The RNA-binding domain of DCL3 is required for long-distance RNAi signaling.

Small RNA (sRNA)-mediated RNA silencing (also known as RNA interference, or RNAi) is a conserved mechanism in eukaryotes that includes RNA degradation, DNA methylation, heterochromatin formation and protein translation repression. In plants, sRNAs can move either cell-to-cell or systemically, thereby acting as mobile silencing signals to trigger noncell autonomous silencing. However, whether and what proteins are also involved in noncell autonomous silencing have not been elucidated. In this study, we utilized a previously reported inducible RNAi plant, PDSi, which can induce systemic silencing of the endogenous PDS gene, and we demonstrated that DCL3 is involved in systemic PDS silencing through its RNA binding activity. We confirmed that the C-terminus of DCL3, including the predicted RNA-binding domain, is capable of binding short RNAs. Mutations affecting RNA binding, but not processing activity, reduced systemic PDS silencing, indicating that DCL3 binding to RNAs is required for the induction of systemic silencing. Cucumber mosaic virus infection assays showed that the RNA-binding activity of DCL3 is required for antiviral RNAi in systemically noninoculated leaves. Our findings demonstrate that DCL3 acts as a signaling agent involved in noncell autonomous silencing and an antiviral effect in addition to its previously known function in the generation of 24-nucleotide sRNAs. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-023-00124-6.

Erratum: Publisher Correction: The RNA-binding domain of DCL3 is required for long-distance RNAi signaling.

[This corrects the article DOI: 10.1007/s42994-023-00124-6.].

Low-density polyethylene enhances the disturbance of microbiome and antibiotic resistance genes transfer in soil-earthworm system induced by pyraclostrobin.

Non-antibiotic chemicals in farmlands, including microplastics (MPs) and pesticides, have the potential to influence the soil microbiome and the dissemination of antibiotic resistance genes (ARGs). Despite this, there is limited understanding of the combined effects of MPs and pesticides on microbial communities and ARGs transmission in soil ecosystems. In this study, we observed that low-density polyethylene (LDPE) microplastic enhance the accumulation of pyraclostrobin in earthworms, resulting in reduced weight and causing severe oxidative damage. Analysis of 16 S rRNA amplification revealed that exposure to pyraclostrobin and/or LDPE disrupts the microbial community structure at the phylum and genus levels, leading to reduced alpha diversity in both the soil and earthworm gut. Furthermore, co-exposure to LDPE and pyraclostrobin increased the relative abundance of ARGs in the soil and earthworm gut by 2.15 and 1.34 times, respectively, compared to exposure to pyraclostrobin alone. It correlated well with the increasing relative abundance of genera carrying ARGs. Our findings contribute novel insights into the impact of co-exposure to MPs and pesticides on soil and earthworm microbiomes, highlighting their role in promoting the transfer of ARGs. This knowledge is crucial for managing the risk associated with the dissemination of ARGs in soil ecosystems.

Global analysis of seed transcriptomes reveals a novel PLATZ regulator for seed size and weight control in soybean.

Seed size and weight are important factors that influence soybean yield. Combining the weighted gene co-expression network analysis (WGCNA) of 45 soybean accessions and gene dynamic changes in seeds at seven developmental stages, we identified candidate genes that may control the seed size/weight. Among these, a PLATZ-type regulator overlapping with 10 seed weight QTLs was further investigated. This zinc-finger transcriptional regulator, named as GmPLATZ, is required for the promotion of seed size and weight in soybean. The GmPLATZ may exert its functions through direct binding to the promoters and activation of the expression of cyclin genes and GmGA20OX for cell proliferation. Overexpression of the GmGA20OX enhanced seed size/weight in soybean. We further found that the GmPLATZ binds to a 32-bp sequence containing a core palindromic element AATGCGCATT. Spacing of the flanking sequences beyond the core element facilitated GmPLATZ binding. An elite haplotype Hap3 was also identified to have higher promoter activity and correlated with higher gene expression and higher seed weight. Orthologues of the GmPLATZ from rice and Arabidopsis play similar roles in seeds. Our study reveals a novel module of GmPLATZ-GmGA20OX/cyclins in regulating seed size and weight and provides valuable targets for breeding of crops with desirable agronomic traits.

Effect of fertilization on the degradation and enantioselectivity of fipronil in soil.

BACKGROUND: Fertilizers and pesticides are commonly used simultaneously in agriculture. However, the effects of common fertilizers on the dissipation, enantioselectivity, and metabolites of the chiral insecticide fipronil in soil are yet to be reported. RESULT: An enantioselective method for detecting fipronil enantiomers and their metabolites in different soil matrices was developed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The results showed that organic and compound fertilizers significantly decreased the degradation of S- and R-fipronil, whereas phosphate and microbial fertilizers slightly reduced fipronil dissipation. The half-life values for S- and R-fipronil were 43.3 and 28.9 days, 99.0 and 63.0 days, 69.3 and 43.3 days, 46.2 and 30.1 days, and 43.3 and 31.5 days, respectively, in the control and the four fertilizer treatments, respectively. The enantioselectivity of fipronil enantiomers occurred and R-fipronil exhibited preferential degradation with an enantiomeric fraction (EF) of 0.4900-0.6238 in all treatments; but the four tested fertilizers decreased enantioselectivity with EF values changed from 0.4970 to 0.6238 in the control to 0.4900-0.6171 in fertilizer treatments. Two metabolites, fipronil sulfone and sulfide, were produced, and their amounts increased with culture time in all treatments. Fertilization reduced the content of fipronil sulfide and sulfone but hardly reduced the total amount of fipronil and its metabolites. CONCLUSION: Fertilizers affect the environmental behavior of fipronil in the soil. Fertilization alters the soil bacterial community, which may be an important factor. This influence is relatively complicated and should be comprehensively considered in the environmental risk assessment of pesticides. © 2023 Society of Chemical Industry.

Enantioselective effect of the chiral fungicide tebuconazole on the microbiota community and antibiotic resistance genes in the soil and earthworm gut.

Tebuconazole, consisting of two enantiomers, has a high detectable rate in the soil. The residue of tebuconazole in the soil may cause risk to microbiota community. Antibiotic resistance genes (ARGs) are considered as emerging environmental contaminants, and they can be transferred vertically and horizontally between microbiota community in the soil. Until now, the enantioselective effect of tebuconazole on the microbiota community and ARGs in the soil and earthworm gut has remained largely unknown. Tebuconazole enantiomers showed different bioconcentration behaviors in earthworms. The relative abundances of bacteria belonging to Actinobacteriota, Crenarchaeota and Chloroflexi in R-(-)-tebuconazole-treated soil were higher than those in S-(+)-tebuconazole-treated soil at same concentrations. In the earthworm gut, bacteria belonging to Proteobacteria and Bacteroidota exhibited different relative abundances between the S-(+)-tebuconazole and R-(-)-tebuconazole treatments. The numbers and abundances of ARGs in the soil treated with fungicides were higher than those in the control. In earthworm gut, the diversities of ARGs in all treatments were higher than that in the control, and the relative abundances of Aminoglycoside, Chloramphenicol, Multidrug resistance genes and mobile genetic elements (MGEs) in R-(-)-tebuconazole-treated earthworm gut were higher than those in S-(+)-tebuconazole-treated earthworm gut. Most of ARGs showed a significantly positive correlation with MGEs. Based on network analysis, many ARGs may be carried by bacteria belonging to Bacteroidota and Proteobacteria. These results provide valuable information for understanding the enantioselective effect of tebuconazole on the microbiota community and ARGs.

Effect of Cd/Cu on the toxicity and stereoselective environmental behavior of dinotefuran in earthworms Eisenia foetida.

Pesticides and heavy metals commonly coexist in soil. In this study, the influence of Cd and Cu on the toxicity of rac-dinotefuran and the enantioselective behavior of dinotefuran enantiomers in soil-earthworm microcosms were investigated. The acute toxic tests showed that S-dinotefuran has higher toxic than that of R-dinotefuran. The rac-dinotefuran and Cd has an antagonistic effect on earthworms, and the Cu and rac-dinotefuran has a synergistic effect. Earthworms maybe promoted the enantioselective behavior of dinotefuran in soil. Co-exposure to Cd or Cu inhibited the dissipation of dinotefuran enantiomers (S-dinotefuran and R-enantiomers), and slightly reduced the enantioselectivity in soil. The earthworms were found to be preferentially enriched with S-dinotefuran. However, Cd or Cu attenuated the accumulation of dinotefuran enantiomers in earthworms and decreased the enantioselectivity. The effect of Cd and Cu on the environmental behaviors of dinotefuran enantiomers were correlated positively with the dose of Cd/Cu. These results showed that Cd and Cu alter the environmental behaviors and the toxicity of dinotefuran enantiomers in soil-earthworm microcosms. Thus, the influence of coexistent heavy metals on the ecological risk assessment of chiral pesticides should be considered.

Enantioselective effect of chiral fungicide prothioconazole on Fusarium graminearum: Fungicidal activity and DON biosynthesis.

Prothioconazole, a chiral triazole fungicide, is widely used to control Fusarium head blight (FHB) of wheat. Fusarium graminearum (F. graminearum), as the main pathogen of FHB, can produce many secondary metabolites including deoxynivalenol (DON), which threatens the health of humans and animals. However, some fungicides may stimulate F. graminearum to synthesize more DON under certain conditions. Until now, the fungicidal activity and enantioselective effect of prothioconazole enantiomers on DON production, transcriptome and metabolome of F. graminearum were unclear. The fungicidal activity of R-(-)-prothioconazole against F. graminearum was 9.12-17.73 times higher than that of S-(+)-prothioconazole under all conditions. Prothioconazole enantiomers can induce F. graminearum to synthesize more DON under 0.99 water activity (aw) and 30 °C, especially R-(-)-prothioconazole. The expression levels of TRI6, TRI10 and TRI101 under R-(-)-prothioconazole treatment were significantly higher than those under S-(+)-prothioconazole treatment. Most genes in glycolysis, pyruvate metabolism, the target of rapamycin (TOR) signaling transduction pathway and the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) signaling transduction pathway showed higher expression levels under R-(-)-prothioconazole treatment than uner S-(+)-prothioconazole treatment and the control. The peroxisome pathway displayed higher transcriptional activity under S-(+)-prothioconazole treatment compared with R-(-)-prothioconazole and the control. Based on metabolomic data, R-(-)-prothioconazole can significantly influence phenylalanine metabolism, and no significantly enriched pathway was found under S-(+)-prothioconazole treatment. These results are helpful to understand the risk of prothioconazole enantiomers on DON production of F. graminearum and uncover the relevant underlying mechanisms of prothioconazole enantiomers.

Toxicity, bioactivity of triazole fungicide metconazole and its effect on mycotoxin production by Fusarium verticillioides: New perspective from an enantiomeric level.

The chiral triazole fungicide metconazole has four stereoisomers, is a broad-spectrum fungicide and is widely used for controlling Fusarium head blight caused by Fusarium species. In this study, systemic assessments of metconazole stereoisomers were performed, including stereoselective toxicity toward the aquatic organism Daphnia magna, fungicidal activity and effects on fumonisin production by the pathogen Fusarium verticillioides (F. verticillioides) in relation to different conditions. The toxicity of metconazole was enantioselective, and there was a 2.1-2.9-fold difference. The activities of superoxide dismutase (SOD) and catalase (CAT) increased and decreased, respectively, after treatment with metconazole stereoisomers, and the differences were observed among the stereoisomers. Among the four stereoisomers, (1S,5R)-metconazole showed the highest fungicidal activity under all assayed conditions, and the differences ranged from 4.4 to 45.2 times. Moreover, metconazole stereoisomers can stereoselectively affect on fumonisin B1 production by F. verticillioides and abiotic factors, such as water activity and temperature, play an important role. Our study provides new insight into metconazole at the stereoisomeric level, including its toxicity, bioactivity, and effect on Fusarium species producing mycotoxins.

Enantioselective Effect of Flutriafol on Growth, Deoxynivalenol Production, and TRI Gene Transcript Levels in Fusarium graminearum.

In recent years, deoxynivalenol (DON) has frequently been detected in wheat grains and their products. The enantioselective impact of flutriafol on the growth and DON biosynthesis of Fusarium graminearum was investigated in relation to water activity (αw, 0.97 and 0.99) and temperature (20, 25, and 30 °C) on the wheat-based medium. R-(-)-flutriafol exhibited higher bioactivity than S-(+)-flutriafol and Rac-flutriafol under the above conditions. Flutriafol enantiomers reduced or stimulated DON biosynthesis depending on αw. DON levels were negligible after 14 or 7 days of incubation times under 0.97 and 0.99 aw, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) analyses showed that the expression levels of trichothecene biosynthetic (TRI) genes of F. graminearum under 0.97 aw were significantly higher than those under 0.99 aw. In addition, R-(-)-flutriafol can induce more TRI gene expression than S-(+)-flutriafol. Taken together, this study indicated that aw and temperature play important roles in regulating DON biosynthesis in F. graminearum with flutriafol enantiomers.

Generating Novel Male Sterile Tomatoes by Editing Respiratory Burst Oxidase Homolog Genes.

Hybrid breeding of tomatoes (Solanum lycopersicum), an important vegetable crop, is an effective way to improve yield and enhance disease and stress resistance. However, the efficiency of tomato hybridization is hindered by self-fertilization, which can be overcome using male sterile lines. It has been reported that reactive oxygen species (ROS) act as a key regulator for anther development, mediated by RBOH (Respiratory Burst Oxidase Homolog) genes. Here, two tomato anther-expressed genes, LeRBOH (Solyc01g099620) and LeRBOHE (Solyc07g042460), were selected to cultivate novel tomato male sterile strains. By using a CRISPR/Cas9 system with a two-sgRNA module, the lerboh, lerbohe, and lerboh lerbohe mutant lines were generated, among which the lerbohe and lerboh lerbohe mutants displayed complete male sterility but could accept wild-type pollens and produce fruits normally. Further analysis uncovered significantly decreased ROS levels and abnormal programmed cell death in lerboh lerbohe anthers, indicating a key role of ROS metabolism in tomato pollen development. Taken together, our work demonstrates a successful application of gene editing via CRISPR/Cas9 in generating male sterile tomatoes and afforded helpful information for understanding how RBOH genes regulating tomato reproduction process.

Determination of pyrethroid residues in herbal tea using temperature-controlled ionic liquid dispersive liquid-liquid microextraction by high performance liquid chromatography.

A simple and effective method for determining five pyrethroid residues in herbal tea by ultrasound-enhanced temperature-controlled (UETC) ionic liquid dispersive liquid-liquid microextraction (IL-DLLME) coupled with high performance liquid chromatography-diode array detection (HPLC-DAD) was developed. The use of ultrasonication and heating improved the ability of the ionic liquid to extract the analytes. Various parameters that affect the extraction efficiency were investigated and optimized using single factor experiments and response surface design. The optimum conditions of the experiment were 121 µL of [HMIM][PF6] (extraction solvent), 794 µL of acetonitrile (dispersive solvent), a heating temperature of 40°C, a sonication time of 3.6 min and a pH of 2.9. Under optimized conditions, the linearity was in the range of 0.05-5 mg L-1 with correlation coefficients above 0.9993. The limits of detection and quantification were 1.25-1.35 µg L-1 and 5 µg L-1, respectively. The mean recoveries of the five pyrethroids ranged from 74.02% to 109.01%, with RSDs below 9.04%. The proposed method was reliable for the analysis of pyrethroids in Chinese herbal tea.

Dissipation of fluazinam in citrus groves and a risk assessment for its dietary intake.

BACKGROUND: Citrus is one of the most important fruit crops worldwide. Fluazinam is a fungicide that is used to control fungal diseases, and its dissipation and residue in citrus fruits should be studied. RESULTS: A Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) procedure combined with gas chromatography with an electron capture detector (GC-ECD) has been developed. The fortified recoveries ranged from 82.1% to 105.9%, with relative standard deviations (RSDs) of less than 5.7%. Fluazinam dissipated relatively quickly following first-order kinetics, with a half-life of 8.5-9.5 days. The experiments on the terminal residue of fluazinam in citrus were conducted at six locations in China, and the risk quotient (RQ) method was applied to citrus fruits for dietary exposure risk assessment based on the terminal residue test. The RQs of fluazinam at three preharvest intervals (PHIs) (21, 28, and 35 days) were all less than 100%, which is an acceptable level for human consumption. The present study provides a reference for the establishment of maximum residue limit (MRL) for fluazinam in citrus. CONCLUSIONS: The dissipation and residues of fluazinam in citrus were monitored. The half-life of less than 10 days showed that fluazinam could degrade relatively easily in citrus. The risk assessment also indicated the intake safety of fluazinam in citrus. © 2019 Society of Chemical Industry.

A Vortex-Assisted Dispersive Liquid-Liquid Microextraction Followed by UPLC-MS/MS for Simultaneous Determination of Pesticides and Aflatoxins in Herbal Tea.

A method for detecting the organophosphorus pesticides residue and aflatoxins in China herbal tea has been developed by UPLC-MS/MS coupled with vortex-assisted dispersive liquid-liquid microextraction (DLLME). The extraction conditions for vortex-assisted DLLME extraction were optimized using single-factor experiments and response surface design. The optimum conditions for the experiment were the pH 5.1, 347 µL of chloroform (extraction solvent) and 1614 µL of acetonitrile (dispersive solvent). Under the optimum conditions, the targets were good linearity in the range of 0.1 µg/L⁻25 µg/L and the correlation coefficient above 0.9998. The mean recoveries of all analytes were in the ranged from 70.06%⁻115.65% with RSDs below 8.54%. The detection limits were in the range of 0.001 µg/L⁻0.01µg/L. The proposed method is a fast and effective sample preparation with good enrichment and extraction efficiency, which can simultaneously detect pesticides and aflatoxins in China herbal tea.

Effect of Tebuconazole Enantiomers and Environmental Factors on Fumonisin Accumulation and FUM Gene Expression in Fusarium verticillioides.

Fusarium verticillioides is an important corn pathogen that can produce fumonisins (FBs) under certain environmental conditions. In this study, we evaluated the enantioselective impact of tebuconazole enantiomers on the growth and FB production of F. verticillioides on maize-based media at different abiotic factors. The expression of FB biosynthetic genes ( FUM1 and FUM6) was quantified by real-time reverse transcription polymerase chain reaction. The results showed that water activity ( aw), temperature, and types of tebuconazole significantly affected the growth of F. verticillioides. The order of fungicidal activity was (-)-tebuconazole > rac-tebuconazole > (+)-tebuconazole. (-)-tebuconazole exhibited the maximal selective fungicidal activity (242-fold) against F. verticillioides at 0.95 aw and 35 °C. Production of fumonisin B1 (FB1) and fumonisin B2 (FB2) by F. verticillioides was influenced by aw, temperature, types of tebuconazole, and dose. Under most conditions, (-)-tebuconazole showed stronger inhibition for FB1 and FB2 production than (+)-tebuconazole (1.87-2.85-fold reduction in FBs) and rac-tebuconazole. The optimal environmental condition for FB production was at 0.99 aw and 25 °C. Tebuconazole enantiomers differently affected FB biosynthetic gene ( FUM1 and FUM6) expression, but the effects on FB production and gene expression showed no positive correlation. The present study provides a better understanding on ways to minimize FB production in corn treated with fungicides.

Selective effect of myclobutanil enantiomers on fungicidal activity and fumonisin production by Fusarium verticillioides under different environmental conditions.

Myclobutanil is a widely used triazole fungicide, comprising two enantiomers with different fungicidal activities, non-target toxicities, and environmental fates. The enantioselective effects of myclobutanil on fumonisin B (FB) production by Fusarium verticillioides, an important pathogen, have not yet been investigated. In the present study, the fungicidal activities of rac-myclobutanil and its enantiomers on F. verticillioides cultured on maize-based media were studied under different water activity and temperature conditions. The FB levels were measured to assess the enantioselective effects on FB production when F. verticillioides were cultured treated with EC50 and EC90 concentrations (concentrations inhibiting mycelial growth by 50.0% and 90.0%, respectively) of myclobutanil and enantiomers under different conditions. The fungicidal activities of rac-myclobutanil and its enantiomers decreased with increasing temperature and decreasing water activity. Little difference in fungicidal activity was observed between the enantiomers. FB production was significantly influenced by temperature, aw, and fungicides dose. At EC50 concentrations, rac-myclobutantil and its enantiomers were shown to enhance mycotoxin production and enantioselective effects of enantiomers on FB production were observed under certain conditions. This is the first report on the differential effects of myclobutanil enantiomers on the control of F. verticillioides growth and FB production in maize-based media under different conditions.

The Fungicidal Activity of Tebuconazole Enantiomers against Fusarium graminearum and Its Selective Effect on DON Production under Different Conditions.

Tebuconazole, which consists of a pair of enantiomers with different fungicidal activities, is one of the most common fungicides used in the control of Fusarium graminearum. In this study, the fungicidal activity of rac-tebuconazole and its enantiomers against F. graminearum was determined at 0.997, 0.975, and 0.950 aw and at 20, 25, and 30 °C on wheat-based media. Then, F. graminearum was treated with rac-tebuconazole and its enantiomers at the EC10, EC50, and EC90 levels under different culture conditions, and DON production was measured. Finally, expression of the DON biosynthetic genes ( TRI5 and TRI6) was quantified by real-time RT-PCR after incubation with EC50 doses of rac-tebuconazole and its enantiomers for 4, 8, and 14 days at 30 °C and aw 0.997. The results showed that the fungicidal activity of tebuconazole was strongly influenced by temperature, aw, and the combined factors. (-)-Tebuconazole is higher in fungicidal activity than (+)-tebuconazole and rac-tebuconazole with 24-99-fold and 1.8-6.7-fold, respectively. However, (-)-tebuconazole was generally more favorable for DON production than (+)-tebuconazole under the same conditions. Additionally, (-)-tebuconazole and rac-tebuconazole induced significantly increased expression of the DON biosynthetic genes ( TRI5 and TRI6) compared to the control by the 14th day of treatment. In this research, the combination condition of 30 °C and 0.997 aw is the most suitable for DON production by F. graminearum. The test strains of F. graminearum treated with the EC10 dose of (-)-tebuconazole produced the greatest amounts of DON.

FB1-induced programmed cell death in hemocytes of Ostrinia furnacalis.

Fumonisins are a type of mycotoxin produced by Fusarium spp., mainly F. proliferatum and F. vertieilliodes. Fumonisins represent a potential hazard to the health of animals and humans. Autophagic cell death is a method of programed cell death called type II PCD, which has complicated connections with apoptosis. Our results indicated that FB1 substantially inhibited cell viability and was cytotoxic to hemocytes of Ostrinia furnacalis in a time and concentration dependent manner. We verified the activation of FB1-induced autophagy according to MDC staining, Lyso-Tracker Red probe staining, TEM observation and atg8-PE expression levels. We discovered that FB1 induced apoptosis in only a few cells based on Annexin V-FITC and PI staining. These results suggested that FB1 induced survivin inhibition and triggered autophagic cell death in the cell line. These findings might provide a plausible explanation for the underlying mechanisms of FB1 toxicity in insect cellular immune system. In addition, we provided the atg8 gene sequence of Ostrinia furnacalis, and it would be very useful to researchers if they were to study corn borer.

Over-expression of KdSOC1 gene affected plantlet morphogenesis in Kalanchoe daigremontiana.

Kalanchoe daigremontiana reproduces asexually by producing plantlets along the leaf margin. The aim of this study was to identify the function of the SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 gene in Kalanchoe daigremontiana (KdSOC1) during plantlet morphogenesis. In this study, KdSOC1 gene expression was detected at stem cell niche during in vitro somatic embryogenesis and plantlet morphogenesis. Disrupting endogenous auxin transportation suppressed the KdSOC1 gene response. Knockdown of the KdSOC1 gene caused a defect in cotyledon formation during the early heart stage of somatic embryogenesis. Over-expression (OE) of the KdSOC1 gene resulted in asymmetric plantlet distribution, a reduced number of plantlets, thicker leaves, and thicker vascular fibers. Higher KdPIN1 gene expression and auxin content were found in OE plant compared to those of wild-type plant leaves, which indicated possible KdSOC1 gene role in affecting auxin distribution and accumulation. KdSOC1 gene OE in DR5-GUS Arabidopsis reporting lines resulted in an abnormal auxin response pattern during different stages of somatic embryogenesis. In summary, the KdSOC1 gene OE might alter auxin distribution and accumulation along leaf margin to initiate plantlet formation and distribution, which is crucial for plasticity during plantlet formation under various environmental conditions.

Toxicity of the mycotoxin fumonisin B1 on the insect Sf9 cell line.

Fumonisins are a type of mycotoxin produced by Fusarium spp., mainly F. proliferatum and F. vertieilliodes, and represent a potential hazard to the health of animals and human beings. The toxicity and mechanism of action of fumonisins is ambiguous, and it is unclear whether fumonisins are toxic to insect cells. This study examines the toxicity of fumonisin B1 (FB1) and its mechanism of action in the Spodoptera frugiperda Sf9 cell line. We found that FB1 inhibited Sf9 cellular proliferation and arrested cell growth at the G2/M phase. Morphological observation showed that FB1 induced swelling, vacuole formation, and loss of adhesion in Sf9 cells. Flow cytometry analysis showed that FB1 caused depolarization of the cell membrane potential and hyperpolarization of the mitochondrial membrane potential. To uncover potential genes associated with the molecular mechanisms of FB1, 41 differentially expressed genes were identified by transcriptome analyses after FB1 treatment. These genes are putatively involved in detoxification metabolism, insect hormone regulation, cell apoptosis, and other related processes. Finally, six differentially expressed genes were chosen and validated by quantitative real-time PCR (QRT-PCR). Our test could provide a reference for other kinds of insect cells studies on FB1 stress. At the same time, our studies try to provide a possible for FB1 as a precursor compounds of biological insecticide.