摘要：水稻是人类的主要粮食之一，雄性不育系的研究对优化水稻种质资源、提高产量等有重要作用。花药是影响花粉发育的重要器官，研究花药的结构功能有助于探索花粉败育的原因，进一步认识水稻的雄性不育机理。本实验用来自于TOS17逆转座子插入的突变体ms4作为研究材料。ms4与野生型相比较，营养阶段发育正常且雌配子发育正常，但是不产生花粉，说明雄配子的发育受到了影响。本实验运用表型分析、基因鉴定、染色体DAPI染色观察、亚细胞定位、树脂切片分析以及相关基因比较等多种方法，确定该突变为MS4基因，该基因在花药中表达量极高，且不影响雄配子正常减数分裂，主要影响绒毡层的降解导致花粉无法释放而产生败育。93502

毕业论文关键词：水稻，雄性不育突变体，MS4，基因克隆，功能分析

Abstract: Rice is one of the main food for human beings and the study on the male sterile line of rice plays an important role in optimizing the rice germplasm resources and increasing yield。 Anther is an important organ that affects the development of pollen。 The study on the structure and function of anther is helpful to explore the reason of pollen abortion, and to further understand the mechanism of male sterility in rice。 In this study, the mutant ms4 was identified through a TOS17 mutant。 Compared with the wild type, the vegetative stage of Ms4 was normal and the development of female gametophyte was normal, but no pollen was produced。 In this study, we used many methods to determine the mutation gene ms4 gene such as phenotypic analysis, gene identification, chromosome DAPI staining observation, subcellular localization, resin slice analysis and related gene comparison and so on。 The expression of this gene in anther is very high, which does not affect the normal meiosis of male gametes。It mainly affects the degradation of tapetum, which leads to the release of pollen。

Keyword: rice, male sterile mutant, MS4, Gene cloning，functional analysis

目   录

1  前言 4

1。1  水稻雄性不育系简介 4

1。2  国内外研究现状、水平和发展趋势 4

1。2。1  植物有性生殖过程 4

1。2。2  花药发育的基本过程 6

1。2。3  植物花药绒毡层发育过程及其功能 8

1。2。4  绒毡层发育过程相关基因分类及其研究进展 9

2  材料与方法 11

2。1  实验材料 11

2。1。1 供试水稻亲本或品种 11

2。1。2 菌株及载体 11

2。1。3 实验试剂 11

2。2  实验方法 11

2。2。1  材料种植 11

2。2。2  表型鉴定 11

2。2。3  减数分裂染色体DAPI染色观察 12

2。2。4  亚细胞定位 12

2。2。5  树脂切片分析 13

3  结果与分析 13

3。1  MS4突变体表型分析 14

3。2  MS4突变基因的分离鉴定