The wood decay fungi are a diverse taxonomic group that plays a pivotal role in forest carbon cycling. Wood decay fungi use various enzymatic pathways to digest dead or living wood in order to obtain carbon and other nutrients and these enzymatic systems have been exploited for both industrial and m...
The wood decay fungi are a diverse taxonomic group that plays a pivotal role in forest carbon cycling. Wood decay fungi use various enzymatic pathways to digest dead or living wood in order to obtain carbon and other nutrients and these enzymatic systems have been exploited for both industrial and medical applications. Over 600 wood decay fungi species have been described in Korea; however, the recent application of molecular markers has dramatically altered the taxonomy of many of these wood decay fungi at both the genus and species levels. By combining molecular methods, specifically sequences of the internal transcribed spacer region, with traditional morphological characters, this study identified five new species records for Korea in five genera: Aurantiporus, Favolus, Neofavolus, Loweomyces, and Hymenochaetopsis. Three of these genera (Aurantiporus, Favolus, and Loweomyces) were previously unknown in Korea. The relatively simple morphology of the wood decay fungi often leads to ambiguous taxonomic assignment. Therefore, molecular markers are a necessary component of any taxonomic or evolutionary study of wood decay fungi. Our study highlights the need for a more robust and multifaceted approach in investigating new wood decay fungi in Korea.
The wood decay fungi are a diverse taxonomic group that plays a pivotal role in forest carbon cycling. Wood decay fungi use various enzymatic pathways to digest dead or living wood in order to obtain carbon and other nutrients and these enzymatic systems have been exploited for both industrial and medical applications. Over 600 wood decay fungi species have been described in Korea; however, the recent application of molecular markers has dramatically altered the taxonomy of many of these wood decay fungi at both the genus and species levels. By combining molecular methods, specifically sequences of the internal transcribed spacer region, with traditional morphological characters, this study identified five new species records for Korea in five genera: Aurantiporus, Favolus, Neofavolus, Loweomyces, and Hymenochaetopsis. Three of these genera (Aurantiporus, Favolus, and Loweomyces) were previously unknown in Korea. The relatively simple morphology of the wood decay fungi often leads to ambiguous taxonomic assignment. Therefore, molecular markers are a necessary component of any taxonomic or evolutionary study of wood decay fungi. Our study highlights the need for a more robust and multifaceted approach in investigating new wood decay fungi in Korea.
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제안 방법
PCR products were purified using the Expin PCR Purification Kit (GeneAll Biotechnology, Seoul, Korea) according to the manufacturer’s instructions. DNA sequencing was performed by Macrogen (Seoul, Korea), using an ABI3700 automated DNA sequencer.
The specimens were putatively identified based on fruiting body morphology and microscopic characteristics using Nikon 80i light microscope (Nikon, Tokyo, Japan) [11, 12, 18, 30]. Dried tissue was rehydrated in 3% (w/v) KOH and stained in 1% (w/v) phloxine in order to measure key features, and drawings of these macromorphological and micromorphological features were generated. Our primary focus was the measurement of basidia (30 per sample) and basidiospores (10 per sample).
In this study, we combined macro- and microscopic morphological observations with molecular analysis of ITS sequences, and accurately identify five species (in five genera) of wood decay fungi that are new records to Korea. While BLAST searches using ITS sequences can provide valuable preliminary identification of species, incorrectly identified sequences in GenBank is an open database are not uncommon (approximately 20%) [40].
대상 데이터
Eighteen specimens were identified to five species in five genera: Aurantiporus, Favolus, Neofavolus, Loweomyces, and Hymenochaetopsis. A NJ tree was constructed for each genus, with specimens from this study highlighted in gray boxes (Figs. 1~5). One specimen (SFC20140626-03) was identified as Aurantiporus based on ITS sequences and a phylogram was constructed using 23 closely related species with Rigidoporus ulmarius as an outgroup.
Material examined: Korea, Chungcheongnam-do, Boryeong-si, Mt. Seongju, on oak, 25 Jul 2012, Y. W. Lim (SFC20120725-05); Korea, Gyeonggi-do, Gwacheon-si, Mt. Cheonggye, on Actinidia arguta, 7 Jul 2015, Y. W. Lim (SFC20150707-73); Korea, Incheon-si, Ganghwa-gun, Mt. Mani, on hardwood, 18 Aug 2015, Y. W. Lim (SFC20150818-25).
Material examined: Korea, Gyeonggi-do, Guri-si, Dongguneung, on oak, 26 Jun 2014, Y. W. Lim (SFC20140626-03).
Material examined: Korea, Jeju-do, Seogwipo-si, Andeok-ri, on hardwood, 2 Jul 2014, Y. W. Lim (SFC20140702-07); Korea, Jeju-do, Jeju-si, Pyeongdae-ri, on hardwood, 30 Jun 2015, Y. W. Lim (SFC20150630-50).
Six specimens (SFC20130917-07, SFC20140725-33, SFC20140818-10, SFC20140921-24, SFC20150828-34, and SFC20150902-80) clustered into a monophyletic group with the reference sequences of Loweomyces fractipes (bootstrap support, 100%; sequence similarity, 97.8~100.0%) (Fig. 4).
Specimens of wood decay fungi were collected from various locations in Korea between 2012 and 2015. Specimens were dried and deposited in the Seoul National University Fungus Collection (SFC). The specimens were putatively identified based on fruiting body morphology and microscopic characteristics using Nikon 80i light microscope (Nikon, Tokyo, Japan) [11, 12, 18, 30].
이론/모형
Through this project, we discovered five new records of wood rot fungi from Korea. The specimens were identified to the species level using a two-step approach: phylogenetic analysis based on the ITS region and verification using morphological characters. In this paper, we document and describe these new species records using this two-step approach.
성능/효과
Phylogenetic analyses. Eighteen specimens were identified to five species in five genera: Aurantiporus, Favolus, Neofavolus, Loweomyces, and Hymenochaetopsis. A NJ tree was constructed for each genus, with specimens from this study highlighted in gray boxes (Figs.
Five Hymenochaete-like specimens (SFC20140314-10, SFC20140411-08, SFC20140411-20, SFC20140703-24, and SFC20140723-16) formed a monophyletic clade with the reference sequences of Hymenochaetopsis rigidula. The similarity of all the sequences in this clade was high (99.
He & Jiao Yang was recently suggested to be synonymous with Pseudochaete [19]. Four species were recorded previously in Korea as belonging to the genus Hymenochaete: H. corrugata, H. intricata, H. tabacina, and H. yasudae. Through ITS sequence analysis, this study identified the species H.
Four specimens (SFC20120725-05, SFC20150707-73, SFC20150818-25, and SFC20150820-03) formed a monophyletic group with the reference sequences of Favolus acervatus with 100% bootstrap support and their sequence similarity ranged from 99.8% to 100.0% (Fig. 2). Two specimens (SFC20140702-07 and SFC20150630-50) grouped with the reference sequence of Neofavolus mikawai (bootstrap support, 100%; sequence similarity, 99.
0%) and the clade was well supported (100% bootstrap value). Hymenochaetopsis tabacinoides and H. lamellata were the sister species although bootstrap support value was low (Fig. 5).
1~5). One specimen (SFC20140626-03) was identified as Aurantiporus based on ITS sequences and a phylogram was constructed using 23 closely related species with Rigidoporus ulmarius as an outgroup. This specimen formed a monophyletic clade with the reference sequences of Aurantiporus fissilis (bootstrap support, 100%; sequence similarity, 99.
One specimen (SFC20140626-03) was identified as Aurantiporus based on ITS sequences and a phylogram was constructed using 23 closely related species with Rigidoporus ulmarius as an outgroup. This specimen formed a monophyletic clade with the reference sequences of Aurantiporus fissilis (bootstrap support, 100%; sequence similarity, 99.7~100.0%). A.
yasudae. Through ITS sequence analysis, this study identified the species H. rigidula as a new record in Korea. H.
2). Two specimens (SFC20140702-07 and SFC20150630-50) grouped with the reference sequence of Neofavolus mikawai (bootstrap support, 100%; sequence similarity, 99.6~100.0%). While N.
0%). While N. mikawai exhibited strong sequence similarity with both Neofavolus alveolaris (AB735967 and AB735968, 90.0~90.8%) and Neofavolus cremeoalbidus (AB735979 and AB735980, 90.5~91.1%), which are sister species, the evolutionary relationships of these three species are unclear as the cluster was not supported (Fig. 3).
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