兰州大学机构库 >生命科学学院
拟南芥ckrc突变体的筛选和表型研究
Alternative TitleIdentification and phenotype characterization of ckrc mutants in Arabidopsis thaliana
张天仔
Thesis Advisor郭光沁
2013-05-26
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword玉米素 生长素 突变体筛选 zb11
Abstract生长素作为植物体内重要的激素,调控着植物生长和发育等重要过程,如影响细胞的伸长、分裂和分化、促进根茎叶的生长、维管组织分化、顶端优势和向地性以及向光反应等。 生长素的合成是非常复杂的,包括色氨酸依赖途径和色氨酸非依赖途径,其中有很多步骤是未知的。有鉴于此,通过本实验室建立的生长素代谢缺陷突变体筛选体系,即ckrc(cytokinin induced root curling)对T-DNA插入突变体库进行筛选。cs22830含有37,800个lines库,共筛选257,500粒种子,得到209株在0.1μM玉米素(trans-zeatin,tZ)培养基上具有根卷曲表型的苗,对其进行复筛,发现有10株在tZ培养基上具有根卷曲生长的表型。对此10个突变体进行生化互补实验,并归类。其中pin2类突变体有两个t1和t2;aux1类突变体有四个t5、t6、t7和t8;wei2/wei7类突变体有4个t3、t4、t9和t10。 对本实验室筛选到的另一个根卷曲的突变体zb11进行系统的遗传表型分析。显示zb11为单位点隐性突变,主根变短以及根向重力性缺失等低生长素表型,且这些缺陷可被外源IAA(Indole-3-acetic acid)恢复。此外,tZ培养基上卷曲生长的根能被外源添加的低浓度生长素IAA、NAA和2, 4-D互补。zb11的内源生长素含量测定显示其游离态IAA以及结合态IAA均降低,表明zb11是一个低生长素缺陷突变体。
Other AbstractAuxin plays an important role in plant growth and development, including elongation, division and differentiation of cells, growth of roots and leaves, differentiation of vascular tissue, dominance and photoisomerization of plant. There are multiple pathways for auxin biosynthesis and metabolism, forming a complex web, for which we are still far from forming a clear picture. Based on our previous works on ckrc1-1 (CKRC1 encodes tryptophan aminotransferase (TAA1) for IAA biothynthesis), the aim of this study to isolate more Arabidopsis ckrc mutants that are likely to have low auxin (IAA) levels in roots, on which further molecular genetic and biochemical studies should uncover important genes in controlling auxin biosynthesis and metabolism. 257,500 seeds were screened and there are 209 seedlings with curling root phenotype when grown in MS medium containing 0.1μM tZ. 10 of those lines showed root curling in the second screening. Biochemical complementation experiment showed that there are two pin-like mutants t1 and t2, four aux-like mutants t5, t6, t7 and t8, four wei2/wei7-like mutants t3, t4, t9 and t10 in the 10 mutants. Studies on another mutant zb11 showed that this mutant had the phenotype of auxin deficient such as its relative short primary root and weak gravitropism in its roots, which can be rescued by exogenous Indole-3-acetic acid (IAA). Moreover, the root curling phenotype of zb11 could be complemented by low concentrations of IAA, NAA or 2,4-D. GC-MS results confirmed that the endogenous IAA level in zb11 mutant was significantly reduced, indicating that zb11 is an auxin deficient mutant.
URL查看原文
Language中文
Document Type学位论文
Identifierhttp://ir.lzu.edu.cn/handle/262010/220722
Collection生命科学学院
Recommended Citation
GB/T 7714
张天仔. 拟南芥ckrc突变体的筛选和表型研究[D]. 兰州. 兰州大学,2013.
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