Hyeokjun Yoon

Plant growth promotion and Penicillium citrinum.

BackgroundEndophytic fungi are known plant symbionts. They produce a variety of beneficial metabolites for plant growth and survival, as well as defend their hosts from attack of certain pathogens. Coastal dunes are nutrient deficient and offer harsh, saline environment for the existing flora and fauna. Endophytic fungi may play an important role in plant survival by enhancing nutrient uptake and producing growth-promoting metabolites such as gibberellins and auxins. We screened roots of Ixeris repenes (L.) A. Gray, a common dune plant, for the isolation of gibberellin secreting endophytic fungi.ResultsWe isolated 15 endophytic fungi from the roots of Ixeris repenes and screened them for growth promoting secondary metabolites. The fungal isolate IR-3-3 gave maximum plant growth when applied to waito-c rice and Atriplex gemelinii seedlings. Analysis of the culture filtrate of IR-3-3 showed the presence of physiologically active gibberellins, GA1, GA3, GA4 and GA7 (1.95 ng/ml, 3.83 ng/ml, 6.03 ng/ml and 2.35 ng/ml, respectively) along with other physiologically inactive GA5, GA9, GA12, GA15, GA19, GA20 and, GA24. The plant growth promotion and gibberellin producing capacity of IR-3-3 was much higher than the wild type Gibberella fujikuroi, which was taken as control during present study. GA5, a precursor of bioactive GA3 was reported for the first time in fungi. The fungal isolate IR-3-3 was identified as a new strain of Penicillium citrinum (named as P. citrinum KACC43900) through phylogenetic analysis of 18S rDNA sequence.ConclusionIsolation of new strain of Penicillium citrinum from the sand dune flora is interesting as information on the presence of Pencillium species in coastal sand dunes is limited. The plant growth promoting ability of this fungal strain may help in conservation and revegetation of the rapidly eroding sand dune flora. Penicillium citrinum is already known for producing mycotoxin citrinin and cellulose digesting enzymes like cellulase and endoglucanase, as well as xylulase. Gibberellins producing ability of this fungus and the discovery about the presence of GA5 will open new aspects of research and investigations.

Whole genome and global gene expression analyses of the model mushroom Flammulina velutipes reveal a high capacity for lignocellulose degradation.

Flammulina velutipes is a fungus with health and medicinal benefits that has been used for consumption and cultivation in East Asia. F. velutipes is also known to degrade lignocellulose and produce ethanol. The overlapping interests of mushroom production and wood bioconversion make F. velutipes an attractive new model for fungal wood related studies. Here, we present the complete sequence of the F. velutipes genome. This is the first sequenced genome for a commercially produced edible mushroom that also degrades wood. The 35.6-Mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC42780. The 88.4-kb mitochondrial genome contained 35 genes. Well-developed wood degrading machinery with strong potential for lignin degradation (69 auxiliary activities, formerly FOLymes) and carbohydrate degradation (392 CAZymes), along with 58 alcohol dehydrogenase genes were highly expressed in the mycelium, demonstrating the potential application of this organism to bioethanol production. Thus, the newly uncovered wood degrading capacity and sequential nature of this process in F. velutipes, offer interesting possibilities for more detailed studies on either lignin or (hemi-) cellulose degradation in complex wood substrates. The mutual interest in wood degradation by the mushroom industry and (ligno-)cellulose biomass related industries further increase the significance of F. velutipes as a new model.

Cadophora malorum Cs‐8‐1 as a new fungal strain producing gibberellins isolated from Calystegia soldanella

Fourteen endophytic fungi with different colony morphologies were isolated from the roots of Calystegia soldanella. Endophytic fungi isolated from C. soldanella were identified by internal transcribed spacer (ITS) region. To verify plant growth promotion (PGP), culture filtrates of isolated endophytic fungi were treated in Waito‐c rice (WR) and C. soldanella seedlings. Culture filtrates of Cs‐8‐1 fungal strain had advanced PGP activity. The presence of physiologically bioactive gibberellins (GA) GA1 (1.213 ng ml−1), GA3 (1.292 ng ml−1), GA4 (3.6 ng ml−1), GA7 (1.328 ng ml−1), other inactive GA9 (0.796 ng ml−1) and GA12 (0.417 ng ml−1), GA20 (0.302 ng ml−1), GA24 (1.351 ng ml−1), GA34 (0.076 ng ml−1), and GA53 (0.051 ng ml−1) in culture filtrates of Cs‐8‐1 fungal strain was detected. The Cs‐8‐1 fungal strain was confirmed as a producer of GAs. Molecular analysis of sequences showed high similarity of 99% to Cadophora malorum. Consequentially, the Cs‐8‐1 fungal strain was identified as a new C. malorum producing GAs.

Diversity and Plant Growth-Promotion of Endophytic Fungi Isolated from the Roots of Plants in Dokdo Islands

Endophytic fungi were isolated from the roots of plants growing naturally on the island of Dokdo. Plant samples, such as Miscanthus sinensis, Achyranthus japonica and Echinochloa crusgali were isolated from Dongdo, and those such as Honkenya peploides and Artemsia koidzumii were isolated from Seodo. Twenty one strains of endophytic fungi were isolated from these plants. To identify the strains, PCR (polymerase chain reaction) amplification of the partial ITS (Internal Transcribed Spacer) regions was done with universal primers ITS-1 and ITS-4 to determine the nucleotide sequence of the ITS regions. Of the strains isolated from Miscanthus sinensis, 75% were Penicillium sp. and 25% were Aspergillus sp. Fifty five percent of strains isolated from Achyranthus japonica were Penicillium sp., 30% were Aspergillus sp. and 15% were Zygorhynchus sp. Strains isolated from Echinochloa crusgali were Penicillium sp. (50%), Aspergillus sp. (12%), Giberella sp. (13%), Talaromyces sp. (9%) and Umbelopsis sp. (8%). Of the strains isolated from Honkenya peploides, 76% were Penicillium sp. and 24% were Pestalotiopsis sp. Strains isolated from Artemisia koidzumii were Penicillium sp. (81%) and Mucor sp. (19%). As a result of bioassay, Ec-3-1 strain isolated from Echinochloa crusgalli showed plant growth-promotion activity. Of all the endophytic fungi isolated, Penicillium sp. was the most abundantly distributed fungal strain in all plants used in this study.

Comparative analysis of bacterial diversity and communities inhabiting the fairy ring of Tricholoma matsutake by barcoded pyrosequencing

Comparative analysis of the soil bacterial communities inhabiting the fairy ring of Tricholoma matsutake.

Diversity and Plant Growth Promoting Capacity of Endophytic Fungi Associated with Halophytic Plants from the West Coast of Korea

Abstract Five halophytic plant species, Suaeda maritima, Limonium tetragonum, Suaeda australis, Phragmites australis, and Suaeda glauca Bunge, which are native to the Muan salt marsh of South Korea, were examined for fungal endophytes by sequencing the internal transcribed spacer (ITS) region containing ITS1, 5.8S rRNA, and ITS2. In total, 160 endophytic fungal strains were isolated and identified from the roots of the 5 plant species. Taxonomically, all 160 strains belonged to the phyla Ascomycota, Basidiomycota, and Zygomycota. The most dominant genus was Fusarium, followed by the genera Penicillium and Alternaria. Subsequently, using 5 statistical methods, the diversity indices of the endophytes were determined at genus level. Among these halophytic plants, P. australis was found to host the greatest diversity of endophytic fungi. Culture filtrates of endophytic fungi were treated to Waito-C rice seedlings for plant growth-promoting effects. The fungal strain Su-3-4-3 isolated from S. glauca Bunge provide the maximum plant length (20.1 cm) in comparison with wild-type Gibberella fujikuroi (19.6 cm). Consequently, chromatographic analysis of the culture filtrate of Su-3-4-3 showed the presence of physiologically active gibberellins, GA1 (0.465 ng/mL), GA3 (1.808 ng/mL) along with other physiologically inactive GA9 (0.054 ng/mL) and GA24 (0.044 ng/mL). The fungal isolate Su-3-4-3 was identified as Talaromyces pinophilus.

Culturable Fungal Endophytes Isolated from the Roots of Coastal Plants Inhabiting Korean East Coast

Abstract Twelve plant species were collected from the east coast of Korea to identify culturable endophytes present in their roots. The fungal internal transcribe spacer (ITS) region (ITS1-5.8SrRNA-ITS2) was used as a DNA barcode for identification of fungi. A total of 194 fungal strains were identified and categorized into 31 genera. The genus Penicillium accounted for the largest number of strains, followed by the genus Aspergillus. Furthermore, using 5 statistical methods, the diversity indices of the fungi were calculated at the genus level. After comprehensive evaluation, the endophytic fungal group from Phragmites australis ranked highest in diversity analyses. Several strains responsible for plant growth and survival (Penicillium citrinum, P. funiculosum, P. janthinellum, P. restrictum, and P. simplicissimum), were also identified. This study provides basic data on the sheds light on the symbiotic relationship between coastal plants and fungi.

Diversity of Endophytic Fungi from the Roots of Halophytes Growing in Go-chang Salt Marsh

Halophyte samples, such as Suaeda japonica, Phragmites australis, Limonium tetragonum, Suaeda maritima were collected from Go-chang salt marsh. Thirty-nine endophytic fungal strains were isolated from the roots of halophytes naturally growing in salt marsh. All endophytic fungal strains isolated were analyzed by internal transcribed spacer (ITS) containing ITS1, 5.8 s and ITS2 region. Endophytic fungal strains belong to eight orders, i.e., Eurotiales (36%), Pleosporales (26%), Hypocreales (18%), Incertae sedis (8%), Glomerellales (5%), Sordariales (2%), Xylariales (2%), and Capnodiales (3%). On genus level, they were composed of Alternaria, Aspergillus, Chaetomium, Cladosporium, Colletotrichum, Coniothyrium, Dothideomycete, Fusarium, Gibberella, Macrophoma, Penicillium, Pestalotiopsis, Phaeosphaeria, Phoma, Pleosporales, Pseudozyma, Talaromyces, and Termitomyces. Of them, Penicillium (26%), Fusarium (13%) of Eurotiales and Hypocreales were predominant.

Metagenomic analysis of soil fungal communities on Ulleungdo and Dokdo Islands.

Ulleungdo and Dokdo are volcanic islands that experience a characteristic marine climate, influenced by warm currents. The richness and diversity of the plant species, particularly vascular plants, are higher on Ulleungdo than on Dokdo. In contrast to the native plant life, little is known about the diversity of soil fungi living in the rhizosphere of these two islands. In this study, we utilized the barcoded pyrosequencing method to analyze rhizosphere soil fungi on Ulleungdo and Dokdo. In total, 768 operational taxonomic units (OTUs) were analyzed from the Ulleungdo samples, while 640 OTUs and 382 OTUs were analyzed from the Dongdo and Seodo (islets of Dokdo) samples, respectively. Species richness was considerably higher in the Ulleungdo samples than in the Dongdo and Seodo samples, while there was little difference in species diversity between the samples. The taxonomic composition analyses demonstrated that members of the phylum Basidiomycota dominated the Ulleungdo samples, whereas members of the phylum Ascomycota were predominant in the Dokdo samples. Ectomycorrhizal fungi belonging to the phylum Basidiomycota, in particular, were more abundant in the Ulleungdo samples. This finding suggests that the difference in the abundance of the ectomycorrhizal fungi in the rhizospheres of Ulleungdo and Dokdo may have been affected by species richness and diversity of the vascular plants. Our study is the first detailed report of the composition of soil fungal communities on the Ulleungdo and Dokdo islands. In addition, our findings provide a basis for understanding the ecological interactions between plants and fungi.

Plant Growth-Promoting Activity and Genetic Diversity of Endophytic Fungi Isolated from Native Plants in Dokdo Islands for Restoration of a Coastal Ecosystem

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.