Isolation of Bacillus subtilis strain SEM-2 from silkworm excrement and characterisation of its antagonistic effect against Fusarium spp.

2020 ◽  
Vol 66 (6) ◽  
pp. 401-412
Author(s):  
Qingrong Li ◽  
Sentai Liao ◽  
Junhong Wei ◽  
Dongxu Xing ◽  
Yang Xiao ◽  
...  

Fusarium wilt is a devastating soil-borne disease mainly caused by highly host-specific formae speciales of Fusarium spp. Antagonistic microorganisms play a very important role in Fusarium wilt control. Isolation of potential biocontrol strains has become increasingly important. Bacterial strain SEM-2 was isolated from the high-temperature stage of silkworm excrement composting. SEM-2 exhibited a considerable antagonistic effect against Fusarium graminearum mycelial growth and spore germination. The results of pot experiments suggested that SEM-2 has a better inhibitory effect on the early stage of disease occurrence. The green fluorescent protein labelled SEM-2 coated on the surface of tomato seeds colonised the roots of tomato plants in 15 days. Genome sequencing identified SEM-2 as a new strain of Bacillus subtilis, and genome annotation and analysis determined gene clusters related to the biosynthesis of antimicrobials, such as bacillaene, fengycin, bacillibactin, subtilosin A, surfactin, and bacilysin. Interestingly, liquid chromatography – quadrupole time-of-flight mass spectrometry revealed that metabolites in pathways associated with the synthesis of secondary metabolites and antibiotics were highly differentially expressed. These findings may help to explain the mode of action of B. subtilis SEM-2 against Fusarium spp.

2019 ◽  
Vol 65 (1) ◽  
pp. 45-58 ◽  
Author(s):  
Qingrong Li ◽  
Sentai Liao ◽  
Huyu Zhi ◽  
Dongxu Xing ◽  
Yang Xiao ◽  
...  

Fusarium wilt is a devastating soil-borne disease caused mainly by highly host-specific formae speciales of Fusarium oxysporum. Antagonistic microorganisms play a very important role in Fusarium wilt control, and the isolation of potential biocontrol strains is becoming more and more important. We isolated a bacterial strain (SEM-9) from the high-temperature stage of silkworm excrement composting, which had a marked ability to solubilize phosphorus, promote the growth and increase the yield of the small Chinese cabbage, and which also exhibited considerable antagonistic effect towards Fusarium sambucinum and other fungi. The result of physiological and biochemical analyses, as well as genome sequencing, showed that SEM-9 was a strain of Bacillus subtilis. Through genome annotation and analysis, it was found that SEM-9 contained genes related to the regulation of biofilm formation, which may play an important role in colonization, and gene clusters encoding the biosynthesis of antimicrobials, such as surfactin, bacilysin, fengycin, and subtilosin-A. The production of such antifungal compounds may constitute the basis of the mode-of-action of SEM-9 against Fusarium spp. These data suggested that the SEM-9 strain has potential as both a biofertilizer and a biocontrol agent, with the potential to manage Fusarium wilt disease in crops.


2002 ◽  
Vol 184 (20) ◽  
pp. 5545-5553 ◽  
Author(s):  
Shigeo Hosoya ◽  
Kei Asai ◽  
Naotake Ogasawara ◽  
Michio Takeuchi ◽  
Tsutomu Sato

ABSTRACT In the course of a Bacillus subtilis functional genomics project which involved screening for sporulation genes, we identified an open reading frame, yaaT, whose disruptant exhibits a sporulation defect. Twenty-four hours after the initiation of sporulation, most cells of the yaaT mutant exhibited stage 0 of sporulation, indicating that the yaaT mutation blocks sporulation at an early stage. Furthermore, the mutation in yaaT led to a significant decrease in transcription from a promoter controlled by Spo0A, a key response regulator required for the initiation of sporulation. However, neither the level of transcription of spo0A, the activity of σH, which transcribes spo0A, nor the amount of Spo0A protein was severely affected by the mutation in yaaT. Bypassing the phosphorelay by introducing an spo0A mutation (sof-1) into the yaaT mutant suppressed the sporulation defect, suggesting that the yaaT mutation interferes with the phosphorelay process comprising Spo0F, Spo0B, and histidine kinases. We also observed that mutation of spo0E, which encodes the phosphatase that dephosphorylates Spo0A-P, suppressed the sporulation defect in the yaaT mutant. These results strongly suggest that yaaT plays a significant role in the transduction of signals to the phosphorelay for initiation of sporulation. Micrographs indicated that YaaT-green fluorescent protein localizes to the peripheral membrane, as well as to the septum, during sporulation.


Author(s):  
Mohammad Reza Sarjoughian ◽  
Fereshte Rahmani ◽  
Shamsozoha Abolmaali ◽  
Shakiba Darvish Alipour Astaneh

Background and Objectives: The great potential of bacteriophage for removing pathogen bacteria via targeting the cell wall is highly concerned. With a priority for overcoming drug-resistance, we screened against endolysins targeting Gram-negative bacteria to introduce a new antibacterial agent. This study was aimed to identify endolysins from the lysogenic phage of the Siphoviridea family in Bacillus subtilis. Materials and Methods: The Bacillus subtilis strain DDBCC46 was isolated from a preliminary antibacterial screening program. The endolysin (s) was extracted, concentrated with ammonium sulfate saturation, and their activity evaluated against the indicator bacteria. The phage particles were extracted from the bacteria using the minimum inhibition concentration of mitomycin C, followed by testing the phage inhibitory effect on the growth of indicator bacteria. The NCBI, Virus-Host DB, and EXPASY databases were used to obtain and confirm the sequences of the genes encoding PG hydrolases in Siphoviridea phages hosted in B. subtilis. Results: An 816 bp gene encoding an endolysin enzyme, was approved in the B. subtilis DDBCC 46, with specific primers of Bacillus phage SPP1. The purified-endolysin indicated antibacterial activity against Klebsiella pneumoniae, Salmonella typhimurium, Proteus (sp), and Escherichia coli. SDS-PAGE profiling followed by silica gel purification, led to introduceLys4630 as a therapeutic product and food preservative. Conclusion: lys4630 showed antibacterial effects on the common Gram-negative pathogens in clinics and food industries; E. coli, P. aeruginosa and Salmonella (sp).


2018 ◽  
Vol 33 (1) ◽  
pp. 9-18 ◽  
Author(s):  
Svetlana Milijasevic-Marcic ◽  
Vesna Todorovic ◽  
Olja Stanojevic ◽  
Tanja Beric ◽  
Slavisa Stankovic ◽  
...  

In vitro antagonistic potential of eleven isolates of Bacillus spp. against two phytopathogenic bacteria and one fungus was tested in order to identify potential biocontrol agents in vegetable crops. The Bacillus spp. isolates demonstrated different levels of antagonistic effect against the tested pathogenic microorganisms. Data in the study proved Xanthomonas vesicatoria to be more sensitive to Bacillus spp. strains than Clavibacter michiganensis subsp. michiganensis. Ten B. subtilis strains induced growth inhibition of X. vesicatoria, while a strain of B. pumilus did not affect the growth of that bacterium. The largest inhibition zones against X. vesicatoria were induced by strains B-319, B-325 and B-358. The pathogenic strain C. michiganensis subsp. michiganensis was most inhibited by two B. subtilis strains (B-338 and B-348) with mean inhibition zone diameters of up to 20 mm. B. subtilis strain B-319 which was the best in inhibiting X. vesicatoria, showed the lowest inhibitory effect on C. michiganensis subsp. michiganensis. The largest growth inhibition percentage of Verticillium sp. (PGI approximately 70%) was induced by B. subtilis strains B-310 and B-322. The other B. subtilis strains showed PGI values ranging from 45% to 68%, while B. pumilus strain B-335 had the least antagonistic potential (PGI =34.43%) against the pathogen. This study identified at least one suitable biocontrol candidate, B. subtilis strain B-358, as effective in vitro against all three vegetable pathogens.


Author(s):  
M. D. Tsulukidze ◽  
Z. Sh. Lomtatidze ◽  
Q. H. Mamulashvili

Aim. To study the antagonistic activity of actinomycetes isolated from soils adjacent to Kumisi Lake (Georgia). Methods. It was used 12 samples of actinomycetes isolated from the soils adjacent to Kumisi Lake. The antibiotic activity of actinomycetes was determined using test strains: Escherichia coli К-12 (B-3254), Bacillus subtilis 26 D, Xanthomonas campestris В-1459, Rhizobium radiobacter C58, Pectobacterium carotovorum EC1by the method of agar blocks. Results. It was revealed that actinomycete samples isolated from soils adjacent to Lake Kumisi shown the most significant inhibitory effect on test strains: Escherichia coli B-3254, Rhizobium radiobacter C58 and Xanthomonas campestris В-1459. The most resistant test-strains, to the studied actinomycetes, were Bacillus subtilis 26 D and Pectobacterium carotovorum EC1. Conclusion. Isolated strain 301 of actinomycetes shown a significant antagonistic effect on all test strains and could be potential for further studies spectrum and properties of antibiotic compounds.


Agriculture ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1273
Author(s):  
Sajid Iqbal ◽  
Nimat Ullah ◽  
Hussnain Ahmed Janjua

Recently, crop management has involved excessive use of chemical fertilizers and pesticides, compromising public health and environmental integrity. Rhizobacteria, which can enhance plant growth and protect plants from phytopathogen, are eco-friendly and have been attracting increasing attention. In the current study, Bacillus subtilis RS10 isolated from the rhizosphere region of Cynodon dactylon, inhibited the growth of indicator strains and exhibited in vitro plant growth-promoting traits. A whole-genome analysis identified numerous biosynthetic gene clusters encoding antibacterial and antifungal metabolites including bacillibactin, bogorol A, fengycin, bacteriocin, type III polyketides (PKs), and bacilysin. The plant growth-promoting conferring genes involved in nitrogen metabolism, phosphate solubilization, hydrogen sulfide, phytohormones, siderophore biosynthesis, chemotaxis and motility, plant root colonization, lytic enzymes, and biofilm formation were determined. Furthermore, genes associated with abiotic stresses such as high salinity and osmotic stress were identified. A comparative genome analysis indicated open pan-genome and the strain was identified as a novel sequence type (ST-176). In addition, several horizontal gene transfer events were found which putatively play a vital role in the evolution and new functionalities of a strain. In conclusion, the current study demonstrates the potential of RS10 antagonism against important pathogens and plant growth promotion, highlighting its application in sustainable agriculture.


2019 ◽  
Vol 34 (2) ◽  
pp. 97-102
Author(s):  
Ivana Potocnik ◽  
Svetlana Milijasevic-Marcic ◽  
Olja Stanojevic ◽  
Tanja Beric ◽  
Slavisa Stankovic ◽  
...  

The study aimed to isolate potential biocontrol agents from mushroom substrate that could serve as an alternative to toxic chemicals commonly used for disease control in mushroom production. The antagonistic potential of ten native Bacillus subtilis strains against the causal agents of green mould disease of oyster mushroom, Trichoderma pleuroti and Trichoderma pleuroticola, was evaluated. The antagonistic potential of Bacillus spp. strains was quantified in vitro based on dual cultivation with the pathogen. Growth inhibition of T. pleuroti ranged from 54.44% to 62.22% and no significant differences in antagonistic activity were found between the tested B. subtilis strains. Inhibition of T. pleuroticola was slightly higher, ranging from 55.56% to 69.62% and B. subtilis strain B-358 induced the highest growth inhibition. This research confirmed mushroom substrate to be a good source of antagonistic microorganisms with potentials for use in biological control of green mould in oyster mushroom production.


2021 ◽  
Vol 10 (4) ◽  
Author(s):  
Claudia Y. Muñoz ◽  
Anne de Jong ◽  
Oscar P. Kuipers

ABSTRACT Four strains isolated from tomato and lettuce phyllosphere were sequenced in order to investigate the presence of novel antimicrobial gene clusters and to get a better understanding of plant microbe interactions. These strains comprise two Bacillus strains, one Paenibacillus strain, and one Acinetobacter strain.


BioResources ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. 2789-2803
Author(s):  
Congxun Huang ◽  
Lihai Wang ◽  
Guoqi Xu ◽  
Pengwei Zhao ◽  
Chuhong Wang

Wood sapstain is a serious problem caused by the wood stain fungus, and it has a great influence on the international wood industry. The utilization of biological methods has good prospects for wood conservation. The objective of this study was to systematically estimate the antagonistic effect and influencing factors of Bacillus subtilis against wood stain fungus by using meta-analysis of literature data. Through report retrieval, a total of 992 references on B. subtilis related to wood were obtained. After strict screening, 163 data items from 7 articles were integrated. Estimated by the random-effects model, the combined effect Odds Ratio of the overall antagonistic effect was 0.15 (95% confidence interval [0.06, 0.34]). The results showed that B. subtilis could produce significant antagonistic effects against wood stain fungi. The inhibitory effect of wood stain fungi was affected by the strains of B. subtilis, species of wood stain fungi, the B. subtilis dosage, the type of mixed reagent, and the amount of mixed reagent on different wood stain fungi. The results of this study may provide a reference for biological control experiments, field tests, and practical applications of wood conservation.


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