Bacillus Altitudinis Research Articles

Modulation in Plant Growth and Drought Tolerance of Wheat Crop upon Inoculation of Drought-tolerant-Bacillus Species Isolated from Hot Arid Soil of India

Drought is one of the most detrimental environmental stressors to plants with the potential to decrease crop yields and affect agricultural sustainability. Native bacteria with beneficial traits enhance plant growth and help avoid and reverse the effects of drought in plants to a greater extent. In the present study, we aimed to ( i ) isolate drought-tolerant Bacillus isolates from the rhizosphere soil of wheat crop grown at different locations in Jaisalmer district, Rajasthan state and (ii) further evaluate their ability to enhance plant growth and induce drought tolerance in wheat ( Var. HD-2967) grown under drought stress conditions. Of more than 100 isolates, two putative Bacillus isolates capable of tolerating 30 % polyethylene glycol-6000 (PEG-6000) [equivalent to -9.80 MPa (Megapascal)] were identified as Bacillus altitudinis DT-89 and Bacillus paramycoides DT-113. These isolates exhibited different plant growth promoting (PGP) attributes such as phosphate solubilization, and production of siderophore, exopolysaccharide, ammonia, indole acetic acid and cytokinin at low osmotic stress of 10% PEG-6000 but shown variable response at higher osmotic stress particularly at 30% PEG-6000. However, they did not show any antifungal activity and one isolate was negative for phosphate solubilization. Of two strains, B. altitudinis DT-89 function more prominently with respect to plant growth promotion and drought tolerance to plant in the early stage but protective traits of B. paramycoides DT-113 was more prominent after 75 days as evident by increased EPS (164%), root dry weight (144.44%), chlorophyll content (90.26%), SOD (389%) and proline (99.3%). The results support both the strains as a potential candidate to alleviate drought stress and enhance plant growth in the drought regions.

Waterborne Antifouling Paints Containing Nanometric Copper and Silver against Marine Bacillus Species.

Due to the concern to find an alternative to reduce the colonization (microfouling and macrofouling) or the biocorrosion of surfaces submerged for long periods in water, we evaluated the antifouling activity of a commercial paint added with silver nanoparticles (AgNP's) and copper nanoparticles (CuNP's), beside copper-soybean chelate, by electrolytic synthesis, using them in low concentrations (6.94E − 04 mg Ag g−1 paint, 9.07E − 03 mg Cu g−1 paint, and 1.14E − 02 mg Cu g−1 paint, respectively). The test for paint samples was carried out by JIS Z2801-ISO 22196 for periods of initial time, 6 months, and 12 months, against three bacterial strains of marine origin, Bacillus subtilis, Bacillus pumilus, and Bacillus altitudinis. It was possible to demonstrate, according to the standard, that the sample with the greatest antimicrobial activity was the copper-soybean chelate against two of the three strains studied (B. pumilus with R = 2.11 and B. subtilis with R = 2.41), which represents more than 99% of bacterial inhibition. Therefore, we considered a novel option for inhibiting bacterial growth with nanoparticles as antifouling additives.

An Insight into Algicidal Characteristics of Bacillus Altitudinis G3 from Dysfunctional Photosystem and Overproduction of Reactive Oxygen Species

An Insight into Algicidal Characteristics of Bacillus Altitudinis G3 from Dysfunctional Photosystem and Overproduction of Reactive Oxygen Species

Degradation of lignin by Bacillus altitudinis SL7 isolated from pulp and paper mill effluent.

Lignin is a major by-product of pulp and paper industries, and is resistant to depolymerization due to its heterogeneous structure. Degradation of lignin can be achieved by the use of potential lignin-degrading bacteria. The current study was designed to evaluate the degradation efficiency of newly isolated Bacillus altitudinis SL7 from pulp and paper mill effluent. The degradation efficiency of B. altitudinis SL7 was determined by color reduction, lignin content, and ligninolytic activity from degradation medium supplemented with alkali lignin (3 g/L). B. altitudinis SL7 reduced color and lignin content by 26 and 44%, respectively, on the 5th day of incubation, as evident from the maximum laccase activity. Optimum degradation was observed at 40 °C and pH 8.0. FT-IR spectroscopy and GC-MS analysis confirmed lignin degradation by emergence of the new peaks and identification of low-molecular-weight compounds in treated samples. The identified compounds such as vanillin, 2-methyoxyhenol, 3-methyl phenol, oxalic acid and ferulic acid suggested the degradation of coniferyl and sinapyl groups of lignin. Degradation efficiency of B. altitudinis SL7 towards high lignin concentration under alkaline pH indicated the potential application of this isolate in biological treatment of the lignin-containing effluents.

Intestinal microbiota modulation and improved growth in pigs with post-weaning antibiotic and ZnO supplementation but only subtle microbiota effects with Bacillus altitudinis

The objective was to evaluate the effect of dietary Bacillus altitudinis spore supplementation during day (D)0–28 post-weaning (PW) and/or D29–56 PW compared with antibiotic and zinc oxide (AB + ZnO) supplementation on pig growth and gut microbiota. Eighty piglets were selected at weaning and randomly assigned to one of five dietary treatments: (1) negative control (Con/Con); (2) probiotic spores from D29–56 PW (Con/Pro); (3) probiotic spores from D0–28 PW (Pro/Con); (4) probiotic spores from D0–56 PW (Pro/Pro) and (5) AB + ZnO from D0–28 PW. Overall, compared with the AB + ZnO group, the Pro/Con group had lower body weight, average daily gain and feed intake and the Pro/Pro group tended to have lower daily gain and feed intake. However, none of these parameters differed between any of the probiotic-treated groups and the Con/Con group. Overall, AB + ZnO-supplemented pigs had higher Bacteroidaceae and Prevotellaceae and lower Lactobacillaceae and Spirochaetaceae abundance compared to the Con/Con group, which may help to explain improvements in growth between D15–28 PW. The butyrate-producing genera Agathobacter, Faecalibacterium and Roseburia were more abundant in the Pro/Con group compared with the Con/Con group on D35 PW. Thus, whilst supplementation with B. altitudinis did not enhance pig growth performance, it did have a subtle, albeit potentially beneficial, impact on the intestinal microbiota.

Effects of host‐associated probioticBacillus altitudinisB61‐34b on growth performance, immune response and disease resistance of Nile tilapia (Oreochromis niloticus) raised under biofloc system

This investigation was performed to assess the influences of the host-associated probiotic Bacillus altitudinis B61-34b (BAA) on growth, immune response and disease resistance to Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus). Fish (25.50 ± 0.52 g fish−1) were distributed into 15 aquaria (150 litres tank−1) at density of 20 fish tank−1. The fish were fed basal diets with BAA inclusions in different concentrations: 0 (BAA1—Control), 106 (BAA2), 107 (BAA3), 108 (BAA4) and 109 (BAA5) CFU ml−1 for 8 weeks. A completely randomized design with three replications was used. The results indicated that BAA administration generated greater (p < 0.05) skin mucus and peroxidase activities compared to the control, with maximum levels recorded in BAA4 and BAA5. Higher serum immunities, such as serum lysozyme, serum peroxidase, complement, phagocytosis and respiratory burst activities were observed in the BAA4 and BAA5 dietary groups compared to the control, BAA1, BAA2 and BAA3 diets. The relative percentage of survival, growth performance and FCR were also significantly higher (p < 0.05) in the BAA4 and BAA5 groups. In conclusion, dietary supplementation of B. altitudinis at 108 CFU ml−1 can improve Nile tilapia's growth performance, skin mucus and serum immunities, and disease resistance.

Inter-Genera Colonization of Ocimum tenuiflorum Endophytes in Tomato and Their Complementary Effects on Na+/K+ Balance, Oxidative Stress Regulation, and Root Architecture Under Elevated Soil Salinity.

Endophytic bacilli of ethano-botanical plant Ocimum tenuiflorum were screened for salt stress-alleviating traits in tomato. Four promising O. tenuiflorum endophytes (Bacillus safensis BTL5, Bacillus haynesii GTR8, Bacillus paralicheniformis GTR11, and Bacillus altitudinis GTS16) were used in this study. Confocal scanning laser microscopic studies revealed the inter-genera colonization of O. tenuiflorum endophytes in tomato plants, giving insights for widening the applicability of potential endophytes to other crops. Furthermore, in a pot trial under 150 mM NaCl concentration, the inoculated endophytes contributed in reducing salt toxicity and improving recovery from salt-induced oxidative stress by different mechanisms. Reduction in reactive oxygen species (ROS) (sub-cellular H2O2 and superoxide) accumulation was observed besides lowering programmed cell death and increasing chlorophyll content. Endophyte inoculation supplemented the plant antioxidant enzyme system via the modulation of enzymatic antioxidants, viz., peroxidase, ascorbate peroxidase, superoxide dismutase, and catalase, apart from increasing proline and total phenolics. Antioxidants like proline have dual roles of antioxidants and osmoregulation, which might also have contributed to improved water relation under elevated salinity. Root architecture, viz., root length, projection area, surface area, average diameter, tips, forks, crossings, and the number of links, was improved upon inoculation, indicating healthy root growth and enhanced nutrient flow and water homeostasis. Regulation of Na+/K+ balance and water homeostasis in the plants were also evident from the modulation in the expression of abiotic stress-responsive genes, viz., LKT1, NHX1, SOS1, LePIP2, SlERF16, and SlWRKY39. Shoot tissues staining with light-excitable Na+ indicator Sodium GreenTM Tetra (tetramethylammonium) salt showed low sodium transport and accumulation in endophyte-inoculated plants. All four endophytes exhibited different mechanisms for stress alleviation and indicated complementary effects on plant growth. Furthermore, this could be harnessed in the form of a consortium for salt stress alleviation. The present study established inter-genera colonization of O. tenuiflorum endophytes in tomato and revealed its potential in maintaining Na+/K+ balance, reducing ROS, and improving root architecture under elevated salinity.

Diverse thermophilic Bacillus species with multiple biotechnological activities are associated within the Egyptian soil and compost samples.

Thermophilic strains of Bacillus can express enzymes of higher thermal stability, which allows carrying out industrial fermentations under higher temperatures. This lowers the contamination potential, accelerates mixing rates and facilitates the recovery of fermentation end products. The present study was thus designed to isolate and characterize thermophilic Bacillus cultures from soil and compost samples. Forty-two thermophilic Bacillus isolates could be identified employing morphological, physiological and the 16S rRNA gene sequencing analyses. The isolates showed a high degree of biological diversity involving 13 Bacillus species and 1 subspecies but were dominated by Bacillus licheniformis. Phylogenetic analysis of B. licheniformis isolates based on the DNA sequencing of gyrA and rpoB genes presented them in two main genetic groups. Isolates of five thermophilic species including B. licheniformis, Bacillus altitudinis, Bacillus paralicheniformis, Bacillus subtilis and Bacillus thermoamylovorans showed multiple activities to degrade all of cellulose, hemicellulose and lignin. Those multifunctional thermophilic Bacillus isolates can be harnessed in the degradation of plant wastes for the production of biofuels and compost.

Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum.

Bioconversion of lignocellulose into renewable energy and commodity products faces a major obstacle of inefficient saccharification due to its recalcitrant structure. In nature, lignocellulose is efficiently degraded by some insects, including termites and beetles, potentially due to the contribution from symbiotic gut bacteria. To this end, the presented investigation reports the isolation and characterization of cellulolytic bacteria from the gut system of red flour beetle, Tribolium castaneum. Out of the 15 isolated bacteria, strain RSP75 showed the highest cellulolytic activities by forming a clearance zone of 28 mm in diameter with a hydrolytic capacity of ~4.7. The MALDI-TOF biotyping and 16S rRNA gene sequencing revealed that the strain RSP75 belongs to Bacillus altitudinis. Among the tested enzymes, B. altitudinis RSP75 showed maximum activity of 63.2 IU/mL extract for xylanase followed by β-glucosidase (47.1 ± 3 IU/mL extract) which were manifold higher than previously reported activities. The highest substrate degradation was achieved with wheat husk and corn cob powder which accounted for 69.2% and 54.5%, respectively. The scanning electron microscopy showed adhesion of the bacterial cells with the substrate which was further substantiated by FTIR analysis that depicted the absence of the characteristic cellulose bands at wave numbers 1247, 1375, and 1735 cm−1 due to hydrolysis by the bacterium. Furthermore, B. altitudinis RSP75 showed co-culturing competence with Saccharomyces cerevisiae for bioethanol production from lignocellulose as revealed by GC-MS analysis. The overall observations signify the gut of T. castaneum as a unique and impressive reservoir to prospect for lignocellulose-degrading bacteria that can have many biotechnological applications, including biofuels and biorefinery.

Biodegradation of organophosphorus pesticide profenofos by the bacterium Bacillus sp. PF1 and elucidation of initial degradation pathway

ABSTRACT Among the organophosphate pesticides, the wide and indiscriminate use of profenofos (PFF) in agricultural and horticultural crops has resulted in serious environmental and animal health concerns and therefore demands an urgent need to develop a biological solution for its effective removal from the environment. For the bioremediation of PFF, a strain PF1, capable of utilizing profenofos as the sole source of carbon and energy was isolated from the soil samples of apple orchards of Shimla region of Himachal Pradesh, India. Based on the biochemical, FAME, and 16S rRNA gene analysis the bacterium PF1 was identified as Bacillus altitudinis (GenBank: MH986176). The strain was able to degrade 50μg mL−1 PFF up to 93% within 30 days of incubation at 28°C, pH 7.0. A linear regression analysis performed on the data-set revealed the statistical significance of the relationship between the growth of the bacterial population and the degradation of pesticides. The compound 4-Bromo-2-chlorophenol (BCP) was detected as one of the pathway metabolites which further were completely degraded to lower pathway metabolites. A probable PFF degradation pathway has been proposed which follows the path from PFF to BCP and ultimately enters into the TCA cycle. To the best of our knowledge, this is the first report of PFF biodegradation by any Bacillus species of western Himalayan origin exhibiting close phylogenetic association with Bacillus altitudinis. This indigenous bacterium can be useful to bio-remediate the PFF contaminated soil as this pesticide is extensively used in the different horticulture fields in Himachal Pradesh, India.

Characterisation of Plant Growth-Promoting Endophytic Bacteria from Sugarcane and Their Antagonistic Activity against Fusarium moniliforme.

The use of endophytic bacteria in agriculture provides an effective way of improving crop yield and significantly reducing chemical usage, such as fungicides. This research was conducted to explore endophytic bacteria with plant growth promotion (PGP) and antifungal activities against Fusarium moniliforme AIT01. In this study, we obtained 52 isolates of endophytic bacteria associated with the roots and stems of sugarcane from Nakhon Ratchasima province, Thailand. In vitro antagonistic activity test showed that 14 out of 52 isolates had antagonistic activity against the fungal pathogen F. moniliforme AIT01. These antagonistic endophytic bacteria were identified as belonging to six different species as follows: Nguyenibacter vanlangensis, Acidomonas methanolica, Asaia bogorensis, Tanticharoenia aidae, Burkholderia gladioli and Bacillus altitudinis based on phenotypic characteristics, along with phylogenetic analysis of their 16S rRNA gene sequences. Seven isolates effectively inhibited F. moniliforme AIT01 mycelial growth by up to 40%. The volatile compounds of six isolates reduced the growth of F. moniliforme AIT01 by over 23%. Moreover, riceberry rice seedlings previously treated with B. gladioli CP28 were found to strongly reduce infection with phytopathogen by 80% in comparison to the non-treated control. Furthermore, the isolates also showed relevant PGP features, including ammonia production, zinc and phosphate solubilisation, auxin and siderophore biosynthesis. These results demonstrated that the tested endophytic bacteria could be successfully utilised as a source of PGP and biocontrol agent to manage diseases caused by F. moniliforme.

Microbial-Assisted Wheat Iron Biofortification Using Endophytic Bacillus altitudinis WR10.

Microbial-assisted biofortification attracted much attention recently due to its sustainable and eco-friendly nature for improving nutrient content in wheat. An endophytic strain Bacillus altitudinis WR10, which showed sophistical regulation of iron (Fe) homeostasis in wheat seedlings, inspired us to test its potential for enhancing Fe biofortification in wheat grain. In this study, assays in vitro indicated that WR10 has versatile plant growth-promoting (PGP) traits and bioinformatic analysis predicted its non-pathogenicity. Two inoculation methods, namely, seed soaking and soil spraying, with 107 cfu/ml WR10 cells were applied once before sowing of wheat (Triticum aestivum L. cv. Zhoumai 36) in the field. After wheat maturation, evaluation of yield and nutrients showed a significant increase in the mean number of kernels per spike (KPS) and the content of total nitrogen (N), potassium (K), and Fe in grains. At the grain filling stage, the abundance of Bacillus spp. and the content of N, K, and Fe in the root, the stem, and the leaf were also increased in nearly all tissues, except Fe in the stem and the leaf. Further correlation analysis revealed a positive relationship between the total abundance of Bacillus spp. and the content of N, K, and Fe in grains. Seed staining confirmed the enhanced accumulation of Fe, especially in the embryo and the endosperm. Finally, using a hydroponic coculture model, qPCR quantification indicated effective colonization, internalization, translocation, and replication of strain WR10 in wheat within 48 h. Collectively, strain WR10 assisted successful Fe biofortification in wheat in the field, laying a foundation for further large-scale investigation of its applicability and effectiveness.

Phylogeny of the Bacillus altitudinis Complex and Characterization of a Newly Isolated Strain with Antilisterial Activity

Bacillus strain UTK D1-0055 was isolated from a laboratory environment and appeared to have antilisterial activity. The genome was sequenced, the strain was identified as Bacillus altitudinis, and a high-quality complete annotated genome was produced. The taxonomy was evaluated for this and related Bacillus species (B. aerophilus, B. pumilus, B. safensis, B. stratosphericus, and B. xiamenensis) because the taxonomy is unclear and contains errors in public databases such as NCBI. The included strains grouped into seven clusters based on average nucleotide identity. Strains designated as B. aerophilus, B. altitudinis, and B. stratosphericus grouped together in the cluster containing the B. altitudinis type strain, suggesting that these three species should be considered a single species, B. altitudinis. The antimicrobial activity of UTK D1-0055 was evaluated against a panel of 15 Listeria strains (nine Listeria monocytogenes serotypes, Listeria innocua, and Listeria marthii), other foodborne pathogens (six Salmonella enterica serotypes and Escherichia coli), and three representative fungi (Saccharomyces cerevisiae, Botrytis cinerea, and Hyperdermium pulvinatum). Antibacterial activity was observed against all Listeria strains, but no antibacterial effects were found against the other tested bacterial and fungal strains. Biosynthetic gene clusters were identified in silico that may be related to the observed antibacterial activity, and these clusters included genes that putatively encode bacteriocins and nonribosomally synthesized peptides. The B. altitudinis strain identified in this investigation had a broad range of antilisterial activity, suggesting that it and other related strains may be useful for biocontrol in the food industry.

Biocontrol of Pythium aphanidermatum causing soft rot in ginger with biosurfactant produced by a rhizospheric Bacillus species

Plant growth-promoting rhizobacteria benefit plants by producing phytohormones and acquiring vital nutrients from soil. They also form a shield against potential phytopathogens with the production of antimicrobial compounds such as bacteriocin, biosurfactants etc. In the present study, among various rhizobacteria isolated from ginger rhizosphere, the isolate identified as Bacillus altitudinis VS7 that showed a potent biosurfactant producing ability in the preliminary tests was selected for the further study. Compositional analyses indicated that the biosurfactant produced by VS7 was a surfactin like compound, a cyclic lipopeptide. In both microtitre plate and mycelium growth inhibition studies, the biosurfactant showed dose dependent antagonistic activities against Pythium aphanidermatum that causes soft rot in ginger. Although there are reports of biocontrol activity of various rhizobacteria against soft rot in ginger, this is the firstinformation regarding biocontrol activity of a biosurfactant produced by the rhizobacterium B. altitudinis VS7 against P. aphanidermatum.

Identification and Genomic Characterization of Pathogenic Bacillus altitudinis from Common Pear Trees in Morocco

Bacterial burn is one of the major diseases affecting pear trees worldwide, with serious impacts on producers and economy. In Morocco, several pear trees (Pyrus communis) have shown leaf burns since 2015. To characterize the causal agent of this disease, we isolated fourteen bacterial strains from different parts of symptomatic pear trees (leaves, shoots, fruits and flowers) that were tested in planta for their pathogenicity on Louise bonne and Williams cultivars. The results showed necrotic lesions with a significant severity range from 47.63 to 57.77% on leaves of the Louise bonne cultivar inoculated with isolate B10, while the other bacterial isolates did not induce any disease symptom. 16S rRNA gene sequencing did not allow robust taxonomic discrimination of the incriminated isolate. Thus, we conducted whole-genome sequencing (WGS) and phylogenetic analyzes based on gyrA, gyrB and cdaA gene sequences, indicating that this isolate belongs to the Bacillus altitudinis species. This taxonomic classification was further confirmed by the Average Nucleotide Identity (ANI) and the in silico DNA-DNA hybridization (isDDH) analyzes compared to sixty-five Bacillus spp. type strains. The genome was mined for genes encoding carbohydrate-active enzymes (CAZymes) known to play a role in the vegetal tissue degradation. 177 candidates with functions that may support the in planta phytopathogenicity results were identified. To the best of our knowledge, this is the first data reporting B. altitudinis as agent of leaf burn in P. communis in Morocco. Our dataset will improve our knowledge on spread and pathogenicity of B. altitudinis genotypes that appears as emergent phytopathogenic agent, unveiling virulence factors and their genomic location (i.e., within genomic islands or the accessory genome) to induce trees disease.

Evaluation of the Potential Role of Bacillus altitudinis MT422188 in Nickel Bioremediation from Contaminated Industrial Effluents

The incessant pervasiveness of heavy metals in the environment is one of the precursory factors of pollution. This research study was endeavored upon to investigate the bioremediation potential of a nickel (Ni)-resistant bacterial isolate, identified as Bacillus altitudinis MT422188, whose optimum growth parameters were demonstrated at pH 7, temperature 32 °C, and 1 mM phosphate. Minimal Inhibitory Concentration (MIC) and EC50 for Ni were observed to be 20 and 11.5 mM, respectively, whereas the cross heavy-metal resistance was discerned as Cu2+ (25 mM) &gt; Zn2+ (15 mM) &gt; Cr6+ (10 mM) &gt; Pb2+ (5 mM) &gt; Co2+ (8 mM) &gt; Cd2+ (3 mM) &gt; Hg2+ (0 mM). Ni biosorption studies by live and heat-killed bacterial cells were suggestive of Ni uptake being facilitated by an ATP-independent efflux system. A pilot-scale study displayed the effective removal of Ni (70 mg/L and 85 mg/L) at 4- and 8-day intervals, respectively. Moreover, chemotaxis and motility assays indicated the role of Ni as a chemoattractant for bacterial cells. The presence of Ni reduced the GR (0.001 ± 0.003 Ug−1FW), POX (0.001 ± 0.001 Ug−1FW), and SOD (0.091 ± 0.003 Ug−1FW) activity, whereas Sodium dodecyl sulphate—Polyacrylamide gel electrophoresis (SDS-PAGE) revealed the presence of metallothionein (60 kDa). Kinetic and isotherm studies suggested a pseudo second-order and Freundlich model to be better fitted for our study. The thermodynamic parameters (∆H° = 3.0436 kJ/mol, ∆S° = 0.0224 kJ/mol/K) suggested the process to be endothermic, spontaneous, and favorable in nature. FTIR analysis elucidated the interaction of hydroxyl and carboxyl groups with Ni. Scanning Electron Microscopy (SEM) and Energy Dispersive X-Ray Spectroscopy (EDS) demonstrated changes in the morphological and elemental composition of the bacterial cells, which affirmed their interaction with Ni during biosorption. In summary, our study concludes the efficient role of Bacillus altitudinis MT422188 in removing Ni from polluted industrial effluents.

Characterization of endospore-forming bacteria producing extracellular enzymes isolated from Djurdjura Mountains in Algeria

Biodiversity in mountains in Algeria appears scanty and has not been thoroughly investigated. However, the mountain soil has been shown as an almost entire reserve of novel enzymes with interesting properties for industrial and environmental applications. In the present study, 30 bacterial strains were isolated from the Djurdjura Mountains in Kabylia (Algeria) and were studied for their ability to produce enzymes to be possibly used in biotechnological processes such as amylase, protease, and chitinase. The characterization of these isolates was carried out using morphological, physiological, and biochemical characteristics. All the data obtained with regards to the phenotypical properties of the isolates, confirmed that the strains belonged to the Bacillus group. In addition, the 16S rRNA gene of the two retained strains KA15 and LK-DZ15 was also amplified and sequenced. Phylogenetic tree was, afterwards, constructed. The nucleotide sequences and blast analyses confirmed that the KA15 and LK-DZ15 strains were closely related to those of the Bacillus altitudinis (accession n°.: MK874318) and Paenibacillus timonensis (accession n°.: MK734103) strains. The presence of amylases, proteases, and chitinases in KA15 and LK-DZ15 isolates are an indicator of their pivotal application in a variety of biotechnological processes.

Endophytic Bacillus altitudinis Strain Uses Different Novelty Molecular Pathways to Enhance Plant Growth.

The identification and use of endophytic bacteria capable of triggering plant growth is an important aim in sustainable agriculture. In nature, plants live in alliance with multiple plant growth-promoting endophytic microorganisms. In the current study, we isolated and identified a new endophytic bacterium from a wild plant species Glyceria chinensis (Keng). The bacterium was designated as a Bacillus altitudinis strain using 16S rDNA sequencing. The endophytic B. altitudinis had a notable influence on plant growth. The results of our assays revealed that the endophytic B. altitudinis raised the growth of different plant species. Remarkably, we found transcriptional changes in plants treated with the bacterium. Genes such as maturase K, tetratricopeptide repeat-like superfamily protein, LOB domain-containing protein, and BTB/POZ/TAZ domain-containing protein were highly expressed. In addition, we identified for the first time an induction in the endophytic bacterium of the major facilitator superfamily transporter and DNA gyrase subunit B genes during interaction with the plant. These new findings show that endophytic B. altitudinis could be used as a favourable candidate source to enhance plant growth in sustainable agriculture.

Bacillus altitudinis MT422188: a potential agent for zinc bioremediation

Anthropogenic processes are one of the main contributors for the presence of heavy metals in air, land and water sources, leading to environmental pollution. In the present study, Bacillus altitudinis MT422188 was used for determining its potential role in zinc bioremediation. After the bacterium was isolated and characterized, its optimum growth conditions were determined. The growth parameters, antioxidative and biosorption potential, motility and chemotactic behavior, as well as physicochemical nature of the isolate was determined in the presence and absence of zinc. Optimum growth was observed at pH 7, 37 °C and at 1 mM phosphate. Minimum inhibitory concentration was found to be 20 mM for Zn2+ whereas metal resistance pattern for other heavy metal ions was Cu2+ (30 mM) > Ni2+ (25 mM) > Cr6+ (18 mM) > Pb2+ (12 mM) > Co2+ (8 mM) > Cd2+ (5 mM) > Hg2+ (3 mM). EC50 was found to be 3.4 mM Zn2+. Biosorption experiments in the presence of metal, DCCD and DNP in both live and killed cells demonstrated metal uptake, intracellular Zn2+ accumulation and adsorption by ATP independent efflux system. It was also found to effectively remove 81 mg/L and 87 mg/L after a period of 4 and 8 days, respectively. In the presence of Zn2+, positive chemotaxis was demonstrated where Zn2+ acted as a chemoattractant. Zn2+ induced elevated levels of GR, SOD and POX, while APOX and CAT activity were inhibited by it. The appearance of 92 kDa protein band in B. altitudinis under Zn2+ stress confirmed the induction of metallothionein. FTIR analysis revealed the active role of amide, hydroxyl, and carboxyl groups in combating Zn2+ stress. SEM and EDS analysis revealed surface adsorption of Zn2+ ions by bacterial cells. The experimental data obeyed the Freundlich isotherm model and pseudo second-order kinetics, indicating it to be an efficient biosorbent for Zn2+. This study revealed the potential application of B. altitudinis MT422188 in the bioremediation of Zn2+ from polluted wastewaters.

First report of Bacillus altitudinis causing seed rot of pomegranate in China

Pomegranate (Punica granatum L.) is an economically important fruit of the Punicaceae family and widely cultivated in the world. In 2019 and 2020, pomegranate fruits showing rot symptoms initiating from seed were observed in different pomegranate-cultivated fields in Henan province of China. Bacteria as a potential pathogen were isolated from the rot tissue of pomegranate seeds, and its pathogenicity was confirmed. The bacterium was identified as Bacillus altitudinis based on morphology and sequence analysis on the 16S ribosomal DNA and the gyrB gene. This is the first report on Bacillus altitudinis causing seed rot of pomegranate.