In April, according to the instructions for the use of plasmid pK18mobsacB, we successfully integrated the third upstream and downstream fragment of ribozyme of P. chlororaphis B3-3G into plasmid pK18mobsacB, and successfully obtained knockout plasmid PK18-2.

In May, we first used plasmid PK18-2 to knock out RNAseIII of P. chlororaphis B3-3G, and performed PCR verification and sequencing verification. Subsequently, we introduced pFLP2 plasmid and recombined kana resistance gene.

In June, we constructed the KC1 plasmid expressing actin-dsRNA. In order to exclude the possible influence of non-target gene dsRNA, we also constructed KC2 plasmid expressing gfp-dsRNA.

In July, in order to control the expression of KC1 and KC2 plasmids, we introduced LacI operator at one end of the tac promoter and transformed it into inducable dsRNA expression plasmids KC3 and KC4.

In August, we further successfully integrated the KillerRed protein expression frame on the basis of vectors KC3 and KC4, respectively, and successfully constructed the KC5 and KC6 plasmids

In September, we performed a biological assay and qRT-PCR to detect the expression level of actin gene in Plagiodera versicolora.