【作者】 刘平；

【导师】 侯岁稳；

【作者基本信息】 兰州大学 ， 细胞生物学， 2010， 硕士

【摘要】 水稻是世界上主要的粮食作物之一,超过二分之一的人以其为主食。随着水稻全基因组测序的完成,加上各种数据库的构建和公布,同时由于水稻具有较小的基因组与禾谷类作物的共线性,成熟的遗传转化体系,使其成为单子叶的模式植物。因此其基因的功能及相互作用成为基础研究的热点和人类社会发展的要求。气孔和表皮是研究细胞分化和植物发育的模式系统。在拟南芥中,气孔和表皮的研究已经取得很大的成果,且日益白炽化,而水稻领域则方兴末艾。本实验建立了水稻种植和筛选突变体的体系,并筛选得到了4个水稻T-DNA插入的突变体,其中一个表皮平滑型(245em),一个气孔成簇型(340),一个侏儒型(245dm)和一个增高型(367)。通过初步的表型鉴定后,着重对气孔成簇型和侏儒型进行了细胞和分子水平上的分析。245em的主要表型是叶片表皮之间相互咬合的类似于拟南芥叶片表皮的突起(1obe)消失,表皮之间变的平滑,同时硅质的乳突在数量和形态上都发生了突变。245dm型的主要表型是在5叶以前植株整体很明显的矮化,叶片变小,后面的生长阶段会有一定程度的恢复,但是同样的表型在下一代会重复出现。叶片表皮细胞被压缩,部分保卫细胞母细胞不分裂成保卫细胞。245dm的的插入位点已经通过hiTAIL-PCR找到,通过半定量RT-PCR和生物信息学分析初步推定突变基因是6580。340除了叶片表皮气孔成簇以外,花的各种器官也存在位置、形态和功能上的畸变,同时花的器官有退分化然后再分化形成新植株的能力。通过southern blotting和PCR分析表明340的插入片段不是从质粒上T区的边界插入的。插入片段在左臂向V区延伸,在右臂则在T区内部断裂。更多还原

【Abstract】 Rice is a main cereal crop and staple food for over half of the world population. Rice has also become a model system of monocotyledon plants for genomic studies because of its relative small genome, mature transformation technique, the published various databases and the completion of the genomic sequencing projects of both indica and japonica subspecies.Stomata and epidermal cells were both model systems for studying the cell differentiation and plant development. In Arabidopsis, the great achievements of stomatal and epidermal development have been made. However, there was less correlative advancement in rice.In this experiment, we created the rice planting and mutant screening system, and screened four rice T-DNA insertion mutants, a smooth skin type (245em), a stomatal clusters type (340), a dwarf-type (245dm) and a higher type (367). After the initial phenotypic identification,340 and 245dm were chose to analyze in cellular and molecular level.The primary phenotype of 245em was that the lobes of leaf epidermis (similar to Arabidopsis lobes) disappeared and the epidermal cells became smooth. And the number and morphology of silica papillae were changed.The central phenotype of 245dm was that the plant was obvious dwarfing with smaller leaves before the 5th leaves stage. After the stage, the mutant phenotype of 245dm could recover to a certain extent. However, the same mutant phenotype of 245dm in the next generation would appear again. And leaf epidermal cells were compressed and part of guard mother cells (GMCs) did not divide to form guard cells. The T-DNA insertion site of 245dm has been found by hiTAIL-PCR, and 6580 was the presumption mutant gene by semi-quantitative RT-PCR and bioinformatics analysis.In 340, the leaf stomata clustered and the position, shape and function of various flower organs distorted. The differentiation of flower organs has degenerated and these mutant flowers could form new plants directly. The T-DNA inserted fragment in 340 was detected by southern blotting and PCR analysis, and found that the inserted fragment was not from the border of T-area of the DS-GUS plasmid. The left arm of inserted fragment extended to the V area of plasmid and the T area fractured within the right arm.更多还原