Complete mitochondrial genomic sequence of Auricularia delicata (Auriculariaceae), an edible Chinese mushroom.

Auricularia delicata (Mont.) Henn. 1893 is an edible and medicinal jelly mushroom popular in China. Here, we report the assembly and annotation of a complete A. delicata mitochondrial genome based on data sequenced using an Illumina NovaSeq 6000 platform. The length of the complete circular A. delicata mitochondrial genome is 189,696 bp, with a GC content of 34.1%. The A. delicata mitochondrial genome contains 60 genes, including 32 protein-coding genes, 26 tRNA genes, and two rRNA genes. Phylogenetic analysis indicated that A. delicata clustered with the Auricularia group, alongside A. auricula-judae and A. heimuer. Additionally, A. delicata was found to be genetically distant from other species of Polyporales, Russulales, and Agaricales. This genome will provide an invaluable reference for the continued study and utilization of A. delicata and other Auricularia species.

Identification of Colletotrichum aenigma as the new causal agent of leaf blight disease on Aucuba japonica Thunb., and screenings of effective fungicides for its sustainable management.

Aucuba japonica Thunb is an evergreen woody ornamental plant with significant economic and ecological values. It also produces aucubin, showing a variety of biological activities. It is widely planted in the southwest region of China, including karst landscape areas in Guizhou Province. In January 2022, a serious leaf blight disease was observed on the leaves of A. japonica in the outdoor gardens of Guizhou University, Guiyang, Guizhou, China. The causal agent was identified as Colletotrichum aenigma through amplification and sequencing of the internal transcribed spacer (ITS) region, translation of the chitin synthase (CHS) and actin (ACT) genes, and morphological characterizations. Koch's postulates were confirmed by its pathogenicity on healthy leaves, including re-isolation and identification. To our knowledge, this is the first report of C. aenigma causing leaf blight on A. japonica worldwide. To identify pathogen characteristics that could be utilized for future disease management, the effects of temperature and light on mycelial growth, conidia production, and conidial germination, and the effects of humidity on conidial germination were studied. Optimal temperatures for mycelial growth of C. aenigma BY827 were 25-30°C, while 15°C and 35°C were favorable for conidia production. Concurrently, alternating 10-h light and 14-h dark, proved to be beneficial for mycelial growth and conidial germination. Additionally, conidial germination was enhanced at 90% humidity. In vitro screenings of ten chemical pesticides to assess their efficacy in suppressing C. aenigma representative strain BY827. Among them, difenoconazole showed the best inhibition rate, with an EC50 (concentration for 50% of maximal effect) value of 0.0148 µg/ml. Subsequently, field experiment results showed that difenoconazole had the highest control efficiency on A. japonica leaf blight (the decreasing rate of disease incidence and decreasing rate of disease index were 44.60 and 47.75%, respectively). Interestingly, we discovered that C. aenigma BY827 may develop resistance to mancozeb, which is not reported yet among Colletotrichum spp. strains. In conclusion, our study provided new insights into the causal agent of A. japonica leaf blight, and the effective fungicides evaluated provided an important basis and potential resource for the sustainable control of A. japonica leaf blight caused by C. aenigma in the field.

Plant Growth Promotion and Biocontrol of Leaf Blight Caused by Nigrospora sphaerica on Passion Fruit by Endophytic Bacillus subtilis Strain GUCC4.

Passion fruit (Passiflora edulis Sims) is widely cultivated in tropic and sub-tropic regions for the production of fruit, flowers, cosmetics, and for pharmacological applications. Its high economic, nutritional, and medical values elicit the market demand, and the growing areas are rapidly increasing. Leaf blight caused by Nigrospora sphaerica is a new and emerging disease of passion fruit in Guizhou, in southwest China, where the unique karst mountainous landscape and climate conditions are considered potential areas of expansion for passion fruit production. Bacillus species are the most common biocontrol and plant-growth-promotion bacteria (PGPB) resources in agricultural systems. However, little is known about the endophytic existence of Bacillus spp. in the passion fruit phyllosphere as well as their potential as biocontrol agents and PGPB. In this study, 44 endophytic strains were isolated from 15 healthy passion fruit leaves, obtained from Guangxi province, China. Through purification and molecular identification, 42 of the isolates were ascribed to Bacillus species. Their inhibitory activity against N. sphaerica was tested in vitro. Eleven endophytic Bacillus spp. strains inhibited the pathogen by >65%. All of them produced biocontrol- and plant-growth-promotion-related metabolites, including indole-3-acetic acid (IAA), protease, cellulase, phosphatase, and solubilized phosphate. Furthermore, the plant growth promotion traits of the above 11 endophytic Bacillus strains were tested on passion fruit seedlings. One isolate, coded B. subtilis GUCC4, significantly increased passion fruit stem diameter, plant height, leaf length, leaf surface, fresh weight, and dry weight. In addition, B. subtilis GUCC4 reduced the proline content, which indicated its potential to positively regulate passion fruit biochemical properties and resulted in plant growth promotion effects. Finally, the biocontrol efficiencies of B. subtilis GUCC4 against N. sphaerica were determined in vivo under greenhouse conditions. Similarly to the fungicide mancozeb and to a commercial B. subtilis-based biofungicide, B. subtilis GUCC4 significantly reduced disease severity. These results suggest that B. subtilis GUCC4 has great potential as a biological control agent and as PGPB on passion fruit.

The complete mitochondrial genome of the edible mushroom Pleurotus giganteus (Agaricales, Pleurotus) and insights into its phylogeny.

Pleurotus giganteus (Berk.) Karunarathna & K.D. Hyde 2011 is one of the largest edible mushrooms integrating medicinal value and edible value. The complete mitochondrial genome of the edible fungus P. giganteus was published in this paper. It was determined using Pacbio and Illumina sequencing. The circular molecule is 102,950 bp in length, consisting of 30 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, and 24 transfer RNA (tRNA) genes. The base composition of the whole mitogenome is A (37.3%), T (37.7%), G (12.2%), and C (12.8%). The phylogenetic tree shows P. giganteus was the basal taxon in Pleurotus and closely related to Pleurotus citrinopileatus Singer 1990.

Detection of a spinning object with circular procession using an optical vortex beam.

The rotational Doppler effect (RDE) provides an efficient way to measure rotational frequency using an optical vortex beam. Crucially, most research based on the RDE just involves a spinning object or a spinning object coupled with a longitudinal velocity along the beam propagation. We analyze the interaction mechanism between optical orbital angular momentum and a spinning object with circular procession and experimentally demonstrate simultaneous measurements of two rotational frequencies. This technique broadens application of the RDE in optical metrology and remote detection of targets with micro-motions.

First report of green mold disease caused by Penicillium citrinum on Dictyophora rubrovalvata in China.

Dictyophora rubrovolvata is a saprophytic mushroom widely cultivated in China, including Guizhou Province for its high nutritional, medicinal, and economical values (Chen et al. 2021). In May 2021, green mold disease was observed on the fruiting bodies of D. rubrovolvata, causing its death or preventing it from forming a sporocarp, in an indoor-production facility at Asuo village, Baiyun District Guiyang city, Guizhou Province, China (26°73'51" N, 106°72'88" E). The disease incidence was 60%-70% in the affected 1.33-ha growing area, causing a serious economic loss. To identify the causal agent, a total of 15 samples with symptomatic symptoms were collected. Small pieces (5 mm × 5 mm) were cut from the diseased tissues, surface sterilized in 0.4% NaClO for 5 min, washed three times with sterilized water, placed on potato dextrose agar (PDA) medium, and incubated at 24 °C for 7 days. Twenty-one pure cultures were obtained by single-spore isolation method. The colonies were initially white but after seven days as conidia developed they turned green. Hyphae were hyaline and guttulate. Conidiophores were verrucose stipes, triverticulate, and phialides flask shaped. Conidia were smooth and pale green, with subglobose to globose shape measuring 2.0-2.5 × 1.8-2.5 µm (n=50). Based on these morphological characteristics, the isolates matched the description of the genus Penicillium (Visagie et al. 2014). To confirm the identity, DNA of five representative isolates (QS001, QS005, QS008, QS015, QS017) was extracted according to the manufacturer's instructions (Biomiga Fungal DNA Extraction Kit; CA, USA). Afterwards, PCR was performed to amplify ITS region, calmodulin and ß-tubulin genes using primer pairs ITS1/ITS4 (White et al. 1990), CMD5/CMD6 (Glass et al. 1995), and Bt2a/Bt2b (Hong et al. 2006), respectively. BLASTN analysis of these sequences showed the best matches with Penicillium citrinum CBS 139.45 (ITS region: 98.60% (493/500 bp) identity to accession MH856132.1; CMD: 99.79% (469/470 bp) identity to accession MN969245.1; ß-tubulin:100% (407/407 bp) identity to accession GU944545.1). Representative sequences of the sequenced DNA regions were deposited in GenBank (ITS region: OK446552; CMD: OK492612; ß-tubulin: OK482677). Furthermore, a phylogenetic tree was constructed with MEGA 7 based on the concatenated sequences. Koch's postulates were met to confirm the pathogenicity of the representative isolate (QS001) on D. rubrovolvata. Six discs (5mm×5mm) from actively growing P. citrinum QS001 colonies (5-day-old) were placed on six fruiting bodies of D. rubrovolvata (5-month-old). Mock inoculations were performed using PDA discs only without any fungus. The inoculation sites were wrapped with a sterilized 200-µm nylon mesh. All fruiting bodies were incubated at 23°C ± 2°C under a 0-h/24-h photoperiod and 80% relative humidity (RH) after inoculation. After 14 days, green mold was observed on all P. citrinum QS001 inoculated mushrooms. In contrast, no disease was observed in mock inoculated group. The disease assays were repeated three times. P. citrinum QS001 was isolated from all inoculated D. rubrovolvata and verified via the molecular analysis mentioned above. To the best of our knowledge, this is the first report that P. citrinum causes green mold on D. rubrovalvata in China and further studies should focus on managing this disease to prevent any disease outbreaks.

Wheat embryo globulin nutrients ameliorate d-galactose and aluminum chloride-induced cognitive impairment in rats.

Wheat embryo globulin nutrient (WEGN), with wheat embryo globulin (WEG) as the main functional component, is a nutritional combination that specifically targets memory impairment. In this study, we explored the protective role of WEGN on Alzheimer's disease (AD)-triggered cognitive impairment, neuronal injury, oxidative stress, and acetylcholine system disorder. Specifically, we established an AD model via administration of d-galactose (d-gal) and Aluminum chloride (AlCl3) for 70 days, then on the 36th day, administered animals in the donepezil and WEGN (300, 600, and 900 mg/kg) groups with drugs by gavage for 35 days. Learning and memory ability of the treated rats was tested using the Morris water maze (MWM) and novel object recognition (NOR) test, while pathological changes and neuronal death in their hippocampus CA1 were detected via HE staining and Nissl staining. Moreover, we determined antioxidant enzymes by measuring levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) in serum, cortex, and hippocampus, whereas changes in the acetylcholine system were determined by evaluating choline acetyltransferase (ChAT), and acetylcholinesterase (AChE) activities, as well as choline acetylcholine (Ach) content. Results revealed that rats in the WEGN group exhibited significantly lower escape latency, as well as a significantly higher number of targeted crossings and longer residence times in the target quadrant, relative to those in the model group. Notably, rats in the WEGN group spent more time exploring new objects and exhibited lower damage to their hippocampus neuron, had improved learning and memory activity, as well as reversed histological alterations, relative to those in the model group. Meanwhile, biochemical examinations revealed that rats in the WEGN group had significantly lower MDA levels and AChE activities, but significantly higher GSH, SOD, and ChAT activities, as well as Ach content, relative to those in the model group. Overall, these findings indicate that WEGN exerts protective effects on cognitive impairment, neuronal damage, oxidative stress, and choline function in AD rats treated by d-gal/AlCl3.

First Report of Fruit rot Caused by Phytophthora nicotianae on Passion Fruit in Guangxi Province, China.

Passion fruit (Passiflora edulis) is an economically important fruit crop in many tropical and subtropical regions worldwide. In recent years, passion fruit was widely cultivated in Guangxi Province. In 2020, a rot disease occurred on immature fruit of passion fruit in several commercial orchards of Nanning, Guangxi, caused about 50% incidence. The first appeared as small, irregular, water-soaked, brown lesions on immature fruit. As the disease progressed, the lesions rapidly enlarged, causing fruit rot. A layer of sparse white mycelia appeared on the lesions at high humidity. The disease first developed in June, its peak periods from August to September. Five diseased fruits were collected from five different orchards. The edges of symptomatic fleshy mesocarp tissue were cut into pieces (5 mm × 5 mm), surface-sterilized in 75% ethanol solution for 60 s, rinsed three times with sterilized distilled water, and plated on potato dextrose agar (PDA). Plates were incubated at 25°C in the dark. After 5 days, similar white colonies with abundant aerial mycelia developed from all plated tissue samples. Five isolates were obtained, and they were identified as Phytophthora nicotianae based on morphological characteristics and DNA analysis. Spherical hyphal swellings were commonly produced. Numerous sporangia were formed in sterile soil extract. Sporangia were ovoid or obpyriform, papillate, and measured 25 to 58 µm (average 41 µm) × 21 to 45 µm (average 29 µm). Chlamydospores were spherical and 19 to 43 µm in diameter (average 30 µm) (Erwin and Ribeiro 1996). The genomic DNA of a representative isolate Seg2-5 was extracted from mycelia through modified CTAB method (Murray and Thompson 1980). The rDNA internal transcribed spacer (ITS) region, ypt1, and coxII were amplified and sequenced with primers ITS1/ITS4 (White et al., 1990), Yph1F/Yph2R (Schena et al. 2008), and FM75F/FM78R (Villa et al. 2006), respectively. BLAST searches of the ITS, ypt1, and coxII sequences (Accession No. MW470847, MW770870, and MW770871) showed 99 to 100% identity with sequences of P. nicotianae (Accession No. JF792540, MK058408, and MH551183). Based on morphological characteristics and phylogenetic analysis, isolate Seg2-5 was identified as P. nicotianae. To confirm pathogenicity, asymptomatic and immature fruits 'Mantianxing' of passion fruit were previously disinfested in 0.5% sodium hypochlorite. Mycelial plugs of isolate Seg2-5 were placed onto the surface of fruits by nonwounded and pin-prick inoculation. Blank plugs were used as negative controls. Each treatment had five replicates and the test was repeated twice. Fruits were maintained in plastic boxes at 28°C and the initial disease spots appeared at 3 dpi or 5 dpi with wounded or non-wounded inoculation. After 7 to 10 days, all inoculated fruits showed similar symptoms as observed initially in the field, whereas control fruits remained healthy. P. nicotianae was successfully reisolated and identified from the inoculated fruits based on morphological characters and ITS sequence, thus confirming Koch's postulates. P. nicotianae had been previously isolated from passion fruit in South Africa (Van and Huller 1970), Vietnam (Nguyen et al. 2015), and Fujian Province of China (Luo et al. 1993). To our knowledge, this is the first report of P. nicotianae infecting passion fruit in Guangxi Province, China.

Analysis on the expression and value of CCL2 and CCL3 in patients with osteoarthritis.

In this study, we investigated the clinical value of CC chemokine ligand 2 (CCL2) and CC chemokine ligand 3 (CCL3) in osteoarthritis (OA). A prospective analysis was performed on 126 patients with rheumatoid arthritis of the knee (observation group (OG)) who admitted to Qilu hospital from August 2016 to August 2018 and 135 healthy controls (control group (CG)) who underwent physical examinations during the same period. The concentrations of CCL2 and CCL3 in peripheral blood were compared between the two groups, and the predictive value of the two on the occurrence, efficacy and prognosis of recurrence of OA were analyzed. In addition, an OA rat model was established to detect the relative protein levels of CCL2 and CCL3 in rat knee joint tissues. There were no statistically significant differences between the two groups in baseline data such as age, body mass index (BMI), gender, smoking, drinking, educational level, family medical history and living environment (P > 0.05). CCL2 and CCL3 increased in peripheral blood of patients in the OG (P < 0.05), both of which were positively correlated with rheumatoid factor (RF) (P < 0.001). CCL2 and CCL3 were of good predictive value for the occurrence, efficacy and prognosis of recurrence of OA (P < 0.001). The relative protein levels of CCL2 and CCL3 in bone and joint tissues of OA rats were significantly higher than those of normal rats (P < 0.001). CCL2 and CCL3 are elevated in peripheral blood of OA patients, which have good predictive value for the occurrence, efficacy and prognosis of recurrence of OA, indicating their potential roles as excellent markers for diagnosis and treatment of OA in the future.

First report of Cedecea neteri Causing Yellow rot Disease in Pleurotus pulmonarius in China.

Pleurotus pulmonarius is a popular edible fungus and widely cultivated in many areas of China. In June 2018, yellow rot (more than 10% incidence) was found on the first crop of P. pulmonarius fruiting bodies in a mushroom factory in Nanning, Guangxi Province, China. At first, yellow water-soaked lesions appeared in the infected fruiting bodies. Lesions then spread and purulent tissues were formed. Severe rot induced production of deformed fruiting bodies and offensive odor. Internal sections of the diseased tissue (approximately 0.5 × 0.5 cm) were sterilized in 75% alcohol for 30 s, rinsed three times with sterilized and deionized water, crushed and suspended in sterilized and deionized water. The suspension was spread on the Luria-Bertani (LB) medium. After incubation at 30°C for 2 days, dominant bacterial colonies were oyster white, smooth, convex, and circular. Individual colonies were transferred two times to LB medium using the conventional streak plate techniques to obtain the pure cultures. The cells were gram-negative, short rods, motile, and no capsules or endospores were observed. Using a BoJian Gram-negative bacteria biochemical analysis kit (5 CARDS, Hopebio, Qingdao, China), data were obtained and analyzed, showing that the isolated strain belongs to the Cedecea genus (positive for ß-galactosidase, citric acid, arginine, sucrose, mannitol, sorbitol, D-glucose, gelatin hydrolysis and VP test but negative for H2S, urease, oxidase, indole, rhamnose, melibiose, amygdalin, lysine, ornithine, lactose, inositol and arabinose). Amplified 16S rDNA gene sequences (1,424 bp, GenBank accession No. MT925570) of the isolate using the universal primers 27f and 1492r (Lane 1991) exhibited 99.86% identity with Cedecea neteri M006 (CP009458.1). Based on its morphological characteristics, 16S rDNA sequences, and biochemical test results, the strain was identified as C. neteri. Pathogenicity tests for this strain were performed with bacterial suspensions (approximately 1 × 108 CFU/ml) after growing for 24 h in LB medium at 30°C. Mycelia of P. pulmonarius were cultivated for 60 days in plastic bags. Then young fruiting bodies were formed after induced with low temperature stimulation to serve as a host source. The prepared bacterial suspensions were directly sprayed onto the surface of three bags of fruiting bodies; another three bags were sprayed with sterilized and deionized water as negative control. All inoculated fruiting bodies were then incubated at 20°C with 90 to 95% relative humidity. All experiments were repeated three times. After 2 days, all the fruiting bodies inoculated with the bacterial suspensions showed yellow water-soaked lesions, and the normal growth of the fruiting bodies was inhibited. An offensive odor then developed along with a severe soft rot that was similar to the disease symptoms observed under natural conditions. The fruiting bodies of negative control were growing healthily with no symptoms. Koch's postulates were fulfilled by isolating bacteria from lesions on artificially inoculated fruiting bodies that were identical to the original isolates based on morphological characteristics, 16S rDNA sequences and biochemical test results. C. neteri was formally reported as a pathogen to humans that could cause bacteremia (Farmer et al. 1982). Recently, it has also been reported causing soft rot disease on mushrooms of Pholiota nameko (Yan et al. 2018) and yellow sticky disease on mushrooms of Flammulina velutipes (Yan et al. 2019). However, to the best of our knowledge, this is the first report of C. neteri-induced yellow rot disease of P. pulmonarius in China.

Upregulation of circular RNA circ_0034642 indicates unfavorable prognosis in glioma and facilitates cell proliferation and invasion via the miR-1205/BATF3 axis.

Accumulating studies demonstrate the critical role of circular RNAs (circRNAs) in the pathogenesis of various types of cancers. Previously, hsa_circ_0034642 has been found elevated in glioma tissues compared with the normal tissues, as proved by high-throughput microarray. We further investigated its expression level in 52 paired tissues and different glioma cell lines by quantitative reverse transcription-polymerase chain reaction. In addition, Fisher's exact test, Kaplan-Meier curves, and Cox analyses were performed to elucidate the clinical significance of hsa_circ_0034642. For the part of functional assays, gain/loss-of-function assays were conducted to measure cell growth, apoptosis, and metastatic properties affected by hsa_circ_0034642. Furthermore, the mechanisms of hsa_circ_0034642 were investigated by dual-luciferase reporter assays. As the results demonstrated, hsa_circ_0034642 was boosted in glioma tissues and cells. Overexpression of hsa_circ_0034642 was associated with clinical severity and poor prognosis. What is more, elevated hsa_circ_0034642 strikingly facilitated cell proliferation, migratory and invasive capacities, and decreased apoptotic cells. Mechanistically, hsa_circ_0034642 sponges miR-1205. miR-1205 regulates BATF3 level through targeting its 3'-untranslated region. In summary, hsa_circ_0034642 might play a key role in this malignancy.

Long-term outcome of microvascular decompression for hemifacial spasm.

AIM: To investigate the long term outcomes of microvascular decompression (MVD) for hemifacial spasm (HFS) and to identify any prognostic factors. METHODS: A retrospective analysis of 189 consecutive patients with typical HFS who underwent MVD. Multiple logistic regression analysis of variables at various time points including at least immediate time point and one at no less than six years was performed. RESULTS: Short-term follow-up showed a cure rate of 91%, including 51 cases of delayed resolution (27%). At two years or more information was available in 148 (out of 189) cases of patients. 101 cases (68% - of 148 cases) had complete recovery, 28 cases (19%) achieved a partial though worthwhile recovery, so that the effective rate of symptoms relief at six years was 87%. Complications were found (66/189, 34.92%) and cured within the follow-up period (cure rate of 100%). In both the univariate and multivariate analyses, the postoperative findings of clinical outcomes showed that preoperative illness duration, compressive pattern, the intraoperative indentation of the root exit zone (REZ) of the facial nerve and intraoperative AMR disappearance were negative predictors and age considered to be positive, which significantly predicted the clinical outcome of patients following MVD. CONCLUSIONS: MVD may be a safe and effective strategy for HFS patients in view of relatively higher cure rates and lower complication risks within follow-up. Besides, patients' age, duration of disease, intraoperative indentation of the REZ of the facial nerve, and disappearance of AMR were the major influential variables may be useful for the prediction of prognosis in the patients underwent MVD.

A game-theoretical approach to multimedia social networks security.

The contents access and sharing in multimedia social networks (MSNs) mainly rely on access control models and mechanisms. Simple adoptions of security policies in the traditional access control model cannot effectively establish a trust relationship among parties. This paper proposed a novel two-party trust architecture (TPTA) to apply in a generic MSN scenario. According to the architecture, security policies are adopted through game-theoretic analyses and decisions. Based on formalized utilities of security policies and security rules, the choice of security policies in content access is described as a game between the content provider and the content requester. By the game method for the combination of security policies utility and its influences on each party's benefits, the Nash equilibrium is achieved, that is, an optimal and stable combination of security policies, to establish and enhance trust among stakeholders.