Analysis of Risk Factors for Pulmonary Infections During Radiotherapy in Lung Cancer Patients.

Objective: To investigate the risk factors for lung infection in lung cancer patients undergoing radiotherapy. Methods: We selected 142 patients with lung cancer who underwent radiotherapy at our hospital from January 2020 to June 2021. The patients were divided into groups according to whether they had pulmonary infection during radiotherapy in our hospital, which was infected group (n=44) and the uninfected group (n=98), respectively. To observe the incidence of lung infection in lung cancer patients during radiotherapy. The distribution of pathogenic bacteria in patients with pulmonary infection was observed. Clinical data of the two groups were collected and compared. The risk factors of lung cancer patients complicated with lung infection were analyzed by binary Logistic regression. Results: All patients with lung cancer complicated with lung infection underwent relevant examination, and the results showed that they were all complicated infections, and the composition ratio of Klebsiella pneumoniae was the highest (31.82%), followed by Staphylococcus, Pseudomonas, and fungi, which accounted for 27.27%, 22.73%, and 18.18%, respectively. Binary Logistic regression analysis showed that age ≥60 years old, smoking history ≥30 years, radiotherapy duration of combined drug regimen > 2 weeks, pathogenic bacteria combined infection, albumin content < 30 g/L were risk factors for lung cancer patients during radiotherapy. Conclusion: Age ≥60 years old, smoking history ≥30 years old, radiotherapy duration of combined drug regimen > 2 weeks, pathogenic bacteria combined infection, albumin content < 30 g/L are the risk factors for lung cancer patients during radiotherapy. Clinical prevention and intervention should be based on the aforementioned independent risk factors to decrease the incidence of lung infections, thereby enhancing patient prognosis.

The catalase-peroxidase PiCP1 plays a critical role in abiotic stress resistance, pathogenicity and asexual structure development in Phytophthora infestans.

Catalase-peroxidase is a heme oxidoreductase widely distributed in bacteria and lower eukaryotes. In this study, we identified a catalase-peroxidase PiCP1 (PITG_05579) in Phytophthora infestans. PiCP1 had catalase/peroxidase and secretion activities and was highly expressed in sporangia and upregulated in response to oxidative and heat stresses. Compared with wild type, PiCP1-silenced transformants (STs) had decreased catalase activity, reduced oxidant stress resistance and damped cell wall integrity. In contrast, PiCP1-overexpression transformants (OTs) demonstrated increased tolerance to abiotic stresses and induced the upregulation of PR genes in the host salicylic acid pathway. The high concentration of PiCP1 can also induced callose deposition in plant tissue. Importantly, both STs and OTs have severely reduced sporangia formation and zoospore releasing rate, but the sporangia germination rate and type varied depending on environmental conditions. Comparative sequence analyses show that catalase-peroxidases are broadly distributed and highly conserved among soil-borne plant parasitic oomycetes, but not in freshwater-inhabiting or strictly plants-inhabiting oomycetes. In addition, we found that silencing PiCP1 downregulated the expression of PiCAT2. These results revealed the important roles of PiCP1 in abiotic stress resistance, pathogenicity and in regulating asexual structure development in response to environmental change. Our findings provide new insights into catalase-peroxidase functions in eukaryotic pathogens.

Characterization of Cetobacterium somerae CPU-CS01 isolated from the intestine of healthy crucian carp (Carassius auratus) as potential probiotics against Aeromonas hydrophila infection.

Cetobacterium somerae is a commensal bacterium for many fish species. However, research on C. somerae has been limited so far, and its function and beneficial potential require to be further investigated. The objective of this study was to evaluate the probiotic properties of C. somerae CPU-CS01 isolated from the intestinal contents of crucian carp (Carassius auratus). Hemolytic activity, antibiotic susceptibility, acid tolerance, bile salt tolerance, free radical scavenging, and enzyme production properties were tested for in vitro. Caenorhabditis elegans and zebrafish (Danio rerio) model were used to evaluate the antioxidant and anti-infective effects of C. somerae CPU-CS01 in vivo. Our results showed that C. somerae CPU-CS01 had no hemolytic activity, it produced cellulase, amylase, and survived at low pH (2.0-3.0) and in the presence of bile salts. The cell-free culture supernatant (CFCS) of C. somerae CPU-CS01 possessed DPPH radical, hydroxyl radical, and superoxide anion scavenging activity. C. elegans fed with C. somerae CPU-CS01 were more resistant to hydrogen peroxide-induced oxidative stress and Aeromonas hydrophila infection. In addition, zebrafish-fed diets containing C. somerae CPU-CS01 showed improved survival after A. hydrophila infection. Based on these results, the positive probiotic properties of C. somerae CPU-CS01 isolated from the intestinal contents of crucian carp make it a potential candidate for probiotic.

Effects of Transcranial Direct Current Stimulation and Neuromuscular Joint Facilitation on Upper Limb Motor Disorders for Stroke Patients.

Context: Approximately 1.5- to 2-million new patients suffer from stroke annually in China. 60% of patients suffering from stroke will sustain different degrees of upper limb dysfunction at six months after onset. Recovery of upper limb function after stroke is of great significance in improving patients' quality of life. Objective: The study intended to explore the rehabilitative the effects of transcranial direct current stimulation combined with neuromuscular joint therapy on the rehabilitation of patients with upper-limb motor disorders after strokes to provide new ideas for rehabilitative treatment. Design: The study was a paired control test. Setting: The study took place in the Department of Rehabilitation Medicine at Heping Hospital of Changzhi Medical College in Changzhi, Shanxi, China. Participants: Participants were 80 stroke patients with upper-limb motor disorders who were treated at the hospital between January 2020 and December 2020. Intervention: According to the natural grouping method, the research team divided participants into an intervention group (n = 42) and a control group (n = 38). The control group received transcranial direct-current stimulation, and the intervention group received transcranial direct-current stimulation combined with neuromuscular joint therapy. Outcome Measures: The measurements included the scores on the Fugl-Meyer Assessment (FMA) scale, the Action Research Arm Test (ARAT), activities of daily living (ADL), and National Institutes of Health Stroke Scale (NIHSS) as well as the serum levels of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and superoxide dismutase (SOD). The team also measured the maximum isometric torque of flexion and extension of the elbow joint. The research team compared the differences in the scores between the groups for all variables. Results: Postintervention, the FMA, ARAT, and ADL scores, the torques of elbow flexion and extension maximum isometric contraction, the amplitude, and the serum BDNF, NGF, and SOD levels were significantly higher in the intervention group than those in the control group, while the NIHSS score and the incubation period of evoked potential were significantly lower than those in the control group. Conclusions: Transcranial direct current stimulation combined with the neuromuscular joint method demonstrated good rehabilitative effects on upper-limb movement disorders for stroke patients and significantly improved their upper-limb function and promoted recovery of nerve functions.

SCFAs improve disease resistance via modulate gut microbiota, enhance immune response and increase antioxidative capacity in the host.

To evaluate the effects of dietary short chain fatty acids (SCFAs) on the intestinal health and innate immunity in crucian carp, a six-week feeding trial was carried out with following treatments: basal diet (BD), basal diet supplementation with 1% sodium acetate (BDSA), basal diet supplementation with 1% sodium propionate (BDSP) and basal diet supplementation with 1% sodium butyrate (BDSB). The results showed dietary BDSA, BDSP and BDSB could protect the host against oxidative stress by improving the activity of certain antioxidative enzymes (T-SOD, GSH-Px and CAT). Additionally, dietary SCFAs could enhance mucosal and humoral immune responses by improving certain innate immune parameters in serum and skin mucus productions (IgM, ACH50 and T-SOD). Furthermore, dietary BDSA and BDSP could up-regulate the expression of immune related genes (TNF-α, TGF-ß and IL-8) and tight junction protein genes (occludin and ZO-1). Dietary BDSB could also elevate the expression of IL-8, TGF-ß, ZO-1 and Occludin in the midgut. Although dietary differences of SCFAs didn't alter the α-diversity of the intestinal flora, they altered the core microbiota. Finally, the challenge trial showed that dietary basal diet supplementation with SCFAs could protect zebrafish against Aeromonas hydrophila. These results suggest that dietary SCFAs could improve innate immunity, modulate gut microbiota and increase disease resistance in the host, which indicated the potential of SCFAs as immunostimulants in aquaculture.

Test-Retest Reliability of a New Device Versus a Long-Arm Goniometer to Evaluate Knee Proprioception.

CONTEXT: Many methods used to evaluate knee proprioception have shortcomings that limit their use in clinical settings. Based on an inexpensive 3D camera, a new portable device was recently used to evaluate the joint position sense (JPS) of the knee joint. However, the test-retest reliability of the new method remains unclear. This study aimed to evaluate the test-retest reliability of the new device and a long-arm goniometer for assessing knee JPS, and to compare the variability of the 2 methods. DESIGN: Prospective observational study of the test-retest reliability of knee JPS measurements. METHODS: Twenty-one healthy adults were tested in 2 sessions with a 1-week interval. Three target knee flexion angles (30°, 45°, and 60°) were reproduced in each session. Target and reproduced angles were measured with both methods. Intraclass correlation coefficients, standard error of the measurement, and Bland-Altman plots were used to quantify test-retest reliability. Paired t tests were used to compare knee JPS (absolute error of the target-reproduced angle) between the methods. RESULTS: The new device (good to excellent intraclass correlation coefficients .74-.80; standard error of the measurement 0.52°-0.61°) demonstrated better test-retest reliability than the goniometer (poor to fair intraclass correlation coefficients .23-.43; standard error of the measurement 0.89°-2.07°) and better test-retest agreement (respective mean differences for the 30°, 45°, and 60° knee angles: 0.11°, 0.13°, and 0.41° for the new system; 0.84°, 1.52°, and 1.18° for the goniometer). The measurements (absolute errors of the target-reproduced angles) with the goniometer were significantly greater than those with the new device (P < .05); the SDs of repeated measurements with the goniometer (1.50°-2.41°) were greater than with the new device (1.08°-1.38°). CONCLUSIONS: Given that the new device has good reliability and sufficient precision, it is the better alternative for evaluating knee JPS. Goniometers should be used with caution to assess knee JPS.

The Cutinase Bdo_10846 Play an Important Role in the Virulence of Botryosphaeria dothidea and in Inducing the Wart Symptom on Apple Plant.

Botryosphaeria dothidea is a pathogen with worldwide distribution, infecting hundreds of species of economically important woody plants. It infects and causes various symptoms on apple plants, including wart and canker on branches, twigs, and stems. However, the mechanism of warts formation is unclear. In this study, we investigated the mechanism of wart formation by observing the transection ultrastructure of the inoculated cortical tissues at various time points of the infection process and detecting the expression of genes related to the pathogen pathogenicity and plant defense response. Results revealed that wart induced by B. dothidea consisted of proliferous of phelloderm cells, the newly formed secondary phellem, and the suberized phelloderm cells surrounding the invading mycelia. The qRT-PCR analysis revealed the significant upregulation of apple pathogenesis-related and suberification-related genes and a pathogen cutinase gene Bdo_10846. The Bdo_10846 knockout transformants showed reduced cutinase activity and decreased virulence. Transient expression of Bdo_10846 in Nicotiana benthamiana induced ROS burst, callose formation, the resistance of N. benthamiana to Botrytis cinerea, and significant upregulation of the plant pathogenesis-related and suberification-related genes. Additionally, the enzyme activity is essential for the induction. Virus-induced gene silencing demonstrated that the NbBAK1 and NbSOBIR1 expression were required for the Bdo_10846 induced defense response in N. benthamiana. These results revealed the mechanism of wart formation induced by B. dothidea invasion and the important roles of the cutinase Bdo_10846 in pathogen virulence and in inducing plant immunity.

Candidate Effectors from Botryosphaeria dothidea Suppress Plant Immunity and Contribute to Virulence.

Fungal effectors play important roles in host-pathogen interactions. Botryosphaeria dothidea is an ascomycetous fungus that is responsible for the diseases of hundreds of woody plant species, including apple ring rot, which seriously affects apples worldwide. However, little is known about the effectors of B. dothidea. In this study, we analyzed the B. dothidea genome and predicted 320 candidate effector genes, 124 of which were successfully amplified and cloned. We investigated the effects of these genes on plant cell death in Nicotiana benthamiana while using a transient expression system. Twenty-four hours after initial inoculation with Agrobacterium tumefaciens cells carrying candidate effectors, the infiltrated leaves were challenged with A. tumefaciens cells carrying the BAX gene. In total, 116 candidate effectors completely inhibited, while one partially inhibited, the programmed cell death (PCD) of N. benthamiana induced by BAX, whereas seven candidate effectors had no effect. We then further tested seven candidate effectors able to suppress BAX-triggered PCD (BT-PCD) and found that they all completely inhibited PCD triggered by the elicitors INF1, MKK1, and NPK1. This result suggests that these effectors were activated in order to suppress pathogen-associated molecular pattern-triggered immunity. The signal peptides of these candidate effectors exhibited secretory activity in yeast (pSUC2 vector). Moreover, the respective deletion of Bdo_11198 and Bdo_12090 significantly reduced the virulence of B. dothidea. These results suggest that these effectors play important roles in the interaction of B. dothidea with its hosts.

An efficient gene disruption method for the woody plant pathogen Botryosphaeria dothidea.

BACKGROUND: Botryosphaeria dothidea causes apple white rot and infects many tree plants. Genome data for B. dothidea are available and many pathogenesis-related genes have been predicted. However, a gene manipulation method is needed to study the pathogenic mechanism of B. dothidea. RESULTS: We established a gene disruption (GD) method based on gene homologous recombination (GHR) for B. dothidea using polyethylene glycol-mediated protoplast transformation. The results showed that a GHR cassette gave much higher GD efficiency than a GHR plasmid. A high GD efficiency (1.3 ± 0.14 per 106 protopasts) and low frequency of random insertions were achieved with a DNA cassette quantity of 15 µg per 106 protoplasts. Moreover, we successfully disrupted genes in two strains. Bdo_05381-disrupted transformants produced less melanin, whereas the Bdo_02540-disrupted transformant showed a slower growth rate and a stronger resistance to Congo red. CONCLUSION: The established GD method is efficient and convenient and has potential for studying gene functions and the pathogenic mechanisms of B. dothidea and other coenocytic fungi.

Efficacy and Safety of 1% Atropine on Retardation of Moderate Myopia Progression in Chinese School Children.

Background: To evaluate the long-term efficacy and safety of topical 1% atropine for retarding moderate myopia. Methods: A randomized, controlled study evaluating atropine and placebo in 660 Chinese children. Patients received drops q1month for 24 months, then q2month for 12 months, followed by no drops for 12 months. Spherical equivalent, axial length, intraocular pressure and atropine-related side effects were examined at 6, 12, 24, 36 and 48 months for all children. Results: Spherical equivalent, myopic progression, axial length augmentation, and progression rate were significantly reduced in the atropine group than those in the placebo group (all P<0.05), indicating that 1% atropine effectively retarded myopia. Moreover, myopic rebound and adverse effects of 1% atropine were eliminated by gradual withdrawal and elimination of 1% atropine. Furthermore, pupil size, near visual acuity, and amplitude of accommodation returned to pretreatment levels after withdrawal of atropine. Conclusion: Topical 1% atropine periodically and alternatively in phase I with gradual reduction in phase II and final withdrawal in phase III may effectively improve atropine efficacy, retard moderate myopia, reduce atropine side effects, minimize myopic rebound, and increase compliance of children simultaneously.

Isolation, Identification of Carotenoid-Producing Rhodotorula sp. from Marine Environment and Optimization for Carotenoid Production.

Carotenoids are natural pigments found in plants and microorganisms. These important nutrients play significant roles in animal health. In contrast to plant production, the advantages of microbial fermentation of carotenoids are the lower media costs, fast growth rate of microorganisms, and the ease of culture condition control. In this study, a colony of red pigment-producing yeast, Rhodotorula sp. RY1801, was isolated from the sediment of marine environment with the potential to produce carotenoids. Optimization of carotenoid production in Rhodotorula sp. RY1801 was also discussed. The optimum conditions found for carotenoid production were as follows: temperature, 28 °C; pH 5.0; carbon source, 10 g/L glucose, nitrogen source, 10 g/L yeast extract, maximum concentration of 987 µg/L of total carotenoids was obtained. The results of this study show that the isolated yeast strain Rhodotorula sp. RY1801 can potentially be used in future as a promising microorganism for the commercial production of carotenoids.

Recombinant N-acyl homoserine lactone-Lactonase AiiAQSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp.

Quorum quenching (QQ) is a promising alternative infection-control strategy to antibiotics that controls quorum-regulated virulence without killing the pathogens. Aeromonas hydrophila is an opportunistic gram-negative pathogen living in freshwater and marine environments. A. hydrophila possesses an N-acyl homoserine lactone (AHL)-based quorum-sensing (QS) system that regulates virulence, so quorum signal-inactivation (i.e., QQ) may represent a new way to combat A. hydrophila infection. In this study, an AHL lactonase gene, aiiA was cloned from Bacillus sp. strain QSI-1 and expressed in Escherichia coli strain BL21(DE3). The A. hydrophila hexanoyl homoserine lactone (C6-HSL) QS signal molecule was degraded by AiiAQSI-1, which resulted in a decrease of bacterial swimming motility, reduction of extracellular protease and hemolysin virulence factors, and inhibited the biofilm formation of A. hydrophila YJ-1 in a microtiter assay. In cell culture studies, AiiAQSI-1 decreased the ability of A. hydrophila adherence to and internalization by Epithelioma papulosum cyprini (EPC) cells. During in vivo studies, oral administration of AiiAQSI-1 via feed supplementation attenuated A. hydrophila infection in Crucian Carp. Results from this work indicate that feed supplementation with AiiAQSI-1 protein has potential to control A. hydrophila aquaculture disease via QQ.

Detection and Identification of Six Monilinia spp. Causing Brown Rot Using TaqMan Real-Time PCR from Pure Cultures and Infected Apple Fruit.

Brown rot is a severe disease affecting stone and pome fruit. This disease was recently confirmed to be caused by the following six closely related species: Monilinia fructicola, M. laxa, M. fructigena, Monilia polystroma, M. mumecola, and M. yunnanensis. Because of differences in geographic distributions, some of these species are important quarantine pathogens in certain countries. In this study, we developed TaqMan real-time polymerase chain reaction (PCR) assays to detect and identify the six species. Primer pairs and probes were designed for Monilinia fructicola, M. fructigena, M. laxa, and Monilia polystroma based on sequence differences in the laccase-2 genes. Additionally, based on sequence differences in the elongation factor genes, primer pairs and probes were designed for Monilia mumecola and M. yunnanensis. The real-time PCR assays were able to specifically identify the target pathogens, with detection limits of 10 to 100 fg of DNA, which is equivalent to one to seven conidia. The assays were also able to detect the target pathogens in a mixed DNA sample comprising all six Monilinia spp. and related species. The real-time PCR assays accurately detected target fungi from infected apple fruit. Furthermore, the identification results were consistent with those of traditional morphological methods.

Characterization of a novel strain of Botryosphaeria dothidea chrysovirus 1 from the apple white rot pathogen Botryosphaeria dothidea.

A novel strain of Botryosphaeria dothidea chrysovirus 1 was identified. It encodes a shortened RNA-dependent RNA polymerase and an elongated coat protein, and it might cause hypovirulence of the host fungal strain.

Diversity, evolution and expression profiles of histone acetyltransferases and deacetylases in oomycetes.

BACKGROUND: Oomycetes are a group of fungus-like eukaryotes with diverse microorganisms living in marine, freshwater and terrestrial environments. Many of them are important pathogens of plants and animals, causing severe economic losses. Based on previous study, gene expression in eukaryotic cells is regulated by epigenetic mechanisms such as DNA methylation and histone modification. However, little is known about epigenetic mechanisms of oomycetes. RESULTS: In this study, we investigated the candidate genes in regulating histone acetylation in oomycetes genomes through bioinformatics approaches and identified a group of diverse histone acetyltransferases (HATs) and histone deacetylases (HDACs), along with three putative novel HATs. Phylogenetic analyses suggested that most of these oomycetes HATs and HDACs derived from distinct evolutionary ancestors. Phylogenetic based analysis revealed the complex and distinct patterns of duplications and losses of HATs and HDACs in oomycetes. Moreover, gene expression analysis unveiled the specific expression patterns of the 33 HATs and 11 HDACs of Phytophthora infestans during the stages of development, infection and stress response. CONCLUSIONS: In this study, we reveal the structure, diversity and the phylogeny of HATs and HDACs of oomycetes. By analyzing the expression data, we provide an overview of the specific biological stages of these genes involved. Our datasets provide useful inputs to help explore the epigenetic mechanisms and the relationship between genomes and phenotypes of oomycetes.

miRNA-451 inhibits glioma cell proliferation and invasion by downregulating glucose transporter 1.

MicroRNAs play an important role in tumor development and progression. Tumor growth is closely associated with glucose metabolism. Specifically, tumor cells produce energy (ATP) under aerobic and anaerobic conditions through glycolysis and metabolites, such as lactic acid and ATP, as a result of the Warburg effect. However, the transport of glucose into cells depends on protein transporters in the cell membrane. Therefore, this area has recently become a topic of interest for research on targeted cancer therapy. We found that miRNA-451 inhibits the phosphatidylinositol-3 kinase (PI3K)/Akt signaling pathway to modify the biological behavior of glioma cells. Inhibiting the PI3K/Akt pathway may prevent glucose-addicted cancer cells from performing glycolysis. Akt directly affects glycolysis by regulating the localization of the glucose transporter 1 (GLUT1). However, how miRNA-451 regulates glucose transporters on the cell membrane and affects the regulatory mechanisms of glucose metabolism in glioma cells remains unclear. Consequently, we predict and verify related gene protein interactions. By targeting CAB 39, miRNA-451 likely triggers the LKB1/AMPK/PI3K/AKT pathway, which regulates GLUT1, to inhibit the glucose metabolism of, reduce the energy supply to, and inhibit the proliferation and invasion of glioma cells. Our results suggest a new direction for the treatment of glioma.

Monilinia Species Associated with Brown Rot of Cultivated Apple and Pear Fruit in China.

Monilinia isolates were collected from major apple and pear production regions in China from 2004 to 2011 and identified based on their morphological characteristics and three highly conserved loci. The 247 isolates belonged to three species: Monilinia fructicola, Monilia yunnanensis, and Monilia polystroma. M. yunnanensis was the most prevalent (77%), followed by M. polystroma (20%) and Monilinia fructicola (3%). Monilia yunnanensis is primarily distributed in the south, north, and west of China; M. polystroma is limited to the north and east; and Monilinia fructicola was detected only from a few samples from the north and east. Phylogenetic analysis based on internal transcribed spacer, ß-tubulin, and laccase (lcc2) genes suggested that Monilia yunnanensis, M. polystroma, and Monilinia fructigena are closely related, and Monilia yunnanensis is more distantly related. We also found that these three species do not show consistent differences in morphological characteristics, including colony morphology, colony expansion rate, conidial characteristics, and the amount of stroma produced in culture. Thus, these three species are more like phylogenetic species in the process of speciation. In addition, a set of species-specific primers based on single-nucleotide polymorphisms and deletions in the lcc2 gene region were designed and a conventional polymerase chain reaction method successfully developed for differentiating Monilinia fructicola, Monilia yunnanensis, M. polystroma, and Monilinia laxa from the other species.

Genomic and transcriptomic analysis of the endophytic fungus Pestalotiopsis fici reveals its lifestyle and high potential for synthesis of natural products.

BACKGROUND: In recent years, the genus Pestalotiopsis is receiving increasing attention, not only because of its economic impact as a plant pathogen but also as a commonly isolated endophyte which is an important source of bioactive natural products. Pestalotiopsis fici Steyaert W106-1/CGMCC3.15140 as an endophyte of tea produces numerous novel secondary metabolites, including chloropupukeananin, a derivative of chlorinated pupukeanane that is first discovered in fungi. Some of them might be important as the drug leads for future pharmaceutics. RESULTS: Here, we report the genome sequence of the endophytic fungus of tea Pestalotiopsis fici W106-1/CGMCC3.15140. The abundant carbohydrate-active enzymes especially significantly expanding pectinases allow the fungus to utilize the limited intercellular nutrients within the host plants, suggesting adaptation of the fungus to endophytic lifestyle. The P. fici genome encodes a rich set of secondary metabolite synthesis genes, including 27 polyketide synthases (PKSs), 12 non-ribosomal peptide synthases (NRPSs), five dimethylallyl tryptophan synthases, four putative PKS-like enzymes, 15 putative NRPS-like enzymes, 15 terpenoid synthases, seven terpenoid cyclases, seven fatty-acid synthases, and five hybrids of PKS-NRPS. The majority of these core enzymes distributed into 74 secondary metabolite clusters. The putative Diels-Alderase genes have undergone expansion. CONCLUSION: The significant expansion of pectinase encoding genes provides essential insight in the life strategy of endophytes, and richness of gene clusters for secondary metabolites reveals high potential of natural products of endophytic fungi.

The bZIP transcription factor PfZipA regulates secondary metabolism and oxidative stress response in the plant endophytic fungus Pestalotiopsis fici.

The bZIP transcription factors are conserved in all eukaryotes and play critical roles in organismal responses to environmental challenges. In filamentous fungi, several lines of evidence indicate that secondary metabolism (SM) is associated with oxidative stress mediated by bZIP proteins. Here we uncover a connection with a bZIP protein and oxidative stress induction of SM in the plant endophytic fungus Pestalotiopsis fici. A homology search of the P. fici genome with the bZIP protein RsmA, involved in SM and the oxidative stress response in Aspergillus nidulans, identified PfZipA. Deletion of PfzipA resulted in a strain that displayed resistant to the oxidative reagents tert-butylhydroperoxide (tBOOH), diamide, and menadione sodium bisulfite (MSB), but increased sensitivity to H2O2 as compared to wild type (WT). Secondary metabolite production presented a complex pattern dependent on PfzipA and oxidative reagents. Without oxidative treatment, the ΔPfzipA strain produced less isosulochrin and ficipyroneA than WT; addition of tBOOH further decreased production of iso-A82775C and pestaloficiol M in ΔPfzipA; diamide treatment resulted in equivalent production of isosulochrin and ficipyroneA in the two strains; MSB treatment further decreased production of RES1214-1 and iso-A82775C but increased pestaloficiol M production in the mutant; and H2O2 treatment resulted in enhanced production of isosulochrin, RES1214-1 and pestheic acid but decreased ficipyroneA and pestaloficiol M in ΔPfzipA compared to WT. Our results suggest that PfZipA regulation of SM is modified by oxidative stress pathways and provide insights into a possible role of PfZipA in mediating SM synthesis in the endophytic lifestyle of P. fici.

Rupestriomyces and Spissiomyces, two new genera of rock-inhabiting fungi from China.

Bare rocks in arid and semi-arid climates may harbor a bewildering biodiversity of fungi that are overlooked in China. During a survey of rock-inhibiting fungi in China, more than 1000 were isolated and 14 belonging to Dothideomycetes (Ascomycota) were selected for detailed study. Phylogenetic trees based on combined sequence datasets of mt 16S rDNA, partial nuc rDNA 18S and 28S indicated that these strains clustered in two distinct, well supported and previously unknown lineages within the class Dothideomycetes. Therefore two new genera were established corresponding to those two clades. Spissiomyces gen. nov. is characterized by thick-walled, yellowish brown hyphae and globose or subglobose conidia, if present. Rupestriomyces gen. nov. is characterized by globose, barrel-shaped, ampulliform or ovoidal conidia formed from acropetal, catenate hyphae. Further phylogenetic analyses using combined sequence datasets of the rDNA internal transcribed spacer (ITS) region and 16S rDNA, part of genes of RPB2, TUB2, nuc rDNA 18S and 28S revealed that those strains represented five new species (i.e. Spissiomyces aggregatus, S. ramosus, Rupestriomyces sinensis, R. ampulliformis, R. torulosus). They were described, illustrated and compared with similar taxa based on morphological characteristics and phylogenetic relationships. The results of this paper provides insight into the richness and diversity of rock-inhibiting fungi in nature.