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Revisiting Rhytisma lonicericola: Morphological Characterization and Molecular Phylogenetic Analysis

Bok-Nam JUNG ; Ji-Hyun PARK ; Hyeon-Dong SHIN

Mycobiology.2022;50(2):150-154. doi:10.1080/12298093.2022.2054527

Rhytisma lonicericola was identified as a tar spot fungus on Lonicera sp. in 1902, and has since been recorded on several species of Lonicera in China, Japan, and Korea. Most of the previous records of R. lonicericola have been based on a list of disease occurrences in the absence of any formal morphological identification or molecular analyses. Using six newly obtained specimens collected in the past 2 years, we confirmed the tar spot fungus found on L. japonica in Korea as R. lonicericola based on morphological examinations and molecular phylogenetic analyses. This fungus was distinguished from R. xylostei, another tar spot fungus on Lonicera, by ascospore size and geographical distributions. We present detailed mycological information and, for the first time, DNA sequence data useful for the identification of R. lonicericola.

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Exploration of Mycobiota in Cypripedium japonicum, an Endangered Species

Gyeongjun CHO ; Geun-Hye GANG ; Hee-Young JUNG ; Youn-Sig KWAK

Mycobiology.2022;50(2):142-149. doi:10.1080/12298093.2022.2064409

Orchids live with mycorrhizal fungi in mutualism. This symbiotic relationship plays an essential role in the overall life cycle of orchids from germination, growth, settlement, and reproduction. Among the 1000 species of the orchid, the Korean lady’s slipper, Cypripedium japonicum, is known as an endangered species. Currently, only five natural habitats of the Korean lady’s slipper remain in South Korea, and the population of Korean lady’s slipper in their natural habitat is not increasing. To prevent extinction, this study was designed to understand the fungal community interacting in the rhizosphere of the Korean lady’s slipper living in the native and artificial habitats. In-depth analyses were performed to discover the vital mycorrhizal fungi contributing to habitat expansion and cultivation of the endangered orchid species. Our results suggested that Lycoperdon nigrescens contributed most to the increase in natural habitats and Russula violeipes as a characteristic of successful cultivation. And the fungi that helped L. nigrescens and R. violeipes to fit into the rhizosphere community in Korean lady’s slipper native place were Paraboeremia selaginellae and Metarhizium anisopliae, respectively. The findings will contribute to restoring and maintaining the endangered orchid population in natural habitats.

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Mycorrhization of Quercus spp. with Tuber huidongense and T. himalayense Collected in Korea

Ju-Hui GWON ; Hyeok PARK ; Ahn-Heum EOM

Mycobiology.2022;50(2):104-109. doi:10.1080/12298093.2022.2065717

Fungi of the genus Tuber are ectomycorrhizal fungi that form a symbiotic relationship mainly with oak and hazel trees. Tuber spp. exhibit a highly selective host plant preference; thus, for cultivation purposes it is important to select an appropriate host plant for successful mycorrhization. In addition, as mycorrhizal characteristics differ according to Tuber spp., it is necessary to understand the differences in mycorrhizae according to the fungal species. Tuber huidongense and Tuber himalayense were recently discovered in Korea; therefore, we used spore suspensions from these two species to inoculate two species of oak trees, Quercus acutissima and Quercus dentata, to compare colonization rates and morphologies of the mycorrhizae. The colonization rates demonstrated that the different Tuber spp. favored different host plant species. In addition, unique morphological and anatomical characteristics were observed for T. huidongense and T. himalayense depending on the host species. These findings can lead to new economically important agricultural activities related to truffle cultivation in Korea.

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Diaporthe eleutharrhenae sp. nov. Associated with a Critically Endangered Liana in China

Shijie SONG ; Sven LANDREIN

Mycobiology.2022;50(2):99-103. doi:10.1080/12298093.2022.2058164

A new fungus isolated from the leaves of Eleutharrhena macrocarpa in southern Yunnan, China is described using morphological and molecular evidence. Phylogenetic trees based on the combined nuclear ribosomal DNA internal transcribed spacer (ITS), translation elongation factor-1a (TEF1), and b-tubulin gene (TUB2) sequences showed that Diaporthe eleutharrhenae sp. nov. is sister to Diaporthe chinensis N.I. de Silva, Lumyong & K.D. Hyde and morphologically differs in shorter alpha conidia (5–8.5 Â 1.5–2 mm) and the presence of beta conidia. This study also resolves a nomenclatural problem, as two taxa were published using the same name. To avoid confusion, the unrelated D. chinensis H. Dong, J. W. Xia & X. G.Zhang is here renamed as D. dongii (H. Dong, J. W. Xia & X. G. Zhang) S. J. Song & Landrein, sp. nov. in honor of the author that described this species. Study and description of fungi associated with threatened tropical species could help to understand their ecology as well as the potential spread of fungi onto cultivated crop species.

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Effects of Culture Mechanism of Cinnamomum kanehirae and C. camphora on the Expression of Genes Related to Terpene Biosynthesis in Antrodia cinnamomea

Zhang ZHANG ; Yi WANG ; Xiao-Long YUAN ; Ya-Na LUO ; Ma-Niya LUO ; Yuan ZHENG

Mycobiology.2022;50(2):121-131. doi:10.1080/12298093.2022.2059156

The rare edible and medicinal fungus Antrodia cinnamomea has a substantial potential for development. In this study, Illumina HiSeq 2000 was used to sequence its transcriptome.The results were assembled de novo, and 66,589 unigenes with an N50 of 4413 bp were obtained. Compared with public databases, 6,061, 3,257, and 2,807 unigenes were annotated to the Non-Redundant, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes databases, respectively. The genes related to terpene biosynthesis in the mycelia of A. cinnamomea were analyzed, and acetyl CoA synthase (ACS2 and ACS4), hydroxymethylglutaryl CoA reductase (HMGR), farnesyl transferase (FTase), and squalene synthase (SQS) were found to be upregulated in XZJ (twig of C. camphora) and NZJ (twig of C. kanehirae). Moreover, ACS5 and 2,3-oxidized squalene cyclase ( OCS) were highly expressed in NZJ, while heme IX farnesyl transferase (IX-FIT) and ACS3 were significantly expressed in XZJ. The differential expression of ACS1, ACS2, HMGR, IX-FIT, SQS, and OCS was confirmed by real-time quantitative reverse transcription PCR. This study provides a new concept for the additional exploration of the molecular regulatory mechanism of terpenoid biosynthesis and data for the biotechnology of terpenoid production.

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Isolation, Characterization, and Metabolic Profiling of Ceratorhiza hydrophila from the Aquatic Plant Myriophyllum spicatum

Yasmin M. ELSABA ; Arezue BOROUJERDI ; Asmaa ABDELSALAM

Mycobiology.2022;50(2):110-120. doi:10.1080/12298093.2022.2059889

The goal of the present study was to investigate the antibacterial properties, enzyme production, and metabolic profiling of a new Ceratorhiza hydrophila strain isolated from the submerged aquatic plant Myriophyllum spicatum. Furthermore, the fungus’ morphological characterization and DNA sequencing have been described. The fungus has been identified and submitted to the GenBank as Ceratorhiza hydrophila isolate EG19 and the fungus ID is MK387081. The enzyme analyses showed its ability to produce protease and cellulase enzymes. According to the CSLI standard, the ethyl acetate extract of C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia, Micrococcus luteus, and Staphylococcus aureus. Metabolic profiling has been carried out using 700 MHz NMR spectroscopy. Based on the 1 H and 1 H- 13 C heteronuclear single quantum coherence (HSQC) NMR data and NMR databases, 23 compounds have been identified. The identified metabolites include 31% amino acids, 9% sugars, 9% amines, 4% sugar alcohols, and 4% alkaloids. This is the first report for the metabolic characterization of C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia,, which gave preliminary information about the fungus. It is expected that our findings not only will pave the way to other perspectives in enormous applications using C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia, as a new promising source of antimicrobial agents and essential metabolites, but also it will be valuable in the classification and chemotaxonomy of the species.

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Genetic Diversity of Amylomyces rouxii from Ragi tapai in Java Island Based on Ribosomal Regions ITS1/ITS2 and D1/D2

Ega DELVA ; Tuty ARISURYANTI ; Miftahul ILMI

Mycobiology.2022;50(2):132-141. doi:10.1080/12298093.2022.2028436

Amylomyces rouxii is commonly found as amylolytic fungi in tapai fermentation. However, its diversity is rarely reported despite being often used for food production in Southeast Asia. This research aims to analyze the genetic diversity and the distribution pattern of A. rouxii from Ragi tapai in Java Island, Indonesia. We isolated the fungus from samples obtained from Ragi tapai producing centers in Bandung, Sumedang, Muntilan, Blora, Yogyakarta, and Bondowoso. The obtained isolates were molecularly identified based on the ribosomal regions ITS1/ITS2 and D1/D2, then analyzed for phylogenetic tree reconstruction, genetic distance, genetic variation, and haplotype networking. Six isolates showed specific morphological traits ofA. rouxii. However, phylogenetic tree reconstruction on the ribosomal genes showed that the isolates were grouped into two different clades related to two species. Clade A included BDG, SMD, and MTL isolates related to A. rouxii, whereas clade B included YOG, BLR, and BDS isolates related to Mucor indicus. The genetic distances between clades for ITS1/ITS2 and D1/D2 were 0.6145 and 0.1556, respectively. In conclusion, we confirmed the genetic diversity of molds from Ragi tapai in Java Island and showed that the isolates are not only related to A. rouxii as reported before.

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Re-Identification on Korean Penicillium Sequences in GenBank Collected by Software GenMine

Chang Wan SEO ; Sung Hyun KIM ; Young Woon LIM ; Myung Soo PARK

Mycobiology.2022;50(4):231-237. doi:10.1080/12298093.2022.2116816

Penicillium species have been actively studied in various fields, and many new and unrecorded species continue to be reported in Korea. Moreover, unidentified and misidentified Korean Penicillium species still exist in GenBank. Therefore, it is necessary to revise the Korean Penicillium inventory based on accurate identification. We collected Korean Penicillium nucleotide sequence records from GenBank using the newly developed software, GenMine, and re-identified Korean Penicillium based on the maximum likelihood trees. A total of 1681 Korean Penicillium GenBank nucleotide sequence records were collected from GenBank. In these records, 1208 strains with four major genes (Internal Transcribed Spacer rDNA region, b-tubulin, Calmodulin and RNA polymerase II) were selected for Penicillium reidentification. Among 1208 strains, 927 were identified, 82 were identified as other genera, the rest remained undetermined due to low phylogenetic resolution. Identified strains consisted of 206 Penicillium species, including 156 recorded species and 50 new species candidates. However, 37 species recorded in the national list of species in Korea were not found in GenBank. Further studies on the presence or absence of these species are required through literature investigation, additional sampling, and sequencing. Our study can be the basis for updating the Korean Penicillium inventory.

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Seven Undescribed Aspergillus Species from Different Niches in Korea

Monmi PANGGING ; Thuong T. T. NGUYEN ; Hyang Burm LEE

Mycobiology.2022;50(4):189-202. doi:10.1080/12298093.2022.2116158

An investigation of species of the genus Aspergillus present in arthropod, freshwater, and soil led to the discovery of seven undescribed species in Korea. Based on their morphological characteristics and molecular phylogeny analyses using a combined data set of b-tubulin (BenA) and calmodulin (CaM) sequences, the isolated strains CNUFC IGS2-5, CNUFC YJ1-19, CNUFC WD27, CNUFC U8-70, CNUFC AS2-24, CNUFC S32-1, and CNUFC U7-48, were identified as Aspergillus brunneoviolaceus, A. capensis, A. floccosus, A. inflatus, A. parvulus, A.polyporicola, and A. spelaeus, respectively. In the present study, the detailed morphological descriptions and phylogenetic relationships of these species are provided.

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Effect of Temperature, pH, and Media on the Mycelial Growth ofTuber koreanum

Ju-Hui GWON ; Hyeok PARK ; Ahn-Heum EOM

Mycobiology.2022;50(4):238-243. doi:10.1080/12298093.2022.2112586

Members of the genus Tuber are ectomycorrhizal fungi; this genus includes more than 180 species worldwide. In the present study, the optimal pH, temperature, and medium suitable for the mycelial growth of the Korean truffle, Tuber koreanum, were determined. Mycelium of T. koreanum, isolated from fruiting bodies collected in Korea, was used to investigate the effects of these environmental factors. The results showed that malt extract agar and potato dextrose agar were the most suitable for the mycelial growth of T. koreanum when cultured at a pH of 6.0 at 25 C for 30 days.

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