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Isolation, Identification, and Complete Genome Assembly of an Endophytic Bacillus velezensis YB-130, Potential Biocontrol Agent Against Fusarium graminearum.

ABSTRACT: Wheat scab caused by F. graminearum is a highly destructive disease that leads to yield reduction and mycotoxin contamination of grains. In this study, an endophytic bacterium of strain YB-130 was isolated from surface sterilized wheat spikes with scab symptoms and identified as Bacillus velezensis by whole genome annotation, 16S rRNA gene and average nucleotide identities analysis. The whole-genome sequence of strain YB-130 was obtained by PacBio sequencing. 88 putative Carbohydrate-Active Enzymes and 12 gene clusters encoding for secondary metabolites were identified in the YB-130 genome, including one gene cluster for the synthesis of lanthipeptide only found in strain YB-130 genome. In dual cultures, strain YB-130 significantly inhibited the growth of F. graminearum PH-1 and other eight fungal plant pathogens, indicating a broad antifungal activity. Furthermore, strain YB-130 was able to significantly inhibit spore morphology and hyphal development of F. graminearum PH-1. Strain YB-130 also reduced deoxynivalenol production by F. graminearum PH-1 in dual cultures, possibly due to its ability to suppress the expression of tri5, tri3, and tri8 that are required for deoxynivalenol production in F. graminearum. Overall, B. velezensis YB-130 is a promising biological control agent of both F. graminearum infection and mycotoxin production.

SUBMITTER: Xu W

PROVIDER: S-EPMC7744476 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Isolation, Identification, and Complete Genome Assembly of an Endophytic Bacillus velezensis YB-130, Potential Biocontrol Agent Against Fusarium graminearum.

Xu Wen W Zhang Liyong L Goodwin Paul H PH Xia Mingcong M Zhang Jie J Wang Qi Q Liang Juan J Sun Runhong R Wu Chao C Yang Lirong L

Frontiers in microbiology 20201203

Wheat scab caused by F. graminearum is a highly destructive disease that leads to yield reduction and mycotoxin contamination of grains. In this study, an endophytic bacterium of strain YB-130 was isolated from surface sterilized wheat spikes with scab symptoms and identified as Bacillus velezensis by whole genome annotation, 16S rRNA gene and average nucleotide identities analysis. The whole-genome sequence of strain YB-130 was obtained by PacBio sequencing. 88 putative Carbohydra ...[more]

PMID: 33343540

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Project description:The use of biological control agents (BCAs) is a promising alternative control measure for Fusarium crown rot (FCR) of wheat caused by Fusarium pseudograminearum. A bacterial strain, YB-185, was isolated from the soil of wheat plants with FCR and identified as Bacillus velezensis. YB-185 exhibited strong inhibition of F. pseudograminearum mycelial growth and conidial germination in culture. Seed treatment with YB-185 in greenhouse and field resulted in reductions in disease by 66.1% and 57.6%, respectively, along with increased grain yield. Microscopy of infected root tissues confirmed that YB-185 reduced root invasion by F. pseudograminearum. RNA-seq of F. pseudograminearum during co-cultivation with B. velezensis YB-185 revealed 5086 differentially expressed genes (DEGs) compared to the control. Down-regulated DEGs included genes for glucan synthesis, fatty acid synthesis, mechanosensitive ion channels, superoxide dismutase, peroxiredoxin, thioredoxin, and plant-cell-wall-degrading enzymes, whereas up-regulated DEGs included genes for chitin synthesis, ergosterol synthesis, glutathione S-transferase, catalase, and ABC transporters. In addition, fungal cell apoptosis increased significantly, as indicated by TUNEL staining, and the scavenging rate of 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS·+) in the fungus significantly decreased. Thus, F. pseudograminearum may be trying to maintain normal cell functions by increasing cell wall and membrane synthesis, antioxidant and anti-stress responses, detoxification of bacterial antimicrobial compounds, and transportation of damaging compounds from its cells. However, cell death and free radical accumulation still occurred, indicating that the responses were insufficient to prevent cell damage. Bacillus velezensis YB-185 is a promising BCA against FCR that acts by directly damaging F. pseudograminearum, thus reducing its ability to colonize roots and produce symptoms.

Project description:Cucumber Fusarium wilt, caused by Fusarium oxysporum, is a devastating disease of cucumber and leads to enormous economic losses worldwide. The antagonistic bacterium Bacillus velezensis NH-1 suppresses F. oxysporum For a higher biological control effect, control-released microcapsules of NH-1 were prepared using cell immobilization technology. NH-1 cells were embedded in combinations of the biodegradable wall materials sodium alginate, chitosan, and cassava-modified starch to prepare control-released microbiological microcapsules. For the preparation of alginate single-layer microcapsules, the highest embedding rate of 72.60% was obtained by applying 3% sodium alginate and 2% calcium chloride. After the application of monolayer alginate microcapsules in soil, the number of bacterial cells corresponded to a sustained release curve, and the survival rate of NH-1 was higher than the control in which soil was directly irrigated with NH-1 broth. The use of 0.8% chitosan (pH 3.0) and 0.5% cassava-modified starch in the preparation of double-layer and triple-layer microcapsules changed the performance of the microcapsules and increased the embedding rate. After dry storage for 65 days, the number of NH-1 cells was at the highest level in the monolayer microcapsules. In the field experiment, the control efficiency of alginate-coated monolayer microcapsules on Fusarium wilt was 100%, which was significantly higher than for the NH-1 culture and double-layer and triple-layer microcapsules. Collectively, sodium alginate is an ideal wall material for preparing slow-release bacterial microcapsules to control cucumber Fusarium wilt. Monolayer alginate microcapsules retard the release of B. velezensis NH-1 in soils and significantly improve its biocontrol efficiency on cucumber Fusarium wilt.IMPORTANCEBacillus species are often used for the biocontrol of various plant pathogens, but the control efficiency of Bacillus is usually unstable in field experiments. To improve the control efficiency of Bacillus, in this study, microcapsules of Bacillus velezensis strain NH-1 were prepared using different wall materials (sodium alginate, chitosan, and cassava-modified starch). It was found that the control efficiency of alginate-coated monolayer microcapsules on Fusarium wilt was 100% in field experiments, which was higher than for NH-1 culture and double-layer and triple-layer microcapsules. This study provides a new approach for preparing a biocontrol agent against Fusarium wilt with high biocontrol efficiency.

Project description:IntroductionMaize stalk rot (MSR), caused by Fusarium graminearum, is the most serious soil borne disease in maize production, seriously affecting maize yield and quality worldwide. Microbial biocontrol agents are the best means of controlling MSR and reducing the use of chemical fungicides, such as Bacillus spp.Methods and resultsIn the study, a soil-isolated strain B105-8 was identified as B. velezensis (accession No. PP325775.1 and No. PP869695.1), demonstrated a broad spectrum against various pathogens causing maize diseases, which effectively controlled MSR, exhibited a high control efficacy of more than 60% and growth-promoting effect in the pot plant. B105-8 could effectively improve soil urease (S-UE), invertase (S-SC), and catalase (S-CAT) activities. S-NP activity showed an initial increase with a peak of 20,337 nmol/h/g, followed by a decrease, but activity remained significantly better than control treatment with chemical fungicides. The application of B105-8 repaired the damage caused by F. graminearum on soil activity. The antifungal compound B-1, extracted from B105-8, was purified using a protein purifier, revealing inhibitory effects against F. graminearum. Mass spectrometry analysis indicated the potential presence of C14 Bacillomycin, C15 Iturin, C15 Mycosubtilin, C17, and C15 fengycin in B-1. In pot experiments, a 5 μL/mL concentration of B-1 exhibited 69% control on MSR, enhancing maize root elongation, elevation, and fresh weight. At 10 μL/mL, B-1 showed 89.0 and 82.1% inhibition on spore production and mycelial growth, causing hyphal deformities.DiscussionThis study presents the innovative use of B. velezensis, isolated from maize rhizosphere in cold conditions to effectively control MSR caused by F. graminearum. The findings highlight the remarkable regional and adaptive characteristics of this strain, making it an excellent candidate to fight MSR in diverse environments. In conclusion, B. velezensis B105-8 demonstrated potential as a biocontrol agent for MSR.

Dataset Information

Isolation, Identification, and Complete Genome Assembly of an Endophytic Bacillus velezensis YB-130, Potential Biocontrol Agent Against Fusarium graminearum.

Publications

Isolation, Identification, and Complete Genome Assembly of an Endophytic <i>Bacillus velezensis</i> YB-130, Potential Biocontrol Agent Against <i>Fusarium graminearum</i>.

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