Disease Control Efficacy Research Articles

Biocontrol potential of Bevibacillus brevis HK544 for fungal plant diseases

Chemical fungicides have been widely used to control plant diseases caused by fungal pathogens. Still, the overuse of these fungicides has led to concerns about the hazards to humans, animals, and the environment and an increase in fungicide resistance. Interest in biocontrol agents with strong antimicrobial activity has significantly increased to counteract the escalating risks of chemical fungicides. Our previous study found that a soil-born microbe, Brevibacillus brevis HK544, exhibited a promising antifungal activity against a plant pathogen, Fusarium graminearum, with edeine B1 identified as the active compound. Herein, we extended our study to investigate the in vitro and in vivo antifungal spectrum of B. brevis HK544, including edeine B1, against various plant pathogenic fungi and evaluated the possibility that B. brevis HK544 could be used in combination with chemical fungicides to enhance the disease control efficacy. With an in vitro broad-spectrum antifungal activity, the B. brevis HK544 culture filtrate, culture broth, and metabolite edeine B1 exhibited a promising disease control efficacy against tomato gray mold and tomato late blight among the tested fungal diseases. Based on the in vitro antifungal activity against Botrytis cinerea, when edeine B1 was combined with various chemical fungicides, the highest average synergy score among the tested combinations was observed in the edeine B1 and benomyl combination. The co-application of the B. brevis HK544 culture filtrate and benomyl improved the disease control efficacy against gray mold disease, supporting the in vitro result. Taken together, our results show that B. brevis HK544 has potential as a biological control agent for controlling plant pathogenic fungi and also that B. brevis HK544 could be combined with chemical fungicides to enhance the efficacy of antifungal agents.

Intercropping wheat and appropriate nitrogen supply can alleviate faba bean wilt disease by reshaping soil microbial community structure

RationaleThe faba bean-wheat intercropping system is widely adopted to mitigate the incidence of faba bean wilt. The application of nitrogen fertilizers is imperative for promoting crop growth. Determining the optimal nitrogen dosage is crucial for enhancing the disease control efficacy of intercropping systems. So far, the impact of different nitrogen input management on soil microbial communities during the occurrence of faba bean diseases under intercropping conditions of faba bean and wheat has not been clear. ObjectiveTo explore the effect of different nitrogen application levels on the inter-root soil microbial community of faba bean in intercropping scenarios and its relationship with the incidence of faba bean wilt. Materials and methodsTwo planting modes, namely sole faba bean cultivation (M) and faba bean-wheat intercropping (I), were established in both field and pot experiments. Four nitrogen application levels (N0: 0 kg ha−1; N1: 45 kg ha−1; N2: 90 kg ha−1; and N3: 135 kg ha−1) were used to assess the incidence of wilt disease in faba bean. Furthermore, collect soil samples from the rhizosphere of faba bean plants for enzyme and microbial assays in the rhizosphere soil. ResultsThe application of nitrogen (N1, N2, and N3) reduced the incidence of faba bean wilt in the monocultures. Among the same N application levels, intercropping exhibited a significant decrease in wilt incidence compared with monocropping, with the lowest incidence observed at the N2 level. Additionally, the rhizosphere enzyme activity in faba bean was maximized under intercropping at the N2 level. Intercropping increased the richness and diversity of soil microorganisms, leading to an adjusted microbial community structure and the promotion of beneficial microorganism functions. Notably, nitrogen application played a pivotal role in changing the rhizosphere soil microbial community structure of faba bean. A reasonable nitrogen application rate (N2) can significantly improve the disease control effects of intercropping. ConclusionIntercropping with N2(90 kg ha−1) resulted in a significant increase in microbial diversity in faba bean soil. This led to an adaptable microbial structure, effectively mitigating the occurrence of faba bean wilt and ultimately promoting soil function and health.

Biological control of the shot-hole disease in flowering cherry tree using antimicrobial compounds produced by Bacillus velezensis 8–2

BackgroundEffective control of shot-hole disease in flowering cherries is challenging because of multiple causative pathogens (bacteria and fungi). Bacillus species are well-known for their ability to control plant pathogens; therefore, biological control potential of a Bacillus isolate, B. velezensis 8–2, against SH disease on flowering cherry trees was investigated.ResultsThis study revealed strong antimicrobial activity of Bacillus velezensis 8–2 against various plant pathogenic bacteria and fungi, particularly focusing on Xanthomonas arboricola pv. pruni (Xap) and Mycosphaerella cerasella (Mc), which cause shot-hole (SH) disease in flowering cherry trees. In vitro assays showed that the fermentation filtrate of B. velezensis 8–2 inhibited bacterial and fungal growth with minimum inhibitory concentrations of 1.25–10% and 2.5–10%, respectively. UPLC-Q–Orbitrap–MS analysis revealed that B. velezensis 8–2 produced antagonistic compounds, including polyketides (difficidin and oxydifficidin) and cyclic lipopeptides (iturin A, fengycin, and surfatin). To enhance antimicrobial activity, fermentation parameters for optimal production of two antibacterial and three antifungal compounds were investigated in a 5 L jar fermenter. By regulating the agitation speed to sustain the state of vegetative cells, the production period was extended by 20 h at 400 rpm, resulting in maximum yields of 86.6 μg/mL for difficidin and 150.0 μg/mL for oxydifficidin within a 72 h fermentation period. In a field trial, a 500-fold diluted 10% suspension concentrate formulation of B. velezensis 8–2 effectively inhibited the development of SH disease, demonstrating 66.6% disease control and a 90.2% disease symptoms reduction.ConclusionsThis is the first report to assess the disease control efficacy of B. velezensis for the biocontrol of SH disease in the field. These results suggest that the application of B. velezensis 8–2 could serve as a practical alternative for managing various bacterial and fungal diseases, including the management of SH disease in flowering cherry trees.Graphical

Synthesis and Evaluation of Copper Oleate Nanoparticles Against Citrus Anthracnose Caused by Colletotrichum gloeosporioides: In Situ Experiments, In Vitro Bioassays, and Field Trials

AbstractAnthracnose in citrus is a severe disease caused by Colletotrichum species. Synthetic fungicides have been used to combat this citrus disease; however, they may have adverse health effects on humans and cause harmful environmental impacts. During a search for environmentally friendly fungicides, we successfully prepared organic copper (copper (II) oleate) nanoparticles by using natural chitosan polymer as a coating agent and stabilizer and by combining an exchange reaction and in situ coating method. Organic copper nanoparticles coated with chitosan polymer (Cu‐oleate@CS NPs) with Cu contents ranged from 5.0 to 6.0 % by weight. The nanoparticles have an average particle size ranging from 50 nm to 60 nm (TEM), covered by a layer of polymer with a thickness of about 10 to 12 nm. The analytical results of zeta potential and dynamic solution size showed that the nanoparticles are uniformly dispersed (90–120 nm by DLS), with high stability (zeta potential=−61.7 mV) and low cytoxicity to Vero healthy cell line. Cu‐oleate@CS NPs significantly inhibited the mycelial growth of C. gloeosporioides in vitro at 500 and 1000 μg/mL. In field trials, the best antifungal efficacy of Cu‐oleate@CS NPs against citrus anthracnose on orange plants was found at a spray concentration of 0.33 % with a disease control efficacy of 54.18 % at 14 days after the second spraying. Our study results demonstrated a novel preparation of Cu‐oleate@CS NPs and suggested that it could be used as a promising nanoformulation to control citrus anthracnose in vitro and in vivo.

Uncovering the multifaceted properties of 6-pentyl-alpha-pyrone for control of plant pathogens.

Some volatile organic compounds (VOCs) produced by microorganisms have the ability to inhibit the growth and development of plant pathogens, induce the activation of plant defenses, and promote plant growth. Among them, 6-pentyl-alpha-pyrone (6-PP), a ketone produced by Trichoderma fungi, has emerged as a focal point of interest. 6-PP has been isolated and characterized from thirteen Trichoderma species and is the main VOC produced, often accounting for >50% of the total VOCs emitted. This review examines abiotic and biotic interactions regulating the production of 6-PP by Trichoderma, and the known effects of 6-PP on plant pathogens through direct and indirect mechanisms including induced systemic resistance. While there are many reports of 6-PP activity against plant pathogens, the vast majority have been from laboratory studies involving only 6-PP and the pathogen, rather than glasshouse or field studies including a host plant in the system. Biopesticides based on 6-PP may well provide an eco-friendly, sustainable management tool for future agricultural production. However, before this can happen, challenges including demonstrating disease control efficacy in the field, developing efficient delivery systems, and determining cost-effective application rates must be overcome before 6-PP's potential for pathogen control can be turned into reality.

Edeine B1 produced by Brevibacillus brevis reduces the virulence of a plant pathogenic fungus by inhibiting mitochondrial respiration.

Plant pathogenic fungi cause serious diseases, which result in the loss of crop yields and reduce the quality of crops worldwide. To counteract the escalating risks of chemical fungicides, interest in biological control agents to manage plant diseases has significantly increased. In this study, we comprehensively screened microbial culture filtrates using a yeast screening system to find microbes exhibiting respiratory inhibition activity. Consequently, we found a soil-borne microbe Brevibacillus brevis HK544 strain exhibiting a respiration inhibitory activity and identified edeine B1 (EB1) from the culture filtrate of HK544 as the active compound of the respiration inhibition activity. Furthermore, against a plant pathogenic fungus Fusarium graminearum, our results showed that EB1 has effects on multiple aspects of respiration with the downregulation of most of the mitochondrial-related genes based on transcriptome analysis, differential EB1-sensitivity from targeted mutagenesis, and the synergistic effects of EB1 with electron transport chain complex inhibitors. With the promising plant disease control efficacy of B. brevis HK544 producing EB1, our results suggest that B. brevis HK544 has potential as a biocontrol agent for Fusarium head blight.IMPORTANCEAs a necrotrophic fungus, Fusarium graminearum is a highly destructive pathogen causing severe diseases in cereal crops and mycotoxin contamination in grains. Although chemical control is considered the primary approach to control plant disease caused by F. graminearum, fungicide-resistant strains have been detected in the field after long-term continuous application of fungicides. Moreover, applying chemical fungicides that trigger mycotoxin biosynthesis is a great concern for many researchers. Biocontrol of Fusarium head blight (FHB) by biological control agents (BCAs) represents an alternative approach and could be used as part of the integrated management of FHB and mycotoxin production. The most extensive studies on bacterial BCAs-fungal communications in agroecosystems have focused on antibiosis. Although many BCAs in agricultural ecology have already been used for fungal disease control, the molecular mechanisms of antibiotics produced by BCAs remain to be elucidated. Here, we found a potential BCA (Brevibacillus brevis HK544) with a strong antifungal activity based on the respiration inhibition activity with its active compound edeine B1 (EB1). Furthermore, our results showed that EB1 secreted by HK544 suppresses the expression of the mitochondria-related genes of F. graminearum, subsequently suppressing fungal development and the virulence of F. graminearum. In addition, EB1 exhibited a synergism with complex I inhibitors such as rotenone and fenazaquin. Our work extends our understanding of how B. brevis HK544 exhibits antifungal activity and suggests that the B. brevis HK544 strain could be a valuable source for developing new crop protectants to control F. graminearum.

Essential Oils for Managing Anthracnose in Mango (Mangifera indica): Laboratory Results Do Not Translate into Field Efficacy.

Essential oil-based products with broad plant disease control claims are commercially available and may be a practical alternative to copper fungicides for crop protection in organic mango orchards. We evaluated the disease control efficacy and crop safety of thyme oil, savory oil, and tree tea oil through replicated in vitro, in vivo (detached leaf and potted trees), and field assays. Three Colletotrichum species associated with mango anthracnose were tested in vitro, whereas only C. siamense was used for in vivo assays. Within the range of concentrations tested in vitro (62.5 to 2,000 μl active ingredient [a.i.]/liter), thyme and savory oil displayed fungicidal activity, whereas no fungistatic or fungicidal activity was observed with tea tree oil. In the in vivo assays, none of the treatments based on a preventive application rate of thyme (1,150 μl a.i./liter), savory (2,000 μl a.i./liter), or tea tree oil (342 μl a.i./liter) were effective in preventing the development of anthracnose on wounded and artificially inoculated leaves. Although field applications of thyme or tea tree oil did not result in phytotoxicity or negative impacts on fruit yield, they were ineffective in reducing the incidence and severity of naturally occurring anthracnose. Applications of copper hydroxide approved for organic agriculture were effective in controlling anthracnose in the field, and no added benefits were found by premixing this compound with thyme oil. Results indicate that essential oil products based on thyme or tea tree oil are inefficient at controlling anthracnose in mangoes.

Multisite Field Evaluation of Bacteriophages for Fire Blight Management: Incorporation of Ultraviolet Radiation Protectants and Impact on the Apple Flower Microbiome.

Fire blight, a disease of pome fruits caused by the bacterium Erwinia amylovora, has become increasingly difficult to manage after the emergence of streptomycin-resistant strains. Alternative antibiotics and copper are available; however, these chemicals have use restrictions in some countries and also can carry risks of phytotoxicity. Therefore, there is growing interest in biological-based management options, with bacteriophage (phages) showing promise, as these naturally occurring pathogens of bacteria are easy to isolate and grow. However, there are several technical challenges regarding the implementation of phage biocontrol in the field, as the viral molecules suffer from ultraviolet radiation (UVR) degradation and can die off rapidly in the absence of the host bacterium. In this work, we assessed the efficacy of Erwinia phages and a commercial phage product for blossom blight control in the field across multiple locations in the eastern United States. In these tests, disease control ranged from 0.0 to 82.7%, and addition of a UVR protectant only resulted in significantly increased disease control in 2 of 12 tests. We also analyzed microbial community population changes in response to phage application. Changes in bacterial community diversity metrics over time were not detected; however, relative abundances of target taxa were temporarily reduced after phage applications, indicating that these phage applications did not have deleterious effects on the flower microbiome. We have demonstrated that biological control of fire blight with phages is achievable, but a better understanding of phage-pathogen dynamics is required to optimize disease control efficacy.

The rice endophytic bacterium Bacillus velezensis LS123N provides protection against multiple pathogens and enhances rice resistance to wind with increase in yield

Rice (Oryza sativa) is a globally important staple food, but its production was influenced by various biotic and abiotic stresses. This study involved screening endophytic bacterial strains for their antagonistic activities and control efficacy against multiple rice pathogens in planta assays. Subsequent greenhouse investigations identified Bacillus velezensis LS123N as the most promising strain for effectively controlling various rice diseases, including bacterial blight, rice blast, bakanae disease, Pythium disease, and brown spot disease. Field trials further demonstrated that LS123N significantly reduced naturally occurring brown spot disease caused by Bipolaris oryzae in both seedlings and mature rice plants, and increased rice yields. For disease control against bacterial blight, Pythium disease, and brown spot, only a single treatment during seed germination resulted in significant disease control efficacy. The application of LS123N during seed germination aligns with common rice cultivation practices. Additionally, LS123N exhibited the capacity to improve rice resistance to wind stress and enhance rice growth during the seedling stage. Endophytic colonization of LS123N was evidenced from seed to mature rice plants after seed treatment. Interestingly, the expression of many rice defense genes was suppressed within 48 h of the LS123N introduction. In vitro assays showed that LS123N inhibited spore germination and normal growth of germ tubes of B. oryzae, produced multiple hydrolases, siderophores, and IAA, and had the ability to solubilize phosphorus compounds. In summary, these results indicate that B. velezensis LS123N is a promising biocontrol agent and has the potential to be developed into a biofungicide, biofertilizer, and biostimulator.

Research Progress on Benzimidazole Fungicides: A Review.

Benzimidazole fungicides are a class of highly effective, low-toxicity, systemic broad-spectrum fungicides developed in the 1960s and 1970s, based on the fungicidal activity of the benzimidazole ring structure. They exhibit biological activities including anticancer, antibacterial, and antiparasitic effects. Due to their particularly outstanding antibacterial properties, they are widely used in agriculture to prevent and control various plant diseases caused by fungi. The main products of benzimidazole fungicides include benomyl, carbendazim, thiabendazole, albendazole, thiophanate, thiophanate-methyl, fuberidazole, methyl (1-{[(5-cyanopentyl)amino]carbonyl}-1H-benzimidazol-2-yl) carbamate, and carbendazim salicylate. This article mainly reviews the physicochemical properties, toxicological properties, disease control efficacy, and pesticide residue and detection technologies of the aforementioned nine benzimidazole fungicides and their main metabolite (2-aminobenzimidazole). On this basis, a brief outlook on the future research directions of benzimidazole fungicides is presented.

Genome Monitoring of SARS-CoV-2 Circulating in the Kyrgyz Republic in 2020–2021

Introduction: The RNA-containing severe acute respiratory syndrome virus SARS-CoV-2 has spread rapidly around the world by adaptation to the host through genetic evolution. The emergence of variants with genetic mutations that increase contagiousness and transmission may hamper the effectiveness of disease control programs and efficacy of vaccination. Objective: To establish dominant SARS-CoV-2 variants circulating in the Kyrgyz Republic in 2020–2021. Materials and methods: Genomic monitoring was carried out based on positive results of testing nasopharyngeal swabs. SARS-CoV-2 was detected by a real-time reverse transcription –polymerase chain reaction (RT-PCR) assay using registered commercial test kits. Genetic variants (n = 15) were determined by high-throughput sequencing on a MiSeq device (Illumina, USA) using the COVID-19 ARTIC v3 protocol. Mutational variability of SARS-CoV-2 was examined using a cluster analysis of amino acid substitutions in the S protein using Ward’s method. Nucleotide sequences from Kyrgyzstan (n = 15), Russia (n = 16), India (n = 2), and China (n = 2) were aligned using MAFFT. IQ-TREE v1.6.12 was used to infer the phylogenetic tree by maximum likelihood applying Nextstrain processes. Isolates Wuhan/Hu-1/2019 and Wuhan/WH01/2019 downloaded from the GenBank® database were considered to be the root of the tree (reference). Results: Phylogenetic data analysis revealed that SARS-CoV-2 B.1.1.7 (Alpha) was the dominant VOC variant, the proportion of which was as high as 36.4 % (12/33); B.1.351 (Beta) was also found (6.1 % or 2/33). When samples were examined for amino acid substitutions in the S-protein, B.1.1.7 Alpha (British) isolates were found to cluster into two distinct branches. Conclusion: The study of the frequency and influence of mutations on pathogenetic properties of the virus, as well as the analysis of the predominant variants of the virus will allow timely measures to be taken to counteract the spread of SARS-CoV-2 in the country. In this regard, continuous genome monitoring of circulating COVID-19 variants is necessary.

ONE SHOT - single shot radiotherapy for localized prostate cancer: 18-month results of a single arm, multicenter phase I/II trial

PurposeTo assess in a prospective, multicenter, single-arm phase I/II study the early safety and efficacy profile of single fraction urethra-sparing stereotactic body radiotherapy (SBRT) for men with localized prostate cancer. Material and methodsPatients with low- and intermediate-risk localized prostate cancer without significant tumor in the transitional zone were recruited. A single-fraction of 19 Gy was delivered to the prostate, with 17 Gy dose-reduction to the urethra. Intrafraction motion was monitored using intraprostatic electromagnetic transponders with intra-fraction correction of displacements exceeding 3 mm. Genitourinary (GU), gastrointestinal (GI), and sexual toxicity during the first 18 months were evaluated using the CTCAE v4.0 grading scale. Quality of life was assessed using the International Prostate Symptom Score, the Expanded Prostate Cancer Index composite 26 score, and the International Index of Erectile Function score. ResultsAmong the 45 patients recruited in 5 centers between 2017 and 2022, 43 received the single fraction without protocol deviations, and 34 had a minimal follow-up of 18 months. The worst GU toxicity was observed at day-5 after SBRT (42.5 % and 20 % with grade 1 and 2, respectively), returning to baseline at week-12 and month-6 (<3% with grade 2), with a 12 % grade 2 flare at month 18. Gl toxicity was mild in the acute phase, with no grade ≥ 2 events (12 % grade 1 at month 6). Grade-3 proctitis was observed in one patient at month 12, with < 3 % grade 2 toxicity at month 18. Mean GU and GI bother scores showed a decline at day 5, a complete recovery at month 6, and a flare between month 12 and 18. Mean PSA dropped from 6.2 ng/ml to 1.2 ng/ml at month 18 and 0.7 ng/ml at month 24. After a median follow-up time of 26 months, 3 biochemical failures (7 %) were observed at month 17, 21 and 30. ConclusionsIn this multicenter phase I/II trial, we demonstrated that a 19 Gy single-fraction urethra-sparing SBRT is feasible and associated with an acceptable toxicity rate, mostly returning to the baseline at week-12 and with a symptoms flare between months 12 and 18. Longer follow-up is needed to assess the potential long-term adverse effects and the disease control efficacy.

Fungicide strategies for Ramularia Leaf Spot control recommended in Uruguay and its residues in barley grains

Ramularia leaf spot (RLS) is primarily managed by foliar fungicide spraying, which can result in residues in the grain. In recent years, increasing attention has been paid to the risks these residues pose to consumers. This study aimed to evaluate the efficacy of fungicide management used for RLS control and their fungicide residues in barley grain. Four different alternatives of fungicide mixtures: fluxapyroxad + pyraclostrobin + epoxiconazole; pyraclostrobin + epoxiconazole + chlorothalonil; prothioconazole + trifloxystrobin, and isopyrazam + azoxystrobin, in three spray timings: GS33, GS47 and GS33+GS47, were evaluated in five field experiments. An untreated and a fully protected treatment were included. Fungicide residues, disease severity, control efficacy, area under the disease progress curve, and plump grain yield were calculated. All fungicide strategies adhered to food safety, complying with the Maximum Residue Limits established by Codex and the European Union. Effectiveness varied based on RLS development, application time, and number of applications. Fungicide treatments applied at GS33+GS47 were the most effective as the highest severity levels were observed at the stage after GS47 under the conditions studied. Plump grain yield only showed minimal differences in the late epidemic. The study emphasized the low risk of fungicide presence in grains, favoring efficacy when selecting RLS management options. Any changes in management or regulations should be carefully reviewed to maintain findings. The research underscored the compatibility of recommended fungicide treatments with food safety standards, highlighting the balance between disease control efficacy and consumer safety.

Co-application of Brassinolide and Pyraclostrobin Improved Disease Control Efficacy by Eliciting Plant Innate Defense Responses in Arabidopsis thaliana.

Applying brassinolide (BL, a phytohormone) in combination with pyraclostrobin (Pyr, a fungicide) has shown effective disease control in field trials. However, the mechanism by which BL + Pyr control disease remains uncertain. This work compared the disease control and defense responses of three pretreatments (BL, Pyr, and BL + Pyr) in Arabidopsis thaliana. We found that BL + Pyr improved control against Pyr-sensitive Hyaloperonospora arabidopsidis and Botrytis cinerea by 19 and 17% over Pyr, respectively, and achieved 29% control against Pyr-resistant B. cinerea. Furthermore, BL + Pyr outperformed BL or Pyr in boosting transient H2O2 accumulation, and the activities of POD, APX, GST, and GPX. RNA-seq analysis revealed a more potent activation of defense genes elicited by BL + Pyr than by BL or Pyr. Overall, BL + Pyr controlled disease by integrating the elicitation of plant innate disease resistance with the fungicidal activity of Pyr.

Infection Risk-Based Application of Plant Resistance Inducers for the Control of Downy and Powdery Mildews in Vineyards

Plant resistance inducers (PRIs) are potential alternatives for controlling grapevine downy (DM) and powdery (PM) mildews in vineyards. In a 3-year field study, we evaluated the field efficacy of six commercial PRIs of chemical and natural origin against DM and PM diseases when applied at designated vine growth stages in a mixture with low doses of copper and sulfur, and only when advised by weather-driven disease models. The disease severity and incidence were evaluated for each season at key growth stages (i.e., the end of flowering, berries pea-sized, veraison, and pre-harvest), and areas under the disease progress curves (AUDPC) were calculated and compared with those of nontreated vines. These risk-based applications resulted in a 41% and 61% reduction of interventions against DM and PM, respectively, compared to the official advice for integrated pest management in the growing area. These applications provided a disease control efficacy of 88% for DM and 93% for PM; the disease severity on bunches never exceeded 5%. Overall, when the disease severity was expressed as AUDPC, we observed higher efficacy of all the PRIs for PM, and of laminarin and cerevisane for DM. We also found that potassium phosphonate and fosetyl-Al (commonly used against DM) were effective against PM, and cos-oga (used against PM) was effective against DM. These results broaden the application and integration of PRIs in viticulture.

Pentaene macrolides AB023a and takanawaene C produced by Streptomyces xanthocidicus strain S3 for controlling pepper anthracnose

A natural product library consisting of the culture extracts of 814 actinomycete strains was screened for antifungal compounds that disrupt the cell integrity of plant pathogenic fungi using an adenylate kinase (AK) assay system. The culture extract of Streptomyces xanthocidicus strain S3 exhibited high AK activity against various plant pathogens. The active ingredients, AT-1 and AT-2, were isolated from the culture extract using a series of chromatographic procedures. Based on MS, UV, and NMR spectrometric analyses, the structures of AT-1 and AT-2 were determined as the pentaene macrolides, AB023a and takanawaene C. AB023a and takanawaene C displayed broad-spectrum antifungal activity against Aspergillus oryzae, Botrytis cinerea, Colletotrichum coccodes, C. gloeosporioides, C. orbiculare, Cylindrocarpon destructans, and Fusarium oxysporum f. sp. lycopersici, showing minimum inhibitory concentrations of 1–32 μg/mL. Treatment of AB023a and takanawaene C successfully inhibited anthracnose development on pepper plants in a concentration-dependent manner without phytotoxicity. The disease control efficacy of both compounds was comparable to that of the commercial fungicide chlorothalonil. Collectively, these results suggest that the polyene macrolides produced by S. xanthocidicus strain S3 can be used as natural fungicides for plant disease control.

Impact of bacterial synthesized nanoparticles on quality attributes and postharvest disease control efficacy of apricot and loquat.

Postharvest fungal attacks on fruits such as apricots and loquats are common. Diseased fruit samples were collected from Murree's local fruit markets. The disease-causing pathogens were identified utilizing molecular, microscopic, and morphological characteristics. Alternaria alternata and Aspergillus niger were identified as the pathogens responsible for brown rot in loquat and black rot in apricot. To combat these fruit diseases, iron oxide (Fe2 O3 ) nanoparticles were synthesized using Bacillus subtilis and were characterized using various techniques. X-ray diffraction examination validated the size of iron oxide nanoparticles. The presence of several capping agents in the synthesized nanoparticles was confirmed by Fourier transform infrared analysis. Scanning electron microscopy revealed the spherical morphology of nanoparticles, whereas energy-dispersive X-ray proved the presence of different elemental compositions. After completing antifungal activities in vitro and in vivo, it was discovered that a nanoparticle concentration of 1.0mg/mL efficiently suppressed the growth of fungal mycelia. Fungi growth was effectively inhibited in fruit samples treated with 1.0mg/mL nanoparticles. The results of successful in vitro and in vivo antifungal activities imply that iron oxide (Fe2 O3 ) nanoparticles play an important role in ensuring fruit quality against pathogenic attacks. Bacterial-mediated iron oxide can be widely used because it is less expensive and less harmful to the environment than chemically manufactured fertilizers.

In vitro and in vivo antimicrobial activity of the fungal metabolite toluquinol against phytopathogenic bacteria.

Bacterial plant diseases cause tremendous economic losses worldwide. However, a few effective and sustainable control methods are currently available. To discover novel and effective management approaches, we screened marine fungi for their antibacterial activity against phytopathogenic bacteria in vitro and in vivo. We screened the culture broth of 55 fungal strains isolated from various marine sources (seawater, algae, and sediment) for their in vitro antibacterial activity using the broth microdilution method. Then, only the fungal strain (designated UL-Ce9) with higher antibacterial activity in vitro was tested in an in vivo experiment against tomato bacterial wilt. The active compounds of UL-Ce9 were extracted using ethyl acetate, purified by a series of chromatography, and the structure was elucidated by nuclear magnetic resonance spectroscopy. Pesticide formulations of toluquinol were prepared as soluble concentrates and wettable powder. The disease control efficacy of toluquinol formulations was evaluated against blight of rice and the bacterial wilt of tomato. The culture broth of UL-Ce9 showed high antibacterial activity against Agrobacterium tumefaciens, Ralstonia solanacearum, and Xanthomonas arboricola pv. pruni in vitro, and we selected UL-Ce9 for the in vivo test. The UL-Ce9 culture broth completely suppressed the bacterial wilt of tomato at a dilution of 1:5. The phylogenetic analysis identified UL-Ce9 as Penicillium griseofulvum, and the antibacterial metabolites were revealed as patulin, gentisyl alcohol, and toluquinol, all of which were associated with the biosynthetic pathway of the mycotoxin patulin. Patulin exhibited the highest antibacterial activity against 16 phytopathogenic bacteria in vitro, followed by toluquinol and gentisyl alcohol. As patulin is toxic, we selected toluquinol to investigate its potential use as a pesticide against bacterial plant diseases. Compared with the chemicals currently being applied in agriculture (streptomycin and oxytetracycline), toluquinol formulations exhibited similar and higher control efficacies against bacterial leaf blight of rice and bacterial wilt of tomato, respectively. To the best of our knowledge, this is the first report of the antibacterial activity of toluquinol against phytopathogenic bacteria. Our results suggest that toluquinol is a potential candidate for the development of novel and effective pesticides for the management of bacterial plant diseases.

Baseline sensitivity of Botrytis cinerea strawberry isolates from Brazil to fludioxonil and cyprodinil and disease control efficacy

Baseline sensitivity of Botrytis cinerea strawberry isolates from Brazil to fludioxonil and cyprodinil and disease control efficacy

Lipopeptide-enriched extracts of Bacillus velezensis B157 for controlling tomato early blight

Bacillus strains used in plant disease control produce cyclic lipopeptides (LPs) with antifungal activities. The LP concentration in Bacillus-cell suspensions may be insufficient to protect aerial plant parts against phytopathogens and the de novo metabolite production by bacteria on leaf surfaces is limited, leading to inconsistent control results in the field. We investigated the use of LP-enriched crude organic extracts of Bacillus velezensis strain B157 for controlling early blight, a destructive tomato disease caused by Alternaria linariae. Liquid chromatography–tandem mass spectrometry-based LP dereplication and molecular networking analysis revealed a complex mixture of iturins, fengycins, and surfactins in the organic extract of B157. Fengycins and iturins showed a marked in vitro antifungal activity. Treating tomato plants with the organic extract significantly reduced the severity of early blight, attaining more than 90% of disease control efficacy in two independent greenhouse experiments. These results show the potential of using LP-enriched biochemical fungicides for plant disease control.