modified M2 medium and of vinblastine (70 μg/l) in potato dextrose broth medium were obtained. The cytotoxic activity of partially purified fungal vincristine was evaluated in different human cancer cell lines, with HeLa cells showing maximum susceptibility. The apoptosis-inducing activity of vincristine derived from this fungus was established through cell cycle analysis, loss of mitochondrial membrane potential and DNA fragmentation patterns. PMID:26697875

  7. Diversity of sporadic symbionts and nonsymbiotic endophytic bacteria isolated from nodules of woody, shrub, and food legumes in Ethiopia.

    PubMed

    Aserse, Aregu Amsalu; Räsänen, Leena A; Aseffa, Fassil; Hailemariam, Asfaw; Lindström, Kristina

    2013-12-01

    Fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in Ethiopia: Crotalaria spp., Indigofera spp., and Erythrina brucei, and the food legumes soybean and common bean. Based on partial 16S rRNA gene sequence analysis, the majority of the isolates were identified as Gram-negative bacteria belonging to the genera Achromobacter, Agrobacterium, Burkholderia, Cronobacter, Enterobacter, Mesorhizobium, Novosphingobium, Pantoea, Pseudomonas, Rahnella, Rhizobium, Serratia, and Variovorax. Seven isolates were Gram-positive bacteria belonging to the genera Bacillus, Paenibacillus, Planomicrobium, and Rhodococcus. Amplified fragment length polymorphism (AFLP) fingerprinting showed that each strain was genetically distinct. According to phylogenetic analysis of recA, glnII, rpoB, and 16S rRNA gene sequences, Rhizobium, Mesorhizobium, and Agrobacterium were further classified into six different genospecies: Agrobacterium spp., Agrobacterium radiobacter, Rhizobium sp., Rhizobium phaseoli, Mesorhizobium sp., and putative new Rhizobium species. The strains from R. phaseoli, Rhizobium sp. IAR30, and Mesorhizobium sp. ERR6 induced nodules on their host plants. The other strains did not form nodules on their original host. Nine endophytic bacterial strains representing seven genera, Agrobacterium, Burkholderia, Paenibacillus, Pantoea, Pseudomonas, Rhizobium, and Serratia, were found to colonize nodules of Crotalaria incana and common bean on co-inoculation with symbiotic rhizobia. Four endophytic Rhizobium and two Agrobacterium strains had identical nifH gene sequences with symbiotic Rhizobium strains, suggesting horizontal gene transfer. Most symbiotic and nonsymbiotic endophytic bacteria showed plant growth-promoting properties in vitro, which indicate their potential role in the promotion of plant growth when colonizing plant roots and the rhizosphere.

  8. Endophytic Trichoderma isolates from tropical environments delay disease onset and induce resistance against Phytophthora capsici in hot pepper using multiple mechanisms.

    PubMed

    Bae, Hanhong; Roberts, Daniel P; Lim, Hyoun-Sub; Strem, Mary D; Park, Soo-Chul; Ryu, Choong-Min; Melnick, Rachel L; Bailey, Bryan A

    2011-03-01

    Endophytic Trichoderma isolates collected in tropical environments were evaluated for biocontrol activity against Phytophthora capsici in hot pepper (Capsicum annuum). Six isolates were tested for parasitic and antimicrobial activity against P. capsici and for endophytic and induced resistance capabilities in pepper. Isolates DIS 70a, DIS 219b, and DIS 376f were P. capsici parasites, while DIS 70a, DIS 259j, DIS 320c, and DIS 376f metabolites inhibited P. capsici. All six isolates colonized roots but were inefficient stem colonizers. DIS 259j, DIS 320c, and DIS 376f induced defense-related expressed sequence tags (EST) in 32-day-old peppers. DIS 70a, DIS 259j, and DIS 376f delayed disease development. Initial colonization of roots by DIS 259j or DIS 376f induced EST with potential to impact Trichoderma endophytic colonization and disease development, including multiple lipid transferase protein (LTP)-like family members. The timing and intensity of induction varied between isolates. Expression of CaLTP-N, encoding a LTP-like protein in pepper, in N. benthamiana leaves reduced disease development in response to P. nicotianae inoculation, suggesting LTP are functional components of resistance induced by Trichoderma species. Trichoderma isolates were endophytic on pepper roots in which, depending on the isolate, they delayed disease development by P. capsici and induced strong and divergent defense reactions.

  9. Bacillus capparidis sp. nov., an endophytic bacterium isolated from roots of Capparis spinosa L.

    PubMed

    Wang, Hong-Fei; Li, Qiu-Li; Zhang, Yong-Guang; Xiao, Min; Zhou, Xing-Kui; Guo, Jian-Wei; Duan, Yan-Qing; Li, Wen-Jun

    2017-02-01

    A novel endophytic bacterium, designated strain EGI 6500252T, was isolated from the surface-sterilized roots of a medicinal plant (Capparis spinosa L.) collected from Urumqi city, Xinjiang, north-west China. Cells were Gram-stain-positive, non-motile, aerobic, catalase- and oxidase-positive, rod-shaped and did not display spore formation. Strain EGI 6500252T grew at 10-40 °C (optimum 25-30 °C), at pH 6.0-8.0 (optimum pH 7.0) and in the presence of 0-10 % (w/v) NaCl (optimum 0-3 %). The major cellular fatty acids (>10 %) were identified as iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and summed feature 4. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, four unknown phospholipids, one unknown glycolipid and one unknown lipid. The dominant isoprenoid quinone was menaquinone 7 (MK-7). The DNA G+C content was 39.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain EGI 6500252T belonged to the genus Bacillus, and exhibited a highest 16S rRNA gene sequence similarity (96.2 %) that was lower than the suggested threshold (97.0 %) for separating bacterial species. On the basis of the phylogenetic analysis, chemotaxonomic data and physiological characteristics, strain EGI 6500252T represents a novel species of the genus Bacillus, for which the name Bacillus capparidis sp. nov. is proposed. The type strain is EGI 6500252T (=CGMCC 1.12820T=KCTC 33514T).

  10. Flavobacterium suaedae sp. nov., an endophyte isolated from the root of Suaeda corniculata.

    PubMed

    Sun, Ji-Quan; Xu, Lian; Liu, Min; Wang, Xin-Ying; Wu, Xiao-Lei

    2016-05-01

    A Gram-stain-negative, non-motile, yellow, endophytic bacterium, designated G16-7T, was isolated from the root of Suaeda corniculata in Inner Mongolia, northern China. Phylogenetic analysis, based on the 16S rRNA gene, revealed that strain G16-7T belonged to the genus Flavobacterium, with highest sequence similarities to Flavobacterium rakeshii FCS-5T, Flavobacterium suzhouense XIN-1T, Flavobacterium beibuense F44-8T, Flavobacterium hauense BX12T and Flavobacterium shanxiense YF-2, ranging from 92.7 % to 94.9 %. The predominant fatty acids of strain G16-7T were iso-C15 : 0, summed feature 3 (consisting of C16 : 1ω7c and/or C16 : 1ω6c), iso-C17 : 0 3-OH and summed feature 9 (consisting of iso-C17 : 1ω9c and/or C16 : 0 10-methyl), while MK-6 was the major respiratory quinone. The major polar lipids were phosphatidylethanolamine, an unknown phospholipid, an unknown aminophospholipid, four unknown aminolipids and three unknown lipids. The genomic DNA G+C content of the strain was 34.2 mol%. Based on the phenotypic and genotypic characteristics, strain G16-7T is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium suaedae sp. nov. is proposed. The type strain is G16-7T ( = CGMCC 1.15461T = KCTC 42947T).

  11. Rhizobium smilacinae sp. nov., an endophytic bacterium isolated from the leaf of Smilacina japonica.

    PubMed

    Zhang, Lei; Shi, Xu; Si, Meiru; Li, Changfu; Zhu, Lingfang; Zhao, Liang; Shen, Xihui; Wang, Yao

    2014-10-01

    During a study of endophytic bacteria from traditional Chinese medicinal plants, a bacterial strain, designated PTYR-5(T), was isolated from the leaf of Smilacina japonica A. Gray collected from Taibai Mountain in Shaanxi Province, north-west China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain PTYR-5(T) is a member of the genus Rhizobium, exhibiting the highest sequence similarities to R. cellulosilyticum LMG 23642(T) (97.2%), R. huautlense LMG 18254(T) (97.2%) and R. alkalisoli CCBAU 01393(T) (97.1%). The levels of 16S rRNA gene sequence similarity with respect to other Rhizobium species with validly published names were less than 97.0%. Phylogenies of the housekeeping genes atpD, recA and glnII confirmed its distinct position, showing low similarity with respect to those of recognized Rhizobium species (no more than 94.1, 90.0 and 88.0% similarity, respectively). The DNA-DNA relatedness values of strain PTYR-5(T) with R. cellulosilyticum LMG 23642(T), R. huautlense LMG 18254(T) and R. alkalisoli CCBAU 01393(T) were 33.6, 21.4 and 29.5 %, respectively. Based on phenotypic, phylogenetic and genotypic data, strain PTYR-5(T) is considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium smilacinae sp. nov. is proposed. The type strain is PTYR-5(T) (=CCTCC AB 2013016(T)=KCTC 32300(T)=LMG 27604(T)).

  12. Description of Micrococcus aloeverae sp. nov., an endophytic actinobacterium isolated from Aloe vera.

    PubMed

    Prakash, Om; Nimonkar, Yogesh; Munot, Hitendra; Sharma, Avinash; Vemuluri, Venkata Ramana; Chavadar, Mahesh S; Shouche, Yogesh S

    2014-10-01

    A yellow Gram-stain-positive, non-motile, non-endospore -forming, spherical endophytic actinobacterium, designated strain AE-6(T), was isolated from the inner fleshy leaf tissues of Aloe barbadensis (Aloe vera) collected from Pune, Maharashtra, India. Strain AE-6(T) grew at high salt concentrations [10% (w/v) NaCl], temperatures of 15-41 °C and a pH range of 5-12. It showed highest (99.7%) 16S rRNA gene sequence similarity with Micrococcus yunnanensis YIM 65004(T) followed by Micrococcus luteus NCTC 2665(T) (99.6%) and Micrococcus endophyticus YIM 56238(T) (99.0%). Ribosomal protein profiling by MALDI-TOF/MS also showed it was most closely related to M. yunnanensis YIM 65004(T) and M. luteus NCTC 2665(T). Like other members of the genus Micrococcus, strain AE-6(T) had a high content of branched chain fatty acids (iso-C15:0 and anteiso-C15:0). MK-8(H2) and MK-8 were the predominant isoprenoid quinones. Cell wall analysis showed an 'A2 L-Lys-peptide subunit' type of peptidoglycan and ribose to be the major cell wall sugar. The DNA G+C content was 70 mol%. Results of DNA-DNA hybridization of AE-6(T) with its closest relatives from the genus Micrococcus produced a value of less than 70%. Based on the results of this study, strain AE-6(T) could be clearly differentiated from other members of the genus Micrococcus. We propose that it represents a novel species of the genus Micrococcus and suggest the name Micrococcus aloeverae sp. nov., with strain AE-6(T) ( = MCC 2184(T) = DSM 27472(T)) as the type strain of the species. © 2014 IUMS.

  13. Streptomyces capparidis sp. nov., a novel endophytic actinobacterium isolated from fruits of Capparis spinosa L.

    PubMed

    Wang, Hong-Fei; Li, Qiu-Li; Xiao, Min; Zhang, Yong-Guang; Zhou, Xing-Kui; Narsing Rao, Manik Prabhu; Duan, Yan-Qing; Li, Wen-Jun

    2017-01-01

    A novel endophytic actinobacterial strain, designated EGI 6500195T, was isolated from fruits of Capparis spinosa. Growth occurred at 10-45 °C (optimum 30 °C), at pH 6-8 (optimum pH 7) and in the presence of 0-1 % (w/v) NaCl. Strain EGI 6500195T shared highest 16S rRNA gene sequence similarity (97.74 %) with Streptomyces vitaminophilus DSM 41686T and less than 97 % sequence similarity with other members of the genus Streptomyces. The diagnostic amino acid in the peptidoglycan was ll-diaminopimelic acid. Whole-cell hydrolysates contained glucose, ribose, fructose and mannose. The predominant menaquinones were MK-9(H6) and MK-9(H8). The polar lipid profile of strain EGI 6500195T included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylcholine, three unknown phospholipids, an unknown aminophospholipid and an unknown aminolipid. The cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0, iso-C16 : 0, anteiso-C17 : 1ω9c, summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B) and iso-C17 : 1ω9c. The DNA G+C content of strain EGI 6500195T was 74.1 mol%. The level of DNA-DNA relatedness between strain EGI 6500195T and Streptomyces. vitaminophilus DSM 41686T was 14.1±3.5 %. On the basis of the phenotypic, phylogenetic, chemotaxonomic and DNA-DNA hybridization data, strain EGI 6500195T represents a novel species of the genus Streptomyces, for which the name Streptomyces capparidis sp. nov. is proposed. The type strain is EGI 6500195T (=DSM 42145T=JCM 30089T).

  14. [Isolation and biodiversity of heavy metal tolerant endophytic bacteria from halotolerant plant species located in coastal shoal of Nantong].

    PubMed

    Bian, Guangkai; Zhang, Yueji; Qin, Sheng; Xing, Ke; Xie, Huansong; Jiang, Jihong

    2011-11-04

    We isolated and identified endophytic bacteria from halotolerant plants collected from coastal shoal of Nantong and investigated their heavy-metal tolerance and plant growth promoting potential. In total 45 strains were obtained from 4 halotolerant plants and 23 representative isolates were selected to detect their tolerance against NaCl and heavy metals of Cu2+, Pb2+, Cd2+, Zn2+, Hg2+; plant growth promoting index of nitrogen fixation, phosphate solubilization, indoleacetic acid (IAA) production and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase. Most of the isolates could grow under high consistency of Cu2+ and Pb2+. Of the bacteria 26.1% had the ability of nitrogen fixation, 21.7% of phosphate solubilization, 60.9% of IAA production and 39.1% of ACC deaminase activity. The results of 16S rRNA sequencing show that they belonged to the genera of Bacillus, Halobacillus, Oceanobacillus, Exiguobacterium, Serratia, Brevundimonas, Vibrio and Staphylococcus. Among them, strains KLBMP 2432 and KLBMP 2447 were potential novel species. The halotolerant plants located in the area of coastal shoal contain a variety of endophytic bacteria as well as the source of novel taxa. Some of them had the ability of plant growth promoting and high resistance against heavy-metal Cu2+ and Pb2+.

  15. Anticancer activity test of ethyl acetate extract of endophytic fungi isolated from soursop leaf (Annona muricata L.).

    PubMed

    Minarni; Artika, I Made; Julistiono, Heddy; Bermawie, Nurliani; Riyanti, Eny Ida; Hasim; Hasan, Akhmad Endang Zainal

    2017-06-01

    To analyze anticancer activity of an ethyl acetate extract of endophytic fungi isolated from soursop leaf (Annona muricata L.). Anticancer activity of fungal extracts was determined by observing its toxicity against MCF-7 (Michigan Cancer Foundation-7) cells in vitro by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. At an extract concentration of 100 μg/mL, 4 isolates out of 12 showed high activity against the cancer cell growth. The four isolates were then selected for further IC50 determination, by measuring the inhibition of cancer cell proliferation at extract concentration of 25 μg/mL, 50 μg/mL, 100 μg/mL, 200 μg/mL and 400 μg/mL. Results showed that isolate Sir-G5 had the highest anticancer activity with an IC50 of 19.20 μg/mL. The best isolates were screened again using a normal cell (Chang cells) to determine its toxicity against normal cells. Results indicated that the extracts do not affect the proliferation of normal cells. Molecular identification showed that the fungal isolate Sir-G5 has a close relationship with Phomopsis sp. The endophytic fungi isolated from soursop leaf has the potential to be used as a source of anticancer agents. Copyright © 2017 Hainan Medical University. Production and hosting by Elsevier B.V. All rights reserved.

  16. An endophytic Streptomyces sp. strain DHV3-2 from diseased root as a potential biocontrol agent against Verticillium dahliae and growth elicitor in tomato (Solanum lycopersicum).

    PubMed

    Cao, Peng; Liu, Chongxi; Sun, Pengyu; Fu, Xuepeng; Wang, Shaoxian; Wu, Fengzhi; Wang, Xiangjing

    2016-12-01

    Plant endophytes play important roles in biocontrol of plant diseases. Actinomycetes are used for biocontrol of fungal diseases caused by Verticillium dahliae. Many studies have focused on the endophytic actinomycetes isolated from the roots of healthy plants, but few on those from the roots of diseased plants. In the present research, actinomycetes were isolated from the roots of diseased and healthy tomato plants, respectively. The results showed that, in total, 86 endophytic actinomycetes were isolated for screening of their antimicrobial activities, 8 of which showed antagonism to V. dahliae in vitro. Among the 8 antagonistic strains, 5 (out of 36) were from the roots of diseased plants, with inhibition diameter zones ranging from 11.2 to 18.2 mm, whereas 3 (out of 50) were from the roots of healthy plants, with inhibition diameter zones ranging from 11.5 to 15.5 mm. Endophytic strain DHV3-2 was isolated from the root of a diseased plant and demonstrated a potent effect against V. dahliae and other pathogenic fungi by showing the largest inhibition diameter zones among all the eight antagonistic strains. Thus, strain DHV3-2 was chosen to investigate its biological control efficacies in vivo. Further study showed that the disease incidence and disease severity indices of tomato Verticillium wilt decreased significantly (P < 0.05). We also found that the plant shoot fresh weight and height increased greatly (P < 0.05) upon treatment with strain DHV3-2 compared to the plants uninoculated in greenhouse conditions. Root colonization showed that strain DHV3-2 had the higher root-colonizing capacity in the roots of infected plants compared with the roots of healthy plants. This isolate was identified as Streptomyces sp. based on morphological characteristics and 16S rRNA gene analysis. In conclusion, the roots of diseased tomato plants are a potential reservoir of biological control actinomycetes, and Streptomyces sp. strain DHV3-2 is a potential biocontrol

  17. Effects of a dark-septate endophytic isolate LBF-2 on the medicinal plant Lycium barbarum L.

    PubMed

    Zhang, Hai-Han; Tang, Ming; Chen, Hui; Wang, Ya-Jun

    2012-02-01

    Dark septate endophytes (DSE) are ubiquitous root associated fungi; however, our understanding of their ecological function remains unclear. Here, we investigated the positive effect of a DSE fungus on its host plant Lycium barbarum L. A DSE isolate, LBF-2, isolated from the roots of L. barbarum, was inoculated onto the roots of plants, which were grown under greenhouse conditions for five weeks. The result of molecular analyses of internal transcribed spacer regions indicated that LBF-2 was 96% similar to Paraphoma chrysanthemicola. Melanized septate hyphae were observed in the root cortical cells of L. barbarum using a light microscope. Inoculation with LBF-2 increased the total biomass by 39.2% and also enhanced chlorophyll fluorescence. Inoculation increased the concentration of total chlorophyll by 22.8% and of chlorophyll a by 21.3%, relative to uninoculated controls. These data indicate that the LBF-2 isolate might be used to facilitate the cultivation of L. barbarum, which has medicinal applications.

  18. Description of Kibdelosporangium banguiense sp. nov., a novel actinomycete isolated from soil of the forest of Pama, on the plateau of Bangui, Central African Republic.

    PubMed

    Pascual, Javier; González, Ignacio; Estévez, Mar; Benito, Patricia; Trujillo, Martha E; Genilloud, Olga

    2016-05-01

    A novel actinomycete strain F-240,109(T) from the MEDINA collection was isolated from a soil sample collected in the forest of Pama, on the plateau of Bangui, Central African Republic. The strain was identified according to its 16S rRNA gene sequence as a new member of the genus Kibdelosporangium, being closely related to Kibdelosporangium aridum subsp. aridum (98.6 % sequence similarity), Kibledosporangium phytohabitans (98.3 %), Kibdelosporangium aridum subsp. largum (97.7 %), Kibdelosporangium philippinense (97.6 %) and Kibledosporangium lantanae (96.9 %). In order to resolve its precise taxonomic status, the strain was characterised through a polyphasic approach. The strain is a Gram-stain positive, aerobic, non-motile and catalase-positive actinomycete characterised by formation of extensively branched substrate mycelia and sparse brownish grey aerial mycelia with sporangium-like globular structures. The chemotaxonomic characterisation of strain F-240,109(T) corroborated its affiliation into the genus Kibdelosporangium. The peptidoglycan contains meso-diaminopimelic acid; the major menaquinone is MK-9(H4); the phospholipid profile contains high amounts of phosphatidylethanolamine, hydroxyphosphatidylethanolamine, diphosphatidylglycerol and an unidentified phospholipid; and the predominant cellular fatty acid methyl esters are iso-C16:0, iso-C14:0, iso-C15:0 and 2OH iso-C16:0. However, some key phenotypic differences regarding to its close relatives and DNA-DNA hybridization values indicate that strain F-240,109(T) represents a novel Kibdelosporangium species, for which the name Kibdelosporangium banguiense sp. nov. is proposed. The type strain is strain F-240,109(T) (=DSM 46670(T), =LMG 28181(T)).

  19. Comparison of the chemistry and diversity of endophytes isolated from wild-harvested and greenhouse-cultivated yerba mansa (Anemopsis californica)

    PubMed Central

    Bussey, Robert O.; Kaur, Amninder; Todd, Daniel A.; Egan, Joseph M.; El-Elimat, Tamam; Graf, Tyler N.; Raja, Huzefa A.; Oberlies, Nicholas H.; Cech, Nadja B.

    2015-01-01

    With this study, we explored the identity and chemistry of fungal endophytes from the roots of yerba mansa [Anemopsis californica (Nutt.) Hook. & Arn. (Saururaceae)], a botanical traditionally used to treat infection. We compared the diversity of fungal endophytes isolated from a wild-harvested A. californica population, and those from plants cultivated for one year in a greenhouse environment. The wild-harvested population yielded thirteen fungal strains (eleven unique genotypes). Of the extracts prepared from these fungi, four inhibited growth of Staphylococcus aureus by >25% at 20 µg/mL, and three inhibited growth of Pseudomonas aeruginosa by ≥20% at 200 µg/mL. By comparison, A. californica roots after one year of cultivation in the greenhouse produced only two unique genotypes, neither of which displayed significant antimicrobial activity. The fungus Chaetomium cupreum isolated from wild-harvested A. californica yielded a new antimicrobial spirolactone, chaetocuprum (1). An additional fourteen known compounds were identified using LC-MS dereplication of the various fungal endophytes. This study provides new insights into the identity and chemistry of A. californica fungal endophytes, and demonstrates the importance of considering growing conditions when pursuing natural product drug discovery from endophytic fungi. PMID:25642298

  20. Pseudomonas tarimensis sp. nov., an endophytic bacteria isolated from Populus euphratica.

    PubMed

    Anwar, Nusratgul; Rozahon, Manziram; Zayadan, Bolatkhan; Mamtimin, Hormathan; Abdurahman, Mehfuzem; Kurban, Marygul; Abdurusul, Mihribangul; Mamtimin, Tursunay; Abdukerim, Muhtar; Rahman, Erkin

    2017-10-06

    An endophytic bacterium, MA-69(T), was isolated from the storage liquid in the stems of Populuseuphratica trees at the ancient Ugan River in Xinjiang, PR China. Strain MA-69(T) was found to be short rod-shaped, Gram-stain-negative, non-spore-forming, aerobic and motile by means of a monopolar flagellum. According to phylogenetic analysis based on 16S rRNA gene sequences, strain MA-69(T) was assigned to the genus Pseudomonas with highest 16S rRNA gene sequence similarity of 97.5 % to Pseudomonas azotifigens JCM 12708(T), followed by Pseudomonas matsuisoli JCM 30078(T) (97.5 %), Pseudomonas balearica DSM 6083(T) (97.1 %), Azotobacter salinestris ATCC 49674(T) (96.1 %) and Pseudomonas indica DSM 14015(T) (95.9 %). Analysis of strain MA-69(T) based on the three housekeeping genes, rpoB, rpoD and gyrB, further confirmed the isolate to be distinctly delineated from species of the genus Pseudomonas. The DNA G+C content of strain MA-69(T) was 64.1 mol%. DNA-DNA hybridization with Pseudomonas azotifigens JCM 12708(T), Pseudomonas matsuisoli JCM 30078(T) and Pseudomonas balearica DSM 6083(T) revealed 62.9, 60.1 and 49.0 % relatedness, respectively. The major fatty acids in strain MA-69(T) were summed feature 3 (25.7 %), summed feature 8 (24.0 %), C19 : 0cyclo ω8c (19.9 %), C16 : 0 (14.6 %) and C12 : 0 (6.3 %). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Q-9 was the major quinone in strain MA-69(T). Based on phenotypic, chemotaxonomic and phylogenetic properties, strain MA-69(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas tarimensis sp. nov. is proposed. The type strain is MA-69(T) (=CCTCC AB 2013065(T)=KCTC 42447(T)).

  1. Application of actinomycetes to soil to ameliorate water repellency.

    PubMed

    McKenna, F; El-Tarabily, K A; Petrie, S; Chen, C; Dell, B

    2002-01-01

    The aim of this study was to develop a novel isolation technique using a mixture of Bacillus and Streptomyces phages to selectively isolate wax-utilizing non-streptomycete actinomycetes effective in ameliorating water repellency in a problem soil. Phages added to a soil suspension reduced the dominance of Bacillus and Streptomyces isolates and significantly increased the number of non-streptomycete actinomycetes on isolation plates. Promising isolates, grown on a medium containing beeswax as sole carbon source, were selected for application to water repellent soil. Their addition significantly reduced water repellency. Phage application significantly increased the isolation of non-streptomycete actinomycetes. Wax-utilizing isolates were found to significantly reduce water repellency in a problem soil. The phage technique can be used for the routine isolation of non-streptomycete actinomycetes. Beeswax medium can be used to selectively isolate wax-utilizing micro-organisms with the potential to ameliorate water repellency in soil.

  2. Biosynthesis and genomic analysis of medium-chain hydrocarbon production by the endophytic fungal isolate Nigrograna mackinonnii E5202H

    PubMed Central

    Shaw, Jeffery J; Spakowicz, Daniel J; Dalal, Rahul S; Davis, Jared H; Lehr, Nina A; Dunican, Brian F; Orellana, Esteban A; Narváez-Trujillo, Alexandra; Strobel, Scott A

    2015-01-01

    An endophytic fungus was isolated that produces a series of volatile natural products, including terpenes and odd chain polyenes. Phylogenetic analysis of the isolate using five loci suggests that it is closely related to Nigrograna mackinnonii CBS 674.75. The main component of the polyene series was purified and identified as (3E,5E,7E)-nona-1,3,5,7-tetraene (NTE), a novel natural product. Non-oxygenated hydrocarbons of this chain length are uncommon and desirable as gasoline-surrogate biofuels. The biosynthetic pathway for NTE production was explored using metabolic labeling and GCMS. Two-carbon incorporation 13C acetate suggests that it is derived from a polyketide synthase (PKS) followed by decarboxylation. There are several known mechanisms for such decarboxylation, though none have been discovered in fungi. Towards identifying the PKS responsible for the production of NTE, the genome of N. mackinnonii E5202H (ATCC SD-6839) was sequenced and assembled. Of the 32 PKSs present in the genome, 17 are predicted to contain sufficient domains for the production of NTE. These results exemplify the capacity of endophytic fungi to produce novel natural products that may have many uses, such as biologically derived fuels and commodity chemicals. PMID:25672844

  3. An actinomycete isolate from solitary wasp mud nest having strong antibacterial activity and kills the Candida cells due to the shrinkage and the cytosolic loss

    PubMed Central

    Kumar, Vijay; Naik, Bindu; Gusain, Omprakash; Bisht, Gajraj S.

    2014-01-01

    An actinomycetes strain designated as MN 2(6) was isolated from the solitary wasp mud nest. The isolate was identified using polyphasic taxonomy. It produced the extensive branched brown substrate and white aerial hyphae that changed into grayish black. The aerial mycelia produced the spiral spore chains with rugose spore surface. The growth was observed between temperature range of 27–37°C, pH 8–10 and below salt concentration of 6% (w/v). The comparative analysis of 16S rRNA gene sequence and phylogenetic relationship showed that strain MN 2(6) lies in clade with Streptomyces hygroscopicus subsp. hygroscopicus NRRL 2387T, Streptomyces sporocinereus NBRC 100766T and Streptomyces demainii NRRL B-1478T with which it shares a 16S rRNA gene sequence similarity of 99.3%. The strain MN 2(6) can be differentiated from type strains based on phenotypic characteristics. The strain MN 2(6) showed most promising activity against Gram-positive, Gram-negative bacteria, acid-fast bacilli and Candida species suggesting broad-spectrum characteristics of the active metabolite. Evaluation of anti-candidal activity of the metabolite of strain MN 2(6) by scanning electron microscopy (SEM) revealed changed external morphology of yeast. It kills the Candida cells due to the shrinkage and the cytosolic loss. However, further studies are required to elucidate the structure of the active metabolite produced by the isolate MN 2(6). PMID:25191320

  4. Actinomycetes in relation to taste and odour in drinking water: myths, tenets and truths.

    PubMed

    Zaitlin, Beryl; Watson, Susan B

    2006-05-01

    Actinomycetes are a complex group of bacteria present in a wide variety of environments, either as dormant spores or actively growing. Some actinomycetes produce two potent terpenoids (geosmin and 2-methylisoborneol (MIB)) and pyrazines, common causes of drinking water off flavours, and have been implicated in taste and odour episodes. However, isolation from a water source is not evidence that actinomycetes caused a taste and odour event. Dormant spores of actinomycetes may be isolated from aquatic environments in high concentrations, despite production in the terrestrial environment. Similarly, odourous compounds produced by actinomycetes may be produced terrestrially and washed into aquatic environments, with or without the actinomycetes that produced them. Actinomycetes may exist as actively growing mycelium in small, specialized habitats within an aquatic system, but their odourous compounds may influence a wider area. This paper attempts to elucidate the types and activities of actinomycetes that may be found in, or interact with, drinking water supplies.

  5. Molecular characterization by amplified ribosomal DNA restriction analysis and antimicrobial potential of endophytic fungi isolated from Luehea divaricata (Malvaceae) against plant pathogenic fungi and pathogenic bacteria.

    PubMed

    Bernardi-Wenzel, J; Garcia, A; Azevedo, J L; Pamphile, J A

    2013-10-29

    Luehea divaricata is an important plant in popular medicine; it is used for its depurative, anti-inflammatory, and other therapeutic activities. We evaluated the antimicrobial activity of endophytic fungi isolated from leaves of L. divaricata against phytopathogens and pathogenic bacteria, and characterized the isolates based on amplified ribosomal DNA restriction analysis (ARDRA). The in vitro antagonistic activity of these endophytes against the phytopathogen Alternaria alternata was assayed by dual culture technique. Based on this evaluation of antimicrobial activity, we extracted secondary metabolites from nine endophytic fungi by partitioning in ethyl acetate and methanol. These were tested against the phytopathogens A. alternata, Colletotrichum sp and Moniliophthora perniciosa, and against the human pathogenic bacteria Escherichia coli and Staphylococcus aureus. Molecular characterization by ARDRA technique was used for phylogenetic analysis, based on comparison with sequences in GenBank. The endophytes had varied effects on A. alternata. One isolate produced an inhibition halo against M. perniciosa and against E. coli. This antibiosis activity indicates a role in the protection of the plant against microbial pathogens in nature, with potential for pharmaceutical and agricultural applications. Based on ARDRA, the 13 isolates were grouped. We found three different haplotypes of Phomopsis sp, showing interspecific variability. It appears that examination of the microbial community associated with medicinal plants of tropical regions has potential as a useful strategy to look for species with biotechnological applications.

  6. Draft Genome Sequence of Bacillus pumilus Strain GM3FR, an Endophyte Isolated from Aerial Plant Tissues of Festuca rubra L.

    PubMed Central

    Hollensteiner, Jacqueline; Daniel, Rolf; Liesegang, Heiko; Vidal, Stefan

    2017-01-01

    ABSTRACT Here, we report the draft genome sequence of Bacillus pumilus GM3FR, an endophytic bacterium isolated from aerial plant tissues of Festuca rubra L. The draft genome consists of 3.5 Mb and harbors 3,551 predicted protein-encoding genes. The genome provides insights into the biocontrol potential of B. pumilus GM3FR. PMID:28360161

  7. Draft Genome Sequence of Curtobacterium sp. Strain ER1/6, an Endophytic Strain Isolated from Citrus sinensis with Potential To Be Used as a Biocontrol Agent.

    PubMed

    Garrido, Leandro Maza; Alves, João Marcelo Pereira; Oliveira, Liliane Santana; Gruber, Arthur; Padilla, Gabriel; Araújo, Welington Luiz

    2016-11-17

    Herein, we report a draft genome sequence of the endophytic Curtobacterium sp. strain ER1/6, isolated from a surface-sterilized Citrus sinensis branch, and it presented the capability to control phytopathogens. Functional annotation of the ~3.4-Mb genome revealed 3,100 protein-coding genes, with many products related to known ecological and biotechnological aspects of this bacterium.

  8. Rhizobium populi sp. nov., an endophytic bacterium isolated from Populus euphratica.

    PubMed

    Rozahon, Manziram; Ismayil, Nurimangul; Hamood, Buayshem; Erkin, Raziya; Abdurahman, Mehfuzem; Mamtimin, Hormathan; Abdukerim, Muhtar; Lal, Rup; Rahman, Erkin

    2014-09-01

    An endophytic bacterium, designated K-38(T), was isolated from the storage liquid in the stems of Populus euphratica trees at the ancient Ugan River in Xinjiang, PR China. Strain K-38(T) was found to be rod-shaped, Gram-stain-negative, aerobic, non-motile and non-spore-forming. Strain K-38(T) grew at temperatures of 25-37 °C (optimum, 28 °C), at pH 6.0-9.0 (optimum, pH 7.5) and in the presence of 0-3 % (w/v) NaCl with 1 % as the optimum concentration for growth. According to phylogenetic analysis based on 16S rRNA gene sequences, strain K-38(T) was assigned to the genus Rhizobium with highest 16S rRNA gene sequence similarity of 97.2 % to Rhizobium rosettiformans W3(T), followed by Rhizobium nepotum 39/7(T) (96.5 %) and Rhizobium borbori DN316(T) (96.2 %). Phylogenetic analysis of strain K-38(T) based on the protein coding genes recA, atpD and nifH confirmed (similarities were less than 90 %) it to be a representative of a distinctly delineated species of the genus Rhizobium. The DNA G+C content was determined to be 63.5 mol%. DNA-DNA relatedness between K-38(T) and R. rosettiformans W3(T) was 48.4 %, indicating genetic separation of strain K-38(T) from the latter strain. The major components of the cellular fatty acids in strain K-38(T) were revealed to be summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c; 57.2 %), C16 : 0 (13.6 %) and summed feature 2 (comprising C12 : 0 aldehyde, C14 : 0 3-OH/iso-C16 : 1 I and/or unknown ECL 10.928; 11.0 %). Polar lipids of strain K-38(T) include phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, two unidentified aminophospholipids and two unidentified phospholipids. Q-10 was the major quinone in strain K-38(T). Based on phenotypic, chemotaxonomic and phylogenetic properties, strain K-38(T) represents a novel species of the genus Rhizobium, for which the name Rhizobium populi sp. nov. is proposed

  9. Larsenimonas suaedae sp. nov., a moderately halophilic, endophytic bacterium isolated from the euhalophyte Suaeda salsa.

    PubMed

    Xia, Zhi-Jie; Wu, Hong-Zhen; Cui, Chun-Xiao; Chen, Qi; Zhao, Guo-Yan; Wang, Hai-Xia; Dai, Mei-Xue

    2016-08-01

    A moderately halophilic, Gram-stain-negative, non-endospore-forming endophytic bacterium designated strain ST307T was isolated from the euhalophyte Suaeda salsa in Dongying, China. Strain ST307T was aerobic, rod-shaped, motile and orange-yellow-pigmented. The organism grew at NaCl concentrations of 0.6-20 % (w/v) (optimum 5-6 %, w/v), at temperatures of 5-45 °C (optimum 35 °C) and at pH 5-9 (optimum pH 7-8). It accumulated poly-β-hydroxybutyric acid and produced exopolysaccharides. The major fatty acids were C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C16 : 1ω7c/C16 : 1ω6c. The predominant lipoquinone was ubiquinone Q-9. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, a glycoaminolipid and a phosphoglycoaminolipid. The DNA G+C content was 60.5 mol%. Phylogenetic analyses of 16S rRNA gene sequences and concatenated atpA, rpoD and secA gene sequences revealed that the strain represents a member of the genus Larsenimonas. The closest related type strain was Larsenimonas salina M1-18T. Mean DNA-DNA relatedness values between strain ST307T and the related species L. salina M1-18T, Chromohalobacter beijerinckii DSM 7218T, C. canadensis DSM 6769T, C. israelensis DSM 6768T, C. marismortui CGMCC 1.2321T, C. nigrandesensis DSM 14323T, C. salexigens DSM 3043T and C. sarecensis DSM 15547T were 15±2-45±1 %. On the basis of phenotypic, chemotaxonomic and molecular features, strain ST307T clearly represents a novel species of the genus Larsenimonas. The name Larsenimonassuaedae sp. nov. is proposed, with ST307T (=CGMCC 1.8902T=DSM 22428T) as the type strain.

  10. Mangrovibacter phragmitis sp. nov., an endophyte isolated from the roots of Phragmites karka.

    PubMed

    Behera, Pratiksha; Venkata Ramana, V; Maharana, Bhagirathi; Joseph, Neetha; Vaishampayan, Parag; Singh, Nitin K; Shouche, Yogesh; Bhadury, Punyasloke; Mishra, Samir R; Raina, Vishakha; Suar, Mrutyunjay; Pattnaik, Ajit K; Rastogi, Gurdeep

    2017-05-01

    A facultatively anaerobic, Gram-stain-negative, rod-shaped, nitrogen-fixing, endophytic bacterial strain designated MP23T was isolated from the roots of Phragmites karka growing in Chilika Lagoon, Odisha, India. Strain MP23T was slightly halophilic, and the optimal NaCl concentration and temperature for growth were 1 % and 30 °C, respectively. On the basis of 16S rRNA gene sequence similarities, strain MP23T was affiliated to the family Enterobacteriaceae and most closely related to Mangrovibacter yixingensis KCTC 42181T and Mangrovibacter plantisponsor DSM 19579T with 99.71 % similarity, followed by Salmonella enterica subsp. salamae DSM 9220T (97.22 %), Cronobacter condimenti LMG 26250T (97.14 %) and Salmonella enterica subsp. diarizonae DSM 14847T (97 %). Sequence analysis of 16S rRNA, hsp60, gyrB and rpoB genes showed that strain MP23T formed a phylogenetic cluster with M. yixingensis KCTC 42181T and M. plantisponsor DSM 19579T indicating that it belongs to the genus Mangrovibacter. The major cellular fatty acids were C16 : 0, C18 : 1ω6c and/or C18 : 1ω7c, C16 : 1ω6c and/or C16 : 1ω7c, C14 : 0, C14 : 0 3-OH and/or iso-C16 : 1 I and C17 : 0 cyclo. Polar lipids of strain MP23T consisted of phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 50.3 mol%. Based on experimental DNA-DNA hybridization values and average nucleotide identity derived from in silico comparison of whole-genome sequences, strain MP23T could be distinguished from its closest neighbours. We therefore conclude that strain MP23T represents a novel species of the genus Mangrovibacter for which the name Mangrovibacter phragmitis sp. nov. is proposed. The type strain is MP23T (=DSM 100250T=KCTC 42580T).

  11. Antibacterial agents from actinomycetes - a review.

    PubMed

    Mahajan, Girish Badrinath; Balachandran, Lakshmi

    2012-01-01

    The discovery of Penicillin in 1928 and that of Streptomycin in 1943, has been pivotal to the exploration of nature as a source of new lead molecules. Globally, the microbiologist today is acknowledged as a crucial player in the drug discovery program. The microbial products, especially those from actinomycetes have been a phenomenal success for the past seven decades. Bioprospecting for new leads are often compounded by the recurrence of known antibiotics in newer microbial isolates. Despite all these deterrents, actinomycetes have proved to be a sustained mine of novel antibiotics, which selectively destroys the pathogens without affecting the host tissues. Each of these antibiotics is unique in their mode of action. Their versatility and immense economic value is something, which is extremely noteworthy. The anti-infective turn-over of over 79 billion US dollars in 2009, includes about 166 antibiotics and derivatives such as the Beta-lactam peptide antibiotics, the macrolide polyketide erythromycin, tetracyclines, aminoglycosides, daptomycin, tigecycline, most of which are produced by actinomycetes (1). Actinomycetes continue to play a highly significant role in drug discovery and development. Among the bioactive compounds that have been obtained so far from microbes, 45 % are produced by actinomycetes, 38 % by fungi and 17 % by unicellular eubacteria (2). Further many chemically synthesized drugs owe their origin to natural sources. In this review article, we highlight the recent antibiotics from actinomycetes with emphasis on their source, structures, activity, mechanism of action and current status.

  12. Pyrrole and indole alkaloids from an endophytic Fusarium incarnatum (HKI00504) isolated from the mangrove plant Aegiceras corniculatum.

    PubMed

    Li, Li-Ya; Ding, Yi; Groth, Ingrid; Menzel, Klaus-Dieter; Peschel, Gundela; Voigt, Kerstin; Deng, Zi-Wei; Sattler, Isabel; Lin, Wen-Han

    2008-01-01

    Two new pyrrole alkaloids, N-[4-(2-formyl-5-hydroxymethyl-pyrrol-1-yl)-butyl]-acetamide (1) and N-[5-(2-formyl-5-hydroxymethyl-pyrrol-1-yl)-pentyl]-acetamide (2), and a new indole derivative (3aR,8aR)-3a-acetoxyl-1,2,3,3a,8,8a-hexahydropyrrolo-[2,3-b]indol (3) were isolated, together with ( - )-3a-hydroxyfuroindoline, (3aR,8aS)-1-acetyl-1,3,3a,8,8a-hexahydropyrrolo-[2,3-b]indol-3a-ol, and N-acetyltryptamine A, from an endophytic ascomycetous fungus, Fusarium incarnatum (HKI00504), which was isolated from the mangrove plant Aegiceras corniculatum. The structures of compounds 1-3 were determined on the basis of extensive spectroscopic data analyses.

  13. Isolation and identification of endophytic fungi in roots of nine Holcoglossum plants (Orchidaceae) collected from Yunnan, Guangxi, and Hainan provinces of China.

    PubMed

    Tan, Xiao-Ming; Chen, Xiao-Mei; Wang, Chun-Lan; Jin, Xiao-Hua; Cui, Jin-Long; Chen, Juan; Guo, Shun-Xing; Zhao, Li-Fang

    2012-02-01

    Holcoglossum is one of the smaller genera of Orchidaceae, mainly distributed in southwest China. Some members of this genus as well as H. rupestre and H. flavescens are endemic and rare Chinese orchids. As far as we know, little work has been done concerning the relationships between the Holcoglossum plants and endophytic microorganisms. In this study, 46 culturable fungal endophytes were isolated and identified from roots of nine Holcoglossum plants collected from Yunnan, Guangxi, and Hainan provinces of China based on molecular techniques. The results showed that all strains belonged to four classes, i.e., Sordariomycetes (41.30%), Dothideomycetes (36.96%), Agaricomycetes (17.39%), Leotiomycetes (4.35%). Thirty-six strains were identified at the genus level, including Alternaria, Cladosporium, Clonostachys, Colletotrichum, Cosmospora, Cryptosporiopsis, Cylindrocarpon, Didymella, Epulorhiza (Anamorphic Tulasnella), Fusarium, Myrmecridium, Leptosphaeria, Paraconiothyrium, Phomopsis, Pyrenochaeta, and Stephanonectria. Fusarium and Epulorhiza (Anamorphic Tulasnella) were the dominant fungal endophytes. Some orchids mycorrhizal fungi as well as Tulasnella calospora and Epulorhiza sp. were found in roots. This is the first report concerning endophytic fungi from Holcoglossum plants (Orchidaceae), suggesting that endophytic fungi in Holcoglossum plants are very abundant.

  14. Streptomyces oceani sp. nov., a new obligate marine actinomycete isolated from a deep-sea sample of seep authigenic carbonate nodule in South China Sea.

    PubMed

    Tian, Xin-Peng; Xu, Ying; Zhang, Jing; Li, Jie; Chen, Zhong; Kim, Chang-Jin; Li, Wen-Jun; Zhang, Chang-Sheng; Zhang, Si

    2012-08-01

    A novel aerobic actinomycete strain, designated as SCSIO 02100(T), was isolated from a deep sea sediment sample collected from Northern South China Sea at a depth of 578 m. This isolate requires sea water or a sodium-supplemented medium for growth. BLAST searches based on the almost full length of the 16S rRNA gene sequence, showed that strain SCSIO 02100(T) had the highest similarities with Streptomyces armeniacus (JCM 3070(T)) (97.1 %). Phylogenetic trees reconstructed on the basis of 16S rRNA gene sequences revealed that strain SCSIO 02100(T) formed a distinct lineage with S. nanshensis SCSIO 01066(T) with 96.9 % similarity. Further analysis of the polyphasic taxonomic data, including morphological, phenotypic and chemotaxonomic properties, showed that strain SCSIO 02100(T) could be readily distinguished from the most closely related members of the genus Streptomyces. Thus, based on the polyphasic taxonomic data, a novel species, Streptomyces oceani sp. nov., is proposed, with the type strain SCSIO 02100(T) (=DSM 42043(T) = CGMCC 4.7007(T)).

  15. Isolation, plant colonization potential, and phenanthrene degradation performance of the endophytic bacterium Pseudomonas sp. Ph6-gfp

    NASA Astrophysics Data System (ADS)

    Sun, Kai; Liu, Juan; Gao, Yanzheng; Jin, Li; Gu, Yujun; Wang, Wanqing

    2014-06-01

    This investigation provides a novel method of endophyte-aided removal of polycyclic aromatic hydrocarbons (PAHs) from plant bodies. A phenanthrene-degrading endophytic bacterium Pseudomonas sp. Ph6 was isolated from clover (Trifolium pratense L.) grown in a PAH-contaminated site. After being marked with the GFP gene, the colonization and distribution of strain Ph6-gfp was directly visualized in plant roots, stems, and leaves for the first time. After ryegrass (Lolium multiflorum Lam.) roots inoculation, strain Ph6-gfp actively and internally colonized plant roots and transferred vertically to the shoots. Ph6-gfp had a natural capacity to cope with phenanthrene in vitro and in planta. Ph6-gfp degraded 81.1% of phenanthrene (50 mg.L-1) in a culture solution within 15 days. The inoculation of plants with Ph6-gfp reduced the risks associated with plant phenanthrene contamination based on observations of decreased concentration, accumulation, and translocation factors of phenanthrene in ryegrass. Our results will have important ramifications in the assessment of the environmental risks of PAHs and in finding ways to circumvent plant PAH contamination.

  16. Isolation, plant colonization potential, and phenanthrene degradation performance of the endophytic bacterium Pseudomonas sp. Ph6-gfp

    PubMed Central

    Sun, Kai; Liu, Juan; Gao, Yanzheng; Jin, Li; Gu, Yujun; Wang, Wanqing

    2014-01-01

    This investigation provides a novel method of endophyte-aided removal of polycyclic aromatic hydrocarbons (PAHs) from plant bodies. A phenanthrene-degrading endophytic bacterium Pseudomonas sp. Ph6 was isolated from clover (Trifolium pratense L.) grown in a PAH-contaminated site. After being marked with the GFP gene, the colonization and distribution of strain Ph6-gfp was directly visualized in plant roots, stems, and leaves for the first time. After ryegrass (Lolium multiflorum Lam.) roots inoculation, strain Ph6-gfp actively and internally colonized plant roots and transferred vertically to the shoots. Ph6-gfp had a natural capacity to cope with phenanthrene in vitro and in planta. Ph6-gfp degraded 81.1% of phenanthrene (50 mg·L−1) in a culture solution within 15 days. The inoculation of plants with Ph6-gfp reduced the risks associated with plant phenanthrene contamination based on observations of decreased concentration, accumulation, and translocation factors of phenanthrene in ryegrass. Our results will have important ramifications in the assessment of the environmental risks of PAHs and in finding ways to circumvent plant PAH contamination. PMID:24964867

  17. Biodegradation of phenol and benzene by endophytic bacterial strains isolated from refinery wastewater-fed Cannabis sativa.

    PubMed

    Iqbal, Aneela; Arshad, Muhammad; Hashmi, Imran; Karthikeyan, Raghupathy; Gentry, Terry J; Schwab, Arthur Paul

    2017-06-13

    The presence of benzene and phenol in the environment can lead to serious health effects in humans and warrant development of efficient cleanup strategies. The aim of the present work was to assess the potential of indigenous endophytic bacterial strains to degrade benzene and phenol. Seven strains were successfully isolated from Cannabis sativa plants irrigated with oil refinery wastewater. Molecular characterization was performed by 16S rRNA gene sequencing. Phenol was biodegraded almost completely with Achromobacter sp. (AIEB-7), Pseudomonas sp. (AIEB-4), and Alcaligenes sp. (AIEB-6) at 250, 500, and 750 mg L(-1); however, the degradation was only 81%, 72%, and 69%, respectively, when exposed to 1000 mg L(-1). Bacillus sp. (AIEB-1), Enterobacter sp. (AIEB-3), and Acinetobacter sp. (AIEB-2) degraded benzene significantly at 250, 500, and 750 mg L(-1). However, these strains showed 80%, 72%, and 68% benzene removal at 1000 mg L(-1) exposure, respectively. Rates of degradation could be modeled with first-order kinetics with rate constant values of 1.86 × 10(-2) for Pseudomonas sp. (AIEB-4) and 1.80 × 10(-2) h(-1) for Bacillus sp. (AIEB-1) and half-lives of 1.5 and 1.6 days, respectively. These results establish a foundation for further testing of the phytoremediation of hydrocarbon-contaminated soils in the presence of these endophytic bacteria.

  18. Isolation, plant colonization potential, and phenanthrene degradation performance of the endophytic bacterium Pseudomonas sp. Ph6-gfp.

    PubMed

    Sun, Kai; Liu, Juan; Gao, Yanzheng; Jin, Li; Gu, Yujun; Wang, Wanqing

    2014-06-26

    This investigation provides a novel method of endophyte-aided removal of polycyclic aromatic hydrocarbons (PAHs) from plant bodies. A phenanthrene-degrading endophytic bacterium Pseudomonas sp. Ph6 was isolated from clover (Trifolium pratense L.) grown in a PAH-contaminated site. After being marked with the GFP gene, the colonization and distribution of strain Ph6-gfp was directly visualized in plant roots, stems, and leaves for the first time. After ryegrass (Lolium multiflorum Lam.) roots inoculation, strain Ph6-gfp actively and internally colonized plant roots and transferred vertically to the shoots. Ph6-gfp had a natural capacity to cope with phenanthrene in vitro and in planta. Ph6-gfp degraded 81.1% of phenanthrene (50 mg · L(-1)) in a culture solution within 15 days. The inoculation of plants with Ph6-gfp reduced the risks associated with plant phenanthrene contamination based on observations of decreased concentration, accumulation, and translocation factors of phenanthrene in ryegrass. Our results will have important ramifications in the assessment of the environmental risks of PAHs and in finding ways to circumvent plant PAH contamination.

  19. Chemical Assessment and Antimicrobial and Antioxidant Activities of Endophytic Fungi Extracts Isolated from Costus spiralis (Jacq.) Roscoe (Costaceae)

    PubMed Central

    Marson Ascêncio, Poliana Guerino; Ascêncio, Sérgio Donizeti; Aguiar, Aline Aires; Fiorini, Adriana; Pimenta, Raphael Sanzio

    2014-01-01

    Costus spiralis (Costaceae) is a species native to the Amazon region and is used in traditional medicine. The endophytic fungi used in this study were obtained from leaves of this plant. 13 strains were selected to obtain hydroethanolic extracts and were submitted to hydroalcoholic extraction and evaluated for antioxidant activity by DPPH (2,2-difenil-1-picrilhidrazil) and FRAP (ferric reducing antioxidant power), and all of the fungi had positive results. The antimicrobial action of crude extracts had a good range of activities. All extracts had inhibitory activities against the yeasts of Candida albicans and C. parapsilosis, with 125 to 500 μg/mL MIC. Eight extracts had antimicrobial activities against Bacillus subtilis (MIC from 62.4 to 125 μg/mL), 5 against Pseudomonas aeruginosa (MIC from 125 to 500 μg/mL), 2 against Salmonella enterica (MIC from 125 to 62.5 μg/mL), and 2 against Enterococcus faecalis (MIC from 500 to 125 μg/mL). The presence of secondary metabolites, including coumarins, was observed during chemical evaluation by thin layer chromatography. Total phenol content was estimated, and a strong positive correlation to antioxidant activity was observed, according to its Pearson coefficient. This is the first report of the bioactive potential of endophytic fungi isolated from the Costaceae family in Brazilian ecosystems. PMID:25587339

  20. Assessment of genetic diversity and distribution of endophytic fungal communities of Alternaria solani isolates associated with the dominant Karanja plants in Sanganer Region of Rajasthan.

    PubMed

    Tiwari, Kartikeya; Chittora, Manish

    2013-12-01

    Higher plants are ubiquitously colonized with fungal endophytes that often lack readily detectable structures. Current study examines the distribution of endophytic fungal communities within Karanja plants and diversity of novel fungal endophyte Alternaria solani isolates collected from different locations of Sanganer region of Rajasthan. Results confirmed that A. solani is a major fungal endophyte consortium associated with Karanja plants. PCR Amplified fragments using random amplified polymorphic DNA (RAPD) primers were subjected to unweighted pair group method analysis (UPGMA), which clearly distinguished twelve ecologically diverse A. solani isolates. A total of 58 RAPD loci were amplified, out of which 35 (60.34%) were polymorphic and 23 were monomorphic (39.66%) in nature. These polymorphic loci were identified with an average of 2.92 bands per primer. The efficacy of RAPD markers proved as an efficient marker system with respect to detection of polymorphism and number of loci scored and can be used for the identification of a particular isolates, thereby defining core collections and strengthening their exploitation in acquiring novel products produced by them.

  1. Isolation and characterization of Stemphylium sedicola SBU-16 as a new endophytic taxol-producing fungus from Taxus baccata grown in Iran.

    PubMed

    Mirjalili, Mohammad Hossein; Farzaneh, Mohsen; Bonfill, Mercedes; Rezadoost, Hassan; Ghassempour, Alireza

    2012-03-01

    In this study, a total of 25 endophytic fungi were successfully isolated from the inner bark of Taxus baccata grown in Iran by the aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction (PCR) analysis for the presence of the Taxus taxadiene synthase (ts) gene, which encodes the enzyme catalyzing the first committed step of taxol biosynthesis. Four of 25 isolated endophytic fungi isolates showed PCR positive for the ts gene. Subsequently, taxol and 10-deacetylbaccatin III (10-DAB III) were extracted from culture filtrates and mycelia of the PCR positive isolates and analyzed by high-performance liquid chromatography and mass spectrometry. The analysis showed that one isolate (SBU-16) produced taxol (6.9 ± 0.2 μg L(-1) ) and its intermediate compound, 10-DAB III (2.2 ± 0.1 μg L(-1) ). The isolate SBU-16 was identified as Stemphylium sedicola SBU-16, according to its morphological characteristics as well as the internal transcribed spacer nuclear rDNA gene sequence analysis. Interestingly, this is the first report of the genus Stemphylium as a taxol-producing taxon.

  2. Exploitation of phage battery in the search for bioactive actinomycetes.

    PubMed

    Kurtböke, D Ipek

    2011-02-01

    Screening of microbial natural products continues to represent an important route to the discovery of novel bioactive compounds for the development of new therapeutic agents, and actinomycetes are still the major producers of biopharmaceuticals. Selective isolation of bioactive actinomycete species, in particular the rare ones, has thus become a target for industrial microbiologists. In this context, bacteriophages have proven to be useful tools as (1) naturally present indicators of under-represented or rare actinomycete taxa in environmental samples, (2) indicators of the relatedness of bioactive taxa in target-directed search and discovery, (3) de-selection agents of unwanted taxa on isolation plates in target-specific search for rare actinomycete taxa, (4) tools in screening assays for specific targets. Against this background, a number of case studies are presented to illustrate the use of bacteriophages as tools in actinomycete-origin bioactive compound search and discovery programs.

  3. Molecular characterization and identification of plant growth promoting endophytic bacteria isolated from the root nodules of pea (Pisum sativum L.).

    PubMed

    Tariq, Mohsin; Hameed, Sohail; Yasmeen, Tahira; Zahid, Mehwish; Zafar, Marriam

    2014-02-01

    Root nodule accommodates various non-nodulating bacteria at varying densities. Present study was planned to identify and characterize the non-nodulating bacteria from the pea plant. Ten fast growing bacteria were isolated from the root nodules of cultivated pea plants. These bacterial isolates were unable to nodulate pea plants in nodulation assay, which indicate the non-rhizobial nature of these bacteria. Bacterial isolates were tested in vitro for plant growth promoting properties including indole acetic acid (IAA) production, nitrogen fixation, phosphate solubilization, root colonization and biofilm formation. Six isolates were able to produce IAA at varying level from 0.86 to 16.16 μg ml(-1), with the isolate MSP9 being most efficient. Only two isolates, MSP2 and MSP10, were able to fix nitrogen. All isolates were able to solubilize inorganic phosphorus ranging from 5.57 to 11.73 μg ml(-1), except MSP4. Bacterial isolates showed considerably better potential for colonization on pea roots. Isolates MSP9 and MSP10 were most efficient in biofilm formation on polyvinyl chloride, which indicated their potential to withstand various biotic and abiotic stresses, whereas the remaining isolates showed a very poor biofilm formation ability. The most efficient plant growth promoting agents, MSP9 and MSP10, were phylogenetically identified by 16S rRNA gene sequence analysis as Ochrobactrum and Enterobacter, respectively, with 99% similarity. It is suggested the potential endophytic bacterial strains, Ochrobactrum sp. MSP9 and Enterobacter sp. MSP10, can be used as biofertilizers for various legume and non-legume crops after studying their interaction with the host crop and field evaluation.

  4. Endophytic Fungal Flora from Roots and Fruits of an Indian Neem Plant Azadirachta indica A. Juss., and Impact of Culture Media on their Isolation.

    PubMed

    Verma, Vijay C; Gond, Surendra K; Kumar, Anuj; Kharwar, Ravindra N; Boulanger, Lori-Ann; Strobel, Gary A

    2011-10-01

    Azadirachta indica A. Juss. (neem), native to India, is well known worldwide for its insecticidal and ethanopharmacological properties. Although endophytic microbes are known from this plant as only leaves and stems were the subjects of past reports. Now, a variety of procedures and a number of different media were used to isolate the maximum number of endophytic fungi from unripe fruits and roots. A total of 272 isolates of 29 filamentous fungal taxa were isolated at rate of 68.0% from 400 samples of three different individual trees (at locations-Az1, Az2, Az3). Mycological agar (MCA) medium yielded the highest number of isolates (95, with a 14.50% isolation rate) with the greatest species richness. Mycelia Sterilia (1, 2, 3) accounted for 11.06%, Coelomycetes 7.25%, while Hyphomycetes showed the maximum number of representative isolates (81.69%). Mycelia-Sterilia (1, 2, 3), based on their 5.8S ITS 1, ITS2 and partial 18S and 28S rDNA sequences were identified as Fusarium solani (99%), Chaetomium globosum (93%) and Chaetomium globosum (93%) respectively. Humicola, Drechslera, Colletotrichum, and Scytalidium sp. were some of the peculiar fungal endophytes recovered from this plant.

  5. Indolediketopiperazine Alkaloids from Eurotium cristatum EN-220, an Endophytic Fungus Isolated from the Marine Alga Sargassum thunbergii.

    PubMed

    Du, Feng-Yu; Li, Xin; Li, Xiao-Ming; Zhu, Li-Wei; Wang, Bin-Gui

    2017-01-25

    Four new indolediketopiperazine derivatives (1-4), along with nine known congeners (5-13), were isolated and identified from the culture extract of Eurotium cristatum EN-220, an endophytic fungus obtained from the marine alga Sargassum thunbergii. The structures of thesecompounds were elucidated on the basis of extensive spectroscopic analysis and the absolute configurations of compounds 1-4 were established by NOESY experiments and by chiral HPLC analyses of their acid hydrolysates. The absolute configuration of C-8 (a quaternary carbon substituted with a hydroxyl group) in 5 of preechinulin class was firstly determined by electronic circular dichroism (ECD) calculations. All these compounds were evaluatedfor brine shrimp (Artemia salina) lethality and nematicidal activity as well as antioxidativeand antimicrobial potency.

  6. Indolediketopiperazine Alkaloids from Eurotium cristatum EN-220, an Endophytic Fungus Isolated from the Marine Alga Sargassum thunbergii

    PubMed Central

    Du, Feng-Yu; Li, Xin; Li, Xiao-Ming; Zhu, Li-Wei; Wang, Bin-Gui

    2017-01-01

    Four new indolediketopiperazine derivatives (1–4), along with nine known congeners (5–13), were isolated and identified from the culture extract of Eurotium cristatum EN-220, an endophytic fungus obtained from the marine alga Sargassum thunbergii. The structures of thesecompounds were elucidated on the basis of extensive spectroscopic analysis and the absolute configurations of compounds 1–4 were established by NOESY experiments and by chiral HPLC analyses of their acid hydrolysates. The absolute configuration of C-8 (a quaternary carbon substituted with a hydroxyl group) in 5 of preechinulin class was firstly determined by electronic circular dichroism (ECD) calculations. All these compounds were evaluatedfor brine shrimp (Artemia salina) lethality and nematicidal activity as well as antioxidativeand antimicrobial potency. PMID:28125012

  7. Anti-malarial Activities of Two Soil Actinomycete Isolates from Sabah via Inhibition of Glycogen Synthase Kinase 3β

    PubMed Central

    Dahari, Dhiana Efani; Salleh, Raifana Mohamad; Mahmud, Fauze; Chin, Lee Ping; Embi, Noor; Sidek, Hasidah Mohd

    2016-01-01

    Exploiting natural resources for bioactive compounds is an attractive drug discovery strategy in search for new anti-malarial drugs with novel modes of action. Initial screening efforts in our laboratory revealed two preparations of soil-derived actinomycetes (H11809 and FH025) with potent anti-malarial activities. Both crude extracts showed glycogen synthase kinase 3β (GSK3β)-inhibitory activities in a yeast-based kinase assay. We have previously shown that the GSK3 inhibitor, lithium chloride (LiCl), was able to suppress parasitaemia development in a rodent model of malarial infection. The present study aims to evaluate whether anti-malarial activities of H11809 and FH025 involve the inhibition of GSK3β. The acetone crude extracts of H11809 and FH025 each exerted strong inhibition on the growth of Plasmodium falciparum 3D7 in vitro with 50% inhibitory concentration (IC50) values of 0.57 ± 0.09 and 1.28 ± 0.11 µg/mL, respectively. The tested extracts exhibited Selectivity Index (SI) values exceeding 10 for the 3D7 strain. Both H11809 and FH025 showed dosage-dependent chemo-suppressive activities in vivo and improved animal survivability compared to non-treated infected mice. Western analysis revealed increased phosphorylation of serine (Ser 9) GSK3β (by 6.79 to 6.83-fold) in liver samples from infected mice treated with H11809 or FH025 compared to samples from non-infected or non-treated infected mice. A compound already identified in H11809 (data not shown), dibutyl phthalate (DBP) showed active anti-plasmodial activity against 3D7 (IC50 4.87 ± 1.26 µg/mL which is equivalent to 17.50 µM) and good chemo-suppressive activity in vivo (60.80% chemo-suppression at 300 mg/kg body weight [bw] dosage). DBP administration also resulted in increased phosphorylation of Ser 9 GSK3β compared to controls. Findings from the present study demonstrate that the potent anti-malarial activities of H11809 and FH025 were mediated via inhibition of host GSK3β. In addition

  8. Marine actinomycetes: an ongoing source of novel bioactive metabolites.

    PubMed

    Subramani, Ramesh; Aalbersberg, William

    2012-12-20

    Actinomycetes are virtually unlimited sources of novel compounds with many therapeutic applications and hold a prominent position due to their diversity and proven ability to produce novel bioactive compounds. There are more than 22,000 known microbial secondary metabolites, 70% of which are produced by actinomycetes, 20% from fungi, 7% from Bacillus spp. and 1-2% by other bacteria. Among the actinomycetes, streptomycetes group are considered economically important because out of the approximately more than 10,000 known antibiotics, 50-55% are produced by this genus. The ecological role of actinomycetes in the marine ecosystem is largely neglected and various assumptions meant there was little incentive to isolate marine strains for search and discovery of new drugs. The search for and discovery of rare and new actinomycetes is of significant interest to drug discovery due to a growing need for the development of new and potent therapeutic agents. Modern molecular technologies are adding strength to the target-directed search for detection and isolation of bioactive actinomycetes, and continued development of improved cultivation methods and molecular technologies for accessing the marine environment promises to provide access to this significant new source of chemical diversity with novel/rare actinomycetes including new species of previously reported actinomycetes. Copyright © 2012 Elsevier GmbH. All rights reserved.

  9. Negative effects on survival and performance of Norway spruce seedlings colonized by dark septate root endophytes are primarily isolate-dependent.

    PubMed

    Tellenbach, Christoph; Grünig, Christoph R; Sieber, Thomas N

    2011-09-01

    Root endophytes are common and genetically highly diverse suggesting important ecological roles. Yet, relative to above-ground endophytes, little is known about them. Dark septate endophytic fungi of the Phialocephala fortinii s.l.-Acephala applanata species complex (PAC) are ubiquitous root colonizers of conifers and Ericaceae, but their ecological function is largely unknown. Responses of Norway spruce seedlings of two seed provenances to inoculations with isolates of four PAC species were studied in vitro. In addition, isolates of Phialocephala subalpina from two populations within and one outside the natural range of Norway spruce were also included to study the effect of the geographic origin of P. subalpina on host response. The interaction of PAC with Norway spruce ranged from neutral to highly virulent and was primarily isolate-dependent. Variation in virulence was much higher within than among species, nonetheless only isolates of P. subalpina were highly virulent. Disease caused by P. subalpina genotypes from the native range of Norway spruce was more severe than that induced by genotypes from outside the natural distribution of Norway spruce. Virulence was not correlated with the phylogenetic relatedness of the isolates but was positively correlated with the extent of fungal colonization as measured by quantitative real-time PCR.

  10. Endophytic Ability of Different Isolates of Entomopathogenic Fungi Beauveria bassiana (Balsamo) Vuillemin in Stem and Leaf Tissues of Maize (Zea mays L.).

    PubMed

    Renuka, S; Ramanujam, B; Poornesha, B

    2016-06-01

    The present study was conducted to examine the ability of six promising indigenous isolates of Beauveria bassiana (NBAII-Bb-5a, 7, 14, 19, 23 and 45) as an endophyte in maize stem and leaf tissues. Maize seedlings (var. Nithyashree) were inoculated with conidial suspensions and were examined for endophytic establishment in leaf and stems at different intervals during 15-90 days after treatment. All six isolates showed colonization in stem and leaf tissues with varying abilities of colonization and persistence. The mean percent colonization ranged from 7.41 to 20.37 % in older stem tissues and 3.70 to 21.29 % in young stem tissues and in leaf, it ranged from 6.46 to 27.78 % in older leaf tissues and 11.11 to 26.85 % in young leaf tissues. Among six isolates tested, Bb-23 isolate recorded the maximum mean colonization in older stem (20.37 %), older leaf (27.78 %) and in young stem (21.29 %). Bb-5a isolate showed maximum mean colonization in young leaf tissues (26.85 %). Persistence of inoculated fungal isolates decreased with increase in age of the plant. No physical symptoms of damage were observed in any of the B. bassiana treated plants. No colonization of B. bassiana was observed in the untreated control maize plants. The results obtained in plating and PCR techniques were similar with regard to the confirmation of endophytic establishment of B. bassiana. This study indicated the possibility of using B. bassiana as an endophyte in maize for management of maize stem borer, Chilo partellus.

  11. Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov., two novel actinomycetes isolated from muddy soil and emended description of the genus Actinomadura.

    PubMed

    Zhao, Junwei; Guo, Lifeng; Sun, Pengyu; Han, Chuanyu; Bai, Lu; Liu, Chongxi; Li, Yunxi; Xiang, Wensheng; Wang, Xiangjing

    2015-12-01

    Two novel actinomycetes, designated strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T), were isolated from muddy soil collected from a riverbank in Jiaohe, Jilin Province, north China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The 16S rRNA gene sequence analysis showed that the two novel isolates exhibited 99.9 % 16S rRNA gene sequence similarity with each other and that they are closely related to Actinomadura viridis IFO 15238(T) (99.6, 99.6 %) and Actinomadura vinacea IFO 14688(T) (99.3, 99.3 %). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the two cultures clustered together and formed a cluster with A. viridis IFO 15238(T), A. vinacea IFO 14688(T) and Actinomadura rugatobispora IFO 14382(T). However, the DNA-DNA hybridization value between strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) was 63.6 %, and the values between the two strains and their close phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their close phylogenetic relatives. Thus, strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) represent two novel species of the genus Actinomadura, for which the names Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov. are proposed. The type strains are NEAU-Jh1-3(T) (=CGMCC 4.7197(T) = JCM 30341(T)) and NEAU-Jh2-5(T) (=CGMCC 4.7211(T) = JCM 30342(T)), respectively.

  12. Purification and biochemical characterization of a detergent-stable keratinase from a newly thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 isolated from poultry compost.

    PubMed

    Habbeche, Amina; Saoudi, Boudjema; Jaouadi, Bassem; Haberra, Soumaya; Kerouaz, Bilal; Boudelaa, Mokhtar; Badis, Abdelmalek; Ladjama, Ali

    2014-04-01

    An extracellular thermostable keratinase (KERAK-29) was purified and biochemically characterized from a thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 newly isolated from Algerian poultry compost. The isolate exhibited high keratinase production when grown in chicken feather meal media (24,000 U/ml). Based on matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis, the purified enzyme is a monomer with a molecular mass of 29,233.10-Da. The data revealed that the 25 N-terminal residue sequence displayed by KERAK-29 was TQADPPSWGLNNIDRQTAFTKATSI, which showed high homology with those of Streptomyces proteases. This keratinase was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP), which suggests that it belongs to the serine protease family. Using keratin azure as a substrate, the optimum pH and temperature values for keratinase activity were pH 10 and 70°C, respectively. KERAK-29 was stable between 20 and 60°C and pH 3 and 10 for 5 and 120 h, respectively, and its thermoactivity and thermostability were enhanced in the presence of 5 mM Mn(2+). Its catalytic efficiency was higher than that of the KERAB keratinase from Streptomyces sp. strain AB1. KERAK-29 was also noted to show high keratinolytic activity and significant stability in the presence of detergents, which made it able to accomplish the entire feather-biodegradation process on its own. The ability of the A. keratinilytica strain Cpt29 to grow and produce substantial levels of keratinase using feather as a substrate could open new promising opportunities for the valorization of keratin-containing wastes and reduction of its impacts on the environment.

  13. Isolation and Characterization of “Terrein” an Antimicrobial and Antitumor Compound from Endophytic Fungus Aspergillus terreus (JAS-2) Associated from Achyranthus aspera Varanasi, India

    PubMed Central

    Goutam, Jyoti; Sharma, Gunjan; Tiwari, Vinod K.; Mishra, Amrita; Kharwar, Ravindra N.; Ramaraj, Vijayakumar; Koch, Biplob

    2017-01-01

    The present study aimed at characterizing biological potentials of endophyte Aspergillus terreus JAS-2 isolated from Achyranthus aspera. Crude extracted from endophytic fungus JAS-2 was purified and chemically characterized by chromatographic and spectroscopic studies respectively. Spectral assignment of NMR (nuclear magnetic resonance) data, 1H proton and 13C carbon analysis along with FTIR data elucidated the structure of compound as 4,5-Dihydroxy-3-(1-propenyl)-2-cyclopenten-1-one. After purification and identification a set of experiment was conducted to explore efficacy of compound. Results revealed that on accessing the antifungal activity of compound, growth diameter of tested phytopathogenic fungi was reduced to 50% at higher concentration taken (10 μgμl−1). Compound exhibited in-vitro bacterial cell inhibition at 20 μgml−1 concentration along with moderate antioxidant behavior. Evaluation of anticancer activity against human lung cancer cell line (A-549) exhibited its IC50 value to be 121.9 ± 4.821 μgml−1. Further cell cycle phase distribution were analyzed on the basis of DNA content and evaluated by FACS (Fluorescence Activated Cell Sorting) and it was revealed that at 150 μgml−1 of compound maximum cells were found in sub G1 phase which represents apoptotic dead cells. Terrein (4, 5-Dihydroxy-3-(1-propenyl)-2-cyclopenten-1-one) a multi-potential was isolated from endophytic fungus JAS-2, from well recognized medicinal herb A. aspera. To best of our knowledge, this is the first report of “Terrein” from endophytic derived fungus. This compound had also exhibited anticancer and antifungal activity against human lung cancer cell line A-549 and Bipolaris sorokiniana respectively which is causal organism of many plants disease. Hence endophytes are serving as alternative sources of drug molecules. PMID:28790982

  14. Bioactive endophytic fungi isolated from Caesalpinia echinata Lam. (Brazilwood) and identification of beauvericin as a trypanocidal metabolite from Fusarium sp.

    PubMed Central

    Campos, Fernanda Fraga; Sales, Policarpo A; Romanha, Alvaro José; Araújo, Márcio SS; Siqueira, Ezequias P; Resende, Jarbas M; Alves, Tânia MA; Martins-Filho, Olindo A; dos Santos, Vera Lúcia; Rosa, Carlos A; Zani, Carlos L; Cota, Betania Barros

    2015-01-01

    Aiming to identify new sources of bioactive secondary metabolites, we isolated 82 endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro assays. The organic extracts from three isolates showed antibacterial activity against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration (MIC) 32-64 μg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC 64 μg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 μg/mL), Candida albicans and Candida tropicalis (MIC 64-128 μg/mL). Fourteen extracts at a concentration of 20 μg/mL showed antitumour activities against human breast cancer and human renal cancer cells, while two isolates showed anti-tumour activities against human melanoma cancer cells. Six extracts were able to reduce the proliferation of human peripheral blood mononuclear cells, indicating some degree of selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania) amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 μg/mL. The trypanocidal extract obtained from Fusarium sp. [KF611679] culture was subjected to bioguided fractionation, which revealed beauvericin as the compound responsible for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a half maximal inhibitory concentration of 1.9 μg/mL (2.43 μM) in a T. cruzi cellular culture assay. PMID:25742265

  15. Analysis and characterization of cultivable heavy metal-resistant bacterial endophytes isolated from Cd-hyperaccumulator Solanum nigrum L. and their potential use for phytoremediation.

    PubMed

    Luo, Sheng-lian; Chen, Liang; Chen, Jue-liang; Xiao, Xiao; Xu, Tao-ying; Wan, Yong; Rao, Chan; Liu, Cheng-bin; Liu, Yu-tang; Lai, Cui; Zeng, Guang-ming

    2011-11-01

    This study investigates the heavy metal-resistant bacterial endophytes of Cd-hyperaccumulator Solanum nigrum L. grown on a mine tailing by using cultivation-dependent technique. Thirty Cd-tolerant bacterial endophytes were isolated from roots, stems, and leaves of S. nigrum L. and classified by amplified ribosomal DNA-restriction analysis into 18 different types. Phylogenetic analysis based on 16S rDNA sequences showed that these isolates belonged to four groups: Actinobacteria (43%), Proteobacteria (23%), Bacteroidetes (27%) and Firmicutes (7%). All the isolates were then characterized for their plant growth promoting traits as well as their resistances to different heavy metals; and the actual plant growth promotion and colonization ability were also assessed. Four isolates were re-introduced into S. nigrum L. under Cd stress and resulted in Cd phytotoxicity decrease, as dry weights of roots increased from 55% to 143% and dry weights of above-ground from 64% to 100% compared to the uninoculated ones. The total Cd accumulation of inoculated plants increased from 66% to 135% (roots) and from 22% to 64% (above-ground) compared to the uninoculated ones. Our research suggests that bacterial endophytes are a most promising resource and may be the excellent candidates of bio-inoculants for enhancing the phytoremediation efficiency.

  16. Isolation of a rice endophytic bacterium, Pantoea sp. Sd-1, with ligninolytic activity and characterization of its rice straw degradation ability.

    PubMed

    Xiong, X Q; Liao, H D; Ma, J S; Liu, X M; Zhang, L Y; Shi, X W; Yang, X L; Lu, X N; Zhu, Y H

    2014-02-01

    This study focused on an endophytic bacterial strain, Pantoea sp. Sd-1, which can be used to degrade lignin and rice straw. This strain was isolated from rice seeds by an optimized surface sterilization method. Pantoea sp. Sd-1 showed exceptional ability to degrade rice straw and lignin. In rice straw or kraft lignin-containing medium supplemented with 1% glucose and 0.5% peptone, Pantoea sp. Sd-1 effectively reduced the rice straw mass weight by 54.5% after 6 days of treatment. The strain was also capable of reducing the lignin colour (52.4%) and content (69.1%) after 4 days of incubation. The findings suggested that the rice endophytic bacterium Pantoea sp. Sd-1 could be applied for the degradation of lignocellulose biomass, such as rice straw. Rice straw, an abundant agricultural by-product in China, is very difficult to degrade because of its high lignin content. Due to the immense environmental adaptability and biochemical versatility of bacteria, endophytic bacteria are useful resources for biodegradation. In this study, we screened for endophytic bacteria capable of biodegrading rice straw and lignin and obtained one strain, Pantoea sp. Sd-1, with suitable characteristics. Sd-1 could be used for degradation of rice straw and lignin, and may play an important role in biodegradation of this agricultural by-product. © 2013 The Society for Applied Microbiology.

  17. Colonization and plant growth promoting characterization of endophytic Pseudomonas chlororaphis strain Zong1 isolated from Sophora alopecuroides root nodules

    PubMed Central

    Zhao, Long Fei; Xu, Ya Jun; Ma, Zhan Qiang; Deng, Zhen Shan; Shan, Chang Juan; Wei, Ge Hong

    2013-01-01

    The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC) and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4–60 ºC for 15 min, had a wide range pH tolerance of 6.0–11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (μg/mL): Phosphonomycin (100), Penicillin (100) and Ampicillin (100). It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v) methylene blue and 0.1–0.2% (w/v) methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp.) showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p < 0.05) when compared to a negative control. Therefore, as an endophyte P. chlororaphis Zong1 may play important roles as a potential plant-growth promoting agent. PMID:24294262

  18. Colonization and plant growth promoting characterization of endophytic Pseudomonas chlororaphis strain Zong1 isolated from Sophora alopecuroides root nodules.

    PubMed

    Zhao, Long Fei; Xu, Ya Jun; Ma, Zhan Qiang; Deng, Zhen Shan; Shan, Chang Juan; Wei, Ge Hong

    2013-01-01

    The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC) and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4-60 ºC for 15 min, had a wide range pH tolerance of 6.0-11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (μg/mL): Phosphonomycin (100), Penicillin (100) and Ampicillin (100). It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v) methylene blue and 0.1-0.2% (w/v) methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp.) showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p < 0.05) when compared to a negative control. Therefore, as an endophyte P. chlororaphis Zong1 may play important roles as a potential plant-growth promoting agent.

  19. Draft Genome Sequence of the Endophytic Strain Rhodococcus kyotonensis KB10, a Potential Biodegrading and Antibacterial Bacterium Isolated from Arabidopsis thaliana

    PubMed Central

    Hong, Chi Eun; Jo, Sung Hee

    2016-01-01

    Rhodococcus kyotonensis KB10 is an endophytic bacterium isolated from Arabidopsis thaliana. The organism showed mild antibacterial activity against the phytopathogen Pseudomonas syringae pv. tomato DC3000. This study reports the genome sequence of R. kyotonensis KB10. This bacterium contains an ectoine biosynthesis gene cluster and has the potential to degrade nitroaromatic compounds. The identified bacterium may be a suitable biocontrol agent and degrader of environmental pollutants. PMID:27389269

  20. Draft Genome Sequence of Curtobacterium sp. Strain ER1/6, an Endophytic Strain Isolated from Citrus sinensis with Potential To Be Used as a Biocontrol Agent

    PubMed Central

    Garrido, Leandro Maza; Alves, João Marcelo Pereira; Oliveira, Liliane Santana; Padilla, Gabriel

    2016-01-01

    Herein, we report a draft genome sequence of the endophytic Curtobacterium sp. strain ER1/6, isolated from a surface-sterilized Citrus sinensis branch, and it presented the capability to control phytopathogens. Functional annotation of the ~3.4-Mb genome revealed 3,100 protein-coding genes, with many products related to known ecological and biotechnological aspects of this bacterium. PMID:27856581

  1. Complete genome sequence of Streptomyces sp. strain CFMR 7, a natural rubber degrading actinomycete isolated from Penang, Malaysia.

    PubMed

    Nanthini, Jayaram; Chia, Kim-Hou; Thottathil, Gincy P; Taylor, Todd D; Kondo, Shinji; Najimudin, Nazalan; Baybayan, Primo; Singh, Siddharth; Sudesh, Kumar

    2015-11-20

    Streptomyces sp. strain CFMR 7, which naturally degrades rubber, was isolated from a rubber plantation. Whole genome sequencing and assembly resulted in 2 contigs with total genome size of 8.248 Mb. Two latex clearing protein (lcp) genes which are responsible for rubber degrading activities were identified.

  2. Flindersiella endophytica gen. nov., sp. nov., an endophytic actinobacterium isolated from the root of Grey Box, an endemic eucalyptus tree.

    PubMed

    Kaewkla, Onuma; Franco, Christopher M M

    2011-09-01

    A novel endophytic actinobacterium, designated strain EUM 378(T), was isolated from the surface-sterilized root tissue of Eucalyptus microcarpa, a eucalyptus tree known as Grey Box. Phylogenetic evaluation based on 16S rRNA gene sequence analysis, including alignment with taxon-specific 16S rRNA gene signature nucleotides, placed this isolate as a member of the family Nocardioidaceae. Strain EUM 378(T) showed >5.5 % 16S rRNA gene sequence divergence from other members of this family and was related most closely to Actinopolymorpha alba YIM 48868(T) (94.2 %) and Actinopolymorpha singaporensis IM 7744(T) (94.4 %). This Gram-positive, aerobic actinobacterium has well-developed substrate mycelia that fragment into small rods. Chemotaxonomic data revealed that the cell wall contains LL-diaminopimelic acid, ribose, glucose and rhamnose. MK-10(H₆) is the predominant menaquinone. Chemotaxonomic and phylogenetic evidence confirmed that strain EUM 378(T) represents a novel species of a new genus, for which the name Flindersiella endophytica gen. nov., sp. nov. is proposed. The type strain is EUM 378(T) ( = DSM 45355(T) = ACM 5289(T)).

  3. Biotransformation of 2-Benzoxazolinone and 2-Hydroxy-1,4-Benzoxazin-3-one by Endophytic Fungi Isolated from Aphelandra tetragona

    PubMed Central

    Zikmundová, M.; Drandarov, K.; Bigler, L.; Hesse, M.; Werner, C.

    2002-01-01

    The biotransformation of the phytoanticipins 2-benzoxazolinone (BOA) and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) by four endophytic fungi isolated from Aphelandra tetragona was studied. Using high-performance liquid chromatography-mass spectrometry, several new products of acylation, oxidation, reduction, hydrolysis, and nitration were identified. Fusarium sambucinum detoxified BOA and HBOA to N-(2-hydroxyphenyl)malonamic acid. Plectosporium tabacinum, Gliocladium cibotii, and Chaetosphaeria sp. transformed HBOA to 2-hydroxy-N-(2-hydroxyphenyl)acetamide, N-(2-hydroxyphenyl)acetamide, N-(2-hydroxy-5-nitrophenyl)acetamide, N-(2-hydroxy-3-nitrophenyl)acetamide, 2-amino-3H-phenoxazin-3-one, 2-acetylamino-3H-phenoxazin-3-one, and 2-(N-hydroxy)acetylamino-3H-phenoxazin-3-one. BOA was not degraded by these three fungal isolates. Using 2-hydroxy-N-(2-hydroxyphenyl)[13C2]acetamide, it was shown that the metabolic pathway for HBOA and BOA degradation leads to o-aminophenol as a key intermediate. PMID:12324332

  4. Biotransformation of 2-benzoxazolinone and 2-hydroxy-1,4-benzoxazin-3-one by endophytic fungi isolated from Aphelandra tetragona.

    PubMed

    Zikmundová, M; Drandarov, K; Bigler, L; Hesse, M; Werner, C

    2002-10-01

    The biotransformation of the phytoanticipins 2-benzoxazolinone (BOA) and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) by four endophytic fungi isolated from Aphelandra tetragona was studied. Using high-performance liquid chromatography-mass spectrometry, several new products of acylation, oxidation, reduction, hydrolysis, and nitration were identified. Fusarium sambucinum detoxified BOA and HBOA to N-(2-hydroxyphenyl)malonamic acid. Plectosporium tabacinum, Gliocladium cibotii, and Chaetosphaeria sp. transformed HBOA to 2-hydroxy-N-(2-hydroxyphenyl)acetamide, N-(2-hydroxyphenyl)acetamide, N-(2-hydroxy-5-nitrophenyl)acetamide, N-(2-hydroxy-3-nitrophenyl)acetamide, 2-amino-3H-phenoxazin-3-one, 2-acetylamino-3H-phenoxazin-3-one, and 2-(N-hydroxy)acetylamino-3H-phenoxazin-3-one. BOA was not degraded by these three fungal isolates. Using 2-hydroxy-N-(2-hydroxyphenyl)[(13)C(2)]acetamide, it was shown that the metabolic pathway for HBOA and BOA degradation leads to o-aminophenol as a key intermediate.

  5. Trienylfuranol A and trienylfuranone A-B: metabolites isolated from an endophytic fungus, Hypoxylon submoniticulosum, in the raspberry Rubus idaeus.

    PubMed

    Burgess, Kevin M N; Ibrahim, Ashraf; Sørensen, Dan; Sumarah, Mark W

    2017-03-01

    A strain of Hypoxylon submonticulosum was isolated as an endophyte from a surface-sterilized leaf of a cultivated raspberry (Rubus idaeus). The liquid culture extract displayed growth inhibition activity against Saccharomyces cerevisiae using a disc diffusion assay. The extract's major component was identified as a new natural product, trienylfuranol A (1S,2S,4R)-1-((1'E,3'E)-hexa-1',3',5'-trienyl)-tetrahydro-4-methylfuran-2-ol (1), by high-resolution LC-MS and 1D and 2D NMR spectroscopy. Two additional new metabolites, trienylfuranones A (2) and B (3), were isolated as minor components of the extract and their structure elucidation revealed that they were biosynthetically related to 1. Absolute stereochemical configurations of compounds 1-3 were confirmed by NOE NMR experiments and by the preparation of Mosher esters. Complete hydrogenation of 1 yielded tetrahydrofuran 7 that was used for stereochemical characterization and assessment of antifungal activity.The Journal of Antibiotics advance online publication, 1 March 2017; doi:10.1038/ja.2017.18.

  6. Actinomycetes in Karstic caves of northern Spain (Altamira and Tito Bustillo).

    PubMed

    Groth, I; Vettermann, R; Schuetze, B; Schumann, P; Saiz-Jimenez, C

    1999-05-01

    A variety of isolation procedures were carried out to study the involvement of bacteria in the colonisation and biodeterioration of Spanish caves with paleolithic rock art (Altamira and Tito Bustillo). The applied techniques mainly aimed to isolate heterotrophic bacteria such as streptomycetes, nocardioform and coryneform actinomycetes, and other gram-positive and gram-negative bacteria. The results demonstrated that actinomycetes were the most abundant gram-positive bacteria in the caves. Actinomycetes revealed a great taxonomic diversity with the predominant isolates belonging to the genus Streptomyces. Members of the genera Nocardia, Rhodococcus, Nocardioides, Amycolatopsis, Saccharothrix, Brevibacterium, Microbacterium, and coccoid actinomycetes (family Micrococcaceae) were also found.

  7. Antibacterial activity of some actinomycetes from Tamil Nadu, India

    PubMed Central

    Kumar, Pachaiyappan Saravana; Raj, John Poonga Preetam; Duraipandiyan, Veeramuthu; Ignacimuthu, Savarimuthu

    2012-01-01

    Objective To isolate novel actinomycetes and to evaluate their antibacterial activity. Methods Three soil samples were collected from Vengodu (village) in Kanchipuram district, Tamil Nadu, India. Actinomycetes were isolated using serial dilution and plating method on actinomycetes isolation agar. Results Totally 35 isolates were obtained on the basis of colony characteristics on actinomycetes isolation agar. All the isolates were screened for antibacterial activity by cross streak method. Medium and optimization of day were done for the potent strains using Nathan's agar well diffusion method. Isolation of bioactive compounds from significant active isolates was done by using different media. The most active isolate VAS 10 was identified as Actinobacterium Loyola PBT VAS 10 (accession No. JF501398) using 16s rRNA sequence method. The hexane, ethyl acetate, dichloromethane and butanol extracts of VAS 10 were tested against bacteria. The maximum antibacterial activity was observed in dichloromethane and ethyl acetate; maximum zones of inhibition were observed against Enterococcus durans. The rRNA secondary structure and the restriction sites of Actinobacterium Loyola VAS 10 were predicted using Genebee and NEBCutter online tools respectively. Conclusions The present study showed that among the isolated actinomycetes, Actinobacterium Loyola PBT VAS 10 (accession No. JF501398) showed good antibacterial activity against the tested bacteria. PMID:23593572

  8. Production and characterization of a thermostable alpha-amylase from Nocardiopsis sp. endophyte of yam bean.

    PubMed

    Stamford, T L; Stamford, N P; Coelho, L C; Araújo, J M

    2001-01-01

    Thermostable amylolytic enzymes have been currently investigated to improve industrial processes of starch degradation. Studies on production of alpha-amylase by Nocardiopsis sp., an endophytic actinomycete isolated from yam bean (Pachyrhizus erosus L. Urban), showed that higher enzyme levels were obtained at the end of the logarithmic growth phase after incubation for 72 h at pH 8.6. Maximum activity of alpha-amylase was obtained at pH 5.0 and 70 degrees C. The isolated enzyme exhibited thermostable properties as indicated by retention of 100% of residual activity at 70 degrees C, and 50% of residual activity at 90 degrees C for 10 min. Extracellular enzyme from Nocardiopsis sp. was purified by fractional precipitation with ammonium sulphate. After 60% saturation produced 1130 U mg-1 protein and yield was 28% with purification 2.7-fold. The enzyme produced by Nocardiopsis sp. has potential for industrial applications.

  9. Cultivation and detection of endophytic aerobic methanotrophs isolated from Sphagnum species as a perspective for environmental biotechnology

    PubMed Central

    2014-01-01

    Enriched cultures of microorganisms are an essential step in the production of inoculum of these organisms for biotechnology and bioengineering. The potential application of methanotrophic microorganisms for removal of methane produced from landfills and coal mines as well as biodegradation of toxic compounds has been widely studied. Therefore, searching for new sources of methanotrophs can contribute to increasing the possibilities of biotechnology and bioengineering. Enrichment cultures of endophytic methanotrophs from Sphagnum sp. were initiated in NMS medium, a most widely used medium for cultivation of methanotrophic bacteria from various environments proposed in 1970 by Whittenbury. Incubation was carried out at 10, 20, 30, and 37°C with vigorous shaking on a shaker (180 rpm). The source of carbon and energy for endophytes were methane at the concentration range between 1-20%. It appeared that the consortium of endophytic bacteria grew only at the temperature of 20 and 30°C. During the culture of endophytes, the measurements of gas concentration showed a steady loss of methane and oxygen, as well as accumulation of carbon dioxide as a CH4 oxidation product. The use of FISH has made characterization of endophytic consortia possible. It turned out that the population of endophytes consists of type I and II methanotrophs as well as associated non-methanotrophic bacteria. Furthermore, we determined the potential of the examined bacteria for methane oxidation, which ranged up to 4,7 μMCH4 per ml of the population of endophytes per day. PMID:25401064

  10. Aspergillus clavatus Y2H0002 as a New Endophytic Fungal Strain Producing Gibberellins Isolated from Nymphoides pe ltata in Fresh Water.

    PubMed

    You, Young-Hyun; Kwak, Tae Won; Kang, Sang-Mo; Lee, Myung-Chul; Kim, Jong-Guk

    2015-03-01

    Eighteen endophytic fungi with different colony morphologies were isolated from the roots of Nymphoides peltata growing in the Dalsung wetland. The fungal culture filtrates of the endophytic fungi were treated to Waito-c rice seedling to evaluate their plant growth-promoting activities. Culture filtrate of Y2H0002 fungal strain promoted the growth of the Waito-c rice seedlings. This strain was identified on the basis of sequences of the partial internal transcribed spacer region and the partial beta-tubulin gene. Upon chromatographic analysis of the culture filtrate of Y2H0002 strain, the gibberellins (GAs: GA1, GA3, and GA4) were detected and quantified. Molecular and morphological studies identified the Y2H0002 strain as belonging to Aspergillus clavatus. These results indicated that A. clavatus improves the growth of plants and produces various GAs, and may participate in the growth of plants under diverse environmental conditions.

  11. Isolation and Identification of the Antimicrobial Agent Beauvericin from the Endophytic Fusarium oxysporum 5-19 with NMR and ESI-MS/MS

    PubMed Central

    Ruan, Chuanfen; Bai, Xuelian; Zhang, Miao; Zhu, Shuangshuang; Jiang, Yingying

    2016-01-01

    Endophytic microbe has been proved to be one of rich sources of bioactive natural products with potential application for new drug and pesticide discovery. One cyclodepsipeptide, beauvericin, was firstly isolated from the fermentation broth of Fusarium oxysporum 5-19 endophytic on Edgeworthia chrysantha Linn. Its chemical structure was unambiguously identified by a combination of spectroscopic methods, such as HRESI-MS and 1H and 13C NMR. ESI-MS/MS was successfully used to elucidate the splitting decomposition route of the positive molecule ion of beauvericin. Antimicrobial results showed that this cyclodepsipeptide had inhibitory effect on three human pathogenic microbes, Candida albicans, Escherichia coli, and Staphylococcus aureus. In particular, beauvericin exhibited the strongest antimicrobial activity against S. aureus with MIC values of 3.91 μM, which had similar effect with that of the positive control amoxicillin. PMID:27413733

  12. Rhodococcus pedocola sp. nov. and Rhodococcus humicola sp. nov., two antibiotic-producing actinomycetes isolated from soil.

    PubMed

    Nguyen, Tuan Manh; Kim, Jaisoo

    2016-06-01

    Two novel actinobacterial strains, UC12T and UC33T, were isolated from forest topsoil in Suwon, Gyeonggi-Do, South Korea. Comparative analysis of nearly full-length 16S rRNA gene sequences of UC12T and UC33T revealed close pairwise similarity with species of the genus Rhodococcus, and the UC12T and UC33T sequences were most closely related to Rhodococcus canchipurensis MBRL 353T (98.91 % 16S rRNA gene sequence similarity) and Rhodococcus triatomae IMMIB RIV-085T (97.71 %), respectively. DNA-DNA hybridization showed 33.05-35.60 % genomic similarity between strains UC12T and UC33T, while strain UC12T shared DNA-DNA relatedness values of 32.71-41.29 % with the closest species of the genus Rhodococcus and strain UC33T shared 29.12-37.91 % genomic relatedness with the closest species of the genus Rhodococcus. Both strains showed similar chemotaxonomic characteristics. The major fatty acids were C16 : 0, summed feature 3, C18 : 1ω9c and C18 : 0 10-methyl. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major quinone derived was MK-8(H2). The cell-wall peptidoglycan contained meso-diaminopimelic acid, and galactose, glucose, arabinose and ribose were detected in whole cells. Mycolic acids were detected. The DNA G+C content of strains UC12T and UC33T was 72.7 mol% and 68.8 mol%, respectively. Both strains produced antibiotic(s) that inhibited bacterial pathogens but not fungi. Based on the physiological, biochemical and genotypic features and the DNA-DNA hybridization between the isolates and type strains of closely related species, we propose that these bacteria be classified as novel species of the genus Rhodococcus with the names Rhodococcus pedocola sp. nov. (type strain UC12T=KACC 18499T=NBRC 111580T) and Rhodococcus humicola sp. nov. (type strain UC33T=KACC 18500T=NBRC 111581T).

  13. Alloactinosynnema iranicum sp. nov., a rare actinomycete isolated from a hypersaline wetland, and emended description of the genus Alloactinosynnema.

    PubMed

    Nikou, Mahdi Moshtaghi; Ramezani, Mohaddaseh; Amoozegar, Mohammad Ali; Fazeli, Seyed Abolhassan Shahzadeh; Schumann, Peter; Spröer, Cathrin; Sánchez-Porro, Cristina; Ventosa, Antonio

    2014-04-01

    A Gram-staining-positive actinobacterial strain, Chem10(T), was isolated from soil around Inche-Broun hypersaline wetland in the north of Iran. Strain Chem10(T) was strictly aerobic, and catalase- and oxidase-positive. The isolate grew with 0-3 % NaCl, at 20-40 °C and at pH 6.0-8.0. The optimum temperature and pH for growth were 30 °C and pH 7.0, respectively. The cell wall of strain Chem10(T) contained meso-diaminopimelic acid as diamino acid and galactose, ribose and arabinose as whole-cell sugars. The polar lipid pattern contained diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain Chem10(T) synthesized cellular fatty acids of the straight-chain saturated and mono-unsaturated, and iso- and anteiso-branched types C14 : 0, C16 : 0, iso-C16 : 1, anteiso-C17 : 0, iso-C16 : 0, iso-C14 : 0 and iso-C15 : 0, and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 70.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Chem10(T) belonged to the family Pseudonocardiaceae and showed the closest phylogenetic similarity to Alloactinosynnema album KCTC 19294(T) (98.3 %) and Actinokineospora cibodasensis DSM 45658(T) (97.9 %). DNA-DNA relatedness values between the novel strain and strains Alloactinosynnema album KCTC 19294(T) and Actinokineospora cibodasensis DSM 45658(T) were only 52 % and 23 %, respectively. On the basis of phylogenetic analysis, phenotypic characteristics and DNA-DNA hybridization data, a novel species of the genus Alloactinosynnema is proposed, Alloactinosynnema iranicum sp. nov. The type strain is Chem10(T) ( = IBRC-M 10403(T) = CECT 8209(T)). In addition, an emended description of the genus Alloactinosynnema is proposed.

  14. Antiproliferative effect of extract from endophytic fungus Curvularia trifolii isolated from the "Veracruz Reef System" in Mexico.

    PubMed

    Couttolenc, Alan; Espinoza, Cesar; Fernández, José J; Norte, Manuel; Plata, Gabriela B; Padrón, José M; Shnyreva, Alla; Trigos, Ángel

    2016-08-01

    It is well known that marine fungi are an excellent source of biologically active secondary metabolites, and by 2011, it was reported that over 400 bioactive metabolites were derived from marine fungi. This study establishes the basis for future research on antiproliferative compounds of marine endophytes inhabited in the Veracruz Reef System. Isolation of the 34 fungal strains was carried out by microbiological method from samples of sponges, corals, and other biological material from the Veracruz Reef System. The fungal biomass and broth were separated and extracted with a mixture of solvents MeOH:CHCl3. Characterization and molecular identification of the fungal strains were performed through microbiological methods and the analysis of the ITS-rDNA regions. Antiproliferative activity was tested at a dose of 250 μg/mL on human solid tumor cell lines HBL-100, HeLa, SW1573, T-47D, and WiDr by the SRB assay after 48 h-exposure to the fungal extracts. The extracts from five isolates showed an antiproliferative effect against one or more of the tested cell lines (percentage growth < 50%). The mycelial extract from the isolate LAEE 03 manifested the highest activity against the five cell lines (% PG of 17 HBL-100, 19 HeLa, 23 SW1573, -6 T-47D, and 10 WiDr) and the strain was identified as Curvularia trifolii (Kauffman) Boedijn (Pleosporaceae). The results obtained indicate that the extract from a marine derived C. trifolii has the antiproliferative effect, thus suggesting that this organism is a good candidate for further analysis of its metabolites.

  15. A novel poly (L-lactide) degrading actinomycetes isolated from Thai forest soil, phylogenic relationship and the enzyme characterization.

    PubMed

    Sukkhum, Sukhumaporn; Tokuyama, Shinji; Tamura, Tomohiko; Kitpreechavanich, Vichien

    2009-12-01

    Thirteen poly (L-lactide)-degrading microorganisms were isolated and selected based on t