Characterization and RT-PCR Detection of dsRNA Mycoviruses from theOyster Mushroom, Pleurotus ostreatus

서장균,임원석,이세원,김국형,유영복,정지해 한국식물병리학회 2004 Plant Pathology Journal Vol.20 No.3

The partial nucleotide sequences of the genomic dsRNAmycovirus infecting Pleurotus ostreatus isolates ASI2223and Suhan were determined and compared with thoseof mycoviruses belonging to partitiviruses and totiviruses. Partial nucleotide sequences of the purified dsRNAfrom ASI2223 and Suhan showed RNA-dependentRNA polymerase sequences that are closely related tothose of partitiviruses, including Fusarium poae virus 1,Fusarium solani virus, Rhizoctonia solani virus, Disculadestructiva virus 2, and Oyster mushroom isometric virus2. Specific primers were designed for RT-PCR detectionof dsRNA viruses from the P. ostreatus isolate ASI2223and Suhan. Two virus specific primer sets were found tospecifically detect each virus among six sets of designedoligonucleotide primers. Collectively, these results suggestthat dsRNA mycoviruses from P. ostreatus isolatesASI2223 and Suhan belong to the family Partitiviridae,although, they are not the same virus species. Ourresults also suggest that these virus-specific primer setscan be employed for the specific detection of each viralsequence in infected tissues.

Infectious in vivo Transcripts from a Full-length Clone of Soybean mosaic virus Strain G5H

서장균,이혁근,최홍수,이수헌,김국형 한국식물병리학회 2009 Plant Pathology Journal Vol.25 No.1

An infectious full-length clone of Soybean mosaic virus (SMV) strain G5H was constructed under the control of the cauliflower mosaic virus 35S promoter. The cloned SMV G5H established infections upon simple rubinoculation of soybean leaves with intact plasmid DNA. We demonstrated that this SMV G5H infectious DNA clone caused typical characteristic symptoms and virulence of SMV strain G5H in twelve tested soybean cultivars. Soybean cultivars Lee74, Somyungkong and Sowonkong developed systemic mosaic symptom while Kwanggyo, Taekwangkong, Hwangkeumkong and Geumjeongkong-1 showed systemic necrosis. In contrast, Geumjeongkong-2, Jinpumkong-2, L29, V94-5152 and Ogden showed resistant response against SMVG5H infection. We also determined full-length sequence of cloned SMV-G5H. The phyogenetic analyses reveal that SMV-G5H is most closely related to SMV-G5, and support that SMV-G5H might be derived from SMVG5 by recombination rather than mutation.

Interaction Study of Soybean mosaic virus Proteins with Soybean Proteins usingthe Yeast-Two Hybrid System

서장균,황성현,강성환,최홍수,이수헌,손승한,김국형 한국식물병리학회 2007 Plant Pathology Journal Vol.23 No.4

Interactions between viral proteins and host proteins are essential for virus replication. Especially, translation of viral genes completely depends on the host machinery. In potyviruses, interactions of genome-linked viral protein (VPg) with host translation factors including eIF4E, eIF(iso)4E, and poly(A)-binding protein (PABP) has previously been characterized. In this study, we investigated interactions between Soybean mosaic virus (SMV) viral proteins and host translation factors by yeast two-hybrid system. SMV VPg interacted with eIF4E, eIF(iso)4E, and PABP in yeast two-hybrid system, while SMV helper component proteinase (HCpro) interacted with neither of those proteins. The interaction between SMV NIb and PABP was also detected. These results are consistent with those reported previously in other potyviruses. Interestingly, we found reproducible and specific interactions between SMV coat protein (CP) and PABP. Deletion analysis showed that the region of CP comprising amino acids 116 to 206 and the region of PABP comprising amino acids 520 to 580 are involved in CP/PABP interactions. Soybean library screening with SMV NIb by yeast twohybrid assay also identified several soybean proteins including chlorophyll a/b binding preprotein, photosystem I-N subunit, ribulose 1,5-biphosphate carboxylase, ST-LS1 protein, translation initiation factor 1, TIRNBS type R protein, RNA binding protein, ubiquitin, and LRR protein kinase. Altogether, these results suggest that potyviral replicase may comprise a multiprotein complex with PABP, CP, and other host factors.

버섯 국산품종 확대보급체계 구축

정종천,이찬중,신평균,서장선 한국버섯학회 2012 버섯 Vol.16 No.2

농촌진흥청 버섯과에서 2009년부터 시작하여 최근에 이르러 성과를 거두고 있는 버섯 국산품종을 전국에 조기 확대보급 함으로써 로열티 부담을 경감하는 시스템 구축 효과에 대해 설명하고자 한다. 우리나라 버섯산업은 2010년도를 기준으로 재배면적 353ha에서 20만톤을 생산하여 총생산액이 8,860억원 수준으로 성장하였으며, 수출실적도 팽이버섯과 새송이를 중심으로 2만2천톤, 5,000만 달러에 이른다. 우리나라의 버섯재배는 70년대의 기업형 양송이 재배, 80년대부터 선진 외국에서도 없는 느타리 균상재배법을 개발하여 농가 부업형태로 추진되어 농가의 중요한 소득원으로 정착하였다. 당시에는 버섯 신품종이 육성되면 전국의 90여개소나 되었던 종균배양소에서 분양받아 전국 버섯재배 농가들에게 빠르게 보급이 되었다. 그러나 최근에는 자체적으로 배양시설을 갖춘 대규모의 병재배시설이 많아짐에 따라 버섯 가격이 대폭 하락하여 균상재배가 크게 줄어들었고 종균배양소도 20여개소만 남아 있는 실정이다. 그나마 남아있는 배양소들간의 경쟁으로 외국에서 도입한 자기 품종만을 농가에 보급하면서 국가에서 육성한 품종들은 점차 사장되어 가는 실정이다. 이러한 상황에서 우리 국립원예특작과 학원에서는 국가에서 개발한 버섯 신품종을 버섯 농가에 조기 보급하고 바로 재배한 결과를 농가와 함께 평가할 수 있는 기회를 제공하고자 본 사업을 추진하게 되었다. 본 사업은 2009년부터 2012년까지 4년간에 걸쳐 느타리 등 19종 34품종의 국산품종을 전국 387개소에 보급함으로써, 인근농가 실증 확산 등 파급효과에 힘입어 국산품종 점유율을 당초 30%에서 2012년도에는 45% 수준으로 끌어 올렸다. 농촌진흥청 및 지자체의 버섯 연구기관에서 새로 육성된 품종은 농촌진흥청의 신품종심의를 거쳐 국립종자원에 품종보호출원을 하고 검증 확인 절차를 밟은 후 일선 버섯 농가에 보급한다. 국립원예특작과학원에서는 버섯 품종목록과 품종의 설명자료를 작성하여 전국의 시군농업기술센터에 배부하고, 시군의 자체시범사업에 연계하여 신청한 품종의 종균을 공급하였다. 또한 자가배양시설을 갖춘 병재배 농가에도 특별히 신청하는 신품종의 접종원을 별도로 분양해 주었다. 본 사업에 대한 반응을 살펴보면 과거에는 버섯재배 농가에서는 지난 몇 년 동안 국가 버섯연구소에서 무슨 신품종이 나왔는지도 모르고, 오로지 거래하는 종균배양소에서 권하는 품종만을 재배해 왔는데 이러한 농촌진흥청의 노력으로 새로운 품종을 직접 재배해 볼수 있는 좋은 기회로 생각하고 이러한 현장의 눈높이에 맞는 시책이 계속 진행 되었으면 좋겠다고들 한다. 일선 시군 농업기술센터의 버섯담당 지도사들은 이러한 시범사업을 추진하면서 지난날에는 단지 농가에서 원하는 품종 위주로 선택 보급하였으나, 본 사업을 통하여 새롭게 제공받은 신품종 특성과 자세한 설명자료, 그리고 연찬회에서의 새로운 인식 등은 각 지역사회의 버섯산업발전에 많은 도움이 되었다고 한다. 또한, 팽이버섯 등 병재배 농가에서는 일본 품종 사용에 대한 대가로 버섯판매액의 5%를 로열티로 요구 받았으나 국산신품종의 접종원을 분양 받아 시험 재배한 이후로는 로열티 요구액도 1% 정도로 크게 낮아졌다고 한다. 버섯 국산 신개발 품종을 일선 농가에 조기 확대 보급하고자 하는 본 사업을 통하여 직접적으로 외국 품종의 재배면적을 줄여서 로열티 지불액을 낮추었으며, 간접적으로 외국품종의 사용에 대한 로열티 협상시 사용료 인하로 버섯 농가의 부담을 낮추는 효과가 있었다. 내년부터는 이러한 버섯 국산품종 조기 보급 시스템의 확대를 더욱 효율적으로 추진하여 국산품종의 점유율을 2011년 40%에서 2017년에는 70% 수준으로 끌어 올릴 수 있도록 하기 위하여 중앙 및 각 도, 시군 지자체의 기술보급사업에 중점 반영하여 확대추진할 것을 건의하였다. 그리고 국립원예특작과학원에서는 매년 국산품종 목록과 특성 설명자료를 작성하여 일선에 배부하고 관계 설명회를 가질 계획이다.

Clinical Significance of Platelet Count at day +60 After Allogeneic Peripheral Blood Stem Cell Transplantation

손상균,백진호,김종광,김동환,서장수,이규보,이난영,원동일 대한의학회 2006 Journal of Korean medical science Vol.21 No.1

Adipose-Derived Regenerative Cell Injection Therapy for Postprostatectomy Incontinence: A Phase I Clinical Study

권태균,최재영,김태환,양정덕,서장수 연세대학교의과대학 2016 Yonsei medical journal Vol.57 No.5

Purpose: We report our initial experience with transurethral injection of autologous adipose-derived regenerative cells (ADRCs) for the treatment of urinary incontinence after radical prostatectomy. Materials and Methods: After providing written informed consent, six men with persistent urinary incontinence after radical prostatectomy were enrolled in the study. Under general anesthesia, about 50 mL of adipose tissue was obtained from the patients by liposuction. ADRCs were obtained by separation with centrifugation using the Celution cell-processing device. A mixture of ADRCs and adipose tissue were transurethrally injected into the submucosal space of the membranous urethra. Functional and anatomical improvement was assessed using a 24-h pad test, validated patient questionnaire, urethral pressure profile, and magneticresonance imaging (MRI) during 12-week follow-up. Results: Urine leakage volume was improved with time in all patients in the 24-h pad test, with the exemption of temporal deteriorationat the first 2 weeks post-injection in 2 patients. Subjective symptoms and quality of life assessed on the basis of questionnaireresults showed similar improvement. The mean maximum urethral closing pressure increased from 44.0 to 63.5 cm H2O at 12 weeks after injection. MRI showed an increase in functional urethral length (from 6.1 to 8.3 mm) between the lower rim of the pubicbone and the bladder neck. Adverse events, such as pelvic pain, inflammation, or de novo urgency, were not observed in any case during follow-up. Conclusion: This study demonstrated that transurethral injection of autologous ADRCs can be a safe and effective treatment modalityfor postprostatectomy incontinence.

다발성 신품종 느타리 ‘다굴’의 육성 및 자실체 특성

신평균,김희정,최찬식,유영복,공원식,장갑열,오연이,정종천,서장선,오세종,이금희 한국버섯학회 2013 한국버섯학회지 Vol.11 No.3

To develop a new variety of oyster mushroom(Pleurotus ostreatus), parental strains was selected by the method of Mon-Mon crossing between monokaryotic strains derived from ASI 2596(Suhan No.3) and ASI 2782(Black pileus mutant). The SB-73(ASI 2596-11 x 2782-8) was shown the best cultural characteristics, selected to be a new variety and named as ‘Dagul’. The ‘Dagul’ was formed incompatibility line distinctly in the confrontation growth of parental strains Suhan No.3 and ASI 2782. The optimum temperature for mycelial growth, fruiting body development and pH arrange were 25~30oC, 14~17oC and pH5~8, respectively. Fruiting body production per bottle was about 68.024.1 g which is almost 115% quantity compared to that of other variety Suhan No.3. And also the stipe is long and individual generation is multiple. Analysis of the genetic characteristics of the new variety ‘Dagul’ showed different DNA bands as that of the control strains, Suhan No.3 and ASI 2782, when RAPD(Random Amplified Polymorphic DNA) primers URP7 and Rcb1 were used. This new variety 'Dagul' of oyster mushroom is characterized by multiple of individual generation and the stipe is long. We therefore expect that this new strain will increase of the income by cultivation of field.

Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered of Zucchini yellow mosaic virus

강민지,서장균,최호성,최홍수,김국형 한국식물병리학회 2016 Plant Pathology Journal Vol.32 No.1

The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMVbased viral vector in association with the simple rubinoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits

Establishment of an Agrobacterium-mediated Inoculation System for Cucumber Green Mottle Mosaic Virus

강민지,서장균,송다미,최홍수,김국형 한국식물병리학회 2015 Plant Pathology Journal Vol.31 No.4

The infectious full-length cDNA clones of Cucumber green mottle mosaic virus (CGMMV) isolates KW and KOM, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35S promoter. We successfully inoculated Nicotiana benthamiana with the cloned CGMMV isolates KW and KOM by Agrobacterium-mediated infiltration. Virulence and symptomatic characteristics of the cloned CGMMV isolates KW and KOM were tested on several indicator plants. No obvious differences between two cloned isolates in disease development were observed on the tested indicator plants. We also determined full genome sequences of the cloned CGMMV isolates KW and KOM. Sequence comparison revealed that only four amino acids (at positions 228, 699, 1212, and 1238 of the replicase protein region) differ between the cloned isolates KW and KOM. A previous study reported that the isolate KOM could not infect Chenopodium amaranticolor, but the cloned KOM induced chlorotic spots on the inoculated leaves. When compared with the previously reported sequence of the original KOM isolate, the cloned KOM contained one amino acid mutation (Ala to Thr) at position 228 of the replicase protein, suggesting that this mutation might be responsible for induction of chlorotic spots on the inoculated leaves of C. amaranticolor.

동종 말초혈액 조혈모세포를 이용한 다양한 임상적용에 관한 단일병원 경험보고

채의수,손상균,성우진,서광운,박성원,김종광,정진태,이난영,서장수,이규보 경북대학교 병원 2002 경북대학교병원의학연구소논문집 Vol.6 No.1

배경:동종 말초혈액 조혈모세포 이식은 골수이식에 비해 많은 양의 조혈모세포 및 T―림프구 이식으로 인해 빠른 회복이 가능하며 강력한 이식편대종양반응이 유도될 수 있는 장점이 있어 이를 잘 응용한다면 악성 혈액질환에서 치료율을 높일 수 있을 것으로 사료된다.본 연구에서는 동종 말초혈액 조혈모세포 이식의 장점을 이용한 다양한 임상적용을 소개하고자 한다. 방법:경북대학교병원 조혈모세포 이식센터에서 동종 말초혈액 조혈모세포 이식을 시행한 증례 중에서 동종 골수이식수술에 비해 말초혈액 조혈모세포 이식이 유리할 것으로 생각디는 재발의 가능서이 높은 악성질환에서의 더욱 강력한 이식편대종양반응 유도를 위한 동종 말초혈액 조혈모세포를 이용한 예방적 공여자 림프구주입 등의 임상적용을 시도한 증례 및 결과를 분석하였다. 결과:17례의 재발의 가능성이 높은 고위험군의 악성 혈액질환을 대상으로 한 동종 말초혈액 조혈모세포 이식에서 90일 이내의 치료와 관련된 사망률은 23.5%이었고 중앙 추적기간 563일(범위 120∼926일)간의 무병생존율은 35.3%이었다.CSF로 가동화한 말초혈액 조혈모세포를 추가적으로 수혈받은 환자 7명 중 4명(57.1%)이 중앙 추적기간 538일(범위120∼926일)째에 완전관해 상태로 생존 중에 있다.그 외 말초혈액 조혈모세포를 아용한 미니 이식,동종 말초혈액 조혈모세포를 이용한 예방적 stepwise dose-increment 공여자 림프구주입술 등을 시행한 증례에서 장기 생존의 결과를 보여주고 있다. 결론:동종 말초혈액 조혈모세포 이식은 많은 양의 조혈모세포 및 T-림프구의함유로 인해 높은 생착률과 빠른 회복,강력한 이식편대종양반응 등이 유도되는 장점이 있으므로 기존의 골수이식술에 비해 보다 광범위한 임상적용이 가능하리라 사료된다. Background: It is apparent that more stem cells can be harvested by mobilization treatment with recombinant human G-CSF and/or GM-CSF from normal healthy donors in allogeneic peripheral biood stem celltransplantation(allo-PBSCT) compared to allogeneic bone marrow transplantation(allo-BMT).It is also known to be more effective in inducing the graft-vs-tumor affects than allogeneic BMT because of higher T cell content. Methods: A variety of clinical applications with allo-PBSCT was done for patients with hematoligical malignancies with a high risk of relapse in sigle transplantation center. We reported the preliminary results on trial of allo-PBSCT followed by planned prophyactic G-and/or GM-CSF primed donor lymphyocyte infusion additionally reserved at harvest in hematological malignancies with a high risk of relapse and also presented the successful trial of non-myeloablative transplantation for old aged AML paient in 4th complete remission and cases with 2nd transplantation with allo-PBSCs. Results: Seventeen patients with hematological malignancies with a high risk of relapse were enrolled in trial of allo-PBSCT followed by prophylactic donor lymphocyte infysion.All patients received allogeneic PBSCT from HLA- matched sibling donor.Aeven out of 17 patients recived additional PBSCs with a median number of CD3+ CELLS OF 5.0×10^7/kg(range,3.0 to 9.9×10^7/kg), between day 41 and day 120.Four surving patients(4/7) were free of disease when last assessed(median follow-up duration, 538 days), but were suffering from chronic GVHD(1 limited and 3 extensive). A 56 year old acute myeloid leukemia patient in the 4th complete remission was successfully treated with allo-PBSCT with non-myeloablative conditioning regimen.One of 2 patients who received second transplantation with allo-PBSCT has shown a long term disease free survival. Conclusion: A merit of allo-PBSCT would allow us to design a variety of clinical applications. Allo-PBSCT might be preferable in special clinical setting such as non-myeloablative transplantation, second transplantation,or the situation in need of the strong GVL effect. And also CSF-primed PBSCs can be used for the purpose of donor lymphocyte infusion.(korean J Hematol 2001;36:214~222)