High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity

The ecology of total fungal communities in grapevine is so far largely derived from studies on culture-dependent methods or cultivation-independent rDNA approaches. Sequencing the ribosomal RNA transcripts (rRNA) would rather reveal the functionally and metabolically active important taxa of the fungal community and provide insights into its activity in the wood. The present study investigated changes in the potentially active fungal communities of internal grapevine wood after Hot-Water Treatment (HWT) in planting material from Czech Republic and Spain at two different times during the propagation process and from two plant zones. We examined fungal communities using both traditional isolation into culture and high-throughput amplicon sequencing (HTAS) of the internal transcribed spacer 2 (ITS2) region in extracted total RNA. HTAS from metatranscriptomic RNA increased the resolution of the fungal community analysis and revealed a highly diverse mycobiota of grapevine wood compared to the traditional method. Fungal diversity differed between grapevine genotypes and showed a temporal variation over the vegetative period. Grapevine planting materials exhibited high fungal diversity after HWT, which demonstrates that the HWT process does not sterilize the internal wood of grapevine. HWT reduced the infection caused by fungal trunk disease pathogens but was not completely effective in eliminating their growth. This study provides important and practically useful insights into the dynamics of active fungal communities in hot-water treated plants, and represents the first study of active fungal communities on grapevine grafted plants by comparing traditional and next-generation sequencing methods.

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Main Authors: Eichmeier, Ales, Pečenka, Jakub, Peňázová, Eliska, Baránek, Miroslav, Català-García, Santiago, León, Maela, Armengol, Josep, Gramaje, David
Other Authors: Ministry of Education, Youth and Sports (Czech Republic)
Format: artículo biblioteca
Language:English
Published: Elsevier 2018-12
Subjects:Endophytic fungi, Grapevine trunk diseases, ITS2, Metatranscriptomics, Mycobiota, NGS, Plant microbiome,
Online Access:http://hdl.handle.net/10261/192891
http://dx.doi.org/10.13039/501100000921
http://dx.doi.org/10.13039/100007652
http://dx.doi.org/10.13039/501100000780
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spelling dig-icvv-es-10261-1928912019-11-06T08:18:09Z High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity Eichmeier, Ales Pečenka, Jakub Peňázová, Eliska Baránek, Miroslav Català-García, Santiago León, Maela Armengol, Josep Gramaje, David Ministry of Education, Youth and Sports (Czech Republic) CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) European Commission European Cooperation in Science and Technology Pečenka, Jakub [0000-0001-6195-7592] Armengol, Josep [0000-0003-3815-8578] Endophytic fungi Grapevine trunk diseases ITS2 Metatranscriptomics Mycobiota NGS Plant microbiome The ecology of total fungal communities in grapevine is so far largely derived from studies on culture-dependent methods or cultivation-independent rDNA approaches. Sequencing the ribosomal RNA transcripts (rRNA) would rather reveal the functionally and metabolically active important taxa of the fungal community and provide insights into its activity in the wood. The present study investigated changes in the potentially active fungal communities of internal grapevine wood after Hot-Water Treatment (HWT) in planting material from Czech Republic and Spain at two different times during the propagation process and from two plant zones. We examined fungal communities using both traditional isolation into culture and high-throughput amplicon sequencing (HTAS) of the internal transcribed spacer 2 (ITS2) region in extracted total RNA. HTAS from metatranscriptomic RNA increased the resolution of the fungal community analysis and revealed a highly diverse mycobiota of grapevine wood compared to the traditional method. Fungal diversity differed between grapevine genotypes and showed a temporal variation over the vegetative period. Grapevine planting materials exhibited high fungal diversity after HWT, which demonstrates that the HWT process does not sterilize the internal wood of grapevine. HWT reduced the infection caused by fungal trunk disease pathogens but was not completely effective in eliminating their growth. This study provides important and practically useful insights into the dynamics of active fungal communities in hot-water treated plants, and represents the first study of active fungal communities on grapevine grafted plants by comparing traditional and next-generation sequencing methods. This work was supported by Projects No. LD14051 from the Ministry of Education, Youth and Sports, Czech Republic covered by COST Action FA1303, and No. CZ.02.1.01/0.0/0.0/16_025/0007314 “Multidisciplinary research to increase application potential of nanomaterials in agricultural practice” from EFRR (Grant provider: Ministerstvo školství, mládeže a tělovýchovy České republiky (MŠMT). Access to computing and storage facilities owned by parties and projects contributing to the National Grid Infrastructure MetaCentrum provided under the programme "Projects of Large Research, Development, and Innovations Infrastructures" (CESNET LM2015042), is greatly appreciated. David Gramaje was supported by the DOC-INIA program from the National Institute for Agronomic Research (INIA), co-funded by the European Social Fund. The authors are grateful for the technical assistance of Jesús Alfonso Crespo and Aitor Gavara Vidal. Peer reviewed 2019-10-18T10:19:14Z 2019-10-18T10:19:14Z 2018-12 artículo http://purl.org/coar/resource_type/c_6501 Fungal Ecology 36: 26-38 (2018) 1754-5048 http://hdl.handle.net/10261/192891 10.1016/j.funeco.2018.07.011 http://dx.doi.org/10.13039/501100000921 http://dx.doi.org/10.13039/100007652 http://dx.doi.org/10.13039/501100000780 en http://dx.doi.org/10.1016/j.funeco.2018.07.011 Sí none Elsevier
institution ICVV ES
collection DSpace
country España
countrycode ES
component Bibliográfico
access En linea
databasecode dig-icvv-es
tag biblioteca
region Europa del Sur
libraryname Biblioteca del ICVV España
language English
topic Endophytic fungi
Grapevine trunk diseases
ITS2
Metatranscriptomics
Mycobiota
NGS
Plant microbiome
Endophytic fungi
Grapevine trunk diseases
ITS2
Metatranscriptomics
Mycobiota
NGS
Plant microbiome
spellingShingle Endophytic fungi
Grapevine trunk diseases
ITS2
Metatranscriptomics
Mycobiota
NGS
Plant microbiome
Endophytic fungi
Grapevine trunk diseases
ITS2
Metatranscriptomics
Mycobiota
NGS
Plant microbiome
Eichmeier, Ales
Pečenka, Jakub
Peňázová, Eliska
Baránek, Miroslav
Català-García, Santiago
León, Maela
Armengol, Josep
Gramaje, David
High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
description The ecology of total fungal communities in grapevine is so far largely derived from studies on culture-dependent methods or cultivation-independent rDNA approaches. Sequencing the ribosomal RNA transcripts (rRNA) would rather reveal the functionally and metabolically active important taxa of the fungal community and provide insights into its activity in the wood. The present study investigated changes in the potentially active fungal communities of internal grapevine wood after Hot-Water Treatment (HWT) in planting material from Czech Republic and Spain at two different times during the propagation process and from two plant zones. We examined fungal communities using both traditional isolation into culture and high-throughput amplicon sequencing (HTAS) of the internal transcribed spacer 2 (ITS2) region in extracted total RNA. HTAS from metatranscriptomic RNA increased the resolution of the fungal community analysis and revealed a highly diverse mycobiota of grapevine wood compared to the traditional method. Fungal diversity differed between grapevine genotypes and showed a temporal variation over the vegetative period. Grapevine planting materials exhibited high fungal diversity after HWT, which demonstrates that the HWT process does not sterilize the internal wood of grapevine. HWT reduced the infection caused by fungal trunk disease pathogens but was not completely effective in eliminating their growth. This study provides important and practically useful insights into the dynamics of active fungal communities in hot-water treated plants, and represents the first study of active fungal communities on grapevine grafted plants by comparing traditional and next-generation sequencing methods.
author2 Ministry of Education, Youth and Sports (Czech Republic)
author_facet Ministry of Education, Youth and Sports (Czech Republic)
Eichmeier, Ales
Pečenka, Jakub
Peňázová, Eliska
Baránek, Miroslav
Català-García, Santiago
León, Maela
Armengol, Josep
Gramaje, David
format artículo
topic_facet Endophytic fungi
Grapevine trunk diseases
ITS2
Metatranscriptomics
Mycobiota
NGS
Plant microbiome
author Eichmeier, Ales
Pečenka, Jakub
Peňázová, Eliska
Baránek, Miroslav
Català-García, Santiago
León, Maela
Armengol, Josep
Gramaje, David
author_sort Eichmeier, Ales
title High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
title_short High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
title_full High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
title_fullStr High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
title_full_unstemmed High-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
title_sort high-throughput amplicon sequencing-based analysis of active fungal communities inhabiting grapevine after hot-water treatments reveals unexpectedly high fungal diversity
publisher Elsevier
publishDate 2018-12
url http://hdl.handle.net/10261/192891
http://dx.doi.org/10.13039/501100000921
http://dx.doi.org/10.13039/100007652
http://dx.doi.org/10.13039/501100000780
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