Surface engineering of biochar toward simultaneously generating superamphiphilicity and catalytic activity for strengthening Pickering interfacial catalysis.

Multifunctional carbon materials have revealed distinctive features and excellent performance in the field of catalysis. However, the facile fabrication of bifunctional carbon materials with special wettability and catalytic activity remains a grand challenge in Pickering emulsion catalysis. Herein, we reported one-step construction of bifunctional biochar with superamphiphilicity and catalytic activity directly from the thermolysis of sawdust and 1-butyl-3-methylimidazolium tetrafluoroborate for enhancing the oxidation of benzyl alcohol in Pickering emulsion. Co-doping of B and F enhanced the hydrophilicity of biochar, and the oleophilicity of biochar was kept simultaneously. Conversion became 4 times using bifunctional biochar compared with blank results during the oxidation of benzyl alcohol. More interestingly, the turnover frequency (TOF) value using bifunctional biochar enhanced 61% than that employing N-doped superamphiphilic carbon without catalytic activity. Catalytic activities of bifunctional biochar could be ascribed to the existence of different chemical bonds containing the element B. This work paves a path toward rational design of bifunctional biochar materials with special wettability and catalytic activity for greatly enhancing the liquid-liquid biphasic reaction efficiencies.

Epidermolysis Bullosa: Two rare case reports of COL7A1 and EBS-GEN SEV KRT14 variants with review of literature.

EPIDERMOLYSIS: Bullosa is a rare hereditary skin condition that causes blisters. Genes encoding structural proteins at or near the dermal-epidermal junction are mutated recessively or dominantly, and this is the primary cause of EB. Herein, two Chinese boys were diagnosed with the condition, each with a different variant in a gene that serves as a reference for EB genetic counseling. Skincare significantly impacted their prognosis and quality of life. CASE PRESENTATION: Two Chinese boys, with phenotypically normal parents, have been diagnosed with distinct blister symptoms, one with Dominant Dystrophic Epidermolysis Bullosa and the other with a severe form of Epidermolysis Bullosa Simplex. The first patient had a G-to-A variant in the COL7A1 allele, at nucleotide position 6163 which was named "G2055A". The proband is heterozygous for Dystrophic Epidermolysis Bullosa due to a COL7A1 allele with a glycine substitution at the triple helix domain. A similar variant has been discovered in his mother, indicating its potential transmission to future generations. Another patient had severe Epidermolysis Bullosa Simplex with a rare c.377T > A  variant resulting in substitution of amino acid p.Leu126Arg (NM_000526.5 (c.377T > G, p.Leu126Arg) in the Keratin 14 gene. In prior literature, Keratin 14 has been associated with an excellent prognosis. However, our patient with this infrequent variant tragically died from sepsis at 21 days old. There has been a reported occurrence of the variant only once. CONCLUSION: Our study reveals that Epidermolysis Bullosa patients with COL7A1 c.6163G > A and KRT14 c.377T>A variants have different clinical presentations, with dominant forms of Dystrophic EB having milder phenotypes than recessive ones. Thus, the better prognosis in the c.6163G > A patient. Furthermore, c.377T>A patient was more prone to infection than the patient with c.6163G>A gene variant. Genetic testing is crucial for identifying the specific variant responsible and improving treatment options.

Molecular Insights into the Dual Promotion-Inhibition Effects of NaCl at Various Concentrations on the CO2 Hydrate Growth: A Molecular Simulation Study.

Hydrate-based CO2 storage in the ocean is considered a potential method for mitigating the greenhouse effect. Numerous studies demonstrated that NaCl exhibited the dual effects of promotion and inhibition in the nucleation and growth processes of CO2 hydrate, whose mechanisms remain unclear. In this study, the effects of NaCl at various concentrations on the CO2 hydrate growth and crystal are investigated. The independent gradient model based on Hirshfeld partition, electrostatic potential, and binding energy is conducted to study the interaction between ions and water molecules. The motion trajectories of ions are observed at the molecular level to reflect the impact of ion motion on hydrate growth. The results show that the influence of NaCl on hydrate growth depends on a delicate balance of dual promotion-inhibition effects. NaCl can combine more water molecules and provide a transport channel of CO2 to promote hydrate growth at low concentrations. Meanwhile, the promoting effects shift toward inhibition with increasing NaCl concentrations. In a word, this paper proposes a novel mechanism for the dual promotion-inhibition effects of NaCl on hydrate growth, which is significant for further research on hydrate-based CO2 storage in the ocean.

Encapsulated allyl isothiocyanate improves soil distribution, efficacy against soil-borne pathogens and tomato yield.

BACKGROUND: Crop quality, yield and farmer income are reduced by soil-borne diseases, nematodes and weeds, although these can be controlled by allyl isothiocyanate (AITC), a plant-derived soil fumigant. However, its efficacy against soil-borne pathogens varies, mainly because of its chemical instability and uneven distribution in the soil. Formulation modification is an effective way to optimize pesticide application. We encapsulated AITC in modified diatomite granules (GR) and measured the formulation's loading content and stability, environmental fate and efficacy against soil-borne pathogens, and its impact on the growth and yield of tomatoes. RESULTS: We observed that an AITC loading content in the granules of 27.6% resulted in a degradation half-life of GR that was 1.94 times longer than 20% AITC emulsifiable concentrate in water (EW) and shorter than AITC technical (TC) grade. The stable and more even distribution of GR in soil resulted in relatively consistent and acceptable control of soil-borne pathogens. Soil containing AITC residues that remained 10-24 days after GR fumigation were not phytotoxic to cucumber seeds. GR significantly reduced soil-borne pest populations, and tomato growth and yield increased as AITC dosage increased. GR containing an AITC dose of 20 g m-2 effectively controlled pathogens in soil for about 7 months and improved tomato yield by 108%. CONCLUSION: Our research demonstrates the benefits of soil fumigation with loaded AITC over other formulations for effective pest control, and improved tomato plant growth and fruit yield. Fumigant encapsulation appears to be a useful method to improve pest and disease control, environmental performance and fumigant commercial sustainability. © 2024 Society of Chemical Industry.

The potential for reducing aflatoxin B1 contamination of stored peanuts by soil disinfection.

Aflatoxins from the fungus Aspergillus flavus (A. flavus) that contaminate stored peanuts is a major hazard to human health worldwide. Reducing A. flavus in soil can decrease the risk of aflatoxins in stored peanuts. In this experiment, we determined whether peanuts grown on soil fumigated with dazomet (DZ), metham sodium (MS), allyl isothiocyanate (AITC), chloropicrin (PIC) or dimethyl disulfide (DMDS) would reduce of the quantity of A. flavus and its toxin's presence. The results of bioassays and field tests showed that PIC was the most effective fumigant for preventing and controlling A. flavus, followed by MS. PIC and MS applied to the soil for 14 d resulted in LD50 values against A. flavus of 3.558 and 4.893 mg kg-1, respectively, leading to almost 100% and 98.82% effectiveness of A. flavus, respectively. Peanuts harvested from fumigated soil and then stored for 60 d resulted in undetectable levels of aflatoxin B1 (AFB1) compared to unfumigated soil that contained 0.64 ug kg-1 of AFB1, which suggested that soil fumigation can reduce the probability of aflatoxin contamination during peanut storage and showed the potential to increase the safety of peanuts consumed by humans. Further research is planned to determine the practical value of our research in commercial practice.

The effect of ghrelin on bursa and cecal tonsils of chickens infected with an attenuated virus strain of infectious bursal disease virus.

Infectious bursal disease (IBD) significantly affects the poultry industry, causing substantial economic losses. This study aimed to investigate the effects of ghrelin on chicks infected with an attenuated virus strain of IBDV (aIBDV). Chicks were divided into 3 groups: a control group (group I), an aIBDV infection group (group II), and a ghrelin + aIBDV infection group (group III). Mice in groups II and III were fed until they reached 19 d of age and then inoculated with aIBDV to establish a subclinical infection model. Group III received an intraperitoneal injection of 0.5 nmol/100 g ghrelin from d 17 to 23. The present study utilized paraffin sectioning, H&E staining, and immunohistochemical staining to examine the effects of ghrelin on the bursa of fabricius and cecum tonsils in aIBDV-infected chicks. The results indicated that at 3 d postinfection (dpi), the average body weight of group III was significantly greater than that of group II (P < 0.05). At 3 and 7 dpi, the proportion of large lymphoid follicles in the bursa of fabricius in group III was notably greater than that in group II (P < 0.05). aIBDV infection resulted in bleeding, edema, and fibrosis in the cecal mucosal layer of chicks, but ghrelin administration mitigated these pathological changes. At 3 and 7 dpi, the thickness of the lamina propria in the cecal tonsils of group III was significantly lower than that in the cecal tonsils of group II (P < 0.05). Additionally, the percentage of large lymphoid follicles in the cecal tonsils of group III was significantly greater than that in group II at 3 and 5 dpi (P < 0.05). There were significantly fewer macrophages in the cecal tonsils of group III than in those of group II at 1, 3, and 5 dpi (P < 0.05). In conclusion, ghrelin supplementation improved performance and mitigated bursal atrophy in aIBDV-infected chicks. It also reduced histological lesions and immune responses in the cecum tonsil. Notably, the reduction in macrophages in the cecum tonsil following ghrelin administration may decrease the risk of aIBDV spread.

Trichoderma spp. promotes ginseng biomass by influencing the soil microbial community.

Introduction: Ginseng (Panax ginseng C.A. Meyer) has multiple effects on human health; however, soil degradation seriously affects its yield. Trichoderma spp. play an important role in improving plant biomass by influencing the soil environment. Therefore, it is necessary to screen efficient Trichoderma strains that can increase ginseng biomass and determine their mechanisms. Methods: Herein, we selected six Trichoderma species (T. brevicompactum, T. velutinum, T. viridescens, T. atroviride, T. koningiopsis, and T. saturnisporum) isolated from ginseng rhizosphere soil, and evaluated their growth promoting effects on ginseng and their influence on the microbiome and chemical attributes of the ginseng rhizosphere soil. Results: Except for T. saturnisporum (F), compared with the control, the other five species increased ginseng biomass. In terms of chemical properties, the pH value, available potassium content, and available phosphorus content in the ginseng rhizosphere soil increased by 1.16-5.85%, 0.16-14.03%, and 3.92-38.64%, respectively, after root irrigation with spores of Trichoderma species. For the soil microbiome, fungal Chao1 and Ace richness indices decreased. Application of Trichoderma enhanced the relative level of Proteobacteria, but reduced the relative level of Ascomycota. At the genus level, application of Trichoderma enhanced the relative levels of Sphingomonas, Blastomonas, and Trichoderma, but reduced the relative level of Fusarium. Available K and available P were the most important elements that affected the structure of the bacterial community, while total K was the most influential element for the structure of the fungal community structure. Conclusion: The results indicated that the application of Trichoderma spp. could increase soil nutrients and regulate the structure and composition of the soil microbial community, thereby enhancing the biomass of ginseng. The results will provide guidance for soil improvement in ginseng cultivation.

Fumigation alters the manganese-oxidizing microbial communities to enhance soil manganese availability and increase tomato yield.

Manganese is one of the essential trace elements for plants to maintain normal life activities. Soil fumigation, while effectively controlling soil-borne diseases, can also improve the cycling of soil nutrient elements. MiSeq amplicon sequencing is used to determine the composition of soil microbial communities, and structural equation modeling and the random forest algorithm are employed to conduct a correlation analysis between key manganese-oxidizing microorganisms and soil manganese availability. This experiment investigated the microbial mechanisms behind the observed increase in available manganese in soil after fumigation. The key findings revealed that Bacillus, GeoBacillus, GraciliBacillus, Chungangia, and Pseudoxanthomonas play crucial roles in influencing the variation in soil available manganese content. Fumigation was found to elevate the abundance of Bacillus. Moreover, laccase activity emerged as another significant factor impacting soil manganese availability, showing an indirect correlation with available manganese content and contributing to 58 % of the observed variation in available manganese content. In summary, alterations in the communities of manganese-oxidizing microorganisms following soil fumigation are pivotal for enhancing soil manganese availability.

Effects of dietary supplementation of fermented Artemisia argyi on growth performance, slaughter performance, and meat quality in broilers.

Artemisia argyi (AA) is promising as a potential feed additive. Microbial fermentation is beneficial to the degradation of cell walls and the better release of bioactive compounds of AA. However, there are few reports on the application of fermented AA as a feed additive for broilers. The present study intended to evaluate the application value of fermented AA as a feed additive for broilers by examining the effects of the dietary supplementation of Aspergillus niger-fermented AA and unfermented AA on growth performance, slaughter performance, and meat quality of brokers. A total of 360 newly hatched (1-day-old) broilers with similar body weight were randomly divided into the following 5 groups: basal diet group as control (C) group, basal diet +3% unfermented AA (E1) group, basal diet + 1% fermented AA (E2) group, basal diet + 3% fermented AA (E3) group, basal diet + 5% fermented AA (E4) group. Each group included 6 replicates with 12 broilers per replicate, and the feeding trail lasted for 48 d. Body weight and feed intake were recorded every 2 wk, and the feed gain ratio was calculated to assess growth performance. At 42 d, 6 broilers from each group were slaughtered, and the carcass traits were calculated. The results showed that compared with the control group, Aspergillus Niger could effectively destroy AA fiber, which contributed to better release of AA bioactive compounds. Moreover, dietary supplementation with AA could improve the growth performance of broilers (P < 0.05), and the effect of fermented AA was better than unfermented AA, especially 3% fermented AA. From 28 to 42 d, compared with the control group, the average daily gain of broilers in the group supplementation with 3% fermented AA was significantly increased (P < 0.05), and the feed-to-gain ratio was decreased (P < 0.05). At 42 d, the dressing percentage, half-eviscerated carcass percentage, eviscerated carcass percentage, and breast muscle percentage of broilers in the groups of 1, 3, and 5% fermented AA diets were significantly improved (P < 0.05), and the thigh muscle percentage of broilers in the group with 3% fermented AA diets was significantly improved (P < 0.05). Meanwhile, the meat quality of broilers in the group with fermented AA diets was also significantly improved. Birds in AA groups had higher a* value and lower shear force of breast muscle, especially the group supplementation with 3% fermented AA (P < 0.05). In conclusion, fermented AA has good application value as a potential feed additive for broilers, dietary supplementation of fermented AA can improve the production performance and meat quality of broiler chickens, of which 3% fermented AA is more effective.

An applicability study of rapid artificial intelligence-assisted compressed sensing (ACS) in anal fistula magnetic resonance imaging.

Objective: To evaluate the applicability of artificial intelligence-assisted compressed sensing (ACS) to anal fistula magnetic resonance imaging (MRI). Methods: 51 patients were included in this study and underwent T2-weighted sequence of MRI examinations both with ACS and without ACS technology in a 3.0 T MR scanner. Subjective image quality scores, and objective image quality-related metrics including scanning time, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR), were evaluated and statistically compared between the images collected with and without ACS. Results: No significant difference in the subjective image quality of lesion conspicuity was observed between the two groups. However, ACS MRI decreased the acquisition time with regard to control group (74.00 s vs. 156.00 s). Besides, SNR of perianal and muscle in the ACS group was significantly higher than that of the control group (164.07 ± 33.35 vs 130.81 ± 29.10, p < 0.001; 109.87 ± 22.01 vs 87.61 ± 17.95, p < 0.001; respectively). The CNR was significantly higher in the ACS group than in the control group (54.02 ± 23.98 vs 43.20 ± 21.00; p < 0.001). Moreover, the accuracy rate of the ACS groups in evaluating the direction and internal opening of the fistula was 88.89 %, exactly the same as that of the control group. Conclusion: We demonstrated the applicability of using ACS to accelerate MR of anal fistulas with improved SNR and CNR. Meanwhile, the accuracy rates of the ACS group and the control were equivalent in evaluating the direction and internal opening of the fistula, based on the results of surgical exploration.

Correction to "Microstructure of Heavy Oil Components and Mechanism of Influence on Viscosity of Heavy Oil".

[This corrects the article DOI: 10.1021/acsomega.2c07713.].

ADP-ribosylation factor 6 promotes infectious bursal disease virus replication by affecting the internalization process via clathrin.

ADP-ribosylation factor 6 (ARF6) is a small G protein with extensive functions, including regulation of cellular membrane transport and viral infection. Infectious bursal disease (IBD) is caused by infectious bursal disease virus (IBDV), which mainly invades the bursa of Fabricius and causes low immunity in poultry. Our study demonstrated that IBDV infection could promote the expression of ARF6; however, the underlying mechanism remains unclear. Herein, the function of ARF6 in IBDV infection was explored, and it was revealed that viral replication was significantly promoted by ARF6 overexpression and hampered by siRNA-mediated inhibition of ARF6. Using two site mutants of ARF6 (ARF6-T27N and ARF6-Q67L), we found that IBDV replication was repressed by ARF6-T27N, indicating that ARF6 promotes IBDV replication. Further exploration of its mechanism revealed that ARF6 affects the copy number of IBDVs entering cells. A clathrin inhibitor (pitstop 2) impeded the early replication of IBDV, even when ARF6 was overexpressed. These results indicated that ARF6 promotes viral replication by affecting the internalization of IBDV, which may involve clathrin-dependent endocytosis. Our findings improve the understanding of the processes governing IBDV infection and provide insights into its prevention and control.

Systematic assessment of the antifungal mechanism of soil fumigant methyl isothiocyanate against Fusarium oxysporum.

Fusarium oxysporum is an important phytopathogenic fungus, it can be controlled by the soil fumigant methyl isothiocyanate (MITC). However, the antimicrobial mechanism of MITC against F. oxysporum, especially at the transcriptional level, is still unclear. In this experiment, the antimicrobial mechanism of MITC against F. oxysporum was investigated. Our results indicated that when F. oxysporum was exposed to 6 mg/L MITC for 12 h, the inhibitory rate of MITC on F. oxysporum was 80%. Transmission electron microscopes showed that the cell wall and membrane of F. oxysporum had shrunk and folded, vacuoles increased, and mitochondria swelled and deformed. In addition, the enzyme activity of F. oxysporum treated with MITC showed a decrease of 32.50%, 8.28% and 74.04% in catalase, peroxidase and superoxide dismutase, respectively. Transcriptome sequencing of F. oxysporum was performed and the results showed that 1478 differentially expressed genes (DEGs) were produced in response to MITC exposure. GO and KEGG analysis showed that the DEGs identified were involved in substance and energy metabolism, signal transduction, transport and catalysis. MITC disrupted cell homeostasis by influencing the expression of some key genes involved in chitin synthase and detoxification enzymes production, but F. oxysporum also protected itself by up-regulating genes involved in energy synthesis (such as upregulating acnA, CS and LSC2 in TCA). qRT-PCR data validated the reliability of transcriptome data. Our research used biochemical and genetic techniques to identify molecular lesions in the mycelia of F. oxysporum exposed to MITC, and provide valuable insights into the toxic mechanism of pathogenic fungi mediated by MITC. These techniques are also likely to be useful for rapidly screening and identifying new, environmentally-friendly soil fumigants that are efficacious against fungal pathogens.

Strain and Shell Thickness Engineering in Pd3 Pb@Pt Bifunctional Electrocatalyst for Ethanol Upgrading Coupled with Hydrogen Production.

The ethanol oxidation reaction (EOR) is an attractive alternative to the sluggish oxygen evolution reaction in electrochemical hydrogen evolution cells. However, the development of high-performance bifunctional electrocatalysts for both EOR and hydrogen evolution reaction (HER) is a major challenge. Herein, the synthesis of Pd3 Pb@Pt core-shell nanocubes with controlled shell thickness by Pt-seeded epitaxial growth on intermetallic Pd3 Pb cores is reported. The lattice mismatch between the Pd3 Pb core and the Pt shell leads to the expansion of the Pt lattice. The synergistic effects between the tensile strain and the core-shell structures result in excellent electrocatalytic performance of Pd3 Pb@Pt catalysts for both EOR and HER. In particular, Pd3 Pb@Pt with three Pt atomic layers shows a mass activity of 8.60 A mg-1 Pd+Pt for ethanol upgrading to acetic acid and close to 100% of Faradic efficiency for HER. An EOR/HER electrolysis system is assembled using Pd3 Pb@Pt for both the anode and cathode, and it is shown that low cell voltage of 0.75 V is required to reach a current density of 10 mA cm-2 . The present work offers a promising strategy for the development of bifunctional catalysts for hybrid electrocatalytic reactions and beyond.

Integrative multiomics profiling of passion fruit reveals the genetic basis for fruit color and aroma.

Passion fruit (Passiflora edulis) possesses a complex aroma and is widely grown in tropical and subtropical areas. Here, we conducted the de novo assembly, annotation, and comparison of PPF (P. edulis Sims) and YPF (P. edulis f. flavicarpa) reference genomes using PacBio, Illumina, and Hi-C technologies. Notably, we discovered evidence of recent whole-genome duplication events in P. edulis genomes. Comparative analysis revealed 7.6∼8.1 million single nucleotide polymorphisms, 1 million insertions/deletions, and over 142 Mb presence/absence variations among different P. edulis genomes. During the ripening of yellow passion fruit, metabolites related to flavor, aroma, and color were substantially accumulated or changed. Through joint analysis of genomic variations, differentially expressed genes, and accumulated metabolites, we explored candidate genes associated with flavor, aroma, and color distinctions. Flavonoid biosynthesis pathways, anthocyanin biosynthesis pathways, and related metabolites are pivotal factors affecting the coloration of passion fruit, and terpenoid metabolites accumulated more in PPF. Finally, by heterologous expression in yeast (Saccharomyces cerevisiae), we functionally characterized 12 terpene synthases. Our findings revealed that certain TPS homologs in both YPF and PPF varieties produce identical terpene products, while others yield distinct compounds or even lose their functionality. These discoveries revealed the genetic and metabolic basis of unique characteristics in aroma and flavor between the 2 passion fruit varieties. This study provides resources for better understanding the genome architecture and accelerating genetic improvement of passion fruits.

Non-contrast-enhanced MR angiography of left gastric vein in patients with gastroesophageal varices: morphology and blood supply analysis.

OBJECTIVES: To investigate the feasibility of non-contrast-enhanced MR angiography (NCE-MRA) in evaluating the morphology and blood supply of left gastric vein (LGV) in patients with gastroesophageal varices. METHODS: Between March 2021 and October 2022, patients with gastroesophageal varices and who underwent NCE-MRA were retrospectively reviewed. In order to evaluate the blood supply of LGV, superior mesenteric vein (SMV) and splenic vein (SV) were visualized separately by using inflow-sensitive inversion recovery sequence. Two radiologists independently assessed the image quality, determined the origination and the blood supply of LGV, and measured the diameter of LGV. The origination and diameter of LGV were compared between NCE-MRA and contrast-enhanced CT. Differences in blood supply were compared between LGVs with different originations. RESULTS: A total of 53 patients were enrolled in this study and the image quality was categorized as good or excellent in 52 patients. No significant differences were observed in visualizing the origination and the diameter of LGV between NCE-MRA and contrast-enhanced CT (p > .05). The blood supply of LGV was related to its origination (p < .001). Most LGVs with SV origination were supplied by SV. If LGV was originated from the portal vein (PV), about 70% of them were supplied by both SV and SMV. Compared with LGVs with SV origination, LGVs with PV origination showed more chance to receive blood from SMV (p < .001). CONCLUSION: Non-contrast-enhanced MR angiography appears to be a reliable technique in evaluating the morphology and blood supply of LGV in patients with gastroesophageal varices. CLINICAL RELEVANCE STATEMENT: Non-contrast-enhanced MR angiography provides valuable information for the management of gastroesophageal varices. Especially, it benefits patients with renal insufficiency. KEY POINTS: • Non-contrast-enhanced MR angiography using inflow-sensitive inversion recovery technique can be used for evaluating not only morphology as CT but also blood supply of left gastric vein. • The blood supply of left gastric vein is related to its origination and left gastric vein with portal vein origination shows more chance to receive blood from superior mesenteric vein.

Anthracnose of Macleaya cordata Caused by Colletotrichum aenigma in China.

Macleaya cordata (Willd.) R. Br. is a perennial herbaceous medicinal plant (Papaveraceae) commercially cultivated in China which has been studied for detumescence, detoxification, and insecticidal effect (Lin et al. 2018). In August 2021, anthracnose was observed in 2-year-old M. cordata plants in Benxi county, northeast China (41°45'48″N, 123°69'15″E). Dozens of irregular reddish-brown spots (3-11 mm) were observed on each diseased leaf. The lesions were covered with a layer of gray-white mycelia. As the disease progressed, the spots became necrosis and perforation or they would merged into large lesions, ultimately resulting in wilted leaves (Fig. 1). More than 33% of the plants in a 16-ha field were infected in 2021. The diseased leaves were collected and cut into 3-8 mm pieces, surface-disinfested by immersing them into 1% NaOCl for 2 min, and rinsed three times with sterile distilled water. They were then dried with sterilized absorbent paper, placed on PDA medium amended with chloramphenicol (40 mg/L), and incubated in darkness at 25°C with a 12-h photoperiod. Twenty isolates (BLH1 to 20) were obtained and purified using a single-spore method. Isolate BLH12 was identified and used for the pathogenicity test. Colonies were sparsely fluffy with smooth edges, and gradually became gray to pale orange from the initial white. The underside of the colonies was pale orange towards the center. Conidia were single-celled, cylindrical, and transparent with broadly blunt ends, measuring (15.13 ± 1.14) × (5.80 ± 0.60) µm (n=50). Appressoria were single-celled, brown-to-dark brown, usually elliptical or irregular, and sometimes lobed. Setae were not observed. The isolate was initially identified as Colletotrichum gloeosporioides complex (Prihastuti et al. 2009). The identification was confirmed as described previously (Weir et al. 2012). The rDNA internal transcribed spacer region (OP415560), the glyceraldehyde-3-phosphate dehydrogenase (OP433642), chitin synthase (OP433643), calmodulin (OP433644), actin (OP433645), glutamine synthetase (OP433646), ß-tubulin (OP433647), and superoxide dismutase (OP433648) gene sequences were obtained (Carbone & Kohn 1999; Weir et al. 2012), and BLAST searches revealed 99-100% homology with the type culture ICMP 18608 (JX010244, JX010044, JX009683, JX009443, JX009744, JX010078, JX010389, and JX010311). A phylogenetic analysis of combining all loci indicated BLH12 and the type strain of C. aenigma were clustered in one group (Fig. 2). Based on the basis of morphological characteristics and phylogenetic relationships, BLH12 was identified as C. aenigma. For the pathogenicity test, healthy 2-year-old plants were sprayed with a BLH12 spore suspension (1 × 105/mL). Control plants were sprayed with sterile water.There were three replicates (five plants each) per treatment. All plants were incubated at 25°C (12-h photoperiod and 86% relative humidity) and examined after 7 days. The experiment was repeated twice. The inoculated plants showed lesions on the leaf surface, similar to those in the field, whereas the control plants were asymptomatic. The pathogen was successfully reisolated and identified as the methods mentioned above. This fungus reportedly infects the leaves of many woody plants in China (Wang et al. 2020; Zhang et al. 2021). This is the first report of C. aenigma causing anthracnose on M. cordata, which will provide an guideline for developing effective field control practices for the disease.

First Report of Calonectria montana Causing Leaf Spot on Ligularia fischeri in China.

Ligularia fischeri (Ledeb.) is a perennial herbal plant of Compositae that is cultivated commercially in China as a medicinal, ornamental, and edible plant. Leaf spots were observed in 2-year-old L. fischeri in Benxi County of northeast China, in August 2021. Irregular reddish brown spots ranging from 3 to 11 mm were observed on infected leaves, and each leaf had dozens of spots (Fig. 1). As the disease progressed, the diseased spots withered and the centers fell out, and multiple lesions merge into large diseased spots, causing leaf wilting. The roots and stem bases were not infected during the reproductive stage. More than 37% of the plants in a 18 ha field were infected in 2021. The ten diseased leaves were collected and cut into small (3-5 mm) pieces, which were surface-disinfested by immersing into 1% NaOCl for 2 min and rinsing with sterile distilled water three times. The leaf pieces were then placed on acidified potato dextrose agar (PDA) in petri plates and incubated in the dark at 25°C. Twenty isolates with the same morphological characteristics were obtained. Isolates were further purified by starting a new colony for each isolate from a single spore collected from water agar. Isolate TYTW7 was randomly selected for identification and pathogenicity testing. It grew rapidly and produced profuse aerial mycelia with a carmine red underside. The conidiophores had many fertile branches and formed an elongated stipe with a sphaeropedunculate vesicle at the tip. The one-septate conidia were cylindrical and almost straight with parallel walls and rounded ends. Their sizes ranged from 30.35 to 51.76 × 2.93 to 5.01 µm (n = 100) and the pathogens were initially identified as Calonectria sp. (Crous 2002; Crous et al. 2004; Lombard et al. 2015, 2016). Further confirmation of the identification was determined according to published method (Liu and Chen 2017; Shao and Li 2021). The partial gene regions including the translation elongation factor 1-alpha (GenBank accession no. OP290551), histone H3 (OP290552), calmodulin (OP290553) and ß-tubulin (OP290554) were obtained, and BLAST searches showed 99-100% homology with the ex-type culture CERC 8952 (MF527049, MF527065, MF527081 and MF527107) and phylogenetic analysis combining all loci revealed that the isolate TYTW7 and the type strain of Ca. montana clustered in one group (Fig. 2). Based on morphological characteristics and phylogenetic analysis, isolate TYTW7 was identified as Ca. Montana. Healthy 2-year-old plants were used for the pathogenicity test. A spore suspension (1×105 spores/mL water) was used to inoculate three host plants; sterile water was sprayed on the same number plants serving as a control. The experiment was repeated three times. All plants were incubated at 27±2°C (12h photoperiod) and were evaluated after seven days. The inoculated plants showed lesions on the leaf surface, similar to those in the field, and the control remained symptomless. The pathogens were successfully reisolated and identified by sequencing, and no pathogens were isolated from symptomless control plants. To our knowledge, this is the first report of Ca. montana causing L. fischeri leaf spot. The disease poses a threat to the production and more control strategies are needed on management options to minimize losses.

Anthracnose of Brachybotrys paridiformis Caused by Colletotrichum siamense in Northeast China.

Brachybotrys paridiformis Maxim. ex Oliv. (Boraginaceae) is a perennial medicinal plant and vegetable that is cultivated commercially in China. Anthracnose is a devastating disease of B. paridiformis, with annual production losses exceeding 33% based on our survey. In July 2021, anthracnose of B. paridiformis was observed on 2-year-old plants in Shenyang city, Northeast China, which is the most important region for B. paridiformis cultivation. Round or irregular-shaped black spots were exhibited on leaves, with the leaf edges most commonly infected. As the necrosis expanded, the leaves withered and dropped; young leaves were generally not infected (Fig. 1). More than 40% of the plants in a 21-ha sampling field were infected in 2021. Symptomatic leaves (n = 20) were collected and the diseased tissue was cut into small pieces, immersed in 1% NaOCl for 2 min, rinsed three times with sterile water, and placed on acidified potato dextrose agar (PDA) in Petri dishes. After a 3-day incubation in darkness at 25 °C, 18 suspected single-pure morphologically identical Colletotrichum isolates were obtained and sequenced. Isolate SQZ9 was randomly selected and identified. Colonies on PDA were initially white, but gradually became pale brownish with a reverse side that was pale yellowish to pinkish. Aerial mycelia were grayish-white, dense, and cottony, with microsclerotia detected on some aging mycelia. The detected single-celled conidia (11.65-17.25 × 4.25-6.15 µm; n = 50) were fusiform to cylindrical with obtuse to slightly rounded ends. Appressoria were ovoid to clavate and medium brown. Setae were not observed. The morphological characteristics were similar to those of Colletotrichum spp. (Prihastuti et al. 2009; Weir et al. 2012). Initial BLAST searches of the GenBank database revealed the SQZ9 rDNA internal transcribed spacer region (OP389109, 566 bp), glyceraldehyde-3-phosphate dehydrogenase (OP407730, 260 bp), chitin synthase (OP407731, 301 bp), calmodulin (OP407732, 712 bp), actin (OP407733, 282 bp), glutamine synthetase (OP407734, 909 bp), ß-tublin (OP407735, 498 bp), and superoxide dismutase (OP407736, 396 bp) sequences were respectively 99%-100% similar to the C. siamense type strain JX010278, JX010019, JX009709, GQ856775, GQ856730, JX010100, JX010410, and JX010332 sequences (Carbone & Kohn 1999; Moriwaki & Tsukiboshi 2009; Stephenson et al. 1997). The SQZ9 identity was confirmed by constructing a phylogenetic tree combining all loci, which grouped the isolate and the C. siamense type strain in the same clade (Fig. 2). For pathogenicity tests, 15 healthy 2-year-old plants (3 plants per pot) were spray-inoculated with SQZ9 conidial suspension (1 × 105 conidia/mL) at 2 mL per plant. Same number of plants sprayed with water were used as control. This experiment was repeated twice. All plants were covered with clear plastic bags for 72 h to maintain high humidity and then placed in a greenhouse (29 °C, natural light, and 85% relative humidity). After six days, the inoculated leaves exhibited symptoms that were similar to those observed in the field, but the controls were symptomless. The same fungus was recovered from inoculated symptomatic leaves, and its identity was confirmed by sequencing and a phylogenetic analysis. This is the first report of C. siamense causing anthracnose on B. paridiformis in China. Future studies should assess the effectiveness of chemical and biological control measures for managing this disease.

Noninvasive Prediction of Sperm Retrieval Using Diffusion Tensor Imaging in Patients with Nonobstructive Azoospermia.

Microdissection testicular sperm extraction (mTESE) is the first-line treatment plan for nonobstructive azoospermia (NOA). However, studies reported that the overall sperm retrieval rate (SRR) was 43% to 63% among men with NOA, implying that nearly half of the patients fail sperm retrieval. This study aimed to evaluate the diagnostic performance of parameters derived from diffusion tensor imaging (DTI) in predicting SRR in patients with NOA. Seventy patients diagnosed with NOA were enrolled and classified into two groups based on the outcome of sperm retrieval during mTESE: success (29 patients) and failure (41 patients). Scrotal magnetic resonance imaging was performed, and the DTI parameters, including mean diffusivity and fractional anisotropy, were analyzed between groups. The results showed that there was a significant difference in mean diffusivity values between the two groups, and the area under the curve for mean diffusivity was calculated as 0.865, with a sensitivity of 72.2% and a specificity of 97.5%. No statistically significant difference was observed in fractional anisotropy values and sex hormone levels between the two groups. This study demonstrated that the mean diffusivity value might serve as a useful noninvasive imaging marker for predicting the SRR of NOA patients undergoing mTESE.