摘要:ABC1激酶是原核和真核生物中普遍存在的一类非典型蛋白激酶,在原核生物和真核线粒体中主要调控泛醌的生物合成。近来研究表明,拟南芥叶绿体ABC1激酶调控类异戊二烯代谢、铁离子分布和光氧化胁迫的应答,而重要作物如水稻ABC1激酶的功能尚不明确。本研究利用T-DNA插入突变体对水稻叶绿体ABC1K6的分子功能进行了鉴定。结果表明,osabc1k6突变体株高略矮于野生型,叶绿素含量显著下降,而类胡萝卜素含量无显著差异。甲基紫精处理表明,突变体和野生型表型及丙二醛含量无显著差异,说明OsABC1K6基因不直接参与氧化胁迫的防御。实时定量PCR结果显示,osabc1k6基因突变影响了叶绿体类异戊二烯合成相关基因的表达。这些结果为进一步阐明OsABC1K6的分子功能奠定基础。54343
毕业论文关键词:水稻,ABC1基因,T-DNA插入突变体,类异戊二烯代谢
Abstract: ABC1 kinases are atypical protein kinases that are widely found in prokaryotes and eukaryotes. They play essential roles in regulating ubiquinone biosynthesis in prokaryotes and eukaryotic mitochondria. Recent studies in Arabidopsis showed that chloroplast ABC1 kinases modulate prenyllipid metabolism, iron distribution and photooxidative stress response. However, their roles in important crops such as rice remain illusive. In this study, a rice ABC1 kinase ABC1K6 was characterized using T-DNA insertion mutants. The plant height and chlorophyll contents were lower in osabc1k6 mutants than in wild type plants; however, the carotenoid content was not significantly different. The mutant phenotype and malondialdehyde content were also the same under methyl viologen treatment, suggesting that OsABC1K6 did not directly participate in oxidative stress response. Real-time PCR analysis showed that expression of the chloroplast prenyllipid biosynthetic pathway genes was affected in osabc1k6 mutants. These results will lay a solid foundation for further functional analysis of the OsABC1K6 kinase.
Keywords: rice, ABC1 gene, T-DNA insertion mutant, prenyllipid metabolism
目 录
1 前言 4
2 材料与方法 5
2.1 植物材料和处理 5
2.2 色素含量的测定方法 5
2.3 丙二醛(MDA)含量的测定 5
2.4 RNA的提取和实时定量PCR 6
3 结果与分析 6
3.1 纯合突变体鉴定 6
3.2 水稻abc1k6突变体对氧化胁迫不敏感 7
3.3 水稻ABC1K6基因突变影响了叶绿体类异戊二烯合成相关基因的表达 8
讨 论 10
参考文献 12
致 谢 14
1 前言
生物体存在多种复杂的代谢调节方式,而调节生物体信息转导最普遍的方式是蛋白激酶催化蛋白质磷酸化以及蛋白磷酸酶催化蛋白质去磷酸化。这一过程几乎涉及所有生物的生理调节过程,比如糖代谢、植物的光合作用、细胞的生长与发育、基因的表达等方面[1]。蛋白激酶超家族能够细分为真核生物蛋白激酶和非典型蛋白激酶,后者主要分布在原核生物中[2]。与典型蛋白激酶相比,非典型蛋白激酶缺少HMM结构域,但都含有250-300个氨基酸的催化功能域[2,3]。在植物中,蛋白激酶的研究目前主要集中在分裂原激活蛋白激酶家族(MAPK)和钙依赖蛋白激酶家族(CDPK)等真核蛋白激酶家族上。 水稻ABC1激酶ABC1K6的突变体分析:http://www.youerw.com/shengwu/lunwen_58546.html