Molecular cloning, expression profiling, and functional analysis of a broad-complex isoform 2/3 (Br-Z2/Z3)...

The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a widespread and destructive pest of cruciferous crops. New strategies for controlling it are needed because it is rapidly developing resistance to conventional pesticides. In insects, transcription factors (TFs) including broad-complex (Br-C) are thought to be useful for insecticide development because they are able to regulate the transcription of functional genes involved in responses to external stimuli including insecticides. In the present study, we cloned and sequenced the open reading frames (ORFs) of three BTB-ZF encoding genes from the diamondback moth deposited in the National Center for Biotechnology Information (NCBI) database under accessions MG753773, MG288674, and MG753772. The lengths of these ORFs were 1,680, 1,428, and 1,647 bp, respectively. The phylogenetic analysis based on the predicted amino acid sequences of ZF domains showed that MG753773 and MG288674 belonged to Z2/Z3 and Z7 of Br-C while MG753772 belonged to Ttk types. In the agreement, the highest expression level of MG753773 occurred during the prepupal stage, MG288674 and MG753772 were expressed during all stages and peaked in the adult and egg stages, respectively. RNA interference silencing of MG753773 in the late third instar larvae significantly decreased survival and pupation of the insects. With precocene II, transcription of MG753773 increased (4×) in the fourth instar larva 24 hr later; 48 hr later the rate of prepupation and pupation was significantly higher. These findings will contribute to the development of new regulators of the growth and development for diamondback moth control.