Pepper vein yellow virus P0 protein triggers NbHERC3, NbBax, and NbCRR mediated hypersensitive response.

P0 proteins encoded by the pepper vein yellow virus (PeVYV) are pathogenic factors that cause hypersensitive response (HR). However, the host gene expression related to PeVYV P0-induced HR has not been thoroughly studied. Transcriptomic technology was used to investigate the host pathways mediated by the PeVYV P0 protein to explore the molecular mechanisms underlying its function. We found 12,638 differentially expressed genes (DEGs); 6784 and 5854 genes were significantly upregulated and downregulated, respectively. Transcriptomic and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) analyses revealed that salicylic acid (SA) and jasmonic acid (JA) synthesis-related gene expression was upregulated, and ethylene synthesis-related gene expression was downregulated. Ultrahigh performance liquid chromatography-tandem mass spectrometry was used to quantify SA and JA concentrations in Nicotiana benthamiana, and the P0 protein induced SA and JA biosynthesis. We then hypothesized that the pathogenic activity of the P0 protein might be owing to proteins related to host hormones in the SA and JA pathways, modulating host resistance at different times. Viral gene silencing suppression technology was used in N. benthamiana to characterize candidate proteins, and downregulating NbHERC3 (Homologous to E6-AP carboxy-terminus domain and regulator of choromosome condensation-1 dmain protein 3) accelerated cell necrosis in the host. The downregulation of NbCRR reduced cell death, while that of NbBax induced necrosis and curled heart leaves. Our findings indicate that NbHERC3, NbBax, and NbCRR are involved in P0 protein-driven cell necrosis.

Broad-spectrum nano-bactericide utilizing antimicrobial peptides and bimetallic Cu-Ag nanoparticles anchored onto multiwalled carbon nanotubes for sustained protection against persistent bacterial pathogens in crops.

Worldwide crop yields are threatened by persistent pathogenic bacteria that cause significant damage and jeopardize global food security. Chemical pesticides have shown limited effectiveness in protecting crops from severe yield loss. To address this obstacle, there is a growing need to develop environmentally friendly bactericides with broad-spectrum and sustained protection against persistent crop pathogens. Here, we present a method for preparing a nanocomposite that combines antimicrobial peptides (AMPs) and bimetallic Cu-Ag nanoparticles anchored onto multiwalled carbon nanotubes (MWCNTs). The nanocomposite exhibited dual antibacterial activity by disrupting bacterial cell membranes and splicing nucleic acids. By functionalizing MWCNTs with small AMPs (sAMPs), we achieved enhanced stability and penetration of the nanocomposite, and improved loading capacity of the Cu-Ag nanoparticles. The synthesized MWCNTs&CuNCs@AgNPs@P nanocomposites demonstrated broad-spectrum lethality against both Gram-positive and Gram-negative bacterial pathogens. Glasshouse pot trials confirmed the efficacy of the nanocomposites in protecting rice crops against bacterial leaf blight and tomato crops against bacterial wilt. These findings highlight the excellent antibacterial properties of the MWCNTs&CuNCs@AgNPs@P nanocomposite and its potential to replace chemical pesticides, offering significant advantages for agricultural applications.

Clinical characterization and outcomes of impulse oscillometry-defined bronchodilator response: an ECOPD cohort-based study.

BACKGROUND: The clinical significance of the impulse oscillometry-defined small airway bronchodilator response (IOS-BDR) is not well-known. Accordingly, this study investigated the clinical characteristics of IOS-BDR and explored the association between lung function decline, acute respiratory exacerbations, and IOS-BDR. METHODS: Participants were recruited from an Early Chronic Obstructive Pulmonary Disease (ECOPD) cohort subset and were followed up for two years with visits at baseline, 12 months, and 24 months. Chronic obstructive pulmonary disease (COPD) was defined as a post-bronchodilator forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC) ratio < 0.70. IOS-BDR was defined as meeting any one of the following criteria: an absolute change in respiratory system resistance at 5 Hz ≤ - 0.137 kPa/L/s, an absolute change in respiratory system reactance at 5 Hz ≥ 0.055 kPa/L/s, or an absolute change in reactance area ≤ - 0.390 kPa/L. The association between IOS-BDR and a decline in lung function was explored with linear mixed-effects model. The association between IOS-BDR and the risk of acute respiratory exacerbations at the two-year follow-up was analyzed with the logistic regression model. RESULTS: This study involved 466 participants (92 participants with IOS-BDR and 374 participants without IOS-BDR). Participants with IOS-BDR had higher COPD assessment test and modified Medical Research Council dyspnea scale scores, more severe emphysema, air trapping, and rapid decline in FVC than those without IOS-BDR over 2-year follow-up. IOS-BDR was not associated with the risk of acute respiratory exacerbations at the 2-year follow-up. CONCLUSIONS: The participants with IOS-BDR had more respiratory symptoms, radiographic structural changes, and had an increase in decline in lung function than those without IOS-BDR. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR1900024643. Registered on 19 July, 2019.

Residue behavior and processing factors of thirteen field-applied pesticides during the production of Chinese traditional fermented chopped pepper and chili powder.

In this study, the fate, processing factors and relationship with physicochemical properties of thirteen pesticides in field-collected pepper samples during Chinese chopped pepper and chili powder production was systematically studied. The washing, air-drying, chopping and salting and fermentation processes reduced 24.8%-62.8%, 0.9%-26.4%, 25.1%-50.3% and 16.3%-90.0% of thirteen pesticide residues, respectively, while the sun-drying processing increased the residues of eleven pesticides by 1.27-5.19 fold. The PFs of thirteen pesticides were < 1 in chopped pepper production and the PFs of eleven pesticides were more than 1 for chili powder production. The chopped pepper processing efficiency have most negative correlation with octanol-water partition coefficient. In contrast, the chili powder processing efficiency have most positive correlation with vapour pressure. Thus, this study can offer important references for assessment the pesticide residue levels in Chinese traditional fermented chopped pepper and chili powder production from fresh peppers.

Role of nuclear receptors NlHR3 and NlFTZ-F1 in regulating molting and reproduction in Nilaparvata lugens (stål).

The nuclear receptors HR3 and FTZ-F1 are highly conserved and function to regulate molting and reproduction in both hemimetabolous and holometabolous insects. However, their roles in Nilaparvata lugens are largely unknown. In the present study, we discover that NlHR3 and NlFTZ-F1 are activated in the nymph stages by ecdysone signaling. Transcription disruption of NlHR3 and NlFTZ-F1 expression prevents nymph ecdysis and metamorphosis, which leads to abnormal appearance, malformed ovaries, and lethal phenotypes. In addition, we demonstrate that NlHR3 and NlFTZ-F1 regulate molting and reproduction by interacting with the intrinsic 20E and JH signaling pathways. Our work offers a deep insight into the action mechanisms of HR3 and FTZ-F1 in insects. Moreover, NlHR3 and NlFTZ-F1 could properly be exploited as potential target genes for developing RNAi-based pesticides to control N. lugens.

Tomato chlorosis virus CPm protein is a pathogenicity determinant and suppresses host local RNA silencing induced by single-stranded RNA.

Introduction: Tomato chlorosis virus (ToCV) is a typical member of the genus Crinivirus, which severely threatens Solanaceae crops worldwide. The CPm protein encoded by ToCV has been reported to be associated with virus transmission by vectors and is involved in RNA silencing suppression, while the mechanisms remain ambiguous. Methods: Here, ToCV CPm was ectopically expressed by a Potato virus X (PVX) vector and infiltrated into Nicotiana benthamiana wild-type and GFP-transgenic16c plants. Results: The phylogenetic analysis showed that the CPm proteins encoded by criniviruses were distinctly divergent in amino acid sequences and predicted conserved domains, and the ToCV CPm protein possesses a conserved domain homologous to the TIGR02569 family protein, which does not occur in other criniviruses. Ectopic expression of ToCV CPm using a PVX vector resulted in severe mosaic symptoms followed by a hypersensitive-like response in N. benthamiana. Furthermore, agroinfiltration assays in N. benthamiana wilt type or GFP-transgenic 16c indicated that ToCV CPm protein effectively suppressed local RNA silencing induced by single-stranded but not double-stranded RNA, which probably resulted from the activity of binding double-stranded but not single-stranded RNA by ToCV CPm protein. Conclusion: Taken together, the results of this study suggest that the ToCV CPm protein possesses the dual activities of pathogenicity and RNA silencing, which might inhibit host post-transcriptional gene silencing (PTGS)-mediated resistance and is pivotal in the primary process of ToCV infecting hosts.

Impaired exercise capacity in individuals with non-obstructive small airway dysfunction.

Background: Whether individuals with non-obstructive spirometry-defined small airway dysfunction (SAD) have impaired exercise capacity is unclear, particularly in never-smokers. This study clarifies the degree of impaired exercise capacity and its potential cause in individuals with non-obstructive SAD. Methods: This community-based, multiyear cross-sectional study analyzed data collected in Guangdong, China from 2012-2019 by the National Science and Technology Support Plan Program. Measurements of exercise capacity [peak work rate and peak oxygen uptake ( V ˙ O 2peak )] in participants with non-obstructive spirometry-defined SAD (n=157) were compared with those in controls (n=85) and Global Initiative for Chronic Obstructive Lung Disease (GOLD) I patients (n=239). Subgroup analyses were performed by smoking status. Results: The risk of impaired exercise capacity was significantly higher in participants with non-obstructive SAD [ V ˙ O 2peak <84%predicted, adjusted odds ratio (aOR) =2.53; 95% confidence interval (CI): 1.42-4.52] than in controls but was not significantly different from that in GOLD I patients. Results were consistent within subgroups of smoking status (ever-smokers: non-obstructive SAD vs. controls, aOR =2.44; 95% CI: 1.08-5.51; never-smokers: non-obstructive SAD vs. controls, aOR =2.38, 95% CI: 1.02-5.58). Participants with non-obstructive SAD had a significantly lower peak work rate (ß=-10.5; 95% CI: -16.3 to -4.7) and V ˙ O 2peak (%predicted, ß=-4.0; 95% CI: -7.7 to -0.2) and tended to have higher ventilatory equivalents for carbon dioxide at the ventilatory threshold ( V ˙ E / V ˙ CO 2AT , ß=1.1; 95% CI: -0.1 to 2.3) when compared with controls. Both peak work rate and V ˙ O 2peak were negatively correlated with V ˙ E / V ˙ CO 2AT . Conclusions: Although not meeting the current criteria for chronic obstructive pulmonary disease, individuals with non-obstructive SAD have impaired exercise capacity that may be associated with ventilatory inefficiency regardless of smoking status.

CRISPR/Cas9-mediated gene editing of vacuolar ATPase subunit d mediates phytohormone biosynthesis and virus resistance in rice.

Vacuolar ATPases (V-ATPases) are proton pumps for proton translocation across membranes that utilize energy derived from ATP hydrolysis; OsV-ATPase subunit d (OsV-ATPase d) is part of an integral, membrane-embedded V0 complex in the V-ATPase complex. Whether OsV-ATPase d is involved in phytohormone biosynthesis and resistance in rice remains unknown. The knockout mutants of OsV-ATPase d in rice were generated using the CRISPR/Cas9 system, and mutation of OsV-ATPase d did not show any detrimental effect on plant growth or yield productivity. Transcriptomic results showed that OsV-ATPase d is probably involved in mediating the biosynthesis of plant hormones and resistance in rice. Compared to wild type, mutation of OsV-ATPase d significantly increased JA and ABA biosynthesis and resistance against Southern rice black-streaked dwarf virus (SRBSDV), but it decreased resistance against Rice stripe virus (RSV) in rice. The data presented in this study reveal that OsV-ATPase d mediates phytohormone biosynthesis and virus resistance in rice and can be selected as a potential target for resistance breeding in rice.

Chlorpyrifos residue level and ADHD among children aged 1-6 years in rural China: A cross-sectional study.

Background: Attention-deficit/hyperactivity disorder (ADHD) is one of the most common neurodevelopmental disorders in childhood and is caused by both genetic and environmental factors. As genetic factors are nonmodifiable, environmental factors have attracted increasing attention. Objective: To investigate the relationships between urinary chlorpyrifos (CPF) levels, blood micronutrient levels, and ADHD prevalence in children living in rural areas of China. Methods: This cross-sectional study collected data on CPF exposure (according to urinary levels), blood micronutrient levels, and ADHD prevalence in children aged 1-6 years in rural China. The CPF levels were determined by mass spectrometry. Blood levels of micronutrients, including zinc, iron, calcium, copper, magnesium, and vitamin D, were measured by professional detection kits. ADHD was diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition. Descriptive statistics and univariate analysis were conducted using SPSS 21.0, and path analysis was conducted using Mplus 8.0. Results: Of the 738 children who met the eligibility criteria, 673 children (673/738, 91.2%) were included in the final analysis. Baseline questionnaires and urine samples were collected from all 673 subjects. A total of 672 children provided blood samples for micronutrient testing, and 651 completed the ADHD assessment. Approximately one-fifth of children (144/673, 21.4%) had detectable levels of CPF in their urine, and 6.9% (45/651) were diagnosed with ADHD. Path analysis showed that the total effect of CPF exposure on ADHD risk was 0.166 (P < 0.05), with a direct effect of 0.197 (P < 0.05) and an indirect effect of -0.031 (P < 0.05) via vitamin D. The mediating effect of urinary CPF levels on ADHD risk via vitamin D was 18.67%. Conclusion: Higher levels of CPF exposure are associated with higher risk of ADHD. Additionally, increasing vitamin D levels may have a beneficial effect on the relationship between CPF exposure and ADHD risk. Our findings highlight the importance of modifying environmental factors to reduce ADHD risk and provide insight into future ADHD interventions.

Residue analysis and dietary risk assessment of tebuconazole in loquat and sugarcane after open-field application in China.

Tebuconazole is an effective but potentially carcinogenic fungicide. Tebuconazole residues in loquat and sugarcane were investigated, together with an analysis of their dietary risk. Solid-phase extraction and high-performance liquid chromatography-tandem mass spectrometry showed fortified recoveries of 87% to 98%, with standard deviations ≤ 6.4%. The tebuconazole residues were below 6.34 mg kg-1 for whole loquat and 0.80 mg kg-1 for sugarcane. Based on the final residue test, the chronic hazard index was 90.74% in whole loquat and sugarcane, while the acute hazard index ranged from 66.32% to 109.49% for loquat and 33.37% to 76.93% for sugarcane. The assessment of potential dietary risk showed that while loquats may represent an acute risk to human health, whereas sugarcane consumption was safe. A 21-day pre-harvest interval is proposed to comply with maximum residue limit regulations and the observed risks when using 430 g/L tebuconazole for treating sugarcane. These findings offer guidance for tebuconazole formulation and use.

Monitoring residue levels and dietary risk assessment of thiamethoxam and its metabolite clothianidin for Chinese consumption of Chinese kale.

BACKGROUND: Thiamethoxam is widely used to control pests in Chinese kale, popularly consumed leafy vegetables. The potential risk to the environment and human health has aroused much public concern. Therefore, it is important to investigate the degradation behavior, residue distribution and dietary risk assessment of thiamethoxam in Chinese kale. RESULTS: A sensitive analytical method for determination of thiamethoxam and its metabolite clothianidin residue in Chinese kale was established and validated through a quick, easy, cheap, effective, rugged, and safe (QuEChERS) technique with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The recoveries were 85.4-101.2% for thiamethoxam and 79.5-108.1% for clothianidin, with the relative standard deviations (RSDs) of 0.9-10.2% and 1.8-6.0%, respectively. For the dissipation kinetics, the data showed that thiamethoxam in Chinese kale was degraded with the half-lives of 4.1 to 4.5 days. In the terminal residue experiments, the residues of thiamethoxam were 0.017-0.357 mg kg-1 after application 2-3 times with a preharvest interval (PHI) of 7 days under the designed dosages. The chronic and acute dietary exposure assessment risk quotient (RQ) values of thiamethoxam in Chinese kale for different Chinese consumers were 0.08-0.19% and 0.05-0.12%, respectively, and those of clothianidin were 0.01-0.04% and 0.02-0.04%, respectively, all of the RQ values were lower than 100%. CONCLUSION: Thiamethoxam in Chinese kale was rapidly degraded following first-order kinetics models. The dietary risk of thiamethoxam and clothianidin through Chinese kale was negligible to consumers. The results from this study are important reference for Chinese governments to developing criteria for the safe and rational use of thiamethoxam, setting maximum residue levels (MRLs), monitoring the quality safety of agricultural products and protecting consumer health. © 2021 Society of Chemical Industry.

Residue changes and processing factors of eighteen field-applied pesticides during the production of Chinese Baijiu from rice.

The fate of eighteen pesticides in field-collected rice samples during Chinese Baijiu production was systematically studied. The results indicated that steeping decreased flonicamid residue by 73.2% due to its high water-solubility and low octanol/water partition coefficient. The steaming step reduced pesticide residues by 32.0%-75.3% through evaporation or thermal degradation. After steaming, the pesticide residues were further reduced by 39.8-74.2% in fermentation which might be caused by biological degradation. In addition, distillation was shown to be most effective, responsible for greater than 90% losses of the remaining pesticide residues. The processing factors (PFs) were generally lower than 1 for different processes and the whole procedure. These results revealed that the procedure of Chinese Baijiu production could dramatically decrease residues of all the eighteen pesticides. Overall, this study provide important references for monitoring pesticide residue levels during the production of Chinese Baijiu from rice, and ensuring proper risk assessment from pesticide contamination.

Residue behavior, transfer and risk assessment of tolfenpyrad, dinotefuran and its metabolites during tea growing and tea brewing.

BACKGROUND: Tolfenpyrad and dinotefuran are two representative pesticides used for pest control in tea gardens. Their application may bring about a potential risk to the health of consumers. Therefore, it is essential to investigate the residue behavior, transfer and risk assessment of tolfenpyrad, dinotefuran and metabolites from tea garden to teacup. RESULTS: An effective analytical method was established and validated to simultaneously determine tolfenpyrad, dinotefuran and its metabolites (DN and UF) in tea. The average recoveries of tolfenpyrad, dinotefuran, DN and UF were in the range 72.1-106.3%, with relative standard deviations lower than 11.8%. On the basis of the proposed method, the dissipation of tolfenpyrad and dinotefuran in fresh tea leaves followed first-order kinetics models with half-lives of 4.30-7.33 days and 4.65-5.50 days, respectively. With application amounts of 112.5-168.75 g a.i. ha-1 once or twice, the terminal residues of tolfenpyrad and total dinotefuran in green tea were lower than 19.6 and 7.13 mg kg-1 , respectively, and below their corresponding maximum residue limits . The leaching rates of tolfenpyrad and total dinotefuran during the tea brewing were in the ranges 1.4-2.3% and 93.7-98.1%, respectively. CONCLUSION: Tolfenpyrad and dinotefuran in tea were easily degraded. The RQc and RQa values for tolfenpyrad were 37.6% and 5.4%, which were much higher than for dinotefuran at 24.7% and 0.84%, respectively. The data indicated that there was no significant health risk in tea for consumers at the recommended dosages. The results provide scientific data regarding the reasonable use of tolfenpyrad and dinotefuran aiming to ensure safe tea consuption. © 2021 Society of Chemical Industry.

Study on degradation behaviour, residue distribution, and dietary risk assessment of propiconazole in celery and onion under field application.

BACKGROUND: Propiconazole is widely used to control fungal diseases in field crops, including celery and onion. The potential risk to the environment and human health has aroused much public concern. Therefore, it is significant to investigate the degradation behaviour, residue distribution, and dietary risk assessment of propiconazole in celery and onion. RESULTS: A sensitive analytical method for determination of propiconazole residue in celery and onion was established and validated through high-performance liquid chromatography tandem mass spectrometry. The average recovery rate of propiconazole ranged from 85.7% to 101.8%, with a relative standard deviation of 2.1-6.3%. For the dissipation kinetics, the data showed that propiconazole in celery and onion was degraded, with half-lives of 6.1-6.2 days and 8.7-8.8 days respectively. In the terminal residue experiments, the residues of propiconazole were below 4.66 mg kg-1 in celery after application two or three times and were below 0.029 mg kg-1 in onion after application of three or four times with an interval of 14 days under the designed dosages. The chronic and acute dietary exposure assessments for propiconazole were valued by risk quotient, with all values being lower than 100%. CONCLUSION: Propiconazole in celery and onion was rapidly degraded following first-order kinetics models. The dietary risk of propiconazole through celery or onion was negligible to consumers. The study not only offers a valuable reference for reasonable usage of propiconazole on celery and onion, but also facilitates the establishment of maximum residue limits in China. © 2020 Society of Chemical Industry.

First evidence showing that Pepper vein yellows virus P4 protein is a movement protein.

BACKGROUND: Plant viruses move through plasmodesmata (PD) to infect new cells. To overcome the PD barrier, plant viruses have developed specific protein(s) to guide their genomic RNAs or DNAs to path through the PD. RESULTS: In the present study, we analyzed the function of Pepper vein yellows virus P4 protein. Our bioinformatic analysis using five commonly used algorithms showed that the P4 protein contains an transmembrane domain, encompassing the amino acid residue 117-138. The subcellular localization of P4 protein was found to target PD and form small punctates near walls. The P4 deletion mutant or the substitution mutant constructed by overlap PCR lost their function to produce punctates near the walls inside the fluorescent loci. The P4-YFP fusion was found to move from cell to cell in infiltrated leaves, and P4 could complement Cucumber mosaic virus movement protein deficiency mutant to move between cells. CONCLUSION: Taking together, we consider that the P4 protein is a movement protein of Pepper vein yellows virus.

The NIa-Protease Protein Encoded by the Pepper Mottle Virus Is a Pathogenicity Determinant and Releases DNA Methylation of Nicotiana benthamiana.

It is well documented that the canonical function of NIa-protease (NIa-Pro) of the potyviruses is responsible for cleaving the viral polyprotein into functional proteins. Although NIa-Pro is vital for the infection cycle of potyviruses, the function of NIa-Pro in the interaction of the potyvirus host is not clear. In this study, NIa-Pro is ectopically expressed from a potato virus X (PVX) vector and infiltrates Nicotiana benthamiana wild type and 16-TGS. The pathogenicity and inhibition of host transcriptional gene silencing (TGS) are characterized. Ectopic expression of NIa-Pro from a PVX vector resulted in severe mosaic symptoms followed by a hypersensitive-like response in N. benthamiana. Furthermore, PepMoV NIa-Pro was able to reverse established TGS of a green fluorescent protein transgene by reducing methylation of promoter sequences in N. benthamiana and possessed the capacity to interfere with the global methylation of N. benthamiana. Taken together, the results of this study likely suggest that PepMoV NIa-Pro is a pathogenicity determinant and a potent suppressor of host TGS and suggest that NIa-Pro may employ novel mechanisms to suppress host antiviral defenses. To the best of our knowledge, this is the first report of a plant RNA virus modulating host TGS in a novel manner by interfering with the establishment of the methylation step of the plant DNA methylation pathway.

Knowledge and behavior regarding pesticide use: a survey among caregivers of children aged 1-6 years from rural China.

Little is known about pesticide exposure risks for children in rural areas in China, many of whom have been left behind by migrant workers. To survey caregivers of children in rural China and assess their pesticide use and disposal methods, the measures were used to protect the children and their perceptions of the adverse effects of pesticides on children's health. Three or four villages in each of Xinhua, Gongan and Sansui Counties in China were selected by random cluster sampling. The main caregivers of children aged 1-6 years were surveyed in face-to-face interviews. The questionnaire used was adapted from the World Health Organization "Exposure to Pesticides: Standard Protocol" survey and similar studies. The study included 464 caregivers (mean age, 46.4 years), who were most commonly the children's grandparents (65.3%). Among the caregivers, 41.9% were educated to middle school level or higher, 45.4% had a household income < 297 USD, and 29.7% had received education/training about pesticide use/adverse health effects in children. The score for caregivers' knowledge of the adverse effects of pesticides on children's health was higher in those who had received education/training (5.1 ± 2.6 vs. 3.4 ± 2.5, P < 0.001). Factors associated with unsafe behaviors during pesticide use included grandparent as the caregiver (odds ratio [OR] 0.551; 95% confidence interval [95% CI] 0.368-0.824; P = 0.004), annual income < 297 USD (OR 0.580; 95% CI 0.395-0.853; P = 0.006), and insufficient health-related education/training (OR 0.436; 95% CI 0.286-0.665; P < 0.001). Improved education and training are needed to promote the safe use of pesticides by caregivers of children in rural China.

Biodegradation of fenpropathrin by Rhodopseudomonas sp. strain PSB07-21 cultured under three different growth modes.

This study aimed at the biodegradation of fenpropathrin by Rhodopseudomonas sp. strain PSB07-21 cultured under different growth modes. The biomass production, cell surface hydrophobicity and fenpropathrin biodegradation efficiency of the strain PSB07-21 cultured under the photoheterotrophic growth mode were better than that shown by the strain PSB07-21 cultured under the photoautotrophic or the chemotrophic growth mode. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis using cell-free protein extracts showed several distinct protein bands in the gels representing the strain PSB07-21 cultured under the photoheterotrophic growth mode. The fenpropathrin enzymatic degradation was clearly affected the bacterial growth mode. Results obtained from this study should improve our knowledge regarding fenpropathrin biodegradation under field conditions. Our findings can also be used to optimize the usage of Rhodopseudomonas sp. PSB07-21 in field applications.

Adaptation mechanism and tolerance of Rhodopseudomonas palustris PSB-S under pyrazosulfuron-ethyl stress.

BACKGROUND: Pyrazosulfuron-ethyl is a long lasting herbicide in the agro-ecosystem and its residue is toxic to crops and other non-target organisms. A better understanding of molecular basis in pyrazosulfuron-ethyl tolerant organisms will shed light on the adaptive mechanisms to this herbicide. RESULTS: Pyrazosulfuron-ethyl inhibited biomass production in Rhodopseudomonas palustris PSB-S, altered cell morphology, suppressed flagella formation, and reduced pigment biosynthesis through significant suppression of carotenoids biosynthesis. A total of 1127 protein spots were detected in the two-dimensional gel electrophoresis. Among them, 72 spots representing 56 different proteins were found to be differently expressed using MALDI-TOF/TOF-MS, including 26 up- and 30 down-regulated proteins in the pyrazosulfuron-ethyl-treated PSB-S cells. The up-regulated proteins were involved predominantly in oxidative stress or energy generation pathways, while most of the down-regulated proteins were involved in the biomass biosynthesis pathway. The protein expression profiles suggested that the elongation factor G, cell division protein FtsZ, and proteins associated with the ABC transporters were crucial for R. palustris PSB-S tolerance against pyrazosulfuron-ethyl. CONCLUSION: Up-regulated proteins, including elongation factor G, cell division FtsZ, ATP synthase, and superoxide dismutase, and down-regulated proteins, including ALS III and ABC transporters, as well as some unknown proteins might play roles in R. palustris PSB-S adaptation to pyrazosulfuron-ethyl induced stresses. Functional validations of these candidate proteins should help to develope transgenic crops resistant to pyrazosulfuron-ethyl.

Cloning and characterization of a pyrethroid pesticide decomposing esterase gene, Est3385, from Rhodopseudomonas palustris PSB-S.

Full length open reading frame of pyrethroid detoxification gene, Est3385, contains 963 nucleotides. This gene was identified and cloned based on the genome sequence of Rhodopseudomonas palustris PSB-S available at the GneBank. The predicted amino acid sequence of Est3385 shared moderate identities (30-46%) with the known homologous esterases. Phylogenetic analysis revealed that Est3385 was a member in the esterase family I. Recombinant Est3385 was heterologous expressed in E. coli, purified and characterized for its substrate specificity, kinetics and stability under various conditions. The optimal temperature and pH for Est3385 were 35 °C and 6.0, respectively. This enzyme could detoxify various pyrethroid pesticides and degrade the optimal substrate fenpropathrin with a Km and Vmax value of 0.734 ± 0.013 mmol·l-1 and 0.918 ± 0.025 U·µg-1, respectively. No cofactor was found to affect Est3385 activity but substantial reduction of enzymatic activity was observed when metal ions were applied. Taken together, a new pyrethroid degradation esterase was identified and characterized. Modification of Est3385 with protein engineering toolsets should enhance its potential for field application to reduce the pesticide residue from agroecosystems.