[Objective] The paper was to improve the efficiency and accuracy of early forecast of Lepidopteran oak-infesting pests.[Method] DNA barcoding technique was established for quick species identification using mitochondr...[Objective] The paper was to improve the efficiency and accuracy of early forecast of Lepidopteran oak-infesting pests.[Method] DNA barcoding technique was established for quick species identification using mitochondrial cytochrome C oxidase subunit Ⅰ（COⅠ） as the standard gene.This barcoding technique was used to amplify and sequence genomic DNA samples from eggs and pupae of 11 species of Lepidopteran pests collected from oak.[Result] The DNA barcoding standard genes of 594-708 bp were determined from eggs and pupae of Lepidopteran insects.There were differences of 0-2 bases in DNA barcode sequences between conspecific eggs and pupae,with the sequence identity of 99.7%-100%.The average content of A,T,G and C of DNA barcode sequences from Lepidopteran insects were 30.7%,38.5%,14.9% and 15.9%,respectively.The obtained DNA barcode sequences had 91.4%-100% identity and 0-8.6% difference degree with GenBank-deposited DNA barcode sequences from organisms of the genetically-closest relationship.Among them,DNA barcode sequences from egg and pupa samples of 10 Lepidopteran insects（No.1-20） had 99%-100% identity and 0-1.0% difference degree with homologous sequences in GenBank database,while the remaining samples（No.21-22） had high difference degree（8.6%） with homologous sequences.[Conclusion] The established DNA barcoding technique is an effeetive tool for species identification of Lepidopteran pests using genomic DNA from eggs and pupae of Lepidopteran insects.展开更多
基金Supported by Natural Science Foundation of Liaoning Province(2014027002)National Special Fund for Development of Cocoon Silk(201601)National Modern Agriculture Industry Technology System Construction Project(CARS-18)
文摘[Objective] The paper was to improve the efficiency and accuracy of early forecast of Lepidopteran oak-infesting pests.[Method] DNA barcoding technique was established for quick species identification using mitochondrial cytochrome C oxidase subunit Ⅰ（COⅠ） as the standard gene.This barcoding technique was used to amplify and sequence genomic DNA samples from eggs and pupae of 11 species of Lepidopteran pests collected from oak.[Result] The DNA barcoding standard genes of 594-708 bp were determined from eggs and pupae of Lepidopteran insects.There were differences of 0-2 bases in DNA barcode sequences between conspecific eggs and pupae,with the sequence identity of 99.7%-100%.The average content of A,T,G and C of DNA barcode sequences from Lepidopteran insects were 30.7%,38.5%,14.9% and 15.9%,respectively.The obtained DNA barcode sequences had 91.4%-100% identity and 0-8.6% difference degree with GenBank-deposited DNA barcode sequences from organisms of the genetically-closest relationship.Among them,DNA barcode sequences from egg and pupa samples of 10 Lepidopteran insects（No.1-20） had 99%-100% identity and 0-1.0% difference degree with homologous sequences in GenBank database,while the remaining samples（No.21-22） had high difference degree（8.6%） with homologous sequences.[Conclusion] The established DNA barcoding technique is an effeetive tool for species identification of Lepidopteran pests using genomic DNA from eggs and pupae of Lepidopteran insects.
