Studies On The Genetic Transformation Mediated By Piggybac In The Asian Corn Borer, Ostrinia Furnacalis

The Asian corn borer (ACB) is a major pest of corn, millet, sorghum, cotton, and so on. These pests distribute widely in China, from Heilongjiang province to Guangdong province. Nowadays, because of the overuse of pesticides, on the one hand, the environment is destroyed, and the food safety is challenged; for another, resistance of this pest is normal so as to reduction of control efficiency. Therefore, researchers primary focus on a novel, environmentally friendly, and species-specific strategy for controlling ACB.A piggyBac vector was successfully constructed to express enhanced green fluorescence protein (EGFP) under the control of Bombyx mori actin3promoter (A3). Transient EGFP expression was detected at48h after preblastodermic microinjection of the piggyBac vector. The excision assay showed the vector was precisely excised, and transposase mRNA was more effective in activating transposase than pBac[A3Helper] plasmid in ACB embryos. In G1animals, PCR-based investigations revealed that exogenous vector had been introduced into ACB genome and expressed successfully at transcriptional level. In addition, western blot analysis showed that EGFP had expressed efficiently, indicating the heritability of the transgene.To further understand the biological and evolutionary characteristics of piggyBac-like elements (PLEs), PLE sequences, named as OfPLE1and OfPLE2, were cloned from ACB based on degenerate PCR, inverse PCR, and RACE. The sequence structure of OfPLE gene coding region and the phylogenetic relationship with other species were analyzed by application of the bioinformatics software. The results showed that OfPLE1was1866bp long encoding607amino acid residues and OfPLE2was1724bp long encoding434amino acid residues. Furthermore, they shared70%homology with each other, and had32%identity and26%similarity with Trichoplusia ni piggyBac transposon.These results indicated that transient EGFP expression was screened after egg microinjection, and exogenous DNA had stably integrated into genome of transgenic next generations. In addition, we identified two isforms of PLEs. These provide solid evidence for carrying out genetic control in ACB.