ABSTRACT
Rhizobacteria play a crucial role in plant health by providing natural antagonism against soil-borne fungi. The use of rhizobacteria has been viewed as an alternative to the use of chemicals that could be useful for the integrated management of plant diseases and also increase yield in an environmentally friendly manner. However, there is limited understanding of the specific mechanisms by which rhizobacteria inhibit these pathogens and the diversity of rhizobacterial species involved. This study aims to isolate, identify, and characterize rhizobacteria with antagonistic activities against soil-borne fungi. Laboratory tests were carried out on isolated rhizobacteria to evaluate their inhibitory activity against Rhizoctonia solani, Pythium aphanidermatum and Macrophomina phaseolina. The selected bacteria were identified using the Vitek 2 compact system and 16S rRNA genes. Experiments were carried out to evaluate the plant growth promotion and biocontrol ability of these selected isolates. Out of 324 rhizobacteria isolates obtained from various plant species, twelve were chosen due to their strong (>50 %) wide-ranging antifungal activity against three significant phytopathogenic fungi species. According to the identification results, they belong to the following species: Aeribacillus pallidus ECC4, Alloiococcus otitis BRE6, Aneurinibacillus thermoaerophilus ECL1, A. thermoaerophilus SDV1, Bacillus halotolerans DMC8, B. megaterium SKE2, B. megaterium TNK1, B. subtilis NAS1, Enterobacter cloacae complex BZD3, Leclercia adecarboxylata DKS3, Paenibacillus polymyxa TRS4, and Staphylococcus lentus BZD2. Eleven isolates produced protease, six isolates produced chitinase, and seven isolates were highly effective in producing hydrogen cyanide. Ten isolates could fix nitrogen, while all isolates could produce potassium, indole-3-acetic acid, siderophore, and ammonia. These findings enhance our understanding of rhizobacterial biodiversity and their potential as biocontrol agents in sustainable agriculture.
ABSTRACT
Staphylococcus aureus, a major foodborne pathogen, is frequently detected in fresh produce. It often causes food poisoning accompanied by abdominal pain, diarrhea, and vomiting. Additionally, the abuse of antibiotics to control S. aureus has resulted in the emergence of antibiotics-resistant bacteria, such as methicillin resistant S. aureus. Therefore, bacteriophage, a natural antimicrobial agent, has been suggested as an alternative to antibiotics. In this study, a lytic phage SSP49 that specifically infects S. aureus was isolated from a sewage sample, and its morphological, biological, and genetic characteristics were determined. We found that phage SSP49 belongs to the Straboviridae family (Caudoviricetes class) and maintained host growth inhibition for 30 h in vitro. In addition, it showed high host specificity and a broad host range against various S. aureus strains. Receptor analysis revealed that phage SSP49 utilized cell wall teichoic acid as a host receptor. Whole genome sequencing revealed that the genome size of SSP49 was 137,283 bp and it contained 191 open reading frames. The genome of phage SSP49 did not contain genes related to lysogen formation, bacterial toxicity, and antibiotic resistance, suggesting its safety in food application. The activity of phage SSP49 was considerably stable under various high temperature and pH conditions. Furthermore, phage SSP49 effectively inhibited S. aureus growth on baby spinach leaves both at 4 °C and 25 °C while maintaining the numbers of active phage during treatments (reductions of 1.2 and 2.1 log CFU/cm2, respectively). Thus, this study demonstrated the potential of phage SSP49 as an alternative natural biocontrol agent against S. aureus contamination in fresh produce.
Subject(s)
Host Specificity , Plant Leaves , Spinacia oleracea , Staphylococcus aureus , Spinacia oleracea/microbiology , Staphylococcus aureus/virology , Plant Leaves/microbiology , Food Microbiology , Genome, Viral , Bacteriophages/isolation & purification , Bacteriophages/physiology , Food Contamination/prevention & control , Staphylococcus Phages , Whole Genome Sequencing , Sewage/virology , Sewage/microbiologyABSTRACT
Introduction: Stem and crown gall disease caused by the plant pathogen Rhizobium radiobacter has a significant impact on highbush blueberry (Vaccinium corymbosum) production. Current methods for controlling the bacterium are limited. Lytic phages, which can specifically target host bacteria, have been widely gained interest in agriculture. Methods: In this study, 76 bacteriophages were recovered from sewage influent and screened for their inhibitory effect against Rhizobium spp. The phages were genetically characterized through whole-genome sequencing, and their lytic cycle was confirmed. Results: Five potential candidate phages (isolates IC12, IG49, AN01, LG08, and LG11) with the ability to lyse a broad range of hosts were chosen and assessed for their morphology, environmental stability, latent period, and burst size. The morphology of these selected phages revealed a long contractile tail under transmission electron microscopy. Single-step growth curves displayed that these phages had a latent period of 80-110 min and a burst size ranging from 8 to 33 phages per infected cell. None of these phages contained any antimicrobial resistance or virulence genes in their genomes. Subsequently, a combination of two-, three- and four-phage cocktails were formulated and tested for their efficacy in a broth system. A three-phage cocktail composed of the isolates IC12, IG49 and LG08 showed promising results in controlling a large number of R. radiobacter strains in vitro. In a soil/peat-based model, the three-phage cocktail was tested against R. radiobacter PL17, resulting in a significant reduction (p < 0.05) of 2.9 and 1.3 log10 CFU/g after 24 and 48 h of incubation, respectively. Discussion: These findings suggest that the three-phage cocktail (IC12, IG49 and LG08) has the potential to serve as a proactive antimicrobial solution for controlling R. radiobacter on blueberry.
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The management of an alien Rugose spiraling whitefly (RSW) on oil palm using a native entomopathogenic fungus, Isaria fumosorosea pfu5 necessitated community approach for pest management. Moreover, coverage of huge leaf biomass warrants massive multiplication of biocontrol agent. In this communication, a two-step strategy, first including pure culture production and the second including ready-to-use culture production of the biocontrol agent is disclosed. The production costs and success of this technology in RSW management of oil palm are also discussed.
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This study investigates the efficacy of Trichoderma spp. and Bacillus spp., as well as their gamma radiation-induced mutants, as potential biological control agents against Meloidogyne javanica (Mj) in tomato plants. The research encompasses in vitro assays, greenhouse trials, and molecular identification methodologies to comprehensively evaluate the biocontrol potential of these agents. In vitro assessments reveal significant nematicidal activity, with Bacillus spp. demonstrating notable effectiveness in inhibiting nematode egg hatching (16-45%) and inducing second-stage juvenile (J2) mortality (30-46%). Greenhouse trials further confirm the efficacy of mutant isolates, particularly when combined with chitosan, in reducing nematode-induced damage to tomato plants. The combination of mutant isolates with chitosan reduces the reproduction factor (RF) of root-knot nematodes by 94%. By optimizing soil infection conditions with nematodes and modifying the application of the effective compound, the RF of nematodes decreases by 65-76%. Molecular identification identifies B. velezensis and T. harzianum as promising candidates, exhibiting significant nematicidal activity. Overall, the study underscores the potential of combined biocontrol approaches for nematode management in agricultural settings. However, further research is essential to evaluate practical applications and long-term efficacy. These findings contribute to the development of sustainable alternatives to chemical nematicides, with potential implications for agricultural practices and crop protection strategies.
Subject(s)
Bacillus , Gamma Rays , Pest Control, Biological , Plant Diseases , Solanum lycopersicum , Tylenchoidea , Animals , Tylenchoidea/physiology , Bacillus/genetics , Bacillus/physiology , Solanum lycopersicum/parasitology , Solanum lycopersicum/microbiology , Plant Diseases/parasitology , Plant Diseases/prevention & control , Plant Diseases/microbiology , Pest Control, Biological/methods , Mutation , Hypocreales/genetics , Antinematodal Agents/pharmacology , Biological Control Agents/pharmacology , Chitosan/pharmacologyABSTRACT
This study demonstrates that root-associated Kosakonia oryziphila NP19, isolated from rice roots, is a promising plant growth-promoting bioagent and biopesticide for combating rice blast caused by Pyricularia oryzae. In vitro experiments were conducted on fresh leaves of Khao Dawk Mali 105 (KDML105) jasmine rice seedlings. The results showed that NP19 effectively inhibited the germination of P. oryzae fungal conidia. Fungal infection was suppressed across three different treatment conditions: rice colonized with NP19 and inoculated by fungal conidia, a mix of NP19 and fungal conidia concurrently inoculated on the leaves, and fungal conidia inoculation first followed by NP19 inoculation after 30 h. Additionally, NP19 reduced fungal mycelial growth by 9.9-53.4%. In pot experiments, NP19 enhanced the activities of peroxidase (POD) and superoxide dismutase (SOD) by 6.1-63.0% and 3.0-67.7%, respectively, indicating a boost in the plant's defense mechanisms. Compared to the uncolonized control, the NP19-colonized rice had 0.3-24.7% more pigment contents, 4.1% more filled grains per panicle, 26.3% greater filled grain yield, 34.4% higher harvest index, and 10.1% more content of the aroma compound 2-acetyl-1-pyrroline (2AP); for rice colonized with NP19 and infected with P. oryzae, these increases were 0.2-49.2%, 4.6%, 9.1%, 54.4%, and 7.5%, respectively. In field experiments, blast-infected rice that was colonized and/or inoculated with NP19 treatments had 15.1-27.2% more filled grains per panicle, 103.6-119.8% greater filled grain yield, and 18.0-35.8% higher 2AP content. A higher SOD activity (6.9-29.5%) was also observed in the above-mentioned rice than in the blast-infected rice that was not colonized and inoculated with NP19. Following blast infection, NP19 applied to leaves decreased blast lesion progression. Therefore, K. oryziphila NP19 was demonstrated to be a potential candidate for use as a plant growth-promoting bioagent and biopesticide for suppressing rice blast.
Subject(s)
Oryza , Plant Diseases , Oryza/microbiology , Oryza/growth & development , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Roots/microbiology , Plant Roots/growth & development , Spores, Fungal , Plant Leaves/microbiology , Ascomycota/pathogenicity , Seedlings/microbiology , Seedlings/growth & development , Biological Control Agents/pharmacology , Peroxidase/metabolismABSTRACT
The escalating interest in Bacillus velezensis as a biocontrol agent arises from its demonstrated efficacy in inhibiting both phytopathogenic fungi and bacteria, positioning it as a promising candidate for biotechnological applications. This mini review aims to offer a comprehensive exploration of the multifaceted properties of B. velezensis, with particular focus on its beneficial interactions with plants and its potential for controlling phytopathogenic fungi. The molecular dialogues involving B. velezensis, plants, and phytopathogens are scrutinized to underscore the intricate mechanisms orchestrating these interactions. Additionally, the review elucidates the mode of action of B. velezensis, particularly through cyclic lipopeptides, highlighting their importance in biocontrol and promoting plant growth. The agricultural applications of B. velezensis are detailed, showcasing its role in enhancing crop health and productivity while reducing reliance on chemical pesticides. Furthermore, the review extends its purview in the industrial and environmental arenas, highlighting its versatility across various sectors. By addressing challenges such as formulation optimization and regulatory frameworks, the review aims to chart a course for the effective utilization of B. velezensis. KEY POINTS: ⢠B. velezensis fights phytopathogens, boosting biotech potential ⢠B. velezensis shapes agri-biotech future, offers sustainable solutions ⢠Explores plant-B. velezensis dialogue, lipopeptide potential showcased.
Subject(s)
Bacillus , Plant Diseases , Bacillus/metabolism , Plant Diseases/microbiology , Plant Diseases/prevention & control , Biological Control Agents/metabolism , Biological Control Agents/pharmacology , Lipopeptides/metabolism , Lipopeptides/pharmacology , Fungi/metabolism , Fungi/drug effects , Plant Development , Pest Control, Biological/methods , Biotechnology/methods , Crops, Agricultural/microbiology , Plants/microbiology , Agriculture/methodsABSTRACT
Bioactive secondary metabolites from fungi, including Trichoderma, are an excellent source of plant biostimulants. Although production of novel biostimulants from known microbes is critical, challenging them may produce novel bioactive compounds. With this hypothesis, the study used live Fusarium chlamydosporum (FOL7) culture as the inducer during T. harzianum (IF63) growth in broth. Plate assays and gas chromatography-mass spectrometry (GC-MS) analysis were used to characterise the metabolites. Microscopy, pot experiments and, biochemical estimations of the defence-related enzymes in tomato plants established the biostimulant activity of the induced Trichoderma metabolites. Fungal crude metabolites (FCM) obtained from IF63+FOL7 extracts (TF.ex) showed increased antimicrobial activity. TF.ex at 50 µg mL-1, inhibited the FOL7 growth by 68.33% compared to the Trichoderma alone extract. Scanning electron microscopy (SEM) revealed morphological disruption of FOL7 mycelia by TF.ex. GC-MS analysis of the extracts revealed the presence of approximately 64 compounds, of which at least 13 were detected explicitly in TF.ex. Methyl (3-oxo-2-pentylcyclopentyl) acetate (Methyl dihydrojasmonate), a lipid functionally related to jasmonic acid, was the major metabolite (â¼21%) present in TF.ex. Tomato seed dressing with TF.ex promoted plant growth and induced systemic resistance against FOL7 compared to alone Trichoderma and Fusarium extracts. The TF.ex treatment increased the superoxide dismutase (33%) and catalase activity by 2.5-fold in tomato plants. The study concludes that fungal secondary metabolites may be modulated by providing appropriate challenges to produce effective metabolite-based biostimulants for agricultural applications.
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In the prospect of novel potential biocontrol agents, a new strain BDI-IS1 belonging to the recently described Bacillus nakamurai was selected for its strong in vitro antimicrobial activities against a range of bacterial and fungal phytopathogens. Genome mining coupled with metabolomics revealed that BDI-IS1 produces multiple non-ribosomal secondary metabolites including surfactin, iturin A, bacillaene, bacillibactin and bacilysin, together with some some ribosomally-synthesized and post-translationally modified peptides (RiPPs) such as plantazolicin, and potentially amylocyclicin, bacinapeptin and LCI. Reverse genetics further showed the specific involvement of some of these compounds in the antagonistic activity of the strain. Comparative genomics between the five already sequenced B. nakamurai strains showed that non-ribosomal products constitute the core metabolome of the species while RiPPs are more strain-specific. Although the secondary metabolome lacks some key bioactive metabolites found in B. velezensis, greenhouse experiments show that B. nakamurai BDI-IS1 is able to protect tomato and maize plants against early blight and northern leaf blight caused by Alternaria solani and Exserohilum turcicum, respectively, at levels similar to or better than B. velezensis QST713. The reduction of these foliar diseases, following root or leaf application of the bacterial suspension demonstrates that BDI-IS1 can act by direct antibiosis and by inducing plant defence mechanisms. These findings indicate that B. nakamurai BDI-IS1 can be considered as a good candidate for biocontrol of plant diseases prevailing in tropical regions, and encourage further research into its spectrum of activity, its requirements and the conditions needed to ensure its efficacy.
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Tea red spider mite (TRSM), Oligonychus coffeae Nietner, is one of the major pests that cause considerable crop losses in all tea-growing countries. TRSM management often involves the use of multiple chemical pesticides that are linked to human health risks and environmental pollution. Considering these critical issues, employing biocontrol agents is a potential green approach that may replace synthetic pesticides. This review study aims to discuss the efficacy of plant extracts, entomopathogenic microorganisms, and predators in controlling TRSM. This study includes 44 botanical extracts, 14 microbial species, and 8 potential predators used to control TRSM, along with their respective modes of action. Most of the botanical extracts have ovicidal, adulticidal, and larvicidal activity, ranging from 80 to 100 %, attributed to bioactive compounds such as phenols, alcohols, alkaloids, tannins, and other secondary metabolites. Among microbial pesticides, Purpureocillium lilacinum, Metarhizium robertsii, Aspergillus niger, Pseudomonas fluorescens, and Pseudomonas putida are highly effective against TRSM without causing any harm to the nontarget beneficial insects. Besides, some predators, including green lacewings, ladybirds, and phytoseiid mites have the potential to control TRSM. Employing these biocontrol agents simultaneously in tea plantations could be more effective in preventing TRSM. Nevertheless, their high biodegradability rate, uneven distribution, and uncontrolled release pose challenges for large-scale field applications. This study also explores how nanotechnology can enhance sustainability by addressing the limitations of biopesticides in field conditions. This review study could contribute to the search for potential biocontrol agents and the development of commercial nano biopesticides to control TRSM.
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Pseudomonas aeruginosa Y010, isolated from the taro rhizosphere, exhibits great antagonistic abilities against Dickeya strains that cause soft-rot and blackleg diseases of plants by producing potent antimicrobial agents. The complete genome of Y010 was sequenced and annotated, which is 6,415,628 bp in length with 66.39% GC content.
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Priestia megaterium sp. strain IMGN3 was isolated from the soil in South Korea. Here, we report its draft genome sequence, comprising 12 contigs with a total sequence length of 5.64 Mbp. This genome will provide valuable resources for future genomic studies, particularly focusing on plant growth promotion and biocontrol.
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In past few years, salinity has become one of the important abiotic stresses in the agricultural fields due to anthropogenic activities. Salinity is leading towards yield losses due to soil infertility and increasing vulnerability of crops to diseases. Fluorescent pseudomonads are a diverse group of soil microorganisms known for promoting plant growth by involving various traits including protecting crops from infection by the phytopathogens. In this investigation, salt tolerant plant growth promoting bacterium Pseudomonas hunanensis SPT26 was selected as an antagonist against Fusarium oxysporum, causal organism of fusarium wilt in tomato. P. hunanensis SPT26 was found capable to produce various antifungal metabolites. Characterization of purified metabolites using Fourier transform infrared spectroscopy (FT-IR) and liquid chromatography-electron spray ionization-mass spectrometry (LC-ESI/MS) showed the production of various antifungal compounds viz., pyrolnitrin, pyochelin and hyroxyphenazine by P. hunanensis SPT26. In the preliminary examination, biocontrol activity of purified antifungal metabolites was checked by dual culture method and results showed 68%, 52% and 65% growth inhibition by pyrolnitrin, 1- hydroxyphenazine and the bacterium (P. hunanensis SPT26) respectively. Images from scanning electron microscopy (SEM) revealed the damage to the mycelia of fungal phytopathogen due to production of antifungal compounds secreted by P. hunanensis SPT26. Application of bioinoculant of P. hunanensis SPT26 and purified metabolites significantly decreased the disease incidence in tomato and increased the plant growth parameters (root and shoot length, antioxidant activity, number of fruits per plant, etc.) under saline conditions. The study reports a novel bioinoculant formulation with the ability to promote plant growth parameters in tomato in presence of phytopathogens even under saline conditions.
Subject(s)
Antifungal Agents , Fusarium , Plant Diseases , Pseudomonas , Solanum lycopersicum , Fusarium/drug effects , Fusarium/growth & development , Fusarium/metabolism , Solanum lycopersicum/microbiology , Solanum lycopersicum/growth & development , Plant Diseases/microbiology , Plant Diseases/prevention & control , Pseudomonas/metabolism , Antifungal Agents/pharmacology , Antifungal Agents/metabolism , Salinity , Biological Control Agents/metabolism , Biological Control Agents/pharmacology , Spectroscopy, Fourier Transform Infrared , Soil Microbiology , Plant Roots/microbiologyABSTRACT
Escherichia coli is one of the most prevalent foodborne pathogens, frequently found in meat and dairy products. Current decontamination methods are often associated with changes in organoleptic characteristics, nutrient loss, and potentially harmful side effects. Furthermore, despite the array of available methods, foodborne outbreaks still frequently occur. For this reason, bacteriophages (or simply phages) emerged as a natural alternative for the biocontrol of bacterial contamination in food without altering their organoleptic properties. In this study, the potential of phage phT4A was assessed in the biocontrol of E. coli in liquid (milk) and solid (ham) food matrices. Firstly, as foods have different pH and temperature values, the influence of these parameters on phage phT4A viability was also assessed to develop an effective protocol. Phage phT4A proved to be stable for long storage periods at pH 7-8 (56 days) and temperatures of 4-37 °C (21 days). Before application of phages to inactivate pathogenic bacteria in food, previous assays were carried out in Tryptic Soy Broth (TSB) to study the dynamics of phage-bacteria interaction. Then, the antibacterial potential of phage phT4A was evaluated in the two food matrices at different temperatures (4, 10 and 25 °C). This phage was more efficient at 25 °C in all tested matrices (maximum inactivation of 6.6, 3.9 and 1.8 log CFU/mL in TSB, milk and ham, respectively) than at 10 °C (maximum decrease of 4.7, 2.1 and 1.0 log CFU/mL in TSB, milk and ham, respectively) and 4 °C (maximum reduction of 2.6 and 0.7 log CFU/mL in TSB and milk, respectively). However, the decrease of temperature from 25 °C to 10 and 4 °C prevented bacterial regrowth. The results suggest that during phage treatment, a balance between an incubation temperature that provide effective results in terms of bacterial inactivation by the phages and at the same time prevents or delays bacterial regrowth, is needed. The application of phage phT4A at a temperature of 10 °C can be an effective strategy in terms of bacterial inactivation, delaying bacterial regrowth and also reducing energy costs.
ABSTRACT
BACKGROUND: Soil-borne plant diseases represent a severe problem that negatively impacts the production of food crops. Actinobacteria play a vital role in biocontrolling soil-borne fungi. AIM AND OBJECTIVES: The target of the present study is to test the antagonistic activity of chitinase-producing Streptomyces cellulosae Actino 48 (accession number, MT573878) against Rhizoctonia solani. Subsequently, maximization of Actino 48 production using different fermentation processes in a stirred tank bioreactor. Finally, preparation of bio-friendly formulations prepared from the culture broth of Actino 48 using talc powder (TP) and bentonite in a natural as well as nano forms as carriers. Meanwhile, investigating their activities in reducing the damping-off and root rot diseases of peanut plants, infected by R. solani under greenhouse conditions. RESULTS: Actino 48 was found to be the most significant antagonistic isolate strain at p ≤ 0.05 and showed the highest inhibition percentage of fungal mycelium growth, which reached 97%. The results of scanning electron microscope (SEM) images analysis showed a large reduction in R. solani mycelia mass. Additionally, many aberrations changes and fungal hypha damages were found. Batch fermentation No. 2, which was performed using agitation speed of 200 rpm, achieved high chitinase activity of 0.1163 U mL- 1 min- 1 with a yield coefficient of 0.004 U mL- 1 min- 1 chitinase activity/g chitin. Nano-talc formulation of Actino 48 had more a significant effect compared to the other formulations in reducing percentages of damping-off and root rot diseases that equal to 19.05% and 4.76% with reduction percentages of 60% and 80%, respectively. The healthy survival percentage of peanut plants recorded 76.19%. Furthermore, the nano-talc formulation of Actino 48 was sufficient in increasing the dry weight of the peanut plants shoot, root systems, and the total number of peanut pods with increasing percentages of 47.62%, 55.62%, and 38.07%, respectively. CONCLUSION: The bio-friendly formulations of actinobacteria resulting from this investigation may play an active role in managing soil-borne diseases.
Subject(s)
Arachis , Chitinases , Fermentation , Plant Diseases , Rhizoctonia , Streptomyces , Streptomyces/enzymology , Rhizoctonia/physiology , Chitinases/metabolism , Arachis/microbiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Roots/microbiologyABSTRACT
Plant parasitic nematodes (PPNs) pose a significant threat to global crop productivity, causing an estimated annual loss of US $157 billion in the agriculture industry. While synthetic chemical nematicides can effectively control PPNs, their overuse has detrimental effects on human health and the environment. Biocontrol agents (BCAs), such as bacteria and fungi in the rhizosphere, are safe and promising alternatives for PPNs control. These BCAs interact with plant roots and produce extracellular enzymes, secondary metabolites, toxins, and volatile organic compounds (VOCs) to suppress nematodes. Plant root exudates also play a crucial role in attracting beneficial microbes toward infested roots. The complex interaction between plants and microbes in the rhizosphere against PPNs is mostly untapped which opens new avenues for discovering novel nematicides through multi-omics techniques. Advanced omics approaches, including metagenomics, transcriptomics, proteomics, and metabolomics, have led to the discovery of nematicidal compounds. This review summarizes the status of bacterial and fungal biocontrol strategies and their mechanisms for PPNs control. The importance of omics-based approaches for the exploration of novel nematicides and future directions in the biocontrol of PPNs are also addressed. The review highlighted the potential significance of multi-omics techniques in biocontrol of PPNs to ensure sustainable agriculture.
ABSTRACT
The seed serves as the primary source for establishing microbial populations in plants across subsequent generations, influencing plant growth and overall health. Cropping conditions, especially farming practices, can influence the composition and functionality of the seed microbiome. Very little is known about the differences in seed microbiome between organic and conventional production systems. In this study, we characterized the endophytic microbial populations in seeds of rice grown under organic and conventional management practices through culture-dependent and -independent analyses. The V4 region of 16S rRNA was used for bacterial taxa identification, and the ITS1 region was used for the identification of fungal taxa. Our results revealed significantly higher Shannon and Simpson indices for bacterial diversity in the conventional farming system, whereas the fungal diversity was higher for observed, Shannon, and Simpson indices in the organic farming system. The cultivable endophytic bacteria were isolated and identified using the full-length 16S rRNA gene. There was no difference in culturable endophytic bacterial isolates in rice seeds grown under both conventional and organic farming systems. Among 33 unique isolates tested in vitro, three bacteria-Bacillus sp. ST24, Burkholderia sp. OR5, and Pantoea sp. ST25-showed antagonistic activities against Marasmius graminum, Rhizoctonia solani AG4, and R. solani AG11, the fungal pathogens causing seedling blight in rice. IMPORTANCE: In this paper, we studied the differences in the endophytic microbial composition of rice seeds grown in conventional and organic farming systems. Our results demonstrate a greater bacterial diversity in conventional farming, while organic farming showcases a higher fungal diversity. Additionally, our research reveals the ability of seed bacterial endophytes to inhibit the growth of three fungal pathogens responsible for causing seedling blight in rice. This study provides valuable insights into the potential use of beneficial seed microbial endophytes for developing a novel microbiome-based strategy in the management of rice diseases. Such an approach has the potential to enhance overall plant health and improve crop productivity.
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The Bacillus velezensis strain NT35, which has strong biocontrol ability, was isolated from the rhizosphere soil of Panax ginseng. The antifungal effects of the NT35 strain against the mycelium and spore growth of Ilyonectria robusta, which causes ginseng rusty root rot, were determined. The inhibitory rate of I. robusta mycelial growth was 94.12% when the concentration of the NT35 strain was 107 CFU·mL-1, and the inhibitory rates of I. robusta sporulation and spore germination reached 100 and 90.31%, respectively, when the concentration of the NT35 strain was 104 and 108 CFU·mL-1, respectively. Strain NT35 had good prevention effects against ginseng rust rot indoors and in the field with the control effect 51.99%, which was similar to that of commercial chemical and biocontrol agents. The labeled strain NT35-Rif160-Stre400 was obtained and colonized ginseng roots, leaves, stems and rhizosphere soil after 90 days. Bacillus velezensis NT35 can induce a significant increase in the expression of five defensive enzyme-encoding genes and ginsenoside biosynthesis-related genes in ginseng. In the rhizosphere soil, the four soil enzymes and the microbial community improved during different periods of ginseng growth in response to the biocontrol strain NT35. The NT35 strain can recruit several beneficial bacteria, such as Luteimonas, Nocardioides, Sphingomonas, and Gemmatimonas, from the rhizosphere soil and reduce the relative abundance of Ilyonectria, Fusarium, Neonectria and Dactylonectria, which cause root rot and rusty root rot in ginseng plants. The disease indices were significantly negatively correlated with the abundances of Sphingomonas and Trichoderma. Additionally, Sphingomonadales, Sphingomonadaceae and actinomycetes were significantly enriched under the NT35 treatment according to LEfSe analysis. These results lay the foundation for the development of a biological agent based on strain NT35.
ABSTRACT
BACKGROUND: Worldwide, vegan and vegetarian lifestyles, as well as food allergies and intolerance (e.g. lactose intolerance and milk protein allergy) demand the development of alternatives to dairy-based probiotic foods. In the present study, probiotic Lacticaseibacillus casei CECT 9104 was added to alginate-based edible coatings enriched with inulin and oligofructose and applied to fresh-cut apple. Microbiological, physicochemical and sensory quality parameters of the apple cubes were monitored during 8 days of refrigerated storage. Lacticaseibacillus casei was tested for its antagonistic effect against inoculated Listeria innocua and Escherichia coli O157:H7. The viability of the probiotic strain during refrigerated storage and after simulated gastrointestinal digestion (GID) was evaluated. RESULTS: After 8 days of storage, 9.52-9.64 log colony-forming units (CFU) g-1 of L. casei were detected in apple samples. The functional apple cubes retained 8.31-8.43 log CFU g-1 of the probiotic after GID, without a significant effect of prebiotic addition. The microbiological quality and nutritional properties were maintained by the use of active coatings, whereas the sensory quality decreased after 8 days of storage. A bactericidal effect was exerted by the probiotic strain loaded in the coating against L. innocua artificially inoculated on apple cubes. Escherichia coli O157:H7 counts were reduced by 2.5 log after 8 days. CONCLUSION: This study has demonstrated the suitability of apple cubes as an alternative matrix to milk for carrying probiotic L. casei CECT 9104 and prebiotics, offering a promising alternative for the development of plant-based functional foods. © 2024 Society of Chemical Industry.