原创力文档rna interference of four genes in adult bactrocera dorsalis by feeding their dsrnas四个基因的rna干扰在成人bactrocera背的喂养他们的极.pdfVIP
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rna interference of four genes in adult bactrocera dorsalis by feeding their dsrnas四个基因的rna干扰在成人bactrocera背的喂养他们的极
RNA Interference of Four Genes in Adult Bactrocera
dorsalis by Feeding Their dsRNAs
Xiaoxue Li, Mingyan Zhang, Hongyu Zhang*
State Key Laboratory of Agricultural Microbiology, Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control and Institute of Urban and
Horticultural Pests, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, People’s Republic of China
Abstract
Background: RNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner.
Recently silencing the target gene through feeding has been successfully carried out in many insect species.
Methodology/Principal Findings: Escherichia coli strain HT115 was genetically engineered to express dsRNA targeting
genes that encode ribosomal protein Rpl19, V type ATPase D subunit, the fatty acid elongase Noa and a small GTPase
Rab11. qRT-PCR showed that mRNA level of four target genes was reduced compared to ds-egfp control by feeding either
engineered bacteria or dsRNAs. The maximum down-regulation of each gene varied from 35% to 100%. Tissue specific
examination indicated that RNAi could be observed not only in midgut but also in other tissues like the ovary, nervous
system and fat body. Silencing of rab11 through ingestion of dsRNA killed 20% of adult flies. Egg production was affected
through feeding ds-noa and ds-rab11 compared to ds-egfp group. Adult flies were continuously fed with dsRNA and
bacteria expressing dsRNA for 14 days and up-regulations of target genes were observed during this process. The
transcripts of noa showed up-regulation compared to ds-egfp control group in four tissues on day 7 after continuous
feeding either dsRNA or engineered bacteria. The maximum over-
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