Revision as of 19:35, 23 September 2018

Experimental summary

Short interfering RNAs (siRNAs) and their corresponding shRNAs were designed based on the mRNA sequences of their target genes. siRNAs were synthesized by a bio-company, shRNAs were produced by in vitro transcription. The efficiency of both siRNAs and shRNAs in mediating RNAi in Phyllotreta striolata were examined. Experimental results show that both siRNAs and shRNA could successfully silence their target genes, which was demonstrated by the survival rate decrease after siRNA or shRNA treatment. Results also show that GC percentage of the total siRNA/shRNA, the 5’ end of the siRNA/shRNA, the 3’ end of the siRNA/shRNA affect the RNAi efficiency.

In addition, we tested the attraction effect of sucrose and lemon yellow on P. S. Results show that sucrose plus lemon yellow has the best attraction effect.

Objectives

The objective of our project is to trigger the RNAi mechanism in Phyllotreta striolata, which could lead to the death of the beetle, by topical application of exogenous shRNA/siRNAs.

1. Target mRNA selection

Based on the Phyllotreta striolata transcriptome sequence data provided by Professor Weichang Yu, we selected our target mRNAs, which are the mRNAs of Arginine Kinase, Glutathione S-Transferase and Aldose Reductase. These three genes encode important enzymes that are involved in metabolic pathways. The mRNA sequences for arginine kinase (Fig. 1-1), Glutathione S-Transferase(Fig. 1-2), and Aldose Reductase(Fig. 1-3) were obtained.