Residue Degradation and Risk Assessment of Difenoconazole and Its Metabolite during Tea Growing, Processing and Brewing by Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry Determination.

Residue dissipation and risk assessment of difenoconazole and its metabolite difenoconazole-alcohol during tea growing, processing, and brewing was first investigated by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The limits of quantification for both difenoconazole and difenoconazole-alcohol were 0.001 mg/kg in fresh tea leaves and tea, and 0.0002 mg/L in tea infusion. In field trials, the dissipation half-lives of difenoconazole in fresh tea leaves was 1.77 days. After spraying, the residues of difenoconazole-alcohol increased and then gradually dissipated like difenoconazole. After 14 days, the dissipation rates of difenoconazole and difenoconazole-alcohol reached 99%. When fresh tea leaves were harvested on different days, the total processing factors (PFs) of difenoconazole and difenoconazole-alcohol for green tea were 0.86-1.05 and 0.78-0.85, respectively, while the total PFs for black tea were 0.83-1.13 and 0.82-1.66, respectively. Metabolism of difenoconazole was accelerated during tea processing. When brewing black tea, the leaching rates (LRs) of difenoconazole and difenoconazole-alcohol were 8.4-17.9% and 31.8-38.9%, respectively, while when brewing green tea, the LRs were 15.4-23.5% and 30.4-50.6%, respectively. The LRs of difenoconazole and difenoconazole-alcohol in black tea were higher than those in green tea. The potential threat to human health for dietary intake of difenoconazole and difenoconazole-alcohol residues from tea consumption is negligible. However, the dietary risk of difenoconazole in fruits and vegetables that are essential for daily diets is concerning, with a risk probability of 158%.

Mitogen-activated protein kinase MxMPK3-2 mediated phosphorylation of MxZR3.1 participates in regulating iron homoeostasis in apple rootstocks.

The micronutrient iron plays a crucial role in the growth and development of plants, necessitating meticulous regulation for its absorption by plants. Prior research has demonstrated that the transcription factor MxZR3.1 restricts iron absorption in apple rootstocks; however, the precise mechanism by which MxZR3.1 contributes to the regulation of iron homoeostasis in apple rootstocks remains unexplored. Here, MxMPK3-2, a protein kinase, was discovered to interact with MxZR3.1. Y2H, bimolecular fluorescence complementation and pull down experiments were used to confirm the interaction. Phosphorylation and cell semi-degradation tests have shown that MxZR3.1 can be used as a substrate of MxMPK3-2, which leads to the MxZR3.1 protein being more stable. In addition, through tobacco transient transformation (LUC and GUS) experiments, it was confirmed that MxZR3.1 significantly inhibited the activity of the MxHA2 promoter, while MxMPK3-2 mediated phosphorylation at the Ser94 site of MxZR3.1 further inhibited the activity of the MxHA2 promoter. It is tightly controlled to absorb iron during normal growth and development of apple rootstocks due to the regulatory effect of the MxMPK3-2-MxZR3.1 module on MxHA2 transcription level. Consequently, this research has revealed the molecular basis of how the MxMPK3-2-MxZR3.1 module in apple rootstocks controls iron homoeostasis by regulating the MxHA2 promoter's activity.

miR164-MhNAC1 regulates apple root nitrogen uptake under low nitrogen stress.

Nitrogen is an essential nutrient for plant growth and serves as a signaling molecule to regulate gene expression inducing physiological, growth and developmental responses. An excess or deficiency of nitrogen may have adverse effects on plants. Studying nitrogen uptake will help us understand the molecular mechanisms of utilization for targeted molecular breeding. Here, we identified and functionally validated an NAC (NAM-ATAF1/2-CUC2) transcription factor based on the transcriptomes of two apple rootstocks with different nitrogen uptake efficiency. NAC1, a target gene of miR164, directly regulates the expression of the high-affinity nitrate transporter (MhNRT2.4) and citric acid transporter (MhMATE), affecting root nitrogen uptake. To examine the role of MhNAC1 in nitrogen uptake, we produced transgenic lines that overexpressed or silenced MhNAC1. Silencing MhNAC1 promoted nitrogen uptake and citric acid secretion in roots, and enhanced plant tolerance to low nitrogen conditions, while overexpression of MhNAC1 or silencing miR164 had the opposite effect. This study not only revealed the role of the miR164-MhNAC1 module in nitrogen uptake in apple rootstocks but also confirmed that citric acid secretion in roots affected nitrogen uptake, which provides a research basis for efficient nitrogen utilization and molecular breeding in apple.

Genetic variations in MdSAUR36 participate in the negative regulation of mesocarp cell division and fruit size in Malus species.

Final fruit size of apple (Malus domestica) cultivars is related to both mesocarp cell division and cell expansion during fruit growth, but it is unclear whether the cell division and/or cell enlargement determine most of the differences in fruit size between Malus species. In this study, by using an interspecific hybrid population between Malus asiatica "Zisai Pearl" and Malus domestica cultivar "Red Fuji," we found that the mesocarp cell number was the main causal factor of diversity in fruit size between Malus species. Rapid increase in mesocarp cell number occurred prior to 28 days after anthesis (DAA), while cell size increased gradually after 28 DAA until fruit ripening. Six candidate genes related to auxin signaling or cell cycle were predicted by combining the RNA-seq data and previous QTL data for fruit weight. Two InDels and 10 SNPs in the promoter of a small auxin upregulated RNA gene MdSAUR36 in Zisai Pearl led to a lower promoter activity than that of Red Fuji. One non-synonymous SNP G/T at 379 bp downstream of the ATG codon of MdSAUR36, which was heterozygous in Zisai Pearl, exerted significant genotype effects on fruit weight, length, and width. Transgenic apple calli by over-expressing or RNAi MdSAUR36 confirmed that MdSAUR36 participated in the negative regulation of mesocarp cell division and thus apple fruit size. These results could provide new insights in the molecular mechanism of small fruit size in Malus accession and be potentially used in molecular assisted breeding via interspecific hybridization. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01441-4.

Review of published 467 achondroplasia patients: clinical and mutational spectrum.

AIM: Achondroplasia is the most common of the skeletal dysplasias that cause fatal and disabling growth and developmental disorders in children, and is caused by a mutation in the fibroblast growth factor receptor, type 3 gene(FGFR3). This study aims to analyse the clinical characteristics and gene mutations of ACH to accurately determine whether a patient has ACH and to raise public awareness of the disease. METHODS: The database of Pubmed, Cochrane Library, Wanfang and CNKI were searched with terms of "Achondroplasias" or "Skeleton-Skin-Brain Syndrome" or "Skeleton Skin Brain Syndrome" or "ACH" and "Receptor, Fibroblast Growth Factor, Type 3" or "FGFR3". RESULTS: Finally, four hundred and sixty-seven patients with different FGFR3 mutations were enrolled. Of the 138 patients with available gender information, 55(55/138, 40%) were female and 83(83/138, 60%) were male. Among the patients with available family history, 47(47/385, 12%) had a family history and 338(338/385, 88%) patients were sporadic. The age of the patients ranged from newborn babies to 36 years old. The mean age of their fathers was 37 ± 7 years (range 31-53 years). Patients came from 12 countries and 2 continents, with the majority being Asian (383/432, 89%), followed by European (49/432, 11%). Short stature with shortened arms and legs was found in 112(112/112) patients, the abnormalities of macrocephaly in 94(94/112) patients, frontal bossing in 89(89/112) patients, genu valgum in 64(64/112) patients and trident hand were found in 51(51/112) patients. The most common mutation was p.Gly380Arg of the FGFR3 gene, which contained two different base changes, c.1138G > A and c.1138G > C. Ten rare pathogenic mutations were found, including c.831A > C, c.1031C > G, c.1043C > G, c.375G > T, c.1133A > G, c.1130T > G, c.833A > G, c.649A > T, c.1180A > T and c.970_971insTCTCCT. CONCLUSION: ACH was caused by FGFR3 gene mutation, and c.1138G > A was the most common mutation type. This study demonstrates the feasibility of molecular genetic testing for the early detection of ACH in adolescents with short stature, trident hand, frontal bossing, macrocephaly and genu valgum.

Pan-genome analysis of 13 Malus accessions reveals structural and sequence variations associated with fruit traits.

Structural variations (SVs) and copy number variations (CNVs) contribute to trait variations in fleshy-fruited species. Here, we assemble 10 genomes of genetically diverse Malus accessions, including the ever-green cultivar 'Granny Smith' and the widely cultivated cultivar 'Red Fuji'. Combining with three previously reported genomes, we assemble the pan-genome of Malus species and identify 20,220 CNVs and 317,393 SVs. We also observe CNVs that are positively correlated with expression levels of the genes they are associated with. Furthermore, we show that the noncoding RNA generated from a 209 bp insertion in the intron of mitogen-activated protein kinase homology encoding gene, MMK2, regulates the gene expression and affects fruit coloration. Moreover, we identify overlapping SVs associated with fruit quality and biotic resistance. This pan-genome uncovers possible contributions of CNVs to gene expression and highlights the role of SVs in apple domestication and economically important traits.

Active Targeted Janus Theranostic Nanoplatforms Enable Chemo-Photothermal Therapy to Inhibit the Growth of Breast Cancer.

Nanoscale structures have been developed to serve various functions in cancer therapy, encompassing areas such as diagnosis, biomedical visualization, tissue regeneration, and drug delivery. Based on biocompatible chitosan oligosaccharides (COS) and gold nanorods (GNRs), we designed the drug delivery systems (GNR@polyacrylic acid-Mn@COS Janus nanoparticles (JNPs)), which achieved paclitaxel (PTX) loaded on the side of GNRs, and the PAA-Mn domain served as magnetic resonance imaging contrast agents. This system was found to be effectively delivered to tumor sites through the enhanced permeability and retention (EPR) effect and the active target of the COS. The uniform JNPs selectively targeted cancer cells instead of normal cells through interacting with the COS on the surface of tumor cells, and the pH/NIR-responsive drug release behavior further enhanced their therapeutic effects. The in vivo effects of JNPs against tumors were evaluated using subcutaneous and orthotopic lung metastasis models, yielding promising outcomes for both tumor diagnosis and cancer treatment. In conclusion, the obtained JNPs hold great promise as a theranostic nanoplatform with synergistic chemotherapeutic and photothermal effects.

A Turn-On Fluorescence Sensor Based on Nitrogen-Doped Carbon Dots and Cu2+ for Sensitively and Selectively Sensing Glyphosate.

Glyphosate has excellent herbicidal activity, and its extensive use may induce residue in the environment and enter into humans living through the food chain, causing negative impact. Here, water-soluble 1.55 nm size nitrogen-doped carbon quantum dots (NCDs) with strong blue fluorescence were synthesized using sodium citrate and adenine. The maximum excitation and emission wavelengths of NCDs were 380 nm and 440 nm, respectively. The above synthesized NCDs were first used for the construction of a fluorescence sensor for glyphosate detection. It was found that Cu2+ could quench the fluorescence of NCDs effectively through the photoinduced electron transfer (PET) process, which was confirmed using fluorescence lifetime measurements. Additionally, the fluorescence was restored with the addition of glyphosate. Hence, a sensitive turn-on fluorescence sensor based on NCDs/Cu2+ for glyphosate analysis was developed. The LODs of glyphosate for water and rice samples were recorded as 0.021 µg/mL and 0.049 µg/mL, respectively. The sensor was applied successfully for ultrasensitive and selective detection of glyphosate in environmental water and rice samples with satisfied recoveries from 82.1% to 113.0% using a simple sample pretreatment technique. The proposed strategy can provide a significant potential for monitoring glyphosate residue in water and agricultural product samples.

MdGRF11-MdARF19-2 module acts as a positive regulator of drought resistance in apple rootstock.

14-3-3 proteins play an important role in the response of plants to drought resistance. In this study, 14-3-3 protein MdGRF11 was cloned from Malus xiaojinensis, and its positive regulation of drought resistance was verified using Orin calli and M. xiaojinensis plants. The transcription factor MdARF19-2 was further screened for interaction with this protein in vitro and in vivo. We also conducted experiments using Orin calli and found that the overexpression of MdARF19-2 decreased the level of reactive oxygen species (ROS) and increased the activity of enzymes that scavenge ROS in plant materials. This indicates that MdARF19-2 is a positive regulator in the drought resistance of plants. The drought tolerance was further improved by the overexpression of both MdGRF11 and MdARF19-2 in the calli. In addition, we examined several genes related to ROS scavenging with auxin response factor binding elements in their promoters and found that their level of expression was regulated by the MdGRF11-MdARF19-2 module. In conclusion, the enhancement of plant drought resistance by MdGRF11 could be owing to its accumulation at the protein level in response to drought, which then combined with MdARF19-2, affecting the expression of MdARF19-2 downstream genes. Thus, it scavenges ROS, which ultimately improves the resistance of plant to drought stress.

Synergistic effects of multitype carbon doping and oxygen vacancies in TiO2/CNTs composite fabricated via nonthermal plasma for formaldehyde removal.

Formaldehyde as one of the typical indoor pollutants has long been concerned as it can pose a threat to human health. TiO2/CNTs composite with oxygen vacancies and multitype carbon doping (C-TiO2/CNTs) was fabricated using nonthermal plasma for the photocatalytic degradation of formaldehyde. The maximum degradation rate of formaldehyde was 93% and 83% via the new catalyst (with 5% CNTs content) under solar and visible light, respectively. The characterization of the catalyst confirmed the in-situ multitype carbon doping and oxygen vacancies: interstitial carbon doping and oxygen vacancies could dramatically reduce the bandgap and contribute to the improved absorption capability of formaldehyde and electrons. Interfacial carbon doping in the form of C-O-Ti bonds provided a migration channel, whereby photogenerated electrons could efficiently transfer from CNTs to TiO2 and then quench the holes left in the VB of TiO2. Therefore, the multitype carbon doping and oxygen vacancies can expand the light response as well as promote the separation of photo-generated electron/hole pairs. EPR results and experiment section indicated that O2·- plays the most significant role in formaldehyde removal due to the reverse transfer of the electrons. This work advances the understanding of photo-degradation of TiO2/CNTs composite and provides a new route for the abatement of formaldehyde.

Calcium Orthophosphate in Liposomes for Co-Delivery of Doxorubicin Hydrochloride/Paclitaxel in Breast Cancer.

Nanoparticles (NPs) show great advantages in cancer treatment by enabling controlled and targeted delivery of payloads to tumor sites through the enhanced permeability and retention (EPR) effect. In this study, highly effective pH-responsive and biodegradable calcium orthophosphate@liposomes (CaP@Lip) NPs with a diameter of 110 ± 20 nm were designed and fabricated. CaP@Lip NPs loaded with hydrophobic paclitaxel and hydrophilic doxorubicin hydrochloride achieved excellent drug loading efficiencies of 70 and 90%, respectively. Under physiological conditions, the obtained NPs are negatively charged. However, they switched to positively charged when exposed to weak acidic environments by which internalization can be promoted. Furthermore, the CaP@Lip NPs exhibit an obvious structural collapse under acid conditions (pH 5.5), which confirms their excellent biodegradability. The "proton expansion" effect in endosomes and the pH-responsiveness of the NPs facilitate the release of encapsulated drugs from individual channels. The effectiveness and safety of the drug delivery systems were demonstrated through in vitro and in vivo experiments, with a 76% inhibition of tumor growth. These findings highlight the high targeting ability of the drug-loaded NPs to tumor sites through the EPR effect, effectively suppressing tumor growth and metastasis. By combining CaP NPs and liposomes, this study not only resolves the toxicity of CaP but also enhances the stability of liposomes. The CaP@Lip NPs developed in this study have significant implications for biomedical applications and inspire the development of intelligent and smart drug nanocarriers and release systems for clinical use.

MxMPK6-2-mediated phosphorylation enhances the response of apple rootstocks to Fe deficiency by activating PM H+ -ATPase MxHA2.

Iron (Fe) deficiency significantly affects the growth and development, fruit yield and quality of apples. Apple roots respond to Fe deficiency stress by promoting H+ secretion, which acidifies the soil. In this study, the plasma membrane (PM) H+ -ATPase MxHA2 promoted H+ secretion and root acidification of apple rootstocks under Fe deficiency stress. H+ -ATPase MxHA2 is upregulated in Fe-efficient apple rootstock of Malus xiaojinensis at the transcription level. Fe deficiency also induced kinase MxMPK6-2, a positive regulator in Fe absorption that can interact with MxHA2. However, the mechanism involving these two factors under Fe deficiency stress is unclear. MxMPK6-2 overexpression in apple roots positively regulated PM H+ -ATPase activity, thus enhancing root acidification under Fe deficiency stress. Moreover, co-expression of MxMPK6-2 and MxHA2 in apple rootstocks further enhanced PM H+ -ATPase activity under Fe deficiency. MxMPK6-2 phosphorylated MxHA2 at the Ser909 site of C terminus, Thr320 and Thr412 sites of the Central loop region. Phosphorylation at the Ser909 and Thr320 promoted PM H+ -ATPase activity, while phosphorylation at Thr412 inhibited PM H+ -ATPase activity. MxMPK6-2 also phosphorylated the Fe deficiency-induced transcription factor MxbHLH104 at the Ser169 site, which then could bind to the promoter of MxHA2, thus enhancing MxHA2 upregulation. In conclusion, the MAP kinase MxMPK6-2-mediated phosphorylation directly and indirectly regulates PM H+ -ATPase MxHA2 activity at the protein post-translation and transcription levels, thus synergistically enhancing root acidification under Fe deficiency stress.

Genome-wide analysis of the MdZR gene family revealed MdZR2.2-induced salt and drought stress tolerance in apple rootstock.

The DNL-type zinc finger protein constitutes a zinc ribbon protein (ZR) family, which belongs to a branch of zinc finger protein and plays an essential role in response to abiotic stress. Here, we identified six apple (Malus domestica) MdZR genes. Based on their phylogenetic relationship and gene structure, the MdZR genes were divided into three categories, including MdZR1, MdZR2, and MdZR3. Subcellular results showed that the MdZRs are located on the nuclear and membrane. The transcriptome data showed that MdZR2.2 is expressed in various tissues. The expression analysis results showed that MdZR2.2 was significantly upregulated under salt and drought treatments. Thus, we selected MdZR2.2 for further research. Overexpression of MdZR2.2 in apple callus improved their tolerance to drought and salt stress and ability to scavenge reactive oxygen species (ROS). In contrast, transgenic apple roots with silenced MdZR2.2 grew more poorly than the wild type when subjected to salt and drought stress, which reduced their ability to scavenge ROS. To our knowledge, this is the first study to analyze the MdZR protein family. This study identified a gene that responds to drought and salt stress. Our findings lay a foundation for a comprehensive analysis of the MdZR family members.

Method validation for detection of afidopyropen and M440I007 in tea.

BACKGROUND: Afidopyropen is a novel biorational insecticide for controlling piercing pests with great potential for application in tea gardens that can form the metabolite M440I007 when utilized for crops. However, because of a lack of analytical method for afidopyropen and M440I007 in tea, there is no means of monitoring the residues. Therefore, method development, validation and simultaneous determination of afidopyropen and M440I007 in fresh tea leaves, dried tea and tea infusion is of prime significance. RESULTS: A TPT cartridge-based method was developed for the solid phase extraction of afidopyropen and M440I007 from tea matrices. Extraction and clean-up conditions, including the composition, volume and temperature of elutions, were optimized to achieve the best results. Both targets were extracted using water and acetonitrile, with a water:acetonitrile (v/v) ratio of 4:10 for fresh leaves and 8:10 for dried tea, which were then cleaned and analyzed using ultraperformance liquid chromatography-tandem mass spectrometry. Both analytes demonstrated excellent linearity with a correlation coefficient above 0.998. The optimized analytical method offered limits of quantifications of 0.005, 0.005 and 0.002 mg kg-1 (converted to dried tea) in fresh tea shoots, dried tea and tea infusion for both targets, respectively. Average recoveries of afidopyropen and M440I007 ranged from 79.0% to 101.5%, with relative standard deviations ≤ 14.7%. CONCLUSION: The results showed that the method of determination for these insecticides in tea matrices was practical and efficient. © 2023 Society of Chemical Industry.

The relationships between thyroid functions of short-term rapid hypothyroidism and blood lipid levels in post-thyroidectomy patients of differentiated thyroid cancer.

Objective: To explore the relationship between short-term rapid hypothyroidism and blood lipid levels in patients with differentiated thyroid cancer (DTC). Methods: Seventy-five DTC patients scheduled to receive radioactive iodine ablation were enrolled. Levels of thyroid hormone and serum lipids were tested at two time points: the euthyroid before thyroidectomy, and the hypothyroid (off thyroxine). Then the collected data were analyzed. Results: Totally 75 DTC patients enrolled, among them, 5o were female (66.67%) and 25 were male (33. 33%), with an average age of 52.24 ± 1.24 years old. The short-term rapid severe hypothyroidism induced by thyroid hormone withdrawal significantly aggravated dyslipidemia, particularly in patients with dyslipidemia before thyroidectomy (All P < 0.01). However, there was no significant differences between blood lipid levels with different thyroid stimulating hormone (TSH) levels. And our study showed significant negative correlations between free triiodothyronine levels and the changes from euthyjroidism to hypothyroidism in total cholesterol (r=-0.31, P=0.03), triglycerides (r=-0.39, P=0.006), high density lipoprotein-cholesterol (HDL-C) (r=-0.29, P=0.042), and significant positive correlations between free thyroxine and the changes of HDL-C (r=-0.32, P=0.027) were identified in females, however, which were not observed in males. Conclusion: Short-term rapids severe hypothyroidism caused by thyroid hormone withdrawal can lead to rapid significant changes in blood lipid levels. It is necessary to pay attention to dyslipidemia and its long-term effects after thyroid hormone withdrawal, especially in patients with dyslipidemia before thyroidectomy. Clinical trial registration: https://clinicaltrials.gov/ct2/show/NCT03006289?term=NCT03006289&draw=2&rank=1, identifier NCT03006289.

Association of polymorphisms in Th1/Th2-related cytokines (IFN-γ, TGFß1, IL-1ß, IL-2, IL-4, IL-18) with oral lichen planus: A pooled analysis of case-control studies.

Background/purpose: Increasing evidence suggests that single-nucleotide polymorphisms (SNPs) in Th1/Th2-related cytokine genes correlated with oral lichen planus (OLP) susceptibility. However, these results were inconsistent and inconclusive. Hence, the aim of this study is to draw a more precise estimation of the genetic associations between SNPs in 6 cytokines (IFN-γ, IL-18, TGFß1, IL-1ß, IL-2, IL-4) and OLP. Materials and methods: A systematic literature search was conducted to identify all eligible case-control studies on the association between SNPs in 6 cytokines and OLP susceptibility. Odds ratios (ORs) and 95% confidence intervals (CIs) from each study were pooled to estimate the strength of the association. Results: A significant association of IFN-γ (874A/T) polymorphism with OLP was found (OR, 1.49; 95%CI, 1.22-1.81; P < 0.001) based on 6 eligible studies. A significant association of IL-18 (137G/C) polymorphism with OLP was found (OR, 1.64; 95%CI, 1.24-2.18; P < 0.001) based on 3 studies. A marginally significant association of TGFß1 (509C/T) polymorphism in allele model with OLP was found (OR, 1.31; 95%CI, 1.01-1.71; P = 0.05) based on 4 studies. Nevertheless, lack of significant association of IL-1ß (3954C/T), IL-2 (330T/G), IL-4 (590C/T), and IL-18 (607C/A) polymorphisms with OLP was found (P > 0.05) based on 3 studies, respectively. Conclusion: This is the first meta-analysis to investigate the associations of 6 cytokines polymorphisms with OLP, suggesting that SNPs in IFN-γ, IL-18, and TGFß1 may act as genetic factors for OLP risk. Further well-designed studies with larger sample size and multiple ethnicities are needed to validate these associations.

SPE-UPLC-MS/MS for Determination of 36 Monomers of Alkylphenol Ethoxylates in Tea.

Alkylphenol ethoxylates (APEOs) represent a non-ionic surfactant widely used as adjuvants in pesticide formulation, which is considered to cause an endocrine-disrupting effect. In the current study, we established a detection method for the APEOs residue in tea based on solid-phase extraction (SPE) for the simultaneous analysis of nonylphenol ethoxylates (NPEOs) and octylphenol ethoxylates (OPEOs) by UPLC-MS/MS. In the spiked concentrations from 0.024 to 125.38 µg/kg for 36 monomers of APEOs (nEO = 3-20), the recoveries of APEOs range from 70.3-110.7% with RSD ≤ 16.9%, except for OPEO20 (61.8%) and NPEO20 (62.9%). The LOQs of OPEOs and NPEOs are 0.024-6.27 and 0.16-5.01 µg/kg, respectively. OPEOs and NPEOs are detected in 50 marketed tea samples with a total concentration of 0.057-12.94 and 0.30-215.89 µg/kg, respectively. The detection rate and the range of the monomers of NPEOs are generally higher than those of OPEOs. The current study provides a theoretical basis for the rational use of APEOs as adjuvants in commercial pesticide production.

The metabolism and dissipation behavior of tolfenpyrad in tea: A comprehensive risk assessment from field to cup.

The metabolites of pesticides usually require rational risk assessment. In the present study, the metabolites of tolfenpyrad (TFP) in tea plants were identified using UPLC-QToF/MS analysis, and the transfer of TFP and its metabolites from tea bushes to consumption was studied for a comprehensive risk assessment. Four metabolites, PT-CA, PT-OH, OH-T-CA, and CA-T-CA, were identified, and PT-CA and PT-OH were detected along with dissipation of the parent TFP under field conditions. During processing, 3.11-50.00 % of TFP was further eliminated. Both PT-CA and PT-OH presented a downward trend (7.97-57.89 %) during green tea processing but an upward trend (34.48-124.17 %) during black tea manufacturing. The leaching rate (LR) of PT-CA (63.04-101.03 %) from dry tea to infusion was much higher than that of TFP (3.06-6.14 %). As PT-OH was no longer detected in tea infusions after 1 d of TFP application, TFP and PT-CA were taken into account in the comprehensive risk assessment. The risk quotient (RQ) assessment indicated a negligible health risk, but PT-CA posed a greater potential risk than TFP to tea consumers. Therefore, this study provides guidance for rational TFP application and suggests the sum of TFP and PT-CA residues as the maximum residual limit (MRL) in tea.

MdbZIP74 negatively regulates osmotic tolerance and adaptability to moderate drought conditions of apple plants.

Drought is the most prominent threat to global agricultural production. The basic leucine zipper (bZIP) family is related to the response to a series of abiotic stress. In this case, apple calli and the seedlings of MdbZIP74-RNAi transgenic lines were obtained. Under osmotic stress and moderate drought conditions, the content of malondialdehyde, relative water content and other stress-related assays were measured. MdbZIP74 was found to negatively regulate the osmotic tolerance of apple callus. The growth of MdbZIP74-RNAi calli enhanced resistance without significant production loss. The silencing of MdbZIP74 contributes to redox balance and the adaptability of apple seedlings to moderate drought conditions. Four related differentially expressed genes in the biosynthesis of cytokinin and catabolic pathway were identified through a transcriptome analysis of MdbZIP74-RNAi seedlings under moderate drought conditions. MdLOG8 was further identified as the target of MdbZIP74 involved in the drought adaptability of apple plants using a dual experiment. Further confirmation showed MdLOG8 was maintained in the MdbZIP74-RNAi seedlings presumably acting as the growth regulator to enhance drought adaptability. It was concluded that the correct regulation of cytokinin level under moderate drought conditions maintains the redox balance and avoids the situation of plants surviving with the minimal resources.

Method Validation for Multi-Pesticide Residue Determination in Chrysanthemum.

The chrysanthemum can be consumed in various forms, representing the "integration of medicine and food". Quantitative analysis of multi-pesticide residues in chrysanthemum matrices is therefore crucial for both product-safety assurance and consumer-risk evaluation. In the present study, a simple and effective method was developed for simultaneously detecting 15 pesticides frequently used in chrysanthemum cultivation in three matrices, including fresh flowers, dry chrysanthemum tea, and infusions. The calibration curves for the pesticides were linear in the 0.01-1 mg kg-1 range, with correlation coefficients greater than 0.99. The limits of quantification (LOQs) for fresh flowers, dry chrysanthemum tea, and infusions were 0.01-0.05 mg kg-1, 0.05 mg kg-1, and 0.001-0.005 mg L-1, respectively. In all selected matrices, satisfactory accuracy and precision were achieved, with recoveries ranging from 75.7 to 118.2% and relative standard deviations (RSDs) less than 20%. The validated method was then used to routinely monitor pesticide residues in 50 commercial chrysanthemum-tea samples. As a result, 56% of samples were detected with 5-13 pesticides. This research presents a method for the efficient analysis of multi-pesticide residues in chrysanthemum matrices.