Taxonomy and phylogeny of three species in Perichaena sensu lato (Myxomycetes = Myxogastria) from China.

After Gulielmina was erected and Ophiotheca was resurrected based on some species originally included in Perichaena (Trichiaceae, Trichiales, Myxomyxetes), some specimens from the Herbarium of Fungi of Nanjing Normal University previously identified as Perichaena species were reexamined from morphological and two-gene (nuclear 18S rDNA and elongation factor-1 alpha) phylogenetic perspectives. In this study, two new myxomycete species, Gulielmina subreticulospora and Ophiotheca dictyospora, are described. Gulielmina subreticulospora shows the following character combination: branched plasmodiocarps, single peridium with circular protrusions in the inner surface, capillitium (2.4-)2.8-3.0(-3.4) µm in diameter, spores (7.4-)8.0-8.5(-9.0) µm in diameter and sub-reticulated. Ophiotheca dictyospora shows the following character combination: sessile sporocarps to short plasmodiocarps, single peridium with a densely irregular network and protrusions in the inner surface, capillitium (2.7-)3.5-5.0(-7.1) µm in diameter, uneven, decorated with spines of uneven size, spores (7.7-)8.2-8.6(-9.4) µm in diameter including obviously complete cristate reticulation with serrated edges, with deep and clear grids. Both new taxa were compared with related species and their genetic isolation was statistically tested. Moreover, a comprehensive morphological description and a detailed figure plate are provided for Perichaena verrucifera, and its phylogenetic position is determined.

High nutrient utilization and resorption efficiency promote bamboo expansion and invasion.

Bamboos are fast-growing, aggressively-spreading, and invasive woody clonal species that often encroach upon adjacent tree plantations, forming bamboo-tree mixed plantations. However, the effects of bamboo invasion on leaf carbon (C) assimilation, and nitrogen (N) and phosphorus (P) utilization characteristics remains unclear. We selected four different stands of Pleioblastus amarus invading Chinese fir (Cunninghamia lanceolata) plantations to investigate the concentrations, stoichiometry, and allometric growth relationships of mature and withered leaves of young and old bamboos, analyzing N and P utilization and resorption patterns. The stand type, bamboo age, and their interaction affected the concentrations, stoichiometry and allometric growth patterns of leaf C, N, and P in both old and young bamboos, as well as the N and P resorption efficiency. Bamboo invasion into Chinese fir plantations decreased leaf C, N, and P concentrations, C:N and C:P ratios, N and P resorption efficiency, and allometric growth exponents among leaf C, N, and P, while it only slightly altered N:P ratios. PLS-PM analysis revealed that bamboo invasion negatively impacted leaf C, N, and P concentrations, as well as N and P utilization and resorption. The results indicate that high N and P utilization and resorption efficiency, along with the mutual sharing of C, N, and P among bamboos in interface zones, promote continuous bamboo expansion and invasion. Collectively, these findings highlight the significance of N and P utilization and resorption in bamboo expansion and invasion and provide valuable guidance for the establishment of mixed stands and the ecological management of bamboo forests.

Arcyria similaris: A new myxomycete species from China.

A new myxomycete species, Arcyria similaris, was reported herein. The specimens were found and collected in the field on dead bark from Jingangtai National Geopark in Henan Province of China. This species has distinct and unique morphological characteristics, including dark grayish olive sporothecae that fade to smoke gray with age, shallow saucer-shaped cups with marked reticulations and thick papillae on the inner surface, a netted capillitium with many bulges, uniformly marked with low, dense, and irregular reticulations, and spores (8.0-)9.3-10.1(-10.9) µm in diameter, marked with sparse small warts and grouped prominent warts. Apart from a comprehensive morphological study, partial sequences of the nuclear 18S rDNA and elongation factor-1 alpha (EF-1α) genes were also provided in this study. This new species was described and illustrated morphologically. The specimens are deposited in the Herbarium of Fungi of Nanjing Normal University (HFNNU).

The evaluation of four nano-formulations loaded-Elsinochrome A on characteristics and in vitro cytotoxicity effect.

Elsinochrome A (EA) is a naturally occurring photosensitizer with potential applications in photodynamic therapy (PDT) for various malignancies. Despite its promising therapeutic properties, the poor solubility of EA hampers its effective utilization in clinical settings. To circumvent this limitation, we engineered four distinct nano-formulations: PLGA/EA nanoparticles (NPs), CMC-PLGA/EA NPs, mPEG-PCL/EA nanomicelles (NMs), and LHP-CHOL/EA nanoliposomes (NLs), all designed to enhance the solubility of EA. A comparative evaluation of these formulations, based on metrics such as particle size, Zeta potential, drug loading efficiency, and encapsulation efficiency, identified PLGA/EA NPs and mPEG-PCL/EA NMs as the most efficacious candidates. Subsequent in vitro investigations into the drug release kinetics under varying pH conditions and the impact on cell viability and apoptosis in A549 and MCF-7 cell lines were conducted. Remarkably, the maximum drug release for PLGA/EA NPs and mPEG-PCL/EA NMs was recorded at 62.5% and 70.8% in an acidic environment (pH 5.7), respectively. Upon exposure to 460 nm light, PLGA/EA NPs induced a significant reduction in A549 cell viability to 13.8% and an apoptosis rate of 93.8%, whereas mPEG-PCL/EA NMs elicited a decrease in MCF-7 cell viability to 12.8% and an apoptosis rate of 73.0%.

Elsinochrome A induces cell apoptosis and autophagy in photodynamic therapy.

Elsinochrome A (EA) is a perylene quinone natural photosensitizer, photosensitizer under light excitation generates reactive oxygen species (ROS) to induce apoptosis, so can be used for treating tumors, that is so-called photodynamic therapy (PDT). However, the molecular mechanism, especially related to apoptosis and autophagy, is still unclear. In this study, we aimed to explore the mechanism of EA-PDT-induced B16 cells apoptosis and autophagy. The action of EA-PDT on mitochondrial permeability transition pore (MPTP), mitochondrial membrane potential (MMP) and the mitochondrial function were researched by fluorescence technique and Extracellular Flux Analyzer. Illumina sequencing, tandem mass tags Quantitative Proteomics and Western Blot studied the mechanism at the gene and protein levels. The results indicated that EA-PDT had excellent phototoxicity in vitro. EA could bind to the mitochondria. EA-PDT for 5 min caused MPTP opening, MMP decreasing and abnormal mitochondrial function with a concentration-dependent characteristic. EA-PDT resulted in an increase intracellular ROS and the number of autophagosomes. Caspase2, caspase9 and tnf were upregulated, and bcl2, prkn, atg2, atg9 and atg10 were downregulated. Our results indicated that EA-PDT induced cell apoptosis and autophagy through the mediation of ROS/Atg/Parkin. This study can provide enlightenment for exploring potential targets of drug development for the PDT of melanoma.

Distribution, assembly, and interactions of soil microorganisms in the bright coniferous forest area of China's cold temperate zone.

The bright coniferous forest area in the cold temperate zone of China is a terrestrial ecosystem primarily dominated by low mountain Larix gmelinii trees. Limited information is available regarding the assembly mechanisms and interactions of microbial communities in the soil in this region. This study employed high-throughput techniques to obtain DNA from myxomycetes, bacteria, and fungi in the soil, evaluated their diversity in conjunction with environmental factors, associated them with the assembly process, and explored the potential interaction relationships between these microorganisms. The findings of our study showed that environmental factors had a more significant influence on the α and ß diversity of bacteria compared to myxomycetes and fungi. Microbial communities were influenced by environmental selection and geographical diffusion, although environmental selection appeared to have a more significant impact than geographical diffusion. Our study suggested that different microorganisms exhibited unique evolutionary patterns and may have different assembly modes within phylogenetic groups. Myxomycetes and fungi exhibited a similar assembly process that was mainly influenced by stochastic dispersal limitation and drift. In contrast, bacteria's assembly process was primarily influenced by stochastic drift and deterministic homogeneous selection. The community of myxomycetes and fungi is greatly influenced by spatial distribution and random events, while bacteria have a relatively stable population composition in specific regions and may also be subject to environmental constraints. Finally, this study revealed that Humicolopsis cephalosporioides, a fungus that exclusively resided in cold environments, may play a critical role as a keystone species in maintaining molecular ecological networks and was considered a core member of the microbiome.

Effects of vegetation succession on soil microbial stoichiometry in Phyllstachys edulis stands following abandonment.

Moso bamboo (Phyllostachys edulis) is China's most important economic bamboo species. With a continuous decline in the value of its shoots and timber and an increase in affiliated labor and production costs, many of these stands have been abandoned, resulting in the occurrence of vegetation succession. Currently, our understanding on changes in soil microbial stoichiometric and entropic effects and associated imbalances following stand abandonment is limited. Accordingly, this study explores three timescales of Ph. edulis stand abandonment (i.e., 0, 9, and 21 years) to investigate soil-microbial carbon (C), nitrogen (N), and phosphorus (P) dynamics within a 30 cm soil profile. Results showed that (1) following abandonment, vegetation succession significantly influenced soil carbon (Csoil), nitrogen (Nsoil), and phosphorus (Psoil), microbial biomass (Cmic), nitrogen (Nmic), and phosphorus (Pmic), and Csoil:Nsoil:Psoil and Cmic:Nmic:Pmic ratios. Additionally, Csoil, Nsoil, Psoil, Cmic, Nmic, Pmic all increased significantly over time following abandonment. Moreover, Csoil:Nsoil, Cmic:Pmic, and Nmic:Pmic ratios clearly increased while Csoil:Psoil, Nsoil:Psoil, and Cmic:Nmic ratios all significantly decreased. (2) Soil microbial entropy nitrogen (qMBN) and soil microbial imbalances in Cimb:Nimb increased while soil microbial entropy carbon (qMBC), soil microbial entropy phosphorus (qMBP), and soil microbial imbalances in Cimb:Pimb and Nimb:Pimb decreased over time following abandonment. (3) Redundancy analysis (RDA) indicated that Csoil:Nsoil and Cmic:Pmic ratios were key influencing factors of microbial quotient (qMB), explaining 55.35 % and 24.39 % of variation, respectively. Following abandonment, positive or negative successional impacts on Csoil:Nsoil:Psoil, microbial C, N, P stoichiometric imbalances (Cimb:Nimb:Pimb), and Csoil:Nsoil:Psoil ratios had a positive effect on qMB. Collectively, these findings highlight the importance of Csoil:Nsoil:Psoil and Cimb:Nimb:Pimb ratios in regulating qMB induced by vegetation succession following Ph. edulis abandonment, and provide valuable information for vegetation restoration and establishment of bamboo mixed forest.

Phylogeny and Taxonomic Revision of the Family Discinaceae (Pezizales, Ascomycota).

Species of Discinaceae are common macrofungi with a worldwide distribution. Some of them are commercially consumed, while a few others are reported as poisonous. Two genera were accepted in the family: the epigeous Gyromitra with discoid, cerebriform to saddle-shaped ascomata and the hypogeous Hydnotrya with globose or tuberous ascomata. However, due to discrepancies in their ecological behaviors, a comprehensive investigation of their relationship was not thoroughly explored. In this study, phylogenies of Discinaceae were reconstructed using sequence analyses of combined and separate three gene partitions (internal transcribed spacer [ITS], large subunit ribosomal DNA [LSU], and translation elongation factor [TEF]) with a matrix containing 116 samples. As a result, the taxonomy of the family was renewed. Eight genera were recognized: two of them (Gyromitra and Hydnotrya) were retained, three (Discina, Paradiscina, and Pseudorhizina) were revived, and three (Paragyromitra, Pseudodiscina, and Pseudoverpa) were newly established. Nine new combinations were made in four genera. Two new species in Paragyromitra and Pseudodiscina and an un-named taxon of Discina were described and illustrated in detail based on the materials collected from China. Furthermore, a key to the genera of the family was also provided. IMPORTANCE Taxonomy of the fungal family Discinaceae (Pezizales, Ascomycota) was significantly renewed on the basis of sequence analyses of internal transcribed spacer (ITS), large subunit ribosomal DNA (LSU), and translation elongation factor (TEF). Eight genera were accepted, including three new genera; two new species were described; and nine new combinations were made. A key to the accepted genera of the family is provided. The aim of this study is to deepen the understanding of the phylogenetic relationships among genera of the group, as well as the associated generic concepts.

Shoot Nutrition and Flavor Variation in Two Phyllostachys Species: Does the Quality of Edible Bamboo Shoot Diaphragm and Flesh Differ?

For their quality evaluation, it is essential to determine both bamboo shoot nutrition and palatability, which will have a decisive effect on their economic value and market potential. However, differences in shoot nutrition and flavor variation among bamboo species, positions, and components have not been scientifically validated. This study assessed nutritional and flavor differences in two components (i.e., shoot flesh (BSF) and diaphragm (BSD)) of two Phyllostachys species (i.e., Phyllostachys edulis and Phyllostachys violascens) and analyzed any positional variation. Results showed that BSF protein, starch, fat, and vitamin C contents were comparatively higher. Nutrient compounds in the upper shoot segment of Ph. edulis were higher and contained less cellulose and lignin. However, both species' BSD total acid, oxalic acid, and tannin contents were comparable. BSD soluble sugar and sugar:acid ratio were higher than upper BSD total amino acid, four key amino acids (i.e., essential amino acid, bitter amino acid, umami amino acid, and sweet amino acid flavor compounds), and associated ratios were all higher than BSF while also being rich in amino acids. The content and proportion of BSF essential and bitter amino acid flavor compounds in Ph. edulis were high relative to Ph. violascens. Conversely, the content and proportion of BSD umami and sweet amino acid flavor compounds were comparable to that of Ph. edulis. Our results showed that bamboo shoot quality was affected by flavor compound differences and that interspecific and shoot components interact. This study offers a new perspective to determine the formative mechanisms involved in bamboo shoot quality while providing a basis for their different usages.

Advancements in the taxonomic study of myxomycetes (Myxogastrea) in China.

Species in the class Myxomycetes (or Myxogastrea) are essential components of biodiversity and play important ecological roles in terrestrial ecosystems, especially in forests. Studies on the taxonomy and diversity of these organisms started late in China. However, significant progress in China has been made in modern taxonomic studies on myxomycetes based on long-term species surveying and specimen collecting. The existing achievements have shown that comprehensive and continuous studies on the taxonomy and diversity of myxomycetes in China have the potential to enhance global biodiversity and improve the geographic distribution pattern of myxomycetes. Therefore, building on the current research foundation and expanding myxomycete research in a wider and more in-depth approach is imperative.

Inhibitory Effects and Mechanism of Action of Elsinochrome A on Candida albicans and Its Biofilm.

Biofilm-associated Candida albicans infections, the leading cause of invasive candidiasis, can cause high mortality rates in immunocompromised patients. Photodynamic antimicrobial chemotherapy (PACT) is a promising approach for controlling infections caused by biofilm-associated C. albicans. This study shows the effect of Elsinochrome A (EA) against different stages of C. albicans biofilms in vitro by XTT reduction assay and crystal violet staining. The mechanism of action of EA on C. albicans biofilm was analyzed with flow cytometry, confocal laser microscopy, and the Real-Time Quantitative Reverse Transcription PCR (qRT-PCR). EA-mediated PACT significantly reduced the viability of C. albicans, with an inhibition rate on biofilm of 89.38% under a concentration of 32 µg/mL EA. We found that EA could not only inhibit the adhesion of C. albicans in the early stage of biofilm formation, but that it also had good effects on pre-formed mature biofilms with a clearance rate of 35.16%. It was observed that EA-mediated PACT promotes the production of a large amount of reactive oxygen species (ROS) in C. albicans and down-regulates the intracellular expression of oxidative-stress-related genes, which further disrupted the permeability of cell membranes, leading to mitochondrial and nuclear damage. These results indicate that EA has good photodynamic antagonizing activity against the C. albicans biofilm, and potential clinical value.

Distribution and diversity of myxomycetes in Tiantangzhai National Forest Park, China.

Although myxomycetes are ubiquitous in terrestrial ecosystems, studies on their distribution and diversity in subtropical humid forests are still lacking. Field collections and moist chamber cultures were conducted from May to October within a two-year period in the Tiantangzhai National Forest Park of China. A total of 1,492 records representing 73 species belonging to 26 genera were obtained, of which 243 records/37 species were from field collections, and 1,249 records/52 species were from moist chamber cultures. Among the specimens obtained by culturing, 896 records/38 species and 353 records/37 species were obtained from living bark and ground litter, respectively. ANOVA showed that the sampling months had significant impacts on collection of myxomycetes from field and those that inhabit litter. An LEfSe analysis indicated that Arcyria was significantly abundant in August, while Stemonitis and Physarum were more abundant in July when collected from field. An RDA analysis showed that temperature was the main factor that affected the litter-inhabiting myxomycetes. The ANOVA indicated that forest type was the significant factor for bark-inhabiting myxomycetes. Diderma effusum was primarily obtained from mixed forests, while Clastoderma debaryanum and Colloderma oculatum were more common in coniferous forests. The RDA analysis indicated that the vegetation, pH, water retention, and elevation were the primary factors that affected the bark-inhabiting myxomycetes.

Nitric oxide regulates perylenequinones biosynthesis in Shiraia bambusicola S4201 induced by hydrogen peroxide.

Shiraia bambusicola has been used as a traditional Chinese medicine for a long history. Its major medicinal active metabolites are perylenequinones, including hypocrellin A, elsinochrome A and so on. At present, the fermentation yield of perylenequinones is low, and its complex biosynthesis and regulatory pathways are still unclear. In this study, nitric oxide, as a downstream signal molecule of hydrogen peroxide, regulates the biosynthesis of perylenequinones. Exogenous addition of 0.01 mM sodium nitroprusside (nitric oxide donor) can promote perylenequinones production by 156% compared with the control. Further research found that hydrogen peroxide and nitric oxide increased the transcriptional level of the biosynthetic genes of hypocrellin A. The results showed that nitric oxide is involved in the biosynthesis and regulation of perylenequinones in Shiraia bambusicola as a signal molecule. In the future, the yield of perylenequinones can be increased by adding exogenous nitric oxide in fermentation.

Differentiated mineral nutrient management in two bamboo species under elevated CO2 environment.

Mineral nutrients play a critical role in maintaining plant growth, but are vulnerable to climate change, such as elevated atmospheric carbon dioxide (CO2) concentrations. Previous studies reported that impact of elevated CO2 concentrations on plant growth vary among plant species, which may affect differential mineral nutrient cycling among plant species. However, little is known about how increasing CO2 concentrations affect mineral nutrient uptake and allocation in bamboo species. Using open top chambers (OTCs), we investigated the effects of elevated CO2 concentrations on three key mineral nutrients (iron (Fe), calcium (Ca), and magnesium (Mg)) in two mature bamboo species (Phyllostachys edulis and Oligostachyum lubricum). Results showed increased leaf and root biomass under elevated CO2 concentrations (P. edulis: 30.24% and 10.94%; O. lubricum: 24.47% and 13.84%, respectively). Conversely, elevated CO2 concentrations had negligible effects on the biomass of other bamboo organs (e.g., branches and culms). To a certain extent, elevated CO2 concentrations also caused nutrient variation among the various organs of these two species. For Ph. edulis, elevated CO2 concentrations increased mineral content (Fe, Ca, and Mg) in and allocation to leaves while it decreased Fe and Mg allocation to roots. By contrast, elevated CO2 concentrations only increased mineral content in and allocation to O. lubricum leaves and decreased Mg to its roots. Results confirmed that elevated CO2 concentrations resulted in differential mineral nutrient uptake and allocation response between these two species. Understanding such differences is critical to the sustainable nutrient management of bamboo ecosystems under increasing CO2 concentrations.

Genome Sequencing and Analysis of the Hypocrellin-Producing Fungus Shiraia bambusicola S4201.

Shiraia bambusicola has long been used as a traditional Chinese medicine and its major medicinal active metabolite is hypocrellin, which exhibits outstanding antiviral and antitumor properties. Here we report the 32 Mb draft genome sequence of S. bambusicola S4201, encoding 11,332 predicted genes. The genome of S. bambusicola is enriched in carbohydrate-active enzymes (CAZy) and pathogenesis-related genes. The phylogenetic tree of S. bambusicola S4201 and nine other sequenced species was constructed and its taxonomic status was supported (Pleosporales, Dothideomycetes). The genome contains a rich set of secondary metabolite biosynthetic gene clusters, suggesting that strain S4201 has a remarkable capacity to produce secondary metabolites. Overexpression of the zinc finger transcription factor zftf, which is involved in hypocrellin A (HA) biosynthesis, increases HA production when compared with wild type. In addition, a new putative HA biosynthetic pathway is proposed. These results provide a framework to study the mechanisms of infection in bamboo and to understand the phylogenetic relationships of S. bambusicola S4201. At the same time, knowledge of the genome sequence may potentially solve the puzzle of HA biosynthesis and lead to the discovery of novel genes and secondary metabolites of importance in medicine and agriculture.

Identification and Nematicidal Characterization of Proteases Secreted by Endophytic Bacteria Bacillus cereus BCM2.

The endophytic bacterium Bacillus cereus BCM2 has shown great potential as a biocontrol organism against Meloidogyne incognita, which causes severe root-knot diseases in crops. In our previous study, the metabolite of BCM2 showed high nematicidal activity against the M. incognita second-stage juveniles. However, the mechanism employed by endophytic bacteria to infect and kill nematodes is still unclear. Here, we investigate both the endophytic bacterial extracellular proteins with nematicidal activity and their mechanism of killing nematodes. The first step was detecting the nematicidal activities of crude proteins. The results show that the nematode mortality rate reached 100% within 72 h, and the crude proteins damaged both the cuticle and eggshell, before finally destroying the targets. This suggests possible proteinaceous pathogeny in BCM2. Throughout the process, the fine-detail changes in the nematode cuticle and the intestinal structure were observed using scanning electron microscopy and transmission electron microscopy. These images show that BCM2 extracellular proteins did not damage the internal organization of the nematode but did severely damage its cuticle, which led to content leakage. From the crude proteins, chitosanase, alkaline serine protease, and neutral protease were purified and identified. The M. incognita-B. cereus BCM2 microenvironment simulation demonstrates that BCM2 adheres to the surface of nematodes and helps the metabolites that were produced by BCM2 to rapidly recognize and kill M. incognita. This relationship between plants, endophytic bacteria, and nematodes offers insight into the biological mechanisms that can be utilized for of nematode management.

Evaluation of hypocrellin A-loaded lipase sensitive polymer micelles for intervening methicillin-resistant Staphylococcus Aureus antibiotic-resistant bacterial infection.

There is an urgent need for new antibacterial strategies to overcome the emergence of antibiotic resistance. Antibacterial photodynamic therapy (APDT) may be an effective method to deliver photosensitizers for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections. Here, we report that the photosensitizer hypocrellin A (HA) loaded into lipase-sensitive methoxy poly (ethylene glycol)-block-poly(ε-caprolactone) (mPEG-PCL) micelles showed high anti-MRSA activity in vitro and in vivo by PDT. Once the micelles come into contact with bacteria that secrete lipase, the PCL is degraded to release HA. Our results showed that the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of mPEG-PCL/HA micelles after light irradiation were 0.69 and 1.38 mg/L (HA concentration), respectively. In the dark, the MIC and MBC of the micelles were 250 and 500 mg/L (HA concentration), respectively. The fluorescent stain results further demonstrated the photodynamic antibacterial activity of mPEG-PCL/HA micelles. The survival rate of mice subjected to experimental acute peritonitis increased to 86% after treated with the micelles. The polymeric micelles showed low hemolytic activity and biocompatibility, simultaneously preventing aggregation in vivo and enhancing the water solubility of HA. Thus, the photosensitizer HA loaded micelles could be used as APDT for infections caused by bacteria without antibiotic resistance.

Diaporone A, a new antibacterial secondary metabolite from the plant endophytic fungus Diaporthe sp.

Diaporone A (1), one new dihydroisocoumarin derivative and four known α-dibenzopyrones, alternariol (2), 5'-hydroxyalternariol (3), alternariol 4,10-dimethyl ether (4), and alternariol 4-methyl ether (5) were isolated from the crude extract of the plant endophytic fungus Diaporthe sp. Their structures were determined on the basis of spectroscopic analysis, including 1D and 2D NMR techniques as well as HRESIMS and comparison with data from the literature. The absolute configuration of 1 was assigned by electronic circular dichroism (ECD) calculations. Compound 1 showed moderate antibacterial activity against Bacillus subtilis with the MIC value of 66.7 µM, and exhibited weak cytotoxicity against human cervical carcinoma (HeLa) cell line with IC50 value of 97.4 µM.

A four-locus phylogeny of rib-stiped cupulate species of Helvella (Helvellaceae, Pezizales) with discovery of three new species.

Helvella species are ascomycetous macrofungi with saddle-shaped or cupulate apothecia. They are distributed worldwide and play an important ecological role as ectomycorrhizal symbionts. A recent multi-locus phylogenetic study of the genus suggested that the cupulate group of Helvella was in need of comprehensive revision. In this study, all the specimens of cupulate Helvella sensu lato with ribbed stipes deposited in HMAS were examined morphologically and molecularly. A four-locus phylogeny was reconstructed using partial sequences of the heat shock protein 90, nuclear rDNA internal transcribed spacer region 2, nuclear large subunit ribosomal DNA and translation elongation factor 1-α genes. Three clades were revealed in Helvella sensu stricto. Twenty species were included in the analysis, of which 13 are distributed in China. Three new species, H. acetabuloides, H. sichuanensis and H. tianshanensis, are described and illustrated in detail. A neotype was designated for H. taiyuanensis. Helvella calycina is a new record for China, while Dissingia leucomelaena should be excluded from Chinese mycota. Hsp90 and ITS2 are recommended as useful supplementary barcodes for species identifications of the genus.

Effects of the Endophytic Bacteria Bacillus cereus BCM2 on Tomato Root Exudates and Meloidogyne incognita Infection.

Root-knot nematodes (Meloidogyne spp.) cause serious crop losses worldwide. The colonization of tomato roots by endophytic bacteria Bacillus cereus BCM2 can greatly reduce Meloidogyne incognita damage, and tomato roots carrying BCM2 were repellent to M. incognita second-stage juveniles (J2). Here, the effects of BCM2 colonization on the composition of tomato root exudates was evaluated and potential mechanisms for BCM2-mediated M. incognita control explored using a linked twin-pot assay and GC-MS. On water agar plates, J2 preferentially avoided filter paper treated with tomato root exudates (organic phase only) from plants inoculated with BCM2, visiting these 67.1% less than controls. In a linked twin-pot assay, BCM2 treatment resulted in a 42.0% reduction in the number of nematodes in the soil, a 43.3% reduction in the number of galls and a 47.7% decrease in the density of M. incognita in root tissues. Analysis of root exudate composition revealed that BCM2 inoculation increased the number of components in exudates. Among these, 2,4-di-tert-butylphenol, 3,3-dimethyloctane, and n-tridecane secretions markedly increased. In repellency trials on water agar plates, J2 avoided 2,4-di-tert-butylphenol, n-tridecane, and 3,3-dimethyloctane at concentrations of 4 mmol/liter. In a linked twin-pot assay, inoculation with 2,4-di-tert-butylphenol or 3,3-dimethyloctane reduced the number of nematodes in the soil (by 54.9 and 70.6%, respectively), the number of galls (by 53.7 and 52.4%), and the number of M. incognita in root tissues (by 67.5 and 36.3%). BCM2 colonization in tomato roots affected the composition of root exudates, increasing the secretion of substances that appear to be repellent, thus decreasing M. incognita J2 infection of roots.