Epitypification can solve many taxonomic problems and stabilize the understanding of species, genera, families or orders. The aim of this paper is to illustrate how to epitypify. A few examples where taxa have been epitypified are considered and the benefits and disadvantages of epitypification are discussed. We also outline some examples of taxa which need to be epitypified with reasons.
Colletotrichum ; classification ; genetics ; DNA, Fungal ; genetics ; isolation & purification ; Fungi ; classification ; genetics ; Mycological Typing Techniques ; methods ; Phylogeny

In this paper, 29 endophytes were isolated from different organs and tissues of Lycium barbarum of Ningxia by tablet coating method, 18 of them was fungi, and 11 of them was actinomycetes. The endophytes quantity in the different tissues were leaves > flowers > roots >fruits; The hydroxyl radical scavenging activities of 11 endophytes were investigated by Fenton reaction, and total antioxidant capacities of them were examined by a. total antioxidant capacity test kit; culture features and strain-specific sequence analysis were employed to explore the diversity of the 11 endophytes. The result showed that 5 fungi and 6 actinomycetes that having antioxidant activity could be phylogenetically classified into 3 genera, 3 genera and 3 families, respectively. The total antioxidant capacity and hydroxyl radical scavenging activity of the 11 endophytes showed distinct difference. The antioxidant activity of Aspergillus were stronger, among which total antioxidant capacity of fL1 was (188.5 ± 0.549) U · mL⁻¹ and the IC₅₀ was 0.3 mg · L⁻¹; the IC₅₀ of strain fL1 was 0.42 mg · L⁻¹ and the total antioxidant capacity of fL9 was (113.63 ± 1.021) U · mL⁻¹, all of them were stronger than the positive control Vit C. The experimental results indicated that endophytic fungi of L. barbarum of Ningxia have a great developing and application prospect for the development of antioxidant agent.
Antioxidants ; chemistry ; Bacteria ; chemistry ; classification ; genetics ; isolation & purification ; Biodiversity ; Endophytes ; classification ; genetics ; isolation & purification ; Fungi ; chemistry ; classification ; genetics ; isolation & purification ; Lycium ; microbiology ; Molecular Sequence Data ; Oxidation-Reduction

OBJECTIVETo evaluate fungal contamination on the surface of Chinese herbal medicines and explore an appropriate method for fast and efficient identification of contaminant fungi.

METHODChinese herbal medicines were first washed and the washing solution was plated onto potato dextrose agar (PDA) to obtain the pure isolates. For molecular identification, two new pairs of specific primers were designed according to ITS region of fungi genome sequences. The strains were identified through polymerase chain reaction (PCR) and sequence analysis.

RESULTFifty fungal strains were obtained from the surface of 15 Chinese herbal medicines with the percent of contaminated samples of 93.3%. Twenty-seven strains among them were successfully identified.

CONCLUSIONFungal contamination on the surface of Chinese herbal medicines is quite common. Although different fungal species were isolated, the genus Aspergillus was the predominant. The primer pairs developed in this study are compatible and can be used to identify fungal species from the surface of Chinese herbal medicines.

Drug Contamination ; Drugs, Chinese Herbal ; Fungi ; genetics ; isolation & purification ; Polymerase Chain Reaction

OBJECTIVEThe research focused on identification the pathogen of Aquilaria sinensis dieback disease and confirmation the function of inducing agarwood formation by fungi.

METHODThe morphological observation, rDNA ITS sequence analyses, Koch's postulates and pathogenicity test were used to identify the isolates.

RESULTThe isolates of the causal agent was Lasiodiplodia theobromae, which induced the plant to produce highly valuable agarwood.

CONCLUSIONThe first report of dieback disease of Aquilaria sinensis caused by L. theobromae, and our research results laid a theoretical foundation for agarwood production by using fungus.

Fungi ; classification ; genetics ; isolation & purification ; physiology ; Molecular Sequence Data ; Phylogeny ; Plant Diseases ; microbiology ; Thymelaeaceae ; microbiology

The swollen root of Rehmannia glutinosa is used as one kind of important Chinese traditional medicine. The root of R. glutinosa usually swelled in rotational cropping but not in continuous cropping. The rhizosphere microorganisms of R. glutinosa under different farming condition were thought related to that. In this study, the endophytic fungi in the root of R. glutinosa growing in various soil conditions were isolated for the study of the relationship between the microorganisms and the root enlargement of their host plants. The dominant endophytes, Verticillium spp., Fusarium oxysporum, F. redolens and Ceratobasidium spp. were identified by morphological observation and 18S rDNA and ITS sequence analysis. The preliminary investigation showed that the excessive growth of Verticillium and Fusarium genus fungi is unfavorable for the R. glutinosa root swelling, but Ceratobasidium fungi has no effects on the root enlargement.
DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; Fungi ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Plant Roots ; microbiology ; Rehmannia ; microbiology

A total of 24 biologically pure entophytic fungal strains were isolated from stems, leaves, and seed coats of Xylocarpus plants by repeated purification, and identified with Internal Transcribed Spacer (ITS) rDNA molecular method, which belonging to 14 genera, 11 families, 9 orders and 3 classes. There were differences in genus and species levels among three plant materials from different habitats and species, and it was found that the strains of Phomopsis and Colletotrichum existed in all three plant materials. In vitro assay of antitumor activity by MTT method revealed that the EtOAc extracts of 15 strains exhibited potent antitumor activity. These results suggest that it is of value for further investigation on the above fungal strains.
Antineoplastic Agents ; isolation & purification ; pharmacology ; Biodiversity ; Cell Line, Tumor ; Endophytes ; chemistry ; classification ; genetics ; isolation & purification ; Fungi ; chemistry ; classification ; genetics ; isolation & purification ; HCT116 Cells ; Humans ; Meliaceae ; microbiology ; Phylogeny

BACKGROUND: The identification of molds in clinical laboratories is largely on the basis of phenotypic criteria, the classification of which can be subjective. Recently, molecular methods have been introduced for identification of pathogenic molds in clinical settings. Here, we employed comparative sequence analysis to identify molds. METHODS: A total of 47 clinical mold isolates were used in this study, including Aspergillus and Trichophyton. All isolates were identified by phenotypic properties, such as growth rate, colony morphology, and reproductive structures. PCR and direct sequencing, targeting the internal transcribed spacer (ITS) region, the D1/D2 region of the 28S subunit, and the beta-tubulin gene, were performed using primers described previously. Comparative sequence analysis by using the GenBank database was performed with the basic local alignment search tool (BLAST) algorithm. RESULTS: For Aspergillus, 56% and 67% of the isolates were identified to the species level by using ITS and beta-tubulin analysis, respectively. Only D1/D2 analysis was useful for Trichophyton identification, with 100% of isolates being identified to the species level. Performances of ITS and D1/D2 analyses were comparable for species-level identification of molds other than Aspergillus and Trichophyton. In contrast, the efficacy of beta-tubulin analysis was limited to genus identification because of the paucity of database information for this gene. CONCLUSIONS: The molecular methods employed in this study were valuable for mold identification, although the different loci used had variable usefulness, according to mold genus. Thus, a tailored approach is recommended when selecting amplification targets for molecular identification of molds.
Aspergillus/genetics/isolation & purification ; DNA, Fungal/analysis/isolation & purification ; Databases, Genetic ; Fungi/genetics/*isolation & purification ; Humans ; Polymerase Chain Reaction ; RNA, Ribosomal, 28S/*genetics ; Sequence Analysis, DNA ; Trichophyton/genetics/isolation & purification ; Tubulin/*genetics

In order to transform main active ingredient of volatile oil, endophytic fungi were screened from the root of Atractylodes lancea. Transformation method was used in vitro. The changes of volatile oil were traced by gas chromatography. One endophytic fungus (strain ALG-13) which could uitilize volatile oil selectively was screened. Single factor experiment were conducted for exploring the effects of various factors that including kinds of carbon source, speed, liquid volume, pH and concentration of plant tissue on degradation by this strain. Subsequently, the main affecting factors carbon source, speed, pH and liquid volume were optimized using orthogonal array design. Results showed that endophytic fungus ALG-13 selectively used the volatile oil, change the relative percentage of the main components of volatile oil, Atractylon and Atractydin were increased, While, beta-eudesmol and Atractylol decreased. After selectively degradation by fungus, volatile oil components percentage were closer to the geo-herbs. Strain ALG-13 was identified as Bionectria ochroleuca according to its morphological characteristics and systematic analysis of ITS sequence. The optimal conditions were as follows: sucrose used as carbon source, rotating speed was 200 r x min(-1), initial pH for medium was 4.5, 50 mL liquid was added in 250 mL flask. The endophytic fungus ALG-13 could degrade the volatile oil selectively, which was benefit for forming geoherbs A. lancea volatile oil composition.
Atractylodes ; chemistry ; microbiology ; Fungi ; classification ; genetics ; isolation & purification ; metabolism ; Oils, Volatile ; chemistry ; metabolism ; Phylogeny ; RNA, Ribosomal, 18S ; genetics

OBJECTIVETo find the development rules of microbial community in rhizoma sphere of the cultivated Atractylodes lancea.

METHODTotal bacteria, fungi and actinomyces were counted by CFU x g(-1) though dilution plate method. And genomic DNA of microbes were extracted and amplified by primers of E. coli's 27f and 1492r to get the 16S rDNA, then the restriction endonuclease Hinf was used to digest the 16S rDNA.

RESULTTotal bacteria, fungi and actinomyces in 2-year old soil were lower than in 1-year old soil, they decreased 46. 14%, 49. 25%, 31.88% respectively and made the ratio of themselves changed. At the same time, all the 8 soil samples got fine 16S rDNA bands, which were about 1500 bp. And the main bands of most of the samples were found at 1000 bp, but the weak bands of each were different although most bands in the same year samples were more similar than in different year ones.

CONCLUSIONIt is indicated that the change of soil microbial community may has some relation to the continous cropping barrier of A. lancea.

Actinomyces ; genetics ; isolation & purification ; Atractylodes ; growth & development ; Bacteria ; genetics ; isolation & purification ; Biodiversity ; Colony Count, Microbial ; DNA, Ribosomal ; genetics ; Fungi ; genetics ; isolation & purification ; Plants, Medicinal ; growth & development ; RNA, Ribosomal, 16S ; genetics ; Rhizome ; growth & development ; Soil Microbiology

In this paper, the varying pattern of the amount of rhizospheric microorganisms, including bacteria, actinomycetes and fungus, was observed during the cultivation of Paris polyphylla var. yunnanensis. And the correlations between number of rhizospheric microorganisms and the quality of P. polyphylla var. yunnanensis were also studied. The results showed that the rhizospheric microorganism source of P. polyphylla var. yunnanensis was rich. The distribution of rhizospheric microorganisms (soil bacteria, fungus, actinomycetes, potassium-solubilizing bacteria, inorganic phosphorus-solubilizing bacteria, organic phosphorus-solubilizing bacteria) collected from different origin places existed significant difference (P < 0.05). The varying pattern for the amount of rhizospheric microorganisms was showed as following: the amount of bacteria > the amount of actinomycetes > the amount of fungus. The medicinal quality of P. polyphylla var. yunnanensis was influenced by their habits, and the increase of cultivation years caused the obvious decrease of the quality of P. polyphylla var. yunnanensis. Therefore, the increase of cultivation years will cause the variation of the soil micro-ecology flora, and decrease the nutrient absorption and the utilization of P. polyphylla var. yunnanensis, which will make the decrease of the medical quality of P. polyphylla var. yunnanensis.
Bacteria ; genetics ; growth & development ; isolation & purification ; Biodiversity ; China ; Fungi ; genetics ; growth & development ; isolation & purification ; Liliaceae ; chemistry ; microbiology ; Plant Extracts ; analysis ; Rhizome ; chemistry ; microbiology ; Rhizosphere ; Saponins ; analysis ; Soil Microbiology