• Journal of Microbiology and Biotechnology
• /
• v.30 no.7
• /
• pp.1027-1036
• /
• 2020

Stipa purpurea is a unique and dominant herbaceous plant species in the alpine steppe and meadows on the Qinghai-Tibet Plateau (QTP). In this work, we analyzed the composition and diversity of the culturable endophytic fungi in S. purpurea according to morphological and molecular identification. Then, we investigated the bioactivities of these fungi against plant pathogenic fungi and 1-aminocyclopropane-1-carboxylate deaminase (ACCD) deaminase activities. A total of 323 fungal isolates were first isolated from S. purpurea, and 33 fungal taxa were identified by internal transcribed spacer primers and grouped into Ascomycota. The diversity of endophytic fungi in S. purpurea was significantly higher in roots as compared to leaves. In addition, more than 40% of the endophytic fungi carried the gene encoding for the ACCD gene. The antibiosis assay demonstrated that 29, 35, 28, 37 and 34 isolates (43.9, 53.1, 42.4, 56.1, and 51.5%) were antagonistic to five plant pathogenic fungi, respectively. Our study provided the first assessment of the diversity of culture-depending endophytic fungi of S. purpurea, demonstrated the potential application of ACCD activity and antifungal activities with potential benefits to the host plant, and contributed to high biomass production and adaptation of S. purpurea to an adverse environment.

• Journal of Life Science
• /
• v.23 no.1
• /
• pp.95-101
• /
• 2013

In this study, plant samples of five species were collected from the Dokdo islands in South Korea. Plant samples such as Asparagus schoberioides, Corydalis platycarpa, Festuca rubra, Sedum oryzifolium, and Setaria viridis were collected from the Dongdo and Seodo. Endophytic fungal strains were isolated from the roots of five plants from the Dokdo islands. Thirty-three fungal strains were isolated from these native plants. All the endophytic fungi were analyzed by internal transcribed spacer (ITS) sequencing (ITS containing ITS1, 5.8s, and the ITS2 region). Waito-c rice seedlings were treated with fungal culture filtrates to test their plant growth-promoting activity. A bioassay of the D-So-1-1 fungal strain isolated from S. oryzifolium confirmed that it has the highest plant growth-promoting activity. All the endophytic fungi belong to four orders: Eurotiales (86%), Capnodiales (3%), Hypocreales (4%), and Incertae sedis (7%). The endophytic fungi were classified as Ascomycota, which contained Aspergillus (12%), Cladosporium (3%), Eurotium (3%), Fusarium (18%), Microsphaeropsis (6%), and Penicillium (58%) at the genus level.

• Mycobiology
• /
• v.45 no.3
• /
• pp.160-171
• /
• 2017

Larvae of Bradysia agrestis, an insect vector that transports plant pathogens, were sampled from geographically isolated regions in Korea to identify their cutaneous fungal and bacterial flora. Sampled areas were chosen within the distribution range of B. agrestis; each site was more than 91 km apart to ensure geographical segregation. We isolated 76 microbial (fungi and bacteria) strains (site 1, 29; site 2, 29; site 3, 18 strains) that were identified on the basis of morphological differences. Species identification was molecularly confirmed by determination of universal fungal internal transcribed spacer and bacterial 16S rRNA gene sequences in comparison to sequences in the EzTaxon database and the NCBI GenBank database, and their phylogenetic relationships were determined. The fungal isolates belonged to 2 phyla, 5 classes, and 7 genera; bacterial species belonged to 23 genera and 32 species. Microbial diversity differed significantly among the geographical groups with respect to Margalef's richness (3.9, 3.6, and 4.5), Menhinick's index (2.65, 2.46, and 3.30), Simpson's index (0.06, 0.12, and 0.01), and Shannon's index (2.50, 2.17, and 2.58). Although the microbial genera distribution or diversity values clearly varied among geographical groups, common genera were identified in all groups, including the fungal genus Cladosporium, and the bacterial genera Bacillus and Rhodococcus. According to classic principles of co-evolutionary relationship, these genera might have a closer association with their host insect vector B. agrestis than other genera identified. Some cutaneous bacterial genera (e.g., Pseudomonas) displaying weak interdependency with insect vectors may be hazardous to agricultural environments via mechanical transmission via B. agrestis. This study provides comprehensive information regarding the cutaneous microflora of B. agrestis, which can help in the control of such pests for crop management.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.7
• /
• pp.996-1006
• /
• 2019

Objective: This study aimed to gain deeper insights into the dynamic changes in spoilage fungi populations during fermentation and the influence of traditional additives on silage quality. Methods: Elephant grass (Pennisetum purpureum) was prepared without any additive (control), and with the addition of 0.5% salt, and 0.5% salt-0.2% sugar mixture. The fungal community was then determined using a classic culturing method and high-throughput sequencing at 0, 5, 15, and 60 days after ensiling. Results: The results showed that the fungal community of elephant grass silage varied significantly between the natural fermentation without any additive and the two additive groups. The diversity and relative abundance of spoilage molds in the control group were much higher than those in the two treatment groups (p<0.05). Three species of yeasts (Candida sp., Pichia sp., Trichosporon sp.) and four spoilage molds (Fusarium sp., Aspergillus sp., Muco sp. and Penicillin sp.) were the predominant fungi in elephant grass during natural fermentation from 0 to 60 days, which were found to be significantly decreased in salt and sugar additive groups (p<0.05). Meanwhile, the diversity and relative abundance of undesirable molds in the 0.5%-salt additive group were the lowest among all groups. Conclusion: Adding salt and sugar, particularly 0.5% salt, is a promising effective approach to reduce the amount of undesirable fungi thus, improving the silage quality of elephant grass in northern Vietnam.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.1
• /
• pp.101-111
• /
• 2017

The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

• Journal of Microbiology
• /
• v.43 no.6
• /
• pp.475-486
• /
• 2005

Fungal secondary metabolites constitute a wide variety of compounds which either playa vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to playa vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz., phenylalanine ammonia-lyase, cinnamic acid hydroxylase and p-coumarate CoA ligase) in the A. oryzae genome undoubtedly prove the extent of its metabolic diversity. Since many of these genes have not been identified earlier, knowledge on their corresponding products or activities remain undeciphered. In future, it is anticipated that these enzymes may be reasonable targets for metabolic engineering in fungi to produce agriculturally and nutritionally important metabolites.

• Journal of Ginseng Research
• /
• v.36 no.2
• /
• pp.211-217
• /
• 2012

Endophytic fungi were isolated from various tissues (root, stem, petiole, leaf, and flower stalk) of 3- and 4-year-old ginseng plants (Panax ginseng Meyer) cultivated in Korea. The isolated endophytic fungi were identified based on the sequence analysis of the internal transcribed spacer (ITS), 1-5.8-ITS 2. A morphological characterization was also conducted using microscopic observations. According to the identification, 127 fungal isolates were assigned to 27 taxa. The genera of Phoma, Alternaria and Colletotrichum were the most frequent isolates, followed by Fusarium, Entrophospora and Xylaria. Although 19 of the 27 taxa were identified at the species level, the remainder were classified at the genus level (6 isolates), phylum level (Ascomycota, 1 isolate), and unknown fungal species (1 isolate). Endophytic fungi of 13 and 19 species were isolated from 3- and 4-year-old ginseng plants, respectively, and Phoma radicina and Fusarium solani were the most frequently isolated species colonizing the tissues of the 3- and 4-year-old ginseng plants, respectively. The colonization frequency (CF%) was dependant on the age and tissue examined: the CFs of the roots and stems in the 3-year-old ginseng were higher than the CF of tissues in the 4-year-old plants. In contrast, higher CFs were observed in the leaves and petioles of 4-year-old plants, and endophytic fungi in the flower stalks were only detected in the 4-year-old plants. In conclusion, we detected diverse endophytic fungi in ginseng plants, which were distributed differently depending on the age and tissue examined.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• v.18 no.1
• /
• pp.159-159
• /
• 2005

• Journal of Microbiology and Biotechnology
• /
• v.25 no.5
• /
• pp.732-737
• /
• 2015

We present here an in silico search of fungal sterol-esterase/lipase and bacterial depolymerase sequences from environmental metagenomes. Both enzyme types contain the α/β-hydrolase protein fold. Analysis of DNA conserved motifs, protein homology search, phylogenetic analysis, and protein 3D modeling have been used, and the efficiency of these screening strategies is discussed. The presence of bacterial genes in the metagenomes was higher than those from fungi, and the sequencing depth of the metagenomes seemed to be crucial to allow finding enough diversity of enzyme sequences. As a result, a novel putative PHA-depolymerase is described.

• Mycobiology
• /
• v.48 no.1
• /
• pp.44-50
• /
• 2020

Calonectria pseudonaviculata and C. henricotiae are two closely related fungal species responsible for boxwood blight disease of ornamental shrubs (Buxus spp.) in the U.S. and Europe. A previous study has shown isolates of the latter species, which is restricted to Europe, to be less sensitive to tetraconazole, an azole fungicide. In this study, we have analyzed the CYP51 paralogs for polymorphism in 26 genomes, representing geographically disparate populations of C. pseudonaviculata (n = 19) and C. henricotiae (n = 7), from the U.S., Europe, Asia, and New Zealand. The presence of a CYP51A pseudogene and lack of a functional CYP51A paralog in all C. pseudonaviculata genomes examined is a novel discovery for fungi and could have implications for the evolution of resistance to antifungal chemicals.