Antagonistic Bacteria Research Articles

Biocontrol Capabilities of Bacillus subtilis E11 against Aspergillus flavus In Vitro and for Dried Red Chili (Capsicum annuum L.).

As a condiment with extensive nutritional value, chili is easy to be contaminated by Aspergillus flavus (A. flavus) during field, transportation, and storage. This study aimed to solve the contamination of dried red chili caused by A. flavus by inhibiting the growth of A. flavus and detoxifying aflatoxin B1 (AFB1). In this study, Bacillus subtilis E11 (B. subtilis) screened from 63 candidate antagonistic bacteria exhibited the strongest antifungal ability, which could not only inhibit 64.27% of A. flavus but could also remove 81.34% of AFB1 at 24 h. Notably, scanning electron microscopy (SEM) showed that B. subtilis E11 cells could resist a higher concentration of AFB1, and the fermentation supernatant of B. subtilis E11 could deform the mycelia of A. flavus. After 10 days of coculture with B. subtilis E11 on dried red chili inoculated with A. flavus, the mycelia of A. flavus were almost completely inhibited, and the yield of AFB1 was significantly reduced. Our study first concentrated on the use of B. subtilis as a biocontrol agent for dried red chili, which could not only enrich the resources of microbial strains for controlling A. flavus but also could provide theoretical guidance to prolong the shelf life of dried red chili.

Screening of potential bacterial isolates against Phytophthora capsici and its plant growth-promoting effect on pepper plants

Phytophthora blight, is the most devastating disease of pepper production. To increase agricultural yield and control diseases, many countries in the world use synthetic agrochemicals. However, it has negative effects on human health and the environment, but there are few reports on its biological control. Out of sixty bacterial strains, four that showed biocontrol and plant growth promotion, namely Enterobacter hormaechei, Rhizobium sp., Pseudomonas fluorescens, and undetermined (AAULE51), which were isolated and identified using biochemical tests and 16S RNA sequences. All bacterial strains that showed greater than 55.5% growth inhibition of Phytophthora capsici in dual culture were selected for greenhouse and field studies. Also, the antimicrobial activity of the cell-free filtrate of the strains were assessed. AAULE51 was found to be the best inhibitor against P. capsici, ranging from 90.34 to 100% at a concentration of 20%–75%. The effects of bacterial strains on seed germination, vigor index, seedling height, and fresh weight of pepper plants were measured in vitro and showed 70−90.6%, 1069.6−2650%, 37−48 mm, and 64−70 mg, respectively. In greenhouse and field studies the biocontrol efficacy of antagonistic bacteria against P.capsici ranged from 0.6% to 94.2%, and 11%–84.6% respectively. The maximum reduction in disease severity was shown when treated with AAULE41+ AAULE51 (61.1%–94.2%). Therefore, these results suggest that selected potential bacterial strains are good candidates for agricultural biofertilizers and biopesticides.

Antagonistic bacteria against Fusarium spp. isolated from sclerotia of Claviceps gigantea in maize (Zea mays)

Fungal sclerotia house native bacterial populations of interest for biological control of plant pathogenic fungi. The objectives of this research were: i) to explore the bacterial populations associated with Claviceps gigantea sclerotia in maize from six locations in the State of Mexico, ii) to evaluate the in vitro antagonism of these bacterial populations against three species of Fusarium phytopathogens of maize, iii) to molecularly identify the more efficient antagonists and characterize the in vitro production of metabolites. Bacterial populations in the sclerotia were calculated by direct plate count; in vitro antagonism against Fusarium graminearum, F. sublgutinans, and F. verticillioides was evaluated by confrontation in Waksman agar medium. Antagonistic bacteria were identified by partial sequencing of 16S rRNA gene and evaluated in vitro for the production of indole-3 acetic acid, siderophores, lipolytic and proteolytic activity, and mineral phosphate solubilization. The bacterial density ranged from 2.023 to 2.397 Log10UFC g-1 of sclerotia. Twenty-two strains showed in vitro antagonism against at least one Fusarium species and were identified as members of the genera Bacillus, Delftia, Micromonospora, Pseudomonas, Sphingobacterium, Staphylococcus, and Stenotrophomonas. The 22 antagonists showed in vitro lipolytic, proteolytic, siderophore-producing and phosphate-solubilizing activity; only 12 (55%) produced indole-3-acetic acid. Bacillus subtilis (BA1), Pseudomonas syringae (BA2), and Bacillus amyloliquefaciens (BA18) strains were antagonists against the three Fusarium species and produced all the metabolites evaluated.

Foreign research experience of the effect of probiotic agent on the intestinal microbiome of poultry

The intensive development of industrial animal husbandry and poultry farming, along with the ban on the use of antibiotics for non-medicinal purposes, requires a revision of stimulating and therapeutic and preventive approaches aimed at increasing productivity, protecting against infections and improving other production parameters, such as feed digestibility and improving the efficiency of digestion, stimulating growth and development, the quality of meat, milk, eggs. In this regard, probiotic, prebiotic and synbiotic drugs and feed additives have become widespread. Funds based on antagonistic bacteria and representatives of symbiotic microflora are actively being introduced into poultry technology, a wide domestic research experience reveals their prospects. Used mainly to maintain the balance of the intestinal microbiota, they turn out to be an effective method of combating opportunistic pathogens.At the same time, in the countries of developed industrial poultry farming, products based on probiotic cultures have not been widely used, and a number of sources are skeptical about the ability of probiotics to influence the intestinal microbiome of poultry. This article discusses various modern studies that carry the opinions of foreign authors about the effectiveness of probiotics and their effect on the intestinal microbiome of poultry.

Biocontrol Stem-end Rot Disease Agent, Alternaria alternata YZU, on Pitaya by Soil Phosphate Solubilizing Bacteria

Biocontrol of stem end disease agent, Alternaria alternata YZU, on pitaya is more interested in altering the application of chemical pesticides. This study was conducted to characterize antagonistic phosphate solubilizing bacteria (PSB) from rhizosphere soil for their biocontrol activities against A. alternata YZU under laboratory and greenhouse conditions. Six PSB isolated from 31 rhizosphere soil samples were tested for inhibiting the mycelial growth of A. alternata YZU in dual cultures. Six isolates were determined to inhibit the mycelial growth of A. alternata YZU. Among them, PSB31 presented the highest level of antagonistic activity against A. alternata YZU with a mean inhibition diameter of 0.64 ± 0.02 cm, while the other strains, including PSB11, PSB21, PSB41, PSB51, and PSB61 presented a weaker inhibition. The results also showed that the strain PSB31 was identified as Bacillus sp. strain PSB31 (Accession number: ON422095) and could control the mycelial growth of A. alternata YZU by secreting antifungal metabolites. Moreover, an in vivo antagonistic experiment of PSB31 on pitaya twigs showed a significant reduction of lesions on twigs than the control. The results suggested that the isolated PSB31 is a potential biological control agent. Further studies should be done to identify the biochemical basis of their activity against A. alternata YZU.

Biocontrol potential and action mechanism of Bacillus amyloliquefaciens DB2 on Bipolaris sorokiniana.

Bipolaris sorokiniana is the popular pathogenic fungi fungus which lead to common root rot and leaf spot on wheat. Generally, chemical fungicides are used to control diseases. However, the environmental pollution resulting from fungicides should not be ignored. It is important to study the mode of antagonistic action between biocontrol microbes and plant pathogens to design efficient biocontrol strategies. An antagonistic bacterium DB2 was isolated and identified as Bacillus amyloliquefaciens. The inhibition rate of cell-free culture filtrate (CF, 20%, v/v) of DB2 against B. sorokiniana reached 92.67%. Light microscopy and scanning electron microscopy (SEM) showed that the CF significantly altered the mycelial morphology of B. sorokiniana and disrupted cellular integrity. Fluorescence microscopy showed that culture filtrate destroyed mycelial cell membrane integrity, decreased the mitochondrial transmembrane potential, induced reactive oxygen species (ROS) accumulation, and nuclear damage which caused cell death in B. sorokiniana. Moreover, the strain exhibited considerable production of protease and amylase, and showed a significant siderophore and indole-3-acetic acid (IAA) production. In the detached leaves and potted plants control assay, B. amyloliquefacien DB2 had remarkable inhibition activity against B. sorokiniana and the pot control efficacy was 75.22%. Furthermore, DB2 suspension had a significant promotion for wheat seedlings growth. B. amyloliquefaciens DB2 can be taken as a potential biocontrol agent to inhibit B. sorokiniana on wheat and promote wheat growth.

Links between Soil Bacteriobiomes and Fungistasis toward Fungi Infecting the Colorado Potato Beetle.

Entomopathogenic fungi can be inhibited by different soil microorganisms, but the effect of a soil microbiota on fungal growth, survival, and infectivity toward insects is insufficiently understood. We investigated the level of fungistasis toward Metarhizium robertsii and Beauveria bassiana in soils of conventional potato fields and kitchen potato gardens. Agar diffusion methods, 16S rDNA metabarcoding, bacterial DNA quantification, and assays of Leptinotarsa decemlineata survival in soils inoculated with fungal conidia were used. Soils of kitchen gardens showed stronger fungistasis toward M. robertsii and B. bassiana and at the same time the highest density of the fungi compared to soils of conventional fields. The fungistasis level depended on the quantity of bacterial DNA and relative abundance of Bacillus, Streptomyces, and some Proteobacteria, whose abundance levels were the highest in kitchen garden soils. Cultivable isolates of bacilli exhibited antagonism to both fungi in vitro. Assays involving inoculation of nonsterile soils with B. bassiana conidia showed trends toward elevated mortality of L. decemlineata in highly fungistatic soils compared to low-fungistasis ones. Introduction of antagonistic bacilli into sterile soil did not significantly change infectivity of B. bassiana toward the insect. The results support the idea that entomopathogenic fungi can infect insects within a hypogean habitat despite high abundance and diversity of soil antagonistic bacteria.

History of Discovery and Environmental Role of Ice Nucleating Bacteria.

The phenomenon of biological ice nucleation that is exhibited by a variety of bacteria is a fascinating phenotype, which has been shown to incite frost damage to frost-sensitive plants and has been proposed to contribute to atmospheric processes that affect the water cycle and earth's radiation balance. This review explores the several possible drivers for the evolutionary origin of the ice nucleation phenotype. These bacteria and the gene required for this phenotype have also been exploited in processes as diverse as reporter gene assays to assess environmentally responsive gene expression in various plant pathogenic and environmental bacteria and in the detection of foodborne human pathogens when coupled with host-specific bacteriophage, whereas ice nucleating bacteria themselves have been exploited in the production of artificial snow for recreation and oil exploration and in the process of freezing of various food products. This review also examines the historical development of our understanding of ice nucleating bacteria, details of the genetic determinants of ice nucleation, and features of the aggregates of membrane-bound ice nucleation protein necessary for catalyzing ice. Lastly, this review also explores the role of these bacteria in limiting the supercooling ability of plants and the strategies and limitations of avoiding plant frost damage by managing these bacterial populations by bactericides, antagonistic bacteria, or cultural control strategies.

Identification and characterization of biocontrol agent Lysinibacillus boronitolerans P42 against Cerrena unicolor that causes root rot of arecanut palm

The arecanut palm is one of the most important industrial crops in tropical area around the world. The root rot of arecanut palm, which is caused by Cerrena unicolor, has led to heavy economic losses and restricted greatly the development of arecanut industry, especially in Hainan province of China. The common use of chemical agents has worsened the problems of the emergence of resistant pathogens and the pollution of agricultural environment. This study aims to screen and identify a more effective and environment friendly biocontrol method for the prevention and treatment of root rot of arecanut palm. The mycelium growth rate is investigated to select antagonistic bacteria from tropical crop rotation fields which show improved resistance against soil-borne pathogens, and the strain P42 is revealed with the strongest antagonistic effects (82.18%). Based on 16s rDNA sequence analysis, the strain P42 is identified as Lysinibacillus boronitolerans. In vitro antimicrobial activity shows that the strain P42 exhibits broad-spectrum antagonistic activity against a wide variety of tropical agricultural fungal pathogens, including Cerrena unicolor, Magnaporthe oryzea, Botryodiplodia theobromae, Neoscytalidium dimidiatum, Thanatephorus cucumeris, Fusarium oxysporum, and Botrytis cinereaPer.. The antagonistic activity of the culture of P42 is tolerant to common proteases, longer storage time, and temperature range of 40-121°C; and is significantly influenced by alkaline (7-9) and acidic (1-2) pH, as well as by ultraviolet ray treatment for more than 30min. The investigation on the antagonistic activity of the crude extract of fermentation filtrate indicates that the active compounds might be lipopeptides, polyketones, or proteins. To our knowledge, this is the first report of L. boronitolerans as potential bio-reagents for controlling root rot of arecanut palm caused by Cerrena unicolor.

Бактерии р. Bacillus – антагонисты патогенов аквакультуры

Due to the increasing resistance of aquaculture pathogens to antibiotics, the search for new approaches to the control of infectious diseases is of particular importance. A promising approach is the development of probiotic preparations based on antagonist bacteria. The paper studied the antagonistic activity of Bacillus strains that are promising for developing probiotics for fish. The strains were isolated from the bottom sediments of the Don River in the area of the Donskoy Fish Reserve, as well as from the intestines of cyprinids of natural populations, namely, from silver carp (Cyprinus gibelio), roach (Rutilus heckelii), and bream (Abramis brama). The inhibitory effect of Bacillus strains against causative agents of aeromonosis, pseudomonosis and bacterial hemorrhagic septicemia was studied by the method of delayed antagonism. Aeromonas and Pseudomonas species pathogenic for fish were isolated from sturgeons, cyprinids and salmons grown in pond farms and recirculating aquaculture systems in the Krasnodar Region, Rostov Region and the Republic of Adygea. It has been shown that sensitivity to the action of bacilli is not only a species, but also a strain-specific trait among bacterial pathogens of fish. Five strains were identified that exhibited the maximum antagonistic activity against the studied pathogens, and their species identification was carried out by molecular biological and mass spectrometric methods. It has been established that the most pronounced antagonism is exhibited by strains of bacilli isolated from bottom sediments. The growth of all studied strains of fish pathogenic species of Pseudomonas was suppressed only the B. subtilis R4 strain. Thus, strains of aerobic spore-forming bacteria suitable for the creation of probiotic preparations for aquaculture objects have been isolated and characterized.

Evaluation of Antifungal Activity from Bacillus subtilis strain AS80 Against Pathogenic Fungi Isolated from Hass Avocado Fruits

F ruit rot and anthracnose are the main diseases in avocado fruits caused by fungal species. Solving this problem with chemical reagents has led to human health and pollution problems. Therefore, this study was aimed to select antagonistic bacteria to control those pathogenic fungi. The pathogenic fungi were isolated from avocado fruit occurred fruit rot or anthracnose symptoms. Based on morphological characteristics and the BLASTn results of sequences of the internal transcribed spacer of ribosomal DNA, the obtained fungal isolates belonged to three fungal genera, namely Fusarium, Lasiodiplodia and Colletotrichum, which was the most commonly occurring. Pathogenicity tests were evaluated on avocado fruit. Then the rhizosphere bacteria were isolated from the avocado rhizosphere. To study antagonistic activity, 101 bacterial isolates were cultured with five fungal pathogens using dual culture technique. The bacterial isolate AS80 exhibited high potential to against Fusarium sp. (61.7%), Lasiodiplodia sp. (66.7%) and Colletotrichum sp. (74.1%). The strain AS80 was used to evaluate the efficacy control of Fusarium sp. on “Hass” avocado fruit. The in vivo evaluation revealed that AS80 showed high efficacy against Fusarium sp. The 16s ribosomal DNA sequencing indicated that AS80 was similar to Bacillus subtilis (99.53 %). Further study under field conditions should be done to validate their efficacy.

Mitigation of antibiotic resistome in swine manure by black soldier fly larval conversion combined with composting

The increasing prevalence of antibiotic resistance genes (ARGs) in animal manure has attracted considerable attention because of their potential contribution to the development of multidrug resistance worldwide. Insect technology may be a promising alternative for the rapid attenuation of ARGs in manure; however, the underlying mechanism remains unclear. This study aimed to evaluate the effects of black soldier fly (BSF, Hermetia illucens [L.]) larvae conversion combined with composting on ARGs dynamics in swine manure and to uncover the mechanisms through metagenomic analysis. Compared to natural composting (i.e. without BSF), BSFL conversion combined with composting reduced the absolute abundance of ARGs by 93.2 % within 28 days. The rapid degradation of antibiotics and nutrient reformulation during BSFL conversion combined with composting indirectly altered manure bacterial communities, resulting in a lower abundance and richness of ARGs. The number of main antibiotic-resistant bacteria (e.g., Prevotella, Ruminococcus) decreased by 74.9 %, while their potential antagonistic bacteria (e.g., Bacillus, Pseudomonas) increased by 128.7 %. The number of antibiotic-resistant pathogenic bacteria (e.g., Selenomonas, Paenalcaligenes) decreased by 88.3 %, and the average number of ARGs carried by each human pathogenic bacterial genus declined by 55.8 %. BSF larvae gut microbiota (e.g., Clostridium butyricum, C. bornimense) could help reduce the risk of multidrug-resistant pathogens. These results provide insight into a novel approach to mitigate multidrug resistance from the animal industry in the environment by using insect technology combined with composting, in particular in light of the global “One Health” requirements.

Anaerobic Soil Disinfestation Promotes Soil Microbial Stability and Antagonistic Bacteria Abundance in Greenhouse Vegetable Production Systems

Excessive water and fertiliser inputs, as well as long-term monocropping, not only cause resource waste and environmental pollution but also drive soil degradation and the occurrence of soil-borne diseases. Anaerobic soil disinfestation (ASD) is a widely used technique in greenhouse vegetable production to replace the use of agrochemicals in disinfestation and improve product quality. While the short-term effects of ASD on the soil microbial community have been well documented in the past 15 years, only a few studies have investigated the multiseason effects of ASD, particularly on the soil microbial community composition and stability, as well as on pathogens and antagonistic microorganisms. Field experiments were conducted in three adjacent greenhouses used for tomato production for at least 20 years. Three treatments were included: CK (control: no irrigation, no plastic film covering, incorporation of chicken manure), ASD (irrigation, plastic film covering, and incorporation of rice husks), and ASD+M (ASD plus incorporation of chicken manure). Results showed that (1) ASD significantly reduced the diversity of soil bacteria and fungi and improved the complexity and stability of the soil bacterial community at the end of the ASD, but the soil microbial diversity recovered to the level before the experiment after 1.5 years. (2) Compared with CK, ASD significantly increased the relative abundance of antagonistic bacteria Bacillus, Paenibacillus and Streptomyces, decreased the relative abundance of pathogens Fusarium and the quantity of nematodes and could still effectively eliminate soil pathogens after 1.5 years. (3) Chicken manure application did not increase the pathogenic microorganisms Fusarium and nematodes, but it significantly decreased the relative abundance of antagonistic bacteria. Our results highlight that ASD not only showed an inhibitory effect on soil-borne diseases after 1.5 years but also significantly increased the relative abundance of antagonistic bacteria. However, the additional incorporation of chicken manure for ASD should be avoided due to its negative effects on the abundance of antagonistic bacteria and its contribution to environmental pollution due to N leaching and increased emissions of GHG N2O.

Combating the Dust Devil: Utilizing Naturally Occurring Soil Microbes in Arizona to Inhibit the Growth of Coccidioides spp., the Causative Agent of Valley Fever.

The fungal disease Valley fever causes a significant medical and financial burden for affected people in the endemic region, and this burden is on the rise. Despite the medical importance of this disease, little is known about ecological factors that influence the geographic point sources of high abundance of the pathogens Coccidioides posadasii and C. immitis, such as competition with co-occurring soil microbes. These "hot spots", for instance, those in southern Arizona, are areas in which humans are at greater risk of being infected with the fungus due to consistent exposure. The aim of this study was to isolate native microbes from soils collected from Tucson, Arizona (endemic area for C. posadasii) and characterize their relationship (antagonistic, synergistic, or neutral) to the fungal pathogen with in vitro challenge assays. Secreted metabolites from the microbes were extracted and described using analytical techniques including high-performance liquid chromatography (HPLC) and mass spectrometry. Bacteria belonging to the genus Bacillus and fungi in the Fennellomyces and Ovatospora genera were shown to significantly decrease the growth of Coccidioides spp. In vitro. In contrast, other bacteria in the Brevibacillus genus, as well as one species of Bacillus bacteria, were shown to promote growth of Coccidioides when directly challenged. The metabolites secreted from the antagonistic bacteria were described using HPLC and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The microbes identified in this study as antagonists to Coccidioides and/or the metabolites they secrete have the potential to be used as natural biocontrol agents to limit the amount of fungal burden at geographic point sources, and therefore limit the potential for human infection.

Biostimulant Activity of Silicate Compounds and Antagonistic Bacteria on Physiological Growth Enhancement and Resistance of Banana to Fusarium Wilt Disease.

Biostimulants such as silicate (SiO32-) compounds and antagonistic bacteria can alter soil microbial communities and enhance plant resistance to the pathogens and Fusarium oxysporum f. sp. cubense (FOC), the causal agent of Fusarium wilt disease in bananas. A study was conducted to investigate the biostimulating effects of SiO32- compounds and antagonistic bacteria on plant growth and resistance of the banana to Fusarium wilt disease. Two separate experiments with a similar experimental setup were conducted at the University of Putra Malaysia (UPM), Selangor. Both experiments were arranged in a split-plot randomized complete block design (RCBD) with four replicates. SiO32- compounds were prepared at a constant concentration of 1%. Potassium silicate (K2SiO3) was applied on soil uninoculated with FOC, and sodium silicate (Na2SiO3) was applied to FOC-contaminated soil before integrating with antagonistic bacteria; without Bacillus spp. ((0B)-control), Bacillus subtilis (BS), and Bacillus thuringiensis (BT). Four levels of application volume of SiO32- compounds [0, 20, 40, 60 mL) were used. Results showed that the integration of SiO32- compounds with BS (108 CFU mL-1) enhanced the physiological growth performance of bananas. Soil application of 28.86 mL of K2SiO3 with BS enhanced the height of the pseudo-stem by 27.91 cm. Application of Na2SiO3 and BS significantly reduced the Fusarium wilt incidence in bananas by 56.25%. However, it was recommended that infected roots of bananas should be treated with 17.36 mL of Na2SiO3 with BS to stimulate better growth performance.

Cooperative Action of Fulvic Acid and Bacillus paralicheniformis Ferment in Regulating Soil Microbiota and Improving Soil Fertility and Plant Resistance to Bacterial Wilt Disease.

Excessive continuous cropping and soil degradation, such as acidification, hardening, fertility decline, and the degradation of microbial community, lead to the epidemic of soilborne diseases and cause great loss in agriculture production. Application of fulvic acid can improve the growth and yield of various crops and effectively suppress soilborne plant diseases. Bacillus paralicheniformis strain 285-3 producing poly-gamma-glutamic acid is used to remove the organic acid that can cause soil acidification and increase the fertilizer effect of fulvic acid and the effect of improving soil quality and inhibiting soilborne disease. In field experiments, the application of fulvic acid and Bacillus paralicheniformis ferment effectively reduced the incidence of bacterial wilt disease and improved soil fertility. Both fulvic acid powder and B. paralicheniformis ferment improved soil microbial diversity and increased the complexity and stability of the microbial network. For B. paralicheniformis ferment, the molecular weight of poly-gamma-glutamic acid became smaller after heating, which could better improve the soil microbial community and network structure. In fulvic acid and B. paralicheniformis ferment-treated soils, the synergistic interaction between microorganisms increased and the number of keystone microorganisms increased, which included antagonistic bacteria and plant growth-promoting bacteria. Changes in the microbial community and network structure were the main reason for the reduced incidence of bacterial wilt disease. Application of fulvic acid and Bacillus paralicheniformis ferment improved soil physicochemical properties and effectively controlled bacterial wilt disease by changing microbial community and network structure and enriching antagonistic and beneficial bacteria. IMPORTANCE Continuous cropping tobacco has led to soil degradation and caused soilborne bacterial wilt disease. Fulvic acid as a biostimulator was applied to restore soil and control bacterial wilt disease. For improving its effect, fulvic acid was fermented with Bacillus paralicheniformis strain 285-3 producing poly-gamma-glutamic acid. Fulvic acid and B. paralicheniformis ferment inhibited bacterial wilt disease, improved soil quality, enriched beneficial bacteria, and increased microbial diversity and microbial network complexity. Some keystone microorganisms in fulvic acid and B. paralicheniformis ferment-treated soils had potential antimicrobial activity and plant growth-promoting attributes. Fulvic acid and B. paralicheniformis 285-3 ferment could be used to restore soil quality and microbiota and control bacterial wilt disease. This study found new biomaterial to control soilborne bacterial disease by combining fulvic acid and poly-gamma-glutamic acid application.

Insights into the Biocontrol Function of a Burkholderia gladioli Strain against Botrytis cinerea.

Pathogenic fungi are the main cause of yield loss and postharvest loss of crops. In recent years, some antifungal microorganisms have been exploited and applied to prevent and control pathogenic fungi. In this study, an antagonistic bacteria KRS027 isolated from the soil rhizosphere of a healthy cotton plant from an infected field was identified as Burkholderia gladioli by morphological identification, multilocus sequence analysis, and typing (MLSA-MLST) and physiobiochemical examinations. KRS027 showed broad spectrum antifungal activity against various phytopathogenic fungi by secreting soluble and volatile compounds. KRS027 also has the characteristics of plant growth promotion (PGP) including nitrogen fixation, phosphate, and potassium solubilization, production of siderophores, and various enzymes. KRS027 is not only proven safe by inoculation of tobacco leaves and hemolysis test but also could effectively protect tobacco and table grapes against gray mold disease caused by Botrytis cinerea. Furthermore, KRS027 can trigger plant immunity by inducing systemic resistance (ISR) activated by salicylic acid- (SA), jasmonic acid- (JA), and ethylene (ET)-dependent signaling pathways. The extracellular metabolites and volatile organic compounds (VOCs) of KRS027 affected the colony extension and hyphal development by downregulation of melanin biosynthesis and upregulation of vesicle transport, G protein subunit 1, mitochondrial oxidative phosphorylation, disturbance of autophagy process, and degrading the cell wall of B. cinerea. These results demonstrated that B. gladioli KRS027 would likely become a promising biocontrol and biofertilizer agent against fungal diseases, including B. cinerea, and would promote plant growth. IMPORTANCE Searching the economical, eco-friendly and efficient biological control measures is the key to protecting crops from pathogenic fungi. The species of Burkholderia genus are widespread in the natural environment, of which nonpathogenic members have been reported to have great potential for biological control agents and biofertilizers for agricultural application. Burkholderia gladioli strains, however, need more study and application in the control of pathogenic fungi, plant growth promotion, and induced systemic resistance (ISR). In this study, we found that a B. gladioli strain KRS027 has broad spectrum antifungal activity, especially in suppressing the incidence of gray mold disease caused by Botrytis cinerea, and can stimulate plant immunity response via ISR activated by salicylic acid- (SA), jasmonic acid- (JA), and ethylene (ET)-dependent signaling pathways. These results indicate that B. gladioli KRS027 may be a promising biocontrol and biofertilizer microorganism resource in agricultural applications.

Enhancing bioefficacy of Bacillus amyloliquefaciens SF14 with salicylic acid for the control of the postharvest citrus green mould

Penicillium digitatum is a major post-harvest pathogen of citrus fruit causing tremendous economic losses. In this study, salicylic acid (SA) and antagonistic bacteria Bacillus amyloliquefaciens (SF14) were tested in controlling P. digitatum (green mould) infections as an alternative to chemical controls. Three concentrations of SA (0.5, 1, and 2%) were tested for their antifungal activity in both in vitro and in vivo conditions alone and in combination with SF14. Fruit quality parameters including fruit firmness, weight loss, titratable acidity, and soluble solids content were evaluated. The impact of these biological treatments was also investigated by measuring the changes in organic and mineral properties of orange fruits using Fourier transform infrared spectroscopy (FT-IR). Enzymatic activity including phenylalanine ammonia-lyase (PAL), total polyphenols and flavonoids was also tested. In vitro assays showed that SF14 combined with 0.5% SA provided the highest inhibition of green mould growth reaching 72.98%. Inhibition of spore germination was also highest in the same treatment od reaching 63.40%. FTIR of fungal biomass confirmed an alteration of fungal well wall following biological treatments. Moreover, during in vivo assays, the combination of SF14 and 0.5% SA produced the lowest disease severity (5.43%) without considerably affecting the fruit quality. Further investigation of the effect of these treatments on the mineral composition of treated oranges showed that Ca, Fe, K, Mg, and Na concentrations were significantly affected. Changes in these elements may show a defence reaction induced by the biological treatments against the pathogen, which was also confirmed with the measurement of PAL activity, total polyphenols, and flavonoids. Interestingly, FTIR of citrus peel indicated significant changes in the content of bands denoting for phenolic and aromatic rings, suggesting an induced resistance against the pathogenic fungus. Overall, our findings suggested that the combination of both bacterial antagonist SF14 and 0.5% SA can control the green mould of citrus, and as such constitutes a safer alternative to chemicals during postharvest storage.

Biological potential of Bacillus subtilis BS45 to inhibit the growth of Fusarium graminearum through oxidative damage and perturbing related protein synthesis.

Fusarium root rot (FRR) caused by Fusarium graminearum poses a threat to global food security. Biological control is a promising control strategy for FRR. In this study, antagonistic bacteria were obtained using an in-vitro dual culture bioassay with F. graminearum. Molecular identification of the bacteria based on the 16S rDNA gene and whole genome revealed that the species belonged to the genus Bacillus. We evaluated the strain BS45 for its mechanism against phytopathogenic fungi and its biocontrol potential against FRR caused by F. graminearum. A methanol extract of BS45 caused swelling of the hyphal cells and the inhibition of conidial germination. The cell membrane was damaged and the macromolecular material leaked out of cells. In addition, the mycelial reactive oxygen species level increased, mitochondrial membrane potential decreased, oxidative stress-related gene expression level increased and oxygen-scavenging enzyme activity changed. In conclusion, the methanol extract of BS45 induced hyphal cell death through oxidative damage. A transcriptome analysis showed that differentially expressed genes were significantly enriched in ribosome function and various amino acid transport pathways, and the protein contents in cells were affected by the methanol extract of BS45, indicating that it interfered with mycelial protein synthesis. In terms of biocontrol capacity, the biomass of wheat seedlings treated with the bacteria increased, and the BS45 strain significantly inhibited the incidence of FRR disease in greenhouse tests. Therefore, strain BS45 and its metabolites are promising candidates for the biological control of F. graminearum and its related root rot diseases.

A Field Evaluation of Sodium Silicate and Bacillus subtilis on the Growth and Yield of Bananas following Fusarium Wilt Disease Infection

A field investigation was conducted in a Fusarium-infected area to evaluate the effectiveness of silicate compounds with antagonist bacteria on morpho-physiological growth performance in bananas. The roots of banana plants were treated by drenching the soil with four different treatments: control (without any treatment), CBZ (Carbendazim fungicide alone), SS + BS (integration sodium silicate with Bacillus subtilis), and CBZ + SS + BS (integration CBZ, SS, and BS). All treatments were arranged in a randomized complete block design with four replications. The results confirm that plant height, pseudo-stem diameter, and canopy diameter markedly increased from weeks 2 to 10 after transplantation. Amendment with these elements induced a higher total chlorophyll content, which contributed to the increased rate of leaf gas exchange and biochemical changes for controlling Fusarium wilt disease infection. From these findings, the CBZ + SS + BS application in the farm had significantly reduced disease incidence by 16.07% and disease severity by 14.28%. The same treatment achieved the greatest disease reduction by 63.05%. Therefore, the integration between CBZ + SS + BS had good significant effects in controlling Fusarium wilt disease and enhanced the morpho-physiological growth performance with an average yield production of about 24.72 kg per fruit bunch.