A total of nine Korean indigenous goats were used in a cross-over arrangement to give nine replicates per treatment, and they were housed individually assigned to 1 of 9 dietary treatments. Nine treatments were 0, 500, and 1000 ppm of quercetin supplementation in diets by mixing roughage and concentrate with different ratios (RC ratio) of 3:7 (RC 30), 5:5 (RC 50) and 7:3 (RC 70). Nutrient utilizations of dry matter, crude fat and NDF were not affected by neither RC ratio nor dietary quercetin (p>0.05), but the rate of crude protein and ADF increased in animals in RC 70 group regardless of quercetin supplementation (p<0.05). In addition, higher RC ratio increased (p<0.05) N retention and N retention rate. Total VFA, acetic acid, propionic acid, iso-butyric acid, butyric acid, iso-valeric acid and valeric acid contents were not affected (p>0.05) by dietary quercetin. Meanwhile, lower total cholesterol level exhibited in animals in RC 70 group compared to RC 30 or 50 groups, unrelated to dietary quercetin (p<0.05), however other plasma parameters were not influenced (p>0.05) by RC ratio and dietary quercetin. Our results indicated that both RC ratio and dietary quercetin may not directly affect the production indices and immune responses in Korean indigenous goat

As basic research to develop HEV and EV agricultural field machinery, the present study analyzes the technical trend of electric agricultural field machinery through product analysis, paper analysis, and patent analysis concerning HEV and EV in the automobile, construction machinery, and agricultural machinery sectors. For product analysis, the homepages of companies in these sectors were consulted to analyze the number of products of each company. For paper analysis, key words related to HEV and EV were selected, a search formula was drawn up, and articles search sites were consulted. And for patent analysis too, key words were selected and then a search formula was drawn up to examine published patent applications or registered patent applications, and trends were analyzed by structure, country, and year. The number of HEV and EV products were 17 in the automobile area, 8 in construction machinery, and 4 in agricultural machinery. Notably, in the agricultural machinery area, all HEV and EV products were from advanced companies overseas. In terms of papers, papers published in the past 5 years were searched and 33,195 papers were from the automobile area, 3,806 were from construction machinery, and 2,687, the fewest papers, were from the agricultural machinery area. A search of patents in the electric drive technology area in Korea, USA, and Japan, and Europe showed 1,927 valid patents, with 1,120 in Japan, 497 in USA, 193 in Korea, and 117 in Europe. Analysis of the trend of research on electric agricultural field machinery by product, paper, and patent shows the development of HEV and EV technology in Korea is insufficient compared to USA, Japan, and Europe, which means rapid technological development is needed.

Among hemipteran insects which is the most important insect vector of plant viruses, small brown planthopper, Laodelphax striatellus, transmits the rice stripe virus (RSV) causing rice stripe disease. For effective control of RSV, it is important to understand interaction between RSV and L. striatellus. Therefore, in this study, expressed sequence tag (EST) databases were generated based on 454 GS-FLX pyrosequencing for comparative transcriptome analysis between nonviruliferous and RSV-viruliferous L. striatellus. By comparing the two EST libraries, we showed that 108 host genes were significantly up-regulated and 28 host genes were significantly down-regulated in viruliferous insects. Interestingly, genes encoding ribosomal proteins were mainly up-regulated in viruliferous L. striatellus, whereas genes related to translation were concentrated in the downregulated cohort. These RSV-dependently regulated genes may have important function in the behavior of planthopper or the transmission of RSV.

Bacillus thuringiensis serovar mogi of a novel serogroup (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 only megaplasmid (> 30 MDa) on which the toxin genes were occasionally located. Sequence analysis using 454-pyrosequencing revealed that the megaplasmid harbored at least seven putative cry genes, showing about 84%, 75%, 73%, 58%, 84%, 39% and 75% similarities in amino acid sequences with Cry27Aa, Cry19Ba, Cry20-like, Cry56Aa, Cry39ORF2, Cry8Ba and Cry40ORF2, respectively. These cry genes were cloned to the Escherichia coli-B. thuringiensis shuttle vector, pHT1K, 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 real-time PCR (qrtPCR) from the wild type strain as well as transformant strains. The 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.

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.

In the transcriptome surveys of Laodelphax striatellus, several cDNA sequences showed a high level of similarities to the insect picorna-like virus genomes. Interestingly, there was no sequence similarity between picorna-like virus sequences from the RSV-viruliferous and those from the non-viruliferous L. striatellus. Picorna-like virus from the non-viruliferous L. striatellus was a geographical isolate of Himetobi P virus (HiPV). The genome of the HiPV was 9,272 nt in length excluding the poly(A) tail and contained two open reading frames (ORFs), which were separated by a 176 nt intergenic region that functions as an internal ribosome entry site (IRES). The 5' ORF encodes the non-structural proteins and the 3' ORF encodes the capsid proteins. The partial genomic RNA of the picorna-like virus from the RSV-viruliferous L. striatellus, LsPV-2, was 8,769 nt in length excluding the poly(A) tail and contained a single, large open reading frame (nt 1–8,535) encoding a 2,845 aa polyprotein. In terms of sequence similarity, identity, and genome organization, LsPV-2 resembled insect picornalike viruses belonging to the family Iflaviridae. A phylogenetic analysis based on RNA-dependent RNA polymerase (RdRp) sequence showed that LsPV-2 was most closely related to the deformed wing virus (DWV). The HiPV and LsPV-2 were incompatible each other in L. striatellus, suggesting that these two picorna-like viruses may have important functions in transmission of the RSV.

Plasmids from Bacillus thuringiensis have been implicated in pathogenicity as they carry the genes responsible for different types of diseases in mammals and insects. B. thuringiensis serovar mogi of a novel serogroup (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 only megaplasmid (> 30 MDa) on which the toxin genes were occasionally located. Sequence analysis using 454-pyrosequencing revealed that the megaplasmid harbored at least seven putative cry genes, showing about 84%, 75%, 73%, 58%, 84%, 39% and 75% similarities in amino acid sequences with Cry27Aa, Cry19Ba, Cry20-like, Cry56Aa, Cry39ORF2, Cry8Ba and Cry40ORF2, respectively. These cry genes were cloned to the Escherichia coli-B. thuringiensis shuttle vector, pHT1K, and then introduced into an acrystalliferous B. thuringiensis Cry-B strain for further molecular characterization. This novel 3a3b3d type strain, B. thuringiensis serovar mogi, could be used as a good resource for studying unknown mosquitocidal cry genes.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved baculovirus gene whose homologs are found in all lepidoteran Group I NPV, but its function is unknown so far. To determine the role of ac11 in baculovirus life cycle, ac11 knock-out mutant, Ac11KO, was constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that ac11 transcript was first detected at 6 h post-infection (p.i.) and accumulated to maximum at 48 h p.i., indicating that ac11 is belong to late gene. When the genomic DNA of Ac11KO was transfected into Sf9 cells, viral replication was restricted to a cell transfected originally. While viral transmission of the Ac11KO was not observed in Sf9 cells, production of budded virus (BV) in Sf9 cells transfected with Ac11KO was observed by transmission electron microscopy (TEM). These results suggest that the ac11 is essential for AcMNPV to produce infective BV.

Rice stripe virus (RSV), the type member of the genus Tenuivirus, causes rice stripe disease and the viral transmission is mediated through the sucking by small brown planthopper, Laodelphax striatellus. Considerations have been mainly focused on the protection of rice from RSV and/or the planthopper, rather than the interaction between RSV and the insect. To clarify the interaction, in this work, mRNA was extracted from RSV-viruliferous planthopper with non-viruliferous control, and expressed sequence tag (EST) databases were generated based on 454 GS-FLX pyrosequencing technology for comparative analysis. RSV-viruliferous planthopper had ca. 2500 isotigs, which included genes on biological process (19%), cellular component (13%), molecular function (22%) and no hits (46%) from gene ontology (GO) analysis; this structure was similar to the control. However, in the viruliferous planthopper, 109 isotigs were up-regulated and 660 isotigs were down-regulated, compared to the non-viruliferous control. These RSV-dependently regulated genes may have important function in the behavior of planthopper or the transmission of RSV.

Plasmids from Bacillus thuringiensis have been implicated in pathogenicity as they carry the genes responsible for different types of diseases that in mammals and insects. A novel serogroup (H3a3b3d), B. thuringiensis strain K4 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 strain K4 (designated as serovar mogi) had only one large plasmid (>200kb) on which the toxin genes were occasionally located. A 454 pyrosequencing was used for the complete sequencing of the large plasmid. The sequence analysis showed that k4 plasmid had at least seven putative cry genes, ending up to showing 84%, 75%, 73%, 58%, 84%, 39% and 75% homology with Cry27Aa, Cry19Ba, Cry20-like, Cry56Aa, Cry39ORF2, Cry8Ba and Cry40ORF2 toxins in amino acids, respectively. This novel 3a3b3d type strain, B. thuringiensis serovar mogi, can be used as a good resource for studying unknown mosquitocidal cry genes. The E. coli-B. thuringiensis shuttle vector, pHT1K was used to clone these cry genes for characterization. In each clone, the level of transcription and production of crystal proteins will be investigated in near the future.

Among 154 putative ORFs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), ac78 and ac79 are highly conserved genes in baculovirus, but their functions in the virus life cycle have been unknown so far. To determine their roles in AcMNPV replication, knockout mutants, ac78KO and ac79KO, were constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that both of ac78 and ac79 transcripts were first detected at 6 hours post-infection, and accumulated to maximum at 24 hours post-infection, suggesting that both of ac78 and ac79 are belong to late gene. When the genomic DNA of ac78KO was transfected into Sf9 cells, viral replication was restricted to a single cell infection. These results demonstrated that the ac78 play an important role in BV production, and therefore is essential for AcMNPV to mount a successful infection. Whereas Sf9 cells infected with the ac79KO showed normal viral symptoms such as rounding and swelling, OBs were not observed from majority of infected cells. These results suggested that the ac79 might play an important role in OB production.

Experiments were carried out to develop an optimal nutrient solution for the single-stemmed rose (Rosa hybrida L.) 'Red velvet' in a closed aeroponic system. Plants were grown in 1/3, 1/2, 1, or 3/2 strength of the nutrient solution of National Horticultural Research Station in Japan (NHRS). Significantly less changes of pH and EC (dS·m-1) in the drainage were observed in 1/2 strength treatment as compared to other treatments. The NO3-N, K, Ca, and Mg concentrations in the drainage solution of 1/2 strength treatment were maintained at optimal levels. These results indicated that the rose uptakes of both nutrients and water was more stable than those in other concentration. The concentration of macronutrients in nutrient solution were adjusted based on the ratio of nutrient:water (n/w) taken up by plants grown in the 1/2 strength solution. The composition of the new solution (classified the University of Seoul (UOS) solution) was as follow; NO3-N 8.8, NH4-N 0.67, P 2.0, K 4.8, Ca 4.0, Mg 2.0 me·L-1. To further evaluate new solution on crop growth, the rose 'Red Velvet' was grown again in l/2, 1, and 2 strength UOS solution to compare with 1.0 strength PBG (proefstion voor bloemisterij en glasgroenpe) solution. Overall the plant growth, including the stem length and number of five-leaflet leaves was higher in 1.0 strength of UOS solution than other treatments. Results presented in this study indicate that the nutrients in the UOS solution are well balanced for the single-stemmed rose in the closed aeroponic system.

Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. We constructed a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis and to enable a better understanding of the evolution of leguminous species. The draft genome sequence covers 80% of the estimated genome, of which 50.1% consists of repetitive sequences. In total, 22,427 high confidence protein-coding genes were predicted. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (Vigna reflexo-pilosa var. glabra) provided genomic evidence of a recent allopolyploid event. To further study speciation, we compared de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max and Cajanus cajan. The species tree was constructed by a Bayesian Markov chain Monte Carlo method using highly confident orthologs shared by all 24 accessions. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.

Leaf structure is one of the important agronomic traits. A rolled leaf mutant was induced from an ethyl methane sulfonate (EMS)-treated japonica rice, 'Koshihikari'. The rolled leaf mutant showed phenotypes of reduced leaf width and leaf rolling. In addition, several abnormal morphological characteristics were observed, including dwarfism, defected panicle, delayed germination, and lower seed-setting. Microscopic analysis revealed that the number of small veins was decreased and the sizes of adaxial bulliform cells were reduced in the mutant leaves. The genetic study with two F2 populations from the crosses of the rolled leaf mutant with 'Koshihikari' and Milyang23 suggested that the mutant phenotype might be controlled by a single dominant gene.

The mutants of sugary-2 (su-2), floury (flo), shrunken-1 (shr-1), and dull-1 (du-1) were crossed to waxy (wx) to produce F2 seeds. Chi-square analysis on the segregating ratio of the F2 seeds revealed that flo, su-2, and shr-1 were independently transmitted with wx, while wx was epistatic over du-1. The floury and sugary-2 were crossed to Hwasunchalbyeo, a waxy variety, and then the F4 of floury-waxy and sugary-2-waxy seeds were developed, respectively. As the parents phenotypes of sugary-2 and floury, the grains of these two lines showed lower hardness and grain weight than normal grain of Hwacheongbyeo. For alkali digestive value (AVD), the sugary-2-waxy showed lower ADV than Hwacheongbyeo. For the gel consistency of grain flours, the floury was medium like Hwacheongbyeo, while those of the sugary-2, floury-waxy, and sugary-2-waxy were soft like Hwasunchalbyeo. The amylose contents in the grains of the sugary-2 and floury were decreased to ~15% whereas that of Hwacheongbyeo was 19.1%. All the lines showing waxy endosperm (Hwasunchalbyeo, floury-waxy, and sugary-2-waxy) showed less than 4% amylose contents. Interestingly, the free sugar content in the brown rice was increased to 9.27% in the sugary-2-waxy, showing transgressive segregation phenomenon where the free sugar contents in its parents, sugary-2 and Hwasunchalbyeo, were 5.98% and 3.98% respectively. Also, the floury-waxy showed transgressive segregation phenomenon, containing 6.15% of free sugar content in the grains