Genome sequencing and analysis of Bacillus velezensis VJH504 reveal biocontrol mechanism against cucumber Fusarium wilt

Yang, Fan; Jiang, Huayan; Ma, Kai; Wang, Xin; Liang, Shen; Cai, Yuxin; Jing, Yancai; Tian, Baoming; Shi, Xuanjie

doi:10.3389/fmicb.2023.1279695

Cited by 8 publications

(5 citation statements)

References 85 publications

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“…B. velezensis could produce antimicrobial substances such as polyketides, lipopeptides, and peptides, which inhibit bacteria and fungi [21]. The secondary metabolites produced by B. velezensis involve bacillibactin, fengycin, bacilysin, surfactin, difficidin, and mersacidin, which exhibit broad antagonistic activities against pathogenic bacteria or fungi [22]. For example, anti-microbial substances such as bacillibactin chelated iron, fengycin altered cell wall permeability, bacilysin hindered glucosamine synthesis, surfactin destroyed cell membrane, and difficidin inhibited the protein biosynthesis of bacteria [23].…”

Section: Discussionmentioning

confidence: 99%

“…These secondary metabolites also induce systemic resistance in plants. Genome sequence analysis indicated that B. velezensis harbors many gene clusters encoding secondary metabolites, i.e., B. velezensis VJH504 contains several gene clusters encoding NRPS, transAT-PKS, T3PKS, and PKS-like types of secondary metabolites [22]; and B. velezensis HNA3 possesses 12 gene clusters related to 14 secondary metabolites with bioactive compounds [23]. Furthermore, B. velezensis shows high hydrolase activity due to the presence of protease, chitinase, cellulase, and glucanase, which is associated with carbon source and cellulose utilization [24].…”

Section: Introductionmentioning

confidence: 99%

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The Bacillus velezensis CYS06 Strain Exhibits Promising Applications in Fighting Grass Carp Bacterial Diseases

Liu,

Jiang,

Ren

et al. 2023

Fishes

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Aeromonas septicemia and columnaris disease are major bacterial diseases in grass carp; however, the drugs currently used to control these diseases pose environmental and health risks. This study aimed to screen for a probiotic Bacillus strain with antagonistic activity to prevent and control bacterial diseases in grass carp and to evaluate the antimicrobial activities, biosafety, and biocontrol effects of this strain. A Bacillus strain with antagonistic activity against Aeromonas hydrophila, obtained from grass carp intestines, was screened, and the isolate CYS06 was identified by analyzing the 16S rRNA and gyrA gene sequences. The antimicrobial spectrum of the strain CYS06 was determined, and the activities of amylase, cellulase, protease, and lipase of the strain CYS06 were determined. The whole genome of the strain CYS06 was sequenced using the nanopore sequencing technology platform, followed by the analysis of the antagonistic substance synthesis gene clusters and CAZy enzyme gene families. The biosafety of the strain CYS06 was evaluated via intraperitoneal injection into healthy grass carp. After the strain CYS06 was fed to the grass carp, its biological control effect on this fish was evaluated through artificial infection experiments. The strain CYS06 was identified as Bacillus velezensis, based on molecular identification, which shows broad antimicrobial activity against various fish pathogens. The strain CYS06 secretes amylase, cellulase, protease, and lipase. The genome size of the strain CYS06 is 3,914,159 bp, and it contains eight antagonistic substance synthesis gene clusters and many CAZy enzymes. The strain CYS06 exhibits high biological safety for grass carp, based on the challenge test. Feeding grass carp with the strain CYS06 for 4 weeks significantly enhanced the resistance of the fish to A. hydrophila. Strain CYS06 could inhibit the growth of Flavobacterium columnare under co-culture and reduce the amount of F. columnare adherence on the gills of grass carp, indicating that CYS06 has good potential for the prevention and control of columnaris disease. In conclusion, we isolated an antagonistic probiotic strain, CYS06, which exhibits a biological control effect on septicemia and columnaris disease caused by Aeromonas spp. and F. columnare in grass carp, respectively. This strain contains many antagonistic substance synthesis-related gene clusters and holds the potential to degrade various types of carbohydrates. As a biological control agent, the strain CYS06 exhibits significant potential for the prevention and control of bacterial diseases in grass carp.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The Bacillus velezensis CYS06 Strain Exhibits Promising Applications in Fighting Grass Carp Bacterial Diseases

Liu,

Jiang,

Ren

et al. 2023

Fishes

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“…Simultaneously, the Fusarium oxysporum f. sp. cucumerinum FJH36 ( Foc FJH36) fungi were isolated and conserved at the Institute of Horticulture, Henan Academy of Agricultural Sciences ( Yang et al, 2023 ).…”

Section: Methodsmentioning

confidence: 99%

“…We measured plant height, stem diameter, leaf area, chlorophyll content, shoot fresh weight, root fresh weight, shoot dry weight and root dry weight on the 20th and 40th days during the whole growth period ( Yang et al, 2023 ; Figure 1D ).…”

Section: Methodsmentioning

confidence: 99%

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Genomic and phenotypic analyses reveal Paenibacillus polymyxa PJH16 is a potential biocontrol agent against cucumber fusarium wilt

Yang,

Jiang,

et al. 2024

Front. Microbiol.

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In recent years, bacterial-based biocontrol agents (BCA) have become a new trend for the control of fungal diseases such as fusarium wilt that seriously threaten the yield and quality of cucumber, which are transmitted through infested soil and water. This study was set out with the aim of figuring the mechanism of the isolated rhizobacterial strain Paenibacillus polymyxa PJH16 in preventing Fusarium oxysporum f. sp. cucumerinum (Foc). Biocontrol and growth-promoting experiments revealed that bacterial strain causes effective inhibition of the fungal disease through a significant growth-promoting ability of plants, and had activities of β-1,3-glucanase, cellulase, amylase and protease. It could produce siderophore and indole-3-acetic acid, too. Using the high-throughput sequencing tool PacBio Sequel II system and the database annotation, the bacterial strain was identified as P. polymyxa PJH16 and contained genes encoding for presence of biofilm formation, antimicrobial peptides, siderophores and hydrolyases. From comparing data between the whole genome of P. polymyxa PJH16 with four closely related P. polymyxa strains, findings revealed markedly the subtle differences in their genome sequences and proposed new antifungal substances present in P. polymyxa PJH16. Therefore, P. polymyxa PJH16 could be utilized in bioengineering a microbial formulation for application as biocontrol agent and bio-stimulant, in the future.

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Characterization and application of Bacillus velezensisD6 co‐producing α‐amylase and protease

Wang,

Bai

et al. 2024

J Sci Food Agric

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BACKGROUNDResearch on the co‐production of multiple enzymes by Bacillus velezensis as a novel species is still a topic that needs to be studied. This study aimed to investigate the fermentation characteristics of B. velezensis D6 co‐producing α‐amylase and protease and to explore their enzymatic properties and applications in fermentation.RESULTSThe maximum co‐production of α‐amylase and protease reached 13.13 ± 0.72 and 2106.63 ± 64.42 U mL−1, respectively, under the optimal fermented conditions (nutrients: 20.0 g L−1 urea, 20.0 g L−1 glucose, 0.7 g L−1 MnCl2; incubation conditions: initial pH 7.0, temperature 41 °C, 8% inoculation size and 30% working volume). Moreover, the genetic co‐expression of α‐amylase and protease increased from 0 to 24 h and then decreased after 36 h at the transcriptional level, which coincided with the growth trend of B. velezensis D6. The optimal reaction temperature of α‐amylase was 55–60 °C, while that of protease was 35–40 °C. The activities of α‐amylase and protease were retained by over 80% after thermal treatment (90 °C, 1 h), which indicated that two enzymes co‐produced by B. velezensis D6 demonstrated excellent thermal stability. Moreover, the two enzymes were stable over a wide pH range (pH 4.0–8.0 for α‐amylase; pH 4.0–9.0 for protease). Finally, the degrees of hydrolysis of corn, rice, sorghum and soybeans by α‐amylase from B. velezensis D6 reached 44.95 ± 2.95%, 57.16 ± 2.75%, 52.53 ± 4.01% and 20.53 ± 2.42%, respectively, suggesting an excellent hydrolysis effect on starchy raw materials. The hydrolysis degrees of mackerel heads and soybeans by protease were 43.93 ± 2.19% and 26.38 ± 1.72%, respectively, which suggested that the protease from B. velezensis D6 preferentially hydrolyzed animal‐based protein.CONCLUSIONThis is a systematic study on the co‐production of α‐amylase and protease by B. velezensis D6, which is crucial in widening the understanding of this species co‐producing multi‐enzymes and in exploring its potential application. © 2024 Society of Chemical Industry.

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Genome sequencing and analysis of Bacillus velezensis VJH504 reveal biocontrol mechanism against cucumber Fusarium wilt

Cited by 8 publications

References 85 publications

The Bacillus velezensis CYS06 Strain Exhibits Promising Applications in Fighting Grass Carp Bacterial Diseases

The Bacillus velezensis CYS06 Strain Exhibits Promising Applications in Fighting Grass Carp Bacterial Diseases

Genomic and phenotypic analyses reveal Paenibacillus polymyxa PJH16 is a potential biocontrol agent against cucumber fusarium wilt

Characterization and application of Bacillus velezensisD6 co‐producing α‐amylase and protease

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