| Proanthocyanidin (PC), or condensed tannins, belongs to flavonoids. It is a type of substance with antioxidation,scavenge free radical and Anti-tumor and a series of biological activities. As one of the world's most popular 10 medicinal plants, PC has been widely used in medicine and cosmetics.Rice and grapes have similar metabolic pathways, with a common precursor substances– anthocyanin, anthocyanin reductase (ANR) converts anthocyanins to proanthocyanidins in grape, but no ANR in rice. This study attempt to genetically engineering to co-express ANR gene from grape induce rice. The expression in the embryos and aleurone layer of transgenic rice should rise the content of proanthocyanidin. The main results include:1.The plant expression vector pCDMAR-ANR-Hyg was constructed, which contained two spereate T-DNA regions, One carried the selectable marker gene (Hyg), the other carried the ANR gene. The ANR gene was driven by rice seed specific Oleosin promoter, two sides of the ANR gene cassette with two direct repeat Rb7 MARs of tobacco, in order to enhance expression of the ANR gene.2. The plant expression vector was transformed to the embryonic callus tissue of Taijing 9 through Agrobacterium-mediated method. We obtained five transgenic plants, two of which were positive by PCR testing.3. With the regenetation system in our lab, comparative analysis of different types of black rice, methods of sterilizing and different mediums on callus regeneration ability of rice. The main results showed that NB and N6D medium are better for longjin 1, riben and 531. The plant expression vector pCDMAR-ANR-Hyg transformed to the embryonic callus tissue of 531 through Agrobacterium-mediated method. We obtained resistance embryonic callus tissue. |
