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Isolation and characterization of marine bacteria from East Coast of India: functional screening for salt stress tolerance.

ABSTRACT: Soil salinization has become a severe constraint for crop production world-wide which necessitated development or induced enhancement of salt stress tolerance in plant life to sustain production in saline lands. Recognition and prospecting of valuable stress tolerant genes from natural microbial resources of saline habitat is obscure to date. Therefore, the investigation was towards isolation and characterization of marine salt stress tolerant microbes along the East coast of India for revelation of effective salt stress tolerant genes. Salt stress tolerance was assessed from 98 bacterial isolates obtained from 28 water and soil samples. Among them, 35 isolates which failed to grow beyond 4% salt were discarded and remainder 63 isolates were selected for further functional analysis and only seven isolates recorded ?8% NaCl stress tolerance. Phylogeny revealed that four isolates belong to Firmicutes and three isolates were members of Proteobacteria. Ribosomal Database Project Release-11 and SILVA SSU database based genotyping and taxonomic identity analysis confirmed that the higher (20%) salt stress tolerant bacteria were Staphylococcus sp., Enterococcus sp., Enterobacter sp. and Proteus sp. To investigate candidate, as well as, novel salt stress tolerant genes, the seven bacterial isolates would provide new horizon to focus on the recent developments of salinity stress tolerance. In addition, the findings evidently point out the diversity of salt stress tolerant marine bacteria in coastal Odisha and West Bengal, India.

SUBMITTER: Das P

PROVIDER: S-EPMC6581878 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Isolation and characterization of marine bacteria from East Coast of India: functional screening for salt stress tolerance.

Das Priyanka P Chatterjee Soumendranath S Behera Bijay Kumar BK Dangar Tushar Kanti TK Das Basanta Kumar BK Mohapatra Trilochan T

Heliyon 20190613 6

Soil salinization has become a severe constraint for crop production world-wide which necessitated development or induced enhancement of salt stress tolerance in plant life to sustain production in saline lands. Recognition and prospecting of valuable stress tolerant genes from natural microbial resources of saline habitat is obscure to date. Therefore, the investigation was towards isolation and characterization of marine salt stress tolerant microbes along the East coast of India for revelatio ...[more]

PMID: 31245639

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Project description:In the present study, we identified salt stress tolerant genes from the marine bacterium Staphylococcus sp. strain P-TSB-70 through transcriptome sequencing. In favour of whole-genome transcriptome profiling of Staphylococcus sp. strain P-TSB-70 (GenBank Accn. No. KP117091) which tolerated upto 20% NaCl stress, the strain was cultured in the laboratory condition with 20% NaCl stress. Transcriptome analyses were performed by SOLiD4.0 sequencing technology from which 10280 and 9612 transcripts for control and treated, respectively, were obtained. The coverage per base (CPB) statistics were analyzed for both the samples. Gene ontology (GO) analysis has been categorized at varied graph levels based on three primary ontology studies viz. cellular components, biological processes, and molecular functions. The KEGG analysis of the assembled transcripts using KAAS showed presumed components of metabolic pathways which perhaps implicated in diverse metabolic pathways responsible for salt tolerance viz. glycolysis/gluconeogenesis, oxidative phosphorylation, glutathione metabolism, etc. further involving in salt tolerance. Overall, 90 salt stress tolerant genes were identified as of 186 salt-related transcripts. Several genes have been found executing normally in the TCA cycle pathway, integral membrane proteins, generation of the osmoprotectants, enzymatic pathway associated with salt tolerance. Recognized genes fit diverse groups of salt stress genes viz. abc transporter, betaine, sodium antiporter, sodium symporter, trehalose, ectoine, and choline, that belong to different families of genes involved in the pathway of salt stress. The control sample of the bacterium showed elevated high proportion of transcript contigs (29%) while upto 20% salt stress treated sample of the bacterium showed a higher percentage of transcript contigs (31.28%). A total of 1,288 and 1,133 transcript contigs were measured entirely as novel transcript contigs in both control and treated samples, respectively. The structure and function of 10 significant salt stress tolerant genes of Staphylococcus sp. have been analyzed in this study. The information acquired in the present study possibly used to recognize and clone the salt stress tolerant genes and support in developing the salt stress-tolerant plant varieties to expand the agricultural productivity in the saline system.

Project description:Water deficit stress is a major contributor to the loss of ornamental crop value due to its negative effects on plant growth and flowering. In addition, post-production water stress can reduce the photosynthetic capacity of plants, negatively impacting crop quality at retail and in the consumer's home and garden. While the application of microbe-containing biostimulant products can increase stress tolerance and crop quality, the success of most commercially available biostimulants in greenhouse production systems is inconsistent. To identify beneficial bacteria with consistent biostimulant activity in greenhouse ornamentals, our study isolated bacteria from the rhizosphere of water stressed greenhouse ornamentals. Five species of popular ornamentals were obtained from 15 different commercial greenhouse facilities and then subjected to three cycles of water stress. Over 1,000 bacterial isolates were collected from the rhizosphere and screened in vitro for osmoadaptability and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. Eighty selected isolates were evaluated in a high-throughput greenhouse trial for their ability to increase plant size and flower number of water-stressed Petunia × hybrida. Ten bacterial strains selected from the high-throughput trial were then evaluated in a greenhouse validation trial for their ability to increase plant growth and to mitigate the reduction in photosynthetic parameters of water stressed P. hybrida and Pelargonium × hortorum. Application of certain bacteria increased plant size in both species and flower number of P. hybrida after recovery from water stress when compared to untreated water stressed plants. In addition, bacteria application increased the chlorophyll fluorescence parameters including quantum yield and efficiency of photosystem II (PSII) and electron transport rate (ETR), while decreasing the extent of electrolyte leakage during water stress and recovery. Overall, this study identified bacterial strains that can increase tolerance to and recovery from water stress in two commercially important ornamental crop species.

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Project description:The phylogenetic diversity of cultivable actinobacteria isolated from sponges (Haliclona spp.) and associated intertidal zone environments along the northern parts of the western coast of India were studied using 16S rRNA gene sequences. A subset of randomly selected actinobacterial cultures were screened for three activities, namely predatory behaviour, antibacterial activity and enzyme inhibition. We recovered 237 isolates from the phylum Actinobacteria belonging to 19 families and 28 genera, which could be attributed to 95 putative species using maximum-likelihood partition and 100 putative species using Bayesian partition in Poisson tree processes. Although the trends in the discovery of actinobacterial genera isolated from sponges were consistent with previous studies from different study areas, we provide the first report of nine actinobacterial species from sponges. We observed widespread non-obligate epibiotic predatory behaviour in eight actinobacterial genera and we provide the first report of predatory activity in Brevibacterium, Glutamicibacter, Micromonospora, Nocardiopsis, Rhodococcus and Rothia. Sponge-associated actinobacteria showed significantly more predatory behaviour than environmental isolates. While antibacterial activity by actinobacterial isolates mainly affected Gram-positive target bacteria with little or no effect on Gram-negative bacteria, predation targeted both Gram-positive and Gram-negative prey with equal propensity. Actinobacterial isolates from both sponges and associated environments produced inhibitors of serine proteases and angiotensin-converting enzyme. Predatory behaviour was strongly associated with inhibition of trypsin and chymotrypsin. Our study suggests that the sponges and associated environments of the western coast of India are rich in actinobacterial diversity, with widespread predatory activity, antibacterial activity and production of enzyme inhibitors. Understanding the diversity and associations among various actinobacterial activities – with each other and the source of isolation – can provide new insights into marine microbial ecology and provide opportunities to isolate novel therapeutic agents.

Project description:While marine microorganisms are frequently studied in their natural environment, isolated strains are invaluable resources that can be used in controlled experiments to expand upon direct observations from natural systems. Here, we sought a means to enhance culture collections of SAR11 marine bacteria by testing the use of seawater cryopreserved with glycerol as an inoculum. Using a raw seawater sample collected from the tropical Pacific Ocean, a subsample was diluted in seawater growth medium to create 576?2-ml dilution cultures containing 5 cells each and incubated for a high-throughput culturing (HTC) experiment, while another portion was cryopreserved in 10% glycerol. After 10 months, a cryopreserved aliquot was thawed and used to create a second cultivation experiment of 480?2-ml cultures containing 5 cells each and 470 cultures containing 105 cells each. The raw seawater cultivation experiment resulted in the successful isolation of 54 monocultures and 29 mixed cultures, while cryopreserved seawater resulted in 59 monocultures and 29 mixed cultures. Combined, the cultures included 51 SAR11 isolates spanning 11 unique 16S rRNA gene amplicon sequence variants (ASVs) from the raw seawater inoculum and 74 SAR11 isolates spanning 13 unique ASVs from cryopreserved seawater. A vast majority (92%) of SAR11 isolates from the two HTC experiments were members of SAR11 subclade Ia, though subclades IIIa and Va were also recovered from cryopreserved seawater and subclade Ib was recovered from both. The four most abundant SAR11 subclade Ia ASVs found in the initial seawater environmental sample were isolated by both approaches.IMPORTANCE High-throughput dilution culture has proved to be a successful approach to bring some difficult-to-isolate planktonic microorganisms into culture, including the highly abundant SAR11 lineage of marine bacteria. While the long-term preservation of bacterial isolates by freezing them in the presence of cryoprotectants, such as glycerol, has been shown to be an effective method of storing viable cells over long time periods (i.e., years), to our knowledge it had not previously been tested for its efficacy in preserving raw seawater for later use as an inoculum for high-throughput cultivation experiments. We found that SAR11 and other abundant marine bacteria could be isolated from seawater that was previously cryopreserved for nearly 10?months at a rate of culturability similar to that of the same seawater used fresh, immediately after collection. Our findings (i) expand the potential of high-throughput cultivation experiments to include testing when immediate isolation experiments are impractical, (ii) allow for targeted isolation experiments from specific samples based on analyses such as microbial community structure, and (iii) enable cultivation experiments across a wide range of other conditions that would benefit from having source inocula available over extended periods of time.

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Isolation and characterization of marine bacteria from East Coast of India: functional screening for salt stress tolerance.

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Isolation and characterization of marine bacteria from East Coast of India: functional screening for salt stress tolerance.

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