Abstract: One hundred thirty-four putative Bacillus isolates were recovered from soybean rhizosphere soils of Nimar region to select effective zinc solubilizers for increased assimilation of zinc (Zn) in soybean seeds. These isolates were screened in vitro for zinc-solubilization ability on Tris-minimal agar medium supplemented separately with 0.1% zinc in the form of zinc oxide, zinc phosphate, and zinc carbonate. Of all, 9 isolates and a reference Bacillus cereus ATCC 13061 were characterized and identified as Bacillus species based on Gram-positive reaction, endospore-forming cells, and the presence of iso-C₁₅:₀ and anteiso-C₁₅:₀ as predominant fatty acids. On plate assay, two isolates KHBD-6 and KHBAR-1 showed a greater diameter of solubilization halo and colony diameter on all the three zinc compounds. The isolates KHBD-6, KHBAR-1, BDSD-2-2C, and KHTH-4-1 and the reference strain ATCC 13061 had higher soluble zinc concentration in liquid medium supplemented with zinc phosphate and zinc carbonate compounds as compared with the other isolates and uninoculated control. Evaluation under microcosm conditions showed that inoculation of isolates KHBD-6 (57.34 μg/g), KHBAR-1 (55.67 μg/g), and strain ATCC 13061 (53.10 μg/g) significantly increased the Zn concentration in soybean seeds as compared with the other isolates and uninoculated control (47.14 μg/g). This study suggests the occurrence of zinc-solubilizing Bacillus in soils of Nimar region and isolates KHBD-6 and KHBAR-1 were found to be promising zinc solubilizers for increased assimilation of Zn in soybean seeds.

Abstract: About 200 Bacillus isolates were isolated from tomato and potato rhizosphere and examined for their antagonistic activities against Ralstonia solanacearum T-91, the causal agent of tomato bacterial wilt (TBW), in vitro and in vivo . Four strains, AM1, D16, D29 and H8, have shown high potential of antagonistic activity against the pathogen in laboratory and greenhouse experiments. In greenhouse, 81.1 to 89.0% reduction of disease incidence of TBW was recorded in treated tomato plants with 4 isolates, which also significantly (p > 0.05) increased plant height by 22.7 to 43.7% and dry weight by 47.93 to 91.55% compared with non-treated control. 16SrRNA gene sequence, the biochemical and physiological tests and fatty acid methyl esters analysis assigned strains AM1 and D29 as Bacillus amyloliquefaciens , while strains D16 and H8 as Bacillus subtilis and B. methylotrophicus , respectively. In addition, the 4 strains showed ability to inhibit growth of the three soil-borne fungi, produce indole-3- acetic acid, siderophores and also with exception of strain D16, the other 3 strains were capable of solubilizing phosphate. Therefore, these results suggest that out of 200 isolates, Bacillus stains AM1, D16, D29 and H8 support good antagonistic activity and could be applied as biocontrol agents against TBW under greenhouse conditions beside their potential to promote tomato plants growth. Key words: Tomato, Ralstonia solanacearum , Bacillus spp, biological control, plant growth promotion activities

Abstract: Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plantgrowth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 µM auxin per µg of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.

Abstract: A large number of different microorganisms are commonly found in the soil including bacteria, fungi, ac- tinomycetes, protozoa and algae of these bacteria are by far the most common type of soil microorganism possibly because they can grow rapidly and have the ability to utilize a wide range of substances as either carbon or nitrogen sources. Use of naturally occurring, free living bacterial species, which can protect and promote plant growth by colonizing and multiplying along the surface of the root and/or root cortex. In our present investigation was to study the plant growth promoting (PGP) activities and chromium reducing Bacillus sp. from rice fields of in and around Erode district. From 25 soil samples 63 different Bacllius sp. were isolated. Among the 63, eight Bacillus sp. (BA1 to BA 8) possess effective PGP activities. In eight different Bacillus isolates particularly (BA1, BA3, BA4 and BA6) exhibited maximum plant growth promoting and chro- mium reducing activities. In addition to these traits, plant growth promoting bacterial isolates must be rhizospheric competent, able to survive and colonize in the rhizospheric soil.

Abstract: The present study was undertaken to find out the potential of indigenously isolated Bacillus spp. in controlling virulent luminous Vibrio harveyi infection in shrimp in vitro and in vivo. Luminescent bacteria were isolated from shrimp farms of east coast of India. It is biochemically and genetically identified as Vibrio harveyi. Two isolates of Vibrio harveyi obtained from infected shrimps were used for pathogenicity studies in juvenile shrimps and the LD50 values for the above isolates ranged between 3.7 × 10 6 to 8.9 × 10 7 CFU/ml. Indigenously isolated and identified Bacillus spp. exhibiting antagonism were used for controlling the Vibrio infection both in vivo and in vitro. In vivo antagonistic activity was studied with three selected Bacillus isolates. The same had reduced the mortality of Penaeus monodon juveniles when challenged with V. harveyi with an RPS of 87.5 - 50%. This is without affecting the multiplication of the pathogen in water. Present study found that indigenously isolated antagonistic Bacillus sp. could be used as effective biocontrol agents against pathogenic luminescent V. harveyi.

Abstract: Aims: The aim of this study was to identify Bacillus isolates capable of degrading sodium caseinate and subsequently to generate bioactive peptides with antimicrobial activity. Methods and results: Sodium caseinate (2·5% w/v) was inoculated separately with 16 Bacillus isolates and allowed to ferment overnight. Protein breakdown in the fermentates was analysed using gel permeation-HPLC (GP-HPLC) and screened for peptides (<3-kDa) with MALDI-TOF mass spectrometry. Caseicin A (IKHQGLPQE) and caseicin B (VLNENLLR), two previously characterized antimicrobial peptides, were identified in the fermentates of both Bacillus cereus and Bacillus thuringiensis isolates. The caseicin peptides were subsequently purified by RP-HPLC and antimicrobial assays indicated that the peptides maintained the previously identified inhibitory activity against the infant formula pathogen Cronobacter sakazakii. Conclusions: We report a new method using Bacillus sp. to generate two previously characterized antimicrobial peptides from casein. Significance and impact of the study: This study highlights the potential to exploit Bacillus sp. or the enzymes they produce for the generation of bioactive antimicrobial peptides from bovine casein.

Abstract: Among 21 different nitrogen-fixing Bacillus isolates obtained from soil, 6 exhibited co-production of indole-3-acetic acid (IAA) and bacteriocin-like inhibitory substance (BLIS). One of these isolates was identified as Bacillus pumilus based on its phenotypical and biochemical properties as well as its 16S rRNA gene sequence; this isolate yielded 5.6 mg/biomass IAA co-production along with a potent BLIS production. The antimicrobial activity and proteinaceous nature of this BLIS were confirmed, and it was identified as a novel <25 kDa UV-stable peptide by SDS-PAGE. The partially purified BLIS was active against a broad range of food-borne, hospital pathogenic and spoilage bacteria, including Listeria monocytogenes, Bacillus cereus, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant Enterococcus (VRE). In addition, it showed ideal physico-chemical properties in terms of ultraviolet and thermal resistance, water solubility, and stability against acid/alkali (pH 2.0–9.0) treatment. Finally, the effects of different variables, such as physical parameters and key media components, on determining the optimal condition of BLIS and IAA production were investigated using the Taguchi method. The co-production of a UV-stable BLIS—the first report for this isolate—and IAA demonstrate the prospective multilateral application of this Bacillus pumilus isolate as a biofertilizer and probiotic.

Abstract: Bacillus isolates were analyzed for alkaline protease production on casein containing agar plates and identified by clear zones of casein hydrolysis around colonies. Two gram positive Bacillus isolates S5 and C3 which showed the best enzyme production were studied for purification by precipitation and dialysis. The intracellular alkaline protease enzyme was purified about 4 fold with a yield of 9.8% for isolate S5 and about 2 fold with a yield of 18% for isolate C3. The pH and temperature optima of partially purified alkaline protease were determined to be 9 and 40°C, respectively. Such alkaliphilic proteases from these isolates have a great economical and environmental impact to alleviate the pollution problem created by leather and other industries. Soil and water pollution from hazardous lime, sodium Materials: All the bacteriological media components were sulphide, solvents, etc. the end-products of the pre- product of Hi-Media, India. All other chemicals were of tanning stages in the leather processing industries is analytical grade. Bacillus isolates were procured from the increasing at a high pace. Proteases can be used for the gene pool of the Dept. of Biochemistry, G.B.P.U.A.T. replacement of chemicals involved during tanning Pantnagar. procedures to solve this problem. Of particular importance are the proteases with activity at alkaline pH and high Methods: temperatures (1, 2). Cultivation and Screening of Positive Strains: Bacillus Protease is responsible for cell growth and isolates (procured from the gene pool of the Dept. of differentiation and is found in all living organisms. Out of Biochemistry, G.B.P.U.A.T. Pantnagar) plated onto 'Skim many commercial protease producers only few bacterial Milk Agar' plates (8) containing peptone (0.5% w/v), NaCl strains are recognized. Bacillus sp. dominate the (0.5% w/v), agar (1.5% w/v) and skim milk (10% w/v). industrial sector being active producers of extracellular Plates were incubated at 37°C for 24 hours. A clear zone proteases. Alkaline proteases alone account for 20% of of skim milk hydrolysis gave an indication of protease the world enzyme market with their predominant use in producing organisms. leather processing and detergent industries (3). It has been proved that alkaline protease from Bacillus can be Enzyme Production: Production of protease from used as a dehairing agent, indicating elastolytic, Bacillus strains was carried out in a 'Protease keratinolytic activities and also a low hydrolytic collagen Specific Medium' (6) containing glucose (0.5% w/v), activity (4-7). peptone (0.75% w/v) and salt solution (5% v/v) i.e. In the present study, it could be attemped to isolate MgSO .7H O (0.5% w/v), KH PO (0.5% w/v); and bacteria from soil and optimize it for protease production FeSO .7H O (0.01% w/v) maintained at 37°C for 48 hours by a promising strain. Different bacterial strains were in a shaker incubator (120 rpm). The pH of the isolated from soil and screened for their ability to produce medium was adjusted. The contents were then centrifuged protease and two potential producers were obtained. at 7, 000 rpm for 20 min. at 4°C and the cell-free Here, we report the purification and characterization of the supernatant was used for determining extracellular extracellular alkaline protease from B. strains. protease activity.

Abstract: The aim of the investigation was to study the hydrolytic enzymes viz., chitinase, protease, , β-1, 3 glucanase and cellulase from the isolates of Bacillus sp. (twenty eight) which isolated from tomato rhizospheric soil in IIVR farm (DPNSB-1 to 7), IIHR farm (DPNSB-8 to 15), IARI farm (DPNSB-16 to 20) and farm of APHU (DPNSB-21 to 28). Among the strains, IARI isolate of DPNSB-18 exhibited the highest chitinase activity (4.65 IU/ml), IIHR isolate of DPNSB-15 produce highest protease activity (0.79 IU/ml), maximum , β-1, 3 glucanase production was noted in Bacillus strains viz., DPNSB-14 (IIHR isolate), DPNSB-2 (IIVR isolate) and DPNSB-20 (IARI isolate), range from 0.24 IU/ml to 0.39 IU/ml, cellulase production was made by isolates of IIVR, DPNSB-3 (0.75 IU/ml) and DPNSB-1 (0.60 IU.ml) respectively.

Abstract: Bacillus species form an heterogeneous group of Gram-positive bacteria that include members that are disease-causing, biotechnologically-relevant, and can serve as biological research tools. A common feature of Bacillus species is their ability to survive in harsh environmental conditions by formation of resistant endospores. Genes encoding the universal stress protein (USP) domain confer cellular and organismal survival during unfavorable conditions such as nutrient depletion. As of February 2012, the genome sequences and a variety of functional annotations for at least 123 Bacillus isolates including 45 Bacillus cereus isolates were available in public domain bioinformatics resources. Additionally, the genome sequencing status of 10 of the B. cereus isolates were annotated as finished with each genome encoded 3 USP genes. The conservation of gene neighborhood of the 140 aa universal stress protein in the B. cereus genomes led to the identification of a predicted plasmid-encoded transcriptional unit that includes a USP gene and a sulfate uptake gene in the soil-inhabiting Bacillus megaterium. Gene neighborhood analysis combined with visual analytics of chemical ligand binding sites data provided knowledge-building biological insights on possible cellular functions of B. megaterium universal stress proteins. These functions include sulfate and potassium uptake, acid extrusion, cellular energy-level sensing, survival in high oxygen conditions and acetate utilization. Of particular interest was a two-gene transcriptional unit that consisted of genes for a universal stress protein and a sirtuin Sir2 (deacetylase enzyme for NAD+-dependent acetate utilization). The predicted transcriptional units for stress responsive inorganic sulfate uptake and acetate utilization could explain biological mechanisms for survival of soil-inhabiting Bacillus species in sulfate and acetate limiting conditions. Considering the key role of sirtuins in mammalian physiology additional research on the USP-Sir2 transcriptional unit of B. megaterium could help explain mammalian acetate metabolism in glucose-limiting conditions such as caloric restriction. Finally, the deep-rooted position of B. megaterium in the phylogeny of Bacillus species makes the investigation of the functional coupling acetate utilization and stress response compelling.

Abstract: Native isolates of Pseudomonas and Bacillus collected from the rhizosphere of different horticultural crops were tested for their efficacy against root-knot nematode, Meloidogyne incognita infesting black pepper. All the native isolates of Pseudomonas and Bacillus tested were shown to increase the germination per cent and enhanced the vigour index of rice under roll towel method. Under in vitro studies, greatest reduction in nematode egg hatching and highest juvenile mortality was observed in the culture filtrate of consortia of Pseudomonas isolate, Pf 123 and Bacillus isolate, Bs 214 at its 100 per cent concentration. Under glass house and field conditions, plants treated with the consortia, Pf 123 + Bs 214 significantly enhanced the plant growth, yield and reduced nematode infestation both in soil and root.

Abstract: 1 * In this research, antagonistic activities of 30 Bacillus species isolated from various fish samples were studied. Isolated Bacillus species were analyzed using the agar diffusion method in terms of their general inhibition effects against some food pathogen/ contaminant bacteria and lactic acid bacteria isolated from the fish intestinal tracts. Some of the strains exhibited antimicrobial activity against Bacillus subtilis, Pseudomonas fluorescens , Lactobacillus coryneformis, Lactobacillus plantarum and Lactobacillus xylosus. Furthermore, the inhibitory effects of the lactic acid bacteria on the Bacillus isolates were analyzed by using the same method.

Abstract: Plant diseases need to be controlled to maintain the quality and abundance of food, feed, and fiber produced by growers around the world. Different approaches may be used to Control plant diseases, Such as chemical fertilizers and pesticides. The environmental pollution caused by excessive use and misuse of agrochemicals some pest management have focused their efforts on developing alternative inputs to synthetic chemicals for controlling pests and diseases. Among these, alternatives are those referred to as biological control. Members of the U.S. National Research Council took into account modern biotechnological developments and referred to biological control as the use of natural or modified organisms, genes, or gene products, to reduce the effects of undesirable organisms and to favor desirable organisms such as crops, beneficial insects, and microorganisms”, but this definition spurred much subsequent debate and it was frequently considered too broad by many’ scientists who worked in the field (US Congress, 1995). The use of a grampositive Bacillus species as a biocontrol agent is relatively rare, and has received less intensive study than the use of gram-negative bacteria. The antagonists studied have been mainly Bacillus subtilis and occasionally B. megaterium, B. cereus, B. pumilus, and B. polymyxa (Utkhede, 1984). As Bacillus spp. have the characteristics of, being widely distributed in nearly all agricultural soils and in other environments, having high thermal tolerance, showing rapid growth in liquid culture, and readily form resistant spores. Moreover, they are considered safe biological agents, and their potential as Bio-control agents is considered to be high. However, the evaluation of bacteria has focused primarily on disease suppression (Siala and Gray, 1974). Bacillus spp. can be used as biological control agent for bacteria and fungal diseases like gray mold, powdery mildews, early and late blight, bacterial spot and walnut blight through production of antimicrobial proteins namely bacteriocin, chitinase, glucanase etc and antibiotics as well as antifungal synthesized by secondary metabolism pathways.

Abstract: Reporter bacteria are beneficial for the rapid and sensitive screening of cultures producing peptide antibiotics, which can be an addition or alternative to the established antibiotics. This study was carried out to validate the usability of specific reporter strains for the target mediated identification of antibiotics produced by native Bacillus spp. isolated from different food sources. During preliminary classification, cell wall stress causing Bacillus isolates were screened by using reporter strain Bacillus subtilis BSF2470. The isolates which induced cell wall stress were further characterized for their specific mode of action by using other B. subtilis reporter strains (TMB 488, TMB 299 and TMB 279). The isolate B. licheniformis N12 was found to produce bacitracin confirmed by the response to reporter strain B. subtilis TMB 279 and by putative identification of bacitracin biosynthetic loci. The other isolate B. subtilis EC1 also induced B. subtilis TMB 279, but does not possess the bacitracin gene cluster indicating that it can be a novel, bacitracin like antibiotic. The different but related subsets of peptide antibiotics that bind the pyrophosphate moiety of the lipid carrier of cell wall biosynthesis can be identified using this whole cell based reporter strains.

Abstract: Aims Pigmented Bacillus spp. with probiotic properties have been isolated. In the yellow-/orange-coloured strains, the carotenoid pigments present have been characterized. In contrast, the carotenoids present in the Bacillus isolates coloured red await identification. The present article reports progress on the elucidation of the pigment biosynthetic pathway in these red-pigmented Bacillus firmus strains. Methods and Results A combination of UV/Vis, chromatographic and mass spectrometry (MS) has revealed the properties of the predominant pigment and the end-point carotenoid of the pathway to be methyl 4,4′-diapolycopene-dioate after transmethylation. The diglycosyl ester of 4,4′-diapolycopene-dioate persists in vivo prior to chemical treatment. Different mutants and inhibitor treatment were employed to establish the C30 biosynthesis pathway with all precursors and intermediates to 4,4′-diapolycopene-dioate detected, which include 4,4′-diapophytene and all desaturation intermediates to 4,4′-diapolycopene and 4,4′-diapolycopene-dialdehyde. To cultures synthesizing the 4,4′-diapolycopene-dioate derivative and those in which its formation was inhibited, oxidative stress was induced by peroxide treatment. Conditions that decreased the growth rate of the pigmented cells by only 30% caused a complete growth inhibition of the culture devoid of the 4,4′-diapolycopene-dioate derivative. Conclusion This finding demonstrates the diversity of C30 carotenoid biosynthesis in Bacillus species and the antioxidative function of the 4,4′-diapolycopene-dioate derivative in B. firmus cells. Significance and Impact of the Study It could be shown that the C30 4,4′-diapolycopene-dioate derivatives protect pigmented B. firmus from peroxidative reactions. Under oxidative conditions, this can be an ecological advantage over nonpigmented (=noncarotenogenic) strains that are equally abundant.