Integration of transcriptome and proteome reveals molecular mechanisms underlying stress responses of the cutworm, Spodoptera litura, exposed to different levels of lead (Pb).

Heavy metals are major environmental pollutants that affect organisms across different trophic levels. Herbivorous insects play an important role in the bioaccumulation, and eventually, biomagnification of these metals. Although effects of heavy metal stress on insects have been well-studied, the molecular mechanisms underlying their effects remain poorly understood. Here, we used the RNA-Seq profiling and isobaric tags for relative and absolute quantitation (iTRAQ) approaches to unravel these mechanisms in the polyphagous pest Spodoptera litura exposed to lead (Pb) at two different concentrations (12.5 and 100 mg Pb/kg; PbL and PbH, respectively). Altogether, 1392 and 1630 differentially expressed genes (DEGs) and 58, 114 differentially expressed proteins (DEPs) were identified in larvae exposed to PbL and PbH, respectively. After exposed to PbL, the main up-regulated genes clusters and proteins in S. litura larvae were associated with their metabolic processes, including carbohydrate, protein, and lipid metabolism, but the levels of cytochrome P450 associated with the pathway of xenobiotic biodegradation and metabolism were found to be decreased. In contrast, the main up-regulated genes clusters and proteins in larvae exposed to PbH were enriched in the metabolism of xenobiotic by cytochrome P450, drug metabolism-cytochrome P450, and other drug metabolism enzymes, while the down-regulated genes and proteins were found to be closely related to the lipid (lipase) and protein (serine protease, trypsin) metabolism and growth processes (cuticular protein). These findings indicate that S. litura larvae exposed to PbL could enhance food digestion and absorption to prioritize for growth rather than detoxification, whereas S. litura larvae exposed to PbH reduced food digestion and absorption and channelized the limited energy for detoxification rather than growth. These contrasting results explain the dose-dependent effects of heavy metal stress on insect life-history traits, wherein low levels of heavy metal stress induce stimulation, while high levels of heavy metal stress cause inhibition at the transcriptome and proteome levels.

Accumulation and excretion of zinc and their effects on growth and food utilization of Spodoptera litura (Lepidoptera: Noctuidae).

By exposing larvae of the holometabolous insect Spodoptera litura to the artificial diets supplemented with a range of Zinc (Zn) contents, we investigated Zn ingestion, excretion and accumulation in the insect throughout its life cycle. The effects of Zn stress on the survival, growth and food utilization of S. litura were also determined. Zn concentrations in the body (larvae, pupae, and adults), faeces, exuviates, puparium, eggs increased with the increasing Zn concentrations in the diets, while Zn excretion and accumulation by S. litura in 750 mg/kg Zn treatments was lower than the 600 mg/kg Zn treatment. In the 450 mg/kg Zn treatment, the Zn accumulation in S. litura at different developmental stages differed as follows: larvae > pupa > adult. S. litura ingested Zn via feeding and could excrete most of the Zn via faeces (compared with Zn excretion via exuviates) to reduce its internal Zn accumulation (compared with Zn ingestion). Survival and weight were significantly inhibited, and the prolonged period of development (larvae, pupae) and shortened longevity of adults were found in S. litura exposed to Zn stress greater than 450 mg Zn/kg. In the 150-450 mg/kg Zn treatments, the 6th instar larvae increased their relative consumption rate (RCR) and approximate digestibility (AD) (namely, food eaten) to gain weight, which resulted in greater Zn accumulation in the body. Therefore, below the threshold level (being close to 450 mg/kg Zn), S. litura seemed to have a strong homeostatic adjustment ability (increase the amount of food eaten, thereby increasing AD, RCR and Zn excretion via faeces and exuviates) to sustain their weight, and Zn was beneficial and harmless. Although larvae treated with 750 mg/kg Zn had a similar RCR and AD as the control, a reduced weight gain and prolonged larval period resulted in significantly lower relative growth rate (RGR), which indicated surviving insects may allocate more energy from foods for detoxification than for growth.