Search Results (615)

With the large-scale cultivation of transgenic Bacillus thuringiensis (Bt) crops, the Bt toxin released from Bt crops is continuously introduced into the soil. Its environmental fate represents a key indicator for assessing the ecological safety of transgenic crops. However, the persistence of Bt toxin in soil is influenced by both biotic and abiotic processes, and their respective contributions under natural conditions remain unclear. This study measured water-dissolved Bt toxin concentrations in paddy soil (PS) and red soil (RS) to compare the influence of biotic and abiotic factors on the dynamic retention of exogenous Bt toxin under different sterilization methods: no sterilization, heat sterilization (HT), and irradiation sterilization (IS). The water-dissolved Bt toxin exhibited a dynamic decrease–increase–decrease trend across all three treatments in both soil types during the 30 day experimental period. Bt toxin displayed rapid adsorption during the initial 2 h stage in RS, but subsequently showed a high desorption, whereas PS probably achieved more stable bonding through soil organic matter (SOM). Different sterilization methods significantly influenced the results by altering abiotic factors: Compared to CK, HT affected soil physicochemical properties and enhanced adsorption resilience, whereas IS caused minimal impact on the soil physicochemical properties, thereby providing a more accurate reflection of abiotic processes. And microbial, as biotic facters, also influence the reduction process of Bt toxin by participating in the adsorption–desorption–degradation equilibrium process. Therefore, we infer that over time, the concentration of water-soluble Bt proteins in the soil will tend toward zero. Additionally, the initial Bt toxin concentration influenced dynamic balance by adjusting adsorption site saturability, with more pronounced desorption reversibility at 500 ng/g concentrations. Overall, this study systematically reveals the effects of soil properties, microorganisms, and sterilization methods on Bt toxin persistence. The findings underscore the importance of selecting and justifying sterilization methods in related environmental behavior studies, while providing essential guidance for the scientific assessment of environmental risks posed by transgenic crops. Full article

The artificially modified Bacillus thuringiensis (Bt) protein can target lepidopteran pests, and planting genetically modified crops with insect-resistant traits is environmentally friendly. However, it is still uncertain whether the exogenous insect-resistant proteins in genetically modified crops will affect the soil rhizosphere microorganisms. This study utilized 16S rDNA sequencing technology to analyze the rhizosphere soil of insect-resistant genetically modified corn LD05 and its control variety Zheng58 at five developmental stages: before sowing, seedling stage, jointing stage, silk emergence stage, and maturity stage. Each sample was taken with six biological replicates, resulting in a total of 60 sequencing samples, with an average of 4368 OTUs obtained per sample. Both alpha and beta analyses showed that LD05 and Zheng58 did not have a significant impact on the soil rhizosphere microbial community. The developmental stage rather than the variety was the main factor causing differences in the bacterial community. Overall, there was no significant difference in the bacterial diversity between the insect-resistant genetically modified corn LD05 and its control variety Zheng58. The results provide useful information for understanding the impact of genetically modified crops on soil microbial communities and also provide a theoretical basis for the safety evaluation of LD05. Full article

Saline–alkali stress is a severe abiotic factor that limits plant growth and development. Endophytic bacteria can improve plant tolerance to such stress through various mechanism, including osmoregulatory substance accumulation and antioxidant enzyme activity. In this study, four saline–alkali-tolerant endophytic strains, designated SYM-2, SYM-4, SYM-9, and SYM-15, were isolated from the roots of alfalfa grown in saline–alkali soil. Though 16S rDNA sequencing, morphological observations, and physiological–biochemical characterization, the strains were identified as closely related to Bacillus cereus, B. thuringiensis, B. halotolerans, and Pantoea agglomerans, respectively. These strains demonstrated the ability to produce 1-aminocyclopropane-1-carboxylate (ACC) deaminase, siderophores, and indole-3-acetic acid (IAA), as well as solubilizing phosphorus. Under saline–alkali conditions, inoculation with these strains significantly increased alfalfa growth parameters. Plant height increased by 4.07–33.90% and root length by 7.49–27.94%, and fresh and dry weight (both above and below ground) increased compared with the control. Strain SYM-15 showed the highest promoting effects, increasing plant height by 33.90%, root length by 27.94% and shoot dry weight by 59.26%. Additionally, root activity increased by 11.23–40%, proline content by 19.09–129–87%, and soluble protein by 7.71–42.49%, and the activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) were significantly elevated across treatments. At the same time, inoculation reduced the levels of hydrogen peroxide (H₂O₂), superoxide anion (O₂⁻), and malondialdehyde (MDA). Compared with the control and other treatments, including SYM-9, the peroxidase activity and superoxide dismutase activity of alfalfa significantly increased after the SYM-15 treatment, while hydrogen peroxide content, phosphorus content, and neutral detergent fiber and acid detergent fiber contents decreased (p < 0.05). Therefore, SYM-15 plays an important role in promoting growth and represents a promising, high-quality strain resource for the large-scale development of microbials aimed at improving alfalfa tolerance under saline–alkali conditions. Full article

Functional microbial inoculation is widely applied in soil restoration; however, its effects on aggregate-scale nutrient cycling remain unclear. Based on ecological stoichiometry theory, we conducted 1-year and 3-year pot experiments using Bacillus thuringiensis (NL-11) and Gongronella butleri (NL-15) under plant-present and plant-absent conditions, with only NL-11 applied in the 1-year experiment. Aggregate size distribution, mean weight diameter (MWD), soil nutrients, microbial biomass, and enzyme activities were evaluated across aggregate classes. The results demonstrated that microbial effects were dependent on both time and plant presence. Under 3-year plant-present conditions, NL-11 and NL-15 significantly increased macroaggregate proportions and MWD, thereby enhancing aggregate stability. Under 3-year no-plant conditions, NL-15 increased organic carbon and total nitrogen in macro- and meso-aggregates by 55–59% and elevated soil C/P and N/P ratios, whereas NL-11 primarily enhanced total nitrogen. In 1-year no-plant macroaggregates, NL-11 increased microbial biomass phosphorus and reduced microbial biomass C/P and N/P ratios. Both inoculants enhanced invertase activity under plant-absent conditions, whereas plant presence stimulated acid phosphatase activity, with NAG activity increasing only under NL-15. Overall, microbial inoculation altered nutrient availability and microbial metabolic characteristics, promoted coordinated C–N–P stoichiometry, and facilitated the recovery of aggregate-scale nutrient cycling. Full article

Empoasca onukii severely damages tea plants as a major sap-sucking pest, leading to the increasing adoption of biopesticides as a sustainable alternative to chemical control. However, existing research has largely focused on the final lethal effects of these agents, while their short-term interference patterns on pest feeding behavior remain unclear. In this study, six biopesticides—azadirachtin, matrine, Beauveria bassiana, Metarhizium anisopliae CQMa421, Mamestra brassicae nucleopolyhedrovirus (MbNPV), and Bacillus thuringiensis (Bt)—were evaluated using the electrical penetration graph (EPG) technique to precisely analyze their interference on the short-term (6 h) feeding behavior of E. onukii, alongside field trials to validate control efficacy. EPG analysis revealed that different types of biopesticides significantly disrupted feeding in distinct ways. The two botanical pesticides and CQMa421 mainly prolonged the non-probing phase (waveform Np) and reduced active non-phloem feeding (C waveform) (p < 0.05); Bt and B. bassiana significantly extended the resting phase (waveform R) and decreased the frequency of passive phloem feeding (waveform E) (p < 0.05), whereas MbNPV exhibited a combined effect, simultaneously prolonging both Np and R waveforms while reducing waveform C (p < 0.05). Field trials showed that all tested treatments achieved complete control (100%) at 21 days post-application. Moreover, across a wide range of concentrations, they all demonstrated excellent and stable control performance. These findings provide diverse agent options for the green control of E. onukii in tea plantations and lay a foundation for constructing a green integrated pest management system centered on biological control for tea plant pests. Full article

Exogenous protein degradation dynamics during transgenic maize straw degradation in soil and the mechanisms underlying soil microbial community construction remain unclear. Applying null-model analysis to determine these mechanisms is important for assessing transgenic crop straw return-to-field-related impacts on dynamic soil quality and microbial ecological function changes. A laboratory leaf degradation burial simulation was conducted to establish an exogenous protein Cry1A.401 soil degradation model and clarify its behaviors. Coupled Illumina MiSeq 16S rDNA sequencing–soil physicochemical factor analysis was used to evaluate soil microbial community characteristic and diversity changes during leaf degradation and explore soil microbial community construction mechanisms and driving factors. The results revealed that exogenous protein Cry1A.401 released from transgenic insect-resistant maize leaves exhibited consistent degradation characteristics, decreasing rapidly at the initial stage but slowly at the middle/late stages. The diversity levels within/between soil microbial community groups did not significantly differ. Coexistence was the dominant interaction type among soil microbial communities. Community assembly occurred stochastically and was limited primarily by diffusion. Insights into the putative mechanistic links among Bacillus thuringiensis (Bt) proteins, soil properties, and microorganisms are provided. Our understanding of the ecological impacts of exogenous Bt proteins released into soil via leaves on soil ecosystems was enhanced. Full article

Bacillus thuringiensis is the most extensively studied entomopathogenic bacterium worldwide; however, its sublethal effects on beetles remain poorly characterized. The aim of this study was to evaluate the toxicity of a previously selected Argentine strain of B. thuringiensis on second-instar Alphitobius diaperinus larvae during an initial 14 days of exposure, and to assess its effects at day 14 and throughout the remainder of the life cycle until death. Three treatments were applied: control, LC₃₀, and LC₅₀. Larval, pupal, and adult weight and body surface area were recorded, and nutritional composition was quantified using colorimetric methods. Insect status was monitored every 48–72 h over a total period of 540 days, until the death of the last individual. Among the evaluated variables, statistically significant differences between control and treatment groups were detected in larval area and weight, in the survival analysis and in two nutritional components: total protein and lipid content per larva. Overall, the results demonstrate that initial sublethal exposure to B. thuringiensis induces chronic lethal effects with delayed mortality in A. diaperinus, indicating irreversible physiological damage. This provides valuable information not only for understanding the biology of this insect but also for stakeholders involved in the productive scaling of beetle-targeted bioinputs. Full article

The Bacillus cereus group (B. cereus group) comprises several closely related species that share high genetic similarity but display markedly different phenotypic traits and pathogenic potential. Reliable and rapid discrimination at the species level remains challenging using conventional microbiological and molecular methods. In this study, Fourier Transform Infrared (FTIR) spectroscopy was evaluated as a rapid phenotypic approach to differentiate seven members of the Bacillus cereus sensu stricto (B. cereus s.s.), Bacillus anthracis (B. anthracis), Bacillus thuringiensis (B. thuringensis), Bacillus mycoides (B. mycoides), Bacillus toyonensis (B. toyonensis), Bacillus wiedmannii (B. wiedmannii) and Bacillus weihenstephanensis (B. weihenstephanensis). A collection of 190 isolates previously characterized by whole genome sequencing was analyzed using the IR Biotyper system. Spectral data were processed through multivariate analyses, including principal component analysis and linear discriminant analysis, following a hierarchical classification strategy. FTIR spectroscopy enabled clear discrimination of B. anthracis from other members of the B. cereus group and allowed the separation of several additional species based on distinct spectral signatures. A further discrimination step permitted differentiation between B. cereus sensu stricto and B. thuringiensis, with minimal overlap. These findings demonstrate that FTIR spectroscopy represents a promising and rapid tool for species-level discrimination within the B. cereus group. While the results should be considered preliminary for species represented by a limited number of isolates, this approach shows strong potential as a complementary method to molecular techniques in routine diagnostics in food safety and veterinary microbiology. Full article

Insecticidal proteins derived from Bacillus thuringiensis (Bt) have been effectively employed in controlling lepidopteran pests, notably in transgenic crops targeting Spodoptera species. However, concerns have arisen regarding the long-term efficacy due to the emergence of tolerant and resistant insect populations. Prior research suggested that repeated exposures to Bt, which contains a mixture of spores and crystals, may contribute to the development of tolerance; however, the specific effects of sequential exposure to purified Cry1 and Vip3Aa proteins remain unclear. This study aimed to assess whether prior exposure of Spodoptera exigua neonate larvae to sublethal concentrations of Cry1Ab, Cry1Ca or Vip3Aa proteins would heighten their tolerance upon subsequent exposure, and whether such effects would extend to their offspring. Pre-exposure to Cry1Ab or Vip3Aa did not affect larval responses to the toxin. For Cry1Ca, a slight increase was observed under one treatment condition, but the effect was not considered biologically relevant. Transgenerational analysis revealed no enhancement of tolerance; rather, there was a negative impact on the offspring’s response in some cases. These findings indicate that although previous studies have documented that sublethal contact with bacterial preparations may significantly affect insect tolerance, exposure to sublethal doses of purified Cry1 and Vip3Aa proteins is unlikely to lead to the development of tolerance in S. exigua. Full article

Bacillus species are among the most widely used microbial agents in agricultural biocontrol, reflecting their ecological resilience, functional diversity, and long history of practical application. The antagonistic activity of Bacillus spp. against plant pathogens and their plant growth–promoting effects are well established. However, these biological functions are frequently considered in isolation from safety evaluations and regulatory decision-making, resulting in a fragmented evidence base. This review addresses this gap by providing an integrated synthesis of agriculturally relevant Bacillus taxa, explicitly linking biocontrol performance with strain-level safety considerations and regulatory assessment. This review focuses on the principal groups currently applied in agriculture, including the Bacillus subtilis lineage, notably B. amyloliquefaciens, B. velezensis, B. pumilus, and B. licheniformis, as well as B. thuringiensis and Cytobacillus firmus. Key mechanisms underlying biocontrol efficacy are examined alongside evidence from greenhouse and field applications. These mechanisms include the production of secondary metabolites and volatile compounds, biofilm formation, rhizosphere colonisation, and the induction of plant defence responses. Attention is given to environmental and operational factors that influence the consistency of performance. A central contribution of this review is the integration of functional evidence with safety-relevant considerations, such as realistic metabolite exposure, antimicrobial resistance potential, and ecological effects. Regulatory approaches in the European Union, the United States, and selected Organisation for Economic Co-operation and Development countries are compared to illustrate how such evidence informs risk assessment and supports the sustainable use of Bacillus-based biocontrol agents in modern agriculture. Full article

The Santiago River near the Guadalajara Metropolitan Area is one of the most contaminated water bodies in Mexico, where heavy metals pose a major threat to aquatic ecosystems. Chronic metal pollution has promoted the adaptation of native microbial communities, including the production of metal-chelating metabolites such as siderophores, which represent a valuable resource for remediation-oriented biomaterials. In this study, bacterial strains were isolated from water and sediment samples, then screened for siderophore production using the Chrome Azurol S assay (CAS), complemented by a MATLAB-based image processing approach for semi-quantitative ranking prior to taxonomic identification by MALDI-TOF MS. Based on biosafety considerations and cultivation robustness, Bacillus thuringiensis was selected as a benchmark case, being immobilized onto activated carbon to produce a carbon–bacteria biocomposite (CBM). To evaluate the performance of CBM, Cu(II) was used as a model contaminant due to its industrial relevance, persistence, toxicity, and strong complexation behavior. Batch adsorption experiments showed that the CBM exhibited a 23.9% higher maximum Cu(II) sorption capacity than pristine activated carbon. Acute toxicity assays using Vibrio fischeri further indicated reduced toxicity in CBM-treated effluents, supporting the feasibility of this contained biocomposite for heavy metal remediation. Full article

Bacillus thuringiensis (Bt) is one of the most extensively used microbial insecticides, attributed to the action of insecticidal crystal proteins (ICPs), primarily Cry toxins, which mediate damage to the insect midgut epithelium. Recent evidence suggests that Bt toxicity is also strongly influenced by its physiological state and interactions with the host gut environment. Biofilm formation represents an important adaptive strategy that enhances bacterial stress tolerance and may modulate insecticidal performance, although the underlying mechanisms remain unclear. However, it is still unclear how Bt in the biofilm state alters host responses at the structural and transcriptomic levels. Using the tea plantation pest Ectropis grisescens as a model, we systematically evaluated the insecticidal efficacy of biofilm-state Bt formulations and their synergistic effects with a biofilm inducer system composed of Tween-80, tea saponin, matrine, and tea polyphenols. Bioassays showed that the biofilm-state Bt supplemented with composite inducers achieved the highest corrected mortality and reduced the LC₅₀ against neonate larvae by 2.88-fold compared with conventional planktonic Bt. Histopathological, biochemical, and transcriptomic analyses further revealed that biofilm-state Bt caused more severe midgut damage and induced extensive remodeling of detoxification- and stress-response-related pathways. These findings highlight Bt physiological state as a critical determinant of formulation efficacy and provide a novel framework for Bt optimization through microbial physiological regulation. Full article

Field-evolved resistance of Helicoverpa zea to crops expressing Cry insecticidal proteins from Bacillus thuringiensis (Bt) is widespread across the United States. To comparatively evaluate physiological factors associated with Bt susceptibility, we analyzed two laboratory strains (Benzon and SIMRU) and one field colony obtained from a commercial corn field near Pickens, Arkansas. Biochemical assays of larval midgut extracts showed that Pickens exhibited significantly altered activities of chymotrypsin-like proteases, aminopeptidase N (APN), and alkaline phosphatase (ALP) compared with the SIMRU or Benzon colonies, with differences varying by larval instar. In contrast, trypsin-like protease activities did not differ significantly among the three colonies. Gene expression analyses of ten serine protease genes and seven candidate Cry receptor genes (including cadherin, ATP-binding cassette family C2, ALP, and four APN genes) revealed significant transcriptional differences in the Pickens relative to the lab colonies. Collectively, these results suggest that chymotrypsin-like proteases may play an important role in the activation of Cry toxins in H. zea. Altered chymotrypsin and APN activities, together with differential gene expressions in the Pickens population, likely contribute to reduced Bt susceptibility. The biochemical and molecular differences provide insight into potential physiological factors underlying reduced Bt susceptibility and may inform future Bt resistance monitoring and management strategies. Full article

Conventional cancer therapies often harm healthy tissues due to their poor specificity, resulting in significant side effects that diminish patients’ quality of life. Parasporins, a group of non-hemolytic parasporal proteins produced by Bacillus thuringiensis, are known for their selective cytotoxicity toward cancer cells. Typically, these proteins require activation through physical or biochemical treatments that fragment them into multiple peptides of varying sizes, which are then tested as mixtures, without purification, against cancer cell lines. In this study, a purification strategy that isolates the protein without prior activation and evaluates the resulting cytotoxic mechanism is proposed. The purification consists of four steps: (1) crystal solubilization with Laemmli buffer, (2) size-based separation via SDS-PAGE, (3) electroelution of the target protein from the gel, and (4) dialysis to remove the elution buffer. From the B. thuringiensis AX isolate, four proteins ranging from ~20 to 60 kDa were recovered, but only AX-2 displayed cytotoxic activity toward MCF-7 breast cancer cells, while remaining non-hemolytic and non-toxic to normal cells (erythrocytes, PBMCs, and MRC-5 fibroblasts). Thus, AX-2 qualifies as a parasporin. AX-2 induces apoptosis in MCF-7 breast cancer cells without generating oxidative stress, and the observed cell death appears to initiate at the plasma membrane rather than through intracellular pathways. Full article

Bt Cry toxins remain the cornerstone of transgenic crop protection against Lepidopteran pests, yet field-evolved resistance, particularly in invasive species such as Spodoptera frugiperda and Helicoverpa armigera, can threaten their long-term efficacy. This review presents a comprehensive and unified mechanistic framework that synthesizes current understanding of Bt Cry toxin modes of action and the complex, multilayered regulatory mechanisms of field-evolved resistance. Beyond the classical pore-formation model, emerging evidence highlights signal transduction cascades, immune evasion via suppression of Toll/IMD pathways, and tripartite toxin–host–microbiota interactions that can dynamically modulate protoxin activation and receptor accessibility. Resistance arises from target-site alterations (e.g., ABCC2/ABCC3, Cadherin mutations), altered midgut protease profiles, enhanced immune regeneration, and microbiota-mediated detoxification, orchestrated by transcription factor networks (GATA, FoxA, FTZ-F1), constitutive MAPK hyperactivation (especially MAP4K4-driven cascades), along with preliminary emerging findings on non-coding RNA involvement. Countermeasures now integrate synergistic Cry/Vip pyramiding, CRISPR/Cas9-validated receptor knockouts revealing functional redundancy, Domain III chimerization (e.g., Cry1A.105), phage-assisted continuous evolution (PACE), and the emerging application of AlphaFold3 for structure-guided rational redesign of resistance-breaking variants. Future sustainability hinges on system-level integration of single-cell transcriptomics, midgut-specific CRISPR screens, microbiome engineering, and AI-accelerated protein design to preempt resistance trajectories and secure Bt biotechnology within integrated resistance and pest management frameworks. Full article