PTOV1-AS1 desensitizes colorectal cancer cells to 5-FU through depressing miR-149-5p to activate the positive feedback loop with Wnt/ß-catenin pathway.

5-Fluorouracil is a commonly used chemotherapy drug for colorectal cancer. Resistance to 5-Fluorouracil remains a challenge. This research aimed to explore the mechanism of 5-Fluorouracil resistance in colorectal cancer. RT-qPCR and Western blot were used to determine the RNA and protein expression in both cells and exosome. Assays in vitro and in vivo were performed to measure the role of miR-149-5p in colorectal cancer cells. RIP, luciferase activity report, and RNA pulldown assay were applied to detect the association of PTOV1-AS1, SUV39H1, miR-149-5p, and FOXM1. MiR-149-5p was down-expressed in 5-Fluorouracil-resistant cells. MiR-149-5p enhanced the effectiveness of 5-Fluorouracil both in vitro and in vivo. Sensitive colorectal cancer cells released exosomal miR-149-5p to sensitize resistant cells to chemotherapy. Mechanistically, miR-149-5p targeted the FOXM1 to inactivate Wnt/ß-catenin pathway, and PTOV1-AS1 recruited SUV39H1 to suppress miR-149-5p transcription, in turn activating Wnt/ß-catenin pathway, and forming a positive feedback loop with FOXM1. PTOV1-AS1 inhibits miR-149-5p by a positive feedback loop with FOXM1-mediated Wnt/ß-catenin pathway, which provides insights into a potential novel target for enhancing the effectiveness of chemotherapy in colorectal cancer patients.

RPA-CRISPR/Cas12a-mediated isothermal amplification for rapid detection of Phytopythium helicoides.

Phytopythium helicoides, which belongs to the algae (Chromista), Oomycota, Pythiales, Pythiaceae and Phytophthora, is a quarantine pathogen that causes brown rot of fruits, stem rot and root rot, along with other symptoms that can damage several tree species in urban landscaping. Therefore, disease management requires rapid and accurate diagnosis. The present study used recombinase polymerase amplification (RPA) in conjunction with the CRISPR/Cas12a system to identify P. helicoides. The test exhibited high specificity and sensitivity and could detect 10 pg.µL-1 of P. helicoides genomic DNA at 37 ℃ within 20 minutes. The test results were visible by excitation of fluorophores by blue light. This groundbreaking test is able to detect P. helicoides in artificially inoculated Rhododendron leaves. The RPA-CRISPR/Cas12a detection assay developed in this study is characterized by its sensitivity, efficiency, and convenience. Early detection and control of P. helicoides is crucial for the protection of urban green cover species.

A rapid, equipment-free method for detecting avirulence genes of Pyricularia oryzae using a lateral flow strip-based RPA assay.

Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. Using resistant rice varieties is the most cost-effective way to control rice blast. Consequently, it is critical to monitor the distribution frequency of avirulence genes in rice planting field to facilitate the breedings of resistant rice varieties. In this study, we established a rapid RPA-LFD detection system for the identification of AvrPik, Avr-Piz-t and Avr-Pi9. The optimized reaction temperature and duration were 37°C and 20 min, indicating that the reaction system could be initiated by body temperature without relying on any precision instruments. Specificity analysis showed that the primer and probe combinations targeting three Avr genes exhibited a remarkable specificity for at genus-level detection. Under the optimized condition, the lower detected thresholds of AvrPik, Avr-Piz-t and Avr-Pi9 were 10 fg/µl, 100 fg/µl and 10 pg/µl, respectively. Notably, the detection sensitivity of three Avr genes was much higher than that of PCR. In addition, we also successfully detected the presence of AvrPik, Avr-Piz-t and Avr-Pi9 in the leaf and panicle blast lesions with the RPA-LFD detection system. In particular, the genomic DNA was extracted using the simpler PEG-NaOH rapid extraction method. In summary, we developed the RPA detection system for AvrPik, Avr-Pi9 and Avr-Piz-t, combined with the PEG-NaOH rapid DNA extraction method. The innovative approach achieved rapid, real-time and accurate detection of three Avr genes in the field, which is helpful to understand the distribution frequency of the three Avr genes in the field and provide theoretical reference for the scientific layout of rice resistant varieties.

Stromal Vascular Fraction Gel (SVF-Gel) Combined with Nanofat for Tear Trough Deformity.

BACKGROUND: Tear trough deformity makes patients appear tired. Patients with less severe tear trough deformity prefer a less invasive method to correct the deformity. The infraorbital area is a multilayered tissue, and the aging of various components leads to tear trough deformity. To this end, we utilized the different characteristics of different fat derivatives to correct tear trough deformity. METHODS: Thirty-two patients with Barton Grade I/II tear trough deformity were enrolled in this study between September 2020 and March 2021. We injected Stromal Vascular Fraction Gel (SVF-Gel) into the suborbicularis oculi fat layer and Nanofat into the subcutaneous. After 12 months of follow-up, we evaluated the changes using standardized clinical photogrammetric techniques, volume, global aesthetic improvement scale, and patient self-evaluation. RESULTS: There were no major complications in any of the 32 patients. The measured data points demonstrated improvements in all aesthetic parameters. The width of the tear trough and the distance from the pupil to the tear trough improved. The Global Aesthetic Improvement Scale (GAIS) showed a high score (2.45±0.64 points), with patient self-assessment showing satisfactory results. CONCLUSION: SVF-Gel combined with Nanofat injection can effectively correct tear trough deformities. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Development and Validation of a Patient Discharge Readiness Scale for Daytime Cataract Surgery (DRS-CAT).

PURPOSE: To ensure the safety of patients discharged from the hospital, a nurse-assessed scale for outpatient cataract surgery patients was constructed to provide a special tool for cataract patients' discharge readiness evaluation. DESIGN: This is a methodological study. METHODS: The development of the tool was completed between 2021 and 2022. Based on the literature review and qualitative interviews, the initial entry pool of the discharge readiness scale was established. After consultation with Delphi experts, the preliminary scale was tested by 312 participants to screen items and test reliability and validity. The analysis included internal consistency, content validity, and construct validity. The Strengthening the Reporting of Observation studies in Epidemiology (STROBE) checklist was used as the reporting guideline for this study. FINDINGS: The final Discharge Readiness Scale for Cataract surgery consists of 21 items in five dimensions: cognition of discharge readiness, personal status, mastery of health education knowledge, coping capacity, and social support. Five common factors were extracted from the exploratory factor analysis, and they explained 70.12% of the total variance. All of the indexes of the confirmatory factor analysis were within the theoretical allowable range. The Cronbach's α of the total scale was 0.903, and the scale-level content validity index/average variance extracted was 0.99. CONCLUSIONS: The Discharge Readiness Scale for Cataract surgery, evaluated by nurses, has good reliability and validity and can be used to determine the discharge readiness of cataract patients undergoing day surgery.

Dithienoquinoxalineimide-Based Polymer Donor Enables All-Polymer Solar Cells Over 19 % Efficiency.

All-polymer solar cells (all-PSCs) have been regarded as one of the most promising candidates for commercial applications owing to their outstanding advantages such as mechanical flexibility, light weight and stable film morphology. However, compared to large amount of new-emerging excellent polymer acceptors, the development of high-performance polymer donor lags behind. Herein, a new D-π-A type polymer donor, namely QQ1, was developed based on dithienoquinoxalineimide (DTQI) as the A unit, benzodithiophene with thiophene-conjugated side chains (BDTT) as the D unit, and alkyl-thiophene as the π-bridge, respectively. QQ1 not only possesses a strong dipole moment, but also shows a wide band gap of 1.80 eV and a deep HOMO energy level of -5.47 eV, even without halogen substituents that are commonly indispensable for high-performance polymer donors. When blended with a classic polymer acceptor PY-IT, the QQ1-based all-PSC delivers an outstanding PCE of 18.81 %. After the introduction of F-BTA3 as the third component, a record PCE of 19.20 % was obtained, the highest value reported so far for all-PSCs. The impressive photovoltaic performance originates from broad absorption range, reduced energy loss, and compact π-π stacking. These results provide new insight in the rational design of novel nonhalogenated polymer donors for further development of all-PSCs.

Optimizing Molecular Crystallinity and Suppressing Electron-Phonon Coupling in Completely Non-Fused Ring Electron Acceptors for Organic Solar Cells.

High open-circuit voltage (Voc) organic solar cells (OSCs) have received increasing attention because of their promising application in tandem devices and indoor photovoltaics. However, the lack of a precise correlation between molecular structure and stacking behaviors of wide band gap electron acceptors has greatly limited its development. Here, we adopted an asymmetric halogenation strategy (AHS) and synthesized two completely non-fused ring electron acceptors (NFREAs), HF-BTA33 and HCl-BTA33. The results show that AHS significantly enhances the molecular dipoles and suppresses electron-phonon coupling, resulting in enhanced intramolecular/intermolecular interactions and decreased nonradiative decay. As a result, PTQ10 : HF-BTA33 realizes a power conversion efficiency (PCE) of 11.42 % with a Voc of 1.232 V, higher than that of symmetric analogue F-BTA33 (PCE=10.02 %, Voc=1.197 V). Notably, PTQ10 : HCl-BTA33 achieves the highest PCE of 12.54 % with a Voc of 1.201 V due to the long-range ordered π-π packing and enhanced surface electrostatic interactions thereby facilitating exciton dissociation and charge transport. This work not only proves that asymmetric halogenation of completely NFREAs is a simple and effective strategy for achieving both high PCE and Voc, but also provides deeper insights for the precise molecular design of low cost completely NFREAs.

Bioluminogenic Probe for Rapid, Ultrasensitive Detection of ß-Lactam-Resistant Bacteria.

Increasingly difficult-to-treat infections by antibiotic-resistant bacteria have become a major public health challenge. Rapid detection of common resistance mechanisms before empiric antibiotic usage is essential for optimizing therapeutic outcomes and containing further spread of resistance to antibiotics among other bacteria. Herein, we present a bioluminogenic probe, D-Bluco, for rapid detection of ß-lactamase activity in viable pathogenic bacteria. D-Bluco is a pro-luciferin caged by a ß-lactamase-responsive cephalosporin structure and further conjugated with a dabcyl quencher. The caging and quenching significantly decreased the initial background emission and increased the signal-to-background ratio by more than 1200-fold. D-Bluco was shown to detect a broad range of ß-lactamases at the femtomolar level. An ultrasensitive RAPID bioluminescence assay using D-Bluco can detect 102 to 103 colony forming unit per milliliter (cfu/mL) of ß-lactamase-producing Enterobacterales in urine samples within 30 min. The high sensitivity and rapid detection make the assay attractive for the use of point-of-care diagnostics for lactam-resistant pathogens.

Asymmetric supercapacitors based on SnNiCoS ternary metal sulfide electrodes.

While metal sulfides have extensively investigated as electrode materials for supercapacitors, the further optimization of their material system is still necessary to achieve satisfied performance. In this work, we reported the synthesis of ternary metal sulfide SnNiCoS and its application as electrode material of asymmetric supercapacitors, in which active carbon is used as the other electrode. For control experiments, asymmetric supercapacitors based on single metal sulfide CoS and binary metal sulfide NiCoS are also fabricated and investigated. The results show that the nanospherical SnNiCoS achieves the best performance. Ternary sulphide materials offer more redox than corresponding single-metal sulphides due to the synergy among various transition metal elements. The specific capacitance is 18.6 F cm-2at current density of 5 mA·cm-2. An energy density of 937.2µWh cm-2is achieved at a power density of 4000µW·cm-2. After 8000 cycles, the capacity retention rate is 82.9%. Present work indicates that SnNiCoS ternary metal sulfide could be a promising composite for high performance supercapacitors.

Exposure to Fine Particulate Matter Constituents and Human Semen Quality Decline: A Multicenter Study.

Exposure to fine particulate matter (PM < 2.5 µm in diameter [PM2.5]) may accelerate human sperm quality decline, although research on this association is limited. Our objective was to investigate the relationship between exposure to the chemical constituents of PM2.5 air pollution and decreased sperm quality and to further explore the exposure-response relationship. We conducted a multicenter population-based cohort study including 78,952 semen samples from 33,234 donors at 6 provincial human sperm banks (covering central, northern, southern, eastern, and southwestern parts of China) between 2014 and 2020. Daily exposure to PM2.5 chemical composition was estimated using a deep learning model integrating a density ground-based measure network at a 1 km resolution. Linear mixed models with subject- and center-specific intercepts were used to quantify the harmful impacts of PM2.5 constituents on semen quality and explore their exposure-response relationships. Per interquartile range (IQR) increase in PM2.5 exposure levels during spermatogenesis was significantly associated with decreased sperm concentration, progressive motility, and total motility. For PM2.5 constituents, per IQR increment in Cl- (ß: -0.02, 95% CI: [-0.03, -0.00]) and NO3- (ß: -0.05, 95% CI: [-0.08, -0.02]) exposure was negatively associated with sperm count, while NH4+ (ß: -0.03, 95% CI: [-0.06, -0.00]) was significantly linked to decreased progressive motility. These results suggest that exposure to PM2.5 chemical constituents may adversely affect human sperm quality, highlighting the urgent need to reduce PM2.5 exposure.

A Frontline, Rapid, Nucleic Acid-Based Fusarium circinatum Detection System Using CRISPR/Cas12a Combined with Recombinase Polymerase Amplification.

Pitch canker caused by the fungus Fusarium circinatum is a damaging disease that affects pines in Europe, South Africa, and North America in both the southeast and west coast of the United States. Several countries, including China, have listed F. circinatum as a quarantine pathogen. Timely detection, an important pillar of the quarantine effort, can efficiently prevent the introduction of F. circinatum into new areas or facilitate management and eradication strategies in already infested sites. In this study, we developed an F. circinatum detection technique based on a combination of recombinase polymerase amplification (RPA) with CRISPR/Cas12a technology (termed RPA-CRISPR/Cas12a). After obtaining DNA, this novel method can be utilized for the molecular identification of F. circinatum using the naked eye and can specifically detect F. circinatum at DNA concentrations as low as 200 fg within 30 min at 37°C. The system is sensitive for both standard laboratory samples and samples from the field. In summary, we have developed a simple, rapid, sensitive, unaided-eye visualization, RPA-CRISPR/Cas12a-based detection system for the molecular identification of F. circinatum that does not require technical expertise or expensive ancillary equipment.

Recombinase Polymerase Amplification-Lateral Flow Dipstick Assay for Rapid Detection of Fusarium circinatum Based on a Newly Identified Unique Target Gene.

Pitch canker caused by the fungus Fusarium circinatum is an important disease affecting pine trees in Europe and South Africa. Several countries, including China, have listed F. circinatum as a quarantine pathogen. Therefore, timely detection of F. circinatum could efficiently prevent its introduction into new areas or facilitate spread management in already infected sites. In this study, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay was developed for rapid detection of F. circinatum based on a new target gene, Fcir2067, identified from whole-genome sequences. The assay was highly specific to F. circinatum. In fact, it exclusively detected F. circinatum isolates; 53 isolates of fungal and oomycete species and 2 nematodes of Bursaphelenchus xylophilus and B. mucronatus were not detected. By detecting as little as 10 pg of F. circinatum genomic DNA in a 50-µl reaction, the RPA-LFD assay was 10 times more sensitive than conventional PCR assays. F. circinatum was also detected in artificially inoculated pine needles of Cedrus deodara. These results demonstrated that the developed RPA-LFD assay has the potential for rapid detection of F. circinatum in regions at high risk of infection. The RPA-LFD assay might serve as an alternative method for the early detection of F. circinatum.

Experimental Study on the Biological Outcome of Auricular Cartilage and Costal Cartilage at Different Time Periods After Autologous Cartilage Rhinoplasty.

Since autologous cartilage is a good transplant material, it is widely used in various fields of clinical medicine. In this study, we collected clinical specimens obtained at different numbers of years after transplantation and used histologic staining to explore the post-transplantation changes in auricular cartilage and costal cartilage. A retrospective analysis was performed on patients who underwent primary autologous cartilage rhinoplasty and secondary rhinoplasty from 2017 to 2021, and the remaining autologous cartilage tissue after surgery was used for histologic testing. As time progressed after transplantation, the density of costal chondrocytes decreased first and then increased, while the secretion of type II collagen and extracellular matrix both decreased slightly. There was a clear boundary between the cartilage tissue and the surrounding connective tissue, and there was no ingrowth of blood vessels in the cartilage. Auricular cartilage showed a decrease in the integrity of the matrix edge. Moreover, local fibrosis was visible, and vascular ingrowth was observed at the edge of the cartilage. The content of type II collagen first increased and then decreased, and the cell secretion function was lower than that of normal chondrocytes. The results of the study suggest that the histologic outcome of elastic cartilage after transplantation is significantly different from that of hyaline cartilage. Moreover, costal cartilage was more stable than auricular cartilage after transplantation.

Biological Changes of Autologous Auricular Cartilage in Rhinoplasty.

Autologous auricular cartilage is used extensively as a good graft material in rhinoplasty. In this study, clinical specimens from patients who underwent revision rhinoplasty with auricular cartilage grafts were collected to compare the changes before and after auricular cartilage transplantation with the use of histologic, immunohistochemical, and quantitative assays. Patients who underwent revision rhinoplasty from 2018 to 2022 were analyzed. Fresh auricular cartilage left after surgery and auricular cartilage graft tissue were examined and compared. Compared with fresh auricular cartilage, local fibrosis was seen in the transplanted auricular cartilage with a slight decrease in elastic fibers, type II collagen, and extracellular matrix secretion. Quantitative assays showed a decrease in glycosaminoglycan, DNA, and total collagen content in the transplanted auricular cartilage tissue. The results of the study suggest that the histologic characteristics, cell functionality, and biochemical composition of the grafted cartilage changed to a certain extent after autologous auricular cartilage graft rhinoplasty. These results provide insights into the selection of graft/filler materials for rhinoplasty and what changes to expect.

The Spectrum of Low-pTJ/ψ in Heavy-Ion Collisions in a Statistical Two-Body Fractal Model.

We establish a statistical two-body fractal (STF) model to study the spectrum of J/ψ. J/ψ serves as a reliable probe in heavy-ion collisions. The distribution of J/ψ in hadron gas is influenced by flow, quantum and strong interaction effects. Previous models have predominantly focused on one or two of these effects while neglecting the others, resulting in the inclusion of unconsidered effects in the fitted parameters. Here, we study the issue from a new point of view by analyzing the fact that all three effects induce a self-similarity structure, involving a J/ψ-π two-meson state and a J/ψ, π two-quark state, respectively. We introduce modification factor qTBS and q2 into the probability and entropy of charmonium. qTBS denotes the modification of self-similarity on J/ψ, q2 denotes that of self-similarity and strong interaction between c and c¯ on quarks. By solving the probability and entropy equations, we derive the values of qTBS and q2 at various collision energies and centralities. Substituting the value of qTBS into distribution function, we successfully obtain the transverse momentum spectrum of low-pT J/ψ, which demonstrates good agreement with experimental data. The STF model can be employed to investigate other mesons and resonance states.

Metabolite profiling analysis of plasma, urine, and feces of rats after oral administration of Flos Chrysanthemi Indici preparation through UHPLC-Q-Exactive-MS combined with pharmacokinetic study of markers by UHPLC-QQQ-MS/MS.

Zhenju Antihypertensive Tablet is one of the Flos Chrysanthemi Indici preparations (FCIP) with antihypertensive effect. In order to investigate its process in vivo, a method using ultra-high performance liquid chromatography coupled with quadruple Exactive mass spectrometry (UHPLC-Q-Exactive-MS) was established for comprehensive analysis of the absorbed active component and metabolites in rat plasma, urine, and feces after the oral administration of FCIP. As a result, a total of 61 FCIP-related xenobiotics were identified, including 35 in plasma (25 prototypes, 10 metabolites), 40 in urine (23 prototypes, 17 metabolites), and 25 in feces (12 prototypes, 13 metabolites). The metabolism reactions included phase I reactions (such as oxidation, methylation, and reduction) and phase II reactions (such as sulfate conjugation and glucuronide conjugation). Meanwhile, a verified and optimized ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) method was developed for the simultaneous investigation of the pharmacokinetic profiles of four markers in FCIP. The results showed that all of the four components had good oral absorption effect. This study simultaneously investigated the comprehensive metabolic profiling of FCIP in rat plasma, urine, and feces after the oral administration as well as the four components pharmacokinetic behavior. The results can provide the therapeutic material basis for FCIP.

The Subtle Structure Modulation of A2 -A1 -D-A1 -A2 Type Nonfullerene Acceptors Extends the Photoelectric Response for High-Voltage Organic Photovoltaic Cells.

Molecular structural modifications are utilized to improve the short-circuit current (JSC ) of high-voltage organic photovoltaics (OPVs). Herein, the classic non-fullerene acceptor (NFA), BTA3, is chosen as a benchmark, with BTA3b containing the linear alkyl chains on the middle core and JC14 fusing thiophene on the benzotriazole (BTA) unit as a contrast. The photovoltaic devices based on J52-F: BTA3b and J52-F: JC14 achieve wider external quantum efficiency responses with band edges of 730 and 800 nm, respectively than that of the device based on J52-F: BTA3 (715 nm). The corresponding  JSC increases to 14.08 and 15.78 mA cm-2 , respectively, compared to BTA3 (11.56 mA cm-2 ). The smaller Urbach energy and higher electroluminescence efficiency guarantee J52-F: JC14 a decreased energy loss (0.528 eV) and a high open-circuit voltage (VOC ) of 1.07 V. Finally, J52-F: JC14 combination achieves an increased power conversion efficiency (PCE) of 10.33% than that of J52-F: BTA3b (PCE = 9.81%) and J52-F: BTA3 (PCE = 9.04%). Overall, the research results indicate that subtle structure modification of NFAs, especially introducing fused rings, is a simple and effective strategy to extend the photoelectric response, boosting the  JSC and ensuring a high VOC beyond 1.0 V.

First Report of Crown and Root Rot Caused by Fusarium oxysporum on Photinia × fraseri Dress in China.

Photinia × fraseri Dress was introduced to China in 1998 and has been widely used in gardens as an ornamental plant. From April 2021 to 2022, a diseasecausing blight and root discoloration in approximately 80% of P. × fraseri at several landscape sites in Xuanwu Lake Park. Symptomatic root tissues were immersed in 75% ethanol for 30 s followed by 1% NaClO for 90 s, rinsed with sterile water 3 times, and placed on Potato Dextrose Agar (PDA). After 3 days of dark incubation at 25 °C, white Fusarium-like colonies grew out from the symptomatic root tissue pieces. Three representative isolates (SG1, SG6, and SG23) were obtained and deposited in China's Forestry Culture Collection Center. The hyphae grew radially and the aerial hyphae were velvety, white or pinkish-white . After 20 days, macroconidia, microconidia, and chlamydospores were produced in the colonies on PDA. Macroconidia are sickle-shaped, slightly curved, 23-50.6 µm × 4-6 µm in size. Microconidia were numerous, oval or kidney-shaped, 6.7-12.6 µm × 3.5-5.7 µm in size, with germinating from one or both ends (Fig S1E, F). Chlamydospores were spherical, smooth, and round, in chains or solitary in hyphae (Fig S1G). All three isolates had identical morphological features. Phylogenetic analysis of concatenated CAMD, RPB2 and TEF1 sequences showed that the three isolates clustered in the same clade as F. oxysporum. Two-year old P. × fraseri potted seedlings (30-cm tall, n=12) were placed at the greenhouse (temperature; 26°C, daylight; 14 hours) for the pathogenicity tests . Roots of P. × fraseri were dipped in a 10 mL of the conidial suspension (106 conidia/mL) of each isolate for 2 hours, and the control plants were inoculated with sterile water. Results showed that after 21 days post-inoculation, all inoculated seedlings (n=9) showed crown and root rot . In contrast, none of the control seedlings (n=3) were affected. Re-isolation of three fungal isolates (infected root) showed that their morphology and gene markers sequence were identical to the original isolates thus fulfilled Koch's postulates. Globally, this is the first report of F. oxysporum causing crown blight and root rot of P. × fraseri, which is also a potential threat to the two parent hosts (P. serratifolia and P. glabra). Additional surveys are being conducted for mapping the distribution of F. oxysporum in the Nanjing Province of China.

First Report of Fusarium acuminatum Causing Dianthus chinensis root rot and foliage blight in China.

Dianthus chinensis is a popular ornamental plant that is widely cultivated in China. In May 2020, a disease was found at several landscape sites in Xuanwu District, Nanjing, China, causing symptoms of foliage blight and root discoloration on approximately 52% of one-year old D. chinensis plants. To recover the causal pathogen, samples of infected roots and leaves were cut into 5×5 mm2 pieces, surface-disinfected in 75% ethanol for 30 sec, followed by 1% NaClO for 90 sec, rinsed with sterile water three times and placed on potato dextrose agar (PDA) with 0.1 mg/mL of ampicillin at 25 °C. Hyphae growing on PDA were visible from both root and leaf tissues after three days. Individual hyphal tips were transferred to new PDA plates to obtain pure isolates. Three representative isolates were deposited in the China Forestry Culture Collection Center (CFCC 57545,57546, 57547). The hyphae grew radially, densely, and the aerial hyphae were velvety, white, yellow-white, or pink-white. Representative isolate Facu-DCY5 produced three types of conidia (microconidia, macroconidia, and chlamydospores). Macroconidia were sickle-shaped, measuring 25.7-55.4 µm × 3.2-4.6 µm (n=50). Microconidia were numerous, oval or kidney-shaped, measuring 6.8-11.9 µm × 3.5-4.8 µm (n=50). Conidia produced in the aerial mycelium were 16-34 × 2.2-5.3 µm (n=50). The ITS region, TEF1, calmodulin (CMDA), and RNA polymerase II second largest subunit (RPB2) were amplified with primers ITS1/ITS4, EF1/EF2, CL1/CL2A and 5F2/7CR , respectively and sequenced at Sangon Biotech (Nanjing, China). The ITS sequence of isolate Facu-DCY5 (GenBank No. ON307073.1) was identical to HQ165938.1, ON306850.1, OM964482.1. TEF1 (ON331997.1) was identical to LC546967.1, HQ165866.1, MZ158155.1. CMDA (ON331996.1) was identical to HQ412345.1, MZ921595.1 and MZ921597.1. RPB2 (ON331995.1) was identical to MZ997370.1. Maximum parsimony and maximum likelihood phylogenies of the Facu-DCY5 multilocus sequence data and those of several species within the F. tricinctum species complex identified the isolate from D. chinensis as F. acuminatum . Pathogenicity tests were performed using a conidial suspension (104 conidia/mL). Each plant (approx. 0.3 m in -height) was inoculated with 1 mL of the conidial suspension by mixing it into the potting soil (500 g). Control plants were treated with sterile distilled water. All inoculated plants (n=9) in three repeats of the assay exhibited foliage blight and root rot after 15 days, whereas all control plants (n=9) remained asymptomatic. Fusarium isolates with identical morphological features and molecular marker sequences to those of Facu-DCY5 were recovered from foliage blight and root tissues of all the inoculated plants. In China, F. acuminatum has been reported as a pathogen of Cucurbita maxima, Actinidia arguta, Polygonatum odoratumand Schisandra chinensis. This is the first report of F. acuminatum on D. chinensis in China. Considering the importance of D. chinensis to both ornamental nurseries and landscaping industries, we recommend that diseased plants be removed to prevent the spread of F. acuminatum, and that identification of the infecting isolates from D. chinensis at other sites and landscape locations be performed.

First Report of Crown and Root Rot Caused by Phytopythium helicoides on Photinia × fraseri Dress in China.

Photinia × fraseri Dress is a hybrid species of Rosaceae and Photinia genus which is widely cultivated in China. During 2020 and 2021, approximately 80% of plants growing in Xuanwu district of Nanjing, China, exhibited disease symptoms including blight, necrosis, and dieback of crowns and roots. Symptomatic root tissues collected from 2-year-old plants were rinsed with water, cut into 2-mm tissues which were surface-sterilized in 70% ethanol for 60 s, and plated onto 10% V8 PARP agar and incubated in the dark at 26°C for 3 days. Hyphae emerged from 70% of the samples. Two representative isolates (PF-he2, PF-he3) were obtained and deposited. Ten agar plugs (2×2 mm2) of each isolate were transferred into 10 mL of 10% V8 juice to produce mycelial mats. To stimulate sporangial production, 3-5 drops of soil extract solution (soil collected from healthy fields, immersed in sterile water, and filtered) were added to each plate. Sporangia were terminal, ovoid to globose or papillate. The zoospores were 7.1-9.3 µm in diameter. Each oogonium contained a single, smooth, spherical and aplerotic oospore with a diameter of 24.5-32.6 µm. These morphological properties resemble those of Phytopythium helicoides (CBS286.31 from S. F. Ashby). For molecular identification, the large subunit (LSU) rDNA, cytochrome c oxidase I (COXI) gene, and COXII gene were amplified using the primer pairs NL1/NL4 , FM55/FM52R , and FM66/FM58 . The LSU, COXI, and COXII sequences of isolate PF-he2 were 100% (763/763 nt), 98.07% (1066/1087 nt), 99.64% (561/563 nt) identical to isolate CBS 286.31 (AY598665.2), GDGJ6 (KT750956.1), and TC3 (MN952224.1), respectively. Based on the morphological and molecular analysis, the two isolates shared 100% homology were identified as Phytopythium helicoides. The pathogenicity of two isolates were tested on potted 2-yr-old (40-cm tall) P. × fraseri. Six plants were dug up to expose root balls which were wounded before inoculations with a sterile needle, and then inoculated with zoospore suspension (106 zoospore/mL). Controls were treated with ddH2O. Three seedlings/isolate were used for each treatment including controls. All plants were repotted using the original sterilized potting mix and pots. After inoculation, the plants were covered with plastic bags, and sterilized H2O was sprayed into the bags twice per day to maintain humidity and kept in a greenhouse at the day/night temperatures at 25/16 °C. All the inoculated plants showed lesions similar to those observed in the field after 23 days , whereas controls were asymptomatic. The isolates were reisolated from the lesions and sequenced as P. helicoides which has found causing root rot on Nelumbo nucifera, Rhododendron pulchrum, Zea mays in China, and also on Fragaria × ananassa in America, Peach Rootstock in California. Globally, this is the first report of P. helicoides causing crown blight and root rot of P. × fraseri. Management programs are under development to contain the spread of P. helicoides and treat diseased plants.