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Development of an efficient PCR-based diagnosis protocol for the identification of the pinewood nematode, Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

Development of an efficient PCR-based diagnosis protocol for the identification of the... AbstractPine wood wilt disease caused by the pine wood nematode, Bursaphelenchusxylophilus , has been a serious problem in the southern regions of Korea.Efficient diagnosis of B. xylophilus from infected pine wood specimens iscritical for the management of this pest. Traditional microscopicexamination often results in an erroneous identification because a closelyrelated non-pathogenic species, B. mucronatus, has a great degree ofmorphological similarity to B. xylophilus. In an attempt to search forreliable molecular markers for the discrimination of these species, we havecloned the 5S rRNA genomic DNA fragments containing both coding andintergenic spacer (IGS) regions from B. xylophilus and B. mucronatus througha homology-probing PCR strategy. Sequence analyses revealed that codingsequences of the 5S rRNA gene from the two species are almost identical(98.3% homology) but that the IGS sequences differ substantially between thespecies. Based on the IGS sequence differences (69.7% homology), we designedspecies-specific primer sets and developed a PCR-based diagnosis protocolfor the identification and discrimination of the two nematode species on amolecular basis. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nematology Brill

Development of an efficient PCR-based diagnosis protocol for the identification of the pinewood nematode, Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

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Publisher
Brill
Copyright
Copyright © Koninklijke Brill NV, Leiden, The Netherlands
ISSN
1388-5545
eISSN
1568-5411
DOI
10.1163/1568541041217915
Publisher site
See Article on Publisher Site

Abstract

AbstractPine wood wilt disease caused by the pine wood nematode, Bursaphelenchusxylophilus , has been a serious problem in the southern regions of Korea.Efficient diagnosis of B. xylophilus from infected pine wood specimens iscritical for the management of this pest. Traditional microscopicexamination often results in an erroneous identification because a closelyrelated non-pathogenic species, B. mucronatus, has a great degree ofmorphological similarity to B. xylophilus. In an attempt to search forreliable molecular markers for the discrimination of these species, we havecloned the 5S rRNA genomic DNA fragments containing both coding andintergenic spacer (IGS) regions from B. xylophilus and B. mucronatus througha homology-probing PCR strategy. Sequence analyses revealed that codingsequences of the 5S rRNA gene from the two species are almost identical(98.3% homology) but that the IGS sequences differ substantially between thespecies. Based on the IGS sequence differences (69.7% homology), we designedspecies-specific primer sets and developed a PCR-based diagnosis protocolfor the identification and discrimination of the two nematode species on amolecular basis.

Journal

NematologyBrill

Published: Jan 1, 2004

Keywords: PINE WOOD WILT DISEASE; SPECIES-SPECIFIC; PINE WOOD NEMATODE; BURSAPHELECHUS MUCRONATUS; DIAGNOSTICS; IGS; 5S RRNA; PCR

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