A new fumigant, carbonyl sulfide (COS), has potential for use as a replacement for methyl bromide, yet its mechanism of toxicity to insects remains poorly understood. In this study, transcriptome analysis was performed on Tribolium castaneum malpighian tubules and fat bodies, which are known to play an essential role in energy storage and utilization in insect species. In total, upon exposure to COS, 3,034 and 2,973 genes were differentially expressed in the T. castaneum malpighian tubules and fat body, respectively. These differentially expressed genes comprise a significant number of detoxification-related genes, including 105 P450s, 18 glutathione S-transferases (GSTs), 82 ABC transporters, 25 UDP-glucosyltransferases and 42 carboxylesterases and mitochondrial–related genes, including 9 complex Ⅰ genes, 2 complex Ⅱ genes, 1 complex Ⅲ gene, 9 complex IV genes, 8 complex V genes from both malpighian tubules and fat body tissues. Moreover, KEGG analysis demonstrated that the upregulated genes were enriched in xenobiotic metabolism by ABC transporters and drug metabolism by other enzymes. We also investigated the role of carbonic anhydrases (CAs) in toxicity of COS using dsRNA treatment in T. castaneum. These results show that CA genes have a key role in toxicity of the COS. Furthermore, the results of transcriptomic analysis provide new insights into the insecticidal mechanism of COS fumigation against T. castaneum and eventually contribute to the management of this important stored grain pests.

Salt stress is one of the most limiting factors that reduce plant growth, development and yield. However, identification of salt-inducible genes is an initial step for understanding the adaptive response of plants to salt stress. In this study, we used an annealing control primer (ACP) based GeneFishing technique to identify differentially expressed genes (DEGs) in Italian ryegrass seedlings under salt stress. Ten-day-old seedlings were exposed to 100 mM NaCl for 6 h. Using 60 ACPs, a total 8 up-regulated genes were identified and sequenced. We identified several promising genes encoding alpha-glactosidase b, light harvesting chlorophyll a/b binding protein, metallothionein-like protein 3B-like, translation factor SUI, translation initiation factor eIF1, glyceraldehyde-3-phosphate dehydrogenase 2 and elongation factor 1-alpha. These genes were mostly involved in plant development, signaling, ROS detoxification and salt acclimation. However, this study provides new molecular information of several genes to understand the salt stress response. These genes would be useful for the enhancement of salt stress tolerance in plants.

The small brown planthopper (SBPH), Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus (RSV) which often causes significant reduction of yield in the field. Buprofezine is an effective insect growth regular (IGR) pesticide to control planthoppers, however, since the use of buprofezine for more than a decade, it has caused a certain resistance of SBPH. To survey the responses of SBPH to buprofezine, we exposed 4th instar SBPH to 200 ppm buprofezine by dipping method, and extracted total RNA for RNA-seq by Illumina platform. The quality filtered raw reads of cDNA obtained from experimental and control SBPH were subjected to Bowtie2 followed by eXpress computer program to compare the differential gene expression which will be important information for pest control methods using RNAi.

Insect Growth Regulators (IGRs) are insecticides that disrupt the normal development of target insects by inducing symptoms such as premature molting or supernumerary larval stages. Juvenile hormone systems become the targets of two types of IGRs: the Juvenile Hormone Agonists (JHAs) and Juvenile Hormone Antagonists (JHANs). Pyriproxyfen is one of the chemical compounds widely used as JHA to control many kinds of insects while Kanakugiol is a plant-extracted compound which acts as JHAN. The small brown planthopper, Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus which often causes significant reduction of yield in the field. In order to analyze the differential gene expressions of L. striatellus upon JHA and JHAN treatment by using next generation sequencing technique, we sprayed Pyriproxyfen and Kanakugiol on 4th instar nymphs of L. striatellus respectively, and extracted total RNA for RNA-seq. The quality-filtered Illumina sequence reads of the control, JHA, and JHAN treated samples were mapped to the reference gene sequences by using the Bowtie2 software. Then the results of mapping by Bowtie2 were analyzed by eXpress software to quantity the differential gene expression.

To determine differential gene expression profiles in the salivary gland of a water stick-insect, Ranatra chinensis Mayrt, a subtractive cDNA library was constructed by suppression subtractive hybridization. The salivary gland was determined among three salivary gland-like tissues by investigating transcription levels of five trypsin genes isolated from R. chinensis. The major transcripts encoding trypsins (64.4% of the total ESTs) were eliminated from the library and then remaining salivary gland-specific genes were searched. A total of 643 expressed sequence tags (ESTs) were clustered and assembled into 148 contigs (49 multiple sequences and 99 singletons), among which 35 contigs had matched BLASTx hits (E ≤ 1.00E-4). Salivary apyrase occupied 5.6% (36 ESTs) of the library. Apyrase is known to be released by female mosquitoes or blood-sucking assassin bugs to prevent blood clots during blood sucking. Therefore, apyrase in the salivary of R. chinensis might allow R. chinensis to facilitate feeding. Several contigs encoding acid phosphatase, hyaluronidase, prophenoloxidase, and dipeptidylpeptidase IV, commonly found in venoms of Hymenoptera, were also identified from the salivary gland-specific library. Discovery of salivary glandspecific genes should promote further studies on biologically active components in the saliva of R. chinensis.

To determine differential gene expression profiles in the salivary gland of a predatory flower bug species, Orius laevigatus Fieber, a subtractive cDNA library was constructed by suppression subtractive hybridization. The major transcripts encoding trypsins (28.6% of the total ESTs) were eliminated from the library and then remaining salivary gland-specific genes were searched. A total of 513 expressed sequence tags (ESTs) were clustered and assembled into 129 contigs (64 multiple sequences and 65 singletons). About 58% were matched with insect genes. In total, 38 genes (179 ESTs) were found from the library by BLASTx search. A hemolysin-like protein occupied ca. 8% (41 ESTs) of the library. Hemolysin is known to destruct cells including blood cells by forming pores on the cell membrane. A hemolysin-like salivary protein of O. laevigatus might be hemolytic against the prey cells, thereby allowing O. laevigatus to facilitate feeding. Several contigs encoding lipase was also identified from the salivary gland-specific library. Discovery of salivary gland-specific genes should promote further studies on biologically active components in the saliva of O. laevigatus.

To determine differential gene expression profiles in the venom gland and sac (gland/sac) of a solitary hunting wasp species, Eumenes pomiformis Fabricius (1781), a subtractive cDNA library was constructed by suppression subtractive hybridization. A total of 541 expressed sequence tags (EST) were clustered and assembled into 102 contigs. In total, 37 genes were found from the library by BLASTx search and manual analysis. A eumenitin-like venom peptide, EpVP1, occupied ca. 26% of the library. A novel venom peptide, EpDTX, shared sequence similarity with trypsin inhibitors and dendrotoxin-like venom peptides known as K+ channel blockers, implying it could be responsible for the paralysis of prey. As well as phospholipase A2 and hyaluronidase known to be main components of wasp venoms, several contigs encoding enzymes, including metalloendopeptidases and a decarboxylase likely involved in the processing and activation of venomous proteins, peptides, and neurotransmitters, were also isolated from the library. The presence of a gene encoding insulin-like growth factor binding protein suggests that solitary hunting wasps might control the prey to stay in larval stage by their venom. The abundance of these venom components in the venom gland/sac and alimentary canal was confirmed by quantitative real-time PCR.

To determine differential gene expression profiles in a cypermethrin-resistant strain (CR) of diamondback moth, Plutella xylostella Linnaeus (1758), a subtractive cDNA library was constructed by suppression subtractive hybridization. A total of 1196 expressed sequence tags (EST) were clustered and assembled into 579 contigs (100 multiplets and 479 singletons). About 46% (267) of 579 contigs had the matched BLASTx hits (E ≤ 10-5). Among these, 143 contigs had similarity to proteins with assigned molecular function in the Functional Catalogue database, and most of them (86%) were homologous to the genes from insects, particularly to Lepidoptera (56%). The contigs encoding carboxylesterase and cytochrome P450 known to be involved in the insecticide resistance were found in the library. They were identified as pxest3, pxest4, and CYP9G2 gene by 5' and 3' RACE. Among these, pxest4 was determined to be 2-fold over-transcribed in the CR strain by qrtPCR. Several contigs encoding enzymes including cytochrome oxidase subunit I that are likely involved in the insecticide resistance were also identified from the library. Discovery of the genes specific to cypermethrin resistance should promote further studies on the molecular mechanisms of insecticide resistance in P. xylostella.

Melatonin has several known physiological functions, the main one being synchronization of daily and seasonal rhythms. In addition, melatonin has been reported to influence reproduction and behavioral rhythms with varying results depending on the species. To date, it remains unknown how this rhythm in locomotor activity is controlled endogenously, although there must be coordination of chemical and molecular drivers. However, the species is poorly characterized at molecular level with little sequence information available in public databases. The aim of study was to clarify involvement of endogenous melatonin rhythms and locomotor activity in day-night activity of the eel, Anguilla japonica which is an economically important but endangered species. The levels during daytime (zeitgeber time; ZT 6) were significantly (P<0.05) lower than those during nighttime (ZT 18). A similar pattern was persisted under DD conditions, whereas it disappeared under LL conditions and ocular melatonin levels remained low. Therefore, it is likely that ocular melatonin levels of the nocturnal eel reared under LD and DD conditions fluctuate in a daily/circadian manner and night-related physiological processes are dependent on eel locomotor activities which is a nocturnal species. We found that similar number of genes were differentially expressed between day (ZT6) and nighttime (ZT18), suggesting that during the nighttime also important in differential gene expression with daytime. This work also provides essential information for further studies investigating the molecular basis of daily/circadian system in this species.

Carotenoid isomerase (CRTISO) catalyzes the isomerization of prolycopene to all-trans-lycopene in the carotenoid biosynthetic pathway. We isolated a full-length promoter region of CuCRTISO from Citrus unshiu. We determined if the promoter encoded organ-specific or developmental-specific expression, and identified possible cis-acting promoter elements. The full-length promoter and two truncated versions were fused to the β-glucuronidase (GUS) gene and transformed into Arabidopsis thaliana. Transgenic lines expressing the full-length promoter (pCiso-Prom1) and truncated promoters (pCiso-Prom2 and pCiso-Prom3) showed the same developmental and organ-specific activity. GUS expression was detected in the cotyledon and root at 5 and 10 days after germination, mature leaf, and anther. The CuCRTISO promoter contained several cis-acting elements involved in hormonal and environmental stress. Drought stress or abscisic acid treatment did not induce GUS expression in any transgenic lines. Heat stress induced GUS expression in the pCiso-Prom1 line; this promoter construct contains the heat-stress responsive element (HSE). Ethylene and cold-stress treatments induced GUS expression only in the pCiso-Prom3 line, although all transgenic lines contained the same cis-acting ethylene and low-temperature response elements. which could indicate the existence of unknown repressor element(s) in the CuCRTISO promoter. These studies indicate that CuCRTISO promoter activity is regulated in a developmental and organ-specific manner that responds to heat, cold, and ethylene. These results provide new insights into the role of cis-acting element(s) in CuCRTISO promoter activity. (This research was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2010-0007627 and 2009-0094059), and by Golden Seed Project, Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), Rural Development Administration (RDA) and Korea Forest Service (KFS), Republic of Korea)

Soybean is an important crop with useful traits such as the high seed protein and oil contents. Soybean reproduction is sensitive to temperature over 35℃. To obtain database of gene expression profiles, we used soybean cultivars, sensitive and tolerant. RNA sequencing was performed to find differentially expressed genes in two Korean soybean cultivars under heat stress condition. The transcriptomic changes in each cultivar under heat stress. We found 2727 common transcripts in two soybean cultivars under heat stress, and selected 20 transcripts to heat stress response genes. The 20 selected genes were analysed using BLAST2GO and PLANEX. The genes were major factor in co-expression networks. It appears that these 20 genes were mainly attributable to heat stress.

Transition of the resting primordial follicle to the growing primary follicle is a critical process for female reproduction, but its mechanism is poorly understood. The present study was conducted to investigate gene expression profile at the primordial-primary follicle transition process. We isolated total RNA of female mouse ovary at day1 (contains only primordial follicles) and day5 (contains primordial and primary follicles) and synthesized cDNA using annealing control primers (ACP; Seegene, Inc., Seoul, Korea). ACP provides annealing specificity and sensitivity to the template and allows to identify only authentic differentially expressed genes (DEGs). We used total 80 ACPs for PCR, observed PCR products on 2% agarose gel, cloned 42 DEGs using TOPO TA cloning vector, sequenced, and analyzed by BLAST search. Sequences of 34 clones significantly matched database entries while 4 clones were novel and 4 clones were EST. Two of 34 genes were specifically expressed only in day 5 ovaries (Sui1-rs1, Apg3p/Aut1p-like), and rest of 32 genes were expressed in both stages but were differential in amount. Differential expression was confirmed using semiquantitative RT-PCR, and there was no false positive. Anx11 and Pepp2-pending were highly expressed genes in day1-, while BPOZ, Ches1, Kcmf1, NHE3, Nid2, Ninj1, SENP3 and Survivin were highly expressed genes in day5-ovary. List of genes would provide insight for further study of mechanism regulating primordial-primary follicle transition.

성장을 멈추고 있는 원시난포(primordial follicle)에서 난포발달이 개시되어 1차난포(primary follicle)로 발달하는 조절기전은 잘 알려져 있지 않다. 이 초기 난포발달 과정에 관여하는 유전자를 알아내기 위해 suppression subtractive hybridization(SSH)을 사용하였다. 생후 1일과 5일째의 생쥐 난소로부터 얻은 cDNA로 forward와 reverse subtraction을 수행하여 각각 day1과

DNA 회복효소인 MPG는 DNA의 퓨린기에 결합되어 있는 메틸기 등 이물질을 염기와 함께 제거하는 작용을 한다. 본 연구에서는 노던 블롯팅 방법을 이용하여 Balb/c mice의 각 조직별로 발생단계별 mRNA 발현 정도를 조사하였다. 뇌와 콩팥조직에서는 출생직후에 발현이 가장 활발하였으며, 성체시기까지 비교적 높은 활성도가 유지되었다. 위장 조직에서는 출생직후에서 일주일 후까지는 명확히 관찰되었으나, 그 이후는 발현이 약화되었다. 간장과 폐조직에서는