Topical Application of Synthetic Hormones Terminated Reproductive Diapause of a Univoltine Weed Biological Control Agent

Classical biological control is an important method for controlling invasive alien weeds. Univoltine insects can be highly effective biological control agents of annual weeds because they are well synchronized with their host plant. However, having only one generation per year makes it difficult and slow to multiply them in the laboratory for initial field releases. If it were possible to terminate reproductive diapause early, then we could rear multiple generations per year, which would greatly increase annual production. We used a recently approved biocontrol agent, Ceratapion basicorne (a univoltine weevil), for yellow starthistle (Centaurea solstitialis) as a model system to study the use of two insect hormones, 20-hydroxyecdysone (20E) and methoprene, to terminate reproductive diapause. Methoprene (1 μg applied topically) terminated reproductive diapause of female weevils, whereas doses of 0.0, 0.01 and 0.1 μg did not. The combination of methoprene and 20E had a stronger effect and induced an increase in eggs (1.51 ± 0.16 eggs/day, mean ± SE) compared with a methoprene only group (1.00 ± 0.13 eggs/day), and a control group (0.21 ± 0.04 eggs/day). Thus, topical application of these hormones should enable us to rear the weevil out of its normal season and produce more than one generation per year, which will increase productivity of mass-rearing it for field release. Once released in the field, the insect would continue as a univoltine agent that is well-synchronized with its host plant.

Regulation of Juvenile Hormone on Summer Diapause of Geleruca daurica and Its Pathway Analysis

Juvenile hormone (JH) signaling plays an important role in regulation of reproductive diapause in insects. However, we have little understanding of the effect of JH on gene expression at the transcriptome level in diapause. Galeruca daurica is a new pest in the Inner Mongolia grasslands with obligatory summer diapause in the adult stage. Topical application of a JH analog methoprene at the pre-diapause stage delayed the adults entering diapause and inhibited lipid accumulation whereas it did not during diapause. Using Illumina sequencing technology and bioinformatics tools, 54 and 138 differentially expressed genes (DEGs) were detected at 1 and 2 d after treatment, respectively. The KEGG analysis showed that the DEGs were mainly enriched in the metabolism pathways. qRT-PCR analysis indicated that methoprene promoted the expression of genes encoding vitellogenin, fork head transcription factor and Krüppel homolog 1, whereas suppressed the expression of genes encoding juvenile hormone-binding protein, juvenile hormone esterase, juvenile hormone acid methyltransferase, juvenile hormone epoxide hydrolase and fatty acid synthase 2. These results indicate that JH signaling plays an important role in regulating reproductive diapause of G. daurica.

On the Reproductive Diapause in the Predatory Ladybird Cheilomenes sexmaculata (Fabricius, 1781) (Coleoptera, Coccinellidae)

The ability to enter reproductive diapause was experimentally studied in females of the laboratory population of the predatory ladybird Cheilomenes sexmaculata originated from individuals collected in Nepal in a region of subtropical monsoon climate. The experiment included 12 regimes, i.e. combinations of 2 temperatures (20 and 24°C), 3 photoperiods (day lengths of 10, 12, and 14 h), and 2 diets (the green peach aphid Myzus persicae and eggs of the grain moth Sitotroga cerealella). Females with undeveloped ovaries and well developed fat body were considered as diapausing. The proportion of diapausing females was significantly dependent only on the temperature being 0.8% at 24°С and 7.4% at 20°С. Although found in only a small fraction of the studied population, the ability to enter reproductive diapause (as suggested by literature data) was probably an important prerequisite for Ch. sexmaculata spread northwards to the temperate zone.

Steroid hormone ecdysone deficiency stimulates preparation for photoperiodic reproductive diapause

Diapause, a programmed developmental arrest primarily induced by seasonal environmental changes, is very common in the animal kingdom, and found in vertebrates and invertebrates alike. Diapause provides an adaptive advantage to animals, as it increases the odds of surviving adverse conditions. In insects, individuals perceive photoperiodic cues and modify endocrine signaling to direct reproductive diapause traits, such as ovary arrest and increased fat accumulation. However, it remains unclear as to which endocrine factors are involved in this process and how they regulate the onset of reproductive diapause. Here, we found that the long day-mediated drop in the concentration of the steroid hormone ecdysone is essential for the preparation of photoperiodic reproductive diapause in Colaphellus bowringi, an economically important cabbage beetle. The diapause-inducing long-day condition reduced the expression of ecdysone biosynthetic genes, explaining the drop in the titer of 20-hydroxyecdysone (20E, the active form of ecdysone) in female adults. Application of exogenous 20E induced vitellogenesis and ovarian development but reduced fat accumulation in the diapause-destined females. Knocking down the ecdysone receptor (EcR) in females destined for reproduction blocked reproductive development and induced diapause traits. RNA-seq and hormone measurements indicated that 20E stimulates the production of juvenile hormone (JH), a key endocrine factor in reproductive diapause. To verify this, we depleted three ecdysone biosynthetic enzymes via RNAi, which confirmed that 20E is critical for JH biosynthesis and reproductive diapause. Importantly, impairing Met function, a component of the JH intracellular receptor, partially blocked the 20E-regulated reproductive diapause preparation, indicating that 20E regulates reproductive diapause in both JH-dependent and -independent manners. Finally, we found that 20E deficiency decreased ecdysis-triggering hormone signaling and reduced JH production, thereby inducing diapause. Together, these results suggest that 20E signaling is a pivotal regulator that coordinates reproductive plasticity in response to environmental inputs.