Hexagonal Boron Nitride Quantum Dots Embedded on Layer-by-Layer Films for Peroxidase-Assisted Colorimetric Detection of ß-Galactosidase Producing Pathogens.

Pathogen detection has become a major research area all over the world for water quality surveillance and microbial risk assessment. Therefore, designing simple and sensitive detection kits plays a key role in envisaging and evaluating the risk of disease outbreaks and providing quality healthcare settings. Herein, we have designed a facile and low-cost colorimetric sensing strategy for the selective and sensitive determination of ß-galactosidase producing pathogens. The hexagonal boron nitride quantum dots (h-BN QDs) were established as a nanozyme that showed prominent peroxidase-like activity, which catalyzes 3,3',5,5'-tetramethylbenzidine (TMB) oxidation by H2O2. The h-BN QDs were embedded on a layer-by-layer assembled agarose biopolymer. The ß-galactosidase enzyme partially degrades ß-1,4 glycosidic bonds of agarose polymer, resulting in accessibility of h-BN QDs on the solid surface. This assay can be conveniently conducted and analyzed by monitoring the blue color formation due to TMB oxidation within 30 min. The nanocomposite was stable for more than 90 days and was showing TMB oxidation after incubating it with Escherichia coli (E. coli). The limit of detection was calculated to be 1.8 × 106 and 1.5 × 106 CFU/mL for E. coli and Klebsiella pneumonia (K. pneumonia), respectively. Furthermore, this novel sensing approach is an attractive platform that was successfully applied to detect E. coli in spiked water samples and other food products with good accuracy, indicating its practical applicability for the detection of pathogens in real samples.

Impact of polyvinyl chloride (PVC) microplastic on growth, photosynthesis and nutrient uptake of Solanum lycopersicum L. (Tomato).

Microplastics (MPs) pollution and their impact on plants have become a global threat, but their effect at the molecular level remains scarce. This study aims to gain insight into the effects of polyvinylchloride microplastic (PVC-MP) on tomato plants at the genetic and protein levels. In this study, we found that increasing concentrations of PVC-MP (2.5, 5,7.5, and 10% w/w) in the soil did not cause any phytotoxic (chlorosis or necrosis) symptoms but it did result in a dose-dependent reduction in plant growth-related parameters, such as height, leaf area, stem diameter, and plant fresh and dry weight. Additionally, the number of secondary roots was reduced while the primary roots were elongated. Furthermore, PVC-MP also caused a significant decrease in light-harvesting pigments chlorophylls, and carotenoids while increasing the level of reactive oxygen species (ROS) and lipid peroxidation in plants. Microscopic analysis of the roots revealed the uptake of PVC-MP of size less than 10 µm. Micro- and macro-element analysis showed changes in concentrations of Ca, Cu, Fe, Mg, Mn, Ni, and Zn, upon PVC-MP exposure. Results from western blotting and q-PCR showed that higher doses of PVC-MP significantly reduced the CO2-fixing enzyme RuBisCO and D1 proteins of PSII at both protein and transcript levels. These findings suggest that lower levels of light-harvesting pigments, D1 protein, RuBisCO, and modulation of nutrient absorption are among the factors responsible for growth suppression in tomato plants upon exposure to PVC-MP. As tomato plants are economically significant crops, an increase in PVC-MP in agricultural fields may have a detrimental influence on crop production, resulting in economic loss.

Triple quadrupole liquid chromatography-mass spectrometry-mediated evaluation of vitamin D2 accumulation potential, antioxidant capacities, and total polyphenol content of white jelly mushroom (Tremella fuciformis Berk.).

Tremella fuciformis Berk. (TF), or the white jelly mushroom, is well known for its myriad of pharmacological properties, such as immunomodulatory, anti-inflammatory, antidiabetic, antitumor, and antioxidant activities, and hypocholesterolemic and hepatoprotective effects that boost human health. Most of the studies of TF are concentrated on its polysaccharide (glucuronoxylomannan) composition, which is responsible for its pharmacological as well as rheological properties. It is well established that mushrooms are a great source of dietary vitamin D due to the presence of ergosterol in their cell membrane. There is a lack of published data on TF as a source of vitamin D2. Therefore, this study aimed to evaluate the vitamin D2 composition of the fruiting bodies of TF using triple quadrupole liquid chromatography-mass spectrometry (LC-MS/QQQ). The results showed highest vitamin D2 content (292.02 µg/g dry weight) in the sample irradiated with ultraviolet B (UVB; 310 nm) for 180 min as compared with the control group (52.47 µg/g dry weight) (P ≤ 0.001). The results showed higher accumulation potential of vitamin D2 in TF as compared with published data available for other extensively studied culinary mushrooms, such as Agaricus bisporus, Lentinula edodes, Pleurotus ostreatus, Cordiceps militaris, and Calocybe indica. Moreover, the impact of UV treatment on antioxidant capacities and total polyphenol content of TF was also studied. The accumulation potential of vitamin D in TF reveals a novel commercial source for this nutrient.

Comparative seasonal analysis of Eri silkworm (Samia ricini Donovan) gut composition: implications for lignocellulose degradation.

Conversion of biomass such as lignocelluloses to an alternative energy source can contribute to sustainable development. Recently, biomass-degrading enzymes are reported to be common resources in insect-microbe interacting systems. Northeast India harbors ample sericigenous insect resources which are exploited for their silk products. Samia ricini Donovan is an economically important poly-phytophagous silkmoth capable of digesting foliage from different plant species, suggesting the versatility of a robust gut system. Here, a gut bacterial profile was determined by 16S rRNA gene characterization across the holometabolous life cycle during the summer and winter seasons, revealing 3 phyla, 13 families, and 22 genera. Comparative analysis among the seasonal gut isolates revealed a high diversity in summer, predominated by the genus Bacillus due to its high occurrence in all developmental stages. Shannon's diversity index demonstrated the second and fourth instars of summer as well as the fifth instar of winter to be relatively better developmental stages for gut bacteria assembly. Bacterial community shifts in concert to host developmental changes were found to be apparent between early instars and late instars in summer, which differed from those of winter. Forty-three and twenty-nine gut bacterial isolates were found to be cellulolytic and xylanolytic enzyme producers, respectively. The present results illustrate the gut microbiota of S. ricini over the seasons and support the holometabolous life cycle effect as the most likely factor shaping the gut bacterial microbiota. These findings may provide leads for the development of new cleaner and environmentally friendly lignocellulose-degrading enzymes.

Assessment of oily sludge biodegradation in lab scale composting and slurry bioreactor by bacterial consortium.

The present study aimed to investigate biodegradability of oily sludge in lab scale composting and slurry bioreactor using a potential bacterial consortium isolated from petroleum-contaminated sites. The consortium used in the study consisted of bacterial genera, including Enterobacter, Bacillus, Microbacterium, Alcaligenes Pseudomonas, Ochrobactrum, Micrococcus, and Shinella which were obtained after rigorous screening using different hydrocarbons. The meticulously designed lab scale composting experiments were carried out and showed that the combination of 10% oily sludge (A1) exhibited the highest total carbon (TC) removal, which was 40.33% within 90 days. To assess the composting experiments' efficiency, the first (k1) and second (k2) order rate constants were evaluated and was found to be 0.0004-0.0067 per day and second (k2) 0.0000008-0.00005 g/kg. day respectively. To further enhance the biodegradation rate of A1 combination, a slurry bioreactor was used. The maximum total petroleum hydrocarbon (TPH) removals in a slurry bioreactor for cycle-I and -II were 48.8% and 46.5%, respectively, on the 78th and 140th days of the treatment. The results obtained in the study will be a technological platform for the development of slurry phase treatment of petroleum waste in a sustainable and eco-friendly manner.

Occurrence, identification and characterization of diazotrophic bacteria from aerial roots of Rhynchostylis retusa (L.) Blume for plant growth-promoting activity.

In this study, the diversity of diazotrophic bacteria of orchid Rhynchostylis retusa (L.) Blume and its potential application in plant growth promotion were evaluated. About 183 nitrogen-fixing bacteria were isolated to screen various plant growth-promoting traits viz. phosphate solubilization,IAA, siderophore, HCN, biofilm and ammonia production. Based on 16S rRNA gene sequencing analysis Achromobacter, Arthrobacter, Acinetobacter, Bacillus, Brevibacterium, Curtobacterium, Erwinia, Kosakonia, Lysinibacillus, Klebseilla, Microbacterium, Mixta, Pantoea, Pseudomonas and Stenotrophomonas isolates were selected and showed positive results for PGP traits. Overall, genus Pantoea, Brevibacterium, Achromobacter, Arthrobacter, Klebsiella, Mixta, Bacillus, and Pseudomonas had the most pronounced PGP characteristics and acetylene reduction among the screened isolates. BOX PCR fingerprinting analysis showed variation in polymorphic banding patterns among diazotrophic strains. PCR amplification of nifH gene and the presence of 37 kDa nitrogenase reductase enzyme band in western blot indicated presence of nitrogenase activity. Our study showed that orchid R. retusa diazotroph interaction helps orchid plant to fix nitrogen, essential nutrients, and control pathogen entry. To the best of our knowledge, this is the first report on characterization of diazotrophic bacterial community from aerial roots of R. retusa.

De novogenomic analysis ofEnterobacter asburiaeEBRJ12, a plant growth-promoting rhizobacteria isolated from the rhizosphere of Phaseolus vulgarisL.

AIM: Environmental stresses such as water deficit induced stress are one of the major limiting factors in crop production. However, some plant growth-promoting rhizobacteria (PGPR) can promote plant growth in such adverse condition. Therefore, the objective was to isolate rhizospheric bacteria from Phaseolus vulgaris L. growing in a drought-affected soil and to analyze its plant growth promoting (PGP) efficacy to black gram (Vigna mungo L.) and Bhut jolokia (Capsicum chinense Jacq.). Whole-genome sequencing of the potential bacteria was targeted to analyze the genetic potential of the isolate as a plant growth-promoting agent. METHODS AND RESULTS: The isolate Enterobacter asburiae EBRJ12 was selected based on its PGP efficacy, which significantly improved plant growth and development. The genomic analysis revealed the presence of one circular chromosome of size 4.8 Mb containing 16 genes for osmotic stress regulation including osmotically inducible protein osmY, outer membrane protein A precursor ompA, aquaporin Z, and an operon for osmoprotectant ABC transporter yehZYXW. Moreover, the genome has a complete genetic cluster for biosynthesis of siderophore Enterobactin and siderophore Aerobactin.The PGP effects were verified with black gram and Bhut jolokia in pot experiments. The isolate significantly increased the shoot length by 35.0% and root length by 58.0% of black gram, while 41.0% and 57.0% of elevation in shoot and root length were observed in Bhut jolokia compared to non-inoculated plants. CONCLUSIONS: The EBRJ12 has PGP features that could improve the growth in host plants, and the genomic characterization revealed the presence of genetic potential for plant growth promotion.

An environment-benign approach of bamboo pulp bleaching using extracellular xylanase of strain Bacillus stratosphericus EB-11 isolated from elephant dung.

The use of microbial enzymes is highly encouraged in paper and pulp industries to reduce the excessive use of hazardous chemicals. During the study, xylanase of Bacillus stratosphericus EB-11 was characterized for pulp bleaching applications. The extracellular xylanase was produced under submerged fermentation using bamboo waste as a natural carbon source. There was fast cell division and enzyme production under optimized fermentation conditions in the bioreactor. The highest activity was 91,200U after 30 h of growth with Km and Vmax of 3.52 mg/mL and 391.5 µmol/min per mg respectively. The purified enzyme with molecular mass ~ 60 kDa had conferred positive activity on native PAGE. The strong inhibition by ethylenediaminetetraacetate and SDS showed the metallo-xylanase nature of the purified enzyme. The bacterial xylanase reduces the use of hydrogen peroxide by 0.4%. Similarly, biological oxygen demand and chemical oxygen demand were reduced by 42.6 and 35.2%. The xylanase-hydrogen peroxide combined treatment and conventional chlorine dioxide-alkaline (CDE1D1D2) bleaching showed almost similar improvement in physicochemical properties of bamboo pulp. Xylanase-peroxide bleaching reduces the lignin content to 4.95% from 13.32% unbleached pulp. This content after CDE1D1D2 treatment was 4.21%. The kappa number decreased from 15.2 to 9.46 with increasing the burst factor (15.51), crystallinity index (60.25%), viscosity (20.1 cp), and brightness (65.4%). The overall finding will encourage the development of new cleaner methods of bleaching in the paper and pulp industry.

Auxin biosynthesis by Microbacterium testaceum Y411 associated with orchid aerial roots and their efficacy in micropropagation.

Root-associated bacteria strongly affect plant growth and development by synthesizing growth regulators and stress-relieving metabolites. The present study is mainly focused on assessing aerial root-associated bacteria of Rhynchostylis retusa (L.) Blume is an endemic epiphytic orchid responsible for auxin production and influencing plant growth. A bacterial isolate, Microbacterium testaceum Y411, was found to be the most active producer of indole-3-acetic acid (IAA). The maximum IAA production (170µg/mL) was recorded with the bacterium at optimum process parameters such as pH 7, temperature 30°C, and tryptophan 1000 µg/mL in a culture medium for 48 h. The extracted auxin was purified and analyzed by FT-IR, HPLC, and HR-MS, indicating bacterial auxin has a similar mass value to 4-chloroindole-3-acetic acid auxin. Furthermore, the bacterial auxin was tested on in vitro propagation of orchid, Cymbidium aloifolium, and 90% seed germination was recorded in Murashige and Skoog's medium supplemented with bacterial auxin. The novel results obtained in this study are used for agricultural applications and the Microbacterium testaceum Y411 is a valuable biotechnological resource for a natural auxin.

Purification and characterization of cellulase produced by Novosphingobium sp. Cm1 and its waste hydrolysis efficiency and bio-stoning potential.

AIM: The aim of the study was to purify and characterize cellulase from a previously isolated Novosphingobium sp. strain Cm1 and to evaluate its waste hydrolysis and bio-stoning efficiency. MATERIALS AND METHODS: There is a growing demand for cellulase, a multipurpose enzyme widely used in industrial applications. Here, we purified cellulase from Novosphingobium sp. Cm1 by cellulose chromatography. SDS-PAGE revealed a molecular mass of 25 kDa. After 18-fold purification, the cellulase had an activity of 31.4 U/mg at pH of 5 and 40°C, and it retained activity at a wide range of pH and temperatures. The presence of Fe2+ and Co2+ boosted the enzyme activity by 57% and 25% respectively. The hydrolysing capacity of the strain towards cellulosic material was assessed for two paper types and the highest activity (2.6 ± 0.05 U/ml) was found with filter paper as the sole carbon source. Alterations in the structure of the papers as a result of bacterial hydrolysis were confirmed by scanning electron microscope and Fourier-transform infrared spectroscopy. The strain was also tested for its potential in various industrial applications and exhibited pectinolytic activity (6.78 ± 0.68 U/ml), xylanolytic activity (0.22 ± 0.14 U/ml) and bio-stoning ability. CONCLUSION: The highly active purified cellulase has a broad pH and temperature range. The strain possesses waste-hydrolysing ability, pectinolytic and xylanolytic ability along with bio-stoning capacity. SIGNIFICANCE AND IMPACT OF THE STUDY: The efficacy and versatility of the enzyme from Novosphingobium sp. Cm1 make it an excellent candidate for diverse industrial applications.

Production of cellulase by Novosphingobium sp. Cm1 and its potential application in lignocellulosic waste hydrolysis.

Management of lignocellulosic wastes in and around the municipality area requires special consideration. Continuous deposition of these wastes to the nearby areas led to gradual deterioration of the environment. The objective of this study was to produce cellulase from the bacteria isolated from the unexplored rainforest of NE-India for lignocellulosic waste hydrolysis. Based on carboxymethyl cellulose utilization and the congo red test, Novosphingobium sp. Cm1 was found to be the most promising strain out of 114 bacterial isolates and the strain was selected for further study. The optimization of the fermentative conditions for maximum enzyme activity was carried out using one factor-at-a-time strategy and the optimum pH, temperature and incubation time was recorded as pH 5, 37 °C and 96 h respectively. The maximum ß-1,4-endoglucanase activity was observed with 1.5% CMC (5.1 ± 0.05 U/mL) and 0.25% yeast extract (7.6 ± 0.72 U/mL). The bacterial waste hydrolysis ability was investigated using three wastes where vegetable waste showed maximum activity of 3.4 ± 0.48 U/mL. Bacterial interaction and waste utilization were verified using Scanning Electron Microscope and Fourier-Transform infrared spectroscopy analysis. The present study confirmed the promising ability of Novosphingobium sp. to waste hydrolysis. Further investigations may lead to new possibilities for low-cost enzyme production that will help to meet the rising cellulase demand.

Phylogenetic affiliation and antimicrobial effects of endophytic actinobacteria associated with medicinal plants: prevalence of polyketide synthase type II in antimicrobial strains.

The most diverse and versatile endophytic actinobacteria are relatively unexplored potential sources of bioactive metabolites useful for different medical, agricultural, and other commercial applications. Their diversity in symbiotic association with traditionally utilized medicinal plants of northeast India is scantly available. The present investigation assessed the genetic diversity of endophytic actinobacteria (n = 120) distributed around the root, stem, and leaf tissues of six selected medicinal plants (Emblica officinalis, Terminalia chebula, T. arjuna, Murraya koenigii, Rauwolfia serpentina, and Azadirachta indica) from three different protected areas of evergreen forest-the Gibbon Wildlife Sanctuary (GWS), the Kaziranga National Park (KNP), and the North East Ecological Park (NEEP) of Assam, India. The samples were collected in two seasons (summer and winter). The overall phylogenetic analysis showed significant genetic diversity with 18 distinct genera belonging to 12 families. Overall, the occurrence of Streptomyces genus was predominant across all three sampling sites (76.66%), in both the sampling season (summer and winter). Shannon's and Simpson's diversity estimates showed their presence at A. indica (1.496, 0.778), R. serpentina (1.470, 0.858), and E. officinalis (0.975, 0.353). Among the site sampled, GWS had the most diverse community of actinobacteria (Shannon = 0.86 and Simpson = 0.557). The isolates were antagonistically more active against the investigated plant pathogenic bacteria than fungal pathogens. Further analysis revealed the prevalence of polyketide synthase genes (PKS) type II (84%) and PKS type I (16%) in the genome of the antimicrobial isolates. The overall findings confirmed the presence of biosynthetically active diverse actinobacterial members in the selected medicinal plants which offer potential opportunities towards the exploration of biologically active compounds.

Mapping the Bacterial Community in Digboi Oil Refinery, India by High-Throughput Sequencing Approach.

Microbial enhanced oil recovery (MOER) is a technique which uses microbes to enhance the oil recovery process. This technique is advantageous to enhance oil recovery (EOR). In this study, we analyzed the bacterial communities of Digboi oil refinery and its surroundings using Illumina MiSeq sequencing platform. A total of 12 samples were analyzed, 6 from inside the refinery areas and another 6 from the township areas. Alpha diversity studies indicated that diversity of bacterial communities in township area was higher than the refinery areas except for Sample 1. Sample 9 from the nearby pond of Digboi Centenary Park was more diverse in community composition. Proteobacteria was found to be most dominant phylum. Mantel test indicated that environmental factors had negative influence over the bacterial community structure. Among the environmental factors Fe was least significant (r2 = 0.368) as indicated by canonical correspondence analysis.

Author Correction: Alleviation of drought stress in pulse crops with ACC deaminase producing rhizobacteria isolated from acidic soil of Northeast India.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Author Correction: Insights into the functionality of endophytic actinobacteria with a focus on their biosynthetic potential and secondary metabolites production.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Alleviation of drought stress in pulse crops with ACC deaminase producing rhizobacteria isolated from acidic soil of Northeast India.

The agricultural crops are often affected by the scarcity of fresh water. Seasonal drought is a major constraint on Northeast Indian agriculture. Almost 80% of the agricultural land in this region is acidic and facing severe drought during the winter period. Apart from classical breeding and transgenic approaches, the application of plant-growth-promoting bacteria (PGPB) is an alternative strategy for improving plant fitness under stressful conditions. The 1-aminocyclopropane-1-carboxylate (ACC) deaminase-producing PGPB offer drought stress tolerance by regulating plant ethylene levels. The aim of the present study was to evaluate the consortium effect of three ACC-deaminase producing rhizobacteria - Ochrobactrum pseudogrignonenseRJ12, Pseudomonas sp.RJ15 and Bacillus subtilisRJ46 on drought stress alleviation in Vigna mungo L. and Pisum sativum L. Consortium treatment significantly increase seed germination percentage, root length, shoot length, and dry weight of treated plants. An elevated production of reactive oxygen species scavenging enzymes and cellular osmolytes; higher leaf chlorophyll content; increase in relative water content and root recovery intension were observed after consortium treatment in comparison with the uninoculated plants under drought conditions. The consortium treatment decreased the ACC accumulation and down-regulated ACC-oxidase gene expression. This consortium could be an effective bio-formulator for crop health improvement in drought-affected acidic agricultural fields.

Insights into the functionality of endophytic actinobacteria with a focus on their biosynthetic potential and secondary metabolites production.

Endophytic actinobacteria play an important role in growth promotion and development of host plant by producing enormous quantities of novel bioactive natural products. In the present investigation, 169 endophytic actinobacteria were isolated from endospheric tissues of Rhynchotoechum ellipticum. Based on their antimicrobial potential, 81 strains were identified by 16rRNA gene analysis, which were taxonomically grouped into 15 genera. All identified strains were screened for their plant growth promoting attributes and, for the presence of modular polyketide synthases (PKSI, PKSII and nonribosomal peptide synthetase (NRPS) gene clusters to correlate the biosynthetic genes with their functional properties. Expression studies and antioxidant potential for four representative strains were evaluated using qRT-PCR and DPPH assay respectively. Additionally, six antibiotics (erythromycin, ketoconazole, fluconazole, chloramphenicol, rifampicin and miconazole) and nine phenolic compounds (catechin, kaempferol, chebulagic acid, chlorogenic acid, Asiatic acid, ferulic acid, arjunic acid, gallic acid and boswellic acid) were detected and quantified using UHPLC-QqQLIT-MS/MS. Furthermore, three strains (BPSAC77, 121 and 101) showed the presence of the anticancerous compound paclitaxel which was reported for the first time from endophytic actinobacteria. This study provides a holistic picture, that endophytic actinobacteria are rich bacterial resource for bioactive natural products, which has a great prospective in agriculture and pharmaceutical industries.

Rhizospheric Bacterial Community of Endemic Rhododendron arboreum Sm. Ssp. delavayi along Eastern Himalayan Slope in Tawang.

Information on rhizosphere microbiome of endemic plants from high mountain ecosystems against those of cultivated plantations is inadequate. Comparative bacterial profiles of endemic medicinal plant Rhododendron arboreum Sm. subsp. delavayi rhizosphere pertaining to four altitudinal zonation Pankang Thang (PTSO), Nagula, Y-junction and Bum La (Indo-China border; in triplicates each) along cold adapted Eastern slope of Himalayan Tawang region, India is described here. Significant differences in DGGE profile between below ground bulk vs. rhizospheric community profile associated with the plant was identified. Tagged 16S amplicon sequencing from PTSO (3912 m) to Bum La (4509 m), revealed that soil pH, total nitrogen (TN), organic matter (OM) significantly influenced the underlying bacterial community structure at different altitudes. The relative abundance of Acidobacteria was inversely related to pH, as opposed to TN which was positively correlated to Acidobacteria and Proteobacteria abundance. TN was also the significant predictor for less abundant taxonomic groups Chloroflexi, Gemmatimonadetes, and Nitrospirae. Bum La soil harbored less bacterial diversity compared to other sites at lower altitudes. The most abundant phyla at 3% genetic difference were Acidobacteria, Actinobacteria, and Proteobacteria amongst others. Analysis of similarity indicated greater similarity within lower altitudinal than higher altitudinal group (ANOSIM, R = 0.287, p = 0.02). Constraining the ordination with the edaphic factor explained 83.13% of variation. Unique phylotypes of Bradyrhizobium and uncultured Rhizobiales were found in significant proportions at the four regions. With over 1% relative abundance Actinobacteria (42.6%), Acidobacteria (24.02%), Proteobacteria (16.00%), AD3 (9.23%), WPS-2 (5.1%), and Chloroflexi (1.48%) dominated the core microbiome.

A Novel Triculture System (CC3) for Simultaneous Enzyme Production and Hydrolysis of Common Grasses through Submerged Fermentation.

The perennial grasses are considered as a rich source of lignocellulosic biomass, making it a second generation alternative energy source and can diminish the use of fossil fuels. In this work, four perennial grasses Saccharum arundinaceum, Panicum antidotale, Thysanolaena latifolia, and Neyraudia reynaudiana were selected to verify their potential as a substrate to produce hydrolytic enzymes and to evaluate them as second generation energy biomass. Here, cellulase and hemi-cellulase producing three endophytic bacteria (Burkholderia cepacia BPS-GB3, Alcaligenes faecalis BPS-GB5 and Enterobacter hormaechei BPS-GB8) recovered from N. reynaudiana and S. arundinaceum were selected to develop a triculture (CC3) consortium. During 12 days of submerged cultivation, a 55-70% loss in dry weight was observed and the maximum activity of ß-glucosidase (5.36-12.34 IU) and Xylanase (4.33 to 10.91 IU) were observed on 2nd and 6th day respectively, whereas FPase (0.26 to 0.53 IU) and CMCase (2.31 to 4.65 IU) showed maximum activity on 4th day. Around 15-30% more enzyme activity was produced in CC3 as compared to monoculture (CC1) and coculture (CC2) treatments, suggested synergetic interaction among the selected three bacterial strains. Further, the biomass was assessed using Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). The FTIR analysis provides important insights into the reduction of cellulose and hemicellulose moieties in CC3 treated biomass and SEM studies shed light into the disruption of surface structure leading to access of cellulose or hemicelluloses microtubules. The hydrolytic potential of the CC3 system was further enhanced due to reduction in lignin as evidenced by 1-4% lignin reduction in biomass compositional analysis. Additionally, laccase gene was detected from A. faecalis and E. hormaechei which further shows the laccase production potential of the isolates. To our knowledge, first time we develop an effective endophytic endogenous bacterial triculture system having potential for the production of extracellular enzymes utilizing S. arundinaceum and N. reynaudiana as lignocellulosic feedstock.

Distribution and Identification of Endophytic Streptomyces Species from Schima wallichii as Potential Biocontrol Agents against Fungal Plant Pathogens.

The prospective of endophytic microorganisms allied with medicinal plants is disproportionally large compared to those in other biomes. The use of antagonistic microorganisms to control devastating fungal pathogens is an attractive and eco-friendly substitute for chemical pesticides. Many species of actinomycetes, especially the genus Streptomyces, are well known as biocontrol agents. We investigated the culturable community composition and biological control ability of endophytic Streptomyces sp. associated with an ethanobotanical plant Schima wallichi. A total of 22 actinobacterial strains were isolated from different organs of selected medicinal plants and screened for their biocontrol ability against seven fungal phytopathogens. Seven isolates showed significant inhibition activity against most of the selected pathogens. Their identification based on 16S rRNA gene sequence analysis, strongly indicated that all strains belonged to the genus Streptomyces. An endophytic strain BPSAC70 isolated from root tissues showed highest percentage of inhibition (98.3 %) against Fusarium culmorum with significant activity against other tested fungal pathogens. Phylogenetic analysis based on 16S rRNA gene sequences revealed that all seven strains shared 100 % similarity with the genus Streptomyces. In addition, the isolates were subjected to the amplification of antimicrobial genes encoding polyketide synthase type I (PKS-I) and nonribosomal peptide synthetase (NRPS) and found to be present in most of the potent strains. Our results identified some potential endophytic Streptomyces species having antagonistic activity against multiple fungal phytopathogens that could be used as an effective biocontrol agent against pathogenic fungi.