Symposia / S3-5 : Modulation of actin organization during fruit ripening : Expression of a grape cDNA encoding an actin - depolymerizing factor during berry development

피재호(Jae Ho Pyee),(Ho Rim Lee),(Yoo Li Kim) 한국생화학분자생물학회 (구 한국생화학회) 1999 생화학분자생물학회 춘계학술발표논문집 Vol.- No.-

포도 VIASR 유전자 프로모터의 분리 및 발현 분석

길준영,피재호,Kihl, Joon-Yeong,Pyee, Jae-Ho 한국생명과학회 2007 생명과학회지 Vol.17 No.8

VvMSA, a grapevine ASR which is highly inducible by sugar and abscisic acid signals was previously shown to be a transcription factor for a hexose transporter gene VvHT1. We isolated a cDNA clone, VlASR which is regulated temporally during the grape berry development by ACP RT-PCR (annealing control primer reverse transcriptase-polymerase chain reaction) and it proved identical to VvMSA. RT-PCR and real-time PCR analyses revealed that the VlASR gene was expressed in berries at fruit set and that its expression increased as berries aged but decreased at the late ripening stage. In order to understand the regulatory mechanism of the asr gene, a genomic fragment was cloned from grapevine. The genomic DNA was 1375 bp long and a sugar box (sucrose box 3 and sucrose responsive element 1) was identified in the 611 bp upstream region of the open reading frame. Analysis of the VlASR promoter::reporter gene fusion demonstrated that this promoter was expressed in transgenic Arabidopsis even without sucrose treatment. This result suggests that the ASR/VvHT1-mediated sugar/ABA signaling, previously reported in grapevine, may not function in Arabidopsis which has no ASR homologue. 포도 ASR (VvMSA) 단백질은 hexose transporter 유전자 VvHT1의 전사를 조절하는 조절 인자 중의 하나로서 sugar 및 abscisic acid (ABA) 신호에 의해 발현이 유도된다. 본 연구진은 ACP RT-PCR (annealing control primer reverse transcriptase-polymerase chain reaction) 방법을 이용하여 포도 과실발달 과정에서 조절되는 유전자 중 VvMSA와 동일한 cDNA (VlASR)를 클로닝하였다. 이 유전자는 착과 시기에 발현되기 시작하여 과실이 발달하면서 점점 증가하여 착과 후 10 주에 가장 많이 발현되며, 숙기 후반에는 도리어 발현양이 감소하였다. 포도 asr 유전자의 조절기작을 밝히기 위해, 이 유전자의 genomic clone을 분리하였다. 총 1375 bp로 이루어진 이 유전자 절편에는 open reading frame과 100 bp의 intron을 포함하고 있다. 약 600 bp 길이의 프로모터 내에는 sugar 신호전달과 연관이 있는 것으로 알려진 sugar box(sucrose box 3 +sucrose response box 1)가 있다. 프로모터 절편을 reporter 유전자와 연결하여 Arabidopsis에 도입하고 형질전환체를 분석한 결과, reporter 유전자는 sucrose 처리와 상관없이 항상 발현되었다. 이러한 결과는 포도에서 보고된 ASR/VvHT1를 매개로 하는 sugar/ABA 신호전달계가 asr 유전자가 없는 Arabidopsis에서는 작동되지 않음을 시사하고 있다.

Genetic and Physiological Discrepancies from Isolates of Sclerotinia homoeocarpa causing Zoysiagrass Dollar Spot Disease

박대섭,김경덕,길준영,피재호,Park, Dae-Sup,Kim, Kyung-Duck,Kihl, Joon-Yeong,Pyee, Jae-Ho Turfgrass Society of Korea 2006 한국잔디학회지 Vol.20 No.1

난지형 잔디인 한국 안양잔디에서 달라스팟의 병원균인 Sclerotinia honoeocarpa의 isolate, Scz1이 최근 새롭게 동정되었다. Scz1은 한지형 잔디인 크리핑 벤트그래스에서 분리된 표준 균주인 Scb1과는 다른 균사의 색상, 균사간의 친밀도 그리고 병 기주 특이성을 가지는 것으로 알려졌다. 본 연구에서는, Scz1, Scz2(난지형 잔디에서 분리한 또 다른 달라스팟 병원균) 그리고 Scb1을 분자생물학적인 연구, internal transcribed spacer(ITS) 와 random amplified polymorphic DNA(RAPD) assays를 이용하여 동정 및 유전자적 차이를 알아보았다. ITS 실험의 결과, 3개의 isolates가 ITS 부분적 염기 서열 비교 BLAST에 등록되어 있는 S. homoeocarpa의 ITS 염기 서열과 $94{\sim}97%$의 동일성을 지니는 것으로 밝혀졌다. RAPD 실험 결과로는, Scz1과 Scb1의 similarity matrix 범위는 0.167이였고, Scz2와 Scb1은 0.139 그리고, Scz1과 Scz2은 0.713이였다. 계통수(系統樹) 결과는 Scb1과는 달리 Scz1과 Scz2는 유전적으로 높은 동일성을 지니고 있어, 같은 분류에 속한다는 것을 알 수 있었다. 달라스팟 병원균 억제에 효과적인 농약인 프로피코나졸에 대한 $EC_{50}$은 Scz1은 0.012 ${\mu}g/ml$, Scz2은 0.003 ${\mu}g/ml$ 그리고 Scb1은 0.030 ${\mu}g/ml$이었다. 상기 결과로, 동일 병원 기주성과 유사한 유전적 친밀성을 보인 Scz1과 Scz2는 S. homoeocarpa의 동일 그룹에 속하였으나 농약 민감도에서는 차이점을 보였다는 것을 알 수 있었다. 향후, 보다 더 많은 한지형과 난지형 잔디에서 분리된 병원균들을 이용하여 유전적 다양성을 밝히는 연구가 진행되어야 할 것이다. Scz1, an isolate of Sclerotinia homoeocarpa, was recently reported as a novel pathogen responsible for dollar spot disease in Zoysiagrass, a warm season turfgrass. Scz1 possessed different characteristics on mycelial pigment, mycelial affinity and host pathogenecity compared to those of Scb1, a typical isolate, obtained from creeping bentgrass, a cool season turfgrass. In this study, only three isolates, Scz1, Scz2(another analogous isolate of Sclerotinia homoeocarpa from zoysiagrass), and Scb1, were examined at the molecular level using the internal transcribed spacer(ITS) and random amplified polymorphic DNA(RAPD) assays to verify their identification and genetic variation. As a result of ITS assay, partial ITS sequences of three isolates showed 94-97% similarity with a standardized ITS sequence of S. homoeocarpa registered on BLAST. In the analysis of RAPD, range value through similarity matrix was 0.167 between Scz1 and Scb1, 0.139 between Scz2 and Scb1, and 0.713 between Scz1 and Scz2, respectively. Furthermore, tendegram analysis indicated that Scz1 and Scz2, unlike Scb1, were clustered together as accompanying a high genetic similarity. In in vitro fungicide bioassay, $EC_{50}$ value representing the sensitivity degree to propiconazole, a well-known fungicide for dollar spot disease, was 0.012 ${\mu}g/ml$ for Sczl, 0.003 ${\mu}g/ml$ for Scz2, and 0.030 ${\mu}g/ml$ for Scb1. From all data taken, we concluded that both Scz1 and Scz2 belonged to one group of S. homoeocarpa, since they exhibit the same host range and high level of genetic similarity, whereas their chemical competences to a fungicide were different. This study would provide further approach for assessing genetic diversity of S. homoeocarpa isolates as well as characterizing individual isolate against chemical exposure.

유기 패시베이션 박막이 P3HT:PC₆₁BM 활성층을 갖는 고분자 태양전지의 특성에 미치는 영향

이상희,박병민,조양근,장호정,정재진,피재호,Lee, Sang Hee,Park, Byung Min,Cho, Yang Keun,Chang, Ho Jung,Jung, Jae Jin,Pyee, Jaeho 한국마이크로전자및패키징학회 2014 마이크로전자 및 패키징학회지 Vol.21 No.4

재생에너지 광소자로서 스마트 농장 등의 에너지원으로서 고분자 태양전지의 응용이 기대되며 향후 상업화를 위해 효율과 신뢰성 개선이 요구된다. 본 연구에서는 유기 패시베이션 박막을 가지는 헤테로정션 고분자태 전지를 제작하고, 패시베이션 박막이 고분자 태양전지의 특성에 미치는 영향을 조사하였다. 사용된 패시베이션 유기재료로는 폴리비닐알코올과 이크롬산 암모늄을 혼합하여 용해한 후 스핀코팅방법으로 P3HT:$PC_{61}BM$/LiF/Al 기판위에 코팅하여 소자를 제작하였다. 제작된 소자구조는 glass/ITO/PEDOT:PSS/P3HT:$PC_{61}BM$/LiF/Al/passivation layer 이며, 140시간 공기 중에 노출 후 전기적 특성을 측정, 비교한 결과, 패시베이션 처리된 고분자 태양전지가 패시베이션 박막 처리되지 않은 소자에 비해 보다 우수한 전기적 특성을 보여주었다. 즉, 패시베이션 처리된 소자의 전력변환효율은 제작직후 3.0%에서 140시간 노출 후 1.3%로 감소한 반면 패시베이션 처리되지 않은 소자의 경우는 동일한 노출조건에서 3.5%에서 0.1%로 급격한 특성저하를 나타내었다. It is required to improve the efficiency and the reliability of the polymer solar cells (PSCs) as the energy saving optical device for the future application of the smart farm facilities. In this study, we fabricated the bulk hetero junction PSCs with organic passivation film layer for the reliability improvement of the devices. The effects of the passivation layer on the electrical properties of the PSCs were studied. The materials of passivation layer are composed of poly vinyl alcohol (PVA) and ammonium dichromate, and the passivation films were fabricated by the spin coating method on the P3HT:$PC_{61}BM$/LiF/Al substrate. The prepared structure of the device is the glass/ITO/PEDOT:PSS/P3HT:$PC_{61}BM$/LiF/Al/passivation layer. The performances of the PSCs with the organic passivation film showed better electrical properties compared with the PSCs without passivation layers. The power conversion efficiency (PCE) values of passivated PSCs decreased from 3.0 to 1.3% after air exposure for 140 hrs. In contrast, the PCE values for the devices without passivation decreased sharply from 3.5 to 0.1% under the same exposure condition.

Genetic and Physiological Discrepancies from Isolates of Sclerotinia homoeocarpa causing Zoysiagrass Dollar Spot Disease

Dae-Sup Park(박대섭),Kyung-Duck Kim(김경덕),Joon-yeong Kihl(길준영),Jae-Ho Pyee(피재호) 한국잡초학회·한국잔디학회 2006 Weed & Turfgrass Science Vol.20 No.1

난지형 잔디인 한국 안양잔디에서 달라스팟의 병원균인 Sclerotinia homoeocarpa의 isolate, Scz1이 최근 새롭게 동정되었다. Scz1은 한지형 잔디인 크리핑 벤트그래스에서 분리된 표준 균주인 Scb1과는 다른 균사의 색상, 균사간의 친밀도 그리고 병 기주 특이성을 가지는 것으로 알려졌다. 본 연구에서는, Scz1, Scz2(난지형 잔디에서 분리한 또 다른 달라스팟 병원균) 그리고 Scb1을 분자생물학적인 연구, internal transcribed spacer(ITS) 와 random amplified polymorphic DNA(RAPD) assays를 이용하여 동정 및 유전자적 차이를 알아보았다. ITS 실험의 결과, 3개의 isolates가 ITS 부분적 염기 서열 비교 BLAST에 등록되어 있는 S. homoeocarpa 의ITS 염기 서열과 94~ 97%의 동일성을 지니는 것으로 밝혀졌다. RAPD 실험 결과로는, Scz1과 Scb1의 similarity matrix 범위는 0.167이였고, Scz2와 Scb1은 0.139 그리고, Scz1과 Scz2은 0.713이였다. 계통수(系統樹) 결과는 Scb1과는 달리 Scz1과 Scz2는 유전적으로 높은 동일성을 지니고 있어, 같은 분류에 속한다는 것을 알 수 있었다. 달라스팟 병원균억제에 효과적인 농약인 프로피코나졸에 대한 EC??은 Scz1은 0.012 ㎍/㎖, Scz2은 0.003 ㎍/㎖ 그리고 Scb1은 0.030 ㎍/㎖이었다. 상기 결과로, 동일 병원 기주성과 유사한 유전적 친밀성을 보인 Scz1과 Scz2는 S. homoeocarpa의 동일 그룹에 속하였으나 농약 민감도에서는 차이점을 보였다는 것을 알 수 있었다. 향후, 보다 더 많은 한지형과 난지형 잔디에서 분리된 병원균들을 이용하여 유전적 다양성을 밝히는 연구가 진행되어야 할 것이다. Scz1, an isolate of Sclerotinia homoeocarpa, was recently reported as a novel pathogen responsible for dollar spot disease in Zoysiagrass, a warm season turfgrass. Scz1 possessed different characteristics on mycelial pigment, mycelial affinity and host pathogenecity compared to those of Scb1, a typical isolate, obtained from creeping bentgrass, a cool season turfgrass. In this study, only three isolates, Scz1, Scz2(another analogous isolate of Sclerotinia homoeocarpa from zoysiagrass), and Scb1, were examined at the molecular level using the internal transcribed spacer(ITS) and random amplified polymorphic DNA(RAPD) assays to verify their identification and genetic variation. As a result of ITS assay, partial ITS sequences of three isolates showed 94-97% similarity with a standardized ITS sequence of S. homoeocarpa registered on BLAST. In the analysis of RAPD, range value through similarity matrix was 0.167 between Scz1 and Scb1, 0.139 between Scz2 and Scb1, and 0.713 between Scz1 and Scz2, respectively. Furthermore, tendegram analysis indicated that Scz1 and Scz2, unlike Scb1, were clustered together as accompanying a high genetic similarity. In in vitro fungicide bioassay, EC?? value representing the sensitivity degree to propiconazole, a well-known fungicide for dollar spot disease, was 0.012 ㎍/㎖ for Scz1, 0.003 ㎍/㎖ for Scz2, and 0.030 ㎍/㎖ for Scb1. From all data taken, we concluded that both Scz1 and Scz2 belonged to one group of S. homoeocarpa, since they exhibit the same host range and high level of genetic similarity, whereas their chemical competences to a fungicide were different. This study would provide further approach for assessing genetic diversity of S. homoeocarpa isolates as well as characterizing individual isolate against chemical exposure.

포도과피에 특이적으로 발현하는 stilbene synthase cDNA의 분리 및 과실발달 단계별 발현양상 분석

피재호,이호림 단국대학교 1999 論文集 Vol.34 No.-

Resveratrol (stilbene), a phytoalexin, is produced by stilbene synthase from malonyl-CoA and p-coumaroyl-CoA as substrates and its biosynthesis is induced by environmental stresses such as fungal elicitor and UV. In order to elucidate its role as an anti-stress compound, stilbene synthase (stsy) cDNAs were isolated from Jyoho grape skin, which has been known to contain high quantity of resveratrol, and their expressions were analyzed during the berry development. RT-PCR analysis showed that stilbene synthase gene was expressed in higher level in leaves and flowers than in berries, and its expression in berries was specific to skin. The expression was first detected 10 weeks postflowering in berry skin and increased as berries grew and ripened. Interestingly, another homolog was amplified from the RT-PCR analysis. Its expression was observed at the early developmental stage and reached at the maximum level 8 weeks postflowering, but was not detected thereafter. Both of the PCR produts were subcloned and they shared 70% homology in the nucleotide sequence. These results suggest that there are two or more stilbene synthase genes differentially regulated during the berry develepment.

Streptomyces coelicolor A3(2)의 actinorhodin생성에 미치는 아미노산의 영향

김재헌,피재호,오충훈,최정은 단국대학교 1996 論文集 Vol.30 No.-

We investigated the fatty acid compositions before and after the actinorhodin producrion in Streptomyces coelicolor A3(2). The relatice proportion of 15 and 17 carbon branched-chain fatty acids decreased signifi-cantly during the actinorhodin production. Fatty acid compositions in the mycelia grown in the presence of L-arginine as a sole nitrogen source, which supported actinorhodin production, showed higher relative pro-portion of the i15:0, i17:0 and ai17:0 branched-chain fatty acids. Therefore, it was concluded that common precursors derived from the amino acid were used for actinorhodin and fatty acid biosynthesis.

뿌리혹 특이 단백질 Nodule - 24 의 담배에서의 발현

이석찬,피재호,윤은경,구영연,천종윤,천충일 한국유전학회 2000 Genes & Genomics Vol.22 No.3

Nodulin-24 is a soybean nodulin that is expressed specifically in root nodules. It was shown to have a cleavable signal sequence and localized to a symbiosome membrane. For understanding the targeting process of nodulin-24 to the symbiosome, two different sizes of nodulin-24 cDNAs were fused to β-glucuronidase (GUS) gene and the fused constructs were introduced into tobacco plant. The presence of the fusions in transgenic plants was confirmed by polymerase chain reaction (PCR). Histochemical staining of mature tobacco leaves with X-glucuronide did not show the expression of the nodulin-24/GUS fusions. However, their expression was found in apical meristems and roots of the transgenic tobacco plants. Northern analysis of mature leaves showed the presence of transcripts for the fusions. These results indicate that the fusions were expressed, and the products may be targeted to vacuole and degraded in mature leaves, but that they were not degraded in the maturing tissues.