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임현섭,장찬용,배한홍,김준기,이철호,홍진성,주호종,김홍기,Leslie L. Domier 한국식물병리학회 2011 Plant Pathology Journal Vol.27 No.4
Soybean plants infected with Bean pod mottle virus (BPMV) develop acute symptoms that usually decrease in severity over time. In other plant-virus interactions,this type of symptom recovery has been associated with degradation of viral RNAs by RNA silencing, which is accompanied by the accumulation of virus-derived small interfering RNAs (siRNAs). In this study, changes in the accumulation of BPMV siRNAs were investigated in soybean plants infected with BPMV alone, or infected with both BPMV and Soybean mosaic virus (SMV)and in transgenic soybean plants expressing SMV helper component-protease (HC-Pro). In many potyviruses,HC-Pro is a potent suppressor of RNA silencing. In plants infected with BPMV alone, accumulation of siRNAs was positively correlated with symptom severity and accumulation of BPMV genomic RNAs. Plants infected with both BPMV and SMV and BPMV-infected transgenic soybean plants expressing SMV HC-Pro exhibited severe symptoms characteristic of BPMVSMV synergism, and showed enhanced accumulation of BPMV RNAs and siRNAs compared to plants infected with BPMV alone and nontransgenic plants. Likewise,SMV HC-Pro enhanced the accumulation of siRNAs produced from a silenced green fluorescent protein gene in transient expression assays, while the P19 silencing suppressor of Tomato bushy stunt virus did not. Consistent with the modes of action of HC-Pro in other systems, which have shown that HC-Pro suppresses RNA silencing by preventing the unwinding of duplex siRNAs and inhibiting siRNA methylation, these studies showed that SMV HC-Pro interfered with the activities of RNA-induced silencing complexes, but not the activities of Dicer-like enzymes in antiviral defenses.
강호덕,강상구,배한홍,박교수,Kang, Hoduck,Kang, Sang-Gu,Bae, Hanhong,Park, Kyo-Soo,Hall, Richard B. Korean Society of Forest Science 1997 한국산림과학회지 Vol.86 No.3
미류나무의 미성숙 ovule과 줄기로부터 유기된 캘러스에 plasmid pBI221 유전자를 유전자총을 이용하여 인위적으로 삽입하였다. Plasmid pBI221은 CaMV-35S 유전자에 의하여 발현되는 ${\beta}$-glucuronidase(GUS) reporter 유전자를 포함하고 있다. Plasmid pBI221이 물리적으로 삽입된 후 GUS 유전자의 발현정도는 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-glue)의 반응에 의해 분석되었고, 유전자의 일시적 발현현상은 ovule, 캘러스 시료에 X-glue substrate의 반응에 의하여 나타나는 뚜렷한 점(spot)의 수에 따라서 조사하였다. 본 실험에서 particle bombardment후 GUS유전자의 발현검정에 있어 가장 중요한 요인은 bombardment 횟수와 X-glue substrate에 노출된 시간이었다. X-glue substrate와의 반응결과, 캘러스와 ovule에서 각각 56.8%, 75.9%의 반응 점들을 나타냈다. 여러 처리중 두번의 연속적인 shot와 bombardment 이후, X-glue과 sample을 48시간 반응시킨 후 24시간 동안 alcohol로의 침지가 가장 많은 수의 점을 유기시켰고, 이들 반응으로부터 평균 $25.75{\pm}2.77$(ovule), $11.43{\pm}1.22$(calli)개의 반응 점을 보였다. 유전자총에 의한 외래 유전자 도입에 관한 본 연구는 두종류의 시료로부터 빠른 시간 내에 유전자 발현을 볼 수 있을 뿐만 아니라, 지금까지 Agrobacterium을 이용한 형질전환이 보고되지 않은 미류나무(Populus deltoides)의 형질전환 연구에 대체 방법을 제공하리라 생각된다. Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and ${\beta}$-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced about $25.75{\pm}2.77$(per ovule), $11.43{\pm}1.22$(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.
남문,임현섭,배한홍,이봉춘,윤영남,John Hammond,LeslieL.Domier 한국식물병리학회 2013 식물병연구 Vol.19 No.2
To facilitate their spread, plant viruses have developed several methods for dispersal including insect and seed transmission. While insect transmission requires virus stability against insect digestion, seed-transmitted viruses have to overcome barriers to entry into embryos. Bean pod mottle virus (BPMV) is transmitted through seed at levels typically below 0.1%, but co-infection with Soybean mosaic virus (SMV) enhanced the seed transmission rate of BPMV in one experiment. In contrast, the rate of SMV seed transmission was not affected by BPMV co-infection. In a second preliminary study, the rate of SMV transmission was lower in an isoline of Williams 82 that contained a null mutation for the Kunitz trypsin inhibitor gene than in Williams 82. In this preliminary study, we observed that factors such as protease inhibitor expression and dual infection may affect the frequency of seed transmission of BPMV and SMV.
포플러의 미성숙 배와 캘러스에서 유전자총에 의한 GUS - gene 의 일시적 발현
강호덕,강상구,배한홍,박교수,( Ho Duck Kang,Sang Gu Kang,Han Hong Bae,Kyo Soo Park,Richard B . Hall ) 한국산림과학회 1997 한국산림과학회지 Vol.86 No.3
Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and β-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-β-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced abet 25.75±2.77(per ovule), 11.43±1.22(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.
Update on the Effects of Sound Wave on Plants
Md. Emran Khan Chowdhury,배한홍,임현섭 한국식물병리학회 2014 식물병연구 Vol.20 No.1
Plant growth is considered the sum of cell proliferation and subsequent elongation of the cells. The continuousproliferation and elongation of plant cells are vital to the production of new organs, which have a significantimpact on overall plant growth. Accordingly, the relationship between environmental stimuli, such as temperature,light, wind, and sound waves to plant growth is of great interest in studies of plant development. Sound waves can have negative or positive effects on plant growth. In this review paper we have summarizedthe relationship between sound waves and plant growth response. Sound waves with specific frequenciesand intensities can have positive effects on various plant biological indices including seed germination, rootelongation, plant height, callus growth, cell cycling, signaling transduction systems, enzymatic and hormonalactivities, and gene expression.
남지륜,임현섭,남문,배한홍,이철호,John Hammond,이봉춘 한국식물병리학회 2013 Plant Pathology Journal Vol.29 No.4
The Potexvirus Alternanthera mosaic virus (AltMV) hasmultifunctional triple gene block (TGB) proteins, amongwhich our studies have focused on the properties of theTGB1 protein. The TGB1 of AltMV has functionsincluding RNA binding, RNA silencing suppression,and cell-to-cell movement, and is known to form homologousinteractions. The helicase domains of AltMVTGB1 were separately mutated to identify which regionsare involved in homologous TGB1 interactions. Theyeast two hybrid system and Bimolecular FluorescenceComplementation (BiFC) in planta were utilized to examinehomologous interactions of the mutants. Helicasemotif I of AltMV TGB1 was found to be critical tomaintain homologous interactions. Mutations in theremaining helicase motifs did not inhibit TGB1 homologousinteractions. In the absence of homologous interactionof TGB1, subcellular localization of helicasedomain I mutants showed distinctively different patternsfrom that of WT TGB1. These results provide importantinformation to study viral movement and replicationof AltMV.
임현섭,이미연,문제선,문중경,유용만,조인숙,배한홍,Matt deBoer,주호종,John Hammond,Andrew O. Jackson 한국식물병리학회 2013 Plant Pathology Journal Vol.29 No.1
Barley stripe mosaic virus (BSMV) induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed:Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB) treatments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW). BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed:Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3.