Functional Analysis of Rice Oleosin by RNA Interference

• Main Authors: Anda Chou, 周郁如
• Other Authors: Liang-Jwu Chen
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/89086881756506935700

碩士 === 國立中興大學 === 分子生物學研究所 === 93 === Two rice oleosin isoforms, with molecular mass of 16 kD (Os701) and 18 kD (Os721), have been confirmed to be embedded in the rice oil bodies. To understand the role these two oleosin isoforms play in forming the oil body, we try to knock out the expression of each individual oleosin using RNA interference approaches. In this study, the Os701 gene was the first target to be knocked out. A construct that can potentially express double-stranded RNA in the transgenic rice were obtained and used to transform rice plants. Six transgenic lines with significantly reduced or lack of the expression of 16 kD oleosin were confirmed by Western blot using total extractable seed proteins. The observable phenotype of transgenic plants showed no difference from those of the host plants. However, a significant reduction of the oil body content during fractionation and a significant enlarged oil bodies in the embryonic cells were observed in transgenic rice. In the oil bodies, these two oleosin isoforms remained at an almost 1:1 ratio, but with significantly reduced total amount of oleosin. The rest of the large amount of 18 kD oleosin disappeared from the mature oil body. Current investigation of the distribution of the rest of the 18 kD oleosin in these transgenic plants are underway. These preliminary observations suggest that the 16 kD oleosin isoforms may play a more important role in forming the oil bodies.