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.

Rice stripe virus (RSV) is one of the serious plant pathogenic viruses for rice and mediated by small brown planthopper, Laodalphax striatellus. So far, the studies have been mainly focused on the interaction between the host plant and the virus. In this study, for better comprehension of the interactions among Rice stripe virus, rice and small brown planthopper, transcriptomes of the RSV-viruliferous (RVLS) and non-viruliferous L. striatellus (NVLS) were comparatively analysed. For this, non-viruliferous L. striatellus were collected from non-infected rice field and fed RSV-infected rice for 5 days. With the RNAs prepared from the RSV-viruliferous and the non-viruliferous small brown planthoppers, we conducted Illumina RNA sequencing (Hiseq 2000) and then two transcriptome databases were generated from RVLS and NVLS, respectively. The transcriptome of RVLS and NVLS were campared to figure out how the gene expression of the insects affected by Rice Stripe Virus. RSV-dependently regulated genes analysed from this study may have important functions in the transmission and replication of RSV.

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.

The novel serogroup of Bacillus thuringiensis serovar mogi (H3a3b3d) was isolated from fallen leaves, sampled in a forest region of the city of Mungyeong, Korea. Plasmids from B. thuringiensis have been implicated in pathogenicity as they carry the genes responsible for different types of diseases in mammals and insects. In this study, the genome sequence of the strain was determined. The 6.0-Mb genome of B. thuringiensis mogi contains three replicons: a circular chromosome (5.40-Mb) encoding 5,652 predicted open reading frames (ORFs), and two megaplasmids, pMOGI364 (364 564 bp) and pMOGI222 (222 348 bp). The G+C contents of these replicons ranged from 31.3% to 34.2% for pMOGI364 and pMOGI222, respectively. There are six putative cry genes, cry19Bb1, cry73Aa, cry20Bb1, cry27Ab1, cry4Aa and cry56Ba1, distributed on these two megaplasmids. To investigate the role of these genes in crystal production, the expression profiles of these toxin genes were analyzed by quantitative PCR (qPCR) from the wild type strain. Also, these cry genes were cloned to the Escherichia coli-B. thuringiensis shuttle vector, pHT1K under the control of its own promoter and then introduced into an acrystalliferous B. thuringiensis Cry-B strain for further molecular characterization.

Rice stripe virus disease (RSVD), one of the most serious disease of rice is mediated through the sucking by small brown planthopper, Laodalphax striatellus. So far, the studies have been mainly focused on the interaction between the host plant and the virus. In this study, for better comprehension of the interactions among the host plant, vector insect and plant-pathogenic virus, we investigated transcriptome of the vector insect and the differences between viruliferous and naïve L.striatellus. For this, naïve L. striatellus were collected from non-infected rice field and 50 L.striatellus of them were fed RSV-infected rice for 5 days. With the RSV-viruliferous and the naïve insects, we conducted Illumina RNA sequencing (Hiseq 2000) and obtained 175,243,488 and 146,031,348 reads from viruliferous and naïve L.striatellus, respectively. These reads were assembled into contigs and two transcriptome databases were generated. The transcriptome of naïve and RSV-viruliferous L. striatellus were campared to figure out up-regulated or down-regulated genes. These RSV-dependently regulated genes may have important function in the behavior of planthoppers or the transmission of RSV.

Proteinaceous insecticidal proteins, Cry proteins, from Bacillus thuringiensis (Bt) are insecticidal proteins that are highly active against several species of Lepidoptera. Thus, cry genes encoding these Cry proteins have been widely applied for construction of transgenic crops resistant to pest insects. In this study, through the 3D structure prediction and accompanying mutagenesis study for the Mod-Cry1Ac, 7 and 16 amino acid residues from domain I and II, respectively, responsible for its insecticidal activity against larvae of Spodoptera exigua and Ostrinia furnacalis were identified. We used site-directed mutagenesis to improve the insecticidal activity of Mod-Cry1Ac, resulted 31 mutant cry genes. These mutant cry genes encodes potent insecticidal proteins in the form of crystalline protoxins of 95 kDa. SDS-PAGE analysis of the recombinant polyhedra revealed that expressed Cry proteins was occluded into polyhedra and activated stably to 65 kDa by trypsin. When the insecticidal activities of these mutant Cry proteins against to larvae of P. xylostella, S. exigua and O. furnacalis were assayed, they showed higher or similar insecticidal activity compared to those of Cry1Ac and Cry1C. Especially, Mutant-N16 is considered to have the potential for the efficacious biological insecticide since it showed the highest insecticidal activity.

Plasmids are crucial for determining the pathogenicity and host range of organisms of the Bacillus thuringiensis strains. In this research, a novel serogroup of B. thuringiensis serovar mogi (H3a3b3d), which showed mosquitocidal activity against Anopheles sinensis and Culex pipiens pallens, was isolated from fallen leaves in Mungyeong city, Republic of Korea. In contrast to the complicated plasmid profiles of B. thuringiensis H3 serotype strains, the B. thuringiensis serovar mogi contained two megaplasmids (> 30 MDa) on which the toxin genes were occasionally located. Sequence analysis using 454-pyrosequencing revealed that there are 7 putative cry genes, cry19Bb1, cry73Aa, cry40orf2, cry20Bb1, cry27Ab1, cry56Ba1 and cry39orf2, distributed on the two different megaplasmids, respectively. These cry genes were cloned to the Escherichia coli-B. thuringiensis shuttle vector, pHT1K under the control of its own promoter and p1KSD, which is a recombinant expression vector containing cyt1Aa promoter combined with the STAB-SD sequence, and then introduced into an acrystalliferous B. thuringiensis Cry-B strain for further molecular characterization. To investigate the role of these genes in crystal production, the expression profiles of these toxin genes were analyzed by quantitative PCR (qPCR) from the wild type strain. These results clearly indicate that the cry39orf2 was the dominant ingredient in the crystal. This novel 3a3b3d type strain, B. thuringiensis serovar mogi, could be used as a good resource for studying unknown mosquitocidal cry genes.

ORF78 (ac78) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a baculovirus core gene of unknown function. To determine the role of ac78 in baculovirus life cycle, an ac78-deleted mutant AcMNPV, Ac78KO, was constructed. Quantitative PCR analysis revealed that ac78 is a late gene in the viral life cycle. After transfection into Spodoptera frugiperda cells, Ac78KO produced a single-cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was also confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Analysis of BV and occlusion derived virus (ODV) revealed that AC78 is associated with both forms of the virions and is a structural protein located to viral envelope. Electron microscopy showed that ac78 also plays an important role in embedding of ODV into occlusion body. This study therefore demonstrates that AC78 is a late virion associated protein and is essential for the viral life cycle.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved gene of unknown function. To determine the role of ac11 in baculovirus life cycle, an ac11-knockout mutant AcMNPV, Ac11KO, was constructed. qPCR analysis revealed that ac11 is an early gene in the life cycle. After transfection into Spodoptera frugiperda cells, Ac11KO produced a single cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Electron microscopy showed that ac11 is required for nucleocapsids envelopment to form ODV and their subsequent embedding into OB. This study therefore demonstrates that ac11 is an early gene which is essential for the viral life cycle.

Crystals of proteinaceous insecticidal proteins, Cry proteins, produced by Bacillus thuringiensis (Bt) have been generally used to control insect pests. In this study, through the 3D structure prediction and accompanying mutagenesis study for the Mod-Cry1Ac, 7 and 16 amino acid residues from domain I and II, respectively, responsible for its insecticidal activity against larvae of Spodoptera exigua and Ostrinia furnacalis were identified. To construct novel cry genes with enhanced insecticidal activity, we randomly mutated these 23 amino acid sequences by in vitro muti site-directed mutagenesis, resulting in totally 24 mutant cry genes. For further characterization, these mutant cry genes were expressed as a fusion protein with polyhedrin using baculovirus expression system. SDS-PAGE analysis of the recombinant polyhedra revealed that expressed Cry proteins was occluded into polyhedra and activated stably to 65 kDa by trypsin. When the insecticidal activities of these mutant Cry proteins against to larvae of P. xylostella and S. exigua were assayed, they showed higher or similar insecticidal activity compared to those of Cry1Ac and Cry1C. Especially, among them Mutant-N16 showed the highest insecticidal activity against to both of P. xylostella and S. exigua. Therefore, Mutant-N16 is considered to have the potential for the efficacious biological insecticide.

The baculovirus expression system is one of the most popular methods used for the production of recombinant proteins but has several complex steps which have proved inherently difficult to meet a multi-parellel process. We have developed a novel recombinant bacmid, bEasyBm that enabling easy and fast generation of pure recombinant virus without any purification step. In the bEasyBm, attR recombination sites were introduced to facilitate the generation of recombinant viral genome by in vitro transposition. Moreover, extracellular RNase gene from bacillus amyloliquefaciens, barnase, was expressed under the control of Cotesia plutellae bracovirus early promoter. Therefore, only when the barnase gene was replaced to gene of interest, the bEasyBm could replicate in host insect cells. When the bEasyBm was transposed with pDualBac-EGFP and pDualBac-LUC respectively, there were no non-recombinant backgrounds were detected from unpurified BmEasy-EGFP or BmEasy-LUC stocks. In addition, the resulting recombinant virus, BmEasy-EGFP, showed comparable level of EGFP expression efficiency with the plaque-purified recombinant virus, BmEGFP, which was constructed using bBmGOZA system. Based on these results, high-throughput condition for generation of multiple recombinant viruses in a time was established.

Varieties of Bacillus thuringiensis (Bt) crystal proteins, Cry proteins, have so far been found as one of the most successful biological control agents which are safe to natural environments for a long time. Recently, cry genes encoding these Cry proteins have been widely applied for construction of transgenic crops resistant to pest insects. In this study, through the 3D structure prediction and accompanying mutagenesis study for the Mod-Cry1Ac, 7 and 16 amino acid residues from domain I and II, respectively, responsible for its insecticidal activity against larvae of Spodoptera exigua and Ostrinia furnacalis were identified. To construct novel cry genes with improved insecticidal activity, we randomly mutated these 23 amino acid sequences by in vitro muti site-directed mutagenesis, resulting in totally 24 mutant cry genes. For further characterization, these mutant cry genes were expressed as a fusion protein with polyhedrin using baculovirus expression system. SDS-PAGE analysis of the recombinant polyhedra revealed that expressed Cry proteins was occluded into polyhedra and activated stably to 65 kDa by trypsin. In the further study, we plan to investigate their insecticidal activity against Plutella xylostella, S. exigua and O. furnacalis larvae.

The influenza virus is an important respiratory risk affecting humans, and effective treatments are needed. Some oriental medicines are currently applied for treatment of common colds as well as influenza infection. Previous studies have reported that the therapeutic properties of MA-128 are effective for treatment of psoriasis antiasthmatic and atopic dermatitis. In this study, we investigated the therapeutic properties of the novel herbal medicine, MA-128, for treatment of influenza virus infection by oral administration. MA-128 is an active natural biological compound from herbal-marine origin. The results showed that oral administration of MA-128 in mice could confer a survival benefit against Type A influenza virus infection. Daily oral administration of MA-128 resulted in delayed death in infected mice for three days against mouse adapted H3N2 (A/Philippines/2/82). However, it protected more than 60% of mice from lethal infection of 2009 pandemic H1N1 (A/Korea/CJ01/2009) influenza virus. In addition, lung viral titers were significantly reduced at seven days post infection (~100 times) compared with mock-treated mice and viruses were cleared at 9 dpi only in the MA-128 treated groups. This study demonstrated the potential of the novel herbal medicine, MA-128, as an herbal remedy against influenza A viruses.

꼬마배나무이 (Cacopsylla pyricola)에 저항성 품종육성을 위한 육종재료를 선발코자 월동형 성충수, 단과지내의 산란수, 과총엽내의약충수 및 수관 전체적인 그을음 발생정도를 15개 종 및 종간잡종133개 유전자원을 대상으로 조사하였다. 월동형 성충수는 Pyrus.calleryana 4.6마리, 단과지내 산란수는 P. calleryana 0.3개, 과총엽내약충수는 P. calleryana 0마리, 그리고 그을음 발생정도는 P.betulaefolia, P. calleryana, P. communis, P. hybrid (P.pyrifolia × P.communis), P. lindleyi 0으로 발생 및 피해정도가 가장 낮았으며 종간의 차이가 뚜렷하였다. 전체적으로 대목으로 이용하고 있는 콩배인P. calleryana와 P. betulaefolia가 조사 대상 유전자원 중 가장 높은 항객성(antixenosis)을 보였으나 품질개량에 많은 기간이 요구됨에 따라 실용적으로는 서양배(P. communis) 중 ‘Conference', 'Cascade','Bosc', 'Winter Nelis' 가 가장 낮은 발생 및 피해정도를 보여 꼬마배나무이 저항성 품종 육성을 위한 좋은 육종재료로 판단되었다.

The objective of this study was to determine the effect of macrophages on growth of human colon cancer cells. The results showed that co-culture of colon cancer cells with macrophages inhibited the growth of colon cancer cells (HCT116 and SW620) depending on the number of macrophages, RAW 264.7 cells, and activated THP-1 cells accompanied by down regulation of pSTAT3 in cancer cells. We also found that expression and release of cancer cell growth inhibitory cytokines, IL-1 receptor antagonist (IL-1ra) and IL-10, was increased in macrophages. Blocking of the STAT3 pathway with specific inhibitor and siRNA of STAT3 abolished the growth of colon cancer cells and expression of IL-1ra and IL-10. In addition, neutralization of IL-1ra and IL-10 with antibodies resulted in reversal of macrophage-induced inhibition of cancer cell growth. These data showed that IL-1ra and IL-10 released from macrophages inhibit growth of colon cancer cells through inhibition of the STAT3 pathway.

농촌진흥청 원예특작과학원에서는 2003년에 황색 폼 폰 화형인 ‘Restone’에 자주색 폼폰 화형인 ‘Lollipop’을 교배하여 획득한 실생 집단으로부터 황색 적심 폼폰 화형의 겹꽃 ‘03B1-82’ 계통을 선발하였다. ‘03B1-82’ 계통은 2005년 겨울부터 2007년 3년에 걸쳐 1, 2차 특성검정을 통해 안정성, 균일성 및 흰녹병저항성 검정 및 절화수명에 대해 조사되었고, 2008년에는 우수선발 계통으로 ‘원교B1-142호’ 계통번호를 부여받아 3차 특 성검정을 실시하여 안정성, 균일성에 대한 연차별 재현 성 그리고 주년생산성(촉성, 억제재배), 생산자 및 소 비자 기호성을 평가 받았다. ‘원교B1-142호’는 2008년 에 직무육성품종심의회를 거쳐 ‘옐로우캔디’로 명명되 어졌다. 국화 ‘옐로우캔디’는 10월 하순에서 11월 상 순에 자연 개화하는 절화용 스프레이 추국이다. 황색의 적색화심을 지닌 폼폰화형의 겹꽃으로 초세 및 줄기가 강건하다. 특히 여름철 고온기에도 통상화가 많이 발생 하지 않으며, 촉성 및 억제재배 등 주년생산이 가능하 다. 꽃의 직경이 4.3 cm 내외, 꽃잎수는 소화당 180매 이상, 착화수는 본당 8화 내외의 중형화로 절화수명은 18일 정도이다. 재배상 유의점으로는 둥근모양의 폼폰 형이기 때문에 수확시 꽃이 서로 엉키면 꽃이 떨어지는 경우가 있으며, 여름 장마철 또는 겨울철 환기 부족시 흰녹병 발생을 주의하고 주기적인 방제가 필요하다.

운간초의 분화 품질을 향상시키기 위하여 ‘Kumoma’ 와 ‘Kumoma-Gusa’ 두 품종의 생장과 개화에 미치는 영향을 구명하고자 식물생장억제제 (PGRs) 4종을 처리 하였다. 식물생장억제제 종류와 처리농도는 paclobutrazol (10, 20, 40, 80 mg·L-1), flurprimidol (5, 10, 20, 40mg·L-1), daminozide (500, 1000, 2000, 4000mg·L-1), chlormequat (50, 100, 200, 400 mg·L-1)이고 엽면살포 와 토양관주 방법으로 처리하였다. Paclobutrazol 40 mg·L-1 처리는 운간초의 두 품종에서 초장과 화경 장을 줄이는데 효과적이었다. ‘Kumoma’ 품종에서, paclobutrazol 40 mg·L-1 엽면살포와 토양관주 처리방 법은 초장을 각각 12.6, 12.5 cm로, 화경장은 3.4, 3.3 cm로 줄일 수 있었다. ‘Kumoma-Gusa‘ 품종에서 는 paclobutrazol 토양관주는 처리 농도가 증가함에 따라 초장은 13.2-10.7 cm, 화경장은 3.9-2.0 cm까지 줄일 수 있었지만 꽃의 수는 대조구와 비교했을 때 현 저하게 감소하여서 20개까지 감소하였다. Flurprimidol 20mg·L-1 스프레이와 토양관주 처리는 두 품종 모두 초장과 화경장을 줄일 수 있었다. ‘Kumoma’ 품종에 서 flurprimidol 10 mg·L-1 스프레이 또는 토양관주 처리는 개화 품질을 향상시킬 수 있었고 대조구와 비 교해 꽃의 수는 44.7개 이상, 출하 일수는 5일 정도 빨리 출하 할 수 있었다. ‘Kumoma-Gusa‘ 품종에서 PGRs처리에 의한 초장 과 화경장 억제효과가 가장 큰 것은 paclobutrazol 80mg·L-1과 flurprimidol 40mg·L-1 이었고 paclobutrazol 과 flurprimidol 스프레이와 토양관주 처리에서 각각, 초장은 10.7, 9.9 cm, 화경장은 2.0, 1.5 cm 로 생장 억제효과가 가장 큰 것을 알 수 있었다. 그러나 flurprimidol 40 mg·L-1 토양관주 처리에서 출하일수는 농도에 따라 3-13일 정도 대조구보다 늦어졌고 꽃의 수는 15.6개로 가장 적은 것을 관찰 할 수 있었다. 두 품종 모두에서 daminozide 처리는 초장과 화경장 이 줄어들지 않았지만 ‘Kumoma’ 품종에서는 대조구 와 비교했을 때 꽃의 수가 증가하였다. Chlormequat 처리는 paclobutrazol 과 flurprimidol 처리와 비교했 을 때 화경장 감소효과는 작았고 처리 식물의 잎은 다른 처리에 비해 연한 녹색을 보여주었다. 두 품종 모두에서 chlormequat 과 daminozide 처리는 생육과 개화 품질에 효과적으로 영향을 주지는 않았지만 꽃의 수는 모든 처리농도에서 41개 이상을 보여주었고, ‘Kumoma-Gusa’ 품종에 chlormequat 토양관주 처리는 최대 꽃의 수 63개를 볼 수 있었다. 이번 결과들은 paclobutrazol 과 flurprimidol 처리가 운간초 분화의 품질 향상을 위한 화경장의 생육을 조 절하는 생장억제제로서 사용 될 수 있음을 보여 주었 고, ‘Kumoma’ 품종에서는 paclobutrazol 40 mg·L-1 스프레이는 생육특성을, flurprimidol 10 mg·L-1 토양관 주 방법은 개화관련 특성의 품질을 향상시키는 생장억 제제의 농도로 추천 될 수 있고, ‘Kumoma-Gusa’ 품종 에서는 flurprimidol 20mg·L-1 스프레이와 flurprimidol 5mg·L-1 토양관주 방법이 운간초 분화의 생육특성과 개 화품질을 향상시키는 생장억제제의 농도로 추천 될 수 있다.