Introgression of a novel cold and drought regulatory-protein encoding CORA-like gene, SbCDR, induced osmotic tolerance in transgenic tobacco.

A potent cold and drought regulatory-protein encoding gene, SbCDR was cloned from an extreme halophyte Salicornia brachiata. In vitro localisation study, performed with SbCDR::RFP gene-construct revealed that SbCDR is a membrane protein. Overexpression of the SbCDR gene in tobacco plants confirmed tolerance against major environmental constraints such as salinity, drought and cold, as evidenced by improved chlorophyll contents, plant morphology, plant biomass, root length, shoot length and seed germination efficiency. Transgenic lines also exhibited high accumulation of proline, total sugar, reducing sugar, free amino acid and polyphenol, besides the low level of malondialdehyde (MDA) contents. SbCDR transgenic lines showed better relative water contents, membrane stability index and osmotic water potential. Furthermore, higher expression of ROS scavenging genes was observed in transgenic lines under stress. Moreover, microarray analysis revealed that several host genes were upregulated and downregulated under drought and salt stress conditions in SbCDR transgenic line compared with control (WT) plants. The results demonstrated that the overexpression of the halophytic SbCDR gene has intense effects on the abiotic stress tolerance of transgenic tobacco plants. However, the exact mode of action of SbCDR in multiple abiotic stress tolerance of plants is yet to be unveiled. It is believed that the precise role of SbCDR gene will provide additional information to comprehend the abiotic stress tolerance mechanism. Furthermore, it will appear as a promising candidate gene for improving stress tolerance in different crop plants for sustainable agriculture and crop productivity.

Marine bacterial biodegradation of low-density polyethylene (LDPE) plastic.

Polyethylene has considered as non-degradable for decades, and their degradation through marine bacteria has rarely studied. However, LDPE found a significant source of pollution in the marine environment. In the present study, four bacterial strains capable of biodegradation of LDPE were isolated from the marine environment. These bacterial isolates H-237, H-255, H-256 and H-265 were revealed close similarity with Cobetia sp., Halomonas sp., Exigobacterium sp. and Alcanivorax sp., respectively based on 16S rRNA gene sequencing method. These bacterial isolates were individually incubated for 90 days supplied with LDPE films as a carbon source using the Bushnell-Haas medium. During the biodegradation assay, bacterial isolates were formed the viable biofilm on the LDPE surface, which decreased the thermal stability of the films. At the end of the incubation study, a maximum weight loss of 1.72% of LDPE film was observed by the bacterial isolate H-255. The bacterial attachment on the film changed the physical structure (surface erosion, roughness and degradation) which were confirmed by field emission scanning electron microscopy and atomic force microscopy. The changes in the chemical structure of the LDPE film were analyzed by Attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR). This ATR-FTIR showed the shifting of peaks of C-H stretch and C=C bond stretching and the new peaks formation of C-O and -C=C- bonds in comparison to control LDPE film. Further, biodegradation of LDPE film was also confirmed by remineralization of carbon and enzymatic activities. This study revealed that the active biodegradation of LDPE film by marine bacteria and these bacteria could reduce plastic pollution in the marine environment.

Draft genome sequence of plastic degrading Bacillus sp. AIIW2 isolated from the Arabian ocean.

The endemic spread of plastic in the environment requires urgent need of a sustainable approach. Marine microbes found to have vast bioactivity and play a central role in biogeochemical cycling in the ocean; however, very few of them had been explored for biochemical cycling or plastic degradation. In the present study, we report the draft genome sequence of marine Bacillus sp. AIIW2 which was found to utilize plastic as a carbon source. The Bacillus sonorensis SRCM101395 was used as a reference genome for mapping the reads. The genome size of strain AIIW2 was approximately 4.4 Mb and composed of 4737 coding sequences with 45.7% G + C contents. The whole genome comparison of strain AIIW2 with three closest Bacillus strains showed strain specificity, the 16S ribosomal RNA sequence shows 99.93% similarity with Bacillus paralicheniformis KJ-16T (KY694465). This genome data would provide the genetic basis in developing plastic bioremediation approaches and discover the enzymes pertinent in the biodegradation processes.

Metabolic profiling and scavenging activities of developing circumscissile fruit of psyllium (Plantago ovata Forssk.) reveal variation in primary and secondary metabolites.

BACKGROUND: Developing fruit is considered as an excellent model to study the complex network of metabolites which are altered rapidly during development. RESULTS: Metabolomics revealed that developing psyllium fruit is a rich source of primary metabolites (ω-3 and ω-6 fatty acids and amino-acids), secondary metabolites and natural antioxidants. Eidonomy and anatomy confirmed that psyllium fruit followed five stages of development. Total lipids and fatty acids were synthesized differentially; saturated fatty acids (FAs) increased, whereas total polyunsaturated FAs decreased with increasing developmental stage. The unsaturation index and degree of unsaturation showed a catenary curve. Principal component analysis confirmed a significant shift in the FA profile from bud initiation to the maturation stage. Similarly, a similar level of total amino acids was present at different developmental stage following a temporal biosynthesis pathway. Total phenolic and flavonoid contents decreased in tandem with fruit development. Twenty-two different metabolites were identified, and metabolic changes were also observed during fruit development. Six metabolites were detected exclusively in the flowering stage, whereas two were detected in each of early and maturity stages of development. The metabolites apigenin and kaempferol were detected ubiquitously in all developmental stages. Time-dependent metabolomics revealed a shift in metabolite biosynthesis. CONCLUSION: During fruit development, metabolites, FAs, amino acids, total phenolics, total flavonoids, antioxidants and scavenging activities changed progressively and were co-ordinately linked to each other. As a future perspective, further studies will focus on the validation of identified metabolites, which integrated with transcriptomics data and will reveal the metabolic regulatory network of development psyllium fruit.

Introgression of UfCyt c6, a thylakoid lumen protein from a green seaweed Ulva fasciata Delile enhanced photosynthesis and growth in tobacco.

Cytochromes are important components of photosynthetic electron transport chain. Here we report on genetic transformation of Cytochrome c6 (UfCyt c6) gene from Ulva fasciata Delile in tobacco for enhanced photosynthesis and growth. UfCyt c6 cDNA had an open reading frame of 330 bp encoding a polypeptide of 109 amino acids with a predicted molecular mass of 11.65 kDa and an isoelectric point of 5.21. UfCyt c6 gene along with a tobacco petE transit peptide sequence under control of CaMV35S promoter was transformed in tobacco through Agrobacterium mediated genetic transformation. Transgenic tobacco grew normal and exhibited enhanced growth as compared to wild type (WT) and vector control (VC) tobacco. Transgenic tobacco had higher contents of photosynthetic pigments and better ratios of photosynthetic pigments. The tobacco expressing UfCyt c6 gene exhibited higher photosynthetic rate and improved water use efficiency. Further activity of the water-splitting complex, photosystem II quantum yield, photochemical quenching, electron transfer rate, and photosynthetic yield were found comparatively higher in transgenic tobacco as compared to WT and VC tobacco. Alternatively basal quantum yield of non-photochemical processes in PSII and non-photochemical quenching were estimated lower in tobacco expressing UfCyt c6 gene. As a result of improved photosynthetic performance the transgenic tobacco had higher contents of sugar and starch, and exhibited comparatively better growth. To the best of our knowledge this is the first report on expression of UfCyt c6 gene from U. fasciata for improved photosynthesis and growth in tobacco.

Spatial and halophyte-associated microbial communities in intertidal coastal region of India.

Microbial communities in intertidal coastal soils respond to a variety of environmental factors related to resources availability, habitat characteristics, and vegetation. These intertidal soils of India are dominated with Salicornia brachiata, Aeluropus lagopoides, and Suaeda maritima halophytes, which play a significant role in carbon sequestration, nutrient cycling, and improving microenvironment. However, the relative contribution of edaphic factors, halophytes, rhizosphere, and bulk sediments on microbial community composition is poorly understood in the intertidal sediments. Here, we sampled rhizosphere and bulk sediments of three dominant halophytes (Salicornia, Aeluropus, and Suaeda) from five geographical locations of intertidal region of Gujarat, India. Sediment microbial community structure was characterized using phospholipid fatty acid (PLFA) profiling. Microbial biomass was significantly influenced by the pH, electrical conductivity, organic carbon, nitrogen, and sodium and potassium concentrations. Multivariate analysis of PLFA profiles had significantly separated the sediment microbial community composition of regional sampling sites, halophytes, rhizosphere, and bulk sediments. Sediments from Suaeda plants were characterized by higher abundance of PLFA biomarkers of Gram-negative, total bacteria, and actinomycetes than other halophytes. Significantly highest abundance of Gram-positive and fungal PLFAs was observed in sediments of Aeluropus and Salicornia, respectively than in those of Suaeda. The rhizospheric sediment had significantly higher abundance of Gram-negative and fungal PLFAs biomarkers compared to bulk sediment. The results of the present study contribute to our understanding of the relative importance of different edaphic and spatial factors and halophyte vegetation on sediment microbial community of intertidal sediments of coastal ecosystem.

Biomass production, nutrient cycling, and carbon fixation by Salicornia brachiata Roxb.: A promising halophyte for coastal saline soil rehabilitation.

In order to increase our understanding of the interaction of soil-halophyte (Salicornia brachiata) relations and phytoremediation, we investigated the aboveground biomass, carbon fixation, and nutrient composition (N, P, K, Na, Ca, and Mg) of S. brachiata using six sampling sites with varying characteristics over one growing season in intertidal marshes. Simultaneously, soil characteristics and nutrient concentrations were also estimated. There was a significant variation in soil characteristics and nutrient contents spatially (except pH) as well as temporally. Nutrient contents in aboveground biomass of S. brachiata were also significantly differed spatially (except C and Cl) as well as temporally. Aboveground biomass of S. brachiata ranged from 2.51 to 6.07 t/ha at maturity and it was positively correlated with soil electrical conductivity and available Na, whereas negatively with soil pH. The K/Na ratio in plant was below one, showing tolerance to salinity. The aboveground C fixation values ranged from 0.77 to 1.93 C t/ha at all six sampling sites. This study provides new understandings into nutrient cycling-C fixation potential of highly salt-tolerant halophyte S. brachiata growing on intertidal soils of India. S. brachiata have a potential for amelioration of the salinity due to higher Na bioaccumulation factor.

Methyl Jasmonate-Induced Lipidomic and Biochemical Alterations in the Intertidal Macroalga Gracilaria dura (Gracilariaceae, Rhodophyta).

The role of exogenously added methyl jasmonate (MeJA), a lipid-derived signaling compound, in inducing oxidative stress in the marine red macroalga Gracilaria dura was investigated. MeJA at a concentration of 1-100 µM was a strong stimulant of reactive oxygen species (H(2)O(2), HO· and O(2) (·-)) (P < 0.05) causing considerable oxidative stress in G. dura. This further led to lipid peroxidation and degradation of the pigments Chl a and phycocyanin, with a concomitant increase in phycoerythrin. The MeJA-induced oxidative burst also led to the induction of a fatty acid oxidation cascade, resulting in the synthesis of hydroxy-oxylipins and the up-regulation of the 13-lipoxygenase pathway. Electrospray ionization-mass spectrometry-based shotgun lipidomic analysis revealed that monogalactosyldiacylglycerol (a chloroplastic glycerolipid) and phosphatidylcholine (extrachloroplastidic phopholipid) were the most affected lipid classes. The degradation of 18:3-fatty acid-containing monogalactosyldiacylglycerol inferred that it provided fatty acyl chains for the biosynthesis of 13-hydroperoxylinolenic acid, which was further directed towards either the jasmonate pathway or other alternative pathways of the fatty acid oxidation cascade, analogous to higher plants. Also, G. dura modulated the lipid acyl chains in such a way that no significant change was observed in the fatty acid profile of the treated thalli as compared with those of the control, except for C16:0, C16:1 (n-9), C20:3 (n-6) and C20:4 (n-6) (P < 0.05). Furthermore, MeJA caused the accumulation of phenolic compounds and the up-regulation of enzymes involved in secondary metabolism such as polyphenol oxidase, shikimate dehydrogenase and phenylalanine ammonia-lyase, indicating a shift towards secondary metabolism as a defense strategy to combat the induced oxidative stress.

Overexpression of a novel SbMYB15 from Salicornia brachiata confers salinity and dehydration tolerance by reduced oxidative damage and improved photosynthesis in transgenic tobacco.

MAIN CONCLUSION: SbMYB15, R2R3-type MYB was induced by the different stresses, and conferred stress tolerance in transgenic tobacco by regulating the expression of stress-responsive genes. MYBs are the master regulators of various metabolic processes and stress responses in plants. In this study, we functionally characterised a R2R3-type SbMYB15 transcription factor (TF) from the extreme halophyte Salicornia brachiata. The SbMYB15 acts as a transcriptional activator. Transcriptional analysis showed that SbMYB15 transcript was strongly upregulated in red shoots and was also induced by different stresses; however, its expression remained unchanged with ABA. Overexpression of SbMYB15 in tobacco significantly improved salinity and dehydration tolerance. The enhanced tolerance of the transgenic plants was defined by the changes in chlorophyll, malondialdehyde (MDA), proline, total soluble sugar and total amino acid contents. The transgenic plants exhibited a higher membrane stability and reduced electrolyte leakage, H2O2 and O 2 (-) content compared to the wild type (WT). With ionic stress, transgenics showed a low Na(+) and a high K(+) content. In the transgenic plants, the expression of stress-responsive genes such as LEA5, ERD10D, PLC3, LTP1, HSF2, ADC, P5CS, SOD and CAT was enhanced in the presence of salinity, dehydration and heat. Exposure to gradual salinity and dehydration resulted in an increased stomatal conductance, water use efficiency, photosynthesis rate, photochemical quenching and reduced transpiration rate. Thus, SbMYB15 served as an important mediator of stress responses regulating different stress signalling pathways, leading to enhanced stress tolerance.

Zhihengliuella somnathii sp. nov., a halotolerant actinobacterium from the rhizosphere of a halophyte Salicornia brachiata.

Two novel, Gram-stain-positive, rod-shaped, halotolerent bacteria, strains JG 03(T) and JG 05 were isolated from the rhizosphere of Salicornia brachiata, an extreme halophyte. Comparative analyses of 16S rRNA gene sequences showed that they were closely related to members of the genus Zhihengliuella, with sequence similarities of 96.9-99.1%. The sequence similarity of strains JG 03(T )and JG 05 with each other was 99.4%. DNA-DNA hybridization of JG 03(T) and JG 05 with other species of the genus Zhihengliuella with validly published names showed reassociation values of 19.8%-53.4% and a value of 91.4% between each other. The peptidoglycan type of both strains was A4α and MK-9 and MK-10 were the predominant menaquinones. The predominant fatty acid in JG 03(T) was anteiso-C15 : 0 and anteiso-C17 : 0. However, iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0 were the major fatty acids in strain JG 05. The DNA G+C content of strains JG 03(T) and JG 05 was 70.0 and 70.1 mol%, respectively. In nutrient broth medium both strains grew at NaCl concentrations of up to 15% (w/v). On the basis of chemotaxonomic characteristics and phylogenetic analyses, strains JG 03(T) and JG 05 should be affiliated to the genus Zhihengliuella. Strains JG 03(T) and JG 05 represent a novel species of the genus Zhihengliuella for which the name Zhihengliuella somnathii sp. nov. is proposed. The type strain is JG 03(T) ( = DSM 23187(T) = IMCC 253(T)).

Nitrate and phosphate regimes induced lipidomic and biochemical changes in the intertidal macroalga Ulva lactuca (Ulvophyceae, Chlorophyta).

This study was carried out in order to understand the lipid and biochemical alterations resulting from different nutritional regimes of nitrate and phosphate in Ulva lactuca. The algal thalli cultured in artificial seawater (ASW) showed higher levels of carbohydrates and non-polar lipids and increased phosphatase activities, accompanied by degradation of polar lipids, proteins and pigments. Further, higher levels of lipid hydroperoxides indicated reative oxygen species (ROS)-mediated non-enzymatic lipid peroxidation due to nutritional limitation-induced oxidative stress. Those thalli cultured in ASW supplemented with nitrate showed responses corresponding to nitrate addition, such as an increase in pigments, monogalactosyldiacylglycerols, polyunsaturated fatty acids and nitrate reductase. In addition, these thalli showed partial induction of phosphatases, low phospholipids, and high sulfolipid and 1,2-diacylglyceryl-3-O-4'-(N,N,N-trimethyl)-homoserine (DGTS) due to phosphate limitation. Similarly, algal thalli cultured in ASW supplemented with phosphate showed down-regulation of phosphatases, an increase in phospholipids due to availability of phosphate as well as a decrease in nitrate reductase, pigment, monogalactosyldiacylglycerols and polyunsaturated fatty acids due to nitrate limitation. On the other hand, algal thalli cultured in ASW supplemented with both nitrate and phosphate showed recovery of lost pigments and proteins, a high monogalactosyldiacylglycerol/digalactosyldiacylglycerol ratio, high unsaturation and high oxylipin levels (both C18 and C20). Further, the accumulation of indole-3-acetic acid in nutrient-limited thalli and of kinetin and kinetin riboside in nutrient-supplemented thalli indicated their antagonistic roles under nutrient stress. Thus, U. lactuca copes with nitrate and phosphate nutritional stress by altering the metabolic pathways involved in lipid biosynthesis including a shift in lipid classes, fatty acids, oxylipins and indole-3-acetic acid/kinetin cross-talk.

The transcriptional regulatory mechanism of the peroxisomal ascorbate peroxidase (pAPX) gene cloned from an extreme halophyte, Salicornia brachiata.

Peroxisomal ascorbate peroxidase detoxifies H2O2 leaching out from peroxisomes into the cytoplasm. The present study describes transcript expression and cis-regulation of the SbpAPX gene cloned from an extreme halophyte, Salicornia brachiata, in the steady state and under different stresses. About 2-fold elevated transcript expression was found in salt- and drought-treated shoots at 12 h compared with control, while 1.9-fold increased expression was observed under heat treatment. In roots, the transcript level was down-regulated at 2 h, thereafter increasing with the time of exposure and reaching a maximum at the control level. The SbpAPX promoter has characteristic cis-regulatory ABA-dependent abiotic stress-responsive elements. The full-length promoter (1,024 bp, PP1) and deletion constructs -838 (PP2), -697 (PP3), -433 (PP4) and -185 bp (PP5) were fused with the GUS (ß-glucuronidase) gene and transformed into tobacco for functional validation. Expression of GUS increased significantly in transgenic plants under stress. Quantitative expression analysis of GUS in T1 plants revealed that promoter PP5 is efficient for gene expression. In planta transient expression further suggested that the promoter PP5 contains efficient stress-inducible elements. A steep decline in GUS expression in PP3, and thereafter an elevated expression in PP4 and PP5, suggested the presence of a repressor element between -696 and -433 bp, while an enhancer element was predicted between -838 and -697 bp. Further, transient expression analyses and electrophoretic mobility shift assay revealed that the core sequence of cis-acting motifs ATAA and CCTCAA function as enhancer and repressor binding sites, respectively. Based on the study, a model is proposed for the cis-regulation of the SbpAPX gene. The present study provides a useful insight for understanding gene expression regulation in a halophyte with or without stress. Furthermore, potential stress-responsive promoter-driven expression of introgressed gene(s) can be used for engineering crops with enhanced stress tolerance.

Cis-9-octadecenoic acid from the rhizospheric bacterium Stenotrophomonas maltophilia BJ01 shows quorum quenching and anti-biofilm activities.

Quorum quenching (QQ) is an effective approach for the prevention of bacterial infections involving biofilms. This study reports the QQ and anti-biofilm activities of a rhizospheric bacterium identified as Stenotrophomonas maltophilia BJ01. The QQ activity was demonstrated using Chromobacterium violaceum CV026 as a biosensor. A maximum of 95% reduction in violacein production, a quorum sensing-regulated behavior, was observed. Gas chromatography-mass spectroscopy of the extract showed that the active compound was cis-9-octadecenoic acid, which was confirmed by electronspray ionization-mass spectroscopy data. The extract also inhibited biofilm formation of Pseudomonas aeruginosa ATCC 9027 without affecting its growth. Scanning electron and atomic force microscopy showed architectural disruption of the biofilm when treated with the extract. This is the first report of the QQ and anti-biofilm activities of cis-9-octadecenoic acid isolated from any bacterium. It may have the potential to combat detrimental infections with P. aeruginosa. Further validation is required for any possible medical application.

Quorum sensing inhibition by Asparagopsis taxiformis, a marine macro alga: separation of the compound that interrupts bacterial communication.

The majority of the marine algal species, though completing their life cycle in seawater, are rarely susceptible to fouling, making them an important source of quorum sensing (QS) inhibitory substances. The separation and characterization of QS inhibitors are crucial for any potential application. Thirty marine macroalgae were tested for QS inhibition activity by using Chromobacterium violaceum CV026 as the reporter strain, and among them, Asparagopsis taxiformis showed antibacterial, as well as antiquorum, sensing activities. Cinnamaldehyde (75 mM) and methanol were used as positive and negative controls, respectively. The antiquorum sensing activity of A. taxiformis was further confirmed using the sensor strain, Serratia liquefaciens MG44, having green fluorescent protein (gfp). Methanolic extract of the alga was fractionated by solid phase extraction (SPE), and each fraction was tested for QS inhibition. Two types of activities were observed-zone of clearance (antibacterial activity) and zone of inhibition with or without finger-like projections (QS inhibition). Out of five SPE cartridges, Bond Elut PH showed clear separation of these two fractions. The Ion Cyclotron Resonance Fourier Transformation Mass Spectrometer (ICR-FT/MS) analysis of the fractions further supported the bioassay results. The presence of strong QS inhibitory compound in A. taxiformis indicates its potential use in antifouling preparations.

Surface-associated bacterial assemblages on marine anthropogenic litter in the intertidal zone of the Arabian Sea, India.

Anthropogenic marine litter (mainly plastic pollution) is a serious concern globally. The interactions between terrestrial and marine ecosystems lead to the accumulation of marine litter in the intertidal zone. The biofilm-forming bacteria tend to colonize on surfaces of marine litter which are composed of diverse bacteria and are less studied. The present study investigated the bacterial community composition using both culturable and non-culturable (Next-generation sequencing (NGS)) approaches associated with the marine litter (polyethylene (PE), styrofoam (SF) and fabric (FB)) at three distinct locations (Alang, Diu and Sikka) of the Arabian Sea, Gujarat, India. Predominant bacteria observed using culturable and NGS techniques belonged to Proteobacteria phyla. Alphaproteobacteria class dominated on polyethylene and styrofoam surfaces in the culturable fraction among the sites while the Bacillus dominated fabric surfaces. In the metagenomics fraction, Gammaproteobacteria dominated the surfaces except for PE and SF surfaces from Sikka and Diu, respectively. The PE surface at Sikka was dominated by Fusobacteriia while SF surface from Diu was dominated by Alphaproteobacteria. Both culture-dependent and NGS approaches identified hydrocarbon-degrading bacteria as well as pathogenic bacteria on the surfaces. The outcome of the present study illustrates diverse bacterial assemblages which occur on marine litter and increases our understanding of the plastisphere community.

The SbSOS1 gene from the extreme halophyte Salicornia brachiata enhances Na(+) loading in xylem and confers salt tolerance in transgenic tobacco.

BACKGROUND: Soil salinity adversely affects plant growth and development and disturbs intracellular ion homeostasis resulting cellular toxicity. The Salt Overly Sensitive 1 (SOS1) gene encodes a plasma membrane Na(+)/H(+) antiporter that plays an important role in imparting salt stress tolerance to plants. Here, we report the cloning and characterisation of the SbSOS1 gene from Salicornia brachiata, an extreme halophyte. RESULTS: The SbSOS1 gene is 3774 bp long and encodes a protein of 1159 amino acids. SbSOS1 exhibited a greater level of constitutive expression in roots than in shoots and was further increased by salt stress. Overexpressing the S. brachiata SbSOS1 gene in tobacco conferred high salt tolerance, promoted seed germination and increased root length, shoot length, leaf area, fresh weight, dry weight, relative water content (RWC), chlorophyll, K(+)/Na(+) ratio, membrane stability index, soluble sugar, proline and amino acid content relative to wild type (WT) plants. Transgenic plants exhibited reductions in electrolyte leakage, reactive oxygen species (ROS) and MDA content in response to salt stress, which probably occurred because of reduced cytosolic Na(+) content and oxidative damage. At higher salt stress, transgenic tobacco plants exhibited reduced Na(+) content in root and leaf and higher concentrations in stem and xylem sap relative to WT, which suggests a role of SbSOS1 in Na(+) loading to xylem from root and leaf tissues. Transgenic lines also showed increased K(+) and Ca(2+) content in root tissue compared to WT, which reflect that SbSOS1 indirectly affects the other transporters activity. CONCLUSIONS: Overexpression of SbSOS1 in tobacco conferred a high degree of salt tolerance, enhanced plant growth and altered physiological and biochemical parameters in response to salt stress. In addition to Na(+) efflux outside the plasma membrane, SbSOS1 also helps to maintain variable Na(+) content in different organs and also affect the other transporters activity indirectly. These results broaden the role of SbSOS1 in planta and suggest that this gene could be used to develop salt-tolerant transgenic crops.

Comparative molecular analysis of chemolithoautotrophic bacterial diversity and community structure from coastal saline soils, Gujarat, India.

BACKGROUND: Soils harbour high diversity of obligate as well as facultative chemolithoautotrophic bacteria that contribute significantly to CO2 dynamics in soil. In this study, we used culture dependent and independent methods to assess the community structure and diversity of chemolithoautotrophs in agricultural and coastal barren saline soils (low and high salinity). We studied the composition and distribution of chemolithoautotrophs by means of functional marker gene cbbL encoding large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and a phylogenetic marker 16S rRNA gene. The cbbL form IA and IC genes associated with carbon fixation were analyzed to gain insight into metabolic potential of chemolithoautotrophs in three soil types of coastal ecosystems which had a very different salt load and sulphur content. RESULTS: In cbbL libraries, the cbbL form IA was retrieved only from high saline soil whereas form IC was found in all three soil types. The form IC cbbL was also amplified from bacterial isolates obtained from all soil types. A number of novel monophyletic lineages affiliated with form IA and IC phylogenetic trees were found. These were distantly related to the known cbbL sequences from agroecosystem, volcanic ashes and marine environments. In 16S rRNA clone libraries, the agricultural soil was dominated by chemolithoautotrophs (Betaproteobacteria) whereas photoautotrophic Chloroflexi and sulphide oxidizers dominated saline ecosystems. Environmental specificity was apparently visible at both higher taxonomic levels (phylum) and lower taxonomic levels (genus and species). The differentiation in community structure and diversity in three soil ecosystems was supported by LIBSHUFF (P = 0.001) and UniFrac. CONCLUSION: This study may provide fundamentally new insights into the role of chemolithoautotrophic and photoautotrophic bacterial diversity in biochemical carbon cycling in barren saline soils. The bacterial communities varied greatly among the three sites, probably because of differences in salinity, carbon and sulphur contents. The cbbL form IA-containing sulphide-oxidizing chemolithotrophs were found only in high saline soil clone library, thus giving the indication of sulphide availability in this soil ecosystem. This is the first comparative study of the community structure and diversity of chemolithoautotrophic bacteria in coastal agricultural and saline barren soils using functional (cbbL) and phylogenetic (16S rDNA) marker genes.

A novel salt-inducible gene SbSI-1 from Salicornia brachiata confers salt and desiccation tolerance in E. coli.

Salicornia brachiata is one of the extreme salt tolerant plants and grows luxuriantly in coastal areas. Previously we have reported isolation and characterization of ESTs from S. brachiata with large number of unknown gene sequences. Reverse Northern analysis showed upregulation and downregulation of few unknown genes in response to salinity. Some of these unknown genes were made full length and their functional analysis is being tested. In this study, we have selected a novel unknown salt inducible gene SbSI-1 (Salicornia brachiata salt inducible-1) for the functional validation. The SbSI-1 (Gen-Bank accession number JF 965339) was made full length and characterized in detail for its functional validation under desiccation and salinity. The SbSI-1 gene is 917 bp long, and contained 437 bp 3' UTR, and 480 bp ORF region encoding 159 amino acids protein with estimated molecular mass of 18.39 kDa and pI 8.58. The real time PCR analysis revealed high transcript expression in salt, desiccation, cold and heat stresses. However, the maximum expression was obtained by desiccation. The ORF region of SbSI-1 was cloned in pET28a vector and transformed in BL21 (DE3) E. coli cells. The SbSI-1 recombinant E. coli cells showed tolerance to desiccation and salinity stress compared to only vector in the presence of stress.

An appraisal of early stage biofilm-forming bacterial community assemblage and diversity in the Arabian Sea, India.

The community composition and distribution of early-stage (24 h) biofilm-forming bacteria on two different surfaces (glass slide and polystyrene plastic slide) at three different locations (Diu, Alang and Sikka) were studied using a culture-dependent and next-generation sequencing (NGS) approach in the Arabian Sea, Gujarat, India. The most dominant phyla observed using the NGS approach were the Proteobacteria among the sampling sites. Gammaproteobacteria class dominated both the surfaces among the sites and accounted for 46.7% to 89.2% of total abundance. The culture-dependent analysis showed Proteobacteria and Firmicutes as the dominant phyla on the surfaces within the sampling sites. During the initial colonization, hydrocarbon-degrading bacterial strains have also attached to the surfaces. The outcome of this study would be of great importance for targeting the early stage biofilm-forming and hydrocarbon-degrading bacterial isolates may help to degrade plastic in the marine environment.

Marine bacteria-based polyvinyl chloride (PVC) degradation by-products: Toxicity analysis on Vigna radiata and edible seaweed Ulva lactuca.

Biodegradation of polyvinyl chloride (PVC) by marine bacteria is a sustainable approach that leads to the production of different by-products but their toxicity needs to be evaluated. In the present study, polyvinyl chloride degradation products (PVCDP) produced by three marine bacterial isolates (T-1.3, BP-4.3 and S-237) in the culture supernatant were evaluated for toxicity on the germination of Vigna radiata and growth of Ulva lactuca. A total of 24 compounds comprising of benzene, fatty acid, ether, ester and plastic stabilizer (tris (2, 4-di-tert-butylphenyl) phosphate) were identified by GC-MS using diethyl ether solvent extraction from the supernatant. The per cent germination rate of the seed treated with PVCDP showed no significant effect but germination index and elongation inhibition rate were influenced significantly by PVCDP treatments. In seaweed (U. lactuca), PVCDP showed improvement in the daily growth rate. After ten days of treatment with PVCDP, pigment contents were improved in seaweed and PVCDP (2%) of isolate T-1.3 recorded the highest chlorophyll-a and chlorophyll-b.