Novel regulation pathway of eclosion hormones in Tribolium castaneum by distinct transcription factors through the initiation of 20-Hydroxyecdysone.

Eclosion hormone (EH) is not only a key trigger of insect ecdysis, but is also involved in the regulation of important physiological processes such as development, diapause, metamorphosis and reproduction. EH is an ideal target for RNAi treatment and prevention of the Tribolium castaneum. However, two EH genes in Tribolium castaneum demonstrate distinct replication and functional conversion relationships, and the mechanisms of transcriptional regulation of eclosion hormone remain largely unexplored and poorly understood. In this study, the activity of highly active promoter fragments and potential transcription factors of TcEH and TcEHL were identified using the Dual-Luciferase reporter system and TANSFAC. TcSlbo and TcCAD were revealed to be important transcription factors for TcEH and TcEHL, respectively. Knockdown of TcSlbo failed to slough off the old epidermis of Tribolium castaneum and prevented them from developing into adults. Furthermore, we demonstrated for the first time that 20-hydroxyecdysone (20E) affects the expression of TcEH and TcEHL by regulating the transcriptional activities of TcSlbo and TcCAD. This study provides new insights into the transcription regulation of TcEH and TcEHL, their roles in insect growth and development, and the involvement of 20E in eclosion regulation, offering potential molecular targets for future pest management strategies.

CRISPR/Cas9-Mediated Knockout of the PxJHBP Gene Resulted in Increased Susceptibility to Bt Cry1Ac Protoxin and Reduced Lifespan and Spawning Rates in Plutella xylostella.

Juvenile hormone binding protein (JHBP) is a key regulator of JH signaling, and crosstalk between JH and 20-hydroxyecdysone (20E) can activate and fine-tune the mitogen-activated protein kinase cascade, leading to resistance to insecticidal proteins from Bacillis thuringiensis (Bt). However, the involvement of JHBP in the Bt Cry1Ac resistance of Plutella xylostella remains unclear. Here, we cloned a full-length cDNA encoding JHBP, and quantitative real-time PCR (qPCR) analysis showed that the expression of the PxJHBP gene in the midgut of the Cry1Ac-susceptible strain was significantly higher than that of the Cry1Ac-resistant strain. Furthermore, CRISPR/Cas9-mediated knockout of the PxJHBP gene significantly increased Cry1Ac susceptibility, resulting in a significantly shorter lifespan and reduced fertility. These results demonstrate that PxJHBP plays a critical role in the resistance to Cry1Ac protoxin and in the regulation of physiological metabolic processes associated with reproduction in adult females, providing valuable insights to improve management strategies of P. xylostella.

Polygenic adaptation of a cosmopolitan pest to a novel thermal environment.

The fluctuation in temperature poses a significant challenge for poikilothermic organisms, notably insects, particularly in the context of changing climatic conditions. In insects, temperature adaptation has been driven by polygenes. In addition to genes that directly affect traits (core genes), other genes (peripheral genes) may also play a role in insect temperature adaptation. This study focuses on two peripheral genes, the GRIP and coiled-coil domain containing 2 (GCC2) and karyopherin subunit beta 1 (KPNB1). These genes are differentially expressed at different temperatures in the cosmopolitan pest, Plutella xylostella. GCC2 and KPNB1 in P. xylostella were cloned, and their relative expression patterns were identified. Reduced capacity for thermal adaptation (development, reproduction and response to temperature extremes) in the GCC2-deficient and KPNB1-deficient P. xylostella strains, which were constructed by CRISPR/Cas9 technique. Deletion of the PxGCC2 or PxKPNB1 genes in P. xylostella also had a differential effect on gene expression for many traits including stress resistance, resistance to pesticides, involved in immunity, trehalose metabolism, fatty acid metabolism and so forth. The ability of the moth to adapt to temperature via different pathways is likely to be key to its ability to remain an important pest species under predicted climate change conditions.

Thermal acclimation uncovers a simple genetic basis of adaptation to high temperature in a cosmopolitan pest.

Understanding a population's fitness heterogeneity and genetic basis of thermal adaptation is essential for predicting the responses to global warming. We examined the thermotolerance and genetic adaptation of Plutella xylostella to exposure to hot temperatures. The population fitness parameters of the hot-acclimated DBM strains varied in the thermal environments. Using genome scanning and transcription profiling, we find a number of genes potentially involved in thermal adaptation of DBM. Editing two ABCG transporter genes, PxWhite and PxABCG, confirmed their role in altering cuticle permeability and influencing thermal responses. Our results demonstrate that SNP mutations in genes and changes in gene expression can allow DBM to rapidly adapt to thermal environment. ABCG transporter genes play an important role in thermal adaptation of DBM. This work improves our understanding of genetic adaptation mechanisms of insects to thermal stress and our capacity to predict the effects of rising global temperatures on ectotherms.

A very long-chain fatty acid enzyme gene, PxHacd2 affects the temperature adaptability of a cosmopolitan insect by altering epidermal permeability.

Temperature fluctuations pose challenges to poikilotherms, such as insects, especially under climate change conditions. Very long-chain fatty acids (VLCFAs) form important structural components of membranes and epidermal surfaces, so play important roles in adaptation to temperature stress in plants. It has been unclear whether VLCFAs are involved in epidermis formation and thermal resistance in insects. In this study, we focused on the 3-hydroxy acyl-CoA dehydratase 2 (Hacd2), an important enzyme in the synthesis pathway of VLCFAs, in a cosmopolitan pest, the diamondback moth, Plutella xylostella. Hacd2 was cloned from P. xylostella and the relative expression pattern was identified. Epidermal permeability increased with the decreased VLCFAs in the Hacd2-deficient P. xylostella strain, which was constructed by using the CRISPR/Cas9 system. Survival and fecundity of the Hacd2-deficient strain was significantly lower than that of the wildtype strain when subject to desiccating environmental stress. Hacd2 mediates thermal adaptability in P. xylostella by changing epidermal permeability so is likely to be key to its remaining a major pest species under predicted climate change conditions.

PxTret1-like Affects the Temperature Adaptability of a Cosmopolitan Pest by Altering Trehalose Tissue Distribution.

Global warming poses new challenges for insects to adapt to higher temperatures. Trehalose is the main blood sugar in insects and plays an important role in energy metabolism and stress resistance. The transmembrane transport of trehalose mainly depends on the trehalose transporter (TRET1). Plutella xylostella (L.) is a worldwide agricultural pest; however, the effects of the trehalose transport mechanism and trehalose distribution in tissues on the development, reproduction and temperature adaptation of P. xylostella have yet to be reported. In this study, PxTret1-like was cloned and analyzed regarding its expression pattern. It was found that the expression of PxTret1-like was affected by ambient temperature. The knockout mutation of PxTret1-like was generated using a CRISPR/Cas9 system by targeted knockout. The trehalose content and trehalase activity of mutant P. xylostella increased at different developmental stages. The trehalose content increased in the fat body of the fourth-instar P. xylostella, and decreased in the hemolymph, and there was no significant change in glucose in the fat body and hemolymph. Mutant strains of P. xylostella showed a significantly reduced survival rate, fecundity and ability to withstand extreme temperatures. The results showed that PxTret1-like could affect the development, reproduction and temperature adaptability of P. xylostella by regulating the trehalose content in the fat body and hemolymph.

Type and distribution of sensilla in the antennae of Euplatypus parallelus (F.) (Coleoptera: Curculionidea, Platypodinae).

Euplatypus parallelus (F.) (Coleoptera: Curculionidea) is the most destructive cosmopolitan insect pest of the Platypodinae. Pheromone-based luring agents are used currently in controlling bark beetle. Antennae are the primary insect organs sensing volatiles of host trees and pheromones of pioneer males. We studied the external morphology of antennae and the type, distribution, and the number of the beetle sensilla. Our results show E. parallelus have a geniculate antenna composed of 6 segments, namely the scape, 4-segmented funicle and club. Ninety-seven percent of the antennal sensors were distributed in the club, and 3% were distributed in the scape and funicle. 6 types of sensilla on the antennae were found, including sensilla trichodea (subtypes: STI, STII and STIII), sensilla basiconica (subtypes: SBI, SBII, SBIII and SBIV), sensilla chaetica (subtypes: SChI, SChII and SChIII), as well as sensilla coeloconica, sensilla campaniform and sensilla furcatea. There was no significant difference in the type, distribution and number of sensilla in males and females. No significant difference in the shape and distribution of antennae was found between sexes, but the length of antennae and the number of SChI, SChII, STI, SBI, SBIII and SBIV were significantly larger in females than males. We revealed the external cuticular structure of the antennae in E. parallelus, which can be used to guide future electrophysiological investigations to understand the ability of this beetle to detect semiochemicals.

Fine structure of mouthparts and forelegs of Euplatypus parallelus (Coleoptera: Curculionidae) with emphasis on the behavior of gallery excavation.

Euplatypus parallelus (F.) (Coleoptera: Curculionidae) is one of the most invasive species of all the Platypodinae. It penetrates the xylem and oviposits in its host trees thereby weakening the trunk causing them to break under extreme conditions. Since the beetle has evolved effective drilling mouthparts enough to make wood tunnels, we used a field emission scanning electron microscopy to describe the sexual difference in mouthparts and forelegs morphology of the beetle. E. parallelus has chewing type mouthparts composed of a labrum, a pair of mandibles, a pair of maxillae, and a labium. In females, the size of maxillary palpi, submentum, prementum, and labial palpi are significantly larger than males. E. parallelus forelegs were walking type composed of procoxa, protrochanter, profemur, protibia, protarsus, and propretarsus. We observed no significant differences between the forelegs of males and females, but the procoxa of the males was slightly larger than that of females. The structural differences in mouthparts and forelegs between females and males indicated that females invest more time in gallery excavation than males. Possible functional relationships of these structures are discussed. These studies revealed the mechano-dynamic characteristics of E. parallelus and provided a theoretical basis for exploring the behavior of this beetle.