Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR
<i>Serpula himantioides</i> is a wood-decaying fungal pathogen which is widespread and causes potentially serious butt rot in living trees of numerous coniferous plantation species. This study aimed to quantify <i>S. himantioides</i> in the wood of Sawara cypress (<i>Ch...
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MDPI AG
2022-09-01
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| Series: | Forests |
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| Online Access: | https://www.mdpi.com/1999-4907/13/9/1429 |
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| author | Ryusei Haraguchi Toshihide Hirao Toshihiro Yamada |
| author_facet | Ryusei Haraguchi Toshihide Hirao Toshihiro Yamada |
| author_sort | Ryusei Haraguchi |
| collection | DOAJ |
| description | <i>Serpula himantioides</i> is a wood-decaying fungal pathogen which is widespread and causes potentially serious butt rot in living trees of numerous coniferous plantation species. This study aimed to quantify <i>S. himantioides</i> in the wood of Sawara cypress (<i>Chamaecyparis pisifera</i>) with butt rot using real-time polymerase chain reaction (real-time PCR). Species–specific primers were designed for the internal transcribed spacer 2 (ITS2) regions of ribosomal DNA (rDNA) of <i>S. himantioides</i>. The specificity of the designed primer set was tested by end-point PCR and amplicon sequencing. End-point PCR assays were positive for <i>S. himantioides</i> and negative for <i>S. lacrymans,</i> which belongs to the same genus as <i>S. himantioides</i>, <i>Coniophora puteana</i> of the same family as <i>S. himantioides</i>, and other wood decay fungi. Taxonomy assignment based on amplicon sequencing detected only <i>Serpula</i> spp., and most of them were identified as <i>S. himantioides</i>. The linearity of the calibration curve for absolute quantification by real-time PCR assays was confirmed in the range from 10<sup>1</sup> to 10<sup>8</sup> copies. This molecular assay method using real-time PCR could detect trace amounts of <i>S. himantioides</i> in decayed wood, showing the applicability for early diagnostics of butt rot emergence in forests. |
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| issn | 1999-4907 |
| language | English |
| last_indexed | 2024-03-10T00:00:34Z |
| publishDate | 2022-09-01 |
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| series | Forests |
| spelling | doaj.art-9df8d0a593f14c7f980fc5915fb2612e2023-11-23T16:17:18ZengMDPI AGForests1999-49072022-09-01139142910.3390/f13091429Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCRRyusei Haraguchi0Toshihide Hirao1Toshihiro Yamada2The University of Tokyo Chichibu Forest, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-49 Hinoda-machi, Chichibu, Saitama 368-0034, JapanThe University of Tokyo Chichibu Forest, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-49 Hinoda-machi, Chichibu, Saitama 368-0034, JapanThe University of Tokyo Chichibu Forest, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-49 Hinoda-machi, Chichibu, Saitama 368-0034, Japan<i>Serpula himantioides</i> is a wood-decaying fungal pathogen which is widespread and causes potentially serious butt rot in living trees of numerous coniferous plantation species. This study aimed to quantify <i>S. himantioides</i> in the wood of Sawara cypress (<i>Chamaecyparis pisifera</i>) with butt rot using real-time polymerase chain reaction (real-time PCR). Species–specific primers were designed for the internal transcribed spacer 2 (ITS2) regions of ribosomal DNA (rDNA) of <i>S. himantioides</i>. The specificity of the designed primer set was tested by end-point PCR and amplicon sequencing. End-point PCR assays were positive for <i>S. himantioides</i> and negative for <i>S. lacrymans,</i> which belongs to the same genus as <i>S. himantioides</i>, <i>Coniophora puteana</i> of the same family as <i>S. himantioides</i>, and other wood decay fungi. Taxonomy assignment based on amplicon sequencing detected only <i>Serpula</i> spp., and most of them were identified as <i>S. himantioides</i>. The linearity of the calibration curve for absolute quantification by real-time PCR assays was confirmed in the range from 10<sup>1</sup> to 10<sup>8</sup> copies. This molecular assay method using real-time PCR could detect trace amounts of <i>S. himantioides</i> in decayed wood, showing the applicability for early diagnostics of butt rot emergence in forests.https://www.mdpi.com/1999-4907/13/9/1429<i>Serpula himantioides</i>fungiearly diagnosticsenvironmental samplereal-time PCRamplicon sequencing |
| spellingShingle | Ryusei Haraguchi Toshihide Hirao Toshihiro Yamada Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR Forests <i>Serpula himantioides</i> fungi early diagnostics environmental sample real-time PCR amplicon sequencing |
| title | Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR |
| title_full | Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR |
| title_fullStr | Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR |
| title_full_unstemmed | Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR |
| title_short | Detection and Quantification of <i>Serpula himantioides</i> in the Wood of <i>Chamaecyparis pisifera</i> Butt Rot Trees by Real-Time PCR |
| title_sort | detection and quantification of i serpula himantioides i in the wood of i chamaecyparis pisifera i butt rot trees by real time pcr |
| topic | <i>Serpula himantioides</i> fungi early diagnostics environmental sample real-time PCR amplicon sequencing |
| url | https://www.mdpi.com/1999-4907/13/9/1429 |
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