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Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization

BACKGROUND: Pine wilt disease is caused by the pine wood nematode, Bursaphelenchus xylophilus, which threatens pine forests and forest ecosystems worldwide and causes serious economic losses. In the 40 years since the pathogen was identified, the physiological changes occurring as the disease progre...

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Autores principales: Hirao, Tomonori, Fukatsu, Eitaro, Watanabe, Atsushi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3398268/
https://www.ncbi.nlm.nih.gov/pubmed/22272988
http://dx.doi.org/10.1186/1471-2229-12-13
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author Hirao, Tomonori
Fukatsu, Eitaro
Watanabe, Atsushi
author_facet Hirao, Tomonori
Fukatsu, Eitaro
Watanabe, Atsushi
author_sort Hirao, Tomonori
collection PubMed
description BACKGROUND: Pine wilt disease is caused by the pine wood nematode, Bursaphelenchus xylophilus, which threatens pine forests and forest ecosystems worldwide and causes serious economic losses. In the 40 years since the pathogen was identified, the physiological changes occurring as the disease progresses have been characterized using anatomical and biochemical methods, and resistant trees have been selected via breeding programs. However, no studies have assessed the molecular genetics, e.g. transcriptional changes, associated with infection-induced physiological changes in resistant or susceptible trees. RESULTS: We constructed seven subtractive suppression hybridization (SSH) cDNA libraries using time-course sampling of trees inoculated with pine wood nematode at 1, 3, or 7 days post-inoculation (dpi) in susceptible trees and at 1, 3, 7, or 14 dpi in resistant trees. A total of 3,299 sequences was obtained from these cDNA libraries, including from 138 to 315 non-redundant sequences in susceptible SSH libraries and from 351 to 435 in resistant SSH libraries. Using Gene Ontology hierarchy, those non-redundant sequences were classified into 15 subcategories of the biological process Gene Ontology category and 17 subcategories of the molecular function category. The transcriptional components revealed by the Gene Ontology classification clearly differed between resistant and susceptible libraries. Some transcripts were discriminative: expression of antimicrobial peptide and putative pathogenesis-related genes (e.g., PR-1b, 2, 3, 4, 5, 6) was much higher in susceptible trees than in resistant trees at every time point, whereas expression of PR-9, PR-10, and cell wall-related genes (e.g., for hydroxyproline-rich glycoprotein precursor and extensin) was higher in resistant trees than in susceptible trees at 7 and 14 dpi. CONCLUSIONS: Following inoculation with pine wood nematode, there were marked differences between resistant and susceptible trees in transcript diversity and the timing and level of transcripts expressed in common; in particular, expression of stress response and defense genes differed. This study provided new insight into the differences in the physiological changes between resistant and susceptible trees that have been observed in anatomical and biochemical studies.
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spelling pubmed-33982682012-07-18 Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization Hirao, Tomonori Fukatsu, Eitaro Watanabe, Atsushi BMC Plant Biol Research Article BACKGROUND: Pine wilt disease is caused by the pine wood nematode, Bursaphelenchus xylophilus, which threatens pine forests and forest ecosystems worldwide and causes serious economic losses. In the 40 years since the pathogen was identified, the physiological changes occurring as the disease progresses have been characterized using anatomical and biochemical methods, and resistant trees have been selected via breeding programs. However, no studies have assessed the molecular genetics, e.g. transcriptional changes, associated with infection-induced physiological changes in resistant or susceptible trees. RESULTS: We constructed seven subtractive suppression hybridization (SSH) cDNA libraries using time-course sampling of trees inoculated with pine wood nematode at 1, 3, or 7 days post-inoculation (dpi) in susceptible trees and at 1, 3, 7, or 14 dpi in resistant trees. A total of 3,299 sequences was obtained from these cDNA libraries, including from 138 to 315 non-redundant sequences in susceptible SSH libraries and from 351 to 435 in resistant SSH libraries. Using Gene Ontology hierarchy, those non-redundant sequences were classified into 15 subcategories of the biological process Gene Ontology category and 17 subcategories of the molecular function category. The transcriptional components revealed by the Gene Ontology classification clearly differed between resistant and susceptible libraries. Some transcripts were discriminative: expression of antimicrobial peptide and putative pathogenesis-related genes (e.g., PR-1b, 2, 3, 4, 5, 6) was much higher in susceptible trees than in resistant trees at every time point, whereas expression of PR-9, PR-10, and cell wall-related genes (e.g., for hydroxyproline-rich glycoprotein precursor and extensin) was higher in resistant trees than in susceptible trees at 7 and 14 dpi. CONCLUSIONS: Following inoculation with pine wood nematode, there were marked differences between resistant and susceptible trees in transcript diversity and the timing and level of transcripts expressed in common; in particular, expression of stress response and defense genes differed. This study provided new insight into the differences in the physiological changes between resistant and susceptible trees that have been observed in anatomical and biochemical studies. BioMed Central 2012-01-24 /pmc/articles/PMC3398268/ /pubmed/22272988 http://dx.doi.org/10.1186/1471-2229-12-13 Text en Copyright ©2012 Hirao et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Hirao, Tomonori
Fukatsu, Eitaro
Watanabe, Atsushi
Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization
title Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization
title_full Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization
title_fullStr Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization
title_full_unstemmed Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization
title_short Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization
title_sort characterization of resistance to pine wood nematode infection in pinus thunbergii using suppression subtractive hybridization
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3398268/
https://www.ncbi.nlm.nih.gov/pubmed/22272988
http://dx.doi.org/10.1186/1471-2229-12-13
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