Antisense gibberellin 3β-hydroxylase발현 형질전환벼

Abstract

GA 생합성에 결정적 역할을 하는 GA 3<TEX>$\beta$</TEX>-hydroxylase를 pIG121-Hm 벡터에 GUS유전자를 빼고 antisense로 클로닝하여 이를 동진벼에 도입한 결과 17개체의 절간신장이 억제된 형질전환한 식물체를 얻을 수 있었다. 일반 재배 동진벼를 대조군으로 하여 비교하였을때 antisense GA 3<TEX>$\beta$</TEX>-hydroxylase 유전자가 형질 도입된 식물체의 획득형질은 평균적으로 대조군에 비해 절간 신장의 억제가 확인되었다. 절간신장의 억제가 보인 개체의 엽육조직을 co취하여 Southen blot hybridization분석 결과 3개의 line에서 모두 single copy로 도입된 것으로 나타났다. 이로써<TEX>$T_o$</TEX> 식물체 내에 antisense GA 3<TEX>$\beta$</TEX>-hydroxylase 유전자를 내포하고 있는 것으로 확인되었다. 이것은 antisense GA 3<TEX>$\beta$</TEX>-hydroxylase 유전자가 생체내에서 직접 또는 간접적으로 GA 3<TEX>$\beta$</TEX>-hydroxylase유전자의 발현에 관여한 것이라 사료되어진다. During plant development, active gibberellins (GAs) control many aspects of plant growth and development including seed germination, stem elongation, flower induction, anther development and seed growth. To understand the biosynthesis and functional role of active GAs in high plants, this study investigated GA 3<TEX>$\beta$</TEX>-hydroxylase gene en-coding <TEX>$GA_1$</TEX> and<TEX>$GA_4$</TEX> catalizing last step in GA biosynthetic pathway. The antisense GA 3<TEX>$\beta$</TEX>-hydroxylase gene was inserted into expression vector, pIG121-Hm. Calli derived from mature seeds of rice (Oryza satiiva L. cv. Donjinbyeo) were co-cultivated with Agrohacterium tumefaciens EHA101 earring a pIG121-Hm containing hygromycin resistance (<TEX>$Hyg^r$</TEX>) and antisense GA 3<TEX>$\beta$</TEX>-hydroxylase gene. Seventeen transgenic plants obtained inhibiting GA 3<TEX>$\beta$</TEX>-hydroxylase. Transgenic plants had shorter plant height more than that of the Dongjinbyeo. Stable integration of antisense GA 3<TEX>$\beta$</TEX>-hydroxylase gene was confirmed by polymerase chain reaction of genomic DNA isolated from the leaf organs of the <TEX>$T_o$</TEX> generation.