Endophytic Actinomycetes Research Articles

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

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 �/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.

Saccharopolyspora dendranthemae sp. nov. a halotolerant endophytic actinomycete isolated from a coastal salt marsh plant in Jiangsu, China

A halotolerant actinomycete strain, designated strain KLBMP 1305(T), was isolated from a salt marsh plant Dendranthema indicum (Linn.) Des Moul collected from the coastal region of Nantong, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP 1305(T) is a member of the genus Saccharopolyspora. The 16S rRNA gene sequence similarity indicated that strain KLBMP 1305(T) was most closely related to 'Saccharopolyspora pathumthaniensis' S582(T) (99.31 %), 'Saccharopolyspora endophytica' YIM 61095(T) (99.17 %) and Saccharopolyspora tripterygii YIM 65359(T) (99.15 %); similarity to other type strains of the genus Saccharopolyspora was <97.2 %. The organism had chemical and morphological features consistent with its classification in the genus Saccharopolyspora such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was MK-9(H4). The polar lipids detected were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unknown glycolipid and an unknown lipid. The major fatty acids were iso-C16:0, iso-C15:0, anteiso-C15:0, anteiso-C17:0 and sum in feature 8 (18:1ω7c/18:1ω6c). The G+C content of the genomic DNA of the type strain was 68.7 mol%. DNA-DNA relatedness data, together with phenotypic differences, clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate represents a novel species, for which the name Saccharopolyspora dendranthemae sp. nov. is proposed. The type strain is KLBMP 1305(T) (=KCTC 19889(T) = NBRC 108675(T)).

Isolation of endophytic streptomycetes from above- and belowground organs of Quercus serrata

The occurrence and localization of endophytic actinomycetes within diverse organs of host plants provide ecological information that can be used to evaluate the significance of their spatial habitats. The aim of this study was to isolate and characterize endophytic actinomycetes in different organs of Quercus serrata. For this purpose, actinomycete isolates were obtained from surface-sterilized tissues of both symptomless seedlings and aged trees of Q. serrata and rhizosphere soil of the sampled seedlings. Thirty-five isolates with the ability to sporulate, including 4 from leaves of the aged trees, 10, 6, and 15 from leaves, stems, and roots of the seedlings, respectively, and 8 soil-derived isolates, were selected and characterized. The 16S rDNA nucleotide sequence analyses revealed that all of them belonged to the genus Streptomyces. According to a neighbor-joining phylogenetic tree constructed based on the results, the isolates of plant origin were divided into three major clades with high bootstrap values of 98 or 99 %, whereas eight soil-derived isolates were located at different positions from those of the endophyte isolates. Moreover, two larger clades were formed, one of which contained isolates derived only from aboveground parts, while the other contained isolates from all of the organs. These results suggest that the endophytic streptomycetes in Q. serrata may differ in their habitat positions (i.e., either above- or belowground parts).

Screening of Antimicrobial and Antioxidant Secondary Metabolites from Endophytic Fungi Isolated from Wheat (Triticum Durum)

Abstract The emergence of antibiotic-resistant micro-organisms calls for inventive research and development strategies. Inhibition of these pathogenic micro-organisms may be a promising therapeutic approach. The screening of antimicrobial compounds from endophytes is a promising way to meet the increasing threat of drug-resistant strains of human and plant pathogens. In the present study, a total of 20 endophytic fungi and 23 endophytic actinomycetes have been isolated from wheat (Triticum durum). Mohamed Ben Bachir variety collected from Bordj Bou Arreridj region (Algeria) during winter 2010. The isolates were screened and evaluated for their antimicrobial and antioxidant activities. Antimicrobial activity was evaluated for crude ethyl acetate extracts using an agar diffusion assay against twelve pathogenic bacteria, yeast, and two phytopathogenic fungi. All extracts showed inhibitory activity on at least one or more pathogenic microorganisms, with an average zone of inhibition varied between 7 mm to 25 mm, and the largest zone was of 25 and 25.3 mm against candida albicans and Escherichia coli respectively. The antioxidant capacity of the extracts was evaluated by β-carotene/linoleic acid assay. Results showed that 60% of these extracts have antioxidant activity, exhibiting 50, 57% to 78, 96% inhibitions. While the inhibitory activity for oxidation of linoleic acid of 40% of them was less than 50%. From the present work it is possible to conclude that these microorganisms could be promising source of bioactive compounds, and warrant further study.

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

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.

In Vitro α-Glucosidase Inhibition and Antioxidative Potential of an Endophyte Species (Streptomyces sp. Loyola UGC) Isolated from Datura stramonium L

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±1.33μg/ml), scavenging activity on 2,2-diphenyl-picrylhydrazyl (DPPH) (IC50 435.31±1.79μg/ml), hydroxyl radical (IC50 350.21±1.02μg/ml), nitric oxide scavenging (IC50 800.12±1.05μg/ml), superoxide anion radical (IC50 220.31±1.47μg/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 176mg of catechol equivalents/gram extract. Perusal of this study indicates MeEA can be used as natural resource of α-glucosidase inhibitor and antioxidants.

Antibacterial polyketides from the marine alga-derived endophitic Streptomyces sundarbansensis: a study on hydroxypyrone tautomerism.

Polyketide 13 [=2-hydroxy-5-((6-hydroxy-4-oxo-4H-pyran-2-yl)methyl)-2-propylchroman-4-one] and three related known compounds 7, 9 and 11 were obtained and structurally characterized from Streptomyces sundarbansensis strain, an endophytic actinomycete isolated from the Algerian marine brown algae Fucus sp. Compound 13 was obtained as the major metabolite from optimized culture conditions, by using Agar state fermentation. Due to tautomeric equilibrium, 13 in CD3OD solution was able to incorporate five deuterium atoms, as deduced by NMR and ESI-MS/MS analysis. The 2-hydroxy-γ-pyrone form was established for these metabolites based on the comparison of their experimental IR spectra with the DFT calculated ones, for both the corresponding 4-hydroxy-α-pyrone and 2-hydroxy-γ-pyrone forms. During antibacterial evaluation, compound 13 stood out as the most active of the series, showing a selective activity against the gram positive pathogenic methicillin-resistant S. aureus (MRSA, MIC = 6 μΜ), with a bacteriostatic effect.

Actinomycetes: Role in Biotechnology and Medicine

Actinomycetes, one of the most diverse groups of filamentous bacteria, are well recognized for their metabolic versatility. The bioactive potential of these bacteria facilitates their survival even in distress and unfavourable ecological conditions. This special issue is dedicated to the importance of multitude of primary and secondary metabolites produced by actinomycetes. The six articles published in this issue balance the biocatalytic and biocidal potential of actinomycetes. The importance of large repertory of enzymes from actinomycetes and their potential in replacing chemical catalysts is discussed. Successful commercialization of these enzymes is an important step towards revolutionizing “green technology.” Reduction in the cost of enzyme production is demonstrated by production of endoglucanases from Streptomyces sp. on low-cost substrates. Such low-cost production initiatives can be extended to other enzymes and metabolites. Novel properties like thermal and ionic stabilities and a better turnover make these systems infallible and regenerative. The activity of enzymes from actinomycetes is not confined to substrate conversion alone but broadened to biocontrol of quorum-sensing-dependent phytopathogens, as mediated by acyl-homoserine-lactone-degrading enzymes from endophytic actinomycetes. Unexplored environments often appeal to researchers in the hope of accruing novel bacteria, a continuous quest which has actually led to discovery of unusually industrious microbes. Antimicrobial potential of actinobacteria isolated from the integument of Trachymyrmex fungus-growing ants is on par with commercial antimicrobials, clearly manifesting a new explorable niche “actinobacterial symbionts of plants and animals.” The term “antimicrobials” often leads our thoughts to “medicine-related” but it's “environment-related” applications are less contrived. Streptomyces lunalinharesii produces antimicrobial substances against sulfate-reducing bacteria commonly responsible for corrosion in the petroleum industry, with an ability to replace the existing biocides. Making the best out of the already good can be achieved for actinomycetes by strain improvement. Advanced microarray-driven reverse engineering strategies for the understanding and modulation of independently functioning regulatory pathways can allow these microfactories to overproduce important antibiotics. In a nutshell, actinomycetes offer the most promising synthesizers of many industrially and commercially meaningful metabolites. Novel and unexplored habitats may offer bacterial assemblages not reached hitherto. An integration of newer habitats, screening, and improvement technologies can offer promising candidates for biotechnology and health-related applications. Neelu Nawani Bertrand Aigle Abul Mandal Manish Bodas Sofiane Ghorbel Divya Prakash

Endophytic Actinomycetes: A Novel Source of Potential Acyl Homoserine Lactone Degrading Enzymes

Several Gram-negative pathogenic bacteria employ N-acyl-L-homoserine lactone (HSL) quorum sensing (QS) system to control their virulence traits. Degradation of acyl-HSL signal molecules by quorum quenching enzyme (QQE) results in a loss of pathogenicity in QS-dependent organisms. The QQE activity of actinomycetes in rhizospheric soil and inside plant tissue was explored in order to obtain novel strains with high HSL-degrading activity. Among 344 rhizospheric and 132 endophytic isolates, 127 (36.9%) and 68 (51.5%) of them, respectively, possessed the QQE activity. The highest HSL-degrading activity was at 151.30 ± 3.1 nmole/h/mL from an endophytic actinomycetes isolate, LPC029. The isolate was identified as Streptomyces based on 16S rRNA gene sequence similarity. The QQE from LPC029 revealed HSL-acylase activity that was able to cleave an amide bond of acyl-side chain in HSL substrate as determined by HPLC. LPC029 HSL-acylase showed broad substrate specificity from C6- to C12-HSL in which C10HSL is the most favorable substrate for this enzyme. In an in vitro pathogenicity assay, the partially purified HSL-acylase efficiently suppressed soft rot of potato caused by Pectobacterium carotovorum ssp. carotovorum as demonstrated. To our knowledge, this is the first report of HSL-acylase activity derived from an endophytic Streptomyces.

Micromonospora sonneratiae sp. nov., isolated from a root of Sonneratia apetala

A novel endophytic actinomycete, strain 274745(T), was isolated from a root of Sonneratia apetala collected in a mangrove forest in Sanya, Hainan province, China. The 16S rRNA gene sequence of strain 274745(T) showed the greatest similarity to Micromonospora pattaloongensis TJ2-2(T) (98.3 %). Phylogenetic analysis based on the gyrB gene also supported the close relationship of these two strains. The predominant menaquinone was MK-10(H8) and the major fatty acids were iso-C15 : 0, C17 : 0 and anteiso-C15 : 0. The characteristic whole-cell sugars were xylose and mannose. The cell wall contained meso-diaminopimelic acid and glycine. The polar lipid profile mainly comprised phosphatidylethanolamine, phosphatidylinositol and diphosphatidylglycerol. The DNA G+C content was 71.6 mol%. Furthermore, a combination of DNA-DNA relatedness and some physiological and biochemical properties indicated that the novel strain could be readily distinguished from the closest phylogenetic relatives. On the basis of these phenotypic and genotypic data, strain 274745(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora sonneratiae sp. nov. is proposed. The type strain is 274745(T) ( = CCTCC AA 2012003(T) = DSM 45704(T)).

Modestobacter roseus sp. nov., an endophytic actinomycete isolated from the coastal halophyte Salicornia europaea Linn., and emended description of the genus Modestobacter

A novel actinomycete, designated strain KLBMP 1279(T), was isolated from surface-sterilized roots of a coastal halophyte, Salicornia europaea Linn., collected from Jiangsu Province, in the east of China. The taxonomic status of this organism was established using a polyphasic approach. 16S rRNA gene sequence analysis indicated that strain KLBMP 1279(T) was closely related to Modestobacter marinus 42H12-1(T) (99.5% 16S rRNA gene sequence similarity), Modestobacter versicolor CP153-2(T) (98.4%) and Modestobacter multiseptatus AA-826(T) (97.5%). Chemotaxonomic characteristics were consistent with its assignment to the genus Modestobacter in that the isolate had meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall, MK-9(H4) as major menaquinone and a polar lipid profile containing diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, two unknown aminophospholipids and an unknown phospholipid. The predominant fatty acids were iso-C16:0, iso-C15:0 and C17:1ω8c. The DNA G+C content was 71.7 mol%. However, DNA-DNA hybridization assays as well as physiological and biochemical analyses differentiated strain KLBMP 1279(T) from its closest phylogenetic relatives. On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, the isolate KLBMP 1279(T) represents a novel species of the genus Modestobacter, for which the name Modestobacter roseus sp. nov. is proposed; the type strain is KLBMP 1279(T) (=KCTC 19887(T)=NBRC 108673(T)=DSM 45764(T)). An emended description of the genus Modestobacter is also proposed.

Xiangella phaseoli gen. nov., sp. nov., a member of the family Micromonosporaceae

A novel endophytic actinomycete, designated strain NEAU-J5(T) was isolated from roots of snap bean (Phaseolus vulgaris L.). Comparative analysis of the 16S rRNA gene sequence indicated that NEAU-J5(T) is phylogenetically related to members of the family Micromonosporaceae. The whole-cell sugars were galactose, mannose and glucose. The predominant menaquinones were MK-9(H4) and MK-9(H6). The major fatty acids were C16:0, C18:0, C17:1ω7c, iso-C15:0 and C17:0. The phospholipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. The DNA G+C content was 72.2 mol%. On the basis of the morphological and chemotaxonomic characteristics, phylogenetic analysis and characteristic patterns of 16S rRNA gene signature nucleotides, strain NEAU-J5(T) represents a novel species of a new genus within the family Micromonosporaceae, for which the name Xiangella phaseoli gen. nov., sp. nov. is proposed. The type strain of Xiangella phaseoli is strain NEAU-J5(T) (=CGMCC 4.7038(T)=DSM 45730(T)).

Amycolatopsis jiangsuensis sp. nov., a novel endophytic actinomycete isolated from a coastal plant in Jiangsu, China

A novel actinomycete, designated strain KLBMP 1262(T), was isolated from a coastal plant Dendranthema indicum (Linn.) Des Moul collected from the coastal region of Nantong, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP 1262(T) is a member of the genus Amycolatopsis. The 16S rRNA gene sequence similarity indicated that strain KLBMP 1262(T) is most closely related to Amycolatopsis sulphurea DSM 46092(T) (97.96 %), Amycolatopsis ultiminotia RP-AC36(T) (97.50 %) and Amycolatopsis jejuensis N7-3(T) (97.44 %); similarity to other type strains of the genus Amycolatopsis was less than 97.0 %. The organism was determined to have chemical and morphological features consistent with its classification in the genus Amycolatopsis such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was determined to be MK-9 (H(4)). The polar lipids detected were phosphatidylmethylethanolamine, phosphatidylethanolamine, an unknown aminophospholipid, two unknown glycolipids and several unknown lipids. The major fatty acids were found to be C(16:0), iso-C(16:0) and iso-C(15:0). DNA-DNA relatedness data, together with phenotypic differences, clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate is considered to represent a novel species, for which the name A. jiangsuensis sp. nov. is proposed. The type strain is KLBMP 1262(T) (=KCTC 19885 (T) = NBRC 108679(T)).

Wangella harbinensis gen. nov., sp. nov., a new member of the family Micromonosporaceae

A novel endophytic actinomycete, designated strain NEAU-J3(T), was isolated from soybean root (Glycine max (L.) Merr) and characterized using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain NEAU-J3(T) fell within the family Micromonosporaceae. The strain was observed to form an extensively branched substrate mycelium, which carried non-motile oval spores with a smooth surface. The cell walls of strain NEAU-J3(T) were determined to contain meso-diaminopimelic acid and galactose, ribose and glucose were detected as whole-cell sugars. The major menaquinones were determined to be MK-9(H(4)) and MK-9(H(6)). The phospholipids detected were phosphatidylcholine and phosphatidylethanolamine. The major cellular fatty acids were determined to be C(16:0), C(18:1) ω9c, C(18:0), C(17:0), C(17:1) ω7c, anteiso-C(17:0), C(16:1) ω7c and C(15:0). The DNA G + C content was 62.5 mol%. On the basis of the morphological and chemotaxonomic characteristics, phylogenetic analysis and characteristic patterns of 16S rRNA gene signature nucleotides, strain NEAU-J3(T) is considered to represent a novel species of a new genus within the family Micromonosporaceae, for which the name Wangella harbinensis gen. nov., sp. nov. is proposed. The type strain of Wangella harbinensis is strain NEAU-J3(T) (=CGMCC 4.7039(T) = DSM 45747(T)).

Bioprospecting lignocellulolytic enzymes from endophytic actinomycetes aiming at second generation ethanol production

Bioprospecting lignocellulolytic enzymes from endophytic actinomycetes aiming at second generation ethanol production

Plant growth enhancing effects by a siderophore-producing endophytic streptomycete isolated from a Thai jasmine rice plant (Oryza sativa L. cv. KDML105)

An endophytic Streptomyces sp. GMKU 3100 isolated from roots of a Thai jasmine rice plant (Oryza sativa L. cv. KDML105) showed the highest siderophore production on CAS agar while phosphate solubilization and IAA production were not detected. A mutant of Streptomyces sp. GMKU 3100 deficient in just one of the plant growth promoting traits, siderophore production, was generated by inactivation of a desD-like gene encoding a key enzyme controlling the final step of siderophore biosynthesis. Pot culture experiments revealed that rice and mungbean plants inoculated with the wild type gave the best enhancement of plant growth and significantly increased root and shoot biomass and lengths compared with untreated controls and siderophore-deficient mutant treatments. Application of the wild type in the presence or absence of ferric citrate significantly promoted plant growth of both plants. The siderophore-deficient mutant clearly showed the effect of this important trait involved in plant-microbe interaction in enhancement of growth in rice and mungbean plants supplied with sequestered iron. Our results highlight the value of a substantial understanding of the relationship of the plant growth promoting properties of endophytic actinomycetes to the plants. Endophytic actinomycetes, therefore, can be applied as potentially safe and environmentally friendly biofertilizers in agriculture.

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

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.

Genes expressed in tissue-cultured seedlings of mountain laurel (Kalmia latifolia L.) with colonizing Streptomyces padanus AOK30

An endophytic actinomycete, Streptomyces padanus AOK30, is capable of protecting mountain laurel against infection by Pestalotiopsis sydowiana, a causal agent of Pestalotia disease, when applied to the seedling of the plant. In this study, suppression subtractive hybridization was used to identify genes differentially expressed in seedlings of mountain laurel after application of S. padanus AOK30. Subsequent dot hybridization with independent RNA from S. padanus-colonized and control plants identified nonredundant 180 cDNAs involving 71 and 109 clones, which were up- and downregulated after inoculation with the bacteria, respectively. Comparison of the sequences with databases revealed that a number of transcripts encoding proteins or enzymes that function directly in defense or stress response and regulatory proteins were regulated differentially in the seedlings with colonizing S. padanus AOK30. Semi-quantitative RT-PCR analysis for the selected genes demonstrated that inoculation of mountain laurel seedlings with S. padanus AOK30 increased expression of defense-related genes as well as distinct classes of glutathione S-transferase, although endochitinases were exclusively suppressed. These results clearly indicate that the S. padanus-colonizing seedlings likely initiate or prime plant defense responses toward pathogen infection. Selected genes were also differentially expressed in S. padanus-colonized seedlings, compared to those solely challenged with the fungal pathogen P. sydowiana. This approach will assist in efforts not only to understand the molecular basis of the enhanced tolerance and/or enhanced disease resistance of mountain laurel, but also to define a core set of genes during colonization or association with S. padanus AOK30.

Characterisation of endophytic actinomycetes isolated from wattle trees (Acacia auriculiformis A. Cunn. ex Benth.) in Thailand

Eleven strains of endophytic actinomycetes were isolated from healthy roots of Acacia auriculiformis A. Cunn . ex Benth. collected from Bangkok and Nakhonpathom, Thailand. No actinomycete could be isolated from leaf samples. Analysis of 16S rRNA sequencing of those strains revealed that they belong to members of genera Streptomyces , Actinoallomurus , Amycolatopsis , Kribbella and Microbispora . Within 11 endophytic actinomycetes, GMKU 944 showed strongest antibacterial activities against tested bacteria and fungi. In this work, the residing property of endophytic actinomycetes in the roots of wattle tree was verified by inoculation of GMKU 937 to germinated seeds. It was demonstrated that GMKU 937 could be re-isolated and was a true endophyte.

Overproduction and biological activity of prodigiosin-like pigments from recombinant fusant of endophytic marine Streptomyces species

Thirty-four endophytic marine Actinomycetes isolates were recovered from the Egyptian marine sponge Latrunculia corticata, out of them 5 isolates (14.7%) showed red single colonies on yeast-CzAPEK plates. Isolates under the isolation code NRC50 and NRC51 were observed with the strongest red biomass. After application of protoplast fusion between NRC50 and NRC51 isolates, 26 fusants were selected and produced widely different amounts of prodigiosin-like pigments (PLPs) on different fermentation media. Among them fusant NRCF69 produced 79 and 160.4% PLPs more than parental strains NRC50 and NRC51, respectively. According to the analysis of 16S rDNA sequence (amplified, sequenced, and submitted to GenBank under Accession no. JN232405 and JN232406, respectively), together with their morphological and biochemical characteristics, parental strains NRC50 (P1) and NRC51 (P2) were identified as Streptomyces sp. and designated as Streptomyces sp. NRC50 and Streptomyces sp. NRC51. This study describes a low cost, effective production media by using peanut seed broth, sunflower oil broth or dairy processing wastewater broth alone, or supplemented with 0.5% mannitol that supports the production of PLPs by the Streptomyces fusant NRCF69 under study (42.03, 40.11, 36.7 and 47gL(-1), respectively). PLPs compounds exhibited significant cytotoxic activities against three human cancer cell lines: colon cancer cell line (HCT-116), liver cancer cell line (HEPG-2) and breast cancer cell line (MCF-7) and antimycotic activity against clinical dermatophyte isolates of Trichophyton, Microsporum and Epidermophyton.