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벼 (Oryza sativa L.)배양세포의 고중력유도성 cDNA의 탐색
권순태,김길웅,Kiyoharu OONO 한국식물생명공학회 1994 식물생명공학회지 Vol.21 No.2
벼(Oryza sativa L. cv Nipponbaie)배양세포에 중력 450,000 x g를 처리하여 cDNA library를 만들고, 무처리 및 고중력을 처리한 cDNA 프로브로 스크리닝을 실시하여 고중력에 특이적으로 양성반응을 나타내는 GSC 13 및 GSC124 cDNA를 선발하였다. 선발된 두 유전자 GSC 13 및 GSC 124의 길이는 각각 1.34 및 0.67 kilobase pairs였으며, 배양세포내에서 관련된 transcript의 크기는 각각 2.0 및 1.9 kilobase pairs인 것으로 나타났다. 두 유전자를 프로브로한 Northern hybridization을 실시한 결과 GSC 13, GSC 124 공히 배양세포내에 고중력처리에 의해 특이적으로 축적되는 mRNA가 나타났으며, 중력강도 300,000 x g 에 비해 450,000 x g 처리에서 더욱 강한 축적을 보였고 450,000 x g 4시간 처리에서 최대의 수준을 보였다. Two different gravity specific cDNA, namely, GSC 13 and GSC 124 with length of 1.34 and 0.67 kilobase pairs, and transcripts of 2.0 and 1.9 kilobase pairs, respectively. were isolated by differential screening and northern hybridization of the total RNA isolated from treated and untreated cultured cells showed that maximum levels of trannscripts were achieved after 4 h of gravity stress at 450, 000 x g for both, GSC 13 and GSC 124, suggesting that these mRNA could be expressed and translated into polyeptites related to the cell to extream gravity stress.
식물배양세포의 저장관계 유전자탐색과 형질전환에의 활용 1. 벼 배양세포의 건조보존기구와 관련유전자 선발
김길웅,대야청춘,신동현,Virigool, S.,Shinzaki, K. Y.,홍석영 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
A system for long-term dry preservation of rice (Oryza sativa L.) callus and a protection mechanism regulating the survival of dried callus were investigated. The highest survival of dried callus and the highest regeneration of plantlets were observed in calli which had been pretreated with 10^(-5) M abscisic acid (ABA) in the presence of 90 g/L of sucrose. A corresponding accumulation of the RNA of the rab 16A gene (a rice gene induced by ABA and water stress) was detected in dried callus, mature seeds, and callus pretreated with 10^(-5) M ABA. Analysis of protein by two dimensional polyacrylamide gel electrophoresis(2D-PAGE) demonstrated different protein patterns in dried callus pretreated with 10-s M ABA and 90 g/L of sucrose compared to callus dried without the pretreatment. Three different cDNA clones, pDHS1, pDHS2, and pDHS3 contained cDNA inserts in size of 3.8, 3.2, and 3.2 kilobase pairs, respectively, were isolated by using Nhe I fragment from rab 16A as a probe. All the three clones exhibited unique restriction maps.
벼 RAPD 표지 이용 低溫幼苗生育관여 量的形質遺傳子座(QTL) 分析
Kyung Min Kim(金敬旻),Jae Keun Sohn(孫再根),Kato Akira(加藤明),Oono Kiyoharu(大野 淸春) 한국육종학회 1997 한국육종학회지 Vol.29 No.3
The purpose of this study was to detect and map quantitative trait loci(QTL) related to rice seedling growth at low temperature. The F₂ populations of 94 plants was obtained from a cross between a coldsusceptible indica cultivar ‘Dular’ and a cold-tolerant japonica cultivar ‘Toyohatamochi’. A molecular genetic map was constructed using random amplified polymorphic DNA (RAPD) markers and QTL associated with the cold tolerance of rice seedling at 18℃ was analyzed using MAPL-MQTL computer program. Analysis of RAPD patterns regarding to cold tolerance in F₂ population of Dular/Toyohatamochi cross indicated that two RAPD markers 20 and 29 were located with 21 cM apart on chromosome No. 5 and QTL related to seedling growth at 18℃ was closely associated with these DNA markers. The association of the RAPD markers on chromosome 5 with the seedling growth at 18℃ was observed in a separate analysis using F₃ plants of the cold-susceptible indica cultivar ‘K-sen 4’ and the cold-tolerant javanica cultivar ‘Silewah’.