Morphological characteristics and phylogenetic analyses revealed four new wood inhabiting fungi (Agaricomycetes, Basidiomycota) in Xizang Autonomous Region, China.

Four new fungi from Xizang in southwest China, Caloceraramaria, Ceraceomycesrhizomorphus, Leptosporomyceslinzhiensis, and Ramariaxizangensis are described and illustrated based on the morphological and molecular evidence. Caloceraramaria is characterized by the ramal and bright orange basidiomata, a monomitic hyphal system with simple septa generative hyphae, usually 4-septate basidiospores; Ceraceomycesrhizomorphus is characterized by the cream to yellowish basidiomata with rhizomorphs, cylindrical basidiospores; Leptosporomyceslinzhiensis is characterized by white with pink basidiomata, cylindrical to oblong ellipsoid basidiospores; Ramariaxizangensis is characterized by flesh pink basidiomata, branched dichotomously in 4-5 ranks, a monomitic hyphal system with clamped generative hyphae, ellipsoid to cylindrical and densely warted basidiospores.

Systemic immune-inflammation index, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio in patients with type 2 diabetes at different stages of diabetic retinopathy.

AIM: To investigate systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) levels in patients with type 2 diabetes at different stages of diabetic retinopathy (DR). METHODS: This retrospective study included 141 patients with type 2 diabetes mellitus (DM): 45 without diabetic retinopathy (NDR), 47 with non-proliferative diabetic retinopathy (NPDR), and 49 with proliferative diabetic retinopathy (PDR). Complete blood counts were obtained, and NLR, PLR, and SII were calculated. The study analysed the ability of inflammatory markers to predict DR using receiver operating characteristic (ROC) curves. The relationships between DR stages and SII, PLR, and NLP were assessed using multivariate logistic regression. RESULTS: The average NLR, PLR, and SII were higher in the PDR group than in the NPDR group (P=0.011, 0.043, 0.009, respectively); higher in the NPDR group than in the NDR group (P<0.001 for all); and higher in the PDR group than in the NDR group (P<0.001 for all). In the ROC curve analysis, the NLR, PLR, and SII were significant predictors of DR (P<0.001 for all). The highest area under the curve (AUC) was for the PLR (0.929 for PLR, 0.925 for SII, and 0.821 for NLR). Multivariate regression analysis indicated that NLR, PLR, and SII were statistically significantly positive and independent predictors for the DR stages in patients with DM [odds ratio (OR)=1.122, 95% confidence interval (CI): 0.200-2.043, P<0.05; OR=0.038, 95%CI: 0.018-0.058, P<0.05; OR=0.007, 95%CI: 0.001-0.01, P<0.05, respectively). CONCLUSION: The NLR, PLR, and SII may be used as predictors of DR.

Morphological and molecular identification for four new wood-inhabiting species of Trechispora (Basidiomycota) from China.

Four new wood-inhabiting fungi, Trechisporaalbofarinosa, T.bisterigmata, T.pileata and T.wenshanensisspp. nov., are proposed based on a combination of morphological features and molecular evidence. Trechisporaalbofarinosa is characterized by the farinose basidiomata with flocculence hymenial surface, a monomitic hyphal system with clamped generative hyphae, and ellipsoid, warted basidiospores. Trechisporabisterigmata is characterized by the membranous basidiomata with odontioid hymenial surface, rhizomorphic sterile margin, barrelled basidia and subglobose to broad ellipsoid, smooth basidiospores. Trechisporapileata is characterized by the laterally contracted base, solitary or imbricate basidiomata, fan shaped pileus, radially striate-covered surface with appressed scales, odontioid hymenophore surface, and subglobose to broad ellipsoid, thin-walled, smooth basidiospores. Trechisporawenshanensis is characterized by a cottony basidiomata with a smooth hymenial surface, and ellipsoid, thin-walled, warted basidiospores. Sequences of ITS and LSU marker of the studied samples were generated, and phylogenetic analyses were performed with the maximum likelihood, maximum parsimony, and Bayesian inference methods. The phylogenetic tree inferred from the ITS+nLSU sequences highlighted that four new species were grouped into the genus Trechispora.

Two-dimensional speckle-tracking echocardiography in left ventricular systolic function in patients with systemic lupus erythematosus.

OBJECTIVES: To investigate whether two-dimensional speckle-tracking echocardiography (2DSTE) can be considered a criterion for early left ventricular (LV) systolic impairment in patients with systemic lupus erythematosus (SLE) and to further explore the association with each other. METHODS: We included 38 patients with SLE and assessed the degree of disease activity according to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) 2000 scoring criteria, together with 38 healthy controls who were matched by sex and age. Routine LV systolic function evaluation parameters were obtained by echocardiography as well as 2DSTE measurement of LV strain parameters to obtain global longitudinal strain (GLS) values, respectively. RESULTS: (I) On routine LV function parameters such as ejection fractions (EF) and left ventricular end-diastolic internal diameter (LVIDd), the SLE group and the control group did not reflect differences. In contrast, on the LV strain parameter obtained from 2DSTE measurements, the GLS values in all cardiac planes were lower in the SLE group than in the control group and showed statistically significant differences. (II) Correlation analysis showed that there was a correlation between SLEDAI and GLS, especially a meaningful correlation with GLS Avg and GLS A4C, with correlation coefficients of 0.35 and 0.47, respectively. CONCLUSIONS: The use of 2DSTE can detect early impaired LV systolic function in SLE patients, and GLS is progressively gaining attention as an indicator of subclinical myocardial injury and LV function in SLE patients. The correlation that exists between GLS and SLEDAI might contribute to a better assessment of cardiac involvement in SLE patients. Key Points • Cardiac involvement has become one of the major factors in the poor prognosis of SLE patients, which directly affects the mortality of SLE patients. Traditional echocardiography is difficult to detect early left ventricular function impairment, thus affecting clinicians' judgment and diagnosis. • 2DSTE can recognize subclinical myocardial injury in SLE patients at an early stage, and its derived strain parameters may be used as an indicator to evaluate myocardial involvement and reflect disease activity in SLE patients.

Qntrolling the LncRNA HULC-Tregs-PD-1 axis inhibits immune escape in the tumor microenvironment.

Background: Immune escape remains a major challenge in the treatment of malignant tumors. Here, we studied the mechanisms underlying immune escape in the tumor microenvironment and identified a potential therapeutic target. Methods: Pathological specimens from patients with liver cancer, soft tissue sarcoma, and liver metastasis of colon cancer were subjected to immunohistochemistry analysis to detect the expression of programmed death-1 (PD-1) in the tumor microenvironment (TME). Additionally, the expression of regulatory T cells (Tregs) and long non-coding RNAs (lncRNAs), such as highly upregulated in liver cancer (HULC) was evaluated by fluorescence in situ hybridization, and the relationship between HULC, Treg cells, and PD-1 was determined. The animals were divided into H22 hepatic carcinoma and S180 sarcoma groups. Each group was divided into Foxp3-/-C57BL/6J and C57BL/6J mice. Thereafter, mice were inoculated with 0.1 ml S180 sarcoma cells or 0.1 ml H22 hepatoma cells, at a concentration of 1 × 107/ml. The number of splenic CD4+CD25+Foxp3+ T cells was detected by flow cytometry, and serum interleukin-10 (IL-10) and transforming growth factor ß1 (TGF-ß1) levels were detected using a Luminex liquid suspension chip. Expression of PD-1, fork head box P3 (Foxp3), and HULC in the TME, were analyzed and the therapeutic effect of inhibiting the lncRNA HULC-Treg-PD-1 axis in malignant tumors was determined. Results: High expression of lncRNA HULC promotes the proliferation of Treg cells and increases PD-1 expression in the tumor microenvironment. The HULC-Treg-PD-1 axis plays an immunosuppressive role and promotes the proliferation of malignant tumors. Knocking out the Foxp3 gene can affect the HULC-Treg-PD-1 axis and reduce PD-1, IL-10, and TGF-ß1 expression to control the growth of malignant tumors. Conclusion: The lncRNA HULC-Treg-PD-1 axis promotes the growth of malignant tumors. This axis could be modulated to reduce PD-1, IL-10, and TGF-ß1 expression and the subsequent immune escape. The inhibition of immune escape in the tumor microenvironment can be achieved by controlling the LncRNA HULC-Treg-PD-1 axis.

Species diversity and taxonomy of Vararia (Russulales, Basidiomycota) with descriptions of six species from Southwestern China.

Vararia is a species-rich genus in the family Peniophoraceae and has been shown to be polyphyletic. In this study, sequences of ITS and LSU rRNA markers of the studied samples were generated and phylogenetic analyses were performed with the maximum likelihood, maximum parsimony, and Bayesian inference methods. Seventeen lineages including six new species from China, i.e., V.fissurata, V.lincangensis, V.punctata, V.isabellina, V.sinensis, and V.yaoshanensis were recognized, in which V.fissurata is characterized by the brittle basidiomata with pruinose and cracking hymenophore having white to olivaceous buff hymenial surface, the clamped generative hyphae, presence of the two types gloeocystidia; V.lincangensis is characterized by the simple-septa generative hyphae, and thick-walled skeletal hyphae, and ellipsoid basidiospores; V.punctata is delimited by its thin to slightly thick-walled generative hyphae, and thick-walled skeletal hyphae, present thick-walled, clavate to cylindrical gloeocystidia; V.isabellina is characterized by having the cream to isabelline to slightly brown hymenial surface, thin to slightly thick-walled generative hyphae, and sub-fusiform to navicular basidiospores; V.sinensis is distinguishable by its white to slightly pink hymenial surface, thick-walled skeletal hyphae, and sub-fusiform to navicular basidiospores; V.yaoshanensis is characterized by cream to pinkish buff to cinnamon-buff hymenial surface, slightly thick-walled generative hyphae, the presence of two types gloeocystidia, and slightly thick-walled, ellipsoid basidiospores. Phylogram based on the ITS+nLSU rDNA gene regions included nine genera within the family Peniophoraceae as Amylostereum, Asterostroma, Baltazaria, Dichostereum, Michenera, Peniophora, Scytinostroma and Vararia, in which the six new wood-inhabiting fungi species were grouped into genus Vararia. The phylogenetic tree inferred from the combined ITS and LSU tree sequences highlighted that V.fissurata was found to be the sister to V.ellipsospora with strong supports. Additionally, V.lincangensis was clustered with V.fragilis. Furthermore, V.punctata was retrieved as a sister to V.ambigua. Moreover, V.sinensis was grouped with five taxa as V.breviphysa, V.pirispora, V.fusispora, V.abortiphysa and V.insolita. The new species V.isabellina formed a monophyletic lineage, in which it was then grouped closely with V.daweishanensis, and V.gracilispora. In addition, V.yaoshanensis was found to be the sister to V.gallica with strong supports. The present results increased the knowledge of Vararia species diversity and taxonomy of corticioid fungi in China. An identification key to 17 species of Vararia in China is provided.

Relationship between lymphocytes and idiopathic macular hole.

BACKGROUND: An idiopathic macular hole (IMH) is a full-thickness anatomic defect extending from the internal limiting membrane to the photoreceptor layer of the macula without any known cause. Recently, clinical laboratory markers of systemic inflammatory status derived from complete blood counts have been evaluated in ocular diseases. This study aimed to explore whether they could predict the development and progression of IMHs. METHODS: A retrospective review of 36 patients with IMH and 36 sex-and-age-matched patients with cataracts was conducted. We collected complete blood counts of all participating individuals and calculated systemic immunoinflammatory indicators. The maximum base diameter of the IMH (BD), minimum diameter of the IMH (MIN), height of the IMH (H), area of the intraretinal cyst (IRC), and curve lengths of the detached photoreceptor arms were measured on optical coherence tomography (OCT) images. We used these values to calculate the macular hole index (MHI), tractional hole index (THI), diameter hole index (DHI), hole form factor (HFF), and macular hole closure index (MHCI). We performed a receiver operating characteristic (ROC) curve analysis of 30 patients with IMH who were followed up 1 month after surgery. RESULTS: Lymphocyte counts were significantly higher in the IMH group. No other significant differences were observed between the IMH and control groups. Lymphocyte counts in the IMH group were significantly negatively correlated with MIN and BD and were significantly positively correlated with MHI, THI, and MHCI. However, lymphocyte counts were not significantly correlated with H, IRC, DHI, and HFF. In the ROC analysis, BD, MIN, MHI, THI, and MHCI were significant predictors of anatomical outcomes. According to the cut-off points of the ROC analysis, lymphocyte counts were compared between the above-cut-off and below-cut-off groups. Lymphocyte counts were significantly higher in the MIN ≤ 499.61 µm, MHI ≥ 0.47, THI ≥ 1.2, and MHCI ≥ 0.81 groups. There were no significant differences between the above-cut-off and below-cut-off BD groups. CONCLUSIONS: Although inflammation may not be an initiating factor, it may be involved in IMH formation. Lymphocytes may play a relatively important role in tissue repair during the developmental and postoperative recovery phases of IMH.

First Report of Panax notoginseng Leaf spot Disease Caused by Boeremia exigua in China.

Panax notoginseng a perennial herb native to China, is widely grown in the Yunnan Province. (Yang et al. 2022). From July to August 2022, a new leaf spot disease was observed on fully expanded leaves of P. notoginseng from a planting base in the Xundian, Yunnan Province, China. Approximately 250 ha. of P. notoginseng is the cultivated area, and the incidence of leaf spot disease was around 10-15%. Round spots appeared on the infected leaves and as the disease progressed these leaves fell off the plant. A total 21 symptomatic leaves were randomly collected from the planting base to isolate the pathogens and further study in the laboratory. The surface of infected leaves were sanitized with 0.5% sodium hypochlorite for 2 min. and 75% alcohol for 1 min., and then rinsed thrice with sterile water. Once drying, the samples were placed on potato dextrose agar (PDA), plates and incubated at 25 °C for 5 days. The fungus was isolated from the symptomatic tissue, but only three isolates were preserved for further identification. Pure cultures of the representative strain Zhaochanglin 118 were obtained using the singlespore method, and the colonies obtained were dark-green to dark-black in appearance. The pycnidia were dark brown, solitary, or congregated with an inconspicuous neck. The conidia were colorless, ellipsoidal, and measured between 4.5 to 7 × 2 to 3 µm (n = 30). These morphological characteristics were similar to those described for Boeremia exigua (Valenzuela-Lopezi et al. 2018). The genomic DNA of the isolate was extracted using the DN14 cetyltrimethylammonium bromide rapid plant genome extraction kit. The internal transcribed spacer (ITS), RNA polymerase second largest subunit (RPB2) and translation elongation factor 1-alpha (TEF1) genes were amplified via polymerase chain reaction using the primers ITS1/ITS4 (White et al. 1990), Af/Cf (Matheny et al. 2002), and EF1-983F/EF1-2218R (Chen et al. 2015), respectively. All sequences were deposited in GenBank (OQ996531 for ITS; OR291158 for RPB2 and OR291159 for TEF1). A BLASTN homology search using the ITS nucleotide sequence indicated that this has 99.6% identity with the sequence MH859059, named B. exigua from CBS culture collection (517/519 bp); the RPB2 sequence has 97.5% identity with sequence GU371780, named B. exigua from CBS culture collection (704/722 bp); and the TEF1 sequence has 98.4% identity with sequence GU349080, named B. exigua from CBS culture collection (871/885 bp). To test Koch's postulates, a pathogenicity test was carried out on the leaves of six fully expanded P. notoginseng plants in the Xundian planting base. Conidial suspensions were prepared for one isolates at a concentration of 106 spores per milliliter. Three leaflets on different plants were applied with 20µl spore suspension and the other three leaflets were drop of 20 µl sterile distilled water. The whole experiment was repeated three times. The P. notoginseng plants were incubated under sterile conditions at 25°C for 7 days. Inoculated leaves showed the characteristic brown round spots, while control leaves were asymptomatic so, Koch's postulates were fulfilled by re-isolating the pathogen from symptomatic tissue, which was subsequently confirmed as B. exigua through morphological and molecular analyses. Koch's postulates were fulfilled. To our knowledge, this is the first report of B. exigua causing leaf spot disease in P. notoginseng in China, which lays a foundation for further study and developing disease control methods.

Four New Fungal Species in Forest Ecological System from Southwestern China.

Four new wood-inhabiting fungi were found in Southwestern China within the genera Phanerochaete, Phlebiopsis, Asterostroma, and Vararia of the families Phanerochaetaceae and Peniophoraceae, belonging to the orders Polyporales and Russulales individually. Combined with their morphological characteristics and molecular biological evidence, the present study describes them as new fungal taxa. Asterostroma yunnanense is characterized by the resupinate, membranaceous to pellicular basidiomata with a cream to salmon-buff hymenial surface, hyphal system dimitic bearing simple-septa, thin- to thick-walled, yellowish brown asterosetae with acute tips, and thin-walled, echinulate, amyloid, globose basidiospores. Phanerochaete tongbiguanensis is characterized by the resupinate basidiomata with a white to cream hymenial surface, a monomitic hyphal system with simple-septa generative hyphae, the presence of subclavate cystidia covered with a lot of crystals, and oblong ellipsoid basidiospores (6-9 × 3-4.5 µm). Phlebiopsis fissurata is characterized by the membranaceous, tuberculate basidiomata with a buff to slightly brown hymenial surface, a monomitic hyphal system with simple-septa, conical cystidia, and broadly ellipsoid. Vararia yingjiangensis is characterized by a corky basidiomata with a pinkish buff to cinnamon-buff hymenial surface, cracking, yellowish dichohyphae with slightly curved tips, subulate gloeocystidia, and thick-walled, ellipsoid basidiospores (6.5-11.5 × 5-7 µm). The phylogenetic analyses of ITS + nLSU revealed that the two new species were nested into the genera Phanerochaete and Phlebiopsis within the family Phanerochaetaceae (Polyporales), in which Phanerochaete tongbiguanensis was sister to P. daliensis; Phlebiopsis fissurata was grouped with P. lamprocystidiata. Two new species were clustered into the genera Asterostroma and Vararia within the family Peniophoraceae (Russulales), in which Asterostroma yunnanense was sister to A. cervicolor; Vararia yingjiangensis formed a single branch.

Phylogenetic and Taxonomic Analyses of Five New Wood-Inhabiting Fungi of Botryobasidium, Coltricia and Coltriciella (Basidiomycota) from China.

In this present study, five new wood-inhabiting fungal taxa, Botryobasidium gossypirubiginosum, Botryobasidium incanum, Botryobasidium yunnanense, Coltricia zixishanensis, and Coltriciella yunnanensis are proposed. Botryobasidium gossypirubiginosum is distinguished by its slightly rubiginous hymenial surface, monomitic hyphal system, which branches at right angles, and subglobose, smooth basidiospores (14-17.5 × 13-15.5 µm); B. incanum is characterized by its white to incanus basidiomata having a hypochnoid hymenial surface, and ellipsoid, smooth basidiospores (6.5-8.5 × 3.5-5 µm); B. yunnanense is characterized by its buff to slightly yellowish hymenial surface, monomitic hyphal system, and broadly ellipsoid to globose, smooth, thick-walled basidiospores (11.5-14.5 × 9.5-10.5 µm); Coltricia zixishanensis differs in its rust brown pileal surface, and ellipsoid, thick-walled basidiospores (5-6.5 × 4-4.5 µm). Coltriciella yunnanensis is distinguished by its tiny pilei, short stipe, and navicular, verrucose basidiospores (10.5-12.5 × 6-7 µm). Sequences of ITS and nLSU genes were used for phylogenetic analyses using the maximum likelihood, maximum parsimony, and Bayesian inference methods. The phylogenetic results inferred from ITS sequences revealed that B. gossypirubiginosum was closely related to B. robustius; the species B. incanum was grouped with B. vagum; B. yunnanense was related to B. indicum. The species C. zixishanensis was grouped with C. confluens and C. perennis. ITS sequences revealed that C. zixishanensis was grouped into the genus Coltriciella, in which it was grouped with Co. globosa and Co. pseudodependens.

New species of Tropicoporus (Basidiomycota, Hymenochaetales, Hymenochaetaceae) from India, with a key to Afro-Asian Tropicoporus species.

The Inonotuslinteus complex, predominantly reported from East Asia, Mesoamerica and Caribbean countries, was circumscribed into Tropicoporus as one of the new genera, based on morphological and phylogenetic data. The present paper describes four new species of Tropicoporus from India. Morphological characteristics and phylogenetic analyses, based on ITS and nLSU data, delimited the new species, which are named T.cleistanthicola, T.indicus, T.pseudoindicus and T.tamilnaduensis. The pairwise homoplasy index (PHI) test was done to confirm the distinctive nature of the new species. The traits of Indian species remain distinct from one another, except for the pileate basidiome with the mono-dimitic hyphal system, cystidioles and broadly ellipsoid basidiospores. Descriptions, illustrations, PHI test results and a phylogenetic tree to show the position of the new species are provided. In addition, an identification key to Tropicoporus in Asia and an African species is given.

Phylogenetic and Taxonomic Analyses Reveal Three New Wood-Inhabiting Fungi (Polyporales, Basidiomycota) in China.

Three new wood-inhabiting fungal species, Cerioporus yunnanensis, Perenniporiopsis sinensis, and Sarcoporia yunnanensis, are proposed based on a combination of the morphological features and molecular evidence. Cerioporus yunnanensis is characterized by the pileate basidiomata having a fawn brown to black pileal surface, a dimitic hyphal system with clamped generative hyphae, and the presence of the fusoid cystidioles and cylindrical basidiospores (9-12.5 × 3.5-5 µm). Perenniporiopsis sinensis is distinct from the osseous pileus with verrucose, an orange-yellow to dark reddish-brown pileal surface with a cream margin, a trimitic hyphal system with clamped generative hyphae, and the presence of the fusiform cystidioles and ellipsoid basidiospores (9-11 × 5.5-6.5 µm). Sarcoporia yunnanensis is typical of the pileate basidiomata with a salmon to reddish-brown pileal surface, a monomitic hyphal system with clamped generative hyphae, and the presence of the ellipsoid basidiospores (4-5.5 × 2.5-4 µm). Sequences of ITS + nLSU + mt-SSU + TEF1 + RPB1 + RPB2 genes were used for the phylogenetic analyses using maximum likelihood, maximum parsimony, and Bayesian inference methods. The multiple genes with six loci analysis showed that the three new species nested within the order Polyporales, in which C. yunnanensis and P. sinensis nested into the family Polyporaceae, and S. yunnanensis grouped into the family Sarcoporiaceae.

First Report of Brown Rot Caused by Porogramme epimiltina on Gastrodia elata in China.

Gastrodia elata Blume is a valuable medicinal plant in China with great significance in medicine (Li et al. 2023). From 2022 to 2023, G. elata tuber rot occurred in about 50 households in the main cultivation areas of G. elata (27°39' N, 104°16' E) in Yiliang County, Zhaotong, Yunnan Province, southwest China. The planting area of G. elata was 776 ha, and the incidence rate was 10%. Symptoms present as light brown lesions on the surface of the tuber, sunken, soft and foul-smelling. Infected G. elata tubers were randomly collected from each household, packed into transparent plastic bags, and strains were isolated in the laboratory as follows. The tubers of 15 infected G. elata tubers were surface-sterilized with 0.5% NaOCl for 2 min, rinsed five times with sterile water, and dried. Symptomatic tissues from the margin between necrotic and healthy tissues were cut into 5 × 5 mm pieces, placed onto potato dextrose agar (PDA), and incubated at 28 ºC in the dark for 3 days. Hyphal tips of fungi growing from the samples were transferred onto new PDA plates and incubated until they produced conidia. Two fungal strains (Charliezhao 425 and 433) with the same morphological characteristics were obtained from the samples. Colonies were whitish and grew rapidly, irregularly turning pale orange at the edge or center of the mycelium pad on a two-week-old petri dish, and finally dark red，spore oval to spherical, 2.7 to 5.3 × 2.3 to 3.5 µm (n=50). The morphological characteristics of the isolates resembled Porogramme epimiltina (Mao et al. 2023; Kubayashi et al. 2001). Genomic DNA of two representative isolates (Charliezhao 425 and 433) was extracted using the DN14 cetyltrimethylammonium bromide rapid plant genome extraction kit (Aidlab Biotechnologies Co., Ltd, Beijing). The ITS and TEF1 genes were amplified by polymerase chain reaction using the primers ITS1/ITS4 (White et al. 1990) and EF1-983F/EF1-2218R (Rehner et al, 2005), respectively. All sequences were deposited in GenBank (accession no. OR905803, OR905804 for ITS, OR939812, OR939813 for TEF1). A BLASTN homology search with the ITS nucleotide sequences showed that they had 98.99 to 99.15% identity with P. epimiltina isolate OP997539 (588/594 bp) and isolate OP997539 (584/589 bp), respectively; and the TEF1 sequences had 95.41 to 95.59% % identity to isolates OP556566 (540/565 bp) and isolate OP556566 (542/567 bp), respectively. To complete Koch's hypothesis, the surfaces of 5 mature and healthy G. elata tubers were disinfected with 1% NaClO solution for 1 minute, rinsed with sterile water 5 times, and dried at 25 ℃ for 30 minutes. Conidial suspensions (106 spores/ml) were collected from two isolates (Charliezhao 425 and 433) and sprayed on G. elata tuber, and the control treated with distilled water. All G. elata tubers were incubated at 25℃ with 80% relative humidity. The experiment had three replicates. After 7 days of culture, there were obvious rotten and smelly on the inoculated tubers. No symptoms were observed in the control groups. The pathogen was re-isolated from all inoculated birch tubers and confirmed as P. epimiltina by morphological and molecular analysis, which fulfilled Koch's hypothesis. Gastrodia elata is a valuable and extensively used herbal Traditional Chinese Medicine with a wide range of clinical applications. As far as we know, this is the first report of P. epimiltina causing brown rot of G. elata in China.

Molecular Systematics and Taxonomic Analyses of Three New Wood-Inhabiting Fungi of Hyphoderma (Hyphodermataceae, Basidiomycota).

In this present study, three new wood-inhabiting fungal taxa, Hyphoderma niveomarginatum, H. sordidum and H. weishanense, are proposed. Hyphoderma niveomarginatum is characterized by the ceraceous basidiomata having a smooth, cracking hymenial surface and the presence of the moniliform cystidia and ellipsoid basidiospores (7-9 × 3.5-5 µm). Hyphoderma sordidum is characterized by its resupinate basidiomata with a smooth hymenial surface with the fimbriate margin, the presence of the tubular cystidia and ellipsoid basidiospores (3-4.5 × 2-3 µm). Hyphoderma weishanense differs in its membranous basidiomata with a slightly buff to buff hymenial surface and the presence of broadly ellipsoid basidiospores (4.5-8.5 × 4-7 µm). Sequences of ITS+nLSU+mt-SSU+RPB1+RPB2 genes were used for the phylogenetic analyses using three methods. The ITS+nLSU+mt-SSU+RPB1+RPB2 analysis of the genus Hyphoderma indicated that the 3 new species of Hyphoderma were nested into genus Hyphoderma, in which H. niveomarginatum formed a single group and then grouped with H. membranaceum and H. sinense; H. sordidum was a sister to H. nudicephalum; and H. weishanense closely grouped with H. crystallinum.

First Report of Mucor Rot Caused by Mucor circinelloides on Gastrodia elata in China.

Gastrodia elata Blume is used as a traditional Chinese herbal medicine and is widely planted throughout China (Zhu et al. 2019). From September to October 2022, G. elata tuberous rot occurred in 47 households in Yiliang County, Zhaotong City (27°39' N, 104°16' E), Yunnan Province, China, within a major G. elata production area covering 776 ha, with an incidence rate of 10 to 15%. Symptoms manifested as black and brown lesions on the tuber surface, which were concave, soft, foul-smelling, and surrounded by grayish-brown tissue . Three infected G. elata were randomly collected from each household, placed in transparent plastic bags, and pathogen isolation was conducted in a laboratory. Infected G. elata tubers were surface-sterilized with 0.5% NaOCl for 2 min, rinsed five times with sterile water, and dried. Symptomatic tissues from the margin between necrotic and healthy tissues were cut into 5 × 5 mm pieces, placed onto potato dextrose agar (PDA), and incubated at 28 ºC in the dark for 3 days. Hyphal tips of fungi growing from the samples were transferred onto new PDA plates and incubated until they produced conidia. Five fungal isolates (Charliezhao 417, 418, 419, 420, and 421) with the same morphological characteristics were obtained from the samples. Colonies tended to be yellow or light grey, and produced sporangiospores that were sub-globose, ellipsoid, or irregular, measuring 4.0 to 9.7 × 2.5 to 4.6 µm (n = 50). The morphological characteristics of the isolates resembled Mucor circinelloides (Wagner et al. 2020). Genomic DNA of two representative isolates (Charliezhao 417 and 418) was extracted using the DN14 cetyltrimethylammonium bromide rapid plant genome extraction kit (Aidlab Biotechnologies Co., Ltd, Beijing). The ITS and RPB1 genes were amplified by polymerase chain reaction using the primers ITS1/ITS4 (White et al. 1990) and Af/Cf (Matheny et al. 2002), respectively. All sequences were deposited in GenBank (accession no. OQ612709, OR028949 for ITS, OQ621439, OR033135 for RPB1). A BLASTN homology search with the ITS nucleotide sequences showed that they had 99.5 to 99.2% identity with M. circinelloides isolate KR056083 (603/606 bp) and isolate KJ588204 (617/622 bp), respectively; and the RPB1 sequences had 99.89% to 99.75% identity to isolates KJ588206 (874/875 bp) and isolate KJ588206 (803/805 bp), respectively. To complete Koch's postulates, five mature, healthy G. elata tubers were surface disinfected with 1% NaClO solution for 1 min, rinsed with sterile water, and dried at 25℃ for 30 min. A conidial suspension (106 spores/ml) collected from two isolates (Charliezhao 417 and 418) was sprayed onto G. elata tubers, and the control treated with distilled water. All G. elata tubers were incubated at 25 ºC with 80% relative humidity. The experiment had five replicates. After 7 days of incubation, there were obvious brown spots on inoculated tubers; no symptoms were observed on the controls. The pathogen was re-isolated from all inoculated G. elata tubers and confirmed as M. circinelloides by morphological and molecular analyses, completing Koch's postulates. This is the first report of M. circinelloides causing G. elata mucor rot in China. The tubers of G. elate are often employed in the treatment of headaches, convulsions and neurodegenerative disorders (Manavalan et al. 2012). Thus, the declining yield of G. elate due to persistent obstacles related to continuous cropping and diseases poses a potential threat to the development of the G. elate industry.

Molecular phylogeny and morphology reveal a new wood-rotting fungal species, Sistotrema yunnanense sp. nov. from the Yunnan-Guizhou Plateau.

Wood-rotting fungi are important components of woody plant ecosystems and play an active role in the decomposition and turnover of nutrients from wood, and are among the major groups of Basidiomycota. In this study, a new species of wood-rotting fungus, Sistotrema yunnanense, was proposed based on morphological characteristics and molecular evidence. It is characterized by resupinate basidiomata, a monomitic hyphal system having generative hyphae with clamp connections, suburniform to urniform basidia, and short-cylindrical to oblong ellipsoid basidiospores (4.5-6.5 × 3-4 µm). Phylogenetic analyses performed using the large subunit nuc rDNA indicated that S. yunnanense was nested within the genus Sistotrema s.l. of the family Hydnaceae, within the order Cantharellales.

First Report of Panax notoginseng Wilt Disease Caused by Fusarium graminearum in China.

Panax notoginseng is one of the important economic crops under the forest, which is widely planted in Yunnan Province, China. In August of 2022, a survey in Xundian county (25º26' N, 12 103°7' E), was accomplished to verify the occurrence of wilt disease in P. notoginseng and understand its aetiology. The site is an underforest of organic P. notoginseng, covering an area of over 40 ha. Disease symptoms included severe stunting, leaf chlorosis, red or yellow stalks, and rotting roots. The entire plant gradually wilted and died with disease progression (Fig. 1). To identify the causal agent, we collected more than 30 wilted P. notoginseng plants and got the plant tissues from the symptomatic leaves, stalks, and roots. The tissues surface sterilised with 0.5% sodium hypochlorite for 2 min, followed by 75% alcohol for 1 min, and rinsed in sterilised water three times. Upon drying, samples were placed onto potato dextrose agar (PDA) incubated in the dark at 25°C (Bilgi et al. 2011). Isolates were then transferred to carnation leaf agar (CLA) to induce sporulation. Colonies on PDA were yellow, orange to red, with abundant fluffy aerial mycelia with a dark red pigment on the undersides; Colonies on CLA were orange to yellow (Fig. 2). Fusiform macroconidia and bottle-shaped conidiogenous cells were visible under a microscope. Microconidia were not observed. Macroconidia were measured as 18.5-40.5 µm × 3-4.7 µm (n = 60) (Fig. 3), and possessed 2 to 6 septa. These are similar to previously reported morphological characteristics of Fusarium graminearum (Shikur et al. 2018; Martinez et al. 2019). Cetyltrimethylammonium bromide rapid plant genome extraction kit-DN14 was used to obtain genomic DNA from two representative isolate, the ITS, TEF1 and RPB2 gene were amplified by Polymerase Chain Reaction using primers ITS5/ITS4 (White et al, 1990), EF1-983F/EF1-2218R (Rehner et al, 2005), bRPB2-6F/bRPB2-7.1R (Matheny et al, 2002), respectively. BLAST homology search for nucleotide sequences revealed > 99% similarity to F. graminearum ITS (550bp; MG274308, KU847854), TEF1 (1000bp; MH572248, MH572252) and RPB2 (1000bp; KT855203, KT855206) sequences. All sequences from this study were deposited in GenBank (OP617343 and OP617344 for ITS; OP930951 and OP930952 for TEF1; OP930953 and OP930954 for RPB2). In the phylogenetic tree, the isolates (SWFU 0000116, SWFU 0000117) clustered with the representative strains of F. graminearum. The morphology and multi-gene phylogenetic analysis indicated that the new isolate is F. graminearum. Koch's postulates were used to confirm that the symptoms in wilted P. notoginseng were attributable to F. graminearum. First, healthy leaves were gently wounded with a needle and sprayed with spore suspension (1.0 × 106 spores mL-1) in a hand sprayer (Martinez et al, 2019). All P. notoginseng plants were then replanted in pots with a diameter of 20 cm (1 plants/pot) filled with mixture of sterilised soil, and incubated at 25-27°C. The blank control comprised sterile cotton soaked in sterile water and inactivated mycelia sprayed on the leaves. After 7d of incubation, all inoculated leaves and stalks developed necrosis and developed pale red mycelia, while control plants remained symptomless (Fig. 4-5). The pathogen was successfully isolated from these inoculated plants and identified as F. graminearum. Koch's postulates were implemented. To the best of our knowledge, this is the first report from China with evidence of F. graminearum infecting P. notoginseng.

Five new species of Schizoporaceae (Basidiomycota, Hymenochaetales) from East Asia.

Five new wood-inhabiting fungi, Lyomycesalbopulverulentus, L.yunnanensis, Xylodondaweishanensis, X.fissuratus, and X.puerensisspp. nov., are proposed based on a combination of morphological features and molecular evidence. Lyomycesalbopulverulentus is characterized by brittle basidiomata, pruinose hymenophore with a white hymenial surface, a monomitic hyphal system with clamped generative hyphae, and ellipsoid basidiospores. Lyomycesyunnanensis is characterized by a grandinioid hymenial surface, the presence of capitate cystidia, and ellipsoid basidiospores. Xylodondaweishanensis is characterized by an odontioid hymenial surface, a monomitic hyphal system with clamped generative hyphae, and broad ellipsoid-to-subglobose basidiospores. Xylodonfissuratus is characterized by a cracking basidiomata with a grandinioid hymenial surface, and ellipsoid basidiospores. Xylodonpuerensis is characterized by a poroid hymenophore with an angular or slightly daedaleoid configuration, and ellipsoid-to-broad-ellipsoid basidiospores. Sequences of ITS and nLSU rRNA markers of the studied samples were generated and phylogenetic analyses were performed with the maximum likelihood, maximum parsimony, and Bayesian inference methods. The phylogram based on the ITS+nLSU rDNA gene regions (Fig. 1) included six genera within the families Chaetoporellaceae, Hyphodontiaceae, Hymenochaetaceae, and Schizoporaceae (Hymenochaetales)-Fasciodontia, Hastodontia, Hyphodontia, Kneifiella, Lyomyces, and Xylodon-in which the five new species were grouped into genera Lyomyces and Xylodon. The phylogenetic tree inferred from the ITS sequences highlighted that Lyomycesalbopulverulentus formed a monophyletic lineage and was then grouped closely with L.bambusinus, L.orientalis, and L.sambuci; additionally, L.yunnanensis was sister to L.niveus with strong supports. The topology, based on the ITS sequences, revealed that Xylodondaweishanensis was retrieved as a sister to X.hyphodontinus; X.fissuratus was grouped with the four taxa X.montanus, X.subclavatus, X.wenshanensis, and X.xinpingensis; and X.puerensis was clustered with X.flaviporus, X.ovisporus, X.subflaviporus, X.subtropicus, and X.taiwanianus.

Sinisan ameliorates colonic injury induced by water immersion restraint stress by enhancing intestinal barrier function and the gut microbiota structure.

CONTEXT: Sinisan (SNS) has been used to treat psychosomatic diseases of the digestive system. But little is known about how SNS affects water immersion restraint stress (WIRS). OBJECTIVE: To study the effects of SNS on colonic tissue injury in the WIRS model. MATERIALS AND METHODS: Forty-eight Kunming (KM) mice were randomized into 6 groups (n = 8): The control and WIRS groups receiving deionized water; the SNS low-dose (SL, 3.12 g/kg/d), SNS middle-dose (SM, 6.24 g/kg/d), SNS high-dose (SH, 12.48 g/kg/d), and diazepam (DZ, 5 mg/kg/d) groups; each with two daily administrations for 5 consecutive days. The 5 treatment groups were subjected to WIRS for 24 h on day 6. The effects of SNS on colon tissue injury caused by WIRS were assessed by changes in colon histology, inflammatory cytokines, brain-gut peptides, and tight junction (TJ) proteins levels. 16S rRNA gene sequencing was used to detect the regulation of the gut microbiota. RESULTS: SNS pretreatment significantly reduced TNF-α (0.75- to 0.81-fold), IL-6 (0.77-fold), and IFN-γ (0.69-fold) levels; and increased TJ proteins levels, such as ZO-1 (4.06- to 5.27-fold), claudin-1 (3.33- to 5.14-fold), and occludin (6.46- to 11.82-fold). However, there was no significant difference between the levels of substance P (SP) and vasoactive intestinal peptide (VIP) in the control and WIRS groups. SNS regulated the composition of gut microbiota in WIRS mice. CONCLUSION: The positive effects of SNS on WIRS could provide a theoretical basis to treat stress-related gastrointestinal disorders.

Comparison of the diversity and structure of the rhizosphere microbial community between the straight and twisted trunk types of Pinus yunnanensis.

Background: Pinus yunnanensis is a major silvicultural species in Southwest China. Currently, large areas of twisted-trunk Pinus yunnanensis stands severely restrict its productivity. Different categories of rhizosphere microbes evolve alongside plants and environments and play an important role in the growth and ecological fitness of their host plant. However, the diversity and structure of the rhizosphere microbial communities between P. yunnanensis with two different trunk types-straight and twisted-remain unclear. Methods: We collected the rhizosphere soil of 5 trees with the straight and 5 trees with the twisted trunk type in each of three sites in Yunnan province. We assessed and compared the diversity and structure of the rhizosphere microbial communities between P. yunnanensis with two different trunk types by Illumina sequencing of 16S rRNA genes and internal transcribed spacer (ITS) regions. Results: The available phosphorus in soil differed significantly between P. yunnanensis with straight and twisted trunks. Available potassium had a significant effect on fungi. Chloroflexi dominated the rhizosphere soils of the straight trunk type, while Proteobacteria was predominant in the rhizosphere soils of the twisted trunk type. Trunk types significantly explained 6.79% of the variance in bacterial communities. Conclusion: This study revealed the composition and diversity of bacterial and fungal groups in the rhizosphere soil of P. yunnanensis with straight and twisted trunk types, providing proper microbial information for different plant phenotypes.