Helicoverpa assulta (Lepidoptera: Noctuidae) exhibits a specialized herbivorous diet, primarily targeting select Solanaceae plants. Despite its significant economic impact as a pest, causing substantial harm to crops like hot pepper and tobacco, it has received comparatively limited attention in research compared to its generalist counterparts, H. armigera and H. zea.We introduce a chromosome level genome assembly using a Korean H. assulta (Pyeongchang strain, K18). This assembly was achieved through a combined approach utilizing Nanopore long-read sequencing (approximately 78X coverage) and Illumina NovaSeq short-read sequencing (approximately 54X coverage). The total assembled genome spans 424.36 Mb, designated as ASM2961881v1, comprises 62 scaffolds, with 98.7% of the genome contained within 31 scaffolds, confirming the insect's chromosome count (n = 31). The completeness of the assembly is reflected in BUSCO assessment, with values reaching 99.0%, while the repeat content accounts for 33.01%, and 18,593 CDS were annotated. Additionally, 137 genes were identified within 15 orthogroups that have rapidly expanded in H. assulta, while 149 genes in 95 orthogroups have rapidly contracted. This genome draft serves as a valuable resource to explore various aspects of the specialist's biology, enabling research into host-range evolution, chemical communication, insecticide resistance, and comparative investigations with other Heliothine species.

애긴노린재는 긴노린재과에 속하며 한국을 포함한 동아시아 국가의 다양한 곡물 및 관상용 식물의 주요 해충으로 여겨진다. 본 연구에서는 애긴노린재의 17,367 bp 미토콘드리아 유전체에서 13개의 protein-coding genes, 22개의 transfer RNA genes, 2개의 ribosomal RNA genes 과 non-coding A+T rich region를 확인하였다. G+C content는 23%로 나타났고 다른 긴노린재과와의 염기서열 유사성이 N. cymoides (94.5%), N. fuscovittatus (91.7%)으로 높은 것을 발견하였다. 애긴노린재의 미토콘드리아 유전체 정보는 향후 긴노린재과의 진화 연구와 해충 방제를 위한 정보로 널리 사용될 수 있다.

A novel oxidant fumigation (NOF) is a commercial bleaching and disinfection agent. Recent study indicates its insecticidal activity. However, its exact mode of action to kill insects is not known. This study sets up a hypothesis that reactive oxygen species released from NOF is a main factor to kill insects. Plodia interpunctella is a lepidopteran insect pest infesting various stored grains. Both larvae and adults were susceptive to NOF. To test the hypothesis, we needed to identify antioxidant genes in P. interpunctella. Superoxide dismutase (SOD) and thioredoxin-peroxidase (Trx) were identified from P. interpunctella EST library using ortholog sequences of Bombyx mori. Both SOD and Trx were expressed in larvae of P. interpunctella expecially against oxidative stress induced by bacterial challenge. The bacterial challenge also induced some heat shock protein (HSP) genes. Similarly, different doses of NOF significantly induced both SOD and Trx genes. There results suggest that NOF at sublethal doses releases reactive oxygen species, which may be detoxified by the antioxidant activities of SOD and Trx of P. interpunctella.

Prostaglandins (PGs) mediate insect immune responses. However, their biosynthesis in insects is little understood due to lack of cyclooxygenase (COX) ortholog. This study aimed to identify PG-biosynthetic factor(s) in Spodoptera exigua, which has been a well-known insect in possessing immune responses mediated via PGs. Peroxidases (POXs) are a sister group of COX genes. Ten putative POXs (POX-A ∼POX-J) were expressed in S. exigua. Especially, expressions of POX-F and POX-H were inducible to bacterial challenge and expressed in hemocytes and fat body. Individual RNA interference (RNAi) of each of ten POXs was performed by hemocoelic injection of their specific double-strnaded RNAs (dsRNAs). Only RNAi of POX-F or POX-H specifically suppressed hemocyte-spreading behavior and nodule formation. Addition of PGE2 significantly nescued the immunosuppression in either dsRNA treatment of POX-F or POX-H. Structural analysis indicated that both POX-F and POX-H have conserved domain and residues corresponding to peroxinectin of Drosophila melanogaster, which mimics COX-like activity. These results suggest that POX-E and POX-H are involved in PG biosynthesis in S. exigua.

Immune mediators play crucial roles in amplifying the emergency signals with massive amounts of de novo synthesized mediators and relaying the specific recognition signals to the immune-associated target tissues. Eicosanoids are the representative immune mediators and synthesized from a polyunsaturated fatty acid (PUFA), arachidonic acid. Compared to mammalian systems, insects have relatively low levels of arachidonic acid in the biological membranes. This has raised a fundamental issue that eicosanoids may be not significant in insect system. Our previous chemical analysis suggests that the hemocytes of Spodoptera exigua have less than 5% arachidonic acid. We postulated that S. exigua may store arachidonic acid in other tissues, such as fat body. This analysed fatty acid compositions of two immune-associated tissues using a gas chromatography (GC) eguipped with FID detector or GC-MS. Our analysis of PUFA in the immune tissues suggests that insects maintain a low level of PUFA including arachidonic acid due to its evolutionary origin from the paleozoic era at which the oxygen level was 35%, compared to the present era 21%.

Phospholipase A2 (PLA2) catalyze the committed step for eicosanoid biosynthesis and releases arachidonic acid (AA), which is oxygenated into eicosanoids that mediate immune responses in insects. Thus, any inhibition of PLA2 activity would lead to a significant immuno suppression due to lack of eicosanoids. Among more than 15 families of PLA2s, group Ⅳ cytosolic PLA2 (cPLA2) has been mainly associated with the production of eicosanoids associated with immune responses. However, no cPLA2 has been reported in all invertebrates including insects. AcPLA2 candidate gene (SecPLA2) has been identified from a hemocyte transcriptome of the beet armyworm, Spodoptera exigua. RNA interference of SecPLA2 expression significantly reduced cellular immune responses of hemocytes. When the SecPLA2 was expressed and purified, the recombinant SecPLA2 catalyzed a substrate, phosphoatidyl choline, atsn-2 position. Its catalytic activity was sensitive to pH, temperature, and calciumlevel. Furthermore, there combinant SecPLA2 was specifically sensitive to a cPLA2-specificinhibitor, methyl arachidonyl fluorophosphonate.

Immune defense is indispensible for insect survival. However, uncontrolled and excessive immune responses would be highly detrimental and energy-consuming processes. An insect cytokine, plasmatocyte-spreading peptide (PSP), induces hemocyte-spreading behavior as well as activating phenoloxidase (PO) in the beet armyworm, Spodoptera exigua. A hemocyte transcriptome of S. exigua contains a partial sequence of a putative PSP-binding protein (SePSP-BP). SePSP-BP was expressed in all developmental stages especially in hemocytes and fat body. A quantitative RT-PCR showed that the bacterial infection significantly up-regulated the expression level of SePSP-BP. A double-stranded RNA specific to SePSP-BP (dsRNASePSP-BP) was injected and suppressed SePSP-BP expression even in response to bacterial challenge. The larvae treated with dsRNASePSP-BPsuffered high mortality to infection of nonpathogenic bacteria and prolonged high PO activity after the immune challenge. These results suggest that SePSP-BP may play a role in suppressing immune responses as a negative controller

Insect blood cells, hemocytes, inhibit spreading behavior upon bacterial challenge to perform cellular immune responses. Hemocyte spreading is accomplished by cytoskeleton rearrangement, which is activated by various immune mediators, such as biogenic monoamins, plasmatocyte-spreading peptide(psp), and eicosanoids. However, little is known how these, immune mediators. acitvate hemocyte spreading behavior. A small G protein, Rac1, gene was identified in hemocytes of Spodoptera exiqua. Its expressed in most developmental stages accept egg and expecially expresses in hemocytes and fat body of Larval stage. In response to bacterial challenge, its expression was segnificantly up-regulated. However, RNA inteference (RNAi) of Rac1 expression inhibited hemocyte spreading behavior. under RNAi condition of Rac1, octopamine and psp failed to activate hemocyte spreading behavior. Interestingly, as addition of prostaglandinE2 to the RNAiconditioned Larval rescued the mediation of octopamine and psp. These results indicate that Rac1 is required for mediation of octopamine and psp on hemocytespreading behavior and suggest that Rac1 may activate eicosanoid biosnthesis.

As the immune reactions in human white blood cells of certain substances from insects to defend it when invaded by immune blood cells is increased. We experiment with changes in the total number of blood cells through the blood cells which increases and decreases, as well as to observe whether the immune response through any route is to evaluate what happens. Hemocyte population was analyzed in the last instar larvae of Spodoptera exigua. Granulocyte and plasmatocyte were predominant (>75%) types of hemocytes, whereas spherulocyte, prohemocyte, and oenocytoid hemocytes were observed in small densities (5~10%). Total hemocyte counts (THCs) were varied among different ages (day1-day5) of the last instar, in which day 3 larvae (L5D3) had the maximal density. Upon bacterial challenge to L5D3 larvae, THC was further enhanced within 2 h and then decreased to background level. This rapid THC increase in response to bacterial challenge was inhibited by injection with dexamethasone (1 ㎍ per larva). However, the addition of arachidonic acid reversed the inhibitory activity of dexamethasone and allowed the larvae to increase THC. This THC increase was mediated by cyclooxygenase products, but not by lipoxygenase products.