Monochamus Alternatus Attractant And Molecular Detection Technique Of Bursaphelenchus Xylophilus

Pine Wilt Disease is a kind of pine destructive disease which is severely threatening forests safety both in our country and worldwide. Its causal agent, Bursaphelenchus xylophilus(Steiner & Buhrer)Nickle, has been placed into the class of quarantine pests in many countries because of its causing severe damages and being hard to prevent and manage.The pine wood nematode is mainly vectored by Monochamus alternatus, which are not only the vector of Bursaphelenchus xylophilus but also the important stem borers of native coniferous forests. These beetles transmit the pine wood nematode to new host trees through feeding and ovipositing. So controlling the population of Monochamus alternatus could not only relieve direct damages but prevent Bursaphelenchus xylophilus spreading.In addition, there area little discrepancy between Bursaphelenchus xylophilus and its valid relative species such as B. fraudulentus, B. mucronatus, B. abruptus, B. conicaudatus, B. luxuriosae, B. doui, B. curvicaudatus and so on, and it is very hard to discriminate their larvae. Thus, constructing a set of Bursaphelenchus xylophilus fast diagnostic method is very important both to port quarantine and keeping forest health of our country.For two key steps of preventing and controlling B. xylophilus, Monochamus alternatus attractant and identification B. xylophilus, four research works have been studied in this paper. The first work was the study on volatiles constituents of Pinus massoniana, P. thunbergii, P. teada, P. elliottii and Cedrus deodara, which are all host plants of Monochamus alternatus. The second thing was Electroantennogram(EAG) and behavioural responses of Monochamus alternatus in different developmental stages to extracts, monomer compound, and combination compound. The third study was about field testing of attacting Monochamus alternatus with concoct attractant .The last work was to design TaqMan-MGB probe for quick detection of B. xylophilus. The mainly contents and results of study were highlighted following: Conducting identification of the volatiles compounds extracted from branch and needle of five host plants by Solid Phase Microextraction and Steam Distillation;Studying on comparison of Chemical composition of volatiles between healthy and infested branches of host plant; Studying on response of EAG and Olfactory of adult M. alternatus to extracts of healthy and infested pine trees;Studying on response of EAG and Olfactory of adult M. alternatus to mainly volatile compounds of the host pine trees;Studying on the selected attractants and evaluation of the traps for the capture of M. alternatus in forest field;Comparative study on detection tecniques for B. xylophilus with specific premer pairs polymerase chain reaction and ITS-PCR-PFLP; Conducting study on real-time PCR detection technique and invented a quick real-time detection experiment box;Conducting identification of B. xylophilus collected from different places by using the quick real-time PCR detection technique and This probe (TaqMan-MGB) and primers(F/R) have been applied for nation patent protection (200510026710.2).