Zinc finger transcription factor ZFP1 is associated with growth, conidiation, osmoregulation, and virulence in the Polygonatum kingianum pathogen Fusarium oxysporum.

Rhizome rot is a destructive soil-borne disease of Polygonatum kingianum and adversely affects the yield and sustenance of the plant. Understanding how the causal fungus Fusarium oxysporum infects P. kingianum may suggest effective control measures against rhizome rot. In germinating conidia of infectious F. oxysporum, expression of the zinc finger transcription factor gene Zfp1, consisting of two C2H2 motifs, was up-regulated. To characterize the critical role of ZFP1, we generated independent deletion mutants (zfp1) and complemented one mutant with a transgenic copy of ZFP1 (zfp1 tZFP1). Mycelial growth and conidial production of zfp1 were slower than those of wild type (ZFP1) and zfp1 tZFP1. Additionally, a reduced inhibition of growth suggested zfp1 was less sensitive to conditions promoting cell wall and osmotic stresses than ZFP1 and zfp1 tZFP1. Furthermore pathogenicity tests suggested a critical role for growth of zfp1 in infected leaves and rhizomes of P. kingianum. Thus ZFP1 is important for mycelial growth, conidiation, osmoregulation, and pathogenicity in P. kingianum.

Seizure, Motor, and Cognitive Outcomes After Epilepsy Surgery for Patients With Sturge-Weber Syndrome: Results From a Multicenter Study.

BACKGROUND AND OBJECTIVES: Surgery is widely performed for refractory epilepsy in patients with Sturge-Weber syndrome (SWS), but reports on its effectiveness are limited. This study aimed to analyze seizure, motor, and cognitive outcomes of surgery in these patients and to identify factors associated with the outcomes. METHODS: This was a multicenter retrospective observational study using data from patients with SWS and refractory epilepsy who underwent epilepsy surgery between 2000 and 2020 at 16 centers throughout China. Longitudinal postoperative seizures were classified by Engel class, and Engel class I was regarded as seizure-free outcome. Functional (motor and cognitive) outcomes were evaluated using the SWS neurologic score, and improved or unchanged scores between baseline and follow-up were considered to have stable outcomes. Outcomes were analyzed using Kaplan-Meier analyses. Multivariate Cox regression was used to identify factors associated with outcomes. RESULTS: A total of 214 patients with a median age of 2.0 (interquartile range 1.2-4.6) years underwent surgery (focal resection, FR [n = 87]; hemisphere surgery, HS [n = 127]) and completed a median of 3.5 (1.7-5.0) years of follow-up. The overall estimated probability for being seizure-free postoperatively at 1, 2, and 5 years was 86.9% (95% CI 82.5-91.6), 81.4% (95% CI 76.1-87.1), and 70.7% (95% CI 63.3-79.0), respectively. The overall estimated probability of being motor stable at the same time post operatively was 65.4% (95% CI 58.4-71.2), 80.2% (95% CI 73.8-85.0), and 85.7% (95% CI 79.5-90.1), respectively. The overall probability for being cognition stable at 1, 2, and 5 years was 80.8% (95% CI 74.8-85.5), 85.1% (95% CI 79.3-89.2), and 89.5% (95% CI 83.8-93.2), respectively. Both FR and HS were effective at ensuring seizure control. For different HS techniques, modified hemispherotomy had comparable outcomes but improved safety compared with anatomical hemispherectomy. Regarding FR, partial resection (adjusted hazard ratio [aHR] 11.50, 95% CI 4.44-29.76), acute postoperative seizure (APOS, within 30 days of surgery; aHR 10.33, 95% CI 3.94-27.12), and generalized seizure (aHR 3.09, 95% CI 1.37-6.94) were associated with seizure persistence. For HS, seizure persistence was associated with APOS (aHR 27.61, 9.92-76.89), generalized seizure (aHR 7.95, 2.74-23.05), seizure frequency ≥30 times/month (aHR 4.76, 1.27-17.87), and surgical age ≥2 years (aHR 3.78, 1.51-9.47); motor stability was associated with severe motor defects (aHR 5.23, 2.27-12.05) and postoperative seizure-free status (aHR 3.09, 1.49-6.45); and cognition stability was associated with postoperative seizure-free status (aHR 2.84, 1.39-5.78) and surgical age <2 years (aHR 1.76, 1.13-2.75). DISCUSSION: FR is a valid option for refractory epilepsy in patients with SWS and has similar outcomes to those of HS, with less morbidity associated with refractory epilepsy. Early surgical treatment (under the age of 2 years) leads to better outcomes after HS, but there is insufficient evidence that surgical age affects FR outcomes. These findings warrant future prospective multicenter cohorts with international cooperation and prolonged follow-up in better exploring more precise outcomes and developing prognostic predictive models. CLASSIFICATION OF EVIDENCE: This study provides Class IV evidence that in children with SWS and refractory seizures, surgical resection-focal, hemispherectomy, or modified hemispherotomy-leads to improved outcomes.

CDBA: a novel multi-branch feature fusion model for EEG-based emotion recognition.

EEG-based emotion recognition through artificial intelligence is one of the major areas of biomedical and machine learning, which plays a key role in understanding brain activity and developing decision-making systems. However, the traditional EEG-based emotion recognition is a single feature input mode, which cannot obtain multiple feature information, and cannot meet the requirements of intelligent and high real-time brain computer interface. And because the EEG signal is nonlinear, the traditional methods of time domain or frequency domain are not suitable. In this paper, a CNN-DSC-Bi-LSTM-Attention (CDBA) model based on EEG signals for automatic emotion recognition is presented, which contains three feature-extracted channels. The normalized EEG signals are used as an input, the feature of which is extracted by multi-branching and then concatenated, and each channel feature weight is assigned through the attention mechanism layer. Finally, Softmax was used to classify EEG signals. To evaluate the performance of the proposed CDBA model, experiments were performed on SEED and DREAMER datasets, separately. The validation experimental results show that the proposed CDBA model is effective in classifying EEG emotions. For triple-category (positive, neutral and negative) and four-category (happiness, sadness, fear and neutrality), the classification accuracies were respectively 99.44% and 99.99% on SEED datasets. For five classification (Valence 1-Valence 5) on DREAMER datasets, the accuracy is 84.49%. To further verify and evaluate the model accuracy and credibility, the multi-classification experiments based on ten-fold cross-validation were conducted, the elevation indexes of which are all higher than other models. The results show that the multi-branch feature fusion deep learning model based on attention mechanism has strong fitting and generalization ability and can solve nonlinear modeling problems, so it is an effective emotion recognition method. Therefore, it is helpful to the diagnosis and treatment of nervous system diseases, and it is expected to be applied to emotion-based brain computer interface systems.

First Report of Root Rot on Paris polyphylla var. yunnanensis caused by Fusarium solani in Yunnan, China.

Paris polyphylla var. yunnanensis is a perennial herb with diverse chemical components having wide-ranging pharmacological effects. The demand for P. polyphylla var. yunnanensis as a raw material increases greatly and currently exceeds 1,000 tons per year (Zhou et al. 2021). In September 2021, root rot was observed on P. polyphylla var. yunnanensis in Mangshi, Yunnan province, China. Average disease incidences in the fields reached 15%, with diseased plants exhibiting yellowing and wilting leaves, as well as browning and rotting roots. Cross sections (5 × 5 mm2) cut from the margin of symptomatic and asymptomatic root tissues were surface-sterilized for 30 s with 75% ethanol, followed by 180 s with 1% sodium hypochlorite. After rinsing thrice with sterile distilled water, the fragments were transferred to potato dextrose agar (PDA) plates and incubated at 28°C in the dark. Ten isolates were obtained, and single spore isolation was performed. These isolates showed similar morphological characters, with colonies ranging in color from white to pale cream and sparse mycelia. Conidia were produced on the top or side of phialides. Microconidia were oval or reniform, 0- or 1-septate, with a diameter of 5.1-10.7 µm × 1.6-3.9 µm (average 7.6 µm × 2.8 µm) (n=30). The macroconidia were straight to slightly curved or sickle-shaped, 3- to 5-septate, with a diameter of 15.1-27.9 µm × 2.8-4.0 µm (average 21.0 µm × 3.6 µm). Chlamydospores were smooth, nearly round, and 3.3-6.6 (average 4.9) µm in diameter. Genomic DNA were extracted from mycelia of the two isolates. The nuclear ribosomal internal transcribed spacer (ITS), translation elongation factor 1 alpha (EF1α), and the second largest subunit of nuclear DNA-directed RNA polymerase II (RPB2) were amplified with the primer pairs of ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and fRPB2-5F/fRPB2-7cR (Liu et al. 1999), respectively. These two isolates exhibited the same nucleotide sequences (ITS, OP646781; EF1α, OP661172; RPB2, OP661173), with BLASTn analyses showing 100%, 99.66%, and 99.65% identity, respectively, with Fusarium solani (syn. Neocosmospora solani) (Crespo et al. 2019) strain NRRL 43474 (ITS, EF453097; EF1α, EF452945; RPB2, EF469984). A phylogenetic tree was constructed using MEGAX based on the nucleotide sequences of ITS, EF1α, and RPB2, using the maximum likelihood method. The isolate was classified into the F. solani clade. According to the morphology and sequence analyses, the isolate was identified as F. solani (Chehri et al. 2015), and named PpFs1. To test the pathogenicity of the isolate PpFs1, the roots of four years old P. polyphylla var. yunnanensis plants were dipped in 107 spore/mL suspension filtered from potato dextrose broth (PDB) for 30 min, while control roots were dipped in sterile water. After inoculation, all plants were transplanted in pots filled with sterile soil and kept at 25°C with a 12/12-h light/darkness photoperiod. Six plants were used for each treatment, and repeated thrice. Two months after inoculation, the infected plants showed wilted leaves and rotted roots, while controls remained asymptomatic. PpFs1, identified by morphology and ITS, was re-isolated from infected plants, and was found to comply with Koch's postulates. To the best of our knowledge, F. oxysporum and F. concentricum causes Paris polyphylla var. Chinensis stem rot in China. But this is the first report of root rot on P. polyphylla var. yunnanensis being caused by F. solani in Yunnan, China.

First report of Didymella curtisii causing leaf scorch on spider lily (Hymenocallis littoralis) in China.

Hymenocallis littoralis (Jacq.) Salisb, commonly known as spider lily, is a Amaryllidaceae species widely cultivated in southern China for ornamental and medicinal purposes (Anusha et al., 2016). In 2020, there was a devastating outbreak of leaf scorch of H. littoralis from July to September in Kunming city, Yunnan province of China (E102.8268°, N24.8371°), with 97% disease incidence. The initial spots were small and reddish-brown but gradually expanded to large irregular lesions with yellow centers. The leaves then turned yellow and withered from tip towards the petiole in the severely infected plants. For identification, leaf sections (5 × 5 mm2) cut from the margins of the lesions were surface-sterilized for 3 min with 1% sodium hypochlorite, rinsed three times with sterile distilled water, placed on potato dextrose agar (PDA), and incubated at 28°C at a 12-h photoperiod for three days. One fungus was isolated from 90% of the samples, of which three monosporic isolates were selected using a method of agar dilution lineation separation. After 4 days of incubation, colonies were white-yellowish, and changed to beige-mustard after 7 days. The pycnidia were produced after 10 days incubation on oatmeal agar (OA), and they were black-brown, subglobose, and ostiolate. Chlamydospores mainly formed in the aerial mycelia, globose, often in chains, brown or pale. The conidia were ellipsoidal or clavate, (0-)1-3-septate, 2.1 to 10.8 × 1.0 to 3.2 µm (n=60). The total genomic DNA of three isolates was extracted from mycelia. The nuclear ribosomal internal transcribed spacer region (ITS), the second largest subunit of nuclear DNA-dependent RNA polymerase II (rpb2), the 28S nuclear ribosomal large subunit rRNA gene (LSU), and beta-tubulin gene (tub2) were amplified using the ITS1/ITS4 (White et al., 1990), fRPB2-5F/fRPB2-7cR (Liu et al., 1999), LR0R/LR5 (Schoch et al., 2012), and Btub2Fd/Btub4Rd (Woudenberg et al., 2009) primer pairs, respectively. The amplicons were cloned in the pMD19-T vector (Code No. 6013, Takara, Kusatsu, Japan) and sequenced bi-directionally. These three isolates had the same nucleotide sequences, one of which was submitted to NCBI (ITS, OM279485; rpb2, OM304305; LSU, OP800249; tub2, OQ108870). BLASTn analyses showed that ITS, rpb2, LSU, and tub2 were genetically 100%, 98.49%, 99.89%, and 97.89%, respectively, identical with MN973518, MT018130, MN943724, and MT005618 genes of Didymella curtisii strain CBS 288.29. Phylogenetic tree was constructed with MEGAX based on the nucleotide sequences of ITS, rpb2, LSU, and tub2 using the maximum likelihood method. The fungus isolated from diseased leaves of H. littoralis was grouped into the same clade with D. curtisii. According to the morphology and sequence analyses, the isolate was D. curtisii (Chen et al., 2015) and was named isolate HlDc1. To confirm the causal agent of the disease, a spore suspension with 106 spores of HlDc1/mL was smeared on healthy leaves of six months old with brushes. Leaves in the control group were smeared with sterile water. All inoculated plants were incubated at 28°C under a 12-h photoperiod in a moisture chamber. The pathogenicity tests were conducted three times with six plants each time. Fifteen days post-inoculation, the leaves inoculated with HlDc1 developed red-brown lesions, whereas the control leaves remained asymptomatic. Isolate HlDc1 was re-isolated from infected leaves. To our knowledge, this is the first report of leaf scorch on H. littoralis in Yunnan province, China, caused by D. curtisii. The results laid the foundation for epidemiological forecasting and scientific control of this disease.