Sino-America Joint Study On Genetic Varieties And PCR Identification Of Anoplophora Glabripennis

Anoplophora glabripernnis (Coleoptera/Cerambycidae). or Asian longhorned beetle(ALB). As a main pest in the forest of our country, endangers broad leaf trees, such as poplar, willow and maple, etc. mainly. The damage is very serious in our country's North China, the Northwest and the Northeast ( so called" three northern area of China") area, that has become a great obstacles of improving the ecological environment by planting poplar in North China. According to estimating, the loss caused by Anoplophora glabripernnis and other poplar beetles in our country reaches 2 billion yuan every year.Since 1996, ALB infestation has been discovered in New York, Chicago and New Jersey in U.S.A. in succession. The discovery of this beetle has caused great attention of U.S.A. public and government. American side statistics show, as soon as this kind of beetle spreads in U.S.A., 70 million hectares of city afforestation woods, the sugar refining industries and tourism will be threaten directly with the direct economic loss reach 138 billionth dollars (1999 Yulin, Chen Hong 2000). U.S.A. thought that this pest imported from China, and signed an interim rule that aimed at imported wooden packing materials from China on September 11, 1998. The statistics show, this decree influences 1/3 to 1/2 of our country's commodities export to America directly, and bring enormous loss to our country's foreign trade. To insure the normal progression of Sino-USA trade, Chinese side issued Document No.104 [National quarantine 1998], call for strengthening the inspection and quarantine of the wood packing materials of goods export to USA.The main result as follows:1 RAPD and mtDNA sequencing protocols on ALB have been established, results obtained by different approach are consistent, showed the technology adopted is reliableThis study investigated the genetic diversity of Anoplophora glabripernnis using Random Amplified Polymorphic DNA (RAPD) assay and DNA sequencing techniques. Outcomes of three RAPD experiments carried out in different period using different specimen and different set of random primers are consistent, thus prove that RAPD method is stable, sensitive, reliable. On the other hand, the DNA sequencing results of mtDNA COI gene give better expression to the genetic relationships of ALB populations, Unanimous basically with the result of RAPD, indicated that ALB mtDNA COI gene sequence is suited to analyze the genetic relation among the beetle populations. The application of two kinds of technology gives the same result, indicate that the conclusion draw from our study is objective. The sample source gathered in research is wide, representative is strong.2 Study on mtDNA sequence and its genetic characters of Anoplophora glabripennisSeven mitochondriaL DNA regions of Anoplophora glabripennis and its sibling species are sequenced to understand the molecular characteristics of mtDNA. The results show that the base variation of the seven mtDNA gene sequences of Anoplcphora glabripennis and those of its sibling species is above 11.3%, whereas the base variation among Anoplophora glabripennis species group and its individuals is less than 3%, nearly no variation existed in most genes, these character imply that these mitochondrial genes are conserved intraspeciesly and could be used in identification and phylogenetic analysis of Anoplophora glabripennis. Our study also suggest that Anoplophora glabripennis and A.nobilis might belong to same species as their sequence variation in these genes are negligible. In the sequencing of CO I Cytb genes, sequences with significant variation are found in some individuals ,sometimes they are similar to those of the sibling. It remains to be elucidated that weather this is due to the presence of nuclear pseudogene, or it may arise frommitochondrial inleakage.3 Development of the molecular biology identification protocol and PCR diagnosis reagent kit of A glabripennisSpecimens of five usual Anoplophora species and 2 foreign species were used in this study, in which 12 A.glabripermis populations a...