Study On DNA Barcoding Identification Kit Technologyof Cerambycidae Species From Ports

The insects of Cerambycidae are an important group in the tree trunk, which can damage living trees, and forest product damage, can also be spread through trade in goods over long distances. Some species of tree pathogens or the media, causing serious damage and economic loss. With the rapid development of China’s foreign trade, foreign forest pest Cerambycidae insects especially repeatedly intercepted at ports, that the threat of foreign species of forest ecological environment in our country is growing.Accurate identification of the inspection and quarantine pest species is an important part of the work. The identification of quarantine insect with morphological method, but the traditional morphological identification, there are some limitations, such as the early form of pest insects and incomplete body illegible, the identification results are greatly influenced by subjectivity, appraisal cycle is very long, to a certain extent affected the port quarantine and epidemic detection rate of accuracy. DNA Barcoding is one of the research hotspots of biological identification and classification in recent years, the development of this technology makes up the deficiency of the traditional morphological classification, and provides a new technology for molecular identification of harmful organisms. The research has been widely used in the discovery and description of new species or cryptic species, species identification, phylogeny.This study is based on the mitochondrial COI(mitochondrial cytochrome oxidase subunit I), to the Monochamus, Clytini, Tetropium, Batocera, the feature identification of population control, preliminary obtained genetic characteristics of different kinds of family, genus, which is able to distinguish. The main results are as follows:1. Animal and plant DNA extraction method was developed by Canada CCDB barcode reagent improved extraction of Cerambycidae insect mitochondrial COI gene. The integrity of the extraction method can not only save the precious specimens, and the cost is low, but compared to the now available in the DNA extraction kit to extract the time-consuming disadvantage.2. Based on the development of DNA bar code technology, proposed the insect DNA barcode Kit(DNA barcoding identification kit technology) concept. According to the mitochondrial DNA nucleotide site between closely related species, the sample after PCR amplification and sequencing to obtain a simple marker bases, mark base is simple and clear, as the basis for species identification.3. This study incloud DNA barcode kit includes 24 kinds: Clytini, Tetropium Kirby 9 species, 12 kinds of Monochamus longicorn, Batocera sp. is the 4 important quarantine pest. Taking into account the amplification of specific part of family, genus, we designed a pair of primers were used to amplify the broken nested eye beetle genus and Clytini Sawyer COI gene family.4. Involving Cerambycidae 4 genera in the premise of constructing the phylogenetic tree and sequence analysis of down to 4 in this study, eventually won the tag base, which belong to the species and similar species can be differentiated, provides a new means for the future of quarantine port work.5. Established a quarantine pest database system. In this study, the sequence analysis software from the pest quarantine system, the system includes the port inspection and Quarantine of longicorn, often intercepted beetles, flies and other important quarantine pest damage, biological characteristics, morphological identification, gene sequence and an important quarantine genus, family bar kit.