利用RNAi技术沉默麦长管蚜与桃蚜细胞色素P450

doi:10.3864/j.issn.0578-1752.2012.17.002

Abstract

Abstract: 【Objective】Silencing of cytochrome P450 in S. avenae and M. persicae through RNA interference by feeding on artificial diet containing double-stranded RNA (dsRNA).【Method】 Fragments of cytochrome P450 in S. avenae and M. persicae were obtained by PCR. The dsRNA of the conserved sequence of cytochrome P450 from S. avenae and M. persicae were synthesized by dsRNA synthesis kit. dsRNA was added to the final concentration of 0, 3, 5, and 7.5 ng•μL-1 in aphid artificial diet, respectively. Three replicates were set up for each experimental group. The number of surviving aphids was counted after 2, 4, 6, and 8 days of feeding. The expression level of target genes were analyzed by real-time fluorescence quantitative PCR.【Result】Cytochrome P450 fragments were cloned from the cDNA of S. avenae and M. persicae, the homology of the two genes was 90.1%. The mortality of aphids increased along with the increased dsRNA concentration as time went on. Total RNA were extracted from aphids fed by dsRNA at concentration of 7.5 ng•μL-1 and collected from different time periods. The results of real-time fluorescence quantitative PCR showed that the target genes were silenced significantly. 【Conclusion】The expression level of cytochrome P450 in S. avenae and M. persicae which fed by the dsRNA of the conserved sequences of cytochrome P450, were suppressed, and thus led to the death of aphids.

Key words: Sitobion avenae, Myzus persicae, cytochrome P450, RNAi, dsRNA

WANG Hui, ZHANG Min, ZHANG Xiao-Hong, XIA Lan-Qin. Silencing of Cytochrome P450 in Sitobion avenae and Myzus persicae Through RNA Interference[J].Scientia Agricultura Sinica, 2012, 45(17): 3463-3472.

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Silencing of Cytochrome P450 in Sitobion avenae and Myzus persicae Through RNA Interference

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