Genetic Difference Of MtDNA COⅠ Gene Of A.glabripennis And Its Sibling Species And PCR Identification Kit Of A.glabripennis

Anoplophora glabripennis (Motschulsky), which widely distribute in China, has been a great economic pest of forest trunk. Since 1996, A.glabripennis infestation has been discovered in New York City, Chicago City and New Jersey in U.S.A. in succession. U.S.A. government thought that this pest imported from China. So in 1998, U.S.A.. government implemented a series of more stern quarantine and inspection regulations to SWPM from China, that had influenced seriously on the export trade of our country and brought the enormous economic losses. As the importance of A.glabripennis to U.S.A. and China, the subject "Genetic differences and identification technology of A.glabripennis "was decided to study by co-operative of both sides after negotiations. This study is part of this subject.A 504bp fragment of mtDNA CO I gene were sequenced successfully by sequencing directly from a total of 27 samples including 15 A.glabripennis samples from different areas , 4 A.nobilis samples from different areas and the other 4 sibling species of A.glabripennis.From the result of analysis on the sequences, it can be seen that there are base difference in A.glabripennis samples from different areas and show definitive regulation. This show that there is visible marker difference in A.glabripennis from America, China and Korea. This indicates that A.glabripennis from America have formed unique character themselves and can differentiate from A.glabripennis from China and Korea. So we infer the A.glabripennis distributed in U.S.A. were not from China.On the basis of the base difference and the Bootstrap tree, The A.glabripennis specimens including A.nobilis of this study can be divided into 4 populations: the population of America, the population of Korea, the population of the North of China and that of the South of China. The population of the North of China consist of A.glabripennis from the provinces of Gansu, Neimenggu, Hebei and Shananxi. The population of the South of China consist of A.glabripennis from Anhui, Jiangsu, Zhejiang, Hubei, Henan, Shandong, Ningxia, Shanxi and Liaoning.The base difference between A.glabripennis and A.nobilis samples is very small. And they all grouped in a clade. Thus show these two sibling species should belong to the same species.The morphological characters of are very similar. As a result, the biggest base difference between A.glabripennis samples and A.freyi from Chongqing is 6.8% and they were grouped into tow clades in the Bootstrap tree. Therefore, A.freyi can be seen a independent species in the light of the result.The result of sequencing indicates the region of mtDNA CO I gene show distinct difference in different species but conservation in A. glabripennis samples relatively. This regionhas special stability and specificity. It can be used to identify A.glabripennis by sequencing directly.A pair of specific PCR primers was designed according to the sequences of mtDNA CO I gene had sequenced. These primers can be used to identify A.glabripennis. The PCR system and condition of specific PCR of identifying A.glabripennis were optimized and defined the optimum PCR system and condition. A total of 39 specimens were tested including 21 A.glabripennis specimens of different geographical sources. The result show that the primers designed have high specificity and sensitivity. The identification kit for A.glabripennis and its working protocol were developed primarily based on the PCR and have obtained the register patent.