Characterization of an antifungal soil bacterium and its antagonistic activities againstFusariumspecies

2003 ◽  
Vol 49 (4) ◽  
pp. 253-262 ◽  
Author(s):  
Yiu-Kwok Chan ◽  
Wayne A McCormick ◽  
Keith A Seifert

Bacteria were isolated from a cultivated soil and screened for antagonistic activity against Fusarium graminearum, a predominant agent of ear rot and head blight in cereal crops. Based on its in vitro effectiveness, isolate D1/2 was selected for characterization and identified as a strain of Bacillus subtilis by phenotypic tests and comparative analysis of its 16S ribosomal RNA gene (rDNA) sequence. It inhibited the mycelial growth of a collection of common fungal phytopathogens, including eight Fusarium species, three other ascomycetes, and one basidiomycete. The cell-free culture filtrate of D1/2 at different dilutions was active against macroconidium germination and hyphal growth of F. graminearum, depending on the initial macroconidium density. It induced the formation of swollen hyphal cells in liquid cultures of this fungus grown from macroconidia. A bioassay also demonstrated that D1/2 offered in planta protection against the damping-off disease in alfalfa seedlings caused by F. graminearum, while the type strain of B. subtilis was ineffective. Hence, B. subtilis D1/2 or its culture filtrate has potential application in controlling plant diseases caused by Fusarium.Key words: antifungal activity, Bacillus subtilis, biological control, biopesticide, Fusarium species.

2020 ◽  
Vol 33 (7) ◽  
pp. 888-901
Author(s):  
Sean P. O’Mara ◽  
Karen Broz ◽  
Marike Boenisch ◽  
Zixuan Zhong ◽  
Yanhong Dong ◽  
...  

The plant-pathogenic fungus Fusarium graminearum, causal agent of Fusarium head blight (FHB) disease on small grain cereals, produces toxic trichothecenes that require facilitated export for full virulence. Two potential modes of mycotoxin transport are membrane-bound transporters, which move toxins across cellular membranes, and N-ethylmaleimide-sensitive factor attachment receptor (SNARE)-mediated vesicular transport, by which toxins may be packaged as cargo in vesicles bound for organelles or the plasma membrane. In this study, we show that deletion of a gene (Sso2) for a subapically localized t-SNARE protein results in growth alteration, increased sensitivity to xenobiotics, altered gene expression profiles, and reduced deoxynivalenol (DON) accumulation in vitro and in planta as well as reduced FHB symptoms on wheat. A double deletion mutant generated by crossing the ∆sso2 deletion mutant with an ATP-binding cassette transporter deletion mutant (∆abc1) resulted in an additive reduction in DON accumulation and almost complete loss of FHB symptoms in planta. These results suggest an important role of Sso2-mediated subapical exocytosis in FHB progression and xenobiotic defense and are the first report of an additive reduction in F. graminearum DON accumulation upon deletion of two distinct modes of cellular export. This research provides useful information which may aid in formulating novel management plans of FHB or other destructive plant diseases.


2021 ◽  
Author(s):  
Mohamed Hafez ◽  
Ryan Gourlie ◽  
Melissa Telfer ◽  
Nicola Schatz ◽  
Kelly Turkington ◽  
...  

Fusarium head blight (FHB) and Fusarium crown and root rot (FCRR) are major wheat diseases worldwide. In this study, wheat node and grain samples were collected from four representative sites across the western Canadian prairies in 2018 growing season to characterize the major Fusarium spp. and other mycobiota associated with wheat in these regions. In total, 994 fungal isolates were recovered and based on culture and molecular diagnostic methods, three genera constituted over 90% of all fungal isolates, these were Alternaria (39.6%), Fusarium (27.8%), and Parastagonospora (23.9%). A qPCR was developed to quantify the most frequently isolated Fusarium spp. in infected wheat tissues: F. avenaceum, F. culmorum, F. graminearum and F. poae. The qPCR specificity was validated in silico, in vitro and in planta and proved specific to the target species.. The qPCR results showed that F. graminearum was not detected frequently in the four tested locations in this study. F. poae was the most abundant Fusarium species in grain samples in all tested locations. However, in node samples, F. culmorum (Beaverlodge and Scott) and F. avenaceum (Lacombe and Lethbridge) were the most abundant species. Trichothecene genotyping, showed that the 3ADON is the most dominant trichothecene genotype (68%), followed by type-A trichothecenes (29.5%), while the 15ADON trichothecene genotype was least dominant (2.5%) and the NIV genotype was not detected. Moreover, a total of 129 TEF1α sequences from nine Fusarium spp. were compared at haplotype level to evaluate genetic variability and haplotype distribution. F. avenaceum and F.poae exhibited higher diversity.


2021 ◽  
Vol 12 ◽  
Author(s):  
Catarina Leal ◽  
Nicolas Richet ◽  
Jean-François Guise ◽  
David Gramaje ◽  
Josep Armengol ◽  
...  

Grapevine trunk diseases (GTDs) are a big threat for global viticulture. Without effective chemicals, biocontrol strategies are developed as alternatives to better cope with environmental concerns. A combination of biological control agents (BCAs) could even improve sustainable disease management through complementary ways of protection. In this study, we evaluated the combination of Bacillus subtilis (Bs) PTA-271 and Trichoderma atroviride (Ta) SC1 for the protection of Chardonnay and Tempranillo rootlings against Neofusicoccum parvum Bt67, an aggressive pathogen associated to Botryosphaeria dieback (BD). Indirect benefits offered by each BCA and their combination were then characterized in planta, as well as their direct benefits in vitro. Results provide evidence that (1) the cultivar contributes to the beneficial effects of Bs PTA-271 and Ta SC1 against N. parvum, and that (2) the in vitro BCA mutual antagonism switches to the strongest fungistatic effect toward Np-Bt67 in a three-way confrontation test. We also report for the first time the beneficial potential of a combination of BCA against Np-Bt67 especially in Tempranillo. Our findings highlight a common feature for both cultivars: salicylic acid (SA)-dependent defenses were strongly decreased in plants protected by the BCA, in contrast with symptomatic ones. We thus suggest that (1) the high basal expression of SA-dependent defenses in Tempranillo explains its highest susceptibility to N. parvum, and that (2) the cultivar-specific responses to the beneficial Bs PTA-271 and Ta SC1 remain to be further investigated.


Author(s):  
Tony Twamley ◽  
Mark Gaffney ◽  
Angela Feechan

AbstractFusarium graminearum and Zymoseptoria tritici cause economically important diseases of wheat. F. graminearum is one of the primary causal agents of Fusarium head blight (FHB) and Z. tritici is the causal agent of Septoria tritici blotch (STB). Alternative control methods are required in the face of fungicide resistance and EU legislation which seek to cut pesticide use by 2030. Both fungal pathogens have been described as either hemibiotrophs or necrotrophs. A microbial fermentation-based product (MFP) was previously demonstrated to control the biotrophic pathogen powdery mildew, on wheat. Here we investigated if MFP would be effective against the non-biotrophic fungal pathogens of wheat, F. graminearum and Z. tritici. We assessed the impact of MFP on fungal growth, disease control and also evaluated the individual constituent parts of MFP. Antifungal activity towards both pathogens was found in vitro but MFP only significantly decreased disease symptoms of FHB in planta. In addition, MFP was found to improve the grain number and weight, of uninfected and F. graminearum infected wheat heads.


2022 ◽  
Vol 8 (1) ◽  
pp. 80
Author(s):  
Yongmei Li ◽  
Mengyuan Xia ◽  
Pengbo He ◽  
Qiaoming Yang ◽  
Yixin Wu ◽  
...  

Citrus is among the most important plants in the fruit industry severely infected with pathogens. Citrus green mold caused by Penicillium digitatum is one of the most devastating diseases during post-harvest stages of citrus fruit. In this study, a potential endophyte Bacillus subtilis L1-21, isolated from healthy citrus plants, was assessed for its biocontrol activity against the pathogen P. digitatum. Based on an in vitro crosstalk assay, we suggested that B. subtilis L1-21 inhibits the pathogen with an inhibition zone of 3.51 ± 0.08 cm. Biocontrol efficacy was highest for the fermented culture filtrate of B. subtilis L1-21. Additionally, using GC-MS analysis, 13 compounds were detected in the extract of this endophyte. The culture filtrate in Landy medium could enlarge and deform pathogen spores and prevent them from developing into normal mycelium. Accordingly, the Landy culture filtrate of B. subtilis L1-21 was stable in the temperature range of 4–90 °C and pH of 3–11. Further, MALDI-TOF-MS for B. subtilis L1-21 detected surfactin, fengycin, bacillaene and bacilysin as potential antifungal compounds. GFP-tagged B. subtilis L1-21 easily colonized in citrus fruit peel and pulp, suggesting its role in eliminating the fungal pathogen. Altogether, it is highly expected that the production of antifungal compounds, and the colonization potential of B. subtilis L1-21 are required against the post-harvest P. digitatum pathogen on citrus fruit.


Plant Disease ◽  
2009 ◽  
Vol 93 (10) ◽  
pp. 1037-1043 ◽  
Author(s):  
Young-Ki Jo ◽  
Byung H. Kim ◽  
Geunhwa Jung

Silver in ionic or nanoparticle forms has a high antimicrobial activity and is therefore widely used for various sterilization purposes including materials of medical devices and water sanitization. There have been relatively few studies on the applicability of silver to control plant diseases. Various forms of silver ions and nanoparticles were tested in the current study to examine the antifungal activity on two plant-pathogenic fungi, Bipolaris sorokiniana and Magnaporthe grisea. In vitro petri dish assays indicated that silver ions and nanoparticles had a significant effect on the colony formation of these two pathogens. Effective concentrations of the silver compounds inhibiting colony formation by 50% (EC50) were higher for B. sorokiniana than for M. grisea. The inhibitory effect on colony formation significantly diminished after silver cations were neutralized with chloride ions. Growth chamber inoculation assays further confirmed that both ionic and nanoparticle silver significantly reduced these two fungal diseases on perennial ryegrass (Lolium perenne). Particularly, silver ions and nanoparticles effectively reduced disease severity with an application at 3 h before spore inoculation, but their efficacy significantly diminished when applied at 24 h after inoculation. The in vitro and in planta evaluations of silver indicated that both silver ions and nanoparticles influence colony formation of spores and disease progress of plant-pathogenic fungi. In planta efficacy of silver ions and nanoparticles is much greater with preventative application, which may promote the direct contact of silver with spores and germ tubes, and inhibit their viability.


Plant Disease ◽  
2021 ◽  
Vol 105 (1) ◽  
pp. 134-143
Author(s):  
Namgyu Kim ◽  
Mohamed Mannaa ◽  
Juyun Kim ◽  
Ji-Eun Ra ◽  
Sang-Min Kim ◽  
...  

Burkholderia glumae, B. plantarii, and B. gladioli are responsible for serious diseases in rice crops and co-occurrence among them has been reported. In this study, in vitro assays revealed antagonistic activity among these organisms, with B. gladioli demonstrating strong inhibition of B. glumae and B. plantarii. Strains of B. glumae and B. plantarii that express green fluorescent protein were constructed and used for cocultivation assays with B. gladioli, which confirmed the strong inhibitory activity of B. gladioli. Cell-free supernatants from each species were tested against cultures of counterpart species to evaluate the potential to inhibit bacterial growth. To investigate the inhibitory activity of B. gladioli on B. glumae and B. plantarii in rice, rice plant assays were performed and quantitative PCR (qPCR) assays were developed for in planta bacterial quantification. The results indicated that coinoculation with B. gladioli leads to significantly reduced disease severity and colonization of rice tissues compared with single inoculation with B. glumae or B. plantarii. This study demonstrates the interactions among three rice-pathogenic Burkholderia species and strong antagonistic activity of B. gladioli in vitro and in planta. The qPCR assays developed here could be applied for accurate quantification of these organisms from in planta samples in future studies.


2002 ◽  
Vol 2002 ◽  
pp. 80-80
Author(s):  
K. Hillman

The ability of the porcine intestinal microflora to resist the establishment of pathogenic bacteria has been demonstrated previously (Hillman et al, 1994). Subsequent work has shown that certain intestinal lactobacilli react to the presence of a culture filtrate derived from a coliform pathogen by increasing their antagonistic activity towards that pathogen (Hillman and Robertson, unpublished), indicating the presence of a quorum-sensing or related mechanism. The current experiment was devised to determine whether a similar effect could be produced within the entire porcine colonic microflora, using an in vitro simulation system.


2021 ◽  
Vol 48 (3) ◽  
Author(s):  
Hind Lahmyed ◽  
◽  
Rachid Bouharroud ◽  
Redouan Qessaoui ◽  
Abdelhadi Ajerrar ◽  
...  

The present work aims to isolate actinomycete bacteria with antagonistic abilities towards Botrytis cinerea, the causal agent of gray mold, from a soil sample collected from the rhizosphere of a healthy tomato grove. In vitro confrontation led to the isolation of 104 actinomycete isolates; fifteen isolates have shown the most significant mortality rate of the mycelial growth of B. cinerea (>50%). Based on the results of this screening, representative strains were selected to verify their in vivo antagonistic activity on tomato fruits; the reduction of B. cinerea has a percentage ranging from 52.38% to 96.19%. Furthermore, the actinomycete isolates were evaluated for their plant growth-promoting (PGP) properties and their ability to produce biocontrol-related extracellular enzymes viz., amylase, protease, cellulase, chitinase, esterases, and lecithinase. Indeed, Ac70 showed high β-1,3-glucanase activity and siderophore production (17U/ml and 43% respectively), and the highest chitinase activity (39μmol/ml) was observed for Ac24. These results indicated that these actinomycetes might potentially control gray mold caused by B. cinerea on tomato fruits. Investigations on enhancing the efficacy and survival of the biocontrol agent in planta and finding out the best formulation are recommended for future research.


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