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Sihyeon Kim,Se Jin Lee,Yi-Ting Yang,Jae Su Kim 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.04
Bean bug, Riptortus pedestris (Hemiptera: Alydidae) is an agriculturally serious pest in East Asian countries. Chemical pesticides have been contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Herein entomopathogenic Beauveria bassiana JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by contact-exposure and spray methods. The Agrobacterium tumefaciens-mediated transformation of B. bassiana JEF-007 generated random transformants and some mutants showed reduced virulence against Tenebrio molitor (Coleoptera: Tenebrionidae) larvae and R. pedestris nymph. Compared to the wild-type, the two transformants showed remarkably different morphology, conidial production, and thermotolerance. To figure out pathogenicity-related genes, thermal asymmetric interlaced (TAIL) PCR of the random transformants was performed and possibly some virulence-related genes were predicted. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.
Micro-groove patterned PCL patches with DOPA for rat Achilles tendon regeneration
Sihyeon Park,이민석,Jin Jeon,Juhan Lim,조현철,Suk Ho Bhang,Hee Seok Yang 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.105 No.-
Achilles tendon regeneration using current tissue engineering techniques requires morphometricallymimetic tendon biomaterials to attain the required microstructural tendon tissue and to achieve the necessarymechanical properties. In this study, we used nano- (400 nm) and micro-groove (5000 nm) patternedpolycaprolactone patches (PCL-400, PCL-5000), which offer adequate elastic modulus andbiocompatibility. The PCL patches were fabricated via capillary force lithography and were surface modifiedwith 3,4-dihydroxy-L-phenylalanine (DOPA) for increased hydrophilicity. We hypothesized that theresulting biologically surface-modified elastic groove patterned patches would enhance the cell behaviorsin vitro and the rat Achilles tendon regeneration in vivo. The micro-groove patterned PCL patcheswith DOPA coating (DOPA-PCL-5000) exhibited excellent cell elongation, tenogenic differentiation andYAP expression of the mesenchymal stem cells in our in vitro study and these results were further confirmedby application to the rat Achilles tendon rupture model in our in vivo study. DOPA-PCL-5000induced dramatic regeneration of rat Achilles tendon compared to the other groups, which was furtherconfirmed by the Achilles functional index and histological analysis at 8 weeks. In conclusion, DOPAPCL-5000 can be used not only for damaged Achilles tendon, but also for various functional tendon regenerationtechniques such as rotator cuff, patellar tendon, and flexor tendon.
Establishment of entomopathogenic fungal virulence assay against Protaetia brevitarsis seulensis
Sihyeon Kim,Seok Ju Lee,So Eun Park,Jong Cheol Kim,Mi Rong Lee,Se Jin Lee,Jae Su Kim 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.04
Many turfgrass pests, Ectinohoplia rufipes, Exomala orientalis and Popillia quadriguttata, cause serious damage to grass fields in Korea. In this study, control of turfgrass pests using entomopathogenic fungi were investigated using Protaetia brevitarsis seulensis as an alternative insect. In a preliminary experiment, some Metathizium sp. showed ca. 60% virulence after 7 days exposure to the entomopathogenic fungi. Compared to the contact-exposure, overall a spray treatment showed lower control efficacy against P. brevitarsis seulensis. Furthermore, mortality of various developmental stages of P. brevitarsis seulensis was investigated under laboratory conditions. This work suggests that when treated with fungi mortality of P. brevitarsis seulensis was closely related with developmental stage. These results provide a basic information for the control of P. brevitarsis seulensis using entomopathogenic fungi
Sihyeon Kim,Se Jin Lee,Jeong Seon Yu,Sung Duk Seo,Han Wool Cho,Yu-Shin Nai,Jae Su Kim 한국응용곤충학회 2014 한국응용곤충학회 학술대회논문집 Vol.2014 No.04
Bean bug, Riptortus clavatus Thunberg (Heteroptera: Alydidae) causes serious damage to Leguminosae. Herein an entomopathogenic fungal virulence assay system against bean bugs was established to construct a fungal database which can be used in integrated pest management (IPM). First to obtain as many bean bugs as possible at the same stage, host plant-preference and developmental synchronization of bean bugs were investigated. In the preference assay, five pairs of adults were infested in a plastic cage, where a pot of green bean, pea or cowpea was previously placed. The highest fecundity and the fastest development of bean bug was observed in the green bean cage. Secondly, in the synchronization experiment, eggs were collected from the cage of adults in 1, 3, 5 and 7 days after oviposition and transferred to a fresh cage with green beans. From the every 4 days of survey, similar stages of bean bugs were found in the cages with the oviposition for 1 and 3 days, rather than the longer times of oviposition. A fungal bioassay against bean bugs was conducted using the bean bugs from the above insect rearing system. Ten Beauveria bassiana isolates were cultured on quarter-strength Sabouraud dextrose agar (¼SDA) for 7 days at 25°C. Ten 4th instar of nymphs were placed on a cultured plate for 1 hour and tranferred to a fresh moisturized plate with grains of green bean. ERL836 isolate treatment showed the highest virulence and fungal mycosis was observed on the bean bugs. In conclusion, these results can be useful to establish an entomopathogenic fungal database for IPM.
Virulence factors of bean bug (Riptortus pedestris)-pathogenic Beauveria bassiana JEF-007
Sihyeon Kim,Se Jin Lee,Mi Rong Lee,Min Ho Song,Yi-Ting Yang,Jae Su Kim 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10
Bean bug, Riptortus pedestris is an agriculturally serious pest in East Asian countries, reducing the value of crop quality and loss of income in agribusiness. Chemical pesticides have contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Beauveria bassiana and Metarhizium anisopliae JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by spray and contact-exposure methods. Supernatant showed different level of enzyme activity including chitinase, Pr1 protease and lipase. The Agrobacterium tumefaciens-mediated transformation generated random transformants and some mutants had reduced virulence. TAIL-PCR of the random transformants revealed virulence-related genes. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.
hpRNA-induced gene silencing in Beauveria bassiana JEF-007 for gene characterization
Sihyeon Kim,Se Jin Lee,Mi Rong Lee,Jong Cheol Kim,So Eun Park,Jae Su Kim 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.10
A genus of entomopathogenic Beaveria bassiana has been widely used in pest management, however little studies havebeen given to its virulence-related genes. To identify the roles of virulence genes, AtMT-induced random mutants weregenerated and followed by localization study with TAIL-PCR. Two genes, Complex I intermediated-associated protein30 (CIP30) and Autophagy protein 22 (Atg22), were predicted as virulence-related genes in B.bassiana JEF-007. To validatethe a possible relationship between two genes and fungal virulence, hpRNAi was performed. A hpRNAi plasmid wasconstructed as a model system to knock down of egfp gene in egfp-expressing B. bassiana transformant. A Real-TimePCR showed the knock down of egfp gene expression via hpRNAi. The CIP30 and Atg22 genes in B. bassiana willbe validated by the established RNAi technique. This work can be a strong platform for the functional genetics in B.bassiana.
Sihyeon Kim,Se Jin Lee,Mi Rong Lee,Min Ho Song,Jae Su Kim 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.04
Bean bug, Riptortus pedestris is an agriculturally serious pest in East Asian countries, reducing the value of crop quality and loss of income in agribusiness. Chemical pesticides have contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Beauveria bassiana and Metarhizium anisopliae JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by spray and contact-exposure methods. The isolates produced high levels of pathogenesis-related enzymes, such as chitinase, Pr1 protease and lipase. The Agrobacterium tumefaciens-mediated transformation generated random transformants and some mutants had reduced virulence against bean bugs, which provided some materials to figure out pathogenicity-related genes in the fungi. Now characterization of flanking region of the integrated fragment is underway and this work may reveal some important genes in the pathogenesis. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.
Sihyeon Kim,Jong Cheol Kim,Mi Rong Lee,So Eun Park,Dongwei Li,Sehyeon Baek,Laila Gasmi,Min Sung Jo,Tae Young Shin,Junheon Kim,Jae Su Kim 한국응용곤충학회 2019 한국응용곤충학회 학술대회논문집 Vol.2019 No.04
Insect killing fungus Beauveria bassiana has been widely studied as a biological control agent. However, many studies have been focused on lab or field-based management. Herein this work, comparison of three B. bassiana strains was investigated under a molecular level. The whole genome sequences of ERL836, JEF-007 were analyzed by PacBio (35.5 Mb of ERL836 and 36.5 Mb of JEF-007) and ARSEF2860 referenced from GenBank (33.7 Mb). To compare the three strains, virulence, thermotolerance and chemical resistance were assayed. The transcriptomes of non-infecting B. bassiana and infecting B. bassiana against western flower thrips were analyzed using RNA-seq. This work can provide that genome features, functions, morphology and gene expression could be different under the molecular level, even if in the same species.