Is autophagy always a death sentence? A case study of highly selective cytoplasmic degradation during phloemogenesis.

BACKGROUND AND AIMS: The transformation of sieve elements (SEs) from meristematic cells, equipped with a full complement of organelles, to specialized transport tubes devoid of a nucleus, has long been enigmatic. We hypothesized a strong involvement of various degradation pathways, particularly macroautophagy in this context, emphasizing the importance of autophagic selectivity in the remaining viability of these cells. METHODS: Experiments were performed on pioneer roots of Populus trichocarpa cultivated in rhizotrons under field conditions. Through anatomical, ultrastructural and molecular analyses, we delineate the stages of phloemogenesis and the concurrent alterations in the cytoplasmic composition of SEs. KEY RESULTS: Notably, we observed not only macroautophagic structures, but also the formation of autophagic plastids, the selective degradation of specific organelles, vacuole disruption and the release of vacuolar contents. These events initially lead to localized reductions in cytoplasm density, but organelle-rich cytoplasmic phase is safeguarded from the extensive damage by a membrane system derived from the endoplasmic reticulum. SE ultimately develops into a conduit containing electron-translucent cytoplasm. Eventually, mature SE is a tube filled only by the translucent cytoplasm, with sparse organelles tethered to the cell wall. CONCLUSIONS: Although the activation of programmed cell death pathways was postulated, the persistence of SEs indicates that protoplast depletion is meticulously regulated by hitherto unidentified mechanisms. This research elucidates the sequential processes occurring in these cells during phloemogenesis and unveils novel insights into the mechanisms of selective autophagy.

Giant cystic Intradural extramedullary tumor of the lumbar spine mimicking arachnoid cyst: A case report.

INTRODUCTION: Schwannoma, also known as neurilemmoma, is a benign tumor commonly found around the spinal nerve roots. Large, solitary cases of cystic degeneration within the extramedullary intradural compartment (IDEM) can be challenging for preoperative diagnosis. Furthermore, these cases are not extensively documented in the medical literature. CASE PRESENTATION: We report a case of giant invasive IDEM schwannoma in a 28-year-old man who presented with intermittent pain radiating to the left thigh without numbness. Magnetic resonance imaging (MRI) showed a complex cystic lesion within the spinal canal from the inferior end plate of L4 to the mid-body of S1. CLINICAL DISCUSSION: The patient underwent a series of surgical procedures, including laminectomy and decompression at the L4-L5 level for resection of the tumor. Histopathological examination confirmed the diagnosis of schwannoma. The patient had a favorable postoperative recovery and experienced a resolution of symptoms. CONCLUSION: The giant cystic lumbar schwannoma, although rare, necessitates careful diagnosis and management. The utilization of contrast-enhanced MRI aids in distinguishing it from other lesions. In cases of cystic spinal schwannoma, the lesion walls tend to be thicker and more irregular compared to other cysts, including arachnoid cysts. Histopathological examination should be utilized to identify these lesions intraoperatively. Surgical excision is the primary treatment, and complete excision should be attempted whenever feasible.

Quantitative Proteomic Analysis of Brassica Napus Reveals Intersections Between Nutrient Deficiency Responses.

Macronutrients such as nitrogen (N), phosphorus (P), potassium (K) and sulphur (S) are critical for plant growth and development. Field-grown canola (Brassica napus L.) is supplemented with fertilizers to maximize plant productivity, while deficiency in these nutrients can cause significant yield loss. A holistic understanding of the interplay between these nutrient deficiency responses in a single study and canola cultivar is thus far lacking, hindering efforts to increase the nutrient use efficiency of this important oil seed crop. To address this, we performed a comparative quantitative proteomic analysis of both shoot and root tissue harvested from soil-grown canola plants experiencing either nitrogen, phosphorus, potassium or sulphur deficiency. Our data provide critically needed insights into the shared and distinct molecular responses to macronutrient deficiencies in canola. Importantly, we find more conserved responses to the four different nutrient deficiencies in canola roots, with more distinct proteome changes in aboveground tissue. Our results establish a foundation for a more comprehensive understanding of the shared and distinct nutrient deficiency response mechanisms of canola plants and pave the way for future breeding efforts.

Response of growth and chlorophyll fluorescence parameters of mulberry seedlings to waterlogging stress.

This study aimed to investigate the adaptive mechanisms of mulberry (Morus alba) to waterlogged conditions, with a specific focus on the development of adventitious roots (ARs), alteration of growth strategies, and adjustment of chlorophyll fluorescence parameters. To achieve this goal, 4-year-old potted mulberry plants were selected for research, and a waterlogging simulation method was implemented. Four treatments were established to investigate the effects of varying water conditions on leaf waterlogging damage, the number of ARs, plant height, chlorophyll fluorescence parameters, and proton motive force (pmf) parameters in mulberry plants. These treatments included the control group (CK), shallow submerged group (SS), half-submerged group (HS) and deep submerged group (DS). Our results showed that (1) The number of ARs in each group increased with increasing waterlogging time. (2) Waterlogging stress inhibited the height growth of mulberry, and the changes in plant height in the HS and DS groups were significantly lower than those in the CK and SS groups. (3) The maximum photochemical quantum yield (Fv/Fm) in the HS and DS groups decreased significantly under waterlogging stress. The nonphotochemical quenching (NPQt) of mulberry leaves in the submergence group increased significantly in the early stage of waterlogging stress, and the NPQt in the submergence group increased continuously with increasing waterlogging time. (4) Thylakoid conductivity to protons (gH+) in the leaves of mulberry decreased significantly under waterlogging stress, whereas the steady-state rate of proton flux (vH+) and total electrochromic shift (ECSt) increased significantly. The morphological, physiological, and ecological responses of mulberry plants to waterlogging stress include the timely generation of ARs at the stem base, the adjustment of plant growth strategies, and the repair of photosynthetic response centers in leaves through heat dissipation and thylakoid acidification mechanisms.

Variability in growth and biosynthetic activity of Calendula officinalis hairy roots.

Calendula officinalis is a widespread medicinal plant with a sufficiently well-studied chemical composition. Secondary metabolites synthesized by C.officinalis plants have pharmacological value for treating numerous diseases, and various types of aseptic in vitro cultures can be used as a source of these compounds. From this perspective, hairy roots attract considerable attention for the production of bioactive chemicals, including flavonoids with antioxidant activity. This paper shows the possibility of C.officinalis hairy roots obtaining with 100% frequency by Agrobacterium rhizogenes genetic transformation. Hairy root lines differed in growth rate and flavonoid content. In particular, flavonoids were accumulated in the amount of up to 6.68 ± 0.28 mg/g of wet weight. Methyl jasmonate in the concentration of 10 µM inhibited root growth to a small extent but stimulated the synthesis of flavonoids. The antioxidant activity and the reducing power increased in the roots grown in the medium with methyl jasmonate. The strong correlation of antioxidant activity and reducing power with flavonoid content was detected. The influence of extraction conditions on the content of flavonoids in the extracts and their bioactivity was determined. The potent reducing activity of extracts from hairy roots allowed the production of silver nanoparticles, which was confirmed by transmission electron microscopy.

Integrating UPLC-Q-Exactive Orbitrap/MS, Network pharmacology and experimental validation to reveal the potential mechanism of Kadsuracoccinea roots in Colon Cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: Kadsura coccinea roots are a traditional folk medicine used to treat gastrointestinal diseases. In recent years, research on K. coccinea has predominantly focused on the analysis of chemical composition and screening for activity, but there is a scarcity of studies that employ mass spectrometry to analyze Kadsura coccinea roots. AIM OF THE STUDY: This study aimed to characterize the chemical composition of K. coccinea roots and explore the pharmacological mechanisms with network pharmacology. Cell assay and Western blot analysis were used to verify the pharmacological mechanism of the main compounds in K. coccinea roots. MATERIALS AND METHODS: UPLC-Q-Exactive Orbitrap/MS was used for chemical analysis of K. coccinea roots, and the compounds were identified by employing diagnostic product ions, fragmentation patterns, ChemSpider, and in-house databases. Network pharmacology was employed to estimate the pathways related to pharmacological mechanisms. In addition, MTT assay was conducted to determine the inhibitory activity of colon cancer cell lines, and their apoptotic abilities were evaluated by flow cytometry and Western blot. RESULTS: The UPLC-Q-Exactive Orbitrap/MS identified a total of 54 compounds in K. coccinea roots. The 54 compounds were subjected to network pharmacology analysis, exploring the pharmacological action of the main components of K. coccinea roots. The common targets between the compounds and colon cancer comprised 2009 GO biological process items and 186 KEGG signal pathways. Flow cytometry indicated that treatments with 20 µM of the above-named compounds resulted in an apoptosis rate of 16.6%, 79.7%, and 22.2% in HCT-116 cells, respectively. Meanwhile, Western blot analysis confirmed that the compounds promoted the expression of Bax and Caspase-3 level expression. CONCLUSION: The findings demonstrated that K. coccinea roots can treat colon cancer through multiple components, targets, and pathways. This study revealed the effective components and molecular mechanisms of K. coccinea, which were preliminarily verified using in vitro experiments.

Response of Root Respiration to Warming and Nitrogen Addition Depends on Tree Species.

Roots contribute a large fraction of CO2 efflux from soils, yet the extent to which global change factors affect root-derived fluxes is poorly understood. We investigated how red maple (Acer rubrum) and red oak (Quercus rubra) root biomass and respiration respond to long-term (15 years) soil warming, nitrogen addition, or their combination in a temperate forest. We found that ecosystem root respiration was decreased by 40% under both single-factor treatments (nitrogen addition or warming) but not under their combination (heated × nitrogen). This response was driven by the reduction of mass-specific root respiration under warming and a reduction in maple root biomass in both single-factor treatments. Mass-specific root respiration rates for both species acclimated to soil warming, resulting in a 43% reduction, but were not affected by N addition or the combined heated × N treatment. Notably, the addition of nitrogen to warmed soils alleviated thermal acclimation and returned mass-specific respiration rates to control levels. Oak roots contributed disproportionately to ecosystem root respiration despite the decrease in respiration rates as their biomass was maintained or enhanced under warming and nitrogen addition. In contrast, maple root respiration rates were consistently higher than oak, and this difference became critical in the heated × nitrogen treatment, where maple root biomass increased, contributing significantly more CO2 relative to single-factor treatments. Our findings highlight the importance of accounting for the root component of respiration when assessing soil carbon loss in response to global change and demonstrate that combining warming and N addition produces effects that cannot be predicted by studying these factors in isolation.

Novel Colorimetric and Near-Infrared Ratiometric Fluorescent Probe for Sensing Cysteine in Food Samples, Plants, and Living Cells.

Cysteine (Cys) is a crucial biothiol that acts a significant function in food samples and biological systems, including plant roots and living cells. Hence, we developed a novel colorimetric and near-infrared ratiometric fluorescent probe (CT), composed of coumarin and tetrahydroacridine-conjugated indole salt, for the detection of Cys. Upon reaction with Cys, the probe undergoes a specific N-substitution reaction, resulting in a notable colorimetric change and a significant ratiometric fluorescent response in both visible and near-infrared emission channels. These dual-channel ratiometric fluorescence changes are completely independent, enabling the probe to obtain great selectivity, sensitivity, and exceptional detection accuracy. Leveraging these attributes, the probe was employed to provide accurate quantitative analysis of Cys in food samples. Furthermore, confocal imaging demonstrated that the probe could monitor both exogenous and endogenous Cys levels in living cells and track Cys changes in plant roots under heavy metal stress. This work presents a dependable and accurate imaging solution for tracking and identifying Cys of real food, plants, and living cells.

The DTR Technique-Drilling through the Roots of Posterior Teeth for Anatomically Guided Immediate Implant Placement: A Cohort Study.

AIM: The present study clinically analyzes implant survival of immediate implant placement cases using the drilling through roots (DTR) technique for anatomically-guided implant site preparation, as an aid to placing immediate dental implants in multi-radicular teeth. MATERIALS AND METHODS: This clinical analysis utilized patients' electronic dental records who underwent immediate implant surgery using the DTR technique. All immediately placed implants were followed up regularly every year, after restoration. Implant survival was assessed with the Albrektsson et al. criteria. Inferential statistics was performed using SPSS v 21(IBM Corp., Armonk, NY) software. The Kaplan-Meier survival analysis was done to assess the implant survival probability. RESULTS: A total of 250 records of dental implants placed in 227 subjects using the DTR technique were considered. Results showed that the mean survival duration of implants was found 63.29 months and the median survival duration to be 55 months. A 100% success rate was seen in implant fixed bridge cases, and about 97.6% success was seen in single crown cases. No significant difference was seen in the survival rates during the follow-up period when compared according to the quadrants/site of implant placement. CONCLUSION: The findings concluded that tooth-guided rapid implant placement is a unique strategy for convenient and safe insertion, providing accurate three-dimensional positioning. CLINICAL SIGNIFICANCE: The DTR method is a novel approach that facilitates accurate positioning and angulation of the implant bed preparation by stabilizing and guiding the osteotomy drills using the retained root. As a result, it enables optimal implant positioning at multirooted extraction sites. How to cite this article: Mahesh L, Miselli A, Bhasin MT, et al. The DTR Technique-Drilling through the Roots of Posterior Teeth for Anatomically Guided Immediate Implant Placement: A Cohort Study. J Contemp Dent Pract 2024;25(5):432-439.

Divide and conquer: using RhizoVision Explorer to aggregate data from multiple root scans using image concatenation and statistical methods.

Roots are important in agricultural and natural systems for determining plant productivity and soil carbon inputs. Sometimes, the amount of roots in a sample is too much to fit into a single scanned image, so the sample is divided among several scans, and there is no standard method to aggregate the data. Here, we describe and validate two methods for standardizing measurements across multiple scans: image concatenation and statistical aggregation. We developed a Python script that identifies which images belong to the same sample and returns a single, larger concatenated image. These concatenated images and the original images were processed with RhizoVision Explorer, a free and open-source software. An R script was developed, which identifies rows of data belonging to the same sample and applies correct statistical methods to return a single data row for each sample. These two methods were compared using example images from switchgrass, poplar, and various tree and ericaceous shrub species from a northern peatland and the Arctic. Most root measurements were nearly identical between the two methods except median diameter, which cannot be accurately computed by statistical aggregation. We believe the availability of these methods will be useful to the root biology community.

Weighted gene co-expression network analysis and identification of ginsenoside biosynthesis candidate genes for ginseng adventitious roots under MeJA treatment.

BACKGROUND: Ginseng (Panax ginseng) is an herb with a long history and a wide range of applications. Ginsenoside is one of the most representative and active ginseng compounds, with various pharmacological effects. Therefore, the development of bioreactors using methyl jasmonate (MeJA) as an inducer for targeted ginsenoside production is of great commercial value. Combined with transcriptomic research tools, screenings to obtain candidate genes involved in ginsenoside biosynthesis are crucial for future discoveries about the molecular mechanism of MeJA-regulated ginsenoside biosynthesis. OBJECTIVE AND METHODS: In our study, the ginsenoside content of ginseng adventitious roots treated with MeJA at different times was analyzed. Transcriptome analysis was performed to investigate the effects of MeJA on changes in ginsenoside content in ginseng adventitious roots. RESULTS: The MeJA could significantly increase changes in the content of pro-ginsenodiol ginsenosides as well as pro-triol ginsenosides Rg3, Re, and Rf in ginseng adventitious roots. Differential gene expression analysis showed that a total of 14,009 differentially expressed genes were obtained from the screening of the present study. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that differentially expressed genes were mainly enriched under GO terms in response to stimuli, metabolic processes, and the regulation of biological processes, with significant annotation to the metabolic terms of terpenoids and polyketides. Two expression modules of genes highly related to ginsenoside biosynthesis were obtained via WGCNA. CONCLUSIONS: Our study provides a reference system for the targeted ginsenoside production using MeJA as an inducer, and also provides genetic and gene resources for subsequently validating genes related to the regulation of ginsenoside biosynthesis using weighted gene co-expression network analysis (WGCNA).

Access Pain During Transforaminal Endoscopic Lumbar Discectomy for Foraminal or Extraforaminal Disc Herniation.

BACKGROUND/OBJECTIVES: Transforaminal endoscopic lumbar discectomy (TELD) under local anesthesia is a promising minimally invasive surgical option for intractable lumbar disc herniation (LDH). However, our understanding of access pain prediction during foraminal pathological procedures is limited. To our knowledge, no predictive rules for access pain have been established during TELD for foraminal or extraforaminal LDH. This study, with its potential for predicting access pain during TELD and discussing strategies for pain prevention and management, could significantly benefit the field of endoscopic spine surgery. METHODS: This observational study included 73 consecutive patients who underwent TELD for foraminal or extraforaminal LDH between January 2017 and December 2022. Preoperative clinical and radiographic factors affecting significant access pain and the impact of access pain on clinical outcomes were evaluated. RESULTS: The rate of significant access pain was 13.70% (10 of 73 patients). Extraforaminal LDH tended to cause more severe pain than did foraminal LDH during TELD under local anesthesia (p < 0.05). Although the degree of access pain was not related to global clinical outcomes, increased pain was strongly associated with prolonged operative time and length of hospital stay (p < 0.05). CONCLUSIONS: TELD could be an effective surgical option for foraminal or extraforaminal LDH under local anesthesia. More access pain might develop during TELD for extraforaminal LDH. The extraforaminal component of LDH could narrow the safe working zone. Significant access pain might prolong the duration of surgery and hospitalization. Thus, a specialized technique is required for the clinical success of TELD.

GmSTOP1-3 regulates flavonoid synthesis to reduce ROS accumulation and enhance aluminum tolerance in soybean.

Aluminum (Al) toxicity is a significant limiting factor for crop production in acid soils. The functions and regulatory mechanisms of transcription factor STOP1 (Sensitive to Proton Rhizotoxicity 1) family genes in Al-tolerance have been widely studied in many plant species, except for soybean. Here, expression of GmSTOP1-3 was significantly enhanced by Al stress in soybean roots. Overexpression of GmSTOP1-3 resulted in enhanced root elongation and decreased Al content, which was accompanied by increased antioxidant capacity under Al treatment. Furthermore, RNA-seq identified 498 downstream genes of GmSTOP1-3, including genes involved in flavonoid biosynthesis. Among them, the expression of chalcone synthase (GmCHS) and isoflavone synthase (GmIFS) were highly enhanced by GmSTOP1-3 overexpression. Further quantitative flavonoid metabolome analysis showed that overexpression of GmSTOP1-3 significantly increased the content of naringenin chalcone, naringenin, and genistein in soybean roots under Al treatment, which positively correlated with the expression level of the genes relative to flavonoid biosynthesis. Notably, genistein had a significant positive correlation with the expression levels of GmIFS. Combination of Dual Luciferase Complementation (LUC) and Electrophoretic Mobility Shift Assays (EMSA) revealed that GmSTOP1-3 directly bound to the promoters of GmCHS/GmIFS and activated both genes' transcription. Taken together, these results suggest that GmSTOP1-3 enhances soybean Al tolerance partially through regulating the flavonoid synthesis.

Multi-scale phenotyping of senescence-related changes in roots of Rapeseed in response to nitrate limitation.

Root senescence remains largely unexplored. In this study, the temporality of the morphological, metabolic, and proteomic changes occurring with root aging were investigated, providing a comprehensive picture of the root senescence program. We found novel senescence-related markers for the characterization of the developmental stage of root tissues. The rapeseed root system is unique in that it consists of the taproot and lateral roots. Our study confirms that the taproot, which transiently accumulates large quantities of starch and proteins, is specifically dedicated to nutrient storage and remobilization, while the lateral roots are mainly dedicated to nutrient uptake. Proteomic data from the taproot and lateral roots highlight the different senescence-related events that control nutrient remobilization and nutrient uptake capacities. Both the proteome and enzyme activities revealed senescence-induced proteases and nucleotide catabolic enzymes that deserve attention as they may play important roles in nutrient remobilization efficiency in rapeseed roots. Taking advantage of publicly available transcriptomic and proteomic data on senescent Arabidopsis leaves, we have highlighted new lists of senescence-related proteins specific or common to root organs and/or leaves.

Pectin from comfrey roots alleviate DSS-induced ulcerative colitis in mice through modulating the intestinal barrier.

The objective of this study was to investigate whether pectin extracted from comfrey roots (CRPs) could alleviate ulcerative colitis (UC) induced by sodium dextran sulfate (DSS) in mice by improving the intestinal barrier. CRP was able to relieve symptoms associated with weight loss, diarrhea, and colon length in UC mice. CRP inhibited the excessive secretion of TNF-α and IL-6 and increased the level of IL-10 in the serum. After CRP treatment, the mRNA expression levels of ZO-1 and Muc2 increased. The colonic epithelial cells recovered well, and the mucous layer was relatively intact in the CRP group. CRP and 5-aminosalicylic acid (ASA) alleviated UC symptoms in mice by reducing the abundance of Oscillibacter, Alistipes, and Anaeroplasma and increasing the abundance of Lachnospiraceae_UCG-001 to regulate the intestinal microflora. The abundance of Rikenellaceae was positively correlated with the mRNA expression level of ZO-1, and the abundance of Monoglobus was positively correlated with the mRNA expression level of Muc2. These results suggested that CRP could repair the intestinal barrier and mitigate DSS-induced colon damage in mice, indicating that CRP may be a potential functional component in combating UC.

Comprehensive characterization of gibberellin oxidase gene family in Brassica napus reveals BnGA2ox15 involved in hormone signaling and response to drought stress.

Brassica napus is a well-known allopolyploid oil crop with high commercial potential. Gibberellin oxidase (GAox) is an essential enzyme that activates gibberellins, which regulate plant growth, and development, and have a significant impact on plant responses to abiotic stress. However, the comprehensive understanding of GAox genes and their evolution in Brassica plants remains elusive. Using advanced bioinformatics tools, this study identified 125 candidate GAox genes from the whole genomes of three key Brassica species. This study also investigated sequence characteristics, conserved motifs, exon/intron structures, cis-acting elements, syntenic analysis, duplication events and expression patterns. Subcellular localization analysis showed that the BnGA2ox14 and BnGA2ox15 proteins are located in the nucleus, whereas BnGA2ox26 is specifically localized to the chloroplast. Yeast one-hybrid and dual-luciferase assays demonstrated that MYELOCYTOMATOSIS 4 (BnMYC4) and ABA-INDUCIBLE BHLH-TYPE TRANSCRIPTION FACTOR (BnAIB) bind to the BnGA2ox15 promoter and activate its transcription. Molecular docking analysis further elucidated their interaction structures and identified potential binding sites. Roots transformations show that overexpression of BnGA2ox15 increased sensitivity to PEG-6000 treatment in rapeseed. In brief, this study reveals that BnGA2ox15 is a downstream target in JA and ABA signaling pathways, functioning as a negative regulator in response to drought stress.

Variation in forest root image annotation by experts, novices, and AI.

BACKGROUND: The manual study of root dynamics using images requires huge investments of time and resources and is prone to previously poorly quantified annotator bias. Artificial intelligence (AI) image-processing tools have been successful in overcoming limitations of manual annotation in homogeneous soils, but their efficiency and accuracy is yet to be widely tested on less homogenous, non-agricultural soil profiles, e.g., that of forests, from which data on root dynamics are key to understanding the carbon cycle. Here, we quantify variance in root length measured by human annotators with varying experience levels. We evaluate the application of a convolutional neural network (CNN) model, trained on a software accessible to researchers without a machine learning background, on a heterogeneous minirhizotron image dataset taken in a multispecies, mature, deciduous temperate forest. RESULTS: Less experienced annotators consistently identified more root length than experienced annotators. Root length annotation also varied between experienced annotators. The CNN root length results were neither precise nor accurate, taking ~ 10% of the time but significantly overestimating root length compared to expert manual annotation (p = 0.01). The CNN net root length change results were closer to manual (p = 0.08) but there remained substantial variation. CONCLUSIONS: Manual root length annotation is contingent on the individual annotator. The only accessible CNN model cannot yet produce root data of sufficient accuracy and precision for ecological applications when applied to a complex, heterogeneous forest image dataset. A continuing evaluation and development of accessible CNNs for natural ecosystems is required.

Accumulation of nanoplastics by wheat seedling roots: Both passive and energy-consuming processes.

Nanoplastics can transfer from the environment to plants and potentially harm organisms. However, the mechanisms on how crop root systems absorb and transport nanoplastics are still unclear. Here, original and fluorescent labeled polystyrene and polyvinyl chloride nanoparticles (PS-NPs, PVC-NPs; 30 nm; 10 mg L-1) were employed to study the distribution and internalization pathways in wheat seedling roots. In the study, nanoplastics accumulated more in the root tip and surface, with PVC-NPs more prevalent than PS-NPs. After being treated with inhibitors (Na3VO4, chlorpromazine and amiloride), the nanoplastics mean fluorescence intensities were reduced by 4.0-51.1 %. During the uptake, both passive and energy-consuming pathways occurred. For the energy-consuming uptake pathway, macropinocytosis contributed more to cytoplasm than clathrin-mediated endocytosis. H+ influx was observed during nanoplastic transport into the cytoplasm, and the reduction in plasma membrane ATPase activity led to a decrease in nanoplastic internalization. These results elucidate the pathways of nanoplastics absorption and transport in wheat roots, provide crucial evidence for assessing nanoplastics' ecological risks and support the development of technologies to block nanoplastics absorption by crop roots, ensuring agricultural and ecosystem safety.

Carbon dot unravels accumulation of triterpenoid in Evolvulus alsinoides hairy roots culture by stimulating growth, redox reactions and ANN machine learning model prediction of metabolic stress response.

Evolvulus alsinoides, a therapeutically valuable shrub can provide consistent supply of secondary metabolites (SM) with pharmaceutical significance. Nonetheless, because of its short life cycle, fresh plant material for research and medicinal diagnostics is severely scarce throughout the year. The effects of exogenous carbon quantum dot (CD) application on metabolic profiles, machine learning (ML) prediction of metabolic stress response, and SM yields in hairy root cultures of E. alsinoides were investigated and quantified. The range of the particle size distribution of the CDs was between 3 and 7 nm. The CDs EPR signal and spin trapping experiments demonstrated the formation of O2-•spin-adducts at (g = 2.0023). Carbon dot treatment increased the levels of hydrogen peroxide and malondialdehyde concentrations as well as increased antioxidant enzyme activity. CD treatments (6 µg mL-1) significantly enhanced the accumulation of squalene and stigmasterol (7 and 5-fold respectively). The multilayer perceptron (MLP) algorithm demonstrated remarkable prediction accuracy (MSE value = 1.99E-03 and R2 = 0.99939) in both the training and testing sets for modelling. Based on the prediction, the maximum oxidative stress index and enzymatic activities were highest in the medium supplemented with 10 µg mL-1 CDs. The outcome of this study indicated that, for the first time, using CD could serve as a novel elicitor for the production of valuable SM. MLP may also be used as a forward-thinking tool to optimize and predict SM with high pharmaceutical significance. This study would be a touchstone for understanding the use of ML and luminescent nanomaterials in the production and commercialization of important SM.

DNA Hypomethylation Activates the RpMYB2-Centred Gene Network to Enhance Regeneration of Adventitious Roots.

Plants, being immobile, are exposed to environmental adversities such as wind, snow and animals that damage their structure, making regeneration essential for their survival. The adventitious roots (ARs) primarily emerge from a detached explant to uptake nutrients; therefore, the molecular network involved in their regeneration needs to be explored. DNA methylation, a key epigenetic mark, influences molecular pathways, and recent studies suggested its role in regeneration. In our research, the application of 5-azacytidine (5-azaC), an inhibitor of DNA methylation, caused the earlier initiation and development of root primordia and consequently enhanced the AR regeneration rate in Robinia psuedoacacia L (black locust). The whole-genome bisulfite sequencing (WGBS) revealed a decrease in global methylation and an increase in hypomethylated cytosine sites and regions across all contexts including CHH, CHG and mergedCG caused transcriptional variations in 5-azaC-treated sample. The yeast two-hybrid (Y2H) assay revealed a RpMYB2-centred network of transcriptionally activated transcription factors (TFs) including RpWRKY23, RpGATA23, RpSPL16 and other genes like RpSDP, RpSS1, RpBEN1, RpGULL05 and RpCUV with nuclear localization suggesting their potential co-localization. Additionally, yeast one-hybrid (Y1H) assay showed the interaction of RpMYB2 interactors, RpGATA23 and RpWRKY23, with promoters of RpSK6 and RpCDC48, and luciferase reporting assay (LRA) validated their binding with RpSK6. Our results revealed that hypomethylation-mediated transcriptomic modifications activated the RpMYB2-centred gene network to enhance AR regeneration in black locust hypocotyl cuttings. These findings pave the way for genetic modification to improve plant regeneration ability and increase wood production while withstanding environmental damage.