Lesion Margins Research Articles

First Report of Epicoccum sorghinum Causing Leaf Spot on Paeonia suffruticosa in China.

Tree peony (Paeonia suffruticosa Andr.; Paeoniaceae) is highly valued in Chinese culture for their ornamental and medicinal benefits in anti-inflammation, anti-arrhythmia, activating the immune system and protecting the cardiovascular system (Zhai et al. 2020). In May from 2019 to 2021, leaf spot symptoms were observed on tree peony in an area of 0.8 hm2 in Bozhou, Anhui Province, China; with a disease incidence of 35%. Initially, black-brown spots (0.5-2.4 mm) appeared on the leaves, as the disease developed, the spots enlarged to form round, black lesions. Small pieces of tissue taken from the margins of lesions were surface-disinfected by 75% ethanol for 20 s followed by 0.1% mercuric chloride for 3 min, rinsed with distilled sterile water for three times, placed on potato dextrose agar (PDA) at 25 ºC for 7 days in the dark.Fifteen single spore isolates were obtained as described (Dong 2009). When cultured on PDA for 10 days, colonies were white, villose, then became gray-brown, with the reverse side becoming reddish-brown; Chlamydospores globose or irregular shaped, dark brown, unicellular or multicellular (4.3 to 12.2 × 13.1 to 33.7 µm) (n = 200). Pycnidia were black-brown, mostly spheroid, and 86 to 128 × 110 to 197 µm (n = 50); Conidia were hyaline, ellipsoidal, unicellular, aseptate, and 4.1 to 5.2 × 1.8 to 2.4 µm (n = 200) . The DNA of a representative isolate AF21-1-2 was extracted using the CTAB protocol (Doyle and Doyle 1987). The rDNA internal transcribed spacer (ITS), β-tubulin (TUB) and 28s nrDNA (LSU) genes were amplified and sequenced using the primers ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass et al. 1995), and LR5/LR0R (Vilgalys and Hester, 1990), respectively (OK485136, OK631973, and ON358331). BLASTn analysis showed that the ITS, TUB, and LSU sequences had 99% (497/501 bp), 98% (530/539 bp), and 99% (878/882 bp) identity with the Epicoccum sorghinum isolates ESCZO20 (MN944541), CZ01 (MT254851) and Dl16-338 (LN907481), respectively. Maximum likelihood phylogenetic tree revealed that AF21-1-2 clustered with E. sorghinum isolates. For pathogenicity test, leaves of 3-year-old healthy tree peony plant in the field were inoculated with 105 conidia/ml spore suspension (prepared with 30-day-old cultures on PDA), added with 0.1% (v/v) Tween-80. Ten surface-sterilized leaves per plant were sprayed with 30 ml suspension until runoff, a total of 3 plants were inoculated. Leaves sprayed with sterilized water serve as a negative control. Each plant was covered with a plastic bag to maintain high relative humidity at 15 to 23°C and was monitored daily for symptom development. Typical symptoms appeared on more than 80% of the leaves after 7 days of inoculation, while control leaves remained symptomless. Cultures isolated from inoculated leaves had the same morphological and molecular traits as those that were previously used to test Koch's postulates. Based on the morphological and molecular characteristics, the causal agent on tree peony was identified as E. sorghinum (Aveskamp et al. 2010). To our knowledge, this is the first report of E. sorghinum causing leaf spot on tree peony in China. The disease will cause serious economic loss for tree peony if it is not managed properly. This report may provide the basis for diagnosis and control of the disease.

First report of Pythium aristosporum causing root rot on rice seedling in Taiwan.

Rice (Oryza sativa L.) is the principle staple crops in the World and its production can be severely damaged by Pythium species. Several Pythium species including P. afertile, P. arrhenomanes, P. dissotocum, P. elongatum, P. spinosum, have been recorded to cause rice seedling root rot in Taiwan (List of Plant Diseases in Taiwan edited by Tzean et al., 2019). During the survey of rice seedling diseases, we identified a new species of Pythium that causes seedling root rot on rice in commercial nursery trays in two nursery fields in 2019 in Taichung, Taiwan. Stunting and root rot symptom were found on the affected plants and up to 20% seedlings in a nursery tray showed similar symptoms. To isolate the pathogen, symptomatic roots were surface sterilized with 75% ethanol for 1 min and rinsed in sterile water. The margin of lesion was cut off, placed on 1.5% water agar and incubated at 28 ℃. After 24 h, the hyphal tips of a white colony growing from the diseased region were transferred to potato dextrose agar (PDA) medium. Koch's postulates were fulfilled by inoculating the germinated rice seeds with mycelia. Rice seeds of O. sativa var. Tainan11 (TN11) were treated with 75% ethanol and then 1.2% NaOCl for 15 min. The sterilized seeds were soaked in sterile water under dark condition for 3 days and the water was replaced every day. Five of the pre-germinated seeds with 2~5 mm embryonic shoot were placed in a sterile petri-dish and inoculated with 3-ml mycelial suspension (OD600 = 0.045) prepared by blending the mycelia of a 3-days PDA culture using an Oster 10 speed blender 6640 (Oster, USA). The seeds-mycelia were then covered with sterilized soil mixture of Akadama soil and rice husk (1:1, volume to volume) and incubated in a growth chamber at 28 ℃. Seven days post-inoculation, the inoculated seedlings showed stunting with short and necrotic roots (Fig. S1). The pathogen was reisolated from the diseased seedlings and identified with morphology and molecular methods. For morphological characterization, the pathogen was cultured on V8 agar to produce oogonia and zoospore (Chamswarng and Cook 1985). Globose oogonia with multiple antheridia (1-5 per oogonium), inflated filamentous sporangia, vesicle with abundant zoospores, main hypha with up to 6.57 μm wide and mature aplerotic oospores with diameter 24.35-30.81 μm (average= 27.22 μm; n=20) were observed (Fig. S1) that are similar to the descriptions for P. aristosporum (van der Plaats-Niterink 1981). Genomic DNA was extracted with CTAB method (Wang and White 1997) and the sequences of the internal transcribed spacer (ITS) region and gene region of β-tubulin (tub) and cytochrome c oxidase subunit II (cox II) were amplified with published primers (Villa et al., 2006). The obtained sequences were submitted to GenBank (accession nos: OL701302 (ITS), OL763269 (tub), and OL763270 (cox II); Fig. S2). Phylogenetic relationships between this Pythium pathogen and other 55 Pythium isolates, including the type species of P. aristosporum (ATCC11101), were conducted with the concatenated sequences of tub and cox II and analyzed by Bayesian interference (Fig. S3). Based on the tree built with tub and cox II sequences, this pathogen was identified as P. aristosporum that has not been reported in rice and other plants in Taiwan. It was observed in laboratory assays that this pathogen caused significant root-rot symptoms on several major rice varieties grown in Taiwan, including TN11, Tainung67 and Kaoshiung139. It may potentially cause severe crop loss in rice production, especially in nurseries. This identification provides important information on rice disease management.

First Report of Didymella rhei causing leaf spot on rhubarb in New York.

First Report of Didymella rhei causing leaf spot on rhubarb in New York E. J. Indermaur1, C. T. C. Day1, and C. D. Smart1† 1School of Integrative Plant Science, Section of Plant Pathology and Plant-Microbe Biology, Cornell University, Geneva NY 14456 †Corresponding author: C. D. Smart; Email: cds14@cornell.edu Rhubarb (Rheum spp.) is a perennial grown across the northern United States for petiole production (Foust & Marshall 1991). In August 2021, leaf spots were observed on rhubarb growing in a two-acre field in Erie Co., NY (Fig. S1). Approximately 30% of the plants in the field had leaf spot with disease severity of 5%. Initial symptoms on leaves were light brown, circular lesions with red margins that later coalesced into irregular spots. Lesion centers were dry with concentric rings, often perforating as they enlarged. Lesions on petioles were light brown, fusiform, and sunken with red margins. To identify the causal agent(s), symptomatic leaves and petioles from 50 plants (cultivar unknown) were collected with a W-shape sampling scheme. Lesion margins were surface sterilized with 70% ethanol for 60 s, 10% bleach for 60 s, rinsed in sterile water, plated on acidified potato dextrose agar (PDA), and incubated for two to four days at 20˚C. Hyphal tips from colony edges were transferred to new PDA plates. After 20 days, colonies (n=53) were olivaceous buff to grey olivaceous, producing white to grey, sparse aerial mycelium. Brown to black pycnidia were produced within five days in concentric rings around plate centers. Pycnidia were globose to subglobose, with one to two non-papillate or slightly papillate ostioles, and with mean diameter 75.8 (30.8 to 113.5) µm (n=20). Conidia were hyaline, ellipsoid or allantoid, and aseptate with mean ± SD dimensions of 6.2 ± 0.4 (4.9 to 8.1) x 2.2 ± 0.4 (1.3 to 3.3) µm (n=30) (Fig. S2). Based on these morphological characteristics, the isolates were initially identified as Didymella rhei [Ellis & Everh] (Qian Chen & L. Cai) (Boerema 2004). To confirm the identity, mycelia were scraped from PDA plates and homogenized using a TissueLyser II (Qiagen Inc.). Genomic DNA was extracted with a DNeasy Plant Mini Kit following manufacturer's instructions (Qiagen Inc.). PCR assays with primers ITS 4 and ITS 5 and fRPB2-7cR and RPB2-5F2 (Liu et al. 1999; Sung et al. 2007) were used to amplify the internal transcribed spacer (ITS) and the rpb2 gene regions of one representative isolate (strain RHU21204). Products were sequenced using Sanger chemistry. The sequences were deposited in GenBank with accession numbers OM903952 (ITS) and OM925897 (rpb2). The ITS and rpb2 sequences exhibited 99% (492/494 bp) and 100% (846/846 bp) identity with D. rhei accessions KF531831.1 and KP330428.1, respectively. Based on morphological and molecular characteristics, the pathogen was identified as D. rhei. To fulfill Koch's postulates, healthy leaves and petioles of four rhubarb seedlings (cultivars unknown) were spray-inoculated with a conidial suspension (1 × 107 conidia/ml) containing 0.2% Tween-20 from strain RHU21204. A tween suspension with no conidia was used as a control. Each treatment had three replicates. After inoculation, plants were placed in a 19˚C growth chamber with a 12-h photoperiod and misted for 30 min twice daily to maintain humidity above 80%. Initial symptoms were observed five days post inoculation (dpi), while control plants were asymptomatic. The pathogen was isolated 21 dpi from inoculated leaves and petioles with symptoms as described above (Fig. S1) and identified morphologically and molecularly as D. rhei. A representative isolate was deposited in the Cornell Plant Pathology Herbarium as CUP-070923. To our knowledge, this is the first report of D. rhei causing rhubarb leaf spot in New York and reducing the health and marketability of its host. Funding Source This project was funded by the College of Agriculture and Life Sciences, Cornell University. Literature Cited Boerema, G. H. et al. 2004. CABI Publishing. 288. Foust, C. M. and Marshall, D. E. 1991. HortScience 26:1360. DOI: 10.21273/HORTSCI.26.11.1360 Liu, Y. J. et al. 1999. Mol. Biol. Evol. 16:1799. Sung, G. H. et al. 2007. Mol. Phylogenet. Evol. 44:1204. DOI: 10.1016/j.ympev.2007.03.011.

The Role of Artificial Intelligence in Gastric Cancer: Surgical and Therapeutic Perspectives: A Comprehensive Review.

Stomach cancer has a high annual mortality rate worldwide necessitating early detection and accurate treatment. Even experienced specialists can make erroneous judgments based on several factors. Artificial intelligence (AI) technologies are being developed rapidly to assist in this field. Here, we aimed to determine how AI technology is used in gastric cancer diagnosis and analyze how it helps patients and surgeons. Early detection and correct treatment of early gastric cancer (EGC) can greatly increase survival rates. To determine this, it is important to accurately determine the diagnosis and depth of the lesion and the presence or absence of metastasis to the lymph nodes, and suggest an appropriate treatment method. The deep learning algorithm, which has learned gastric lesion endoscopyimages, morphological characteristics, and patient clinical information, detects gastric lesions with high accuracy, sensitivity, and specificity, and predicts morphological characteristics. Through this, AI assists the judgment of specialists to help select the correct treatment method among endoscopic procedures and radical resections and helps to predict the resection margins of lesions. Additionally, AI technology has increased the diagnostic rate of both relatively inexperienced and skilled endoscopic diagnosticians. However, there were limitations in the data used for learning, such as the amount of quantitatively insufficient data, retrospective study design, single-center design, and cases of non-various lesions. Nevertheless, this assisted endoscopic diagnosis technology that incorporates deep learning technology is sufficiently practical and future-oriented and can play an important role in suggesting accurate treatment plans to surgeons for resection of lesions in the treatment of EGC.

First report of Phytophthora pluvialis causing needle lesions on Pinus pinea in New Zealand

During routine surveillance in October 2020, needle lesions and necrosis were observed in two 42-year-old stone pines (Pinus pinea) situated in an amenity planting within an infected Pinus radiata plantation in Nelson, New Zealand. Symptoms were widespread across the foliage of infected trees and samples were sent to Scion's Forest Health Reference Laboratory. Approximately fifteen needles displaying yellow and red bands (Fig. 1) tested positive for Phytophthora using a Phytophthora ImmunoStrip® (Agdia, USA), and PCR and subsequent DNA sequencing with Ypap2F/Ypap2R primers (McDougal et al., 2021), indicated Phytophthora pluvialis. A Phytophthora species was isolated from diseased tissue. Morphological characteristics were consistent with those described by Reeser et al. (2013) for P. pluvialis. This identification was confirmed by sequencing the internal transcribed spacer region (White et al., 1990). The isolate was deposited in the National Forest Culture Collection (NZFS 5280) and symptomatic needles were lodged into the National Forest Mycological Herbarium (NZFRI 5931). Foliage symptoms caused by P. pluvialis on other conifers, including Pinus radiata, Pseudotsuga menziesii and Tsuga heterophylla, typically express as distinct khaki/red lesions and black resinous bands (Dick et al., 2014), rather than the yellow and red bands observed on the stone pines (Fig. 2). Pathogenicity testing was done by inoculating stone pine needles with P. pluvialis zoospores using a needle dip assay according to Dick et al. (2014). Stone pine needles from young healthy seedlings were placed in sterile pond water inoculated with P. pluvialis zoospores and incubated in darkness at 17°C for 24 hours. Control needles were placed in sterile pond water. Needles were then lain on moist paper towels in a sealed tray and incubated at 17°C at a 14-hour photoperiod. After ten days, olive/yellow lesions were observed on needles (Fig. 3). These lesions were lighter than those observed on field samples, however, this is common in detached needle assays of other species, such as P. pluvialis infection of Pinus radiata (Dick et al., 2014). Sporangia were observed near the lesion margin of needles (Fig. 4). Sections of symptomatic needles were surface sterilised and plated onto a Phytophthora-selective media (10% carrot agar + ampicillin, pimaricin, rifampicin, nystatin and hymexazol). The resultant cultures displayed morphological features of P. pluvialis, fulfilling Koch's postulates (Fig. 5). Control needles remained healthy (Fig. 3). To our knowledge, this is the first report of P. pluvialis causing needle disease on stone pine. Stone pine is an economically important species due to the production of high value pine nuts and timber. Although stone pine accounts for a small proportion (<1%) of New Zealand's forests, continued monitoring is recommended to assess the impact of disease on the growth and yield of these plantations. Forest Growers Levy Trust funded surveillance and diagnosis through the Forest Biosecurity Surveillance system.

Polarimetric Markers for Delineation of Lesional Margins during Brain Tumor Surgery

Polarimetric Markers for Delineation of Lesional Margins during Brain Tumor Surgery

First Report of Curvularia lunata Causing Leaf Spot on Oryza sativa in Sabah, Malaysian Borneo.

Rice (Oryza sativa L.) has been farmed in Malaysia since ancient times and is one of the most important commercial crops (Ma'arup et al. 2020). Throughout January to August 2022, chlorotic spots with brown halos ranging 2 to 10 mm wide were found on upper leaves of rice variety Mahsuri in the vegetative stage with a severity and incidence of approximately 60% and 100%, respectively in Kampung Tagas, Sabah, Malaysian Borneo (06°09'41.8"N, 116°13'45.1"E). As the disease developed, the spots coalesced into larger chlorotic spots. Three leaf pieces (5 x 5 mm) were excised from lesion margins, surface sterilized based on Khoo et al. (2022a), before plating on water agar (WA) at 25°C. Purification of fungi was conducted on WA using hyphal tip isolation. When three pure cultures were obtained, the fungi were cultured on potato dextrose agar (PDA) and WA for 7 days in 12 h light and 12 h dark at 25°C for the macro- and micro-morphological characterization, respectively. The colonies of the three isolates on PDA were initially gray, later becoming dark. Conidia (n=30) were fusiform, smooth-walled, dark-brown, and melanized with three transverse septa, measuring 7.3 to 11.4 × 16.2 to 27.2 µm. The isolates were named Tagas01, Tagas02, Tagas03. Genomic DNA was extracted from fresh mycelia of the pathogens based on the extraction method described by Khoo et al. (2022b). The primers ITS1/ITS4 (White et al. 1990), GPD1/GPD2 (Berbee et al. 1991), and EF1-983F/EF1-2218R (Schochet al. 2009) were used to amplify the internal transcribed spacer (ITS) region of rDNA, partial fragments of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and translation elongation factor (EF-1α) region, respectively based on PCR conditions as described previously (Khoo et al. 2022a). The sequences were deposited in GenBank under accession numbers OP268402, OP271304, OP271305 (677/677 bp) (ITS), OP270699, OP270703, OP270704 (609/613 bp) (GAPDH), OP270700-OP270702 (928/930 bp) (EF-1α). They were 99.35-100% similar to the Curvularia lunata ITS (HF934911), GAPDH (LT715821), and Curvularia dactyloctenicola EF-1α (MF490858) type sequences. Although C. dactyloctenicolais related to C. lunata, the conidia of the former are much wider making them easier to differentiate (Marin-Felix et al. 2017). Phylogenetic analysis using maximum likelihood based on the combined ITS, GAPDH and EF-1α sequences indicated that the isolate formed a supported clade to C. lunata. The pathogens were identified as C. lunata based on morphological and molecular characterization. Koch's postulates were performed. Three replicate healthy rice at the vegetative stage were sprayed with a spore suspension of 1 × 106 spore/ml in distilled sterilized water, prepared from 1-week-old fungal culture, grown in the dark on WA. Three replicate rice plants were sprayed with distilled sterilized water as control. Plants were covered with transparent polyethylene bags to keep moisture, and kept in a greenhouse at ~27°C. Bags were removed after 4 days of incubation. Monitoring and incubation were performed in greenhouse based on Iftikhar et al. (2022). The pathogenicity test was also performed using isolate Tagas02 and Tagas03. All inoculated leaves developed symptoms as described after 6 days post-inoculation, whereas no symptoms occurred on controls. The experiments were repeated twice. The reisolated fungi were identical to the pathogen morphologically and molecularly, thus fulfilling Koch's postulates. C. lunata has been reported in Peninsular Malaysia (Lee et al. 2012). This is the first report of C. lunata causing leaf spot on Oryza sativa in Sabah, Malaysian Borneo. This illness not only reduces yields and lowers milling quality, but it may also be mistaken for rice blast, necessitating needless fungicide spraying.

Gold versus ceramic - which will last longer for posterior indirect restorations?

Data sources Electronic search through PubMed (Medline), CENTRAL (Cochrane) and Embase databases in November 2021. Grey literature searches (OpenGrey, Pro-Quest, National research register and the register of clinical studies hosted by the US National Institute of Health).Study selection Randomised in vivo clinical trials (RCTs) with at least two-year follow-ups, that compared the longevity of adhesively luted indirect ceramic restorations against conventionally cemented indirect metal restorations. The teeth studies had to be about single tooth restorations, on premolars or molars, with sufficient information available for extraction; eg outcome assessment, reason for failure. No language or time restrictions were applied.Data extraction and synthesis Two authors performed study selection, data extraction and quality assessment independently and in duplicate. The following clinical data was extracted: number of participants and dental health status, location of the cavosurface margin, cavity type, reason for intervention, methods and materials for treatment, follow-up, lesion activity at moment of restoration placed, lesion extension (surfaces of restoration), lesion margin (enamel or dentine). It was decided if the restoration was renewed, repaired or re-cemented, then this was classed as outcome failure. Grading of evidence was performed according to the GRADE network.Results After duplicates were removed, 2,667 underwent title screening, 389 abstract screening, and 186 went on to have full text screening with 72 being excluded. In total, 110 eligible articles were then excluded from the 114 that were assessed leaving four RCTs meeting the eligibility criteria and being included in the synthesis, published between 2000 and 2013. Two studies were parallel-arm, the other two, split-mouth design. Overall, 443 restorations had been placed, 212 were adhesively luted ceramic, and 213 conventionally cemented metal restorations; these were all gold. The follow-up periods ranged from 5-7 years. Two studies compared inlays only, one looked at partial crowns only, and the fourth full crowns. The lowest failure rates were found for metal restorations, ranging from 0-11%. The failure rates for ceramic inlays and partial crowns were 5-9% over the same observation period. The failure rates for ceramic full crowns were 36% after five years. The most common failure for ceramic was found to be fracture of the restoration, whereas caries was the most frequent cause of failure for the gold restorations.Conclusions The risk of bias was high for all four studies and the grading of evidence showed a low level of evidence. Only one meta-analysis could be performed comparing conventionally cemented metal vs adhesively luted ceramic restorations, showing the relative risk to fail for conventionally cemented metal indirect restorations is significantly lower than adhesively luted ceramic indirect restorations.

First Report of Phytophthora inundata Associated with Decline and Death of Walnut (Juglans regia L.) in Italy.

Persian walnut (Juglans regia L.) is an important nut crop in Italy. In recent years, incidence of walnut decline and death has increased in many Italian commercial orchards. In early summer 2020, we observed a serious decline in approximately 5% of trees in a waterlogged area of a Veneto-region walnut orchard (J. regia, cv Lara). Symptoms included extensive foliar wilt and canopy decline associated with collar and root rot. Symptomatic tissues excised from larger roots of affected trees were surface disinfested for 1 min in a 1% NaOCl solution, rinsed for 5 min in sterile distilled water, and placed onto P5ARPH selective medium (Jeffers and Martin 1986). A Phytophthora-like organism was consistently isolated. Pure cultures were obtained by single-hyphal transfers onto potato dextrose agar (PDA). Isolates were identified as Phytophthora inundata based on morphological characteristics (Brasier et al. 2003), sequences of internal transcribed spacer (ITS) amplicons from universal primers ITS6 (Cooke et al. 2000) and ITS4 (White et al. 1990) and sequences of cytochrome c oxidase, subunit II (Cox II) from Fm75 and Fm78 primer pair (Martin and Tooley 2003). On carrot agar (CA), colonies had a characteristic stellate to broad lobed patterns. On this medium, optimal growth was at 28-30 °C (7,3 mm/day) and the upper temperature limit for mycelial growth was 37°C. Mycelial disks of isolate CREADC-Om306, grown on CA, were floated in Petri plates with soil extract solution and incubated under continuous fluorescent light at room temperature (25+/-2 °C). Within 48 to 72 h, sporangia were produced that were persistent, non-papillate, ovoid or ovoid-obpyriform, measuring 55.0 to 80.7 (length) x 41.3 to 65.2 (width) µm (averages 64.3+/-10.2 x 47.9+/-9.7 µm). Oospores and chlamydospores were absent. BLAST analysis of the amplicons from CREADC-Om306 revealed ITS sequences (854-bp; GenBank accession no. OK342200) and Cox II sequences (568-bp; GenBank accession no. OK349677) that shared 100% identity with published P. inundata sequences available in GenBank (acc. n. AF266791 for ITS; MT458994 for Cox II). Pathogenicity tests were conducted in the greenhouse on six 2-year-old walnut (J. regia, cv Lara) plants. Four of the plants were inoculated with CREADC-Om306 on two opposite sides of each plant's stem at 1-2 cm above soil line. A cork borer was used to remove a 5-mm disk of bark that was replaced by a 5-mm diameter mycelial plug from 10-day-old cultures of the pathogen on PDA. Two control plants were treated in the same way except the bark wounds were inoculated with sterile PDA plugs. Plants were kept in greenhouse at 24 ± 2°C. After 3 months, lesions had developed from all points of inoculation with. P. inundata (mean lesion length 55,25+/-6,22 mm) and the pathogen was reisolated from the lesion margins of all inoculated plants. The control plants remained symptomless and did not yield the pathogen. P. inundata is widely distributed across the world as a plant pathogen on several native as well as horticultural crops, especially in riparian or other areas subject to flooding or waterlogging. This report is the first to document P. inundata as a pathogen on Persian walnut and adds it to the diverse list of known susceptible perennial native, ornamental, and agricultural hosts of this organism. In addition to P. inundata, which belongs to the major Phytophthora ITS Clade 6, other members of the clade including P. megasperma (Belisario et al. 2012) and P. gonapodyides (Belisario et al. 2016) have been described as walnut pathogens.

Risks and treatment for recurrent intraepithelial cervical lesions

Background. Persistently high incidence of cervical cancer in Russia and significant number of cases detected in the late stages necessitate the improvement of secondary prophylaxis of this disorder.Aim. To assess risk factors for recurrent high-grade cervical intraepithelial neoplasia (CIN2+) (high grade squamous intraepithelial lesions, HSIL) after cervical conization.Materials and methods. This study included 62 patients with recurrent HSIL treated in Novosibirsk Regional Clinical Oncology Dispensary, E. N. Meshalkin National Medical Research Center, “Zdorovye” LLC, “Avismed” LLC, Tomsk National Research Medical Center of the Russian Academy of Sciences, and Federal Research and Clinical Center for Specialized Medical Care and Medical Technologies, Federal Biomedical Agency of the Russian Federation in 2017–2021. We analyzed patients’ human papillomavirus (HPV) status, performed repeated examination of excised tissue specimens to evaluate the severity of lesions and resection margins, as well as immunohistochemical examinations. We found that mean time to cytologically confirmed recurrent HSIL was 16.0 ± 5.6 months. All patients were HPV-positive. Repeated histological examination demonstrated that 18 samples had positive resection margins or endocervical crypt involv ement. Fifty-seven samples had positive staining for p16 at immunohistochemical examination; 46 samples had Ki-67 &gt;30 %, which indicated high risk of recurrence. Treatment of patients with recurrent HSIL included repeated excision up to healthy cervical tissues, followed by intravaginal therapy with Cervicon-DIM 100 mg twice a day (for 3 months). Follow-up examinations after 18.0 ± 6.2 months on average showed no HPV persistence and no HSIL recurrence.Conclusion. Endocervical crypt involvement along the primary resection margin, underestimated severity and depth of lesions (at the first surgery), and persistence of HPV infection are the main risk factors for recurrent cervical dysplasia or carcinoma in situ. Combination treatment that includes additional excision with a subsequent course of Cervicon-DIM is sufficient and effective.

Oesophagectomy within 30 days after noncurative endoscopic resection for oesophageal cancer: are we able to follow this 'golden' interval?

Endoscopic mucosal resection (EMR) is considered the first-line treatment for early-stage intramucosal lesion pT1a (m1-3) and carefully selected pT1b (sm1) non-circumferential oesophageal carcinomas [1]. This endoscopic treatment strategy has great advantages compared to surgical oesophageal resection, with a reduced invasiveness, decreasing morbidity and mortality rates and allowing the organ preservation with subsequent improvement in patients’ quality of life. Moreover, recent data demonstrated that EMR has equivalent long-term outcomes when compared to both oesophagectomy for intramucosal tumour (m1-3) or limited to the invasion of few microns of the muscularis mucosa with favourable histopathological factors (sm1), with an acceptable low risk of positive lymph node (LN) occurring in <5% of cases [2]. However, there are many circumstances in which EMR has to be deemed as noncurative, leading to consider radical oesophagectomy because of the high risk of recurrence or LN metastases. These circumstances are: tumour size >2 cm, invasion >sm1, excavated or polypoid lesion, circumferential lesion, positive lateral and/or vertical margin, poorly differentiated tumour, evidence of lymphovascular or perineural invasion and suspicion of positive LN [3]. If oesophagectomy is the recommended treatment option for noncurative EMR, the question that remains unclear is the optimal interval between the 2 procedures regarding the surgical risk if the delay is too short due to post-resection inflammation, and the risk of a pathological upstaging if the delay is too long due to local disease progression.

First Report of Flower Stalk Wilting Caused by Rhizopus oryzae on Chinese cabbage in China.

Chinese cabbage (Brassica rapa L. ssp. pekinensis) is one of the main biennial vegetables in China and its flowers can be produced in the second year. In May 2021, approximately 50% of the flower stalks of Chinese cabbage wilted in a field in Laizhou, China. Water-soaked lesions were first observed on the lateral shoots of flower stalks, leading to wilting at a later stage. Small diseased tissues were excised from the margin of lesions, surface disinfected in 75% alcohol, rinsed in distilled water twice, and transferred onto potato dextrose agar (PDA) medium for incubation at 28 ℃. Five fungal isolates were obtained using single spore isolation method. The fungal colonies were initially white and became gray or black within 5 days. The columella was globose to subglobose and 82.86±5.25 μm (n=5) in diameter; sporangiophores were smooth-walled, simple or branched; the globose sporangia were 86.06±15.37 μm (n=5) in diameter and black; the sporangiospores were subglobose and abundant and 5.23±0.98 μm (n=5) in diameter; and the rhizoids were dark brown and 5.69±1.82 μm (n=5) wide. A cetyl tri-methyl ammonium bromide method was used to extract DNA from 3-day-old hyphae (Ausubel et al. 1987). PCR was performed for ITS (White et al. 1990), the RNA polymerase II large subunit (RPB1) gene (Voigt et al. 2000) and the actin (ACT) gene (Stiller et al. 1997). The DNA sequences of the five isolates were identical, therefore, the sequence of Isolate RO21 was submitted to GenBank. According to BLAST search, the ITS (MZ452687), RPB1 (OK431470), and ACT (OK431471) sequences showed 99.66% similarity to Rhizopus oryzae Strain CBS 112.07 (NR103595), 100% to Strain CBS 127.08 (KJ566325) and 100% to Strain CBS 102660 (KJ551423), respectively. A neighbor-joining phylogenetic tree was reconstructed based on the ITS of Isolate RO21 and 14 other Rhizopus species sequences obtained from GenBank. Isolate RO21 was found to be most closely related to R. oryzae and far from other species. Based on morphological and phylogenetic characteristics, Isolate RO21 was identified as R. oryzae (Dolatabadi et al. 2014, Kwon et al. 2015, Palemón-Alberto et al. 2020). Sporangiospores were harvested from 5-day-old PDA cultures, suspended in sterilized distilled water, adjusted to 106 spores/ml and amended with 0.1% Tween-80. Chinese cabbage inbred line "A54-1" was inoculated near the middle of the flower stalk by applying 20 μl of spore suspension (106 spores/ml) to each of three sites wounded using a sterilized knife or to the unwounded site. Sterilized distilled water was used as the control. Forty flower stalks (20 for the inoculation treatment and the rest for the control) selected from ten plants were used for pathogenicity test. All plants were incubated in a growth chamber at 28/22 °C (day/night), with 80 to 90% of relative humidity. Wilting symptoms similar to those in the field were observed in the wounded flower stalks after 5 days and in the non-wounded flower stalks after 15 days. All control flower stalks remained asymptomatic. The fungus was re-isolated from the artificially infected flower stalks and identified as R. oryzae by morphological characteristics and sequencing to fulfill the Koch's postulates. To our knowledge, this is the first report that R. oryzae causes flower stalk wilting on Chinese cabbage in China. The results can provide the basis for future studies on the occurrence, prevention and management of this disease.

Occurrence of Leaf Spot Caused by Diaporthe phaseolorum on Bletilla striata in China.

Bletilla striata (Thunb.) Rchb. f. (Orchidaceae) is a traditional Chinese medicinal plant widely distributed in eastern and southern Asia. In April of 2020, a leaf spot disease on B. striata was observed in plant nurseries in Guilin, Guangxi Province, China. Disease incidence was estimated at approximately 20% (n = 150 plants) across the survey area (~ 0.3 ha). The initial symptoms were small, reddish to brown spots, circular or irregular in shape. Subsequently, they developed into large dark brown, irregular lesions. As the lesions coalesced, leaves withered and defoliated. To isolate the causal agent, eighteen small pieces (~ 5 mm2) were collected from the margin of the necrotic lesions on Chinese ground orchid, surface disinfected (2 min in 1% NaOCl, and rinsed three times in sterile water), and placed on potato dextrose agar (PDA) at 26°C for 3 days. Hyphal tips were transferred to PDA to obtain pure cultures. Twelve isolates were obtained, of which eight isolates had similar morphological characteristics. After 7 days growth on PDA, colonies were grayish-white, fluffy, with white aerial mycelium. After 3 weeks, colonies formed white aerial mycelial mats, and pycnidia developed. The α-conidia were abundant, hyaline, aseptate, ellipsoidal to fusiform, measuring 4.6 to 6.7 μm × 2.1 to 3.0 μm (n = 55), whereas the β-conidia were hyaline, long, slender, straight or curved, measuring 10.3 to 17.2 μm × 0.9 to 1.8 μm (n = 59). Morphological features were similar to Diaporthe sp. (Santos et al. 2011, Udayanga et al. 2015). For further molecular identification, DNA was extracted from the mycelia of the representative isolate BJ26.3 following the CTAB (cetyltrimethylammonium bromide) method (Guo et al. 2000). The internal transcribed spacer (ITS) region, partial translational elongation factor subunit 1-α (EF-1α), calmodulin (CAL) , histone H3 (HIS3), β-tubulin (TUB) genes were amplified and sequenced using the primer pairs ITS1/ITS4, EF1-728F/EF1-986R, CAL-228F/CAL-737R, CYLH3F/H3-1b, and Bt2a/Bt2b, respectively (White et al. 1990, Guarnaccia et al. 2018). The obtained sequences were deposited in NCBI GenBank under the following accession numbers: OK560457, OK539595, OK539592, OK506726, OK539598. BLAST analysis of the deposited sequences showed 99 to 100% identity with accession numbers KC343177 (563/566 bp), KC343903 (521/523 bp), KC343419 (423/427 bp), KC343661 (340/340 bp), KC344145 (658/662 bp) of D. phaseolorum CBS 127465 (Guarnaccia et al. 2018). In addition, a phylogenetic analysis using concatenated sequences confirmed BJ26.3 as D. phaseolorum. Furthermore, pathogenicity tests were carried out on 1.5-year-old B. striata plants. Healthy leaves on three plants (1 leaf per plant) were inoculated with 5 × 5 mm mycelial discs of strain BJ-26.3 from 3-day-old PDA cultures. Another three plants treated with sterile water served as the control. All plants were covered with transparent plastic bags and maintained in a greenhouse at 26°C with a 12 h photoperiod. Nine days post-inoculation, the inoculated leaves showed leaf spot symptoms, while the control plants remained healthy. The experiments repeated three times showed similar results. Finally, D. phaseolorum was consistently re-isolated from the infected leaves and confirmed by morphology and sequencing, fulfilling Koch's postulates. To our knowledge, this is the first report of D. phaseolorum causing leaf spot of B. striata worldwide. This study might provide important information for growers to manage this disease.

Pseudomonas cichorii causing leaf spot disease on Banxia (Pinellia ternata) in China.

Banxia (Pinellia ternata) is an important Chinese medicinal material in the family Araceae and is a widely grown herb in China. In September 2021, a leaf spot disease was observed on Banxia field, with an incidence rate of 35 to 40 % in a 4-ha field, in Zhongxiang City, Hubei Province of China. Symptoms were observed as yellow-white centers, water-soaked edges, irregular lesions, and gradually developed into a yellowish-brown center and a dark-brown edge. Necrotic spots gradually increased, leading to leaf chlorosis and plant death. Margins of leaf lesions were excised form diseased tissue and were plated on nutrient agar (NA) using serial dilution. Growth on NA was predominantly cream-colored circular bacterial colonies with undulated margins. Characterization of three randomly chosen bacterial isolates (JYB1, JYB7 and JYB8) suggests they are Gram-negative, levan negative, arginine dihydrolase negative, oxidase positive, potato soft rot positive, and tobacco hypersensitive positive. Isolates were identified as Pseudomonas cichorii based on the LOPAT scheme (Cottyn et al. 2009). Taxonomic positioning was confirmed genetically by PCR analysis using primers set: 16S rRNA gene universal primers 27F/1492R (Weisburg et al. 1991) and hrcRST gene specific primers Hcr1/Hcr2 (Cottyn et al. 2011). Homology search of 16S rRNA gene sequences (GenBank accessions: JYB1, MZ749668; JYB7, MZ823822; and JYB8, MZ823823) indicated 99.93 % (1396 bp) identity with P. cichorii strains (GenBank accessions: MK356431, JX913785, MZ723344). Similarly, comparison of the hrcRST locus (GenBank accessions: JYB1, MZ977010; JYB7, MZ977011; and JYB8, MZ977012) shared 99.38% (812 bp) with P. cichorii strains (GenBank accessions: CP007039, CP074349, GU324131). Koch's postulate was performed on healthy 30-day-old Banxia plants to confirm pathogenicity of the isolated strains. Leaves were injected with 50 μL bacterial suspensions (1x108 cfu/ml) by sterile syringe. The negative control was inoculated with sterile water. The inoculated Banxia plants were incubated at 28 °C, 70 to 80 % relative humidity, and exhibited water-soaked lesions on the leaf surface within two days around the inoculation sites. Within seven days, all leaves withered and plants died. In contrast, control plants remained healthy and symptomless. The pathogen was consistently reisolated from diseased plants and morphologically and molecularly identified as P. cichorii, while no bacterial colonies were isolated from the control plants, fulfilling Koch's postulates. To our knowledge, this is the first report of bacterial leaf spot caused by P. cichorii on Banxia in China. As one of the main producing areas of Banxia in China, Jianghan Plain of Hubei Province has a planting area of nearly 20 square kilometers. The occurrence of this bacterial disease has the potential threat to the Banxia industry, more research is needed for breeding disease resistance and for developing chemical control.

Mediating effect assessment of ifosfamide on limb salvage rate in osteosarcoma: A study from a single center in China

Osteosarcoma is one of the most prevalent primary bone malignancies in children and adolescents. Surgery and chemotherapy are the standard treatment methods of osteosarcoma. Methotrexate, adriamycin, and cisplatin, and methotrexate, adriamycin, cisplatin, and ifosfamide regimens are both first-line neoadjuvant chemotherapy regimens for osteosarcoma. Moreover, the use of ifosfamide is highly controversial. Most studies of ifosfamide focused on the overall survival rate and event-free survival rate; few studies concentrated on surgical options. We conducted this retrospective study to compare the baseline characteristic of amputation and limb salvage osteosarcoma patients. Furthermore, we analyzed the direct and indirect roles in surgical decision-making and found that ifosfamide may play a partial mediating role in the surgery option choice by mediating tumor mass volume change, tumor response, and the shortest distance from the center of main blood vessels to the margin of the tumor lesion.

ICG Enema to Make Rectal Shaving Easier and Safer

Study Objective To use ICG enema to improve the surgical quality during rectal shaving. Design Surgical video. Setting Local GYN MIS hospital, single surgeon. Patients or Participants Patients with solitary rectal DIE nodule. Interventions After complete adhesiolysis and excision of all other pelvic endometriosis lesions except the rectal nodule, 12.5 mg ICG was diluted in 60 cc normal saline. The diluted ICG solution was injected into rectal lumen through anus and was retained in rectum (ICG enema) during the whole rectal shaving procedure. Measurements and Main Results After ICG enema, we started the rectal shaving procedure, carefully mapped the lesion margin, cut into rectal muscular layer with monopolar scissors under pure cutting mode. The submucosa layer can be easily recognized by the ICG green color. As long as we identified the green submucosa layer, we can easily stay precisely between the muscle layer and submucosa layer, keep the green submucosa and mucosa layer intact, with minimal risk of enterotomy. After rectal shaving, the muscular defect of the rectal wall can also be clearly identified by the exposed green submucosa layer. Rectal wall repair can then be confidently performed, transversely re-approximating the seromuscular layer, embedding all the green submucosa layer. Conclusion By ICG enema, staining the mucosa and submucosa layer with green ICG color, we can easily identify the submucosa layer during the rectal shaving procedure. The risk of enterotomy, and incomplete rectal wall repair after shaving can both be minimized. To use ICG enema to improve the surgical quality during rectal shaving. Surgical video. Local GYN MIS hospital, single surgeon. Patients with solitary rectal DIE nodule. After complete adhesiolysis and excision of all other pelvic endometriosis lesions except the rectal nodule, 12.5 mg ICG was diluted in 60 cc normal saline. The diluted ICG solution was injected into rectal lumen through anus and was retained in rectum (ICG enema) during the whole rectal shaving procedure. After ICG enema, we started the rectal shaving procedure, carefully mapped the lesion margin, cut into rectal muscular layer with monopolar scissors under pure cutting mode. The submucosa layer can be easily recognized by the ICG green color. As long as we identified the green submucosa layer, we can easily stay precisely between the muscle layer and submucosa layer, keep the green submucosa and mucosa layer intact, with minimal risk of enterotomy. After rectal shaving, the muscular defect of the rectal wall can also be clearly identified by the exposed green submucosa layer. Rectal wall repair can then be confidently performed, transversely re-approximating the seromuscular layer, embedding all the green submucosa layer. By ICG enema, staining the mucosa and submucosa layer with green ICG color, we can easily identify the submucosa layer during the rectal shaving procedure. The risk of enterotomy, and incomplete rectal wall repair after shaving can both be minimized.

First report of Calonectria canadiana causing peach fruit rot in China.

Peach (Prunus persica (L.) Batsch) is one of the most popular fruits grown in Northern China. In July 2021, a fruit rot outbreak on the peach cultivar "Yonglian Sweet" occurred after unusual rains in Baoding, Hebei Province, China. Sixty peach trees from three orchards were assessed, and a 30% disease incidence was estimated. The disease initiated as a small concave spot on the fruit surface expanding circularly rotting the fruit (3-5 cm deep) with the appearance of grayish-white mycelia (Figure S1A). The infected fruit did not disintegrate but turned light brown. To identify the pathogen, 20 infected fruits were collected, and fruit tissues from lesion margins were inoculated on the potato dextrose agar (PDA) medium. A total of 15 fungal pure cultures with highly similar morphological characteristics were obtained by the hyphal-tipping method. The fungal culture formed smooth-edged colonies of extensive, dense, wooly aerial mycelium, with color changing from sienna to luteous, and to grayish-white along the radius of colonies (Figure S1B) Chlamydospores were extensive and developed micro-sclerotia after 20 d of growth. The conidiophore produced three branches in a "broom" shape, with the primary branch ranging 7.5-25.0 μm in length, the secondary branch 5.5-15.5 μm, and the tertiary branch 10-12.5 μm (N = 30). The top of the tertiary branch tapered and produced conidia. Conidia were colorless and culm-like, 40.0-57.5 μm long and 3.8-6.25 μm wide (N = 30). Hyphae occasionally produced spherical chlamydospores with a diameter of around 7.5 μm (N = 30). Conidia germinated after 12 h in moist conditions, and germ tubes originated from multiple points on the conidia. Based on these morphological features, the isolated fungus was identified as Calonectria spp. (Lombard et al. 2010). Six loci, including ITS, act, cmdA, his3, tef1, and tub2, were amplified and sequenced for molecular identification of an isolate F099 using primers listed in Table S1. The obtained ITS (528 bp, GenBank accession no. OL635556), act (263 bp, OL694221), cmdA (470 bp, OL694222), his3 (432 bp, OL694223), tef1 (487 bp, OL694224), and tub2 (535 bp, OL694225) sequences showed 100% similarity to the ex-type strain of Calonectria canadiana, CMW 23673 (accession nos. MT359667, MT334976, MT335206, MT335446, MT412737, and MT412958, respectively; Figure S1D) (Kang et al. 2001, Lechat et al. 2010, Liu et al. 2020). The isolate F099 of C. canadiana was further subjected to pathogenicity tests. Koch's postulates were performed by placing three mycelial disks (ten-day old, 5 mm) with conidia on the sterile needle-acupunctured surface of healthy fruits of the peach cultivar "Yonglian Sweet" (N= 10). Mock inoculations with sterile PDA disks were served as a control. All the inoculated fruits were kept in a moist chamber (25℃, 16-h light and 8-h dark period). The inoculation assay was repeated twice. Rotting symptoms developed on all the inoculated fruits about 5 days post-inoculation (dpi) and grayish-white mycelia appeared around ten days post inoculation while mock inoculated fruits did not show any rotting. The pathogen of interest was re-isolated from the inoculated fruits and validated as C. canadiana by ITS and tef1 sequences. All above evidence collectively indicates that the fungal pathogen causing the peach fruit rot is C. canadiana. The new host plant and new geographic distribution reported here will inform future management of this fungal species.

First Report of Alternaria alternata Causing Leaf Spot Disease on Daylily in China.

Daylilies (Hemerocallis spp.; Xanthorrhoeaceae) originated from Eastern Asia and are widely cultivated as perennial ornamentals from the tropics to their native high latitudes. In June 2021, daylily cultivar 'Tao Hua Zhai' with leaf spot symptoms were found at the Shanghai Institute of Technology, Shanghai, China. The disease prevalence was about 14.5 % in a 33,000 m2 planting area indicated by survey statistics. Symptoms of the disease initially appeared as small, circular, brown spots on the leaves. As disease progressed, spots increased gradually until they were distributed uniformly over the lamina, the leaf tip became withered and the rest of the leaf became chlorotic. Symptomatic leaf tissue pieces (5 × 5 mm) from lesion margins were sterilized with 75 % ethanol for 1 min, rinsed three times with sterile distilled water, then incubated on potato dextrose agar (PDA) plates at 28 °C in the dark. A pure culture (ATHF-1) was obtained. Its upper surface on PDA was olive green with loose aerial hyphae, and its lower surface was brown.Conidiophores were brown, single or branched, producing numerous short chains conidia. Conidia were obclavate to obpyriform or ellipsoid, pale brown to dark brown, with a short cylindrical beak at the tip, contained 2-6 transverse septa and 0-4 longitudinal septa. The size of conidia were 15.9-47.3 µm × 7.6-16.6 µm (n=50), and length/width ratios were 1.51 to 4.92. Based on the morphological characteristics, the fungus was identified as Alternaria spp. (Simmons, 2007). For molecular characterization, three genes (the internal transcribed spacers [ITS], plasma membrane ATPase [ATPase] and major allergen Alt a 1) of ATHF-1 were amplified with primer pairs ITS1/ITS4 (White et al. 1990), ATPDF1/ATPDR1 (Lawrence et al. 2013) and Alt-for/Alt-rev (Hong et al. 2005), respectively. The sequences were deposited in GenBank (ITS, MZ983611; ATPase, MZ962978; Alt a 1, OK021654). Blastn searches showed the nucleotide sequences of ATHF-1 were highly similar to the reference sequences of Alternaria tenuissima (ITS, 99 % to KU982591; ATPase, 98 % to MT833928; Alt a 1, 100 % to MT109294). A phylogenetic tree based on the ITS, ATPase and Alt a 1 sequences was constructed by MEGA7.0, which showed that ATHF-1 was closely related to A. tenuissima and A. alternata. But according to Woudenberg et al. (2015), they were synonymized under the species name A. alternata. So, based on morphological and molecular characteristics, the fungus was identified as A. alternata. For pathogenicity tests, ten healthy two-month-old potted seedlings from tissue culture daylilies were sprayed with 20 ml of suspension (approximately 2×105 spores/ml), ten daylilies were used as controls and sprayed with sterile water. After covering with transparent plastic bags for 48 h to maintain humidity, the plants were placed in the greenhouse at 25 ℃ with 12 h photoperiod. The pathogenicity tests were repeated twice. Seven days after inoculation, lesions appeared on the plants inoculated with the pathogen, which were consistent with the symptoms observed in the field, while the controls remained symptomless. The morphological characteristics and gene sequences of the re-isolated strain from the diseased leaves were consistent with those of the inoculated strain. To our knowledge, this is the first report of A. alternata affecting leaf spot disease on daylily in China. Identification of the causal agent of the disease is important for developing effective disease management strategies.

Artificial intelligence for detecting and delineating a small flat-type early esophageal squamous cell carcinoma under multimodal imaging.

Early diagnosis of esophageal squamous cell carcinoma (ESCC) is crucial for improving its prognosis. Effective endoscopic biopsy is key to ensuring no missed diagnoses or misdiagnosis of lesions occur and accurate delineation of the margins of lesions is essential for effective biopsies. Recently, the application of artificial intelligence (AI) in the endoscopic diagnosis of early ESCC has increased; however, most AI systems identified early ESCC with rough square frames [1] [2] [3]. For the purpose of precise lesion segmentation, we have developed a new AI system, based on a state-of-the-art algorithm called the YOLACT model [4], that integrates multiple endoscopic imaging modalities to detect and delineate the margins of early ESCC in real time.

Red Foliage Blight of × Taxodiomeria peizhongii Caused by Neopestalotiopsis clavispora Newly Reported in China.

× Taxodiomeria peizhongii Z.J. Ye, J.J. Zhang & S.H. Pan is a hybrid of Taxodium mucronatum and Cryptomeria fortunei. It can adapt to various site conditions and has a good saline-alkali tolerance, which is a unique tree species in eastern China. In August 2020, a red foliage blight with an incidence of 70% (105/150 plants) was found on the leaves of × T. peizhongii in a nursery, Shanghai, China (121°21'12"E, 31°41'56"N). It developed from apical leaves of branches downwards. The infected leaves became reddish brown and withered. Fresh specimens were collected from 3 infected trees. Small samples (3-4 mm2) from lesion margins were sterilized, plated on potato dextrose agar (PDA) and incubated at 25°C. Nine isolates of the same fungus were obtained. Three representative isolates (DFS1-3, DFS1-8, and DFS1-9) were used for morphological and molecular studies and deposited in the China's Forestry Culture Collection Center (cfcc57401 to cfcc57403). The colonies of three isolates on PDA grew fast, covering the entire plate with white cottony mycelia in 7 days. Acervuli of DFS1-3 were 618-996 × 586-945 µm (n = 50). Conidiogenous cells were 4.4-9.8 µm (n = 50) long. Conidia were 5-celled, clavate to fusiform, smooth, 19-24 × 6.4-8.8 µm (n = 50). The 3 median cells were dark brown to olivaceous, central cell was darker than other 2 cells, and the basal and apical cells were hyaline. All conidia developed one basal appendage (3.4-8 µm long; n = 50), and 2-3 apical appendages (15-30 µm long; n = 50), filiform. The morphological characters of DFS1-8 and DFS1-9 were almost identical to DFS1-3. Based on morphological studies, the isolates were Neopestalotiopsis sp.. The DNA of 3 isolates was extracted. The internal transcribed spacer region (ITS), β-tubulin 2 (TUB2) and translation elongation factor 1-alpha (TEF1-α) loci were amplified using the primer pairs ITS1/ITS4, T1/Bt-2b, EF1-728F/EF-2. BLAST result showed that ITS of the three isolates were identical to Pestalotiopsis sp. at a high level (greater than 99%), and TUB2, TEF1-α were highly similar with Neopestalotiopsis sp. (greater than 99%). The sequences were deposited in GenBank [Accession Nos. OM188301 and OM222696 to OM222697 for DFS1-3; OM188303 and OM222698 to OM222699 for DFS1-8; OM188302 and OM222700 to OM222701 for DFS1-9]. A maximum likelihood and Bayesian posterior probability analyses using IQtree v. 1.6.8 and Mr. Bayes v. 3.2.6 with the concatenated sequences (ITS, TUB2, TEF1-α) clustered 3 isolates together with N. clavispora including type isolate (MFLUCC 12-0281). Based on the morphology and phylogeny, the fungus was N. clavispora. To confirm pathogenicity, 9 healthy 2-yr-old seedlings, and 10 leaves per seedling were wounded with a sterile needle and inoculated with conidial suspension (106 conidia/mL). Three control plants were sprayed with sterile water. Seedlings were covered with plastic bags after inoculation and kept in a greenhouse at 25 ± 2°C and RH 80%. Seven days after inoculation, all inoculated leaves were reddish brown and withered like those observed in the field, whereas the control plants remained symptomless. N. clavispora was successfully reisolated from the infected tissues. This pathogen has been reported to cause leaf blight on many other hosts, such as Ligustrum lucidum and macadamia, but in recent years, the disease has also been reported on flowers, such as Anthurium. It has not been reported on Taxodium and Cryptomeria. This is the first report of N. clavispora infecting × T. peizhongii in the world. These data will help select appropriate fungicides for managing this newly emerging disease.