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水稻种子品质性状相关突变体的筛选与鉴定

Screening and Identification of Seed Quality Trait Mutants in Rice(Oryza Sativa, L.)

【作者】 王健

【导师】 万建民;

【作者基本信息】 南京农业大学 , 作物学, 2013, 硕士

【摘要】 水稻是世界上最主要的粮食作物之一,水稻基因功能的研究已经在科研领域和生活领域起着至关重要的作用。构建水稻突变体材料是进行水稻基因研究直接有效的途径。而运用物理辐射以及化学药剂诱变的方法,以其高于自然诱变的突变频率,可以进行大量有目的的筛选,已经成为了实验室构建材料的最基本方法。本研究通过对N22、93-11、滇粳优1号三种常规水稻,进行物理射线诱变和化学MNU诱变处理,得到了大量诱变的M1代种子。种植后进行田间的突变性状的筛选,共筛选农艺性状突变体材料25份,以及收获后的种子性状的考察,构建了一批水稻基因研究的突变体材料,为在水稻功能基因的研究和水稻遗传等方面提供了有利的材料。对收获的诱变M2种子,进行脱壳观察,主要观察水稻胚乳的突变性状,从中筛选出垩白,粉质等相关性状的材料,着重考察水稻胚乳性状的差异。共筛选出了淀粉突变体752份。其中,N22,283份;9311,176份;滇粳优1号,293份。种子筛选后,对筛选出的粉质突变体进行进行SDS-PAGE电泳,观察主要蛋白含量的带型变化。通过大量SDS-PAGE电泳分析,最终筛选出水稻谷蛋白57H突变体13分,其中N227份、93112份滇粳优1号4份。对其中来源于9311的纯合的水稻蛋白突变体gpa4-1,突变体表型为叶尖衰老型黄化,胚乳粉质,谷蛋白57kD前体增加。与N22配组后,利用F2群体进行了初定位分析,初步定位于第三染色体的17.5cM区间内,是一个全新的谷蛋白前体增加突变体。本实验诱变方法获得的谷蛋白突变体有可能在谷蛋白的研究中发现新的基因,为实验室今后的谷蛋白研究打下了坚实的基础。

【Abstract】 Rice has now become one of the world’s most important food crops. As a model crop, it also provides the basis for monocots researching. Rice gene function studies play vital roles in the areas of both scientific research and life. Building rice mutants is a direct and effective way for researching rice genes. Meanwhile, using physical radiation and chemical mutagenesis can improve mutation frequency which is much higher than the natural mutation, and also can conduct a lot of purposeful selections. These establish the foundation of building materials in the laboratory.Through physical-ray mutagenesis and chemical mutagenesis MNU treatment, we get a lot of mutagenesis M1generation seeds of N22,93-11and Dianjingyou1that are three conventional rice species.Through screening mutant traits in the field, as well as examining seed traits after harvesting. The result is mutant materials25copies about agronomic traits. It builds a batch of rice mutant materials for the research of functional genes and provides favorable materials in the research of functional genes and rice genetics.After harvesting mutagenic M2seeds and peeled observation that is the main observation of rice endosperm mutants’ traits, we choose the chalky, opaque materials and other related traits. The result is mutant materials752copies about the endosperm. N22,283copies;9311,196copies; Dianjingyou1,293copies.After seed selection, we determine the protein content change of being selected starch mutants, namely conduct the SDS-PAGE electrophoresis. We screen glutelin and prolamin mutants and focus on rice protein synthesis pathways. After numerous generations of validation, we eventually eliminate the effects of environmental factors and find genetically stable protein mutant. Through a large number of SDS-PAGE electrophoresis analysis, we finally select homozygous mutant rice protein gpa4-1.The mutation phenotype are etiolating in the tip of leaves, fluory endosperm, and glutelin precursor increase. After hybridization with the N22, an F2population was constructed for positioning cloning. This gene was initially located in a17.5cM interval on the third chromosome. Genes predicted in the17.5cM interval, have not been reported before, suggesting gpa4-1is a novel glutelin precursor accumulation mutant. The laboratory may find new materials.

【关键词】 水稻诱变MNU处理突变体
【Key words】 Rice (Oryza sativa L.)MutagenesisMNU treatmentMutant
  • 【分类号】S511
  • 【被引频次】2
  • 【下载频次】340
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