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.

A 2-Cys peroxiredoxin in response to oxidative stress in the pine wood nematode, Bursaphelenchus xylophilus

“…Silencing of CePrx2 in C. elegans resulted in a stunted development and a significantly reduced brood size 32 . Similarly, knocking down BxPrx significantly reduced the propagation in B. xylophilus 50 . Our previous study found that anti-serum of BxPrx could only detect protein samples extracted from virulent B. xylophilus but not B. mucronatus 28 , suggesting that BxPrx could be used as a molecular marker to identify the virulent B. xylophilus 50 .…”

“…Similarly, knocking down BxPrx significantly reduced the propagation in B. xylophilus 50 . Our previous study found that anti-serum of BxPrx could only detect protein samples extracted from virulent B. xylophilus but not B. mucronatus 28 , suggesting that BxPrx could be used as a molecular marker to identify the virulent B. xylophilus 50 . A shorter life cycle and higher proliferation rate are the fundamental differences between avirulent and virulent B. xylophilus , therefore, BxPrx could play a role in the pathogenicity of B. xylophilus .…”

“…In conclusion, we suggest that BxPrx is a key genetic factor that facilitates the infestation and distribution of B. xylophilus within pine hosts, and consequently the spread of pine wilt disease. With the advent of functional genomics tools, including RNA interference (RNAi) 51 and transgenic approach 50 , genetic basis governing the transmission and infection of pine wilt disease is a logic choice for future research. A better understanding of how nematodes overcome oxidative stresses generated from their own metabolic processes and from the innate immune responses in pine hosts will provide novel targets for the long-term, sustainable management of this devastating pine disease.…”

EvaluatingEucalyptusleaf colonization byBrasilonema octagenarum(Cyanobacteria, Scytonemataceae) usingin plantaexperiments and genomics

“…Three copies of the twitching motility gene pilT were annotated in the UFV-E1 genome, important for spreading pathogen cells on the surface of the host (Weller-Stuart et al, 2017). Three translated protein sequences from the UFV-E1 genome have shown similarity with catalase-peroxidases and peroxiredoxins from PHI-base (accessions A4QUT2, B0LFQ7 and G4MXC5), enzymes with very important roles in the maintenance of redox homeostasis and the protection of pathogen cells from oxidative damage brought by hydrogen peroxide produced by host cells (Tanabe et al, 2011;Fu et al, 2014;Mir et al, 2015;Santander, Figàs-Segura & Biosca, 2018). Genes for other proteins potentially involved in detoxification have also been predicted, including multidrug efflux pumps (D4HXR8 and Q7WTQ9), which may confer resistance to phytoalexins (Burse, Weingart & Ullrich, 2004), and S-(hydroxymethyl)gluthatione dehydrogenase (G4N4N6), which is produced by phytopathogens as protection against damage from nitric oxide (Zhang et al, 2015b).…”

Identification of Potential Nematicidal Compounds against the Pine Wood Nematode, Bursaphelenchus xylophilus through an In Silico Approach

“…The availability of the completely sequenced genome of B. xylophilus has opened up the possibility of investigating the most prominent proteins that play a crucial role in the survival and parasitism of the nematode as novel drug targets [ 16 , 17 ]. Hence in this study, cathepsin L-like cysteine proteinase that is involved in post embryonic development [ 18 ], 2-cysteine peroxiredoxins that regulates reproduction and pathogenecity [ 19 ], heat shock protein 90 (HSP90) that helps adaptation to different climatic conditions [ 20 ], venom allergen proteins (VAP) that manifests the invasion of parasitic genes [ 21 ] and tubulin that regulates the microtubule, mitosis and motility [ 22 , 23 ] of B. xylophilus are considered as drug targets. In general, the anti-nematode drugs are classified into two main types based on their target sites, such as drugs targeting membrane ion channels and other class of drugs that acts on biochemical targets.…”