Study on the transformation of rice ( Oryza sativa L.) Tainung 67 with Glycerol Kinase ( GK )

• Main Authors: Chang-Che Lo, 羅章哲
• Other Authors: Fure-Chyi Chen
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/38139494624739450687

碩士 === 國立屏東科技大學 === 生物科技研究所 === 94 === The purposes of this research were to study the role and effect of glycerol kinase gene (GK), which is a key enzyme of glycerol metabolism, isolated from scented rice by transforming into non-scented rice. Glycerol metabolism was reported to be involved in the synthesis of the dihydroxyacetone phosphate (DHAP), one of the probable precursor of the aromatic component 2-acetyl-1-pyrroline (2-AP) from scented rice. The GK cDNA cloned from scented O. sativa TNG71 was constructed in a binary vector and introduced into non-scented O. sativa TNG67 via Agrobacterium-mediated transformation. β-glucuronidase (GUS) reporter gene activity was detected in transformed calli and leaves of transgenic rice plantlets. Amplification of GUS gene by polymerase chain reaction (PCR) of genomic DNA and subsequent Southern-blot analysis from the leaves of transgenic rice confirmed the integration GK gene in the rice genome. Besides, different GK promoter regions were fused to GUS-reporter gene and transformed into callus of rice TNG67 for activity evalution. The results showed that GK promoter with TATA box drove stronger GUS activity than that without TATA box. GK promoters containing ABREs cis-element drove stronger GUS activity in carbon-free medium than that in other carbon source. The activity of different GK promoter regions in glycerol as carbon source was slightly lower than that in carbon-free medium. Addition of abscisic acid (ABA) to sucrose containing medium stimulated higher GUS activity driven by GK promoter with ABREs element.