Identification Of Major Stored-product Pests(Coleoptera: Tenebrionidae) Based On DNA Barcoding

The insect,belonged to the family of Tenebrionidae in Coleoptera,which are one kind of the most economically important pests around the world.It will not only directly feed grain,but also endanger the quality of food.What’s the worst,the insect will spead some diaeases,which will lead a great harm for the production and living of human.Correctly identified the pests is the most important step in integrated pest management.However,the traditional method can’t identification of the insect fragments,immature stages.Therefore,it is urgent to identify the non-adult insects by molecular technology.In this study,the molecular technology was applied to identify the stored-product insect.The main results are as follows:1)The knowlege dataset of the stored-product insects(Coleoptera: Tenebrionidae)was built.14 species and 9 genera insect was included.The knowledge database contains the Chinese name of species,Latin name,biological characteristics,distribution,control measures,etc.2)A DNA barcoding database of the stored-product insects(Coleoptera: Tenebrionidae)was built.It was included 116 species in 6 genera from different location.Taking the stored-product insects(Coleoptera: Tenebrionidae),the COI sequence was obtained by using the PCR technique.The validity of the mitochondrial cytochrome c oxidase subunit I(COI)for insect identification was analyzed using phylogenetic tree,Kimura 2-parameter distances.The result showed that these DNA sequences can be used to build a DNA barcode library of the stored-product insects(Coleoptera: Tenebrionidae).3)Species-specific PCR for the identification of three stored-product insect of Genus Palorus(Coleoptera: Tenebrionidae)was built.Based on the DNA sequence,speciesspecific primers for three Palorus species(Palorus subdepressus,Palorus ratzeburgi,Palorus cerylonoides)were designed,and the sensitivity for the species-specific primers was tested.The result showed that species-specific primers can effectively identified the Palorus species.The lower limit was 0.1 ng/μL for P.cerylonoides,the lower limit was was 0.001 ng/μL for the P.ratzeburgi and P.subdepressus.4)Two molecular techniques namely DNA barcoding and species-specific PCR were applied to identify the stored-product insects(Coleoptera: Tenebrionidae).The results show that the two techniques can effectively identified stored-product insects(Coleoptera: Tenebrionidae).In this study,the molecular identification technology of the stored-product insects(Coleoptera: Tenebrionidae)was established based on the knowlege dataset and DNA barcoding database.It provide foundation research for the stored-product insects(Coleoptera: Tenebrionidae).species-specific primers for Palorus identification were designed.It realize the rapid,accurate identification of three stored-product insect of Genus Palorus(Coleoptera: Tenebrionidae).