The Cys-2088-Arg mutation in the ACCase gene and enhanced metabolism confer cyhalofop-butyl resistance in Chinese sprangletop (Leptochloa chinensis).

Acetyl-CoA carboxylase (ACCase)-inhibiting herbicides are among the most commonly used herbicides to control grassy weeds, especially Leptochloa chinensis, in rice fields across China. Herein, we collected a suspected resistant (R) population of L. chinensis (HFLJ16) from Lujiang county in Anhui Province. Whole plant dose response tests showed that, compared with the susceptible (S) population, the R population showed high resistance to cyhalofop-butyl (22-fold) and displayed cross-resistance to metamifop (9.7-fold), fenoxaprop-P-ethyl (18.7-fold), quizalofop-P-ethyl (7.6-fold), clodinafop-propargyl (12-fold) and clethodim (8.4-fold). We detected an amino acid substitution (Cys-2088-Arg) in the ACCase of resistant L. chinensis. However, ACCase gene expression levels were not significantly different (P > 0.05) between R plants and S plants, without or with cyhalofop-butyl treatment. Furthermore, pretreatment with piperonyl butoxide (PBO, a cytochrome P450 monooxygenase (CYP450) inhibitor) or 4-chloro-7-nitrobenzoxadiazole (NBD-Cl, a glutathione-S-transferase (GST) inhibitor), inhibited the resistance of the R population to cyhalofop-butyl significantly (by approximately 60% and 26%, respectively). Liquid chromatography tandem mass spectrometry analysis showed that R plants metabolized cyhalofop-butyl and cyhalofop acid (its metabolite) significantly faster than S plants. Three CYP450 genes, one GST gene, and two ABC transporter genes were induced by cyhalofop-butyl and were overexpressed in the R population. Overall, GST-associated detoxification, CYP450 enhancement, and target-site gene mutation are responsible for the resistance of L. chinensis to cyhalofop-butyl.

Effects of different fertilization practices on maize yield, soil nutrients, soil moisture, and water use efficiency in northern China based on a meta-analysis.

The application of fertilizer to ensure the steady improvement of crop yield has become the main means of agricultural production. However, it remains to be determined whether fertilization practices with different combinations of nitrogen (N), phosphorus (P), potassium (K), and organic (O) fertilizers play a positive role in the sustainability of maize yield and the soil in which it is grown. Therefore,this meta-analysis extracted 2663 data points from 76 studies to systematically analyze and explore the effects of different fertilization measures on maize yield, soil nutrients, water content and water use efficiency (WUE) in northern China. Articles addressing this topic showed that fertilization effectively increased the soil nutrient content and maize yield. The soil organic matter (SOM) increased by 2.36 (N)-55.38% (NPO), total nitrogen content increased by 6.10 (N)-56.39% (NPO), available phosphorus content increased by 17.12 (N)-474.74% (NPO), and available potassium content changed by - 2.90 (NP)-64.40% (NPO). Soil moisture increased by 3.59% under a single organic fertilizer application and decreased by 4.27-13.40% under the other treatments. Compared with no fertilization, the yield increase of fertilized maize reached 11.65-220.42%. NP, NPK and NPKO contributed the most to increased yield in lithological, black and fluvo-aquic soils, respectively. The effects of different fertilization practices on maize yield varied in response to the same meteorological factors. The WUE increased from 9.51 to 160.72%. In conclusion, rational fertilization can improve the soil nutrient content and increase maize yield. The combined application of chemical and organic fertilizer showed the greatest increase in yield and WUE. Organic fertilizer application alone increased soil moisture. Our results provide a theoretical basis for fertilizer application and for improving the soil structure for maize cultivation in northern China.

Transcription factor 12-mediated self-feedback regulatory mechanism is required in DUX4 fusion leukaemia.

BACKGROUND: IGH::DUX4 is frequently observed in 4% B-cell acute lymphoblastic leukaemia patients. Regarding the IGH::DUX4-driven transactivation and alternative splicing, which are the main reasons behind this acute leukaemia outbreak, it remains unclear how transcriptional cofactors contribute to this oncogenic process. Further investigation is required to elucidate their specific role in leukaemogenesis. METHODS: In order to investigate the cofactors of IGH::DUX4, integrated mining of Chromatin immunoprecipitation (ChIP)-sequencing and RNA-sequencing of leukaemia cells and patient samples were conducted. Furthermore, to elucidate the synergistic interaction between transcription factor 12 (TCF12) and IGH::DUX4, knockdown and knockout experiment, mammalian two-hybridisation assay, co-immunoprecipitation and in situ proximity ligation assays were carried out. Additionally, to further investigate the direct interaction between TCF12 and IGH::DUX4, AI-based structural simulations were utilised. Finally, to validate the synergistic role of TCF12 in promoting IGH::DUX4 leukaemia, cell proliferation, apoptosis and drug sensitivity experiments were performed. RESULTS: In this study, we observed that the IGH::DUX4 target gene TCF12 might be an important cofactor/helper for this oncogenic driver. The co-expression of IGH::DUX4 and TCF12 resulted in enhanced DUX4-driven transactivation. Supportively, knockdown and knockout of TCF12 significantly reduced expression of IGH::DUX4-driven target genes in leukaemia REH (a precursor B-cell leukaemia cell line) and NALM-6 cells (a precursor B-cell leukaemia cell line). Consistently, in TCF12 knockout cells, the expression of structure-based TCF12 mutant, but not wild-type TCF12, failed to restore the TCF12-IGH::DUX4 crosstalk and the synergistic transactivation. More importantly, the breakdown in TCF12-IGH::DUX4 cooperation impaired IGH::DUX4-driven leukaemia cell survival, caused sensitivity to the chemotherapy. CONCLUSIONS: Altogether, these results helped to define a previously unrecognised TCF12-mediated positive self-feedback regulatory mechanism in IGH::DUX4 leukaemia, which holds the potential to function as a pivotal drug target for the management of this particular form of leukaemia. HIGHLIGHTS: Transcription factor 12 (TCF12) is a new novel cofactor in IGH::DUX4 transcriptional complexes/machinery. TCF12 mediates a positive self-feedback regulatory mechanism in IGH::DUX4-driven oncogenic transaction. IGH::DUX4-TCF12 structure/cooperation might represent a potent target/direction in future drug design against B-cell acute lymphoblastic leukaemia.

Quantum storage of entangled photons at telecom wavelengths in a crystal.

Quantum storage and distribution of entanglement are the key ingredients for realizing a global quantum internet. Compatible with existing fiber networks, telecom-wavelength entangled photons and corresponding quantum memories are of central interest. Recently, 167Er3+ ions have been identified as a promising candidate for an efficient telecom quantum memory. However, to date, no storage of entangled photons, the crucial step of quantum memory using these promising ions, 167Er3+, has been reported. Here, we demonstrate the storage and retrieval of the entangled state of two telecom photons generated from an integrated photonic chip. Combining the natural narrow linewidth of the entangled photons and long storage time of 167Er3+ ions, we achieve storage time of 1.936 µs, more than 387 times longer than in previous works. Successful storage of entanglement in the crystal is certified using entanglement witness measurements. These results pave the way for realizing quantum networks based on solid-state devices.

Cytochrome P450s-Involved Enhanced Metabolism Contributes to the High Level of Nicosulfuron Resistance in Digitaria sanguinalis from China.

Large crabgrass (Digitaria sanguinalis (L.) Scop.) is one of the major malignant grass weeds in Chinese maize (Zea mays L.) fields, and it has recently developed resistance to the acetolactate synthase (ALS)-inhibiting herbicide nicosulfuron. This study focused on a suspected nicosulfuron-resistant (R) population (LJ-01) of D. sanguinalis, collected from Lujiang County in Anhui Province, China, to explore the resistance level and potential resistance mechanism. Whole-plant dose-response testing confirmed that the LJ-01 population evolved a high level of resistance to nicosulfuron (11.5-fold) compared to the susceptible (S) population, DY-02. The ALS gene sequencing and relative expression assay of the R plants indicated that target gene mutation and overexpression were not responsible for the resistance phenotype. However, pretreatment with malathion, a known cytochrome P450 monooxygenase (P450) inhibitor, alleviated the resistance of the R population to nicosulfuron by approximately 36%. High-performance liquid chromatography (HPLC) analysis revealed that the R plants metabolized nicosulfuron faster than the S plants. Moreover, cross-resistance testing suggested that the R population exhibited low levels of resistance to thifensulfuron-methyl and pyrazosulfuron-ethyl, but it remained susceptible to rimsulfuron. Multiple resistance patterns showed that the R population evolved low resistance to the photosystem inhibitors bromoxynil octanoate and atrazine and sensitivity to the acetyl-CoA carboxylase (ACCase) inhibitor cyhalofop-butyl and the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors tembotrione, mesotrione, and topramezone. This study reports, for the first time, the simultaneous resistance to ALS and different photosystem inhibitors in D. sanguinalis. The nicosulfuron resistance observed in the R population could primarily be attributed to an enhanced metabolism involving P450 enzymes.

HOXD9 is a potential prognostic biomarker involved in immune microenvironment of glioma.

BACKGROUND: Glioma is the prevailing malignant tumor affecting the brain and central nervous system, constituting over 80% of all malignant brain tumors. HOXD9 has been implicated in the development of glioma, but the specific mechanism of its influence on glioma pathogenesis remains incompletely understood. The purpose of this study was to investigate the role of HOXD9 in glioma and examine the changes in HOXD9 expression during the progression of glioma, thus contributing new insights into the pathogenesis of glioma. METHODS: Glioma samples from the Cancer Gene Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) datasets were included in this study. Variations in HOXD9 expression in gliomas between different subgroups of multiple clinical characteristics were explored, and the expression was validated in glioma samples using qRT-PCR and western blotting. Next, the impact of HOXD9 on the prognosis of gliomas was explored by survival analysis, receiver operating characteristic curve, and nomogram plots. Subsequently, the association between HOXD9 and the tumor immune microenvironment was explored using the ssGSEA algorithm and the ESTIMATE algorithm. Then, immune-related pathways associated with HOXD9 were determined by differential express analysis and GSEA. Finally, HOXD9-related genomic alterations were identified. RESULTS: HOXD9 expression is upregulated and correlated with malignant properties in glioma. Similarly, our validation results showed significantly upregulated protein and mRNA levels of HOXD9 in glioma brain tissues. In addition, high HOXD9 expression was indicative of a poor prognosis for glioma patients. Additionally, elevated HOXD9 levels were associated with reduced tumor purity and higher levels of immune invasion. Finally, HOXD9 was significantly associated with genomic alterations. CONCLUSION: Overall, this study has unveiled a significant association between HOXD9 and the prognosis and survival of glioma patients. Our findings highlight the potential of HOXD9 as a prognostic biomarker, implicating its role in influencing the glioma immune microenvironment.

Transcription factor abnormalities in B-ALL leukemogenesis and treatment.

The biological regulation of transcription factors (TFs) and repressor proteins is an important mechanism for maintaining cell homeostasis. In B cell acute lymphoblastic leukemia (B-ALL) TF abnormalities occur at high frequency and are often recognized as the major driving factor in carcinogenesis. We provide an in-depth review of molecular mechanisms of six major TF rearrangements in B-ALL, including DUX4-rearranged (DUX4-R), MEF2D-R, ZNF384-R, ETV6-RUNX1 and TCF3-PBX1 fusions, and KMT2A-R. In addition, the therapeutic options and prognoses for patients who harbor these TF abnormalities are discussed. This review aims to provide an up-to-date panoramic view of how TF-based oncogenic fusions might drive carcinogenesis and impact on potential therapeutic exploration of B-ALL treatments.

Phase separation is required for PML nuclear body biogenesis and function.

PML nuclear body (NB) malfunction often leads to acute leukemia outbreaks and other severe diseases. PML NB rescue is the molecular basis of arsenic success in acute promyelocytic leukemia (APL) treatment. However, it is unclear how PML NBs are assembled. Here, we observed the presence of liquid-liquid phase separation (LLPS) in NB formation by fluorescence recovery after photobleaching (FRAP) experiment. Compared with the wild-type (WT) NBs, PML A216V derived from arsenic-resistant leukemia patients markedly crippled LLPS, but not altered the overall structure and PML RBCC oligomerization. In parallel, we also reported several Leu to Pro mutations that were critical to PML coiled-coil domain. FRAP characterization and comparison between L268P and A216V revealed markedly different LLPS activities in these mutant NBs. Transmission electron microscopy (TEM) inspections of LLPS-crippled and uncrippled NBs showed aggregation- and ring-like PML packing in A216V and WT/L268P NBs, respectively. More importantly, the correct LLPS-driven NB formation was the prerequisite for partner recruitment, post-translational modifications (PTMs), and PML-driven cellular regulations, such as ROS stress control, mitochondria production, and PML-p53-mediated senescence and apoptosis. Altogether, our results helped to define a critical LLPS step in PML NB biogenesis.

A comprehensive benchmarking of differential splicing tools for RNA-seq analysis at the event level.

RNA alternative splicing, a post-transcriptional stage in eukaryotes, is crucial in cellular homeostasis and disease processes. Due to the rapid development of the next-generation sequencing (NGS) technology and the flood of NGS data, the detection of differential splicing from RNA-seq data has become mainstream. A range of bioinformatic tools has been developed. However, until now, an independent and comprehensive comparison of available algorithms/tools at the event level is still lacking. Here, 21 different tools are subjected to systematic evaluation, based on simulated RNA-seq data where exact differential splicing events are introduced. We observe immense discrepancies among these tools. SUPPA, DARTS, rMATS and LeafCutter outperforme other event-based tools. We also examine the abilities of the tools to identify novel splicing events, which shows that most event-based tools are unsuitable for discovering novel splice sites. To improve the overall performance, we present two methodological approaches i.e. low-expression transcript filtering and tool-pair combination. Finally, a new protocol of selecting tools to perform differential splicing analysis for different analytical tasks (e.g. precision and recall rate) is proposed. Under this protocol, we analyze the distinct splicing landscape in the DUX4/IGH subgroup of B-cell acute lymphoblastic leukemia and uncover the differential splicing of TCF12. All codes needed to reproduce the results are available at https://github.com/mhjiang97/Benchmarking_DS.

Metamifop resistance in Echinochloa glabrescens via glutathione S-transferases-involved enhanced metabolism.

BACKGROUND: Echinochloa glabrescens Munro ex Hook. f. is one of the main Echinochloa spp. seriously invading Chinese rice fields and has evolved resistance to commonly used herbicides. Previously, an E. glabrescens population (LJ-02) with suspected resistance to the acetyl-CoA carboxylase (ACCase)-inhibiting herbicide metamifop was collected. This study aimed to determine its resistance status to metamifop and investigate the internal molecular mechanisms of resistance. RESULTS: Single-dose testing confirmed that the LJ-02 population had evolved resistance to metamifop. Gene sequencing and a relative expression assay of ACCase ruled out target-site based resistance to metamifop in LJ-02. Whole-plant bioassays revealed that, compared with the susceptible population XZ-01, LJ-02 was highly resistant to metamifop and exhibited cross-resistance to fenoxaprop-P-ethyl. Pretreatment with the known glutathione S-transferase (GST) inhibitor, 4-chloro-7-nitrobenzoxadiazole (NBD-Cl), largely reversed the resistance to metamifop by approximately 81%. Liquid chromatography-tandem mass spectrometry analysis indicated that the metabolic rates of one of the major metabolites of metamifop, N-(2-fluorophenyl)-2-hydroxy-N-methylpropionamide (HPFMA), were up to 383-fold faster in LJ-02 plants than in XZ-01 plants. There were higher basal and metamifop-inducible GST activities toward 1-chloro-2,4-dinitrobenzene (CDNB) in LJ-02 than in XZ-01. Six GST genes were metamifop-induced and overexpressed in the resistant LJ-02 population. CONCLUSION: This study reports, for the first time, the occurrence of metabolic metamifop resistance in E. glabrescens worldwide. The high-level metamifop resistance in the LJ-02 population may mainly involve specific isoforms of GSTs that endow high catalytic activity and strong substrate specificity. © 2023 Society of Chemical Industry.

Primary assessment of the diversity of Omicron sublineages and the epidemiologic features of autumn/winter 2022 COVID-19 wave in Chinese mainland.

With the recent ongoing autumn/winter 2022 COVID-19 wave and the adjustment of public health control measures, there have been widespread SARS-CoV-2 infections in Chinese mainland. Here we have analyzed 369 viral genomes from recently diagnosed COVID-19 patients in Shanghai, identifying a large number of sublineages of the SARS-CoV-2 Omicron family. Phylogenetic analysis, coupled with contact history tracing, revealed simultaneous community transmission of two Omicron sublineages dominating the infections in some areas of China (BA.5.2 mainly in Guangzhou and Shanghai, and BF.7 mainly in Beijing) and two highly infectious sublineages recently imported from abroad (XBB and BQ.1). Publicly available data from August 31 to November 29, 2022 indicated an overall severe/critical case rate of 0.035% nationwide, while analysis of 5706 symptomatic patients treated at the Shanghai Public Health Center between September 1 and December 26, 2022 showed that 20 cases (0.35%) without comorbidities progressed into severe/critical conditions and 153 cases (2.68%) with COVID-19-exacerbated comorbidities progressed into severe/critical conditions. These observations shall alert healthcare providers to place more resources for the treatment of severe/critical cases. Furthermore, mathematical modeling predicts this autumn/winter wave might pass through major cities in China by the end of the year, whereas some middle and western provinces and rural areas would be hit by the upcoming infection wave in mid-to-late January 2023, and the duration and magnitude of upcoming outbreak could be dramatically enhanced by the extensive travels during the Spring Festival (January 21, 2023). Altogether, these preliminary data highlight the needs to allocate resources to early diagnosis and effective treatment of severe cases and the protection of vulnerable population, especially in the rural areas, to ensure the country's smooth exit from the ongoing pandemic and accelerate socio-economic recovery.

Effects of glycation methods on the interfacial behavior and emulsifying performance of soy protein isolate-gum Arabic conjugates.

Glycated conjugation of plant protein such as soy protein isolate (SPI) with saccharides is one popular strategy to modify the physicochemical characteristics of these plant protein resources, which may be affected by the glycation methods including dry-heating and wet-heating. In this study, the impact of these two glycation methods on the rheological and emulsifying properties of a binary system made by SPI-gum Arabic (GA) was studied. The results indicated that dry-heating conjugates had higher viscosity and more elastic characteristics than those wet-heating conjugates. The emulsifying properties of SPI-GA conjugates by different preparation routes were evaluated by various oil phases including eugenol, cinnamaldehyde and soybean oil. Overall, emulsions stabilized by dry-heating conjugates showed lower zeta-potential value than those with wet heating conjugates. The interfacial properties of these conjugates were compared using soybean oil emulsion as a model. Higher emulsifying ability and stability were obtained by emulsions with dry-heating conjugates, which was attributed to their more compact structures, higher protein adsorption capacity and thicker viscoelastic films formed at the interface, and therefore enhanced electrostatic repulsion between droplets. The findings in this study are useful for fabrication and utilization of protein-polysaccharide glycation conjugates as emulsifiers in functional foods.

Mathematical appraisal of SARS-CoV-2 Omicron epidemic outbreak in unprecedented Shanghai lockdown.

The SARS-CoV-2 Omicron outbreak is ongoing in Shanghai, home to 25 million population. Here, we presented a novel mathematical model to evaluate the Omicron spread and Zero-COVID strategy. Our model provided important parameters, the average quarantine ratio, the detection interval from being infected to being tested positive, and the spreading coefficient to understand the epidemic progression better. Moreover, we found that the key to a relatively accurate long-term forecast was to take the variation/relaxation of the parameters into consideration based on the flexible execution of the quarantine policy. This allowed us to propose the criteria for estimating the parameters and outcome for the ending stage that is likely to take place in late May. Altogether, this model helped to give a correct mathematical appraisal of the SARS-CoV-2 Omicron outbreak under the strict Zero-COVID policy in China.

Effect of Cationic Modified Microcrystalline Cellulose on the Emulsifying Properties and Water/Oil Interface Behavior of Soybean Protein Isolate.

Stabilizing emulsion using complex biopolymers is a common strategy. It would be very interesting to characterize the impact of charge density on the emulsifying properties of complex polyelectrolytes carrying opposite charges. In this study, cationic modified microcrystalline celluloses (CMCC) of different charge densities were prepared and mixed with soy protein isolate (SPI) for emulsion applications. CMCC-1 to 3 with various cationic charge values were successfully prepared as characterized by zeta-potential and FTIR. The positive charge density's effects on solubility, thermogravimetric properties, and rheological properties were studied. Complexes of SPI-CMCC with various zeta-potential values were then obtained and used to stabilize soybean oil emulsions. The results show that emulsions stabilized by complexes of SPI and CMCC-3 at a ratio of 1:3 had the best emulsification ability and stability. However, the interfacial tension-reducing ability of complexes decreased continuously with increasing cationic charge value, while the rheological results show that complexes of SPI-CMCC-3 at a ratio of 1:3 formed a stronger viscoelastic network than other complexes. Our results indicate that this SPI-CMCC complex formula showed excellent emulsification performance, which could be adjusted and promoted by changing the charge density. This complex formula is promising for fabrication of emulsion-based food and cosmetic products.

Targeted isolation of diterpenoids and sesquiterpenoids from Daphne gemmata E. Pritz. ex Diels using molecular networking together with network annotation propagation and MS2LDA.

Investigation of the whole plant of Daphne gemmata E. Pritz. ex Diels (Thymelaeaceae) using molecular networking coupled to Network Annotation Propagation (NAP) and unsupervised substructure annotation (MS2LDA) led to the discovery of five tigliane diterpenoids, 14 guaiane sesquiterpenoids, one rhamnofolane diterpenoid and three carotene sesquiterpenoids. The structures of the eight undescribed compounds, daphnorbol A and daphnegemmatoids A-G, were characterized by detailed spectroscopic analyses, NMR and ECD calculations, application of Snatzke's method and single-crystal X-ray diffraction analysis. All isolated compounds were evaluated for their cytotoxic activities against HepG2, A549, and MCF-7 cells by MTT assay. Daphnorbol A exhibited significant cytotoxic activity against HepG2 and A549 cells with IC50 values of 4.06 µM and 6.35 µM, respectively. Prostratin showed potent cytotoxic activity against HepG2 and A549 cells with IC50 values of 6.06 µM and 5.45 µM, respectively. Further Hoechst 33,258 and AO-EB staining assays indicated that daphnorbol A and prostratin could induce apoptosis in HepG2 and A549 cells.

Quality Changes in Black Chokeberry Juice Treated by Thermal-Assisted High Hydrostatic Pressure during Cold Storage.

The effects of thermal-assisted high hydrostatic pressure (TAHHP), high hydrostatic pressure (HHP), and thermal pasteurization (TP) treatments on the quality of aronia juice were evaluated in this study. The results showed that TAHHP and HHP significantly decreased the aerobic plate counts of aronia juice. No significant differences in terms of physicochemical properties, such as pH and total soluble solids, were observed between aronia juice treated with high pressure or thermal pasteurization treatment after 28 days of storage. TAHHP and HHP affected the colour and antioxidant characteristics of aronia juice, though to a significantly lower extent than TP. This result demonstrates that TAHHP and HHP can better maintain the original quality of aronia juice than TP. In summary, both TAHHP and HHP can maintain the microbiological safety and original quality characteristics of aronia juice. TAHHP can effectively increase the safety and duration of cold storage of aronia juice, and hence is highly useful for the juice industry.

Investigating resistance levels to cyhalofop-butyl and mechanisms involved in Chinese sprangletop (Leptochloa chinensis L.) from Anhui Province, China.

Chinese sprangletop (Leptochloa chinensis (L.) Nees) is a common grass species that severely threatens rice (Oryza sativa L.) cropping systems globally. Cyhalofop-butyl is a highly efficient acetyl-CoA carboxylase (ACCase)-inhibiting herbicide widely used for control of this species in China. However, some L. chinensis populations have gradually evolved resistance to this herbicide in recent years. To better understand the cyhalofop-butyl resistance status of L. chinensis in the major rice planting area of the middle-lower Yangtze River basin, 73 populations collected from the rice fields across Anhui Province were investigated for cyhalofop-butyl susceptibility and potential herbicide resistance-conferring mutations. Single-dose testing indicated that of the 73 populations, 25 had evolved resistance to cyhalofop-butyl and were separately classified as "RRR" and "RR" populations according to their fresh weight reductions, 8 had a high risk of evolving cyhalofop-butyl resistance and were classified as "R?" populations, and 40 were susceptible and classified as "S" populations. Whole-plant dose-response experiments showed that the resistance index (RI) of these R?, RR, and RRR populations to cyhalofop-butyl ranged from 2.47 to 36.94. Target gene sequencing identified seven ACCase resistance mutations (I1781L, W1999C, W2027S, W2027L, W2027C, I2041N, and D2078G), with W1999C and W2027C the two most common detected in about three quarters of all the resistant populations. Seven populations including LASC3, BBHY1, AQQS1, HFFD3, HFFD4, AQWJ1, and HFLJ6 each carrying a specific ACCase mutation were tested for their cross- and multiple-resistance patterns. Compared with a standard susceptible population HFLY1, the seven resistant populations showed distinct cross-resistance. All had low- to high-level cross-resistance to metamifop (RIs ranging from 6.16 to 17.65), fenoxaprop-P-ethyl (RIs ranging from 6.39 to 24.08), and quizalofop-P-ethyl (RIs ranging from 2.20 to 10.25), but responded differently to clodinafop-propargyl and clethodim. Multiple-resistance testing suggested that the seven resistant populations were all susceptible to the 4-hydroxyphenylpyruvate dioxygenase inhibitor tripyrasulfone, the protoporphyrinogen oxidase inhibitor oxyfluorfen, and the auxin mimic herbicide florpyrauxifen. In conclusion, this study has shown that cyhalofop-butyl resistance was prevalent in L. chinensis in Anhui Province, China, and target site mutation was one of the most common resistance mechanisms.

Functional, structural, and molecular characterizations of the leukemogenic driver MEF2D-HNRNPUL1 fusion.

Recurrent MEF2D fusions with poor prognosis have been identified in B-cell precursor ALL (BCP-ALL). The molecular mechanisms underlying the pathogenic function of MEF2D fusions are poorly understood. Here, we show that MEF2D-HNRNPUL1 (MH) knock-in mice developed a progressive disease from impaired B-cell development at the pre-pro-B stage to pre-leukemia over 10 to 12 months. When cooperating with NRASG12D, MH drove an outbreak of BCP-ALL, with a more aggressive phenotype than the NRASG12D-induced leukemia. RNA-sequencing identified key networks involved in disease mechanisms. In chromatin immunoprecipitation-sequencing experiments, MH acquired increased chromatin-binding ability, mostly through MEF2D-responsive element (MRE) motifs in target genes, compared with wild-type MEF2D. Using X-ray crystallography, the MEF2D-MRE complex was characterized in atomic resolution, whereas disrupting the MH-DNA interaction alleviated the aberrant target gene expression and the B-cell differentiation arrest. The C-terminal moiety (HNRNPUL1 part) of MH was proven to contribute to the fusion protein's trans-regulatory activity, cofactor recruitment, and homodimerization. Furthermore, targeting MH-driven transactivation of the HDAC family by using the histone deacetylase inhibitor panobinostat in combination with chemotherapy improved the overall survival of MH/NRASG12D BCP-ALL mice. Altogether, these results not only highlight MH as an important driver in leukemogenesis but also provoke targeted intervention against BCP-ALL with MEF2D fusions.

Expression Pattern of Entire Cytochrome P450 Genes and Response of Defenses in a Metabolic-Herbicide-Resistant Biotype of Polypogon fugax.

Enhanced herbicide metabolism mediated by cytochrome P450s has been proposed as one of the major mechanisms of resistance to fenoxaprop-P-ethyl in a metabolic-herbicide-resistant biotype of Asia minor bluegrass (Polypogon fugax Nees ex Steud.). Upon pre-treatment with the P450 inhibitor piperonyl butoxide, a remarkable reduction in metabolic rates of the phytotoxic fenoxaprop-P has been observed in the resistant plants, implying that constitutive and/or fenoxaprop-P-ethyl-induced up-regulation of specific P450 isoforms are involved in the fenoxaprop-P-ethyl resistance. However, which P450 gene(s) were responsible for the metabolic resistance is still unknown. In this present study, based on the abundant gene resources of P. fugax established previously, a total of 48 putative P450 genes were isolated from the metabolic-herbicide-resistant plants and used for gene expression analysis. The most suitable reference genes for accurate normalization of real-time quantitative PCR data were first identified in P. fugax and recognized as actin (ACT), 18S rRNA (18S), and ribulose-1,5-bisphosphate carboxylase oxygenase (RUBP) under fenoxaprop-P-ethyl stress, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and elongation factor 1α (EF1α) under mesosulfuron-methyl stress, and ACT, EF1α, eukaryotic initiation factor 4a (EIF4A), and 25S rRNA (25S) at different growth stages. Expression analysis of the putative P450 genes revealed that six genes, respectively, annotated as CYP709B1, CYP71A1-4, CYP711A1, CYP78A9, P450-11, and P450-39 were up-regulated more than 10-fold in the resistant plants by fenoxaprop-P-ethyl treatment, and all of them exhibited constitutively and/or herbicide-induced higher transcript levels in the fenoxaprop-P-ethyl-resistant than in the susceptible plants. Three genes, respectively, annotated as CYPRO4, CYP313A4, and CYP51H11 constantly up-regulated in the resistant than in the susceptible plants after fenoxaprop-P-ethyl treatment. Up-regulated expressions of these specific P450 genes were consistent with the higher P450 contents determined in the resistant plants. These results will help to elucidate the mechanisms for P450-mediated metabolic-herbicide resistance in P. fugax as well as other grass weed species.

Identification of essential genes involved in metabolism-based resistance mechanism to fenoxaprop-P-ethyl in Polypogon fugax.

BACKGROUND: Metabolic resistance is a worldwide concern for weed control but has not yet been well-characterized at the genetic level. Previously, we have identified an Asia minor bluegrass (Polypogon fugax Nees ex Steud.) population AHHY exhibiting cytochrome P450 (P450)-involved metabolic resistance to fenoxaprop-P-ethyl. In this study, we aimed to confirm the metabolic fenoxaprop-P-ethyl resistance in AHHY and uncover the potential herbicide metabolism-related genes in this economically damaging weed species. RESULTS: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays indicated the metabolic rates of fenoxaprop-P-ethyl were significantly faster in resistant (R, AHHY) than in susceptible (S, SDTS) plants. The amount of phytotoxic fenoxaprop-P peaked at 12 h after herbicide treatment (HAT) and started to decrease at 24 HAT in both biotypes. R and S plants at 24 HAT were sampled to conduct isoform-sequencing (Iso-Seq) and RNA-sequencing (RNA-Seq). A reference transcriptome containing 24 972 full-length isoforms was obtained, of which 24 329 unigenes were successfully annotated. Transcriptomic profiling identified 28 detoxifying enzyme genes constitutively and/or herbicide-induced up-regulated in R than in S plants. Real-time quantitative polymerase chain reaction (RT-qPCR) confirmed 17 genes were consistently up-regulated in R and its F1 generation plants. They were selected as potential fenoxaprop-P-ethyl metabolism-related genes, including ten P450s, one glutathione-S-transferase, one UDP-glucosyltransferase, and five adenosine triphosphate (ATP)-binding cassette transporters. CONCLUSION: This study revealed that the enhanced rates of fenoxaprop-P-ethyl metabolism in P. fugax were very likely driven by the herbicide metabolism-related genes. The transcriptome data generated by Iso-Seq combined with RNA-Seq will provide abundant gene resources for understanding the molecular mechanisms of resistance in P. fugax.