cDNA representational difference analysis of deltamethrin-resistant and susceptible strains of diamondback moth
2013
Zhang, H. (Nanjing Normal Univ., Nanjing (China). Coll. of Life Science) | Feng-Liang, L. (Guizhou Academy of Agriculture Sciences, Guiyang (China). Inst. of Plant Protection) | Cheng, C. (Nanjing Normal Univ., Nanjing (China). Coll. of Life Science) | Bai-Duo, L. | Luo-Gen, C.
The diamondback moth (Plutella xylostella L.), a universal insect pest of cruciferous crops, has shown an extraordinary capacity to resist synthetic pesticides. Identification of genes associated with insecticide resistance has been a valuable tool to understand the mechanisms of resistance to insecticides. cDNA representational difference analysis (cDNA RDA) was performed to screen the upregulated genes in deltamethrin-resistant samples compared with the controls (susceptible strain). The fourth instar larve of deltamethrin-resistant and -susceptible strains of diamondback moth were selected for this study. The cDNA of resistant strain was used as tester amplicon, the driver amplicon was from the susceptible strain. After three rounds of substractive hybridization, we obtained six differentially expressed genes. Comparison between our results and data in the GenBank showed that one differential sequence had high homology with ribosomal protein gene L7 (RpL7). To validate the result, RpL7 was cloned and RpL7 expression levels between the resistant samples and the controls were detected using real-time quantitative PCR (gPCR). The results showed that mRNA level of RpL7 was significantly higher in deltamethrin-resistant samples than in the controls. There are no direct reports indicating the upregulated gene RpL7 is associated with insecticide resistance, further studies are still needed.
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