The Function of LmPrx6 in Diapause Regulation in Locusta migratoria Through the Insulin Signaling Pathway

SIMPLE SUMMARY: LmPrx6 of the insulin signaling pathway is significantly associated with diapause induction in Locusta migratoria L. as per our pervious transcriptome data. In the current study, we first cloned and sequenced the gene and demonstrated its similarity to other Prxs using phylogenetic analyses. Later on, we knocked down Prx6 using RNAi and showed that phosphorylation of proteins associated with the insulin signaling pathway and responses to oxidative stress were altered. Knockdown of Prx6 also resulted in a reduced ability to enter diapause, and hence, we are of the opinion that this gene could serve as an effective target for RNAi-based control of L. migratoria L. The study has provided some helpful insights into the diversified roles of Prx6 in locusts and will be of interest to other insect pests for examining the relatively unexplored group of proteins as well. ABSTRACT: Peroxiredoxins (Prxs), which scavenge reactive oxygen species (ROS), are cysteine-dependent peroxide reductases that group into six structurally discernable classes: AhpC-Prx1, BCP-PrxQ, Prx5, Prx6, Tpx, and AhpE. A previous study showed that forkhead box protein O (FOXO) in the insulin signaling pathway (ISP) plays a vital role in regulating locust diapause by phosphorylation, which can be promoted by the high level of ROS. Furthermore, the analysis of transcriptome between diapause and non-diapause phenotypes showed that one of the Prxs, LmPrx6, which belongs to the Prx6 class, was involved. We presumed that LmPrx6 might play a critical role in diapause induction of Locusta migratoria and LmPrx6 may therefore provide a useful target of control methods based on RNA interference (RNAi). To verify our hypothesis, LmPrx6 was initially cloned from L. migratoria to make dsLmPrx6 and four important targets were tested, including protein-tyrosine phosphorylase 1B (LmPTP1B), insulin receptor (LmIR), RAC serine/threonine-protein kinase (LmAKT), and LmFOXO in ISP. When LmPrx6 was knocked down, the diapause rate was significantly reduced. The phosphorylation level of LmPTP1B significantly decreased while the phosphorylation levels of LmIR, LmAKT, and LmFOXO were significantly increased. Moreover, we identified the effect on two categories of genes downstream of LmFOXO, including stress tolerance and storage of energy reserves. Results showed that the mRNA levels of catalase and Mn superoxide dismutase (Mn-SOD), which enhanced stress tolerance, were significantly downregulated after silencing of LmPrx6. The mRNA levels of glycogen synthase and phosphoenolpyruvate carboxy kinase (PEPCK) that influence energy storage were also downregulated after knocking down of LmPrx6. The silencing of LmPrx6 indicates that this regulatory protein may probably be an ideal target for RNAi-based diapause control of L. migratoria.