Insects impact human health through vector-borne diseases and cause major economic loses through damaging crops and stored agricultural products. Insect-specific growth regulators (IGR) represent attractive control agents because of their safety to the environment and humans. Here, we report identification of plant compounds that are antagonists of the insect-specific juvenile hormone (PJHANs), using the yeast two hybrid system transformed with the mosquito JH receptor as a reporter assay. We show that these compounds act by inhibiting larval growth and reproduction in mosquitoes. We also demonstrate that PJHANs affect the JH receptor, Methoprene-tolerant (Met), by disrupting its complex with CYCLE, formation of which is required for mediating JH action. We isolated five diterpene secondary metabolites with JH antagonist activity from two plants, Lindera erythrocarpa and Solidago serotina. They are effective in causing mortality of mosquito larvae at relatively low LD50 values. Two of these diterprenes affect Met function, leading to reduction in expression of Met target genes and causing retardation of follicle development in mosquito ovaries. Developing potent compounds counteracting JHaction (JH antagonists) would find a wider range of control applications. However, so far such JH antagonists have not been developed. Here, we report the discovery of potent JH antagonists in plants, which represents an innate resistance mechanism of plants against insect herbivores. These newly discovered plant JH antagonist compounds could be used as the starting material for developing novel insecticides.

The Asian honeybee, Apis cerana, is a native honeybee species in Korea which is important in agriculture for pollination and honey production. For better understanding of the physiology of A. cerana, high-throughput Illumina transcriptome sequencing was performed to analyze the gene expression profiles of queen, worker and larva. A total of 219,799,682 clean reads corresponding to 22.2Gb of nucleotide sequences was obtained from the whole body total RNA samples. The Apis mellifera reference mRNA sequence database was used to measure the gene expression level with Bowtie2 and eXpress software and the Illumina short reads were mapped to 11,459 out of 11,736 A. mellifera reference genes. Total of 9,221 genes with FPKM value greater than 5 of each sample group were subjected to evolutionary genealogy of genes: Non-supervised Orthologous Groups (eggNOG) with BLASTX for gene ontology analysis. The differential gene expression between queen and worker, and worker and larva were analyzed to screen the overexpressed genes in each sample group. in the queen and worker sample group, total of 1,766 genes were differentially expressed with 887 and 879 genes overexpressed over two folds in queen and worker, respectively. In the worker and larva sample group, total of 1,410 genes were differentially expressed with 1,009 and 401 genes overexpressed over two folds in worker and larva, respectively.

Sacbrood virus (SBV) is one of the most fatal pathogens against Asian honeybee, Apis cerana. This virus cause failure of the insect larvae to pupate and death of the adult insects. This study has analyzed the host genes affected by viral infection, by comparing the expression level of host transcripts infected with or without SBV. As a first step, we sequenced the cDNA libraries of Asian honeybee by using illumina RNA sequencing. The sequences were de novo assembled to acquire honeybee transcriptome sequences. The transcriptome was annotated by the sequence comparison to known protein sequences by BLASTX and evolutionary genealogy of genes: Non-supervised Orthologous Groups (eggNOG) database with functional categories and description. By mapping the RNA-seq data to de novo assembled transcripts, we characterized the differentially expressed transcripts between SBV-infected and non-infected Asian honeybee.

Melanization is a unique defence mechanism in arthropods involved in wound healing and pathogen encapsulation. Phenoloxidases (PPOs) are key enzymes of melanization, which mediate the enzymatic conversion of tyrosine to eumelanin. A serine-protease (SP) cascade, similar to the blood-clotting cascade of vertebrates, proteolytically activates prophenoloxidases to phenoloxidases. This proteolytic activation is tightly controlled by serpins and other melanization inhibitors. Melanization has been regarded as one of key immune responses against malaria parasites in mosquitoes. The ookinete melanization of both the simian malaria parasite, Plasmodium cynomolgi, and of the rodent parasite, Plasmodium berghei, prevent parasite development in the human malaria vector, An, gambiae. However, the recent studies revealed a melanization response regulated by Serpin-2 and two C-type lectins (CTL4 and CTLMA-2) was shown to result in ookinete melanization but did not affect the development of the natural human parasite P. falciparum in the mosquito. Instead of melanization, TEP1/APL1/LRIM1 complement-like pathway has been identified as major immune response that regulate parasite loads in the natural association of An. gambiae and P. falciparum. The studies by me and my colleagues revealed another melanization response independent on Serpin-2. Genome analyses of mosquitoes revealed a large expansion of the PPO, SP, and serpin genes potentially involved in the melanization pathway. This expansion was devoted to existence of at least two distinct SP-Serpin regulation modules in controlling separate melanization responses, tissue and hemolymph melanization, in the mosquito, Aedes aegypti. Tissue melanization regulated by Serpin-2 has role in melanotic tumor formation, but not in ookinete melanization. Hemolymph melanization regulated by Serpin-1 and a couple of SPs was activated by the infection of various pathogens and is involved in anti-malarial defense against the avian malaria parasite, P. gallinaceum. A new type of regulator, CLSP2, negatively modulate this hemolymph melanization. Cross-talk between hemolymph melanization and complement-like pathway will be discussed.

우리나라 화장품의 살균⋅보존제 성분은 포지티브 리스트로 관리되고 있다. 포지티브 리스트는 적절한 정량분석법이 필요하지만 아직 분석법 개발이 미비한 상황이다. 본 실험에서는 가스크로마토그래피와 불꽃이 온화검출기(GC/FID)를 이용하여 살균⋅보존제 성분 14종을 동시분석 하는 방법을 개발하였다. 분석법의 validation 결과 특이성을 확인하였고, 검량선의 직선성은 dehydroacetic acid (0.9891)를 제외한 14종에서 상관계수가 0.9997 이상으로 양호하였다. 검출한계(LOD)와 검량한계(LOQ)는 각각 0.0001 mg/mL ∼ 0.0039 mg/mL와 0.0003 mg/mL ∼ 0.0118 mg/mL로 나타났으나, dehydroacetic acid는 각각 0.0204 mg/mL, 0.0617 mg/mL로 나타났다. 반복성은 1.0% 이하로 나왔으나 dehydroacetic acid는 7.1%로 나왔다. 회수률은 96.9% ∼ 109.2% 나타났다. 본 실험방법으로 유통 중인 화장품 50건을 검사한 결과 화장품에 주로 사용되는 살균⋅보존제는 chlorophene, phenoxyethanol, benzyl alcohol, parabens 이고, 모두 배합한도 이내로 검출되었다.