Construction and characterization of subtractive stage-specific expressed sequence tag (EST) libraries of the pinewood nematode Bursaphelenchus xylophilus

To establish expressed sequence tag databases of the two life stages (the dispersal and propagative stages) of pinewood nematode Bursaphelenchus xylophilus, subtractive EST libraries that were specific to the dispersal 4th larval stage (D4S) and the pine-grown propagative mixed (PGPS) stage were con...

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Bibliographic Details
Published in:Genomics (San Diego, Calif.) Calif.), 2009-07, Vol.94 (1), p.70-77
Main Authors: Kang, Jae Soon, Lee, Hyoungseok, Moon, Il Sung, Lee, Yi, Koh, Young Ho, Je, Yeon Ho, Lim, Kook-Jin, Lee, Si Hyeock
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Bursaphelenchus xylophilus
Computational Biology - methods
Contig Mapping
Dispersal 4th larva stage
Diverse techniques
ESTs
Expressed Sequence Tags
Fundamental and applied biological sciences. Psychology
Gene Library
Gene ontology
Genes. Genome
Genetics of eukaryotes. Biological and molecular evolution
Life Cycle Stages - genetics
Molecular and cellular biology
Molecular genetics
Nematoda
Nematoda - genetics
Pine-grown propagative mixed stage
Software
Suppressed subtractive hybridization
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Summary:To establish expressed sequence tag databases of the two life stages (the dispersal and propagative stages) of pinewood nematode Bursaphelenchus xylophilus, subtractive EST libraries that were specific to the dispersal 4th larval stage (D4S) and the pine-grown propagative mixed (PGPS) stage were constructed by suppressed subtractive hybridization, and annotated by BLASTx and Gene Ontology (GO). A total of 1112 (57.7%) contigs from the D4S-cDNA library and 1215 (46.7%) contigs from the PGPS-specific cDNA libraries had matched BLASTx hits ( E ≤ 10 − 2 ), among which 913 (47.4%) and 960 (36.9%) contigs, respectively, were classified into three GO categories. A total of 14 genes were selected on the basis of stage-specific abundances and GO subcategories, and their transcription levels were analyzed by quantitative real-time PCR. We discussed the potentials of some stage-specific genes, such as sorbitol dehydrogenase, cysteine protease, venom allergen-like protein, and FMRFamide-like peptide, as diagnosis markers and novel control targets.
ISSN:0888-7543
1089-8646
DOI:10.1016/j.ygeno.2009.03.001