scholarly journals Chitinolytic Bacillus subtilis Ege-B-1.19 as a biocontrol agent against mycotoxigenic and phytopathogenic fungi

2018 ◽  
Vol 44 (3) ◽  
pp. 323-331
Author(s):  
Ozlem Oztopuz ◽  
Nermin Sarigul ◽  
Fakhra Liaqat ◽  
Ro-Dong Park ◽  
Rengin Eltem
Keyword(s):  
Molecular Weight ◽  
Bacillus Subtilis ◽  
Biocontrol Agent ◽  
Pathogenic Fungi ◽  
Phytopathogenic Fungi ◽  
Aspergillus Ochraceus ◽  
Lytic Enzyme ◽  
Rrna Gene ◽  
Lytic Enzymes ◽  
16S Rrna Gene Analysis

Abstract Background Biological control of pathogenic fungi is a possible alternate to the chemical control, which is harmful to humans and environment. Soil-borne Bacillus strains can be potential biocontrol agents and a source of lytic enzymes. Aim This study aimed to examine biocontrol potential and lytic enzyme activities of a soil isolate Bacillus subtilis Ege-B-1.19. Materials and methods Strain was identified by biochemical and 16S rRNA gene analysis and its biocontrol activity was investigated against Aspergillus niger EGE-K-213, Aspergillus foetidus EGE-K-211, Aspergillus ochraceus EGE-K-217, Fusarium solani KCTC6328, Rhizoctonia solani KACC40111 and Colletotrichum gloeosporioides KACC40689. Chitinase, chitosanase, N-acetyl-β-hexosaminidase and protease activities of B. subtilis Ege-B-1.19 were also determined. Chitosanase was purified using Sephadex G-150 column and its molecular weight was determined by SDS-PAGE. Chitooligosaccharides production using chitosanase was carried out and analysed by TLC and HPLC. Results Results depicted that B. subtilis Ege-B-1.19 has shown inhibitory effects against all the test fungi. Chitinase, chitosanase, N-acetyl-β-hexosaminidase and protease activities were determined as 2.7 U mL−1, 7.2 U mL−1, 6.2 U mL−1 and 12.2 U mL−1, respectively. Molecular weight of purified chitosanase was 44 kDa. Chitosanase hydrolysed chitosan to glucosamine (GlcN), dimers (GlcN)2 and trimers (GlcN)3. Conclusion Bacillus subtilis Ege-B-1.19 can be anticipated as useful biocontrol agent and its chitosanase can be utilized for enzymatic synthesis of chitooligosaccharides.

scholarly journals Studies of some genes of Bacillus subtilis MN99 local strains against plant diseases

2021 ◽  
Vol 33 (2) ◽  
pp. 38-44
Author(s):  
Ariunaa Saraadanbazar ◽  
Byambasuren Mijidsuren ◽  
Battur Banzragch
Keyword(s):  
Bacillus Subtilis ◽  
Local Strain ◽  
Pathogenic Fungi ◽  
Plant Diseases ◽  
Likelihood Method ◽  
Partial Sequence ◽  
Rrna Gene ◽  
Multiple Sequence ◽  
Local Strains ◽  
Isolation And Characterization

The objective of this study was the isolation and characterization of Bacillus subtilis local strains from the soil in Mongolia. These local strains of B. subtilis are showed to have high antagonistic activities against some plant pathogenic fungi and bacteria. Six strains of B. subtilis were isolated and characterized morphologically, physiologically and biochemically according the Bergey’s Manual of Systematic Bacteriology. In order to identify species of the isolated strains, we amplified and sequenced 16S rRNA gene, essential funtinal genes bmyB, spoVG and srfAA, which are related to antagonistic activity of these strains. The sequences were aligned using CLASTALW multiple sequence alignment tool. Phylogenetic tree was drawn according to Maximum likelihood”method and “Tamura-Nei” model using “MEGA-X version 10.2.6 program. Among all isolates of B. subtilis MN99 and 7/24 strains had higher antagonist activity against plant diseases. According to partial sequence of srfAA (620bp) gene of MN99, the local strain belongs to B. subtilis and partial sequence of bmyB (370bp), spoVG (22bp) gene of MN7/24 strain showed that the it belongs to B. atrophaeus species. All local strains of B. subtilis had bacillomycin synthesis gene, and B. subtilis MN99 strain had only surfactine synthesis gene, while did not have spore formation and hemolysis gene SpoVG.

scholarly journals The Use of Ascophyllum nodosum and Bacillus subtilis C-3102 in the Management of Canine Chronic Inflammatory Enteropathy: A Pilot Study

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3417
Author(s):  
Marco Isidori ◽  
Fabrizio Rueca ◽  
Francesca Romana Massacci ◽  
Manuela Diaferia ◽  
Andrea Giontella ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Activity Index ◽  
Disease Activity Index ◽  
Ascophyllum Nodosum ◽  
Rrna Gene ◽  
16S Rrna Gene Analysis ◽  
Positive Effects ◽  
Inflammatory Bowel ◽  
Bacillus Genus ◽  
Compositional Variations

The aim was to assess the effects of Ascophyllum nodosum (AN) with/without Bacillus subtilis C-3102 as alternative treatments for Chronic Inflammatory Enteropathy (CIE) of dogs. Fourteen CIE patients, which had received the same control (CTR) diet, were enrolled to serially receive three diets: (1) hydrolysed protein (HP) diet; (2) 4.0% AN supplemented HP (HPA) food, (3) HPA diet fortified with 125 billion B. subtilis C-3102 spores/10 kg body weight (HPAB diet). Clinical outcome was assessed by Canine Inflammatory Bowel Disease Activity Index (CIBDAI), whereas gut microbiota compositional variations were investigated via 16S rRNA gene analysis, and faecal fermentation end-products by liquid chromatography. Higher abundances of the Ruminococcaceae and Rikenellaceae families were shown in HPA relative to CTR treatment, with Bacillus genus being differentially abundant on HPAB diet. Concentrations of acetate were higher (p < 0.05) in dogs fed HPA compared to CTR diet, and amounts of isovalerate and isobutyrate were greater (p < 0.05) in HPA compared to HP food. A tendency for higher amounts of faecal butyrate was found for the HPAB treatment (p = 0.06). Comprehensively, while displaying potentially positive effects on faecal fermentations, the tested substances failed to improve CIBDAI scores and microbial richness in CIE dogs.

scholarly journals Potential Application of Rhizobacteria Isolated from the Central Highland of Vietnam as an Effective Biocontrol Agent of Robusta Coffee Nematodes and as a Bio-Fertilizer

Agronomy ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1887
Author(s):  
Dai Nam Nguyen ◽  
San-Lang Wang ◽  
Anh Dzung Nguyen ◽  
Manh Dung Doan ◽  
Dinh Minh Tran ◽  
...  
Keyword(s):  
Plant Growth ◽  
High Performance ◽  
Biocontrol Agent ◽  
Pathogenic Fungi ◽  
Rrna Gene ◽  
Inhibition Rate ◽  
Egg Hatching ◽  
Chromatography Analysis ◽  
Robusta Coffee ◽  
Central Highland

Robusta coffee is a major commercial crop in the Central Highland of Vietnam with high economic and export value. However, this crop is adversely affected by various pathogens, particularly nematodes. This study aimed to screen active anti-nematode rhizobacterial strains for sustainable coffee production. Among more than 200 isolates, the isolate TUN03 demonstrated efficient biocontrol with nearly 100% mortality of J2 coffee nematodes Meloidogyne spp. and 84% inhibition of nematode egg hatching. This active strain was identified as Pseudomonas aeruginosa TUN03 based on its 16S rRNA gene sequence and phylogenetic analysis. In greenhouse tests, the strain TUN03 significantly reduced the coffee nematode population in the rhizome-soil with an 83.23% inhibition rate and showed plant growth-promoting effects. Notably, this is the first report of the nematicidal effect of P. aeruginosa against coffee nematodes. This potent strain further showed an antifungal effect against various crop-pathogenic fungi and was found to be the most effective against Fusarium solani F04 (isolated from coffee roots) with a 70.51% inhibition rate. In addition, high-performance liquid chromatography analysis revealed that this bacterial strain also secretes plant growth regulators including indole acetic acid (IAA), gibberellic acid (GA3), kinetin, and zeatin in significant amounts of 100, 2700, 37, and 9.5 µg/mL, respectively. The data from this study suggest that P. aeruginosa TUN03 may be a potential biocontrol agent and biofertilizer for the sustainable production of Robusta coffee and other crops.

scholarly journals THE COMPOST, A SOURCE OF PLANT BENEFICIAL BACTERIA WITH BIOCONTROL POTENTIAL

2021 ◽  
Vol 10 (20) ◽  
pp. 32-38
Author(s):  
Oana-Alina Boiu-Sicuia ◽  
Vasilica Stan ◽  
Călina Petruța Cornea
Keyword(s):  
Plant Protection ◽  
Bacterial Load ◽  
Pathogenic Fungi ◽  
Phytopathogenic Fungi ◽  
Microbial Load ◽  
Bacterial Strains ◽  
Beneficial Bacteria ◽  
Lytic Enzymes ◽  
Common Activity ◽  
Biocontrol Potential

Recycling the sewage sludge from treatment plants is a common activity. The resulting compost is usually rich in plant nutrients and beneficial microorganisms. However, compost properties greatly differ depending on the nature of the fermented biomass and fermentation processes. The aim of this study was to analyze the microbial load of three different composts, in order to detect new bacterial strains with plant protection properties. Isolated bacteria were microbiologically characterized and evaluated for their potential to reduce soil-borne phytopathogenic fungi. Results showed a microbial load of approximately 106 CFU/g of compost. In the analyzed samples it was revealed that as bacterial load increases, the fungal amount decreases. Analyzing some newly isolated bacteria obtained from these composts, a good biocontrol potential against soil-borne pathogenic fungi was revealed. Some of the isolated bacterial strains revealed antifungal activity against Rhizoctonia solani and Sclerotinia sclerotiorum. These bacteria showed good colonization capacity and lytic enzymes production, correlated to antimicrobial activity. These compost-originated bacteria reveal high potential in pathogens inhibition. Therefore, the analyzed composts are recommended not only as soil fertility improvers, but also as potential suppressors of soil-borne pathogens. Results revealed these composts as source of plant beneficial bacteria with biological control potential.

scholarly journals A novel antifungal peptide purified from Bacillus subtilis strain EDR4

2016 ◽  
Vol 68 (2) ◽  
pp. 319-324
Author(s):  
Jingyuan Ji ◽  
Jie Yang ◽  
Lili Huang ◽  
Zhensheng Kang
Keyword(s):  
Bacillus Subtilis ◽  
Antifungal Activity ◽  
Biocontrol Agent ◽  
Pathogenic Fungi ◽  
Subtilis Strain ◽  
Antifungal Peptide ◽  
Inhibitory Effects ◽  
Canker Disease ◽  
Ph Values ◽  
Potential Biocontrol Agent

Bacillus subtilis strain EDR4 is a potential biocontrol agent against Valsa mali (Vm), the fungus causing apple canker disease. In this study, we identified and characterized the antifungal peptide produced by B. subtilis EDR4. Peptides were isolated by 30% ammonium sulfate precipitation and purified by column chromatography. A 4-kDa peptide exhibiting antifungal activity was obtained and designated as P6. The amino acid sequence of P6 was determined by liquid chromatography combined with tandem mass spectrometry. P6 showed inhibitory effects against eight different pathogenic plant fungi, and was stable (i.e., retained its biological activity) at temperatures as high as 121?C for 20 min and at pH values ranging from 3 to 11. Microscopic examination of Vm hyphae treated with P6 showed protoplasm release, and in vitrospore germination was also inhibited. These results suggest that P6 is the active substance responsible for the antifungal activity of B. subtilisEDR4 against Vmand that P6 may be effective in the biocontrol of Vmas well as other plant pathogenic fungi.

scholarly journals Lytic Enzymes of Aspergillus piperis as a Tool for Attacking Some Phytopathogenic Fungi In vitro with Special Reference to its Cytotoxicity

2021 ◽  
Author(s):  
Samah Abd El-Kader El-Debaiky ◽  
Anwer S.M. El-Badry
Keyword(s):  
Cell Line ◽  
Biological Control Agent ◽  
Control Agent ◽  
Phytopathogenic Fungi ◽  
Antagonistic Activity ◽  
Lytic Enzyme ◽  
Lytic Enzymes ◽  
Wi38 Cell ◽  
Ic50 Values

The antagonistic activity of Aspergillus piperis against Fusarium oxysporum f. sp. fabae (FOF) and Sclerotinia sclerotiorum were examined and showed multiple signs of hyphal interactions. Microscopic examination of contact regions among A. piperis and each pathogen revealed distinct enzymatic lysis of pathogenic hyphal cell walls. Therefore, it is important to estimate the lytic enzyme activity of A. piperis. Extracellular lytic enzymes are important offensive forces for A. piperis as a biological control agent. Chitinase, phospholipase, and protease recorded relatively high activity from a culture age of 10 days (82.3, 42.4, and 6.2 U/ml, respectively). Enzymatic persistence was measured at room temperature, recording relatively long periods, saving 54%, 46%, and 21% of their activity, respectively. The cytotoxicity of the crude culture filtrate of A. piperis was examined in MCF7 and WI38 human cell lines. The cell viability (IC50) value of the fungal filtrate was estimated after 24 h and 48 h. The results revealed that IC50 values against the MCF7 cell line were inoperative after 24 h and were recorded 80 μg/ml after 48 h. In contrast, IC50 values against the WI38 cell line were 85.69 and 69.8 μg/ml after 24 and 48 h, respectively.

scholarly journals Screening and identification of thermophilic cellulolytic bacteria isolated from sawdust compost

2021 ◽  
pp. 348-357
Author(s):  
D. T. H. Phuong ◽  
D. T. Tuyen ◽  
L. V. Thang
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Megaterium ◽  
Metabolic Pathways ◽  
Rrna Gene ◽  
Cellulolytic Bacteria ◽  
16S Rrna Gene Analysis ◽  
Wide Range ◽  
Commercial Enzymes ◽  
Screening And Identification ◽  
Physiological And Biochemical

Composting process mainly depends on the metabolic pathways of the microorganism and involves the activity of different enzymes. Thermophilic cellulase-producing bacteria isolated from sawdust compost were tested for formation of a visible zone around the colonies on the agar plates medium containing carboxymethyl cellulose at 50ºC. Screening of carboxymethyl cellulase producing isolates was further realized on the basis in liquid medium by DNS method. Among 29 isolates investigated, V1 and V11 strains exhibited maximum enzyme activity of 1.9 and 2.3 U/mL, respectively. These isolates were selected for morphological, physiological and biochemical studies and 16S rRNA gene analysis. They were found a Gram-positive, rod-shaped spore forming cells, which were identified as Bacillus megaterium (V1) and Bacillus subtilis (V11) based on cell morphology, nucleotide homology and phylogenetic analysis. The optimal temperature for activity of endoglucanases (CMCase) ranged from 35–45°C (strain V1) and 40– 50ºC (strain V11). Our findings showed that Bacillus megaterium (V1) and Bacillus subtilis (V11) cellulase demonstrate thermophilic characteristics within wide range of temperature and meets the requirements for commercial enzymes. 

scholarly journals Isolation of Burkholderia sp. HQB-1, A Promising Biocontrol Bacteria to Protect Banana Against Fusarium Wilt Through Phenazine-1-Carboxylic Acid Secretion

2020 ◽  
Vol 11 ◽  
Author(s):  
Zhizhou Xu ◽  
Mingyuan Wang ◽  
Jinpeng Du ◽  
Ting Huang ◽  
Jianfu Liu ◽  
...  
Keyword(s):  
Carboxylic Acid ◽  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Pathogenic Fungi ◽  
Phytopathogenic Fungi ◽  
Rrna Gene ◽  
Antifungal Compound ◽  
Gene Sequence Analysis ◽  
Banana Wilt ◽  
Physiological And Biochemical

Fusarium wilt is a devastating soil-borne fungal disease caused by Fusarium oxysporum f.sp. cubense (Foc). In recent years, some antifungal bacteria have been applied for the prevention and biocontrol of pathogenic fungi. In our study, a bacterial strain HQB-1, isolated from banana rhizosphere soil, was cultured for investigation. It showed broad-spectrum antifungal activities against representative phytopathogenic fungi including Fusarium oxysporum, Colletotrichum gloeosporioides, Botrytis cinerea, and Curvularia fallax. The strain HQB-1 was identified as Burkholderia sp. by morphological, physiological, and biochemical examinations, confirmed by 16S rRNA gene sequence analysis. Among the metabolites produced by the strain, we identified an antifungal compound which was identified phenazine-1-carboxylic acid (PCA) (C13H8N2O2) through ultraviolet, liquid chromatography quadrupole-time of flight mass spectrometer, and nuclear magnetic response. Furthermore, PCA exhibited the lowest minimum inhibitory concentration (MIC) against F. oxysporum (1.56 μg/ml) and yielded the highest MIC against C. gloeosporioides. Pot experiments showed that application of 5 μg/ml or more of PCA efficiently controlled banana wilt and promoted the growth of banana plants. These results suggested that Burkholderia sp. HQB-1, as an important microbial resource of PCA, could be a promising biological agent against wilt diseases and promoting banana growth.

scholarly journals In vitro potential of Paenibacillus alvei DZ-3 as a biocontrol agent against several phytopathogenic fungi

Biologija ◽  
2018 ◽  
Vol 64 (1) ◽  
Author(s):  
Natalija Atanasova-Pancevska ◽  
Dzoko Kungulovski
Keyword(s):  
Biocontrol Agent ◽  
Phytopathogenic Fungi ◽  
Diffusion Method ◽  
Rrna Gene ◽  
Dual Culture ◽  
Promising Alternative ◽  
Antagonistic Microorganisms ◽  
Paenibacillus Alvei ◽  
Fungal Phytopathogens

Fungal phytopathogens cause significant losses in many economically important crops and vegetables. One way to control these devastating pathogens is using higher doses of fungicides that not only increase the cost of production but also cause significant damage to the environment. Consequently, there is an increasing demand from consumers and officials to reduce the use of chemical pesticides. In this context, biological control through the use of natural antagonistic microorganisms has emerged as a promising alternative. The goal of this paper is to evaluate environmentally-friendly treatment for in vitro control of some fungal phytopathogens. In the present study, the bacterial strain DZ-3, which shows strong antifungal activity, was isolated from the samples of rotten apple compost from the composting plant in Resen, Macedonia, and identified as Paenibacillus alvei according to morphological and taxonomic characteristics and 16S rRNA gene sequence analysis. As test microorganisms, we used Botrytis cinerea FNSFCC 23, Fusarium oxysporum FNS- FCC 103, Plasmopara viticola FNS- FCC 65, Alternaria alternata FNS- FCC 624, but also Aspergilus ochraceus FNS- FCC 46, Aspergilus niger FNS- FCC 142, and Penicillium commune FNS- FCC 864. The effect of Paenibacillus alvei DZ-3 on growth of the tested fungi was evaluated by the dual culture technique and disc diffusion method. Clear inhibition zones were observed in all test microorganisms. The above-described results indicate that Paenibacillus alvei DZ-3 may have the potential as a biocontrol agent to control various phytopathogenic fungi.

scholarly journals Isolation of Cellulolytic Bacillus subtilis Strains from Agricultural Environments

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Yu-Kyoung Kim ◽  
Shin-Chan Lee ◽  
Young-Yun Cho ◽  
Hyun-Jeong Oh ◽  
Young Hwan Ko
Keyword(s):  
Bacillus Subtilis ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Biochemical Characterization ◽  
Soluble Sugars ◽  
Cellulolytic Enzyme ◽  
Rrna Gene ◽  
Human Society ◽  
Cellulolytic Bacteria ◽  
16S Rrna Gene Analysis

The bioconversion of cellulose and hemicellulose to soluble sugars is important for global stabilization and a sustainable human society. Here, hundreds of cellulolytic bacteria were screened and isolated from soil, compost, and animal waste slurry in Jeju Island, South Korea. Among the isolates, three strains, SL9-9, C5-16, and S52-2, showing higher potential for practical uses were purified on carboxymethyl cellulose (CMC) agar plates and identified as Bacillus subtilis strains by morphological, physiological, and biochemical characterization and 16S rRNA gene analysis. The production patterns of cellulose or hemicellulose-degrading enzymes were investigated during cell culture. All three isolated strains produced CMCase, Avicelase, β-glucosidase, and xylanase enzymes, which suggested synergic cellulolytic systems in Bacillus subtilis. The enzymes showing CMCase, Avicelase, and xylanase activities existed in cell-free culture supernatant, meanwhile β-glucosidase activity was detected in cell debris suggesting that three of the enzymes, including CMCase, Avicelase, and xylanase, were extracellular, and β-glucosidase was cell membrane bound. The three isolates, SL9-9, C5-16, and S52-2, were not the same strains, presenting slight differences in biochemical characteristics, 16S rRNA gene sequences, and cellulolytic enzyme activities.

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