title: Screening and functional analysis of the peroxiredoxin specifically expressed in Bursaphelenchus xylophilus--the causative agent of pine wilt disease.
abstract: The pine wood nematode, Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. Accurately differentiating B. xylophilus from other nematodes species, especially its related species B. mucronatus, is important for pine wood nematode detection. Thus, we attempted to identify a specific protein in the pine wood nematode using proteomics technology. Here, we compared the proteomes of B. xylophilus and B. mucronatus using Two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization -time-of-flight/time-of-flight (MALDI-TOF/TOF-MS) technologies. In total, 15 highly expressed proteins were identified in B. xylophilus compared with B. mucronatus. Subsequently, the specificity of the proteins identified was confirmed by PCR using the genomic DNA of other nematode species. Finally, a gene encoding a specific protein (Bx-Prx) was obtained. This gene was cloned and expressed in E. coli. The in situ hybridisation pattern of Bx-Prx showed that it was expressed strongly in the tail of B. xylophilus. RNAi was used to assess the function of Bx-Prx, the results indicated that the gene was associated with the reproduction and pathogenicity of B. xylophilus. This discovery provides fundamental information for identifying B. xylophilus via a molecular approach. Moreover, the purified recombinant protein has potential as a candidate diagnostic antigen of pine wilt disease, which may lead to a new immunological detection method for the pine wood nematode.
authors.affiliation: College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, Jiangsu, China. fuhanyu2019@gmail.com.
authors.forename: Han-Yu
authors.initials: HY
authors.lastname: Fu
authors.affiliation: College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, Jiangsu, China. renjiahong04@gmail.com.
authors.forename: Jia-Hong
authors.initials: JH
authors.lastname: Ren
authors.affiliation: College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, Jiangsu, China. Lhuang@njfu.edu.cn.
authors.forename: Lin
authors.initials: L
authors.lastname: Huang
authors.affiliation: College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, Jiangsu, China. lihao201404@gmail.com.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.affiliation: College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, Jiangsu, China. jryenjfu2014@gmail.com.
authors.forename: Jian-Ren
authors.initials: JR
authors.lastname: Ye
mesh_terms: D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000378:metabolism; D017403:In Situ Hybridization; D054464:Peroxiredoxins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D028223:Pinus / Q000254:growth & development / Q000469:parasitology; D010935:Plant Diseases / Q000469:parasitology; D020543:Proteome / Q000032:analysis; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D036226:Seedlings / Q000187:drug effects / Q000254:growth & development; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017187:Tylenchida / Q000378:metabolism*
chemical_list: D020543:Proteome; D012330:RNA, Double-Stranded; D011994:Recombinant Proteins; D054464:Peroxiredoxins