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Ying Fang,김우진,이석희,김종훈,하규백,박동환,우라미,박민구,최재영,제연호 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.2
The small brown planthopper, Loadelphax striatellus is one of the serious pest insects of rice plants because it can transmit the rice strip viruswhich often causes significant reduction of yield in the field. Insect growth regulators (IGRs) are insecticides which disrupt the normal development of target insects by inducing symptoms such as premature molting or supernumerary larval stages as the results of hormonal disruption. One of the most important insect hormones, juvenile hormone (JH) becomes the target of two types of IGRs: JH agonists (JHAs) and antagonists (JHANs). Pyriproxyfen is one of the popular IGR insecticideswhich acts as JHA to control many kinds of insects while kanakugiol is a plant-originated chemical which recently revealed its function as JHAN. In order to study the responses of L. striatellus to JHA and JHAN treatments, pyriproxyfen and kanakugiol were sprayed on 4th instar nymphs, and each total RNA samplewas extracted for RNA-Seq by using Illumina sequencing platform. For differential gene expression study, the in silico cDNA library was constructed by Tirinity de novo assembler. The expression levels of each gene were quantified by Bowtie2 and eXpress programs, and then subjected to gene ontology (GO) enrichment analysis to study the changes of gene expression profile. Total of 33 GO terms showed differential expression upon JHA or JHAN treatment, and the genes related to insect hormone biosynthesis were identified. These results will provide important information for future studies of molecular biology and physiology of L. striatellus.
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김우진,이석희,김종훈,Ying Fang,하규백,박동환,최재영,제연호 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.1
The Asian honeybee, Apis cerana, is a native honeybee species in Korea which is important in agriculture for pollination and honey production. Since 2009, A. cerana in Korea has been endangered because of the outbreak of sacbrood virus (SBV) disease which caused serious colony loss. SBV belongs to the genus Iflavirus, and infects the honeybee larvae specifically to cause larval death. For better understanding of the molecular biological aspects of the viral pathogen and insect host, the A. cerana larvae latently infected with SBV virus (AcSBV) and control (AcCtrl) were subjected to high-throughput transcriptome sequencing to compare the gene expression profiles. 101 base paired-end Illumina sequencing generated a total of 211,850,996 and 231,925,174 clean reads from AcSBV and AcCtrl, corresponding to 17.4 Gb and 18.7 Gb of nucleotides, respectively. An in silico cDNA library was constructed by Trinity de novo assembler and TGICL clustering program to make a reference cDNA library containing 67,750 contigs. The quality-filtered Illumina short reads were mapped to the cDNA library, and the top 20,000 of the most frequently mapped contigs by the Bowtie2 program were subjected to the ImmunoDB database to annotate insect innate immune-related genes. The genes with expression levels whichwere two fold higher or lower in AcSBV compared to the AcCtrl, overall a total of 20 contigswere identified and annotated to demonstrate that the latent infection of SBV affects expression pattern of innate immune-related genes.
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김종훈,이수연,최재영,이석희,Ying Fang,하규백,박동환,박민구,우라미,김우진,김주곤,제연호 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.1
Pest resistance of 8 transgenic Bt rice events with a synthetic cry1Ac gene linked to rice rbcS-tp sequence were assessed under laboratory conditions. Bioassays were conducted with neonates and third instar larvae of Cnaphalocrocis medinalis, which is a significant pest of rice in Asia. C. medinalis larvae were shown to be susceptible to all eight events, even though there were differences between the causes of death. The results differed between developmental stages of the larvae, despite the fact that all 8 events led to high mortalities. Neonates displayed feeding avoidance and death by starvation on six Bt rice events. In the case of third instar larvae however, only two events resulted in feeding avoidance. Nevertheless, all Bt rice events were shown to be highly effective against C. medinalis larvae in laboratory conditions. These results may be a significant foundation for the evaluation of improved transgenic Bt rice.
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Improved insecticidal activities of Novel Bacillus thuringiensis Cry1-type genes
Jong Hoon Kim,Song Eun Kim,최재영,유금,이석희,Ying Fang,하규백,박동환,제연호 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.1
Recently, Bacillus thuringiensis (Bt) cry genes encoding insecticidal Cry proteins have beenwidely applied for the construction of transgenic crops resistant to insect pests. This study aimed to construct novel mutant cry1Ac genes for genetically modified cropswith enhanced insecticidal activities. Using multi-site directed mutagenesis, 34 mutant cry1Ac genes were synthesized and converted at 24 amino acid residues, located on domain I (8 residues) and domain II (16 residues). Thesemutant genes were expressed as a fusion protein with polyhedrin using the baculovirus expression system. The expressed proteins were occluded into polyhedra and activated stably to 65 kDa by trypsin. Among these, Mut-N04, N06 and N16 showed high levels of insecticidal activities against larvae of Plutella xylostella, Spodoptera exigua, and Ostrinia furnacalis. Mut-N16, which showed the highest insecticidal activity, is expected to be a desirable cry gene for introduction into transgenic crops. This study could provide usefulmeans to constructmutant cry geneswith improved insecticidal activities and expand host spectra for transgenic crops.
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