The fall armyworm (FAW), Spodoptera frugiperda, is native to the tropical and subtropical areas of the American continent and is one of the world's most destructive insect pests and invaded Africa and spread to most of Asia in two years. Glycerol is generally used as a cryoprotectant for overwintering insects in cold areas. In many studies, the increase in glycerol as a main rapid cold hardening (RCH) factor and enhancing the supercooling point was revealed at low temperatures. There are two genes, including glycerol-3-phosphate dehydrogenase (GPDH) and glycerol kinase (GK), that were identified as being associated with the glycerol synthesis pathway. In this study, one GPDH and two GK sequences (GK1 and GK2) were extracted from FAW transcriptome analysis. RNA interference (RNAi) specific to GPDH or GK1 and GK2 exhibited a significant down-regulation at the mRNA level as well as a reduction in survival rate when the RNAi-treated of FAW larvae post a RCH treatment. Following a cold period, an increase in glycerol accumulation was detected utilizing high-pressure liquid chromatography and colorimetric analysis of glycerol quantity in RCH treated hemolymph of FAW larvae. This research suggests that GPDH and GK isozymes are linked to the production of a high quantity of glycerol as an RCH factor, and glycerol as main cryoprotectant plays an important role in survival throughout the cold period in this quarantine pest studied.

The red imported fire ant (RIFA), Solenopsis invicta Buren is native to South America and is known as a global problematic invasive species. This study focused on the molecular response of RIFA by comparing gene expression profiles after exposing ants to low (10 °C) and high (40 °C) temperature stress and comparing them to untreated controls (30 °C). A total of 99,085 unigenes (the clustered non-redundant transcripts that are filtered from the longest assembled contigs) were obtained, of which 19,154 were annotated with gene descriptions, gene ontology terms, and metabolic pathways. 86 gene ontology (GO) functional sub-groups and 23 EggNOG terms resulted. Differentially expressed genes (DEGs) with log₂FC ≥ 10 were screened and were compared at different temperatures. We found 203, 48, and 66 specific DEGs co-regulated at 10, 20, and 40 °C. Comparing transcriptome profiles for differential gene expression resulted in various DE genes, including cytochrome P450, NADH dehydrogenase subunit 1, cuticle protein and heat shock protein (HSP), which have previously been reported to be involved in cold and high temperature resistance. GO analysis revealed that antioxidant activity is up-regulated under high temperature stress. We verified the RNA-seq data by qPCR on 20 up- and down-regulated DEGs. These findings provide a basis for future understanding of the adaptation mechanisms of RIFA and the molecular mechanisms underlying the response to low and high temperatures.

In this study, we analyzed the potential distribution of red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), in response to climate change in South Korea using CLIMEX, a species distribution model. We further attempted to evaluate the risk of the distribution/invasion and subsequent dispersion by considering climatic suitability, and functional characteristics of cities and covered cultivated areas. The climatic suitability has extended from the southern and coastal regions to inland regions due to climate change. The number of areas with EI (Ecoclimatic Index) values of more than 20 was 9 (12%) in the current climate; the value was assumed to increase to 23% (2040), 24% (2060), 42% (2080), and 62% (2100) from the South Korea coast to inland. We predicted that May to October would be the most active period in seven domestic high-habitation areas. We also analyzed the invasive risk of the red imported fire ant into covered domestic cultivation areas. Considering climatic suitability, we determined that Jeju, Pohang, Busan, Ulsan, Mokpo, and Gosan would be the most affected areas. This study can provide baseline data for the management of invasive species nationally and for regional control through predictions of the probability of settlement and direction of spread.

Insect immunity defends against the virulence of various entomopathogens, including Bacillus thuringiensis (Bt). This study tested a hypothesis that any suppression of immune responses enhances Bt virulence. In a previous study, the entomopathogenic bacterium, Xenorhabdus hominickii (Xh), was shown to produce secondary metabolites to suppress insect immune responses. Indeed, the addition of Xh culture broth (XhE) significantly enhanced the insecticidal activity of Bt against S. exigua. To analyze the virulence enhanced by the addition of Xh metabolites, four bacterial secondary metabolites were individually added to the Bt treatment. Each metabolite significantly enhanced the Bt insecticidal activity, along with significant suppression of the induced immune responses. A bacterial mixture was prepared by adding freeze-dried XhE to Bt spores, and the optimal mixture ratio to kill the insects was determined. The formulated bacterial mixture was applied to S. exigua larvae infesting Welsh onions in a greenhouse and showed enhanced control efficacy compared to Bt alone. The bacterial mixture was also effective in controlling other Spodopteran species such as S. litura and S. frugiperda but not other insect genera or orders. This suggests that Bt+XhE can effectively control Spodoptera-associated pests by suppressing the immune defenses.

Solenopsis japonica, as a fire ant species, shows some predatory behavior towards earthworms and woodlice, and preys on the larvae of other ant species by tunneling into a neighboring colony’s brood chamber. This study focused on the molecular response process and gene expression profiles of S. japonica to low (9 °C)-temperature stress in comparison with normal temperature (25 °C) conditions. A total of 89,657 unigenes (the clustered non-redundant transcripts that are filtered from the longest assembled contigs) were obtained, of which 32,782 were annotated in the NR (nonredundant protein) database with gene ontology (GO) terms, gene descriptions, and metabolic pathways. The results were 81 GO subgroups and 18 EggNOG (evolutionary genealogy of genes: Non-supervised Orthologous Groups) keywords. Differentially expressed genes (DEGs) with log₂fold change (FC) > 1 and log₂FC < −1 with p-value ≤ 0.05 were screened for cold stress temperature. We found 215 unigenes up-regulated and 115 unigenes down-regulated. Comparing transcriptome profiles for differential gene expression resulted in various DE proteins and genes, including fatty acid synthases and lipid metabolism, which have previously been reported to be involved in cold resistance. We verified the RNA-seq data by qPCR on 20 up- and down-regulated DEGs. These findings facilitate the basis for the future understanding of the adaptation mechanisms of S. japonica and the molecular mechanisms underlying the response to low temperatures.