Domestication and selection footprints in Persian walnuts (Juglans regia).

Walnut (Juglans) species are economically important hardwood trees cultivated worldwide for both edible nuts and high-quality wood. Broad-scale assessments of species diversity, evolutionary history, and domestication are needed to improve walnut breeding. In this study, we sequenced 309 walnut accessions from around the world, including 55 Juglans relatives, 98 wild Persian walnuts (J. regia), 70 J. regia landraces, and 86 J. regia cultivars. The phylogenetic tree indicated that J. regia samples (section Dioscaryon) were monophyletic within Juglans. The core areas of genetic diversity of J. regia germplasm were southwestern China and southern Asia near the Qinghai-Tibet Plateau and the Himalayas, and the uplift of the Himalayas was speculated to be the main factor leading to the current population dynamics of Persian walnut. The pattern of genomic variation in terms of nucleotide diversity, linkage disequilibrium, single nucleotide polymorphisms, and insertions/deletions revealed the domestication and selection footprints in Persian walnut. Selective sweep analysis, GWAS, and expression analysis further identified two transcription factors, JrbHLH and JrMYB6, that influence the thickness of the nut diaphragm as loci under selection during domestication. Our results elucidate the domestication and selection footprints in Persian walnuts and provide a valuable resource for the genomics-assisted breeding of this important crop.

N-Doped Carbon Dots for Selective Detection of Fe3+ and Degradation of Fe3+/Basic Red 9 Complexes in Water Samples.

In this work, the eco-friendly N-doped carbon dots KF-CDs and A-CDs were derived from kiwifruit by a simple one-step hydrothermal strategy at 180 °C for 6 h. KF-CDs have a high fluorescence quantum yield (27.85%), it is obviously rapid quenched by Fe3+, and have a good linear relationship from 1 to 8.26 µM (the detection limit was 0.077 µM). Basic red 9 is extensively used in biological, environmental and industry. Although it makes a great contribution to the economy, its toxicity should be taken seriously, especially with harmful metal ions. Within 2 h, A-CDs could degrade basic red 9 with degradation efficiency 89.6%, even though there was a stable compound formed with Fe3+ that the degradation efficiency was up to 88.3%. The results complement the research blank of carbon dots in catalytic degradation of basic red 9.

Multi-omics analyses reveal the responses of wheat (Triticum aestivum L.) and rhizosphere bacterial community to nano(micro)plastics stress.

The pervasive existence of nanoplastics (NPs) and microplastics (MPs) in soil has become a worldwide environmental concern. N/MPs exist in the environment in a variety of forms, sizes, and concentrations, while multi-omics studies on the comprehensive impact of N/MPs with different properties (e.g. type and size) on plants remain limited. Therefore, this study utilized multi-omics analysis methods to investigate the effects of three common polymers [polyethylene-NPs (PE-NPs, 50 nm), PE-MPs (PE-MPs, 10 µm), and polystyrene-MPs (PS-MPs, 10 µm)] on the growth and stress response of wheat, as well as the rhizosphere microbial community at two concentrations (0.05 and 0.5 g/kg). PS and PE exhibited different effects for the same particle size and concentration. PE-NPs had the most severe stress effects, resulting in reduced rhizosphere bacteria diversity, plant biomass, and antioxidant enzyme activity while increasing beneficial bacteria richness. N/MPs altered the expression of nitrogen-, phosphorus-, and sulfur-related functional genes in rhizosphere bacteria, thereby affecting photosynthesis, as well as metabolite and gene levels in wheat leaves. Partial least squares pathway models (PLSPMs) indicated that concentration, size, and type play important roles in the impact of N/MPs on the plant ecological environment, which could have essential implications for assessing the environmental risk of N/MPs.

Multivariate response surface methodology assisted modified QuEChERS method for the rapid determination of 39 pesticides and metabolites in medlar.

A modified QuEChERS method coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was established for residue analysis of 39 pollutants (34 commonly used multi-class pesticides and 5 metabolites) in medlar matrices (fresh, dried, and medlar juice). Samples were extracted using water with 0.1 % formic acid: acetonitrile (5: 10, v/v). The phase-out salts and five different cleanup sorbents (including N-propyl ethylenediamine (PSA), octadecyl silane bonded silica gel (C18), graphitized carbon black (GCB), Carbon nanofiber (C-Fiber) and MWCNTs) were investigated to improve the purification efficiency. The Box-Behnken Design (BBD) study was employed for an optimal solution of the volume of extraction solvent, phase-out salt, and the purification sorbents for the analytical method. The average recoveries of the target analytes in the three medlar matrices ranged from 70 % to 119 % with relative standard deviations (RSDs) of 1.0 %-19.9 %. Screening of market samples (fresh and dried medlars) collected from the major producing regions in China showed that 15 pesticides and metabolites were detected in the samples at concentrations of 0.01-2.22 mg/kg, and none of which exceeded the maximum residue limits (MRLs) set in China. The results showed that the risk of food safety by consumption of medlar products caused by the use of pesticides was low. The validated method could be used for rapid and accurate screening of multi-class multi-pesticide residues in Medlar for food safety.

Method validation, residue analysis and dietary risk assessment of trifloxystrobin and trifloxystrobin acid in milk, eggs and pork.

Trifloxystrobin (TFS) is a widely used strobilurin fungicide and its residues accumulating in animal-derived food could result in potential harm to consumers. By optimization of extraction solvents and cleanup sorbents, a residue analysis method for TFS and its metabolite trifloxystrobin acid (TFSA) was established in milk, eggs and pork based on QuEChERS sample preparation and LC-MS/MS. The calibration curves exhibited good linearity with determination coefficients (R2 ) >0.9930 over the range of 0.5-250 ng/ml for both TFS and TFSA. The recoveries of the two analytes were 81-100% with RSD 3-10% and 76-96% with RSD 2-13%, respectively. The limit of quantification (LOQ) was 1 ng/g for both analytes. The milk, egg and pork samples, 30 each, were collected from the 30 main producing regions in China, and residues of TFS and TFSA were analyzed. The concentrations of both analytes were lower than the corresponding LOQs and maximum residue limits. Long-term dietary risk assessment showed that the hazard quotients were 0.001-0.003%, indicating an absence of unacceptable risks in milk, eggs and pork to the health of common consumers in China.

Method validation and residue analysis of methoxyfenozide and metaflumizone in Chinese broccoli under field conditions by liquid chromatography with tandem mass spectrometry.

Methoxyfenozide and metaflumizone are insecticides used on Chinese broccoli to prevent insects and increase yield. However, the residues are potentially harmful to the environment and consumers. In this study, the quick, easy, cheap, effective, rugged, safe method with high-performance liquid chromatography with tandem mass spectrometry was modified and validated for determination of methoxyfenozide and metaflumizone in Chinese broccoli. The clean-up efficiency of different sorbents including C18 , primary secondary amine, graphitized carbon black, and carbon nanofiber was compared. Recoveries of the validated method were 71.8-94.6% with relative standard deviations of 1.5-3.2% and the limits of quantification were 0.01 and 0.005 mg/kg for methoxyfenozide and metaflumizone, respectively. A storage stability test showed almost no degradation of methoxyfenozide in Chinese broccoli, however, the degradation rate of metaflumizone was 22.9% after 10-wk storage at -20°C. In field trials in four producing regions, the dissipation of both methoxyfenozide and metaflumizone in Chinese broccoli was fast, with half-lives of only 1.0-5.1 and 0.7-2.5 days, respectively. Terminal residues after application of the two pesticides were all below 1.0 mg/kg after 5 days.

Hydrolysis Kinetics of a Novel 3,4-Dichloroisothiazole Plant-Activator Candidate LY5-24-2 by UPLC-Q-TOF.

Hydrolysis characteristics of a novel 3,4-dichloroisothiazole based fungicide with activating plant defense responses as a candidate plant-activator LY5-24-2 were investigated under different conditions (pH and temperature) using ultra-performance liquid chromatography (UPLC) and quadrupole Time-of-Flight (Q-TOF). The hydrolysis case complied with the first-order kinetic model, with half-lives ranging from 4.8 h to 3.2 days at pH 4, 7, 9 and temperature at 25 and 50℃. One of the hydrolysis metabolite 3,4-dichloroisothiazole-5-carboxylic acid (metabolite 1, M1) was determined and quantified using authentic standard. The other hydrolysate 3-chloro-5-(trifluoromethyl) pyridin-2-amine (metabolite 2, M2) was determined and identified according to accurate mass information, fragmentation patterns and principle component analysis (PCA). By utilizing high-resolution mass spectrometry and multivariate statistical analysis, hydrolysis dynamic of the metabolites was characterized and figured out. This research provided a non-target screening method to analyze hydrolysis metabolites of a new plant-activator and to find its degradation products in aqueous solution.

Environmental behavior and influencing factors of glyphosate in peach orchard ecosystem.

In this paper, several experiments were carried out to study the environmental behavior and influencing factors of glyphosate (PMG) in peach orchard ecosystem. The results of field experiments showed that PMG and its metabolite aminomethylphosphonic acid (AMPA) were detected in peach tree leaves and peach tree fruits, although PMG was only sprayed on the soil. The residues of PMG and AMPA in peach tree leaves were ~0.1 mg/kg and ~0.5 mg/kg and in peach tree fruits were ~0.01 mg/kg and 0.07-0.11 mg/kg, respectively. By conducting a series of laboratory simulation experiments, the environmental factors affecting the degradation of PMG were screened and evaluated. The results showed that PMG metabolized much faster in loess soil than red soil and black soil (with the DT50 of 11.6 days, 62.4 days, and 34.1 days, respectively). By analyzing the basic properties of the soil, we investigated the effects of pH, moisture content, organic matter (exogenous biochar) and ambient temperature using orthogonal experiments, and the results were further confirmed by microbial experiment. The results showed that alkaline conditions (pH = 7.8/9), high water content (25%) and microorganisms could promote the degradation of PMG. Sterile soil environment had a negative impact on the metabolic behavior of PMG to AMPA.

Residue determination of triclopyr and aminopyralid in pastures and soil by gas chromatography-electron capture detector: Dissipation pattern under open field conditions.

In this study, a new method for the simultaneous quantitative determination of triclopyr and aminopyralid in forage grass, hay, and soil was developed and validated using gas chromatography coupled with electron capture detector (GC-ECD). In this method, a simple and maneuverable esterification reaction was applied to convert the two acidic herbicides into their ester form with methanol. The target compounds were extracted with 1% hydrochloric acid-acetonitrile, esterified, purified by florisil solid-phase extraction cartridge, and detected in a single run by the GC-ECD. The average recoveries using this method, at different fortified levels, ranged from 80% to 104% with intra-day and inter-day RSDs in the range of 1.2-10.8% and 3.3-10.3% for both the herbicides, respectively. The LODs were below 0.02 mg/kg while the LOQs were below 0.05 mg/kg, both of which were much lower than the maximum residue limits (MRLs) of 25-700 mg/kg in pastures, as established by the USA (the code of federal regulations). The open field dissipation and residual analysis in pastures and soil were conducted with the commercial formulation at two locations. With time, both triclopyr and aminopyralid dissipated via first-order kinetics. In forage grass, both compounds degraded rapidly over the first 14- or 21-d period and at a slow rate over the remainder of experimental days. In soil, they degraded at a relatively slow rate, and dissipated steadily to below or close to the LOQ by 60-d post application. The half-lives of triclopyr were 1.4-1.8 d and 6.2-9.0 d and aminopyralid were 1.7-2.1 d and 8.2-10.6 d in terms of forage grass and soil, respectively. The terminal residue results indicated that on 7 d after the treatment, the residues of aminopyralid and triclopyr in forage grass and hay were lower than the MRLs set by the USA. This work can provide guidance on the reasonable use of these herbicides and also provide an analytical method for the determination of triclopyr and aminopyralid in pasture and soil.

Dissipation, residues, and evaluation of processing factor for spirotetramat and its formed metabolites during kiwifruit growing, storing, and processing.

Spirotetramat is widely used around the world to control sucking pests and may form in agricultural products. In the current study, the dissipation, residues, and evaluation of processing factor (PF) for spirotetramat and its formed metabolites were investigated during kiwifruit growing, storing, and processing. The residue analysis method was established based on high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) combined with a QuEChERS method to detect the residues of spirotetramat and its metabolites in kiwifruit and its processed products. The method provided recoveries of 74.7-108.7%, and the relative standard deviations (RSDs) were 0.6-13.1%. The LOQs of spirotetramat and its four metabolites were 1 µg kg-1. The degradation of spirotetramat was best fitted for the first-order kinetics model with a half-life of 9.90-10.34 days in the field and 24.75-30.13 days during storage. Residues of spirotetramat and its formed metabolites in kiwifruit would not pose dietary risk to consumers. Moreover, the peeling and fermentation were the highest removal efficiency for the spirotetramat and its formed metabolite residues during processing. The PF values calculated after each individual process were < 1, indicating a significant reduction of residues in different processing processes of kiwifruit. The spirotetramat was degraded during kiwifruit wine-making process with half-lives of 3.36-4.91 days. B-enol and B-keto were the main metabolites detected in kiwifruit and its processed products. This study revealed the residues of spirotetramat and its formed metabolites in kiwifruit growing, storing, and processing, which helps provide reasonable data for studying the dietary risk factors of kiwifruits and products.

Effects of storage and processing on the residual distribution and behavior of five preservatives and their metabolites in pomegranate.

Pomegranate are often treated with preservatives during storage. This study investigated the effects of storage and food processing on the residual behavior of the five commonly used preservatives (prochloraz, thiophanate-methyl, pyrimethanil, imazalil, and difenoconazole) and their metabolites in pomegranate and its products. The LOQs for all target compounds were 0.001 mg kg-1. The residue levels of five preservatives in the calyx was highest, followed by the peel, stalk, septum, umbilicus, and seed. For the migration ability, the five preservatives from pomegranate peel to seed was negatively correlated with their octanol/water partition coefficients. The processing factors of each procedures of juice, wine, vinegar, and pectin processing were <1. Nevertheless, the PF values in drying peel during the overall process ranged from 1.26 to 4.09. Hence, it is worth noting that consumption of pomegranate essential oil and drying peel may pose a potential risk to the health of consumers.

Dissipation behavior and risk assessment of imidacloprid and its metabolites in apple from field to products.

Pesticides play vital roles in controlling pests and boosting crop yields. Imidacloprid is widely used all over the world and may form in agricultural products. The presence of pesticide residues in apples raises serious health concerns. Understanding the residual fate of imidacloprid is critical for food safety and human health. In this study, the dissipation behavior, metabolism, household processing and risk assessment of imidacloprid and its metabolites in apple were investigated from filed to products. Field experiment results suggested that the half-lives of imidacloprid at 5 times the recommended dosage was 1.5 times that of the standard dosage. And the final residues of imidacloprid were less than the established maximum residue limits (MRLs). Clarification and simmering had little effect on the reduction the residues of imidacloprid and its metabolites. The calculated processing factors were lower than 1 for imidacloprid and its metabolites, implying that the residual ratios of imidacloprid and its metabolites in each steps of the food processing were reduced. The risk quotients were <1 for all Chinese people, indicating that acceptable risks associated with dietary exposure to imidacloprid in apple. However, the higher risks were observed in young people than adults, and females faced higher risks than males. Given high residue levels in pomace, imidacloprid and its metabolites should be further studied in commercial byproducts.

Key metabolites and mechanistic insights in forchlorfenuron controlling kiwifruit development.

Forchlorfenuron (CPPU) is a plant growth regulator widely applied on kiwifruit to improve yield, however, there are rarely reports on its effects on the nutrients of kiwifruits. Based on UHPLC-Q-TOF-MS, the effects of CPPU on metabolism profile and nutrient substances of two kiwifruit varieties during development were investigated by non-targeted metabolomics. A total of 115 metabolites were identified, and 29 differential metabolites were confirmed and quantified using certified reference standards. Metabolic profile indicated that CPPU promoted kiwifruit development during the main expansion stages at the molecular level, and the effects varied slightly for different varieties. In the early and middle stages of kiwifruit development, the anthocyanin, flavone and flavonol biosynthesis were down-regulated in both varieties, and flavanols biosynthesis was down-regulated only in Hayward variety. Arginine biosynthesis was down-regulated at all stages till the harvest. Although the synthesis of these nutrient substances in kiwifruits was mostly down-regulated by CPPU, the negative effects became mild at harvest time, and positively, the significant increase of sucrose and decrease of organic acids at harvest time could help to improve the taste of kiwifruits.

Determination of mepiquat chloride in cotton crops and soil and its dissipation rates.

A simple, accurate and highly sensitive analytical method was developed in this study for determining the residues and dissipation dynamics of mepiquat chloride in soil and cotton crops (including plant and seed). The samples in this method were directly extracted with methanol-ammonium acetate solution and followed by analysis of high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The intra-day and inter-day recoveries of mepiquat chloride were in the range of 76.58-98.87 percent and 73.48-104.7 percent with relative standard deviations (RSDs) of 0.89-5.04 percent and 3.84-10.7 percent, respectively. The limit of quantification (LOQ) was 0.05mgkg(-1) for soil and 0.1mgkg(-1) for cotton plant and seed. The half-life of mepiquat chloride in cotton plants and in soil was 2.51-3.85 days and 7.56-10.50 days, respectively. The final residues of mepiquat chloride in all cotton seeds were below the MRLs of EU (5mgkg(-1)), Japan and America (2mgkg(-1)) over 14 days after the last spraying, and were below 1mgkg(-1) (the MRLs set by Argentina and Australia) over 21 days after the last spraying at recommended dosage. The final residues in soil were below the LOD (0.01mgkg(-1)) over 21 days after the last spraying.

The fate of thiamethoxam and its main metabolite clothianidin in peaches and the wine-making process.

Thiamethoxam is widely used to control a large number of insect pests of peach crops. Understanding the fate of thiamethoxam and its main metabolite clothianidin in field peach, during storage, and in the processing of peach wine is of vital importance for food safety. The thiamethoxam and clothianidin were separated and determined by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). Field and storage results showed that the half-lives of thiamethoxam were 4.9-5.5 and 10.3-15.8 days, respectively. The processing factors (PFs) of all the wine-making procedures were less than 1, and the PFs of the overall process ranged from 0.10 to 0.47. The highest elimination rate was obtained for thiamethoxam and clothianidin during the fermentation process. The results from this study could understand the dissipation kinetics and residual levels of thiamethoxam and clothianidin in peach, and also help to accurately assess their risks in the raw and wine-making process.

Comparison of the effectiveness of thiamethoxam and its main metabolite clothianidin after foliar spraying and root irrigation to control Myzus persicae on peach.

The green peach aphid, Myzus persicae, is one of the most economically important pests in peach-growing areas around the world. In many countries, the application of insecticides is the main method to control and reduce the population of M. persicae. In this study, we investigated the effects and persistence of thiamethoxam against M. persicae by foliar spraying and root irrigation. The residues of thiamethoxam and clothianidin in peach were determined to assess food safety. The results showed that thiamethoxam treatment significantly reduced the population of M. persicae through foliar spraying and root irrigation. And the persistence of root irrigation on M. persicae was significantly longer than that of spraying. Thiamethoxam and clothianidin were absorbed by the roots, transported to other parts of the plant, and concentrated in the leaves, especially new leaves. The final residues of thiamethoxam and clothianidin in peaches were below the maximum residue limit (MRLs). These results suggested that thiamethoxam is more effective in M. persicae control through root irrigation than foliar spraying. The persistence of root irrigation on M. persicae was significantly longer than that of spraying. These results shed some light upon the control of M. persicae by root irrigation of thiamethoxam.

Simultaneous determination of spirodiclofen, spiromesifen, and spirotetramat and their relevant metabolites in edible fungi using ultra-performance liquid chromatography/tandem mass spectrometry.

A fast, sensitive, and reliable analytical method was developed and validated for simultaneous identification and quantification of spirodiclofen, spiromesifen, and spirotetramat and their relevant metabolites in edible fungi by ultra-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS). First, sample extraction was done with acetonitrile containing 1% formic acid followed by phase separation with the addition of MgSO4:NaOAc. Then, the supernatant was purified by primary secondary amine (PSA), octadecylsilane (C18), and graphitized carbon black (GCB). The linearities of the calibrations for all analytes were excellent (R2 ≥ 0.9953). Acceptable recoveries (74.5-106.4%) for all analytes were obtained with good intra- and inter- relative standard deviations of less than 14.5%. The limit of quantification (LOQs) for all analytes was 10 µg kg-1. For accurate quantification, matrix-matched calibration curve was applied to normalize the matrix effect. The results indicated that the method was suitable for detecting the three acaricides and their relevant metabolites in edible fungi.

Development of a fast multi-residue method for the determination of succinate dehydrogenase inhibitor fungicides in cereals, vegetables and fruits by modified QuEChERS and UHPLC-MS/MS.

In this study, a simple, sensitive and efficient analytical method was developed and validated for the determination of 11 fungicides in grape, potato, rice, corn, wheat and soybean using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The fungicides were extracted using acetonitrile, and then the extracts were cleaned up using primary secondary amine (PSA), octadecylsilane (C18) and graphitized carbon black (GCB). Method validation displayed acceptable linearities for all pesticides (R2 greater than 0.99). The calculated limits of detection and quantification were 1 µg kg-1 and 10 µg kg-1 in different matrices, respectively. It was necessary to employ matrix-matched standards for correct quantification of these fungicides in all matrices. Acceptable recoveries (75.7-104.2%) were obtained for all the target compounds with good intra-day and inter-day precisions (RSD ≤ 11.9%). The results indicated that the method is reliable for detecting these fungicides in various foods.

Method development and validation of ten pyrethroid insecticides in edible mushrooms by Modified QuEChERS and gas chromatography-tandem mass spectrometry.

A method for simultaneous determination of ten pyrethroid insecticides residues in edible mushrooms was developed. The samples were pretreated by a quick, easy, cheap, effective, rugged (QuEChERS) method. The ten pyrethroid insecticides were extracted from six kinds of edible mushrooms using acetonitrile and subsequently cleaned up by octadecylsilane (C18) or primary secondary amine (PSA). Instrumental analysis was completed in 16 min using gas chromatography-tandem mass spectrometry (GC-MS/MS). The overall average recoveries in the six kinds of edible mushrooms at three levels (10, 100 and 1000 µg kg-1) ranged from 72.8% to 103.6%. The intraday and interday relative standard deviations (RSD) were lower than 13.0%. The quantification limits were below 5.57 µg kg-1 in different matrices. The results demonstrated that the method is convenient for the quick detection of pyrethroid insecticides in edible mushrooms.

MicroRNA858 negatively regulates anthocyanin biosynthesis by repressing AaMYBC1 expression in kiwifruit (Actinidia arguta).

The anthocyanin biosynthetic pathway regulated by exogenous and endogenous factors through sophisticated networks has been extensively studied in kiwifruit (Actinidia arguta). However, the role of micro RNAs (miRNAs) as regulatory factor in this process is largely unclear. Here, we demonstrate that miR858 is a negative regulator of anthocyanin biosynthesis by repressing the target gene AaMYBC1 in red-colored kiwifruit. Transient co-transformation in Nicotiana benthamiana confirmed that miR858 could target AaMYBC1, which was identified to be an R2R3-type tanscription factor (TF). Subcellular localization showed that AaMYBC1 was located in the nucleus, indicating AaMYBC1 protein could act as a transcriptional regulator in plant cells. Functional protein association network analysis and the yeast two hybrid (Y2H) assay revealed that AaMYBC1 and AabHLH42 interact with each other. Silencing of AaMYBC1 using the virus-induced gene silencing method in the core of A. arguta 'HB' ('Hongbaoshixing', a kind of red-fleshed A. arguta cultivar) fruits reduced the accumulation of anthocyanin and decreased the expression of late biosynthetic genes. miR858 overexpression played a stronger role than AaMYBC1 silencing in the inhibition of coloration. With overexpression of miR858, A. arguta did not present coloration, and anthocyanin was hardly detected. Together, these results clarify the negative regulatory role of miR858 in mediating anthocyanin biosynthesis and accumulation in A. arguta, providing novel insights into the molecular mechanism of anthocyanin biosynthesis.