Ann Microbiol (2014) 64:1829–1838 DOI 10.1007/s13213-014-0831-1
ORIGINAL ARTICLE
Isolation and characterization of novel alkali-halophilic actinomycetes from the Chilika brackish water lake, India Nityanand Malviya & Mahesh S. Yandigeri & Arvind Kumar Yadav & Manoj Kumar Solanki & Dilip K. Arora
Received: 1 April 2013 / Accepted: 3 February 2014 / Published online: 20 February 2014 # Springer-Verlag Berlin Heidelberg and the University of Milan 2014
Abstract The current investigation deals with isolation and functional characterization of agriculturally important novel alkali-halophilic actinomycetes from water and sediment samples of three distinct niches, namely, central, south and sea mouth sectors of the Chilika brackish water lake. A total of 59 different morphotypes based on phenotypic characterization were isolated from different sectors of the lake. Among them, a total of twenty one isolates were screened as alkalihalophiles based on screening for salt and pH tolerance on starch casein agar medium amended with 10 % NaCl and at pH 9.0. Further characterization for plant growth promotion and biocontrol attributes revealed that the south sector harboured a maximum percentage of siderophore producers while the central sector had the highest indole 3-acetic acid as well as extracellular protease producers. The sea mouth sector had a higher portion of actinomycetes possessing nitrate reductase activity and some biocontrol attributes such as antimicrobial activity against fungal pathogens (Rhizoctonia solani, Fusarium udum and Fusarium oxysporum f. sp. ciceri) and production of chitinase enzyme. Identification including 16S rRNA gene sequences revealed that isolates belonged to the Streptomyces and Micromonospora genera, respectively. Keywords Actinomycetes . Chilika Lake . PGP traits . 16S rRNA gene Electronic supplementary material The online version of this article (doi:10.1007/s13213-014-0831-1) contains supplementary material, which is available to authorized users. M. S. Yandigeri (*) National Bureau of Agriculturally Important Insects, P.B. No. 2491, H.A. Farm Post, Bellary Road, Bangalore 560032, Karnataka, India e-mail:
[email protected] N. Malviya : A. K. Yadav : M. K. Solanki : D. K. Arora National Bureau of Agriculturally Important Microorganisms, Kusmaur, Kaithauli, Mau Nath Bhanjan 275101, India
Introduction Microorganisms can colonize in various ecological niches including extremophilic habitats due to their adaptive features including structural as well as functional adaptations (Jiang et al. 2006). Chilika brackish water lake represents one of such niches and studies of microbial diversity from various saline habitats have been previously reported (Rao et al. 2000; Litchfield and Gillevet 2002; Mancinelli 2005), but actinobacterial diversity of saline lakes was lacking. The current study focussed on Chilika lake, which covers both fresh water as well as marine ecotones and possesses hostile environments enriched with immense biodiverse values. Chilika lake is a pear shaped, ephemeral lake and is the biggest coastal lake in Asia. It is the second largest lake in the world, with a maximum length of 64.3 km and a width of 20.1 km. The lake was once a part of the Bay of Bengal and has a long history of more than 5,000 years. It is divided into North, South, Central and Sea mouth sectors and is fed by both marine (Bay of Bengal) as well as fresh waters rivers and rivulets such as the Daya, Nuna, Ratnachira, Bhargavi, Malguni, Dhanu and Salia. In 1981, Chilika Lake was designated the first Indian wetland of international importance under the Ramsar Convention in danger record (Dube et al. 2010) and is one of the best studied coastal lakes in the tropics. Actinomycetes are one of the major microbial dominant groups and are well known for their saprophytic behaviour as well as for production of diverse bioactive secondary metabolites. They are also recognised for their capacity to survive in extreme habitats (Bredholdt et al. 2007). There are very few reports available pertaining to alkali-halophilic actinomycetes diversity from lake ecosystems (Suthindhiran and Kannabiran 2009). Members of alkali-halophilic actinomycetes are not much explored and were poorly understood. There are only a few reports available pertaining to actinomycetes from saline and alkaline habitats. Recent findings from culturable and
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unculturable diversity have demonstrated that there are tremendous diversities and novelties among the halotolerant and alkaliphilic actinomycetes present in saline and alkaline environments (Chen et al. 2009; Luo et al. 2009). In the past three decades, discoveries of new compounds have declined from soil derived actinomycetes (Fu et al. 2011). For this reason, researchers have started focusing towards unexplored niches like marine sources, lagoons, lakes and saltpans. They are exciting novel sources of bioactive compounds with promising anti-microbial activity against pathogenic strains (Cao et al. 2004; Castillo et al. 2007). Actinomycetes produce large number of antimicrobial compounds and plant growth promoting hormones are reported. There are many references cited with an ability of the Streptomyces, Micromonospora, Corynebacterium, Frankia, Mycobacterium, and Rhodococcus (Tsavkelova et al. 2005; Khamna et al. 2010) genera in plant growth promotion directly (production of phytohormones) (Hopwood and Chater 1990; Houssam 2009) or indirectly (production of cell wall degrading enzymes) (Walter and Crawford 1995). In spite of this, streptomycetes is one of the dominant groups of microbes and plays a major role in environmental sustainability, while actinomycetes diversity in chilika lake is still untouched. This study focused on the differences of community structure of actinomycetes from sea water and fresh water as well as characterized potentialities of isolates for its agricultural and industrial importance. Morphological and chemotaxonomic characterization are useful aids in the identification of actinomycetes however, they may fail to correctly identify species of several genera that exhibit similar morphological and chemotaxonomic properties (Wang et al. 1996; Zhang et al. 1998). Polyphasic approaches including the16S rRNA gene sequence has been widely used to determine taxonomic positions of many organisms in virtually all taxonomic ranks (Wang et al. 1999). The present study aimed to investigate the distribution, phylogeny and functional attributes of alkali-halophilic actinomycetes isolated from Chilika brackish water lake with respect to plant growth promoting traits, hydrolytic enzymes and antifungal activity, respectively.
Materials and methods Site description, sampling and analysis Chilika lake (19°28′-19°54′ N and 85°06′-86°36′E) is situated on the east coast of India and topographic wind factors blow from South and South West directions from February to September. As the tidal effect appears negligible, wind acts as the dominant force. The sampling was carried out in the summer season June, 2009; from different zones/areas, namely, Central (Chadheiguha S1; Nalabana S2), South (Rambha
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S3; Badakuda S4), and marine source (Manika patana S5; and Sea Mouth S6) of Chilika lake (Fig. 1, Table 1). Random stratified water samples at a 0.5 m depth from the surface as well as sediment samples were collected in triplicates by using sterile bottles and a grab sampler under aseptic conditions. Collected samples were divided into two parts; one was used for the analysis of physiochemical properties and other for the isolation of actinomycetes. The pH and electrical conductivity were measured by a pH meter (Eutech Instruments, pH 1,500) and a EC meter at room temperature (28 °C) (Universal Bio, India), respectively. Isolation, screening and characterization of actinomycetes For isolation of different actinomycetes, lake sediment and water samples were subjected to pre-heat treatment prior to serial dilution and plating. The serial dilution was performed using Ringer’s solution. Pre-heat treatment was performed by incubating the water and sediment samples in a water bath at 50 °C for 60 min (Takizawa et al. 1993). We used enrichment techniques at 28 °C incubation temperatures, like CaCO3 enrichment (1 %) (Tsao et al. 1960), SDS (5 %) and phenol treatment (1.4 %) (Hayakawa and Nonomura 1989; Hayakawa et al. 2004) to favour growth of actinomycetes and inhibit/retard most fungi. Various media such as, starch casein agar (SCA), ISP media (1–7), humic acid vitamin agar (HVA) and actinomycetes isolation agar (AIA) amended with nystatin (25 mg ml−1), cycloheximide (50 mg ml−1) and nalidixic acid (50 mg ml−1) were employed to isolate Streptomyces and related genera (Pan et al. 2009). Morphological and physiological characterization was carried out by using standard protocols by using ISP media (Shirling and Gottlieb 1966). For the screening of alkali-halophilic nature of the isolates, they were evaluated by observing the growth at 28±2 °C in starch casein agar medium amended with different concentrations of NaCl (5–10 % w/v; pH 9.0). Plant growth promotion (PGP) traits like siderophore production, nitrate reduction and indole 3-acetic acid (IAA) production were assayed according to Malviya et al. (2011). Protease production and chitinase assay (Hsu and Lockwood 1975), antifungal activity against phytopathogenic fungi (collected from National Agriculturally Important Microbial Culture Collection, Mau Nath Bhanjan, U.P., India) such as Rhizoctonia solani (NAIMCC-F-01970), Fusarium udum (NAIMCC-F-01041) and F. oxysporum f. sp. ciceri (NAIMCC-F-00858) were also evaluated and analysed using a scanning electron microscope (Walter and Crawford 1995; Malviya et al. 2011). Molecular characterization of alkali-halophilic actinomycetes Actinomycetes genomic DNA was extracted using the modified method of Boudjella et al. (2006) and 16S rRNA gene
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Fig. 1 Map showing the sampling sites (water and sediment samples) from Chilika lake, India
amplification was performed using universal bacterial primers fD1 (5′-AGAGTTTGATCCTGGCTCAG-3′) and rP2 (5′AAGGAGGTGATCCAGCCGCA-3′) (Weisberg et al. 1991). The final volume of the reaction mixture of 50 μl contained 10× PCR buffer (10 mM Tris–HCl, 50 mM KCl, pH 9.0 at 25 °C), 1.5 mM MgCl2, 200 μM of each dNTP, 1 pM of each primer, 0.25U of Taq polymerase and 50 ng of template DNA. The amplification was performed on a BioRad thermal cycler (initial denaturation step at 98 °C for 3 min, followed by 30 amplification cycles of 94 °C for 60 s, 54 °C for 60 s, and 72 °C for 90 s) and was observed through horizontal electrophoresis. Purified amplified product of 21 alkali-halophilic strains was sequenced using Taq-mediated dideoxy chain terminator cycle in ABI 3130xl automated genetic analyser (Applied Biosystems, UK). Gaps were treated by pairwise deletion and bootstrap analysis was done by using 5,000 pseudoreplications. Data analysis The sequences generated by an automated genetic analyzer were compared with known sequences deposited in a public database by the Basic Local Alignment Search Tool online software (BLAST; http://www.ncbi.nlm.nih.gov/BLAST/),
and the nearest neighbor actinobacterial sequences were aligned with all sequences by using CLUSTAL W and phylogenetic analysis carried out with the MEGA software version 4.1 (Saitou and Nie 1987).
Results Isolation, screening and characterization of alkali-halophilic actinomycetes isolates A total of 59 different morphotypes, classified on the basis of color of aerial and substrate mycelium, pigmentation and microscopic examination obtained after utilizing different enrichment techniques and media, was obtained from different sectors of Chilika lake (Table 2, Fig. 2). Further, screening for the alkali-halophilic nature of 59 “morphotypes” revealed that, a total of 21 isolates possessed the ability to grow at pH 9.0 and within a concentration of 5 to 10 % NaCl on starch casein agar (Supplementary Table 1). An attempt to grow the isolates at more than pH 9.0 and 10 % NaCl failed in the case of actinomycetes, and we did not attempt to grow below pH 8.0, as the aim of the current study was mainly to isolate alkalihalophilic actinomycetes. The population frequency of
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33.04±0.20 32.90±0.29 S6 (Sea Mouth)
S5 (Manika patna)
S4 (Badakuda)
S3 (Rambha)
S2 (Nalabana)
Water Sediment Total=
8.24±0.21 8.16±0.20
3 5 116
2 4 59
1 3 21
– S1CS1 S2NW2 S2NS5 S3RW3 S3RS5, S3RS8, S3RS9 S4BW2 S4BS3, S4BS5, S4BS6 S5MW2 S5MS1, S5MS2, S5MS3, S5MS6, S5MS7 S6SW1 S6SS1, S6SS2, S6SS3 0 1 1 1 1 3 1 3 1 5 5 9 5 8 3 8 2 5 2 6 4.09±0.21 4.11±0.33 12.82±0.21 12.70±0.24 13.36±0.24 13.01±0.31 13.06±0.21 12.97±0.22 30.56±0.29 29.97±0.12 Water Sediment Water Sediment Water Sediment Water Sediment Water Sediment S1 (Chadheiguha)
8.21±0.07 8.02±0.08 8.67±0.12 8.27±0.05 8.44±0.20 8.29±0.09 8.38±0.34 8.23±0.07 8.34±0.22 8.26±0.25
10 23 9 18 7 17 6 6 5 7
Number of alkali-halophilic “morphotypes” Number of “morphotypes” Number of colonies/ composite sample
Population count (number of colonies×103 per gram or per mL of sample) Salinity (%) pH Sampling material Site denomination
Table 1 Characterization of the collection sites, enumeration of actinomycetes and screening of alkali-halophilic actinomycetes from Chilika lake, India
Alkali-halophilic strain code
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actinomycetes isolated per sample in Chilika lake was shown to be different in all three sectors. The central sector harboured a maximum population frequency (Chadheiguha 28.4 % and Nalabana 23.2 %), followed by the south sector (Rambha, 21.5 %; Badakuda, 10.3 %), while the least was recorded in the sea mouth sector (Manika patana, 10.3 %; sea mouth, 6.8 %). Sediment samples had higher actinomycetes population frequencies than lake water while the population frequency of alkali-halophilic actinomycetes (pH 9.0; NaCl 10 %) decreased as we shifted away from marine habitat to the fresh water lake (Table 1, Fig. 3). Preliminary identification of selected isolates was performed based on morphological characterization using aerial and substrate mycelia color, pigments, and spore chain arrangements were assigned as they belong to Streptomyces and Micromonospora genera. Functional characterization of all the isolates revealed that the south sector harboured a maximum percentage of siderophore producers (48.5 %) while the central sector had the highest IAA (45 %), and extracellular protease enzyme (45.1 %) producers. The sea mouth sector was enriched with isolates having nitrate reductase activity (42.3 %) and had biocontrol attributes as well as antimicrobial activity (38.8 %) and possessed machinery for chitinase enzyme production (39 %). Chitinase and protease enzyme producing isolates were procured in a majority of sediment samples while cellular siderophores, IAA, antimicrobial activity and nitrate reductase potential was found to be highest in water samples (Fig. 4). Interaction studies of isolates with phytopathogenic fungi captured using scanning electron microscopy showed rupturing of fungal mycelium, colonization and finally complete destruction (Fig. 5, Supplementary Table 2). The catabolic carbon assimilation pattern was analysed based on utilization/non-utilization of 95 substrates studied by the BIOLOG™ system, and all actinomycetes isolates showed different types of carbon substrate utilization patterns ranging from 9 to 82 out of 95 substrates. Among all of the isolates, the marine sector isolates possessed a higher substrate utilization pattern. Isolates from sediment samples utilized more carbon substrate than water derived isolates and isolates from the south sector showed the least amount of substrate utilization (Supplementary Table 3). Molecular profiling Molecular characterization of actinomyces from different sectors of Chilika lake was carried out based on 16S rRNA gene sequencing and the results revealed that, ten isolates showed 100 % sequence identity compared with the most closely related sequences in a public database (Micromonospora echinospora, Streptomyces albogriseolus, S. bacillaris, S. achromogenes, S. fradiae, S. macrosporeus, S. ghanaensis, S. aureofaciens, S. erythrogriseus, S. fumigatiscleroticus), followed by nine isolates
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Table 2 Phenotypic characterization and closest BLASTN matches for the full 16S rDNA sequences and their percentage similarity with the closest alkali-halophilic actinobacterial strains Strain code
GenBank accession number
Closest GenBank match (% identity)
Colour of aerial mycelium
Colour of substrate mycelium
Diffusible pigment
Type of spore chain morphology and/or surface ornamentation
S1CS1
JN400094
None
Brown red
Orange
Spiny spore
S2NW2
JN400095
Grey
Yellow brown
None
Rectiflexibilis, smooth
S2NS5 S3RW3 S3RS5 S3RS8
JN400096 JN400097 JN400098 JN400099
Green gray White Grey Grey
Colourless Yellow Brown Black grey
None None None None
Retinaculiaperti, hairy Rectiflexibilis Spira Retinaculiaperti
S3RS9 S4BW2 S4BS3 S4BS5
JN885186 JN400100 JN400101 JN400102
Micromonospora echinospora (100 %) Streptomyces albogriseolus (100 %) Streptomyces acrimycini (99 %) Streptomyces albus (98 %) Streptomyces mutabilis (99 %) Streptomyces thermocarboxydus (99 %) Streptomyces atrovirens(99 %) Streptomyces bacillaris (100 %) Streptomyces geysiriensis (98 %) Streptomyces achromogenes (100 %)
None Yellow Grey Cream
Sand yellow Brown Yellow Yellow
Brown Brown None Brown
– Smooth Spira, hairy Rectiflexibilis
S4BS6 S5MW2 S5MS1 S5MS2 S5MS3 S5MS6 S5MS7 S6SW1 S6SS1 S6SS2 S6SS3
JN885189 JN400103 JN400104 JN400105 JN400106 JN400107 JN885187 JN400108 JN400109 JN400110 JN400111
None Grey Pink Grey Grey White Green Grey Whitish pink Greenish grey Pinkish white
Pastel orange Yellow Colourless Colourless Lemon yellow Yellow Green Colourless Beige Brown Yellow brown
None None None None None Brown None None None Brown None
Smooth Smooth, straight Retinaculiaperti, smooth Rectiflexibilis, smooth Spira Retinaculiaperti Spira, hairy Rectiflexibilis, smooth Rectiflexibilis, smooth Smooth, spira Smooth, spira
Micromonospora rosaria (99 %) Streptomyces vinaceusdrappus (99 %) Streptomyces fradiae (100 %) Streptomyces macrosporeus (100 %) Streptomyces griseorubens (99 %) Streptomyces labedae (99 %) Streptomyces ghanaensis(100 %) Streptomyces aureofaciens (100 %) Streptomyces spiralis (99 %) Streptomyces erythrogriseus (100 %) Streptomyces fumigatiscleroticus (100 %)
with 99 % similarity (Micromonospora rosaria, Streptomyces acrimycini, S. mutabilis, S. thermocarboxydus, S. atrovirens, S. vinaceusdrappus, S. griseorubens, S. labedae, S. spiralis) and lastly two isolates showing 98 % similarity (S. albus and S. geysiriensis) (Table 2; Figure 6).
Discussion Aquatic ecosystems have a strong functional linkage as producers and decomposers due to high substrate richness as well as functional diversity causing high heterogeneity in environmental resources. Chilika is a well explored lake in respect to physio-chemical, micro/macroscopic flora and fauna with an assemblage of marine (Sea mouth shore), brackish (Manika patana) and freshwater eco-systems (South and Central sectors) which supports a diverse and dynamic assemblage of microflora/fauna (Rath and Adhikary 2005). Isolation of culturable actinobacteria does not need an absolute requirement of sea water for growth (Maldonado et al. 2005). Isolation of actinomycetes results found that,
marine sectors (Manika patana and Sea mouth), showed greater species richness as compared to South and Central sectors, which may be due to formation of a nutrient gradient formed by tidal and fresh water from delta estuaries. Earlier workers also reported that nutrient availability, salinity and tidal factors are the limiting factors, which determine the distribution and ecology of microbes (Schafer et al. 2001). Study results revealed that, S. griseorubens, S. fradiae, M. echinospora, M. rosaria, S. mutabilis and S. albogriseolus are of aquatic origin which supports data of earlier workers (Mincer et al. 2002; Pathom-aree et al. 2006; Yoshida et al. 2008; Li 2009; Ye et al. 2009). Characterization of isolates is not only important to understand the role of these microbes in the particular niche but also to understand the requirement and conditions for their survivability in the particular specific niches (Ramesh and Mathivanan 2009). Characterization of isolates revealed that, south sector, central sector and sea mouth sectors had a maximum percentage of siderophore producers, followed by IAA and protease enzyme producers and nitrate reductase activity, respectively. Another observation revealed that
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Fig. 2 Scanning Electron Micrographs of Streptomyces strains isolated from Chilika lake, India, showing the different spore chain morphology of: a S3RS5 b S3RS8 c S3RW3 d S4BS3 e S4BW2 f S5MS3 g S6SS1 h S5MS2 i S6SS2
chitinase and protease enzyme producers were procured from sediment samples, while cellular siderophore, IAA, antimicrobial activity and nitrate reductase potential activity was highest in water samples. Actinomycetes can aid in plant
Fig. 3 Distribution and population frequencies of alkalihalophilic actinomycetes from different sectors of Chilika lake
growth promotion by IAA production for root growth or produce siderophores to improve nutrient uptake (Merckx et al. 1987) as well as having the siderophores also promote auxin synthesis by chelating metals (Dimkpa et al. 2008).
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Fig. 4 Profiling of 59 “morphotypes” of actinomycetes isolated from water and sediment of different sectors of Chilika lake for plant growth promotion and hydrolytic enzymes
Chitinolytic alkali-halophilic actinomycetes were more pronounced at the Sea mouth and Manika patana sectors due to presence of chitin rich substrates at the Sea mouth sector being fed by tidal waves from the Bay of Bengal as compared to other sectors. It is well known that Streptomyces species are capable of inhibiting pathogens by producing extracellular cell wall degrading enzymes (Thangapandian et al. 2007; Houssam 2009). While many actinomycetes show a relatively broad spectrum of biological activity, it is important to emphasise that a broad spectrum of activity may be due to multiple compound Fig. 5 Scanning Electron Micrograph of S. vinaceusdrappus strain S5MW2 and R. solani interaction after five days. (a) Healthy mycelium, control, (b) formation of spore chains on the fungal hyphae, (c) spore development inside the hyphae, (d) complete destruction of the fungal hypha
secreted by actinomycetes rather than a single inhibitory compound. In this study, we reported, S. vinaceusdrappus strain S5MW2 from the marine sector of Chilika (first time from India), showing strong antifungal properties against all test phytopathogens reported earlier (Mitra et al. 2010). The mode of action of S. vinaceusdrappus strain S5MW2 on phytopathogens analyzed by SEM showed gradual destruction of hyphae leading to death due to cytoplasmic extrusion, and it once again validates their potency to inhibit pathogens, although the exact mode of action of this strain is under investigation.
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Fig. 6 Phylogenetic tree based on the 16S rRNA gene sequences of alkali-halophilic actinomycetes isolates and their closest phylogenetic relatives. The tree was created by the neighbor-joining method. The boot
strap values from 5,000 pseudo-replications are shown at each of the branch points on the tree. Bar indicates % similarity
Many species of Streptomyces were reported to grow over a restricted pH range with pH optima of 7.0, but little attention has been given to alkali-halotolerant actinomycetes (Babu and Goodfellow 2008; Hozzein et al. 2011). The study also indicated that, species richness and alkalihalophilic actinomycetes population showed inverse
relationships with each other. We found out that the alkali-halophilic actinomycetes population was highest on marine sectors as compared to South and Central sectors of Chilika lake. This distribution might be due to specific epitopes across marine habitat rather than fresh water (Central and South) sectors.
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Identification through classical approaches were used to clarify the relationship within the genus Streptomyces, but since the last three decades, after intervention of 16S rRNA gene based sequences and phylogenetic analysis, many Streptomyces species can now be divided into multimembered species groups (Kumar and Goodfellow 2008). BLAST analysis of 16S rDNA gene sequences generated by sequencing of alkali-halophilic actinomycetes isolates revealed that two isolates had 98 % similarity with S. geysiriensis and S. albus while nine and ten isolates showed 99 % and 100 % similarity with other Streptomyces species, with two exceptions, which belong to the genus Micromonospora (M. echinospora, M. rosaria) (Table 2). Phylogenetic analysis representing three phyloclades with S. albus, S. bacillaris, S. fradiae S. achromogenes, S. fumigatiscleroticus S. spiralis falls under the first group while, S. albogriseolus, S. acrimycini, S. mutabilis, S. thermocarboxydus, S. geysiriensis, S. vinaceusdrappus, S. macrosporeus, S. griseorubens, S. labedae, S. aureofaciens, S. erythrogriseus, S. atrovirens, and S. ghanaensis represent a second group and lastly M. rosaria, M. echinospora form a third group showing a complete out group among each other. Isolates S3RS8, S4BS3 and S3RW3 seems to be novel as they showed similarity with many Streptomyces species but polyphasic characterization including morphology and physiological characteristics showed similarity with S. thermocarboxydus, S. geysiriensis and S. albus, respectively. Further, these isolates need to be validated for identification by DNA–DNA hybridization (%) GC content and FAME analysis up to species level, which is under progress.
Conclusion The actinomycetes population in different sectors of Chilika lake was considerably varied with variation in physiological as well as biochemical profiles which enabled us to understand behavioural, ecological, as well as specific substrate requirements in a particular brackish niche. The present findings not only showed the presence of alkali-halophilic streptomycetes in a halophilic and marine habitat but also provide an invaluable source for their taxonomic variations, distribution and role of these microbes in a particular niche as well as they are a rich source of bioactive compounds. Among them, S. vinaceusdrappus strain S5MW2, proved to have a potency to inhibit pathogens, however characterization of its mode of action is yet to be explored. The marine environment is a good source of an alkali-halophilic Streptomyces population having potentially new bioactive compounds which can be important in future bioprospecting in agricultural, industrial and pharmacological sectors. Further, emphasis should be made on research based on spatial and seasonal fluctuation of the actinomycetes population as well as employing new
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methodologies for the isolation and diversity analysis of rare actinomycetes genera. Acknowledgments The study was supported by a grant from the Indian Council of Agricultural Research (ICAR) New Delhi, India, under the Network Project ‘Application of Microorganisms in Agriculture and Allied Sectors (AMAAS)’.
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