Chromosomal level of Korean Diadegma fenestrale (Jeju strain, JK-2023a) of genome assembly was achieved through a combined approach utilizing Nanopore long-read sequencing and Illumina NovaSeq short-read sequencing (approximately 217.2× coverage). The assembled genome spans 221.1 Mb, comprises 68 scaffolds, with most of the genome contained within 11 chromosomal level scaffolds. The completeness of the assembly is reflected in BUSCO assessment, with values reaching 99.6%. Scaffold N50 was 17.4 Mb, and GC % was 40%. RNAseq was performed using RNA extracted from larvae, pupae, and adults at various developmental stages (trimmed RNA-Seq data, 11.3 Gb), and a total of 13,544 genes were predicted by synthesizing the transcriptome information with the annotation information of five closely related species such as, Campoletis sonorensis (GCA_013761285.1), Venturia canescens (GCF_019457755.1), and Nasonia vitripennis (GCF_000002325.3, and GCF_009193385.2). Of these, 13,498 genes were identified by BLAST and are being further analyzed. Although the frequency of DfIV genome integration into the host’s 11 chromosomes varies from 0 to 32%, it was confirmed that all 62 DfIV genome fragments were inserted into the Hymenopteran host genome.

Chromosomal level genome draft was assembled using a Korean Diadegma fenestrale (Jeju strain, KJ09). This assembly was achieved through a combined approach utilizing Nanopore long-read sequencing (approximately 134X coverage) and Illumina NovaSeq short-read sequencing (approximately 83X coverage). The assembled genome spans 243 Mb, comprises 160 scaffolds, with most of the genome contained within 11 chromosomal level scaffolds. The completeness of the assembly is reflected in BUSCO assessment, with values reaching 99.6%. Gene annotation is not completed. A symbiotic virus, Diadegma fenestrale Ichnovirus (DfIV) genome revealed 62 non-overlapping circular DNA segments the aggregate genome size was approximately 240 kb. Although the frequency of DfIV genome integration into the host’s 11 chromosomes varies from 0 to 32%, it was confirmed that all 62 DfIV genome fragments were inserted into the host genome. A total of 123 ORFs were predicted from the DfIV genome and most of those were expressed in the host’s ovary. This result may be contradictory to existing theories, but we propose a new hypothesis that some genes possessed by viruses may play different roles depending on the type and state of the host. Additionally, this phenomenon can be considered in relation to coevolution with the hosts.

The genus Diadegma is a well-known parasitoid group and some are known to have symbiotic virus so called polydnavirus (PDV, more specifically Ichnovirus, IV). The presence of the IV in the Diadegma species a has already been identified more than a decade ago. Previously we reported a DfIV, 62 genomic segments, 247kb from D. fenestrale which parasitized wide range of lepidopteran species (Generalist). However, DsIV from D. semiclausum which parasitized in the P. xylostella (Specialist) has 48 genomic segments, 208kb. Finally, 123 ORFs were re-annotated (repeat element protein, 41; cysteine motif protein, 11; viral innexin, 6; viral ankyrin, 8; polar residue rich protein, 7; N gene, 3; Neuromodulin protein, 2 and not assigned gene, 45). DsIV also have most of lepidopteran immunosuppression gene families and 103 ORFs annotated (repeat element protein, 36; cysteine motif protein, 8; viral innexin, 7; viral ankyrin, 6; polar residue rich protein, 7; N gene, 3; and not assigned gene, 36). Certainly, although viral species specific segments exists, two IVs showed high similarity in most of segments. However DfIV have some more number of that genes such as cysteine motif protein and viral ankyrin. Two Diadegma species also showed difference in mitochondrial genome structure. Therefore we concluded that two species has their own evolutionary linage depending on the lepidopteran hosts with its own symbiotic virus.

The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus. A novel IV was discovered from the calyx of D. fenestrale female. This virus was named as D. fenestrale Ichnovirus (DfIV) and its genome contains 62 circular DNA segments with an aggregate size of approx. 240kb. We examined its gene expression patterns depending on the hosts (hymenopteran host, D. fenestrale and lepidopteran host, Plutella xylostella) and stages of P. xylostella based on RNAseq. Among 62 genome segments, only two segments such C15 and D14 were highly expressed in ovary of D. fenestrale. On the other hands, five segments were differently expressed depending on the developmental stages (3 rd and 4 th instar larvae). Additionally we confirmed integration of DfIV genome segments into the chromosome from the Plutella xylostella via genome sequencing of the lepidopteran host.

감자뿔나방살이자루맵시벌(Diadegma fenestrale, 벌목>맵시벌과>자루맵시벌 아과)은 감자뿔나방(Phthorimaea operculella), 배추좀나방(Plutella xylostella) 등 의 유충 내부기생성 천적으로서, 국내에서는 2009년 제주도 대정읍의 감자밭에서 처음 채집, 보고된 기생봉이다. 이 천적 기생봉을 실내사육하면서 사육조건에 따라 감자뿔나방 유충에 대한 기생률 및 암수 출현율이 각각 다른 양상을 보였다. 설탕 10% 용액과 물을 각각 급여한 기생봉에 감자뿔나방 유충 20마리를 각각 노출시킨 결과, 설탕액 급여구에서 평균 6.6마리의 기생봉이 발생하여 물 급여구 3.8마리, 무 급여구 3.6마리보다 많았다. 천적 기생봉 및 감자뿔나방 성충으로 출현하지 못하고 유충 상태에서 치사한 개체수는 무급여 8±2.1, 물급여 6.4±1.9, 설탕급여 7.4±1.3 마리였다. 따라서 기생봉 출현수와 유충 치사수를 합산한 실질적 천적 방제효과는 설탕 > 무 > 물 급여구 순으로 나타났다. 부화 7일차 감자뿔나방 유충을 기생봉에 24 시간 노출시킨 후 10, 15, 20, 25, 3 0℃ 생육상에 넣고 기생봉 성충이 출현하는 기간을 조사하였다. 모든 온도 조건에 서 기생봉 암컷의 우화기간은 수컷보다 길었다. 고온 조건에서 감자뿔나방의 우화 기간은 상대적으로 기생봉보다 짧았다. 즉 온도가 상승할수록 감자뿔나방 성충이 기생봉 성충보다 먼저 출현하였는데, 이는 생물적 방제 측면에서 감자뿔나방이 먼 저 출현하여 기주식물에 산란하는 것이 천적의 기생활동에 유리하게 작용할 것으 로 생각된다.

The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus, PDV. A novel IV was discovered from the calyx of D. fenestrale female. This virus was named as D. fenestrale Ichnovirus (DfIV). The encapsidated DfIV genome contains 65 circular DNA segments with an aggregate size of 247,191 bp. Based on BLAST analysis, a total of 120 ORFs were predicted as follows: rep; 48, cys-motif; 11, vinnexin; 10, vankyrin; 9, PRRP; 3 and other unassigned genes (39). These viral genes were expressed in lepidopteran hosts (Phthorimaea operculella and Plutella xylostella) after parasitization which means DfIV genome segments were integrated into lepidopteran hosts. This study was focused on this result. Based on gene expression profile, candidate promoter and integration motifs were selected and then, fused with eGFP as a reporter gene. Modified DfIV genome segment was ligated to a commercial containing f1 ori and Ampr gene to propagate in E. coli. We have named this fusion vector as pIN. The construction methodology of pIN and its application would be further discussed in detail.

The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus, PDV. A novel IV was discovered from the calyx of D. fenestrale female. D. fenestrale has more than two hosts, including PTM and DBM. The oviposition and survival rate results showed that D. fenestrale preferred PTM to DBM as hosts. Nevertheless, the developmental period and morphology of D. fenestrale were not significantly different between PTM and DBM. To identify these phenomena, DfIV genome expression patterens were compared between PTM and DBM under various conditions. DfIV genes were more widely expressed in PTM than in DBM after parasitized by D. fenestrale, particularly at the initial point. In addition, large numbers of DfIV genes were expressed only in PTM and they showed differential expression patterns between two lepidopteran hosts. This DfIV genome expression plasticity showed a dependency on the lepidopteran host species and parasitization time, suggesting that it may contribute to the parasitoid survival rate increase. This may be one of the key elements that determine the symbiotic relationship between PDV and parasitoid.

The physiology of parasitic wasp control of their lepidopteran hosts' not only includes injecting their egg but also various factors such as symbiotic virus. This study was focused on the investigation of sophisticated interaction between parasitoid (Diadegma fenestrale) and their host (Plutella xylostella) in P. xylostella larva at transcriptome level, to check whether it is parasitized or not. Short-read deep sequencing method (Hiseq2000) was used for the transcriptome analysis. De novo assembly of cDNA sequence data generated 196,081 contigs between 201bp and 15,853bp in length. Some detoxification enzymes such as cytochrome P450 and Immune-related genes such as antimicrobial peptides were up-regulated after parasitism. Expression of symbiotic ichnovirus genes was detected in parasitized larvae with 55 contigs identified from five ichnovirus gene families including vankyrin, viral innexin, repeat elements, a cysteine-rich motif, and polar residue rich protein. This investigation provides a detailed information on differential expression of P. xylostella larval genes and symbiotic ichnovirus genes following parasitization.

The Diadegma fenestrale is known as parasitoid on potato tuber moth, Phthorimaea operculella and diamondback moth, Plutella xylostella. The Diadegma genus is reported to have symbiotic virus, ichnovirus, D. fenestrale Ichnovirus (DfIV) was identified from this species which is a first report. DfIV showed typical ichnovirus shape with two membranes surrounding the virus capsid. To identify DfIV genes, whole genome sequencing based on GS-FLX was conducted using purified total DfIV genomic DNA extracted from D. fenestrale calyx. About sixty ORFs were analyzed and several typical ichnovirus gene families were detected such as cys-motif, repeat element, vinnexin and vankyrin. Present study was focused on the gene expression patterns in two different lepidopteran hosts.

Potato tuber moth (PTM), Phthorimaea operculella (Zeller), is a serious pest of potato (Solanum tuberosum L.) plants in the fields from warm temperate to tropical climates, and tubers stored under ambient temperatures post-harvest. In recent years, concerns on PTM occurrence and damage to potato are increasing due to climate change resulted in global warming. To search potent natural enemies of PTM, we collected >150 PTM larvae from potato fields in Jeju-do in May, 2009 and then reared in the room until wasp adults emerging, which was identified by Lee J.W. as Diadegma fenestrale (tentative Korean name, 감자뿔나방맵시벌). This is the first report in Korea. When supplied 10% sugar solution on adults to rear, the longevity was 12.2±2.0 day, which was much longer than supplied with only water (5.0±1.4 day) or none (3.1±1.1 day). When the 2nd larvae of PTM, 10 days after hatch, dwelled in potato tubers were supplied to parasitoid adults for parasitization, the developmental periods from egg oviposition to adult emergence was 24.0±2.9 day, and the sex ratio of newly emerged adults was 0.27. The parasitization ratio indoor showed a kaleidoscopic characteristics, ranged from 30.0% to 71.4%. It represented that many factors might be related for the successful parasitization to PTM larvae.

The Diadegma fenestrale was known as parasitoid on potato tuber moth, Phthorimaea operculella and diamondback moth, Plutella xylostella. This species, genus Diadegma are first reported from Korea. DfIV showed typical ichnovirus shape which two membranes surround virus capsids. The genome contents of DfIV consist about sixteen double-stranded DNA segments ranging 2 to 6 kb. To identify DfIV genes, whole genome sequencing based on GS-FLX was conducted using purified total viral DNA extracted from D. fenestrale calyx. About sixty ORFs were analyzed and several typical polydna virus gene family detected such as cys-motif, rep, vinnexin and vankyrin. This is the first report of DfIV and these lepidopteran host immune suppression genes will be deeply identified.