Proteomic Approach To The Differentially Expressed Proteins Between Bursaphelenchus Xylophilus And B.mucronatus

Pine wilt disease is one of the most important pine tree diseases, and its causal agent is Bursaphelenchus xylophilus, which is a great threat to the pine forestry in many countries such as China Japan, Korea, the US, Canada, Mexico, and Nigeria, etc. It is one of the most important quarantine organisms, and also listed on A2 in China, which lead to imponderable loss every year. However, it is difficult to differentiate isolates of B. xylophilus from the closely related species Bursaphelenchus mucronatus, which is not pathogenic to pine trees, especially the juveniles. It makes the quarantine and prevention very difficult to carry on. Presently, technologies of detecting B. xylophilus are merely limited in traditional morphological, serology and PCR molecular identification. Research in mechanism of pathology is just limited in tissue pathology. However, few researches in differential proteomics have been reported. Therefore, it is of great significantly to investigate exact identification and pathological mechanism of B. xylophilus by utilizing method of differential proteomics.In this study, we extracted the total proteins of the two nematodes, and applied two-dimensional electrophoresis and mass spectrometry to study the proteomics of the two nematodes. Through analyzing differential proteomics of the two nematodes, we found an approach to identify the two nematodes exactly. Furthermore, an advanced research of this will uncover the pathological mechanism of B. xylophilus.In equal conditions, we extracted the total proteins of the two nematodes, and applied two-dimensional electrophoresis to analyze the differences between the two nematodes. As a result, we have obtained 50 differentiate proteins,25 of which were from B. xylophilus and others were from B. mucronatus.45 proteins were determined by matrix-assisted laser desorption/ ionization-time of flight mass spectrometry (MALDI-TOF-MS),and identified protein database matching (PMF),22 of which were from B. xylophilus and others were from B. mucronatus. Further analysis of these 45 proteins showed them involved in differentiate metabolism pathway and signal transduction, including metabolism associated proteins, signal pathway proteins and protein synthesis associated proteins. It is supposed that these different proteins participate in different metabolism pathways and signal transduction pathways, and these discriminations are related to B. xylophilus'capability to infect while B. mucronatus'low capability to infect. Most of these different proteins are protein enzymes, which provide evidences to"the theory of enzyme"which consider that it is enzymes from B. xylophilus cause atrophy of cell wall and cell membrane from parenchyma cells.It is shown that, there are something different in metabolism pathways of carbohydrate and signal transductions pathways between the two nematodes. This discovering support the point of view that B. xylophilus is the exclusive pathogeny of pine wilt disease, and"the theory of enzyme".Identifying the differentiate proteins between the two nematodes will enable us to identify the two nematodes rapidly and accurately. Integrating the differentiate proteins and other data between the two nematodes will assist us to reveal the virulence of B. xylophilus and search for proteins and genes which is related to pine wilt disease.