Isolation,characterisation and antifungal activity of marine actinobacteria from Goa and Kerala,the west coast of India

In total, 53 marine actinobacteria were isolated from the soils of six different locations in Goa and Kerala, on the west coast of India. All the isolates were screened for their antifungal properties against some phytopathogenic fungi by dual culture experiments. Among the 53 actinobacterial isolates, five isolates inhibited the growth of phytopathogens, namely Colletotrichum falcatum, Thielaviopsis paradoxa and Fusarium semitectum. But none of them were effective against Aspergillus niger, Aspergillus candidus and Aspergillus flavus. The antifungal activity of the actinobacteria was tested by food poisoning techniques, using four different concentrations (0.5%, 1.0%, 1.5% and 2.0%) of cell-free culture filtrates, which showed promising activity (almost 100% inhibition) against three pathogenic and one non-pathogenic fungi at 2% extract concentration. A comparison of the antifungal activity of the actinobacteria was also made with three commercial fungicides, namely hexaconazole, thiophanate methyl and propiconazole. The identity of the antagonistic actinobacteria was confirmed based on the morphological, cultural, biochemical, chemo-taxonomical and physiological characteristics. Among 5 antagonistic isolates, three antagonistic isolates were assigned to the genus Streptomyces, Nocardiopsis (1) and Saccharopolyspora (1).     

Greek indigenous streptomycetes as biocontrol agents against the soil‐borne fungal plant pathogen Rhizoctonia solani

Aims

To examine the biocontrol potential of multiactive Greek indigenous Streptomyces isolates carrying antifungal activity against Rhizoctonia solani that causes damping‐off symptoms on beans.

Methods and Results

A total of 605 Streptomyces isolates originated from 12 diverse Greek habitats were screened for antifungal activity against R. solani DSM843. Almost one‐third of the isolates proved to be antagonistic against the fungus. From the above isolates, six were selected due to their higher antifungal activity, identified by analysing their 16S rRNA gene sequence and studied further. The obtained data showed the following: firstly, the isolates ACTA1383 and ACTA1557 exhibited the highest antagonistic activity, and therefore, they were selected for in vivo experiments using bean seeds as target; secondly, in solid and liquid culture experiments under optimum antagonistic conditions, the medium extracts from the isolates OL80, ACTA1523, ACTA1551 and ACTA1522 suppressed the growth of the fungal mycelium, while extracts from ACTA 1383 and ACTA1557 did not show any activity.

Conclusions

These results corresponded important indications for the utility of two Greek indigenous Streptomyces isolates (ACTA1557 and ACTA1383) for the protection of the bean crops from R. solani damping‐off symptoms, while four of them (isolates OL80, ACTA1523, ACTA1551 and ACTA1522) seem to be promising producers of antifungal metabolites.

Significance and Impact of the Study

This is the first study on the biocontrol of R. solani using multiactive Streptomyces isolates originated from ecophysiologically special Greek habitats. Our study provides basic information to further explore managing strategies to control this critical disease.     

海南东寨港真红树植物内生放线菌多样性及其抗菌活性

【目的】勘探海南东寨港真红树植物内生放线菌多样性,为发现放线菌新物种和新抗生素奠定基础。【方法】样品经表面消毒后粉碎,用10种不同培养基分离放线菌;通过PCR扩增、测定并比对16S r RNA基因序列,开展放线菌多样性分析;通过发酵、萃取等处理方法得到四类样品,包括发酵原液、乙酸乙酯提取液及水层和菌体的丙酮浸泡提取液;采用纸片扩散法对样品进行抗菌活性筛选;基于PCR的基因筛选技术探测活性菌株可能存在的NRPS、PKS I、PKS II抗生素生物合成基因。【结果】经形态特征排重,从14种真红树植物样品中共得到放线菌146株,16S r RNA基因序列比对表明它们分布于13个科18个属,其中链霉菌属为优势菌属,菌株S3Cf-2和S3Af-1的16S r RNA基因序列分别与有效发表菌株Couchioplanes caeruleus DSM44103T(X93202)和Microlunatus terrae BS6T(JF806519)的相似率最高,分别为97.45%和97.43%,可能为新物种。对其中46株放线菌发酵样品的抗菌活性检测表明,40株具有抗菌活性,总阳性率为86.96%;活性菌株中,38株菌存在至少一种所探测的生物合成基因簇,阳性率为95%,其中14株同时具有所探测的3种抗生素生物合成基因簇。【结论】海南东寨港真红树植物中存在多样性丰富的药用放线菌资源,具有从中发现放线菌新物种及新抗生素的潜力。     

亚热带红壤区森林土壤剖面微生物残体碳分布及影响因素

土壤剖面中20cm以下土壤有机碳(SOC)储量占土壤剖面总SOC储量50%左右,由于土壤微生物残体碳(MRC)是稳定土壤碳库的重要来源,因此研究土壤剖面中MRC对SOC的贡献对于评估土壤碳储量具有重要意义。然而,目前关于MRC含量及其对SOC贡献的研究多数集中在土壤表层,在土壤剖面和母质中尚不清楚。选取江西省千烟洲亚热带典型森林红壤剖面,通过氨基糖与磷脂脂肪酸(PLFA)微生物标志物分析方法,分析红壤剖面和母质中MRC的影响机制及其对SOC贡献的分布特征。研究结果表明：(1)MRC含量随着土壤剖面深度增加而显著降低(P<0.05),在整个土壤剖面中,细菌MRC对SOC贡献为6%—12%,真菌MRC对SOC贡献为12%—36%,MRC对SOC贡献为18%—46%。从土壤表层至母质,真菌MRC对SOC贡献高于细菌MRC。(2)结构方程模型结果表明,在土壤剖面中,MRC含量主要受到微生物-PLFA含量、容重和溶解态有机碳含量的影响。研究量化了红壤剖面中MRC对SOC的贡献,表明在20cm以下土壤及母质中,微生物残体碳对红壤地区生态系统碳库具有重要贡献。     

Research on marine actinobacteria in India

Marine actinobacteriology is one of the major emerging areas of research in tropics. Marine actinobacteria occur on the sediments and in water and also other biomass (mangrove) and substrates (animal). These organisms are gaining importance not only for their taxonomic and ecological perspectives, but also for their unique metabolites and enzymes. Many earlier studies on these organisms were confined only to the temperate regions. In tropical environment, investigations on them have gained importance only in the last two decades. So far, from the Indian peninsula, 41 species of actinobacteria belonging to 8 genera have been recorded. The genus, Streptomyces of marine origin has been more frequently recorded. Of 9 maritime states of India, only 4 have been extensively covered for the study of marine actinobacteria. Most of the studies conducted pertain to isolation, identification and maintenance of these organisms in different culture media. Further, attention has been focused on studying their antagonistic properties against different pathogens. Their biotechnological potentials are yet to be fully explored.     

Pharmaceutically active secondary metabolites of marine actinobacteria

Marine actinobacteria are one of the most efficient groups of secondary metabolite producers and are very important from an industrial point of view. Many representatives of the order Actinomycetales are prolific producers of thousands of biologically active secondary metabolites. Actinobacteria from terrestrial sources have been studied and screened since the 1950s, for many important antibiotics, anticancer, antitumor and immunosuppressive agents. However, frequent rediscovery of the same compounds from the terrestrial actinobacteria has made them less attractive for screening programs in the recent years. At the same time, actinobacteria isolated from the marine environment have currently received considerable attention due to the structural diversity and unique biological activities of their secondary metabolites. They are efficient producers of new secondary metabolites that show a range of biological activities including antibacterial, antifungal, anticancer, antitumor, cytotoxic, cytostatic, anti-inflammatory, anti-parasitic, anti-malaria, antiviral, antioxidant, anti-angiogenesis, etc. In this review, an evaluation is made on the current status of research on marine actinobacteria yielding pharmaceutically active secondary metabolites. Bioactive compounds from marine actinobacteria possess distinct chemical structures that may form the basis for synthesis of new drugs that could be used to combat resistant pathogens. With the increasing advancement in science and technology, there would be a greater demand for new bioactive compounds synthesized by actinobacteria from various marine sources in future.     

Chemical Constituents from a Mangrove-Derived Actinobacteria Isoptericola chiayiensis BCRC 16888 and Evaluation of Their Anti-NO Activity

Cultivation of the actinobacteria strain Isoptericola chiayiensis, a mangrove - derived actinobacteria that was isolated from a mangrove soil collected in Chiayi County, resulted in the isolation of one new 2-furanone derivative, isopterfuranone ( 1 ), one new sesquiterpenoid, isopterchiayione ( 2 ), one new benzenoid derivative, isopterinoid ( 3 ), five new flavonoids, chiayiflavans A−E ( 4 – 8 ), and 4 metabolites isolated for the first time from nature source, methyl 3-(4-methyl-2,5-dioxopyrrolidin-3-yl)propanoate ( 9 ), 3-ethyl-4-methylpyrrolidine-2,5-dione ( 10 ), chiayiensol ( 11 ) and chiayiensic acid ( 12 ). Their structures were determined through in-depth spectroscopic and mass-spectrometric analyses. Most of the isolates showed potent inhibitory effects on NO production in LPS-stimulated RAW 264.7 murine macrophages cells with IC₅₀ values ranging from 9.36 to 40.02 μM. Of these isolates, 4 and 5 showed NO inhibitory activity with IC₅₀ values of 17.14 and 9.36 μM, stronger than the positive control quercetin (IC₅₀=36.95 μM). This is the first report on flavan metabolites from the genus Isoptericola.     

Characterization of some efficient cellulase producing bacteria isolated from paper mill sludges and organic fertilizers

The wide variety of bacteria in the environment permits screening for more efficient cellulases to help overcome current challenges in biofuel production. This study focuses on the isolation of efficient cellulase producing bacteria found in organic fertilizers and paper mill sludges which can be considered for use in large scale biorefining. Pure isolate cultures were screened for cellulase activity. Six isolates: S1, S2, S3, S4, E2, and E4, produced halos greater in diameter than the positive control (Cellulomonas xylanilytica), suggesting high cellulase activities. A portion of the 16S rDNA genes of cellulase positive isolates were amplified and sequenced, then BLASTed to determine likely genera. Phylogenetic analysis revealed genera belonging to two major Phyla of Gram positive bacteria: Firmicutes and Actinobacteria. All isolates were tested for the visible degradation of filter paper; only isolates E2 and E4 (Paenibacillus species) were observed to completely break down filter paper within 72 and 96 h incubation, respectively, under limited oxygen condition. Thus E2 and E4 were selected for the FP assay for quantification of total cellulase activities. It was shown that 1% (w/v) CMC could induce total cellulase activities of 1652.2±61.5 and 1456.5±30.7 μM of glucose equivalents for E2 and E4, respectively. CMC could induce cellulase activities 8 and 5.6X greater than FP, therefore CMC represented a good inducing substrate for cellulase production. The genus Paenibacillus are known to contain some excellent cellulase producing strains, E2 and E4 displayed superior cellulase activities and represent excellent candidates for further cellulase analysis and characterization.     

Optimization of extracellular endoxylanase, endoglucanase and peroxidase production by Streptomyces sp. F2621 isolated in Turkey

AIMS: To determine the effect of environmental conditions on the production of extracellular lignocellulose-degrading enzymes by Streptomyces sp. F2621 and to assess the potential use of these enzymes in the hydrolysis of lignocellulose material. METHODS AND RESULTS: The production of extracellular lignocellulose-degrading enzymes, endoxylanase, endoglucanase and peroxidase during the growth of Streptomyces sp. F2621 in basal salts-yeast extract medium containing different carbon sources and the effect of a number of environmental parameters (e.g. carbon sources and concentrations, pH and temperature) were investigated. The highest endoxylanase (22.41 U ml(-1)) and peroxidase (0.58 U ml(-1)) activities were obtained after 2-4 days of incubation at 30 degrees C in a basal salts medium containing 0.4% (w/v) oat spelt xylan and 0.6% (w/v) yeast extract, corresponding to C : N ratio of 6 : 1. Cell-free extracellular enzyme preparations from the strain were capable of releasing both sugar and aromatic compounds during incubation with eucalyptus paper pulp, straw and xylan. Overall, 9.3% hydrolysis of xylan occurred after 24-h incubation. However the rates of hydrolysis of paper pulp and straw were approximately twofold less than xylan hydrolysis, although the total percentage hydrolysis of available substrate (24.5% and 16.3%, respectively) was greater than xylan hydrolysis. CONCLUSIONS: The high levels of enzyme production achieved under batch cultivation conditions, coupled with no significant production of endoglucanase during the growth phase of organism and the release of both sugar and aromatic compounds from paper pulp and straw signify the suitability for these enzymes for industrial applications such as pulp and paper production. SIGNIFICANCE AND IMPACT OF THE STUDY: The results highlight the environmental conditions for the production of extracellular lignocellulose-degrading enzymes by Streptomyces sp. F2621 and suggest the use of streptomycetes and/or their enzymes in industrial processes.     

The potential of antagonistic moroccan Streptomyces isolates for the biological control of damping‐off disease of pea (Pisum sativum L.) caused by Aphanomyces euteiches

Three hundred and fifty‐nine isolates of actinobacteria collected from different Moroccan soils were evaluated for their in vitro antimicrobial activity against the oomycete pathogen Aphanomyces euteiches, the causal agent of damping‐off of pea and other legumes. Eighty‐seven isolates (24%) had an inhibitory in vitro effect against A. euteiches. Fourteen bioactive isolates with the greatest inhibitory effect against A. euteiches and no inhibitory effect on plant beneficial rhizobia were tested for their ability to protect pea seeds and seedlings against the damping‐off disease using culture supernatants or spore suspensions as treatments. The two most protective isolates, OB21 and BA15, significantly reduced, compared to untreated control plants, damping‐off by 33% and 47%, respectively. The two bioactive isolates were classified as species of the genus Streptomyces based on 16S rDNA analysis and morphological and chemical characteristics.