CO2 enhances effects of hypoxia on mortality, development, and gene expression in cowpea bruchid, Callosobruchus maculatus

Modified atmosphere based on lack of O2 offers a safe, residue-free alternative to chemical fumigants for pest control in stored grains. In this study, we intended to determine whether elevated CO2 (at a biologically achievable level) has an enhanced suppressive effect over low O2 atmosphere alone on the cowpea bruchid (Callosobruchus maculatus), a storage pest of cowpea and other legumes. Experiments were performed under two modified atmospheric conditions, (1) 2% O2 + 18% CO2 + 80% N2 and (2) 2% O2 + 98% N2. Both hypoxic environments significantly affected the development and survival of all insect developmental stages. Eggs were most vulnerable to hypoxia, particularly at the early stage (4–6 h old), surviving only up to a maximum of 2 days in both treatments. These were followed by adults, pupae and larvae, in order of decreasing susceptibility. The 3rd and 4th instar larvae were most resilient to hypoxia and could survive up to 20 days of low O2. The presence of 18% CO2 significantly increased the mortality of adults, the later stage of eggs, as well as 1st and 4th instar larvae caused by hypoxia. However, the surviving insects exhibited faster development, evidenced by their earlier emergence from cowpea seeds compared to those without CO2. One interesting observation was the frequent, premature opening of the emergence windows in the 4th instar larvae when CO2 was involved. This phenomenon was not observed at all in insects stressed by low O2 alone. Differential expression profiling of metabolic genes and proteolytic activity of midgut digestive enzymes suggested that the rate of metabolic activity could contribute in part to the difference in insect development and survival under hypoxia in the presence and absence of CO2.