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摘要水稻是我国重要的粮食经济作物，叶片是植物进行光合作用的主要场所。叶色的改变将影响水稻的产量，因此有关水稻叶色突变体的研究具有重要意义。在实验室条件下，通过EMS诱变籼稻品种“蜀恢”，得到一个在整个生长期表现为淡黄叶表型的叶色突变体，称其为淡黄叶突变体yl1。该突变体与野生型相比，叶绿素a、叶绿素b和总叶绿素含量明显降低，叶绿体发育受损，体积小、类囊体结构松散。净光合速率(Pn)、蒸腾速率(Tr)、和气孔导度(Gs)等光合指标较野生型也显著下降。以yl1突变体为母本，粳稻品种日本晴为父本建构F2定位群体，确定突变体由隐性单基因控制，通过图位克隆将YL1基因定位于水稻3号染色体SSR标记YP2033和YP1963之间。通过生物信息学方法分析得出定位于叶绿体的候选基因，对比野生型和突变型的碱基序列，结果显示该基因全长600bp，第322位碱基由C突变为T，导致脯氨酸变为丝氨酸。运用多种软件对YL1蛋白结构域及二级结构、蛋白理化性质、定位、特异性表达情况、磷酸化和糖基化位点等进行生物信息学分析, 帮助我们预测YL1基因功能。47915
Rice is one of the most important food and economic crop. Leaves are main organs where photosynthesis is being conducted. The change of leaves color will influence the output of rice significantly, so it seems really important to do some research for leaf-color mutant. we treated the indica rice “Shuhui” with Ethylmethan Sulphonate (EMS) under laboratory conditions, a leaf-color mutant displaying distinct yellow leaf phenotype throughout development was identified from the progeny, called yellow-leaf mutant yl1. Compared with wild type, the content of Chlorophyll a, Chlorophyll b and total chlorophyll are obviously decreased. Chloroplast development is not very well and just has small volume, the thylakoid structure is loose. Photosynthetic indicators such as net photosynthetic rate (Pn), transpiration rate (Tr) and stomatal conductance (Gs) are significantly reduced. We crossed mutant yl1 with japonica cultivar Nipponbare to establish a F2 population, it was determined that the phenotype of mutant is controlled by single stealth genes. We detected that YL1 is located on chromosome 3 between SSR maker YP2033 and YP1963 by map-base cloning. By method of bioinformatics, we find the candidate genes which located in the chloroplast. Compared the base sequence of wild type and mutant, it showed that the candidate genes has 600 bp with a single C to T conversion at the 322nt of coding region, which caused the amino acid conversion of pro to ser. The analysis of bioinformatics such as YL1 protein structure domain and secondary structure, physical and chemical properties of protein, positioning, specific expression, phosphorylation and glycosylation sites will be done by using a variety of software, witch can help us to reveal the function of YL1 genes.
毕业论文关键词：水稻；叶色突变体；图位克隆；功能分析
Keyword: rice, mutation of leaf color, map-base cloning, function analysis
目 录
摘 要 2
0 引言 4
1 材料与方法 4
1.1 水稻材料 4
1.2 实验方法 4
1.2.1 叶绿色含量的测定 4
1.2.2 叶绿体发育的电镜观察 5
1.2.3 定位群体的构建 5
1.2.4 图位克隆 5
1.2.5 水稻叶片DNA的提取