Toxicity of triclosan, an antimicrobial agent, to a nontarget freshwater zooplankton species, Moina macrocopa.

The toxicity of triclosan (TCS) on the freshwater cladoceran Moina macrocopa was investigated by acute and chronic toxicity assessments followed by genotoxicity and oxidative stress response analyses. The 48-h LC50 of TCS for ≤24-h-old M. macrocopa was determined as 539 µg L-1 . Chronic exposure to TCS at concentrations ranging from 5 to 100 µg L-1 showed a stimulatory effect at low concentrations (≤10 µg L-1 ) and an inhibitory effect at high concentrations (≥50 µg L-1 ) on growth, reproduction, and population-growth-related parameters of M. macrocopa. The genotoxicity test results indicated that TCS concentrations ranging from 50 to 100 µg L-1 can alter individuals' DNA. Analysis of the antioxidant enzymes catalase (CAT) and glutathione s-transferase (GST) demonstrated increased levels of these enzymes at high TCS concentrations. Our results indicated that TCS concentrations found in the natural environment have minimal acute toxicity to M. macrocopa. However, TCS at even low concentrations can significantly affect its growth, reproduction, and population-growth-related characteristics. The observed responses suggest a hormetic dose-response pattern and imply a potential endocrine-disrupting effect of TCS. Our molecular and biochemical findings indicated that high concentrations of TCS have the potential to induce oxidative stress that may lead to DNA alterations in M. macrocopa.

Mechanisms involved in hematopoietic stem cell aging.

Hematopoietic stem cells (HSCs) undergo progressive functional decline over time due to both internal and external stressors, leading to aging of the hematopoietic system. A comprehensive understanding of the molecular mechanisms underlying HSC aging will be valuable in developing novel therapies for HSC rejuvenation and to prevent the onset of several age-associated diseases and hematological malignancies. This review considers the general causes of HSC aging that range from cell-intrinsic factors to cell-extrinsic factors. In particular, epigenetics and inflammation have been implicated in the linkage of HSC aging, clonality, and oncogenesis. The challenges in clarifying mechanisms of HSC aging have accelerated the development of therapeutic interventions to rejuvenate HSCs, the major goal of aging research; these details are also discussed in this review.

Backdoor Attack on Deep Neural Networks Triggered by Fault Injection Attack on Image Sensor Interface.

A backdoor attack is a type of attack method that induces deep neural network (DNN) misclassification. The adversary who aims to trigger the backdoor attack inputs the image with a specific pattern (the adversarial mark) into the DNN model (backdoor model). In general, the adversary mark is created on the physical object input to an image by capturing a photo. With this conventional method, the success of the backdoor attack is not stable because the size and position change depending on the shooting environment. So far, we have proposed a method of creating an adversarial mark for triggering backdoor attacks by means of a fault injection attack on the mobile industry processor interface (MIPI), which is the image sensor interface. We propose the image tampering model, with which the adversarial mark can be generated in the actual fault injection to create the adversarial mark pattern. Then, the backdoor model was trained with poison data images, which the proposed simulation model created. We conducted a backdoor attack experiment using a backdoor model trained on a dataset containing 5% poison data. The clean data accuracy in normal operation was 91%; nevertheless, the attack success rate with fault injection was 83%.

Modulation of phytotoxic and cytogenetic effects of cadmium by humic acid: Findings from a short-term plant-based bioassay.

The study of the modulation of the toxicity of heavy metals by coexisting chemicals in the environment is vital for realistic ecological risk assessment. Our study was aimed at determining possible toxicity modulations of Cd by humic acid (HA) using the Allium cepa test system. A. cepa bulbs were exposed to Cd (1 and 5 mg/L) and HA (10 mg/L) individually or in mixtures. The root lengths of the bulbs and cytogenetic endpoints in root meristematic cells, including the mitotic index (MI), nuclear abnormalities (NAs), and chromosomal abnormalities (CAs), were determined. The results revealed that the MIs of A. cepa co-exposed to HA and Cd were significantly recovered by >15% compared with those of A. cepa subjected to Cd-only treatments, and this response was more sensitive than the phytotoxic response (root length). Furthermore, the burden of NAs was significantly decreased in the co-exposed bulbs by >20% compared with bulbs with Cd-only treatments. The frequencies of CAs were also reduced in the bulbs co-exposed to HA and 1 and 5 mg/L Cd by >15 and >25%, respectively, compared with bulbs receiving Cd-only treatments. Therefore, our findings indicated that HA plays a significant protective role in Cd toxicity in A. cepa.

Egeria densa organic extracts: an eco-friendly approach to suppress Microcystis aeruginosa growth through allelopathy.

Macrophytes play a significant role in shaping plankton communities by shading, controlling water turbulence, and nutrient availability, while also producing allelochemicals causing varying effects on different organisms. Many researchers have shown that when live macrophytes are present, they inhibit cyanobacteria. However, their widespread use is often limited due to numerous concerns, including invasive characteristics. This study focused on the applicability of Egeria densa extracts to suppress Microcystis aeruginosa. We employed pure water and dimethyl sulfoxide, to obtain compounds from E. densa. The results revealed that E. densa aqueous extracts stimulated M. aeruginosa growth, whereas organic extracts exhibited suppression. Specifically, at low concentrations of organics extracts (0.5 and 1 g/L), after day 4, the growth inhibition was confirmed by significantly higher (p < 0.05) stress levels shown in cells treated with low concentrations. The highest inhibition rate of 32% was observed at 0.5 g/L. However, high concentrations of organic extracts (3 and 6 g/L), showed increased growth compared with control. These results suggest that high concentrations of organic extracts from E. densa potentially suppress allelochemical-induced M. aeruginosa inhibition due to high nutrient availability. In comparison with an aqueous solvent, the use of organic solvent seems to be more effective in efficiently extracting allelochemicals from E. densa.

CHIP-associated mutant ASXL1 in blood cells promotes solid tumor progression.

Clonal hematopoiesis of indeterminate potential (CHIP) is an age-associated phenomenon characterized by clonal expansion of blood cells harboring somatic mutations in hematopoietic genes, including DNMT3A, TET2, and ASXL1. Clinical evidence suggests that CHIP is highly prevalent and associated with poor prognosis in solid-tumor patients. However, whether blood cells with CHIP mutations play a causal role in promoting the development of solid tumors remained unclear. Using conditional knock-in mice that express CHIP-associated mutant Asxl1 (Asxl1-MT), we showed that expression of Asxl1-MT in T cells, but not in myeloid cells, promoted solid-tumor progression in syngeneic transplantation models. We also demonstrated that Asxl1-MT-expressing blood cells accelerated the development of spontaneous mammary tumors induced by MMTV-PyMT. Intratumor analysis of the mammary tumors revealed the reduced T-cell infiltration at tumor sites and programmed death receptor-1 (PD-1) upregulation in CD8+ T cells in MMTV-PyMT/Asxl1-MT mice. In addition, we found that Asxl1-MT induced T-cell dysregulation, including aberrant intrathymic T-cell development, decreased CD4/CD8 ratio, and naïve-memory imbalance in peripheral T cells. These results indicate that Asxl1-MT perturbs T-cell development and function, which contributes to creating a protumor microenvironment for solid tumors. Thus, our findings raise the possibility that ASXL1-mutated blood cells exacerbate solid-tumor progression in ASXL1-CHIP carriers.

High-Affinity Ratiometric Fluorescence Probe Based on 6-Amino-2,2'-Bipyridine Scaffold for Endogenous Zn2+ and Its Application to Living Cells.

Zinc is an essential trace element involved in many biological activities; however, its functions are not fully understood. To elucidate the role of endogenous labile Zn2+, we developed a novel ratiometric fluorescence probe, 5-(4-methoxyphenyl)-4-(methylsulfanyl)-[2,2'-bipyridin]-6-amine (6 (rBpyZ)) based on the 6-amino-2,2'-bipyridine scaffold, which acts as both the chelating agent for Zn2+ and the fluorescent moiety. The methoxy group acted as an electron donor, enabling the intramolecular charge transfer state of 6 (rBpyZ), and a ratiometric fluorescence response consisting of a decrease at the emission wavelength of 438 nm and a corresponding increase at the emission wavelength of 465 nm was observed. The ratiometric probe 6 (rBpyZ) exhibited a nanomolar-level dissociation constant (Kd = 0.77 nM), a large Stokes shift (139 nm), and an excellent detection limit (0.10 nM) under physiological conditions. Moreover, fluorescence imaging using A549 human lung adenocarcinoma cells revealed that 6 (rBpyZ) had good cell membrane permeability and could clearly visualize endogenous labile Zn2+. These results suggest that the ratiometric fluorescence probe 6 (rBpyZ) has considerable potential as a valuable tool for understanding the role of Zn2+ in living systems.

[Pathophysiology of hematological malignancies associated with ASXL1 mutations].

Somatic mutations in the epigenetic regulator ASXL1 are considered a poor prognostic factor in myeloid malignancies, including myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). ASXL1 mutations coexist with other mutations in majority of patients, suggesting that its mutation alone is insufficient to cause cancer. ASXL1 mutations have been detected in age-related clonal hematopoiesis (CH), which has been linked to an increased risk of hematological malignancies. Therefore, ASXL1 mutations are likely to be one of the first events in the tumorigenesis process. With our most recent findings, we summarize the mechanisms by which ASXL1 mutations cause CH and hematological malignancies in this review.

Selective Cadmium Fluorescence Probe Based on Bis-heterocyclic Molecule and its Imaging in Cells.

Fluorescence probes that selectively image cadmium are useful for detecting and tracking the amount of Cd2+ in cells and tissues. In this study, we designed and synthesized a novel Cd2+ fluorescence probe based on the pyridine-pyrimidine structure, 4-(methylsulfanyl)-6-(pyridin-2-yl)pyrimidin-2-amine (3), as a low-molecular-weight fluorescence probe for Cd2+. Compound 3 could successfully discriminate between Cd2+ and Zn2+ and exhibited a highly selective turn-on response toward Cd2+ over biologically related metal ions. The dissociation constant (Kd) and the limit of detection (LOD) of 5.4 × 10- 6 mol L- 1 and 4.4 × 10- 7 mol L- 1, respectively, were calculated using fluorescence titration experiments. Studies with closely related analogs showed that the bis-heterocyclic moiety of 3 acted as both a coordination site for Cd2+ and a fluorophore. Further, the methylsulfanyl group of compound 3 is essential for achieving selective and sensitive Cd2+ detection. Fluorescence microscopy studies using living cells revealed that the cell membrane permeability of compound 3 is sufficient to detect intracellular Cd2+. These results indicate that novel bis-heterocyclic molecule 3 has considerable potential as a fluorescence probe for Cd2+ in biological applications.

Area-Efficient Post-Processing Circuits for Physically Unclonable Function with 2-Mpixel CMOS Image Sensor.

In order to realize image information security starting from the data source, challenge-response (CR) device authentication, based on a Physically Unclonable Function (PUF) with a 2 Mpixel CMOS image sensor (CIS), is studied, in which variation of the transistor in the pixel array is utilized. As each CR pair can be used only once to make the CIS PUF resistant to the modeling attack, CR authentication with CIS can be carried out 4050 times, with basic post-processing to generate the PUF ID. If a larger number of authentications is required, advanced post-processing using Lehmer encoding can be utilized to carry out authentication 14,858 times. According to the PUF performance evaluation, the authentication error rate is less than 0.001 ppm. Furthermore, the area overhead of the CIS chip for the basic and advanced post-processing is only 1% and 2%, respectively, based on a Verilog HDL model circuit design.

Assessment of the applicability of a low-cost sensor-based methane monitoring system for continuous multi-channel sampling.

Systems that are made of several low-cost gas sensors with automatic gas sampling may have the potential to serve as reliable fast methane analyzers. However, there is a lack of reports about such types of systems evaluated under field conditions. Here, we developed a continuous methane monitoring system with automated gas sampling unit using low-cost gas sensors, TGS 2611 and MQ-4, that use a simple cloud-based data acquisition platform. We verified the consistency, repeatability, and reproducibility of the data obtained by TGS 2611 and MQ-4 low-cost gas sensors by measuring high- and low-concentration methane samples. The normalized root-mean-square errors (NRMSEs) of the samples with high methane concentrations, [CH4] of 3, 4, 6, and 7%, were 0.0788, 0.0696, 0.1198, and 0.0719 for the TGS 2611 sensor, respectively, and were confirmed using a gas chromatograph as a reference analyzer. The NRMSEs of the samples with low [CH4] of 0.096, 0.145, 0.193, and 0.241% measured by the TGS 2611 sensor were 0.0641, 0.1749, 0.0157, and 0.1613, whereas those NRMSEs of the same concentrations measured by the MQ-4 sensor were 0.3143, 0.5766, 0.6301, and 0.6859, respectively. Laboratory-scale anaerobic digesters were tested using the developed system. The anaerobic digesters were continuously operated for 2 months, demonstrating the potential use of sensors for detecting and monitoring methane in the field level application. This study utilized a unique way to combine the advantages of low-cost sensors and develop a reliable monitoring system by minimizing drawbacks of low-cost sensors.

The role of ASXL1 in hematopoiesis and myeloid malignancies.

Recent high-throughput genome-wide sequencing studies have identified recurrent somatic mutations in myeloid neoplasms. An epigenetic regulator, Additional sex combs-like 1 (ASXL1), is one of the most frequently mutated genes in all subtypes of myeloid malignancies. ASXL1 mutations are also frequently detected in clonal hematopoiesis, which is associated with an increased risk of mortality. Therefore, it is important to understand how ASXL1 mutations contribute to clonal expansion and myeloid transformation in hematopoietic cells. Studies using ASXL1-depleted human hematopoietic cells and Asxl1 knockout mice have shown that deletion of wild-type ASXL1 protein leads to impaired hematopoiesis and accelerates myeloid malignancies via loss of interaction with polycomb repressive complex 2 proteins. On the other hand, ASXL1 mutations in myeloid neoplasms typically occur near the last exon and result in the expression of C-terminally truncated mutant ASXL1 protein. Biological studies and biochemical analyses of this variant have shed light on its dominant-negative and gain-of-function features in myeloid transformation via a variety of epigenetic changes. Based on these results, it would be possible to establish novel promising therapeutic strategies for myeloid malignancies harboring ASXL1 mutations by blocking interactions between ASXL1 and associating epigenetic regulators. Here, we summarize the clinical implications of ASXL1 mutations, the role of wild-type ASXL1 in normal hematopoiesis, and oncogenic functions of mutant ASXL1 in myeloid neoplasms.

Behavior of bentonite in an aqueous electrolytic solution - evaluation of electrolytic aggregation for adsorption capacity of Cd2+ ions onto bentonite.

In this study, we used aqueous solutions containing 1 mg/L of Cd2+ for electrolysis while varying the current density (CD), amount of bentonite added and the effective submerged area to investigate the adsorption capacity of Cd2+ ions onto bentonite by electrolytic aggregation. The adsorption of Cd2+ ions increased with increasing amount of bentonite added to the electrolytic solution. The addition of bentonite also regulated the pH of the electrolytic solution during the electrolysis process in addition to the hydrolysis of water. The maximum adsorption capacities at equilibrium (qe) for current densities of 3.14 and 7.49 mA/cm2 (i.e. for 2 and 1 L electrolytic solutions) with 0.2 g of bentonite were 4.54 and 2.92 mg/g, respectively. The removal of Cd2+ (RCd) clearly depended on the pH of the electrolytic solution. Moreover, qe decreased with increasing amount of bentonite used for electrolytic aggregation. The findings of this study will be useful for understanding the aggregation of clay particles under electrolysis and their adsorption behaviors.

Virus-induced gene silencing (VIGS)-mediated functional characterization of two genes involved in lignocellulosic secondary cell wall formation.

KEY MESSAGE: Functional characterization of two tobacco genes, one involved in xylan synthesis and the other, a positive regulator of secondary cell wall formation, is reported. Lignocellulosic secondary cell walls (SCW) provide essential plant materials for the production of second-generation bioethanol. Therefore, thorough understanding of the process of SCW formation in plants is beneficial for efficient bioethanol production. Recently, we provided the first proof-of-concept for using virus-induced gene silencing (VIGS) approach for rapid functional characterization of nine genes involved in cellulose, hemicellulose and lignin synthesis during SCW formation. Here, we report VIGS-mediated functional characterization of two tobacco genes involved in SCW formation. Stems of VIGS plants silenced for both selected genes showed increased amount of xylem formation but thinner cell walls than controls. These results were further confirmed by production of stable transgenic tobacco plants manipulated in expression of these genes. Stems of stable transgenic tobacco plants silenced for these two genes showed increased xylem proliferation with thinner walls, whereas transgenic tobacco plants overexpressing these two genes showed increased fiber cell wall thickness but no change in xylem proliferation. These two selected genes were later identified as possible members of DUF579 family involved in xylan synthesis and KNAT7 transcription factor family involved in positive regulation of SCW formation, respectively. Glycome analyses of cell walls showed increased polysaccharide extractability in 1 M KOH extracts of both VIGS-NbDUF579 and VIGS-NbKNAT7 lines suggestive of cell wall loosening. Also, VIGS-NbDUF579 and VIGS-NbKNAT7 lines showed increased saccharification rates (74.5 and 40 % higher than controls, respectively). All these properties are highly desirable for producing higher quantities of bioethanol from lignocellulosic materials of bioenergy plants.

Decreased number of Gemini of coiled bodies and U12 snRNA level in amyotrophic lateral sclerosis.

Disappearance of TAR-DNA-binding protein 43 kDa (TDP-43) from the nucleus contributes to the pathogenesis of amyotrophic lateral sclerosis (ALS), but the nuclear function of TDP-43 is not yet fully understood. TDP-43 associates with nuclear bodies including Gemini of coiled bodies (GEMs). GEMs contribute to the biogenesis of uridine-rich small nuclear RNA (U snRNA), a component of splicing machinery. The number of GEMs and a subset of U snRNAs decrease in spinal muscular atrophy, a lower motor neuron disease, suggesting that alteration of U snRNAs may also underlie the molecular pathogenesis of ALS. Here, we investigated the number of GEMs and U11/12-type small nuclear ribonucleoproteins (snRNP) by immunohistochemistry and the level of U snRNAs using real-time quantitative RT-PCR in ALS tissues. GEMs decreased in both TDP-43-depleted HeLa cells and spinal motor neurons in ALS patients. Levels of several U snRNAs decreased in TDP-43-depleted SH-SY5Y and U87-MG cells. The level of U12 snRNA was decreased in tissues affected by ALS (spinal cord, motor cortex and thalamus) but not in tissues unaffected by ALS (cerebellum, kidney and muscle). Immunohistochemical analysis revealed the decrease in U11/12-type snRNP in spinal motor neurons of ALS patients. These findings suggest that loss of TDP-43 function decreases the number of GEMs, which is followed by a disturbance of pre-mRNA splicing by the U11/U12 spliceosome in tissues affected by ALS.

First clinical experience of the looped Inoue balloon technique for antegrade percutaneous balloon aortic valvuloplasty.

Balloon aortic valvuloplasty (BAV) has played a limited role in the management of patients with severe aortic stenosis. However, BAV is being performed more frequently these days with the emergence of transcatheter aortic valve implantation (TAVI). We previously described a technique named "looped Inoue balloon technique" to simplify the antegrade transvenous BAV by making a loop in the left atrium using two stylets. We present a case in which the looped Inoue balloon technique was successfully applied. The patient was an 83-year-old woman with progressive dyspnea due to severe aortic stenosis. The aortic valve area was 0.39 cm(2) with a mean transvalvular gradient of 46 mmHg. The patient was deemed high risk for surgical aortic valve replacement or TAVI in view of the multiple comorbidities and frailty. Antegrade BAV using the looped Inoue balloon technique was performed. The procedure was successful without any complications. The post procedural aortic valve area increased to 1.15 cm(2) with a mean pressure gradient of 23 mmHg. This is the first report of clinical use of the looped Inoue balloon technique for antegrade BAV.

Enhanced accumulation of fatty acids and triacylglycerols in transgenic tobacco stems for enhanced bioenergy production.

KEY MESSAGE: We report a novel approach for enhanced accumulation of fatty acids and triacylglycerols for utilization as biodiesel in transgenic tobacco stems through xylem-specific expression of Arabidopsis DGAT1 and LEC2 genes. The use of plant biomass for production of bioethanol and biodiesel has an enormous potential to revolutionize the global bioenergy outlook. Several studies have recently been initiated to genetically engineer oil production in seeds of crop plants to improve biodiesel production. However, the "food versus fuel" issues have also sparked some studies for enhanced accumulation of oils in vegetative tissues like leaves. But in the case of bioenergy crops, use of woody stems is more practical than leaves. Here, we report the enhanced accumulation of fatty acids (FAs) and triacylglycerols (TAGs) in stems of transgenic tobacco plants expressing Arabidopsis diacylglycerol acyltransferase 1 (DGAT1) and leafy cotyledon2 (LEC2) genes under a developing xylem-specific cellulose synthase promoter from aspen trees. The transgenic tobacco plants accumulated significantly higher amounts of FAs in their stems. On an average, DGAT1 and LEC2 overexpression showed a 63 and 80% increase in total FA production in mature stems of transgenic plants over that of controls, respectively. In addition, selected DGAT1 and LEC2 overexpression lines showed enhanced levels of TAGs in stems with higher accumulation of 16:0, 18:2 and 18:3 TAGs. In LEC2 lines, the relative mRNA levels of the downstream genes encoding plastidic proteins involved in FA synthesis and accumulation were also elevated. Thus, here, we provide a proof of concept for our approach of enhancing total energy yield per plant through accumulation of higher levels of FAs in transgenic stems for biodiesel production.

Cytogenotoxicity of raw and treated dairy manure slurry by two-stage chemical and electrocoagulation: An application of the Allium cepa bioassay.

Livestock farming is an essential agricultural practice. However, the improper management of livestock wastes and discharge of untreated or partially treated livestock manure slurry poses significant environmental problems. In this study, we aimed to compare the cytogenotoxic potential of untreated and treated dairy manure slurry treated with a two-stage chemical and electrocoagulation (EC) using the Allium cepa bioassay. The A. cepa bioassay is a well-established standard tool for assessing the cytogenotoxic effects of environmental contaminants, especially those that are occurred as complex contaminant mixtures. The dairy manure slurry was subjected to chemical treatment utilizing polyaluminum chloride (PAC) and cationic polyacrylamide (CPAM) at optimized conditions, followed by EC utilizing either aluminum (Al) or steel anodes. The treated and untreated samples were then evaluated for their potential cytogenotoxicty using the A. cepa bioassay, by measuring the nuclear abnormalities (NAs) and chromosomal aberrations (CAs), along with the mitotic indices (MIs). Our findings revealed a significant reduction in cytogenotoxic indicators in the treated liquid fraction compared to the untreated dairy manure slurry. Specifically, the frequency of total NAs showed a significant reduction from 154 ‰ to 37 ‰ when the dairy manure slurry was treated with chemical coagulation followed by EC utilizing an Al anode. Moreover, the MI exhibited a significant improvement from 7 ‰ to 123 ‰, suggesting the mitigation of toxic effects. These results collectively demonstrate the effectiveness of the two-stage chemical and EC treatment under optimal conditions in treating diary manure slurry while reducing its cytogenotoxicity for living systems. The A. cepa bioassay proved to be a sensitive and reliable method for assessing the toxicity of the treated samples. The efficient solid-liquid separation and the reduction of toxicity in the liquid fraction for biological systems achieved through this treatment process highlight its potential for sustainable management of livestock waste and the preservation of water quality. Nevertheless, further studies are required to assess the toxicity of solid fraction.

Efficient selection of a biallelic and nonchimeric gene-edited tree using Oxford Nanopore Technologies sequencing.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease system is a versatile and essential biotechnological tool in the life sciences that allows efficient genome editing. When generating gene-edited trees, T0-generation plants are often used for subsequent analysis because of the time that is required to obtain the desired mutants via crossing. However, T0-generation plants exhibit various unexpected mutations, which emphasizes the need to identify mutants with expected mutation patterns. The two critical checkpoints in this process are to confirm the expected mutation patterns in both alleles and to exclude somatic chimeric plants. In this study, we generated gene-edited Cryptomeria japonica plants and established a method to determine chimerism and mutation patterns using fragment analysis and Oxford Nanopore Technologies (ONT)-based amplicon sequencing. In the first screening, fragment analysis, i.e., indel detection via amplicon analysis, was used to predict indel mutation patterns in both alleles and to discriminate somatic chimeric plants in 188 candidate mutants. In the second screening, we precisely determined the mutation patterns and chimerism in the mutants using ONT-based amplicon sequencing, where confirmation of both alleles can be achieved using allele-specific markers flanking the single guide RNA target site. In the present study, a bioinformatic analysis procedure was developed and provided for the rapid and accurate determination of DNA mutation patterns using ONT-based amplicon sequencing. As ONT amplicon sequencing has a low running cost compared with other long-read analysis methods, such as PacBio, it is a powerful tool in plant genetics and biotechnology to select gene-edited plants with expected indel patterns in the T0-generation.