1.Effect of VvLaeA on the growth and development of Beauveria bassiana.
Chinese Journal of Biotechnology 2023;39(2):685-694
It is unclear how VvLaeA functions in regulating the growth and development of Volvariella volvacea (Bull. ex. Fr.) Sing.. Firstly, bioinformatics analysis of VvLaeA was carried out in this study. Subsequently, the Vvgpd promoter and the open reading frame (ORF) fragment of VvlaeA were amplified and fused by polymerase chain reaction (PCR). The fusion fragment was cloned into the pK2 (bar) plasmid. The recombinant construct pK2(bar)-OEVvlaeA was transfected into Beauveria bassiana by Agrobacterium tumefaciens-mediated transformation. Finally, the growth and development of the transformants were examined. The results showed that VvLaeA shared a low homology with similar proteins in other fungi. Compared with the wild type, the colony diameter of the transformant was significantly increased. However, the pigment deposition, conidial yields and germination rates were significantly decreased. The overexpression strains were more sensitive to stresses than that of the wild type. Further studies showed the conidial cell wall properties of the transformants were altered, and the expressions of genes related to the conidial development were significantly down-regulated. Collectively, VvLaeA increased the growth rate of B. bassiana strains and negatively regulated the pigmentation and conidial development, which shed a light for the functional identification of straw mushroom genes.
Beauveria/metabolism*
;
Spores, Fungal
;
Growth and Development
2.Lean strategy for data mining and continuous improvement of Chinese pharmaceutical process: a case study of sporoderm-removal Ganoderma lucidum spore powder.
Yi ZHONG ; Xiao-Hui FAN ; Zhen-Hao LI
China Journal of Chinese Materia Medica 2023;48(3):829-834
In the digital transformation of Chinese pharmaceutical industry, how to efficiently govern and analyze industrial data and excavate the valuable information contained therein to guide the production of drug products has always been a research hotspot and application difficulty. Generally, the Chinese pharmaceutical technique is relatively extensive, and the consistency of drug quality needs to be improved. To address this problem, we proposed an optimization method combining advanced calculation tools(e.g., Bayesian network, convolutional neural network, and Pareto multi-objective optimization algorithm) with lean six sigma tools(e.g., Shewhart control chart and process performance index) to dig deeply into historical industrial data and guide the continuous improvement of pharmaceutical processes. Further, we employed this strategy to optimize the manufacturing process of sporoderm-removal Ganoderma lucidum spore powder. After optimization, we preliminarily obtained the possible interval combination of critical parameters to ensure the P_(pk) values of the critical quality properties including moisture, fineness, crude polysaccharide, and total triterpenes of the sporoderm-removal G. lucidum spore powder to be no less than 1.33. The results indicate that the proposed strategy has an industrial application value.
Bayes Theorem
;
Data Mining
;
Drug Industry
;
Powders
;
Reishi
;
Spores, Fungal
3.Isolation, identification, and pathogenicity research of brown rot pathogens from Gastrodia elata.
Xin TANG ; Jin-Qiang ZHANG ; Wei-Ke JIANG ; Qing-Song YUAN ; Yan-Hong WANG ; Lan-Ping GUO ; Yang YANG ; Ye YANG ; Tao ZHOU
China Journal of Chinese Materia Medica 2022;47(9):2288-2295
Brown rot is a common disease in the cultivation and production of Gastrodia elata, but its pathogens have not been fully revealed. In this study, the pathogenic fungi were isolated and purified from tubers of 77 G. elata samples with brown rot. Pathogens were identified by the pathogenicity test and morphological and molecular identification. The pathogenicity of each pathogen and its inhibitory effects on Armillaria gallica were compared. The results showed that 119 strains of fungi were isolated from tubers of G. elata infected with brown rot. Among them, the frequency of separation of Ilyonectria fungi was as high as 42.01%. The pathogenicity test showed that the pathogenicity characteristics of six strains of fungi were consistent with the natural symptoms of brown rot in G. elata. The morphological and molecular identification results showed that the six strains belonged to I. cyclaminicola and I. robusta in the Nectriaceae family of Sordariomycetes class, respectively. Both types of fungi could produce pigments, conidia, and chlamycospore, and the growth rate of I. cyclaminicola was significantly higher than that of I. robusta. The comparison of pathogenicity showed that the spots formed by I. cyclaminicola inoculation were significantly larger than those of I. robusta inoculation, suggesting I. cyclaminicola was superior to I. robusta in pathogenicity. The results of confrontation culture showed that I. cyclaminicola and I. robusta could signi-ficantly inhibit the germination and cordage growth of A. gallica. A. gallica also inhibited the growth of pathogens, and I. cyclaminicola was less inhibited as compared with I. robusta. The results of this study revealed for the first time that I. cyclaminicola and I. robusta were the pathogens responsible for G. elata brown rot.
Fungi
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Gastrodia
;
Plant Tubers
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Spores, Fungal
;
Virulence
4.Identification, biological characterization, and fungicide screening of pathogens causing leaf spot of Belamcanda chinensis.
Ya-Wen YANG ; Qiao-Huan CHEN ; Jia ZHOU ; Bi-Sheng HUANG ; Yu-Huan MIAO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2022;47(22):6042-6049
The leaf spot of Belamcanda chinensis often appears in May to June and spreads rapidly during the flowering stage(July to September) in the cultivation fields, seriously affecting the yield and quality of B. chinensis. To identify and characterize the pathogens of the leaf spot, we isolated two species of Alternaria, identified them according to Koch's postulates, and tested their pathogenicity and biological characteristics. Furthermore, we determined the inhibitory effects of 6 chemical fungicides, 1 plant fungicide, and 3 microbial fungicides on the pathogens by using mycelial growth rate and plate confrontation method to select the appropriate control agents. The results showed that the two pathogens causing B. chinensis leaf spot were Alternaria tenuissima and A. alternata. The conidia of A. tenuissima often formed long chains with no or a few branches, while those of A. alternata often formed short branched chains. The optimum growth temperature of both A. tenuissima and A. alternata was 25 ℃. The two pathogens grew well in alkaline environment. The indoor fungicide screening experiments showed that 40% flusilazole had good inhibitory effects on the two pathogens, with the EC_(50) values of 12.42 mg·L~(-1) and 12.78 mg·L~(-1) for A. tenuissima and A. alternata, respectively. The results of this study provide a theoretical basis for the subsequent theoretical research and field control of B. chinensis leaf spot.
Fungicides, Industrial/pharmacology*
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Research
;
Iris Plant
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Spores, Fungal
;
Mycelium
5.Construction and application of actin fluorescent marker in Verticillium dahliae Kleb.
Bin CHEN ; Juan TIAN ; Zhidi FENG ; Huan WANG ; Meilan LI ; Zhaosheng KONG
Chinese Journal of Biotechnology 2019;35(8):1520-1528
Actin filaments play an important role in fungal life processes such as growth, development and cytokinesis. The expression vector pSULPH-Lifeact-mCherry of fluorescent mCherry-labeled actin was transferred into Verticillium dahliae Kleb. wild type V592 by the genetic transformation system mediated by Agrobacterium tumefaciens to obtain the stable fluorescent labeled actin strain V592/Lifeact-mCherry. Then we detected its biological phenotype and the dynamic changes of actin fluorescence during the process of spore germination, mycelial growth and development. There was no significant difference in the colony morphology, colonial growth rate, sporulation and germination rate between the fluorescent labeled actin strain and the wild type. The actin fluorescence signal was observed at the tip of the conidia and hyphae and the septum clearly. Actin participated in the formation of the contractile actomyosin ring (CAR) during cytokinesis by observing the dynamic behavior of the actin in the process of hyphal septum formation. The fluorescent labeled actin strain can be used to study the dynamics of actin in fungal development to provide theoretical and practical support for further study of the mechanism of actin in fungal development and pathogenesis.
Actins
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Agrobacterium tumefaciens
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Plant Diseases
;
Spores, Fungal
;
Verticillium
6.Efficacy of Diphenyleneiodonium Chloride (DPIC) Against Diverse Plant Pathogens
Boknam JUNG ; Taiying LI ; Sungyeon JI ; Jungkwan LEE
Mycobiology 2019;47(1):105-111
Many of the fungicides and antibiotics currently available against plant pathogens are of limited use due to the emergence of resistant strains. In this study, we examined the effects of diphenyleneiodonium chloride (DPIC), an inhibitor of the superoxide producing enzyme NADPH oxidase, against fungal and bacterial plant pathogens. We found that DPIC inhibits fungal spore germination and bacterial cell proliferation. In addition, we demonstrated the potent antibacterial activity of DPIC using rice heads infected with the bacterial pathogen Burkholderia glumae which causes bacterial panicle blight (BPB). We found that treatment with DPIC reduced BPB when applied during the initial flowering stage of the rice heads. These results suggest that DPIC could serve as a new and useful antimicrobial agent in agriculture.
Agriculture
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Anti-Bacterial Agents
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Burkholderia
;
Cell Proliferation
;
Flowers
;
Germination
;
Head
;
NADPH Oxidase
;
Plants
;
Spores, Fungal
;
Superoxides
7.Alternaria yunnanensis sp. nov., a New Alternaria Species Causing Foliage Spot of Rubber Tree in China
Zhi Ying CAI ; Yi Xian LIU ; Yu Ping SHI ; Li Ming DAI ; Lan Lan LI ; Hong Jun MU ; Mei Lin LV ; Xiao Yong LIU
Mycobiology 2019;47(1):66-75
A new species of Alternaria causing leaf spots on the rubber tree (Hevea brasiliensis) in Yunnan, China, was isolated, examined, and illustrated. Morphologically, it belongs to the section Porri of Alternaria, which produces relatively large conidia and a simple or branched, filamentous long beak. It is, however, characterized by conidiophores gradually enlarging near the apex into a clavate conidiogenous cell and long ellipsoid to obclavate, smooth-walled conidia with a long filamentous beak. Molecular phylogenetic analyses based on ITS rDNA, GAPDH, and TEF1-alpha sequences demonstrate that the phytopathogen falls in the clade of the section Porri, being most closely related to A. sidae, A. sennae, A. deseriticola, A. cyamopsidis, A. rostellata, A. nitrimali, A. crassa, and A. thunbergiae.
Acanthaceae
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Accidental Falls
;
Alternaria
;
Animals
;
Ascomycota
;
Beak
;
China
;
Classification
;
DNA, Ribosomal
;
Hevea
;
Rubber
;
Spores, Fungal
8.Cladophialophora lanosa sp. nov., a New Species Isolated from Soil
Kallol DAS ; Seung Yeol LEE ; Hee Young JUNG
Mycobiology 2019;47(2):173-179
Cladophialophora is characterized by simple hyphomycetes with brown hyphae that give rise to branched chains of pale brown conidia and shows affinities with the Herpotrichiellaceae. A fungal strain belonging to the genus Cladophialophora was isolated from soil in Daegu, Korea. This strain produces numerous greenish to dark black lanose aerial mycelia with hair like structures. It is morphological similar to C. chaetospira, C. inabaensis, and C. multiseptata; however, the conidiophores and conidia sizes of the newly isolated strain (KNU16-032) are clearly different from them. The novelty of the strain was also confirmed based on phylogenetic analysis using the data sets of the internal transcribed spacer region of and the partial sequence of 28S ribosomal DNA region along with the cultural characteristics. Because morphological observations and phylogenetic analysis indicated that the strain is distinct from previously known Cladophialophora species, we propose this species as a new species Cladophialophora lanosa sp. nov., and provide the detailed descriptions in this study.
Cultural Characteristics
;
Daegu
;
Dataset
;
DNA, Ribosomal
;
Hair
;
Hyphae
;
Korea
;
Mitosporic Fungi
;
Soil
;
Spores, Fungal
9.Colletotrichum fructicola, a Member of Colletotrichum gloeosporioides sensu lato, is the Causal Agent of Anthracnose and Soft Rot in Avocado Fruits cv. “Hass”.
Dionicio FUENTES-ARAGÓN ; Sandra Berenice JUÁREZ-VÁZQUEZ ; Mateo VARGAS-HERNÁNDEZ ; Hilda Victoria SILVA-ROJAS
Mycobiology 2018;46(2):92-100
The filamentous Ascomycota Colletotrichum gloeosporioides sensu lato is a fungus that has been reported worldwide as a causal agent of anthracnose disease in avocado and other crops. In Mexico, this species affects fruits from an early stage of development in the orchard until the post-harvest stage. Although fungicides are continuously applied to control Colletotrichum species, pericarp cankers and soft rot mesocarp in fruits are still frequently observed. Considering the lack of a precise description of the causative agent, the aim of the current study was to determine the pathogens involved in this symptomatology. Twenty-four isolates were consistently obtained from the pericarp of avocado fruits cv. “Hass” collected in the central avocado-producing area of Mexico. Morphological features such as colony growth, conidia size, and mycelial appressorium were assessed. Bayesian multilocus phylogenetic analyses were performed using amplified sequences of the internal transcribed spacer region of the nuclear ribosomal DNA; actin, chitin synthase, glyceraldehyde-3-phosphate dehydrogenase partial genes; and APn2-Mat1-2 intergenic spacer and mating type Mat1-2 partial gene from the nine selected isolates. In addition, fruits were inoculated with a conidial suspension and reproducible symptoms confirmed the presence of Colletotrichum fructicola in this area. This pathogenic species can now be added to those previously reported in the country, such as C. acutatum, C. boninense, C. godetiae, C. gloeosporioides, and C. karstii. Disease management programs to reduce the incidence of anthracnose should include C. fructicola to determine its response to fungicides that are routinely applied, considering that the appearance of new species is affecting the commercial quality of the fruits and shifting the original population structure.
Actins
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Ascomycota
;
Chitin Synthase
;
Colletotrichum*
;
Disease Management
;
DNA, Ribosomal
;
Fruit*
;
Fungi
;
Incidence
;
Mexico
;
Oxidoreductases
;
Persea*
;
Spores, Fungal
10.Allergic bronchopulmonary mycosis – pathophysiology, histology, diagnosis, and treatment
Koichiro ASANO ; Katsuhiko KAMEI ; Akira HEBISAWA
Asia Pacific Allergy 2018;8(3):e24-
Allergic bronchopulmonary mycosis (ABPM) develops mainly in patients with asthma or cystic fibrosis via types I and III hypersensitivity reactions to filamentous fungi. Aspergillus spp., especially Aspergillus fumigatus, is the major causative fungus because of its small conidia, thermophilic hyphae, and ability to secrete serine proteases. The cardinal histological feature of ABPM is allergic (eosinophilic) mucin-harboring hyphae in the bronchi, for which the formation of extracellular DNA trap cell death (ETosis) of eosinophils induced by viable fungi is essential. Clinically, ABPM is characterized by peripheral blood eosinophilia, increased IgE levels in the serum, IgE and IgG antibodies specific for fungi, and characteristic radiographic findings; however, there are substantial differences in the clinical features of this disease between East and South Asian populations. Systemic corticosteroids and/or antifungal drugs effectively control acute diseases, but recurrences are quite common, and development of novel treatments are warranted to avoid adverse effects and emergence of drug-resistance due to prolonged treatment with corticosteroids and/or antifungal drugs.
Acute Disease
;
Adrenal Cortex Hormones
;
Antibodies
;
Asian Continental Ancestry Group
;
Aspergillus
;
Aspergillus fumigatus
;
Asthma
;
Bronchi
;
Cell Death
;
Cystic Fibrosis
;
Diagnosis
;
Eosinophilia
;
Eosinophils
;
Extracellular Traps
;
Fungi
;
Humans
;
Hypersensitivity
;
Hyphae
;
Immunoglobulin E
;
Immunoglobulin G
;
Invasive Pulmonary Aspergillosis
;
Recurrence
;
Serine Proteases
;
Spores, Fungal

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