【作者】 滕蕾；

【导师】 杨毅；

【作者基本信息】 四川大学 ， 遗传学， 2006， 硕士

【摘要】 Fibrillin是一类由核基因编码，与质体相关的脂连接蛋白，广泛存在于单子叶和双子叶植物中。在逆境条件下，如强光、寒冷和干旱环境，该类蛋白大量积累，说明Fibrillin在生物胁迫下发挥重要功能。在拟南芥中已发现Fibrillin是ABI2(ABA insensitive)相互作用蛋白质，其合成也依赖于ABI1，而ABI1和ABI2是脱落酸(ABA)信号传导过程中的关键因子，推测Fibrillin作为负调控因子，参与到ABA信号传导途径。同时Fibrillin与油质体相关，强光下对光合系统Ⅱ有保护作用，则该基因在油菜中的过量表达可能有利于提高油菜的抗逆能力和产油量。因此研究Fibrillin在油菜中的表达具有重要的理论意义和实践意义。 在本实验中，我们将拟南芥Fibrillin操纵子连接到带有荧光素酶(LUC)报告基因的pBI221质粒上，通过聚乙二醇(PEG)导入甘蓝型油菜蜀杂9号原生质体中进行瞬时表达，检测报告基因荧光素酶的表达量，初步得到油菜原生质体表达系统。加入一定量的脱落酸(ABA)后，发现报告基因的表达量有上升趋势，说明ABA能诱导报告基因的表达，拟南芥Fibrillin操纵子在甘蓝型油菜原生质体中对ABA的诱导敏感。 为了进一步验证Fibrillin与ABA之间的关系，我们把Fibrillin操纵子和Fibrillin cDNA分别连接到带有葡萄糖苷酶(GUS)报告基因的pBI121质粒上，经农杆菌浸染甘蓝型油菜84100-18的下胚轴，以期获得稳定表达的转基因植株。我们根据材料特点，优化了预培养、农杆菌浸染、共培养、诱导愈伤、诱导芽、生根等组织培养的各个步骤，最终经卡那霉素筛选获更多还原

【Abstract】 Plant fibrillin is a nuclear-encoded plastid lipid-associated protein that ubiquitously exists in both monocotyledonous and dicotyledonous plants. Protein accumulation occurs under environment stress such as high light, cold and drought, showing that fibrillin functions in adapting to abiotic stress and may play an important role in plant growth and development. In Arabidopsis thaliana, it has been proved that fibrillin protein interacts with ABI2(ABA insensitive) and its synthesis is dependent on ABI1, both of which are key factors in ABA signal transduction, indicating fibrillin roles in ABA signal transduction as a negative factor. In addition, fibrillin is known to relate to oleosome and to participate in PSII protection against illumination, cluing that its overexpression in Brassica napus will help to improve plants’ oil production and adaptability to adverse circumstances. So it is very important in theory and application to research fibrillin’s expression and function in Brassica napus.In our experments, arabidopsis fibrillin promoter was constructed in pBI221with luciferase reporter gene and transferred to protoplasts of restorer line of84100-18, male parent of a new sort of hybrid rapeseed "SHUZH A No.9", viaPEG-mediated method, showing a transient expression. After adding 30μM ABA,we found that the expression was upregulated, indicating fibrillin is sensitive to更多还原