The potato tuber moth (PTM) is an oligophagous pest on solanaceous crops. We tested adult olfactory behavior and larval development on 5 cultivated tomato varieties leaves including Moneymaker, Campari, Ailsa craig, LA3475 and E6203, and one wild species, S. pimpinellifolium. Bioassay on PTM larvae showed highest developmental performances in Ailsa craig, and lowest in LA3475. Tomato leaf hydro-distilled oils of Moneymaker, Campari, Ailsa craig, S. pimpinellifolium and E6203 showed strong attractiveness and LA3475 exhibited repulsiveness for mated adult PTM of both sexes in two-armed bioassay. The hydro-distilled oils of all tomato leaves contain the fatty acid n-hexadecanoic acid as a major compound. We conformed presence of n-hexandecanoic acid (palmitic acid) in the leaves by GC-MS. LA3475 consists of (1-hexyltetraecyl) cyclohexane (10.5%) as the second major compound which was absent in all other tomato varieties. A standard sample palmitic acid was tested against the pest at 0.1, 1 and 10 mg/ml did not show any significant olfactometer effect. On other hand, phytol showed a weak attractiveness (60.7-63.6%) and (57.6-60.5%) for male and female PTM, respectively at 0.1, 1 and 10 mg/ml. Additionally, SEM of these tomato varieties leaves showed they have trichomes with distinct shapes. We are working on HPLC, and GC-MS analysis of specific chemicals in trichomes of these tomato varieties leaves.
Insect cuticle tanning (pigmentation and sclerotization) is a complex and vital process, which includes hydroxylation of initial amino acid, tyrosine, to DOPA and decarboxylation of DOPA to dopamine. In the pigmentation process, dopamine further undergoes two N-acylation reactions to yield N-acetyldopamine (NADA) and N-β-alanyldopamine (NBAD). In the former reaction, arylalkylamine N-acetyltransferase (AANAT1) converts dopamine to NADA, and in the later reaction, aspartate 1-decarboxylase (ADC) provides β-alanine, which is conjugated with dopamine catalyzed by NBAD synthase (Ebony) for production of NBAD.
In this study, we performed functional genomics of TmAANAT1, TmADC and Tmebony to determine whether they are required for cuticle pigmentation in Tenebrio molitor adults. Loss of function of these genes by RNAi caused the significantly darker body color than that of control animals. Note that, although all phenotypes exhibited dark cuticle pigmentation, RNAi of either TmADC or Tmebony only altered brownish outer region of the cuticle to dark/black. In contrast, RNAi of TmAANAT1 had no effect on the brown hue of the outer cuticle layer, but less or no pigmented inner region of the cuticle became significantly darker than those of control adults. These results suggest that, like that seen in TcAANAT1- or TcADC-deficient Tribolium castaneum adults, NADA produced by a reaction by TmAANAT1 contributes the lighter inner cuticle layer(s), whereas NBAD appears to do the highly pigmented outer cuticle layer(s) of the cuticle of T. molitor adults. This work was supported by NRFs (NRF-2015R1A6A3A04060323 and NRF-2018R1A2B6005106).
Lately, CRISPR-Cas9 has become one of the most essential tools to understand gene’s function. In the honeybee, however, the application of CRISPR technology has been hindered by various factors leading to very few reports of success in genome editing. Among these, collection of honeybee embryos for microinjection has been a time-consuming procedure, mainly limiting the applicability of the genome editing technique to honeybees. To improve the drawbacks of the conventional plastic plug-based system, we have developed a film-assisted honeybee embryo collection system (FECS) using transparent film as a detachable bottom layer. In this new system, eggs are laid on the detachable film surface and collected in a batch, and thus no additional alignment is required for microinjectoin. As the film can be easily replaced with in a few seconds, embryo collection can be repeated continuously after a single caging of a queen. Also, unlike conventional plug-based systems, the new system utilizes 100% of the eggs laid by the queen, thereby increases the yield three times in theory. The main unit of the system can be printed with ordinary SLA/DLP type 3D printer and the stl file for 3D printing will be distributed online.
The honey bee soluble acetylcholinesterase 1 (AmAChE1) is overexpressed under the overwintering and brood rearing-suppressed conditions. To investigate the role of AmAChE1 in regulating acetylcholine (ACh) titer, ACh concentrations both in the head (neuronal) and abdomen (non-neuronal) were analyzed. ACh titer was significantly lower in both tissues of worker bees under the overwintering and brood rearing-suppressed conditions compared to control bees. The expression levels of another two factors that regulate ACh titer, choline acetyltransferase (AmAChT) and acetylcholinesterase 2 (AmAChE2), were not altered as judged by qPCR and native PAGE, suggesting that the lower ACh titer was mainly regulated by AmAChE1. For precise verification of AmAChE1 as an ACh titer regulator, honey bees were put under brood rearing-suppressed condition to induce AmAChE1 and injected AmAChE1 dsRNA to knock down the gene. The ACh titer of AmAChE1-knocked down honey bees was 1.9 and 2.6 folds higher than that of control bees in head and abdomen, respectively. Taken together, in spite of its extremely low catalytic activity, the overexpression of AmAChE1 is likely to be related with the low level of ACh homeostasis, perhaps via ACh sequestration, under brood rearingsuppressed condition, and likely induce metabolic changes through ACh receptors-related pathways.
The pea aphid, Acyrthosiphon pisum, requires the nutritional endosymbiont, Buchnera, for the production of essential amino acids. However, it is unclear if host plant diet that varies in essential amino acids influences aphid regulation of its nutritional symbioses. We hypothesized that aphid genes respond to host plant diet when aphids feed on their specialized (alfalfa) compared to universal host plant diet (fava), which vary in essential amino acid concentrations. Using RNA-Seq and whole genome bisulfite sequencing, we compared the gene expression profiles and DNA methylation distributions of specialized aphid cells that harbor Buchnera (bacteriocytes) when aphids feed on their specialized compared to their universal host plant diets. Our results show that bacteriocyte transcription and methylation patterns differ between host plant diets. When aphids feed on their specialized host plant, they significantly up-regulate and/or hypo-methylate key aphid genes in bacteriocytes related to the amino acid metabolism, including glutamine synthetase (GS) and the glutamine transporter ApGLNT1. Moreover, regardless of which host plant aphids feed on, we observed significant up-regulation and differential methylation of the key genes in the amino acid metabolism and the glycine/serine metabolism in aphid bacteriocytes. We suggest that the regulatory response of key symbiosis genes in bacteriocytes allows aphids to feed on a specialized host plant diet with suboptimal nitrogen concentrations.
Beauveria comprises entomopathogenic fungi frequently isolated from insect cadavers, among which Beauveria bassiana is the most widely studied species of entomopathogenic fungal genus for its high potential as a biological pesticide. Even though it has been reported that B. bassiana is a heterogeneous assemblage of strains, little is known about the factors that might drive the genetic diversity among various isolates. In this work, we ought to study the gene diversity of 33 isolates in order to figure out the relationship between their gene diversity and biological features. First, we analyzed gene sequences that are involved in developmental and immunological processes of the fungal isolates and host. Now we are checking whether the gene diversity is related to the genes function, the isolates geographic localization, thermo-tolerance and virulence of the isolates. The obtained results so far showed no correlation between the genetic diversity and the fungal geographic localization. But interestingly, a positive correlation has been obtained between the virulence and the genetic diversity based on the internal transcribed spacers (ITS). Further analyses are aiming to confirm the obtained data and to analyze possible correlations between the genetic diversity and morphological and transcriptional data of the studied isolates. Based on the obtained results, we could much strongly focus on interesting genes which might be related to improving or screening highly virulent fungal agents as biological control agents.
To identify the venom components and their expression patterns of some Aculeata bees/wasps, venom gland-specific transcriptome analysis was conducted. FPKM values were normalized with the average of the transcription level of reference gene (a-tubulin). Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, the information on venom compositions of solitary wasps obtained in this study was not sufficient to generalizae this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insuline-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more abundant in social wasp venom. In the venom gland trancsriptome of bumblebees, major allergens or pain producing factors were barely identified, implying that bumblebee venoms are relatively less toxic than those of social or solitary wasps.
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.
Ticks are obligatory hematophagous ectoparasites, which can transmit various pathogens including bacteria, protozoa, and viruses via salivary secretion during feeding. Understanding salivation in ticks is crucial for the development of novel methods to prevent tick-borne disease transmission. The central nervous system (synganglion) of ticks controls salivary glands via several neuropeptidergic innervation: myoinhibitory peptide (MIP), SIFamide, and elevenin. These neuropeptides are thought to be modulators of dopamine’s action controlling the salivary glands including inward fluid transport into the lumen of salivary glands acini and emptying lumen solute into salivary duct by pumping and gating. These actions are via two distinct dopamine receptors, dopamine receptor (D1) and invertebrate D1-like dopamine receptor (InvD1L), respectively.
The domestic natural enemy industry was formed by public institutions before the natural enemy companies were formed. In 1995, during the decentralization period, the technololgy of natural enemy breeding developed by the Rural Development Administration and the Agricultural Research Institute of each province were spread to the Agricultural Technology Centers of each county, Agricultural Technology Centers has supplied a large number of natural enemies to farmers for free. Since the beginning of the 2000s, when the first natural enemy company called Korea-IPM was created in Korea, the natural enemy industry boomed with the birth of Cecil Co., Ltd., a large natural enemy company.
Prior to the birth of a natural enemy company, Domestic methods of using nautral enemies to release the natural enemies against the target pests and to test their effectiveness were similar to those of using chemicals. After then, the introduction of banker plant, a natural enemy protection plant, began to develop the concept of natural enemy protection limited to Aphidius colemani, and the effect was excellent. However, there were many cases where the success and failure of biological control were staggered because only the use value of the natural enemy for the target pest was dealt with.
Therefore, in our laboratory, we are exploring and developing of the using method of trap plant that can concentrate densities by attracting insect pests. Also, when using a variety of natural enemies in a single plant, we are studying strategies to increase the pest control efficiency by identifying the interraction between natural enemies and determining their amount of releasing.
The continuous emergence of chemical-mediated residual toxicity and insect resistance have enforced the regulation of synthetic pesticides. Synthetic pesticides with novel mode of actions could be developed to overcome these issues, but as an alternative biopesticides with more efficacious control activity could be developed by the advanced technology. In pest management, entomopathogenic fungi have high potential in reducing pest population to an economic threshold, and some of isolates have been commercialized. However still a novel application strategy needs to be considered for successful industrialization. An insight on fungal genes in whole genome and transcriptome levels is necessary to understand the role of genes in pathogenesis and genetic diversity of fungal genes. Herein, we have identified the genetic differences of entomopathogenic fungi using whole genome sequencing of Beauveria bassiana (Bb) and tried to understand the interaction between fungus and insect using RNA-seq. We have obtained the whole genome sequence of Bb JEF-007 using PacBio sequencing technology and compared the transcriptomes of Bb JEF-007 and bean bug, Riptortus pedestris before and after the fungal infection using Hiseq 2000 system.
토양해충의 포식성 천적인 뿌리이리응애[Stratiolaelapsscimitus(Wo mersley)]를 이중박스 시스템을 활용하여 대량생산할 수 있는 방법을 개발하였다. 사육상자는 외부(22L, 54×36×18 cm3)및 내부상자(6.5L, 31×22×11 cm3)로 구성된 플라스틱상자이다. 내부상자에는 뿌리이리응애의 은신처 역할을 하는 왕겨(4L)를 담았다. 외부상자에는 물을 1L 채우고 그 위에 내부상자를 넣고 외부상자 덮개를 덮었다. 이 시스템은 포식성응애의 탈출을 막고 은신처의 습도를 유지할 수 있다. 천적의 먹이원으로서 긴털가루응애[Tyrophagusputrescentiae(Sch rank)]를 쌀겨에 사육하였으며, 뿌리이리응애와 마찬가지로 2중상자에서 증식시켰다. 천적응애상자에 주 1회 먹이응애를 제공하면서 25±1℃, 상대습도 60±10% 조건에서 사육하였다. 뿌리이리응애의 밀도는 4주에 3배로 증가하였으며 최종적으로 약 12만마리/상자(4L)였다. 이 방법은 간단한 뿌리이리응애 대량사육기술로서 토양해충의 생물적 방제에 큰 도움이 될 것으로 판단한다.
미국선녀벌레(Metcalfa pruinosa)는 외래 침입해충으로 조사가 시작된 2009년 이후 발생지역과 발생면적이 계속 증가하여 2018년에는 123개 시군구의 21,154ha에서 발생이 보고되고 있다. 미국선녀벌레는 농경지와 그 주변 산림지에서 활동하기 때문에 정부와 지자체간 협업을 통해 농림지 동시방제를 5월하순경(부화약충 대상)과 7월 중순경(성충 대상) 2회 실시하고 있다. 농촌진흥청에서는 미국선녀벌레의 기생천적인 선녀벌레집게벌(Neodryinus typhlocybae)의 안정적인 도입과 미국선녀벌레에 대한 생물적 방제원으로서 국내 정착을 위해 이탈리아 파도바대학과 국제협력사업을 2017년부터 수행하고 있다. 선녀벌레집게벌은 완주에서 월동이 가능하였으며 6월 초에 50%이상 우화를 하였다. 2018년도에 온실(완주)과 야외(수원, 태안)에서 증식하여 1,000여 개체 이상을 확보하였으며 도입천적의 안정적인 국내 정착과 확산 및 활용 방안에 대해 논의하고자 한다.
2010년대 들어 화학물 기반의 작물보호제에대한 환경독성 및 잔류 문제의 대두, 병해충에 대한 저항성 증대와 이에 따른 신규 작물보호제 개발 비용의 증가 등은 바이오 작물보호제에 대한 관심을 증가시키고 있다. 특히 Bayer의 AgraQuest 인수, BASF의 Becker Underwood인수, 신젠타의Pasteuria Bioscience 인수나 Monsanto와 Novozyme의 BioAg., FMC와 Chr. Hansen 등의 기술 및 제품개발 협력관계 구축 등은 바이오 작물보호제에 대한 다국적 작물보호제 기업의 높은 관심을 보여주는 것이다. 본 발표에서는 미생물을 활용한 글로벌 바이오 작물보호제 개발 동향 및 팜한농에서 개발한 바이오 작물보호제 개발 사례를 소개하고 향후 바이오 작물보호제 산업화를 위해 요구되는 사항들을 정리하여 논의하고자 한다.
천적을 이용한 친환경농법은 이미 그 실효성이 인정되어 농업 선진국을 중심으로 유용하게 이용되고 있는 검증된 농법임에도 불구하고 한국내에서 일부 대형 농가를 제외하고는 널리 보급되지 못한 것이 현실이다. 이러한 이유는 한국 농업이 가지고 있는 물리, 환경적 요인과 더불어 기존 작물보호제의 사용패턴에 익숙해져 있는 소농들의 관행이 가장 큰 이유로 분석된다. 이러한 현실을 감안하여 보다 현실적인 한국형 천적방제 모델 개발이 다각도에서 진행중이다. 국내 기후, 환경에 맞는 토착천적의 개발을 비롯하여, 해충의 방제적기를 놓치지 않도록 하는 서식처 보존 전략등의 개발이 진행중이며 여기에 이러한 내용을 간략히 소개하고자 한다. 이러한 현지 친화형 천적방제 모델의 개발은 차후 천적농법 이용을 촉진시키는데 있어서 많은 도움을 줄 것으로 보인다.
Since fireflies are considered to be insects that come from green or clean regions, many local festivals are held around the country. Fireflies in Muju, Chonbuk, Mt. Gyeyang in Incheon, Anter Village in Chungbuk, and O-chang Chungbuk (Chungbuk Agricultural Research Extension Services) are the most popular festivals. In the case of O-chang, Chungbuk, we looked at the possibility of fireflies succeeding as a local festival. The O-chang Firefly Festival was held from June 1, 2018 to June 5, 2018, marking the ninth anniversary of the Firefly Festival as the topic of “Firefly·Insect industry Festival.” Firefly (Luciola lateralis, the larvae are aquatic) is a main exhibiting insect. About 30 living species of aquatic insects, attracting insects, agricultural insects, and edible insects, and insect photo artwork were displayed. The event was organized not only watch, but also experience for making various insect models, such as cooking insects and a exhibition of illustrated poems of insect subjects. In addition, they performed special stage performances such as bubble shows, magic shows, puppet shows, and insect movies to watch while waiting. As a result, the number of visitors increased 67 percent year-on-year to 21,700 this year. It is considered a success by relieving visitors of various free attractions and experiences.
땅강아지는 농작물 해충이지만 약용, 낚시미끼용, 체험학습용으로 활용되는 산업곤충이다. 땅강아지의 실내사육 및 대량증식에 관한 연구는 2010년부터 시작되었으며 최근 곤충산업에 대한 관심이 높아지면서 곤충사육농가와 차별화된 곤충소재를 찾는 농민들에 의해 사육이 시도되고 있다. 하지만 대량사육에 대한 경험부족 및 안정적인 판매처 확보에 대한 불안함으로 산업화로의 연결이 미흡한 실정이다. 왕지네는 가장 잘 알려진 약용절지동물로서 중국의 대량사육기술을 습득한 국내 사육농가와 곤충관련 연구기관에서 꾸준한 사육이 시도되었으나 대부분 누대사육에 실패하여왔다. 그동안의 실패사례와 시행착오를 바탕으로 국내 환경에서 성공가능성을 높일 수 있는 왕지네 대량 누대사육방법의 모색이 필요하다. 또한 땅강아지와 왕지네의 약리활성물질 분석을 통한 효능 및 기능성에 대한 다양한 연구를 통해 산업화가 가능할 것이다.
벼메뚜기는 분류학적으로 메뚜기목(Orthoptera), 메뚜기과(Acrididae)에 속하는 곤충으로 작물에 피해를 주는 해충으로 인식하고 있지만 예로부터 단백질 보충을 위하여 채집하여 튀기거나 볶음요리 로 이용해 왔다. 벼메뚜기는 갈색거저리, 쌍별귀뚜라미 등과 함께 식품공전에 등록되어 있으나 1년 1세대로 가을철에 채집하여 이용하는 실정으로 공급확대에 한계가 있다. 최근 우리나라에서 벼메뚜기를 사육하고자 하는 농가와 다양한 용도로 가공을 통해 이용하려는 수요가 점차 증가하고 있으나 연중사육기술이 개발되지 않아 대량사육하는 농가가 거의 없는 실정이다. 이에 벼메뚜기의 연중대량사육기술 뿐만 아니라 인공사료 및 자동화 사육시설 개발에 따른 생산비 절감 기술 개발이 필요한 실정이다. 벼메뚜기의 먹이로는 여름작물로 옥수수, 수단그래스, 겨울작물로 밀, 보리 등의 볏과작물이 있으며 많은 양의 생엽과 발육에 적합한 작물의 선발 및 볏과작물의 통곡실 가루를 이용한 인공사료의 개발로 먹이공급 부족 시 15~20일 급여 가능하며, 벼메뚜기 사육시설로 단독형, 연결형, 절충형 등의 사육시설을 개발하여 연결형 사육시설의 수확량과 생존율이 가장 높았으며, 예취 급이 노동력이 단독형의 16% 소요되었다.
2003년 부산에서 처음 발견된 외래종인 등검은말벌(Vespa velutina)은 2018년 현재 전국적으로 확산되어 양봉을 포기하는 농가가 속출할 정도로 심각한 경제적 피해를 초래하고 있다. 말벌류의 방제는 유인 포획기를 사용하는 것이 가장 효과적인 방법으로 알려져 있다. 유인 포획기는 당 성분의 섭식 유인제에 1차 유인 포획된 말벌이 알람페로몬을 발산하면서 2차 유인을 여기하여 포획력을 극대화시키는 원리이다. 그러나 등검은말벌은 개체군 성장을 위해 단백질원이 필요하기 때문에 당성분의 휘발성 물질에 의존하는 유인제를 탑재한 기존 포획기만으로는 충분한 등검은말벌 개체 수 감소 및 방제에 한계가 있다. 본 연구에서는 이와 같은 문제점을 해결하여 등검은말벌 포획율을 향상시키고 말벌 피해로 인한 꿀벌 개체군 보호를 위하여 꿀벌통 일체형의 포획시스템을 개발하였다. 꿀벌 사냥 후 말벌집으로 회귀하는 습성, 최초 꿀벌 포획에 성공한 양봉장으로 회귀 후 지속적으로 사냥하는 습성, 어두운 곳에 있을 때 밝은 쪽을 선호하는 습성 등 다양한 생태 특성을 반영하여 시스템을 고안되었다. 즉, 최초 사냥성공 시 등검은말벌을 포획함으로 동일 개체에 의한 반복적 피해를 방지하는 새로운 방식의 포획법이다. 본 연구를 통해 개발된 포획시스템은 양봉 농가의 가장 큰 피해 요인인 말벌 사냥에 의한 꿀벌 개체수 감소를 방지함으로서 양봉 산업의 유지 및 발전에 기여할 것으로 기대된다.
Insect antimicrobial peptides (AMPs) have been characterized more than 150 peptides since identification of cecropin in the hemolymph of pupae from Hyalophora cecropia in 1980. Therefore, it is considered that insects are good species of AMPs selection. Insect AMPs are small (below 10 kDa), cationic, and amphipathic with variable length, sequence, and structure. They perform a critical role on humoral immunity in the insect innate immune system against invading pathogens such as bacteria, fungi, parasites, and viruses. Most of insect AMPs are induced rapidly in the fat bodies and other specific tissues of insects after septic injury or immune challenge. Then the AMPs subsequently released into the hemolymph to act against microorganisms. These peptides have a broad antimicrobial spectrum against various microorganisms including anticancer activities. Insect AMPs can be divided into four families based on their structures and sequences. That is α-helical peptides, cysteine-rich peptides, proline-rich peptides, and glycine-rich peptides/proteins. For instance, cecropins, insect defensins, proline-rich peptides, and attacins are common insect AMPs, but gloverins and moricins have been identified only in lepidopteran species. In this presentation, we focus on AMPs from insects and discuss current knowledge and recent progresses with potential application of insect AMPs.