Paenibacillus polymyxa J2-4 induces cucumber to enrich rhizospheric Pseudomonas and contributes to Meloidogyne incognita management under field conditions.

BACKGROUND: Root knot nematodes (RKNs) pose a great threat to agricultural production worldwide. The bacterial nematocides have received increasing attention due to their safe and efficient control against RKNs. Here, we investigated the biocontrol efficacy of Paenibacillus polymyxa J2-4 against Meloidogyne incognita in the field and analyzed the rhizosphere microbiome of cucumber under nematode infection after application of the J2-4 strain. Furthermore, a biomarker strain of Pseudomonas spp. was isolated from the J2-4-inoculated rhizosphere soil, and its nematocidal activity and growth-promoting effect on host plants were determined. In addition, chemotaxis assay of P. fluroescens ZJ5 toward root exudates was carried out. RESULTS: The field experiment demonstrated that P. polymyxa J2-4 could effectively suppressed gall formation in cucumber plants, with the galling index reduced by 67.63% in 2022 and 65.50% in 2023, respectively, compared with controls. Meanwhile, plant height and yield were significantly increased in J2-4 treated plants compared with controls. Metagenomic analysis indicated that J2-4 altered the rhizosphere microbial communities. The relative abundance of Pseudomonas spp. was notably enhanced in the J2-4 group, which was consistent with Linear discriminant analysis Effect Size results that Pseudomonas was determined as one of the biomarkers in the J2-4 group. Furthermore, the ZJ5 strain, one of the biomarker Pseudomonas strains, was isolated from the J2-4-inoculated rhizosphere soil and was identified as Pseudomonas fluorescens. In addition, P. fluorescens ZJ5 exhibited high nematicidal activity in vitro and in vivo, with 99.20% of the mortality rate of M. incognita at 24 h and 69.75% of gall index reduction. The biocontrol efficiency of the synthetic community of ZJ5 plus J2-4 was superior to that of any other single bacteria against M. incognita. Additionally, ZJ5 exhibited great chemotaxis ability toward root exudates inoculated with J2-4. CONCLUSION: Paenibacillus polymyxa J2-4 has good potential in the biological control against M. incognita under field conditions. Enrichment of the beneficial bacteria Pseudomonas fluorescens ZJ5 in the J2-4-inoculated rhizosphere soil contributes to M. incognita management. © 2024 Society of Chemical Industry.

Double-Wing Motif Protein is a Novel Biofilm Regulatory Factor of the Plant Disease Biocontrol Agent, Bacillus subtilis.

Transposon mutagenesis screening of Bacillus subtilis YB-1471, a novel rhizosphere biocontrol agent of Fusarium crown rot (FCR) of wheat, resulted in the identification of orf04391, linked to reduced biofilm formation. The gene encodes a protein possessing a putative tertiary structure of a "double-wing" DNA-binding domain. Expression of orf04391 increased during biofilm development in stationary cultures and during rapid growth in shaking cultures. An orf04391 deletion strain showed reduced biofilm production related to lower levels of the extracellular matrix, and the mutant also had reduced sporulation, adhesion, root colonization, and FCR biocontrol efficiency. Transcriptome analysis of YB-1471 and Δorf04391 in stationary culture showed that the loss of orf04391 resulted in altered expression of numerous genes, including sinI, an initiator of biofilm formation. DNA binding was shown with his-tagged Orf04391 binding to the sinIR operon in vivo and in vitro. Orf04391 appears to be a transcriptional regulator of biofilm formation in B. subtilis through the Spo0A-SinI/SinR pathway.

Assessing the efficacy of phyllospheric growth-promoting and antagonistic bacteria for management of black rot disease of cauliflower incited by Xanthomonas campestris pv. campestris.

The study aimed to assess the potential of phyllospheric bacterial strains isolated from cauliflower plants as biocontrol agents against black rot disease caused by Xanthomonas campestris pv. campestris, through both in vitro and in vivo evaluations. A total of 46 bacterial strains were isolated from healthy and infected cauliflower leaves of both resistant and susceptible plants, and evaluated them for various traits, including plant growth-promoting activities and in vitro antagonistic activity against Xanthomonas campestris pv. campestris. Further, a pot experiment was conducted with the susceptible cauliflower genotype (Pusa Sharad) and 10 selected phyllospheric bacterial isolates to assess their biocontrol efficacy against the disease. The results showed that 82.60% of phyllospheric bacterial isolates were positive for phosphate solubilization, 63.04% for ammonia production, 58.69% for HCN production, 36.95% for siderophore production, and 78.26% had the capacity to produce IAA. Out of the 46 isolates, 23 exhibited in vitro antagonistic activity against X. campestris pv. campestris and 10 isolates were selected for a pot experiment under glasshouse conditions based on their good plant growth-promoting activities and antagonistic assay. The results revealed that bacterial isolate CFLB-27 exhibited the highest biocontrol efficiency (65.41%), followed by CFLB-24 (58.30%), CFLB-31 (47.11%), and CFLB-26 (46.03%). These four isolates were identified as Pseudomonas fluorescens CFLB-27, Bacillus velezensis CFLB-24, Bacillus amyloliquefaciens CFLB-31, and Stenotrophomonas rhizophila CFLB-26. This study provides valuable insights into the potential of phyllospheric bacteria as an effective tool for disease management in sustainable agriculture.

A novel biocontrol agent Bacillus velezensis K01 for management of gray mold caused by Botrytis cinerea.

Gray mold is a destructive plant disease caused by a fungal pathogen Botrytis cinerea. The use of plant growth promoting rhizobacteria (PGPR) has proven to be a promising method to control this disease. Bacillus velezensis K01 was isolated from the rhizosphere of planting tomatoes. Strain K01 has a range of roles, including the ability to solubilize phytate phosphorus, stimulate resistant response, and produce indoleacetic acid (IAA), protease, cellulase, and antimicrobial substances. Strain K01 was found to inhibit 12 phytopathogenic fungi and 5 phytopathogenic bacteria. Specially, strain K01 demonstrated a biocontrol efficiency of over 78% against gray mold caused by B. cinerea on the leaves and fruits of tomato and pepper. Additionally, K01 was found to promote the growth of maize seedlings. Further genomic analysis revealed that K01 belongs to B. velezensis, which is consistent with phylogenetic analysis, average nucleotide polymorphism (ANI), and digital DNA-DNA hybridization (dDDH). The genome of strain K01 had a size of 3,927,799 bp and deduced 3866 predicted genes, with an average guanine-cytosine (GC) content of 46.5%. Based on the analyses of genomic secondary metabolites, over 18.4% of the genome was annotated to 12 gene clusters related to antimicrobial metabolite synthesis. Additionally, genome annotation and comparative genomics identified several genes associated with plant growth promotion and environmental adaption. These findings suggest that B. velezensis K01 has the potential to serve as a new biocontrol agent for management of gray mold on tomato and pepper.

Susceptibility of Hyphantria cunea larvae to Beauveria bassiana under Cd Stress: An integrated study of innate immunity and energy metabolism.

Biological control is widely used for integrated pest management. However, there are many abiotic factors that can affect the biocontrol efficiency. In this study, we investigated the susceptibility of Hyphantria cunea larvae to Beauveria bassiana under Cd stress, and the corresponding mechanism was analyzed around innate immunity and energy metabolism. The results showed that mortality of H. cunea larvae treated with Cd and B. bassiana was significantly higher than those treated with B. bassiana alone, and the combined lethal effect exhibited a synergistic effect. Compared with the single fungal treatment group, the total hemocyte count in the combined Cd and fungal treatment group decreased significantly, accompanied by a decrease in phagocytosis, encapsulation, and melanization activity. The expression levels of three phagocytosis-related genes, one encapsulation-promoting gene, and one melanization-regulating gene were significantly lower in the combined treatment group than those in the single fungal treatment group. Furthermore, pathogen recognition ability, signal transduction level, and immune effector expression level were weaker in the combined treatment group than those in the single fungal treatment group. The expression levels of 14 key metabolites and 7 key regulatory genes in glycolysis and tricarboxylic acid cycle pathways were significantly lower in the combined treatment group than those in the single fungal treatment group. Taken together, the weakness of innate immunity and energy metabolism in response to pathogen infection resulted in an increased susceptibility of H. cunea larvae to B. bassiana under Cd pre-exposure. Microbial insecticide is a preferred strategy for pest control in heavy metal-polluted areas. AVAILABILITY OF DATA AND MATERIAL: All the data that support the findings of this study are available in the manuscript.

Evaluation of the Biocontrol Efficiency of Bacillus subtilis Wettable Powder on Pepper Root Rot Caused by Fusarium solani.

The plant-growth-promoting rhizobacteria (PGPR) B. subtilis PTS-394 has been utilized as a biocontrol agent (in a wettable powder form) due to its excellent ability to suppress tomato soil-borne diseases caused by Fusarium oxysporum and Ralstonia solanacearum. In this study, we evaluated the biocontrol efficiency of Bacillus subtilis PTS-394 wettable powder on pepper root rot in pot experiments and field trials. B. subtilis PTS-394 and its lipopeptide crude extract possessed excellent inhibition activity against Fusarium solani, causing pepper root rot; in an antifungal activity test B. subtilis PTS-394 wettable powder exhibited a good ability to promote pepper seed germination and plant height. The experiments in pots and the field indicated that B. subtilis PTS-394 wettable powder had an excellent control effect at 100-fold dilution, and its biocontrol efficacy reached 69.63% and 74.43%, respectively. In this study, the biocontrol properties of B. subtilis PTS-394 wettable powder on pepper root rot were evaluated and its application method was established. It was concluded that B. subtilis PTS-394 wettable powder is a potential biocontrol agent with an excellent efficiency against pepper root rot.

Cd exposure-triggered susceptibility to Bacillus thuringiensis in Lymantria dispar involves in gut microbiota dysbiosis and hemolymph metabolic disorder.

The immunotoxicity induced by heavy metals on herbivorous insects reflect the alterations of the susceptibility to entomopathogenic agents in herbivorous insects exposed to heavy metal. In the present study, the susceptibility of gypsy moth larvae to Bacillus thuringiensis under Cd treatment at low and high dosages was investigated, and the gut microbiome-hemolymph metabolome responses that affected larval disease susceptibility caused by Cd exposure were examined. Our results showed that mortality of gypsy moth larvae caused by B. thuringiensis was significantly higher in larvae pre-exposed to Cd stress, and there was a synergistic effect between Cd pre-exposure and bacterial infection. Exposure to Cd significantly decreased the abundance of several probiotics (e.g., Serratia for the low Cd dosage and Weissella, Aeroonas, and Serratia for the high Cd dosage) and increased the abundances of several pathogenic bacteria (e.g., Stenotrophomonas, Gardnerella, and Cutibacterium for the low Cd dosage and Pluralibacter and Tsukamurella for the high Cd dosage) compared to the controls. Moreover, metabolomics analysis indicated that amino acid biosynthesis and metabolism were significantly perturbed in larval hemolymph under Cd exposure at both the low and high dosages. Correlation analysis demonstrated that several altered metabolites in larval hemolymph were significantly correlated with changes in the gut microbial community. The results demonstrate that prior exposure to Cd increases the susceptibility of gypsy moth larvae to B. thuringiensis in a synergistic fashion due to gut microbiota dysbiosis and hemolymph metabolic disorder, and thus microbial-based biological control may be the best pest control strategy in heavy metal-polluted areas.

Cd exposure-triggered metabolic disruption increases the susceptibility of Lymantria dispar (Lepidoptera: Erebidae) larvae to Mamestra brassicae nuclear polyhedrosis virus: A multi-omics study.

Biological control is an environmentally friendly and effective pest control strategy, but it is often affected by a variety of abiotic factors in the pest control area. Here, the susceptibility of gypsy moth larvae to Mamestra brassicae nuclear polyhedrosis virus (MbNPV) under Cd treatment at the low and high dosages was investigated, and the mechanism of Cd stress affecting virus susceptibility of gypsy moth larvae was analyzed from a metabolic perspective by combining transcriptome and metabolome of the larval fat body. Our results showed that the mortality of MBNPV infection on gypsy moth larvae pre-exposed to Cd was significantly higher than that of larvae without Cd pre-exposure, and the joint effects of Cd exposure and virus infection on larval mortality were demonstrated to be synergistic. Transcriptome analysis revealed that amino acid and carbohydrate transport and metabolism accounted for most of the differently expressed genes in the low Cd and high Cd treatment groups. Consistent with the transcriptome results, metabolome analysis also showed that most metabolites affected by Cd exposure were involved in amino acid and carbohydrate metabolism. Function analysis showed that the contents of several amino acids (e.g. tryptophan and tyrosine) with antioxidant properties were significantly increased in Cd-treated gypsy moth larvae. Taken together, Cd exposure as an environmental factor, promotes the susceptibility of gypsy moth larvae to MbNPV, and metabolic disruption, especially amino acids and carbohydrates-related metabolism, is responsible for the increased susceptibility of gypsy moth larvae to virus under Cd stress.

Influence of arginine on the biocontrol efficiency of Metschnikowia citriensis against Geotrichum citri-aurantii causing sour rot of postharvest citrus fruit.

This study investigated the effect of arginine (Arg) on the antagonistic activity of Metschnikowia citriensis against sour rot caused by Geotrichum citri-aurantii in postharvest citrus, and evaluated the possible mechanism therein. Arg treatment up-regulated the PUL genes expression, and significantly induced the pulcherriminic acid (PA) production of M. citriensis, which related to the capability of iron depletion of M. citriensis. By comparing the biocontrol effects of Arg-treated and untreated yeast cells, it was found that Arg treatment significantly enhanced the biocontrol efficacy of M. citriensis, and 5 mmol L-1 Arg exerted the best effect. Additionally, the biofilm formation ability of M. citriensis was greatly enhanced by Arg, and the higher population density of yeast cells in citrus wounds was also observed in Arg treatment groups stored both at 25 °C and 4 °C. Moreover, Arg was shown to function as a cell protectant to elevate antioxidant enzyme activity [including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX)] and intracellular trehalose content to resist oxidative stress damage, that directly helped to enhance colonization ability of yeasts in fruit wounds. These results suggest the application of Arg is a useful approach to improve the biocontrol performance of M. citriensis.

Septicaemia of subterranean termites Coptotermes curvignathus caused by disturbance of bacteria isolated from termite gut and its foraging pathways.

Microbial pathogens continue to attract a great deal of attention to manage the termite population. Every bacterium has its own mode of action and in fact, the mechanisms used by bacteria to attack termites remain elusive at the moment. Hence, the objective of this study was to evaluate the susceptibility of subterranean termites Coptotermes curvignathus to opportunistic pathogens using culturable aerobic bacteria isolated from the termite gut and its foraging pathways. Bacterial suspensions were prepared in concentrations of 103, 106 and 109 colony-forming units (CFU) ml-1 and introduced to the termites via oral-contact and physical contact treatment. The data show that contact method acted slower and gave lower mortality, compared to the oral-contact method. Coptotermes curvignathus were highly susceptible to Serratia marcescens and Pseudomonas aeruginosa. Serratia marcescens showed the highest mortality percentage of 68% and 54% at bacterial concentration of 109 CFU ml-1 via oral-contact and contact method, respectively. Serratia marcescens was also defined as the bacteria with the highest ability to induce the high mortality of C. curvignathus with the lowest concentration of bacterial suspension at a given time under laboratory condition. The results of this study indicate that P. aeruginosa and S. marcescens in particular may be attractive candidates worth further examination as a possible biocontrol agent against C. curvignathus in the field and to evaluate environmental and ecological risks of the biocontrol.

Study on screening and antagonistic mechanisms of Bacillus amyloliquefaciens 54 against bacterial fruit blotch (BFB) caused by Acidovorax avenae subsp. citrulli.

Bacterial fruit blotch (BFB) was a serious threat to cucurbitaceae crops. It was caused by the gram-negative bacterium Acidovorax avenae subsp. citrulli. Two hundred strains, which have the potential in controlling plant diseases in our laboratory's biocontrol strain library, were employed to this research to screen some antagonistic bacteria, which can efficiently control bacterial fruit blotch disease. Based on the results of antagonistic activity experiments, greenhouse tests and field trials, 5 of the test strains have high abilities to control BFB. One of the 5 bacteria strains has the highest potential to control BFB named 54. The biocontrol efficacy of 54 was up to 60%. To characterize the strain, we used series of methods to evaluate the bacterium, including morphology analysis, physiological biochemical test and biomolecular assay. We found that the bacterium 54 belongs to the species Bacillus amyloliquefaciens. The colonization test results showed that 54 had the highest colonization levels, and the density of the strain on leaves was up 10(5)colony forming units (CFU) per gram of leaf tissue. Our recent results show that B. amyloliquefaciens 54 can promote the plant growth due to raised the contents of available N, P, K and the leaf chlorophyll. The antagonistic bacterium 54 can significantly control the BF B by increasing the expression level of defense-related gene PR1 and the accumulation the hydrogen peroxide in the plant. The results of trail experiment was also verified this efficient results of bacterium. This is also the first report of B. amyloliquefaciens strain that is able to control BFB.

Isolation of Bacillus amyloliquefaciens S20 and its application in control of eggplant bacterial wilt.

Bacterial strain S20 was isolated and identified as Bacillus amyloliquefaciens based on physiological and biochemical characteristics and a 16S rRNA gene sequence analysis. Strain S20 inhibits the growth of Fusarium oxysporum and Ralstonia solanacearum. Some genes associated with the synthesis of some lipopeptides were detected in strain S20 by PCR. Iturins A were identified as the main antagonistic substrates by analysis with electrospray ionization mass spectrometry/collision-induced dissociation (ESI-MS/CID). Four homologues of iturin A (C13-C16) were identified. Pot experiments showed that the application of strain S20 alone could control eggplant wilt with an efficacy of 25.3% during a 40 day experiment. If strain S20 was used with organic fertilizer, the control efficacy against eggplant wilt reached as high as 70.7%. The application of organic fertilizer alone promotes the growth of R. solanacearum, resulting in a higher wilt incidence than that observed in control plants. The application of strain S20 effectively inhibits R. solanacearum in the rhizosphere soil of eggplant. The combined use of strain S20 and organic fertilizer more effectively controlled R. solanacearum in soil than the use of strain S20 alone. The soil count of strain S20 decreased gradually during the course of the experiment after inoculation. Organic fertilizer was beneficial for the survival of the antagonistic bacterial strain S20; a higher level of these bacteria could be maintained. The application of organic fertilizer with strain S20 increased bacterial diversity in rhizosphere soil.