番茄EMS突变体库的构建及叶色黄化突变体的分析

发布时间：2018-05-10 14:48

本文选题：番茄 + EMS　；参考：《沈阳农业大学》2017年硕士论文

【摘要】：番茄(Solanum lycopersicum)作为目前世界上栽培面积最大、产量最高的蔬菜,既是人们生活中必不可少的新鲜蔬菜,也是重要的食品加工原料。在植物研究中,番茄作为重要的模式植物,在植物科学研究中也具有重要地位。突变体是新品种选育和研究功能基因组学良好的试材,而目前番茄的突变体库还相对较少。因此,本论文以测序番茄品种'Heinz 1706'为试材,采用不同EMS诱变剂处理番茄种子,在后代筛选各种变异植株,获得稳定变异株系,旨在建立番茄EMS突变体库,并对一个叶色黄化的突变株系进行了分析。论文的主要研究结果如下:1.以番茄'Heinz 1706'种子为试材,采用不同浓度EMS(0.30%、0.5%、0.70%、0.9%、1.0%、1.3%、1.5%、2.0%)诱变剂处理8h和12h,结果表明,随着EMS诱变浓度的升高与处理时间的延长,种子发芽率逐渐降低。高浓度处理显著降低了植株的结实率,影响了植株正常的生长发育。从植株突变率与生长发育状况综合考虑,最终确定了适宜的处理方法为1%浓度的EMS诱变种子12h.2.通过统计M2群体7864个单株的生长发育特性,获得了 553株变异单株,得到了 1160个叶形态、叶色、花形态、果实大小、株型等变异性状。总的单株变异数为7.00%,其中324株是有多个变异表型,229株只有单一变异表型。叶、花、果实、植株的表型变异株数分别为375、183、196和460,变异频率分别是4.77%、2.33%、2.49%、5.85%。对变异植株进行留种,初步构建了番茄'Heinz 1706'的EMS突变体库。3.从M2代群体中获得一个编号为55的叶色黄化突变体(Y55),在整个植物生长期间表现出明显的黄色叶表型,其生长势比野生型要弱,但能正常开花结实;测定Y55突变体叶片的色素及其前体含量,结果表明Y55的叶绿素a、叶绿素b、类胡萝卜素、总叶绿素含量以及叶绿素a/b比值均显著低于野生型,同时,Y55叶绿素合成的前体物质含量均较野生型显著降低,由此表明Y55叶绿素含量的下降可能也是由于整个叶绿素生物合成途径的合成能力下降造成的;根据扫描电镜和透射电镜的观察,发现Y55叶片下表皮保卫细胞面积、气孔面积及茸毛数均显著低于野生型,叶绿体发育结构受损;对Y55叶片光合和荧光参数分析表明,Y55光合作用显著降低,光合效率及耐光抑制能力均低于野生型;遗传分析表明该黄叶表型由单个隐形基因控制。综上,本论文筛选建立了测序番茄品种'Heinz 1706'EMS诱变的处理体系,初步建立了突变体库,并对叶色黄化突变体Y55初步形态结构、生理参数和叶色性状遗传规律进行分析,为进一步研究相关基因功能奠定了坚实的基础。
[Abstract]:Tomato Solanum lycopersicum (Solanum lycopersicum), as the largest cultivated area and the highest yield vegetable in the world, is not only a necessary fresh vegetable in people's life, but also an important raw material for food processing. Tomato, as an important model plant in plant research, also plays an important role in plant science. The mutants are good materials for breeding and studying functional genomics of new varieties, but the mutants library of tomato is relatively few at present. Therefore, using sequenced tomato variety Heinz 1706' as the test material, different EMS mutagens were used to treat tomato seeds. In order to establish the EMS mutant library of tomato, a variety of mutated plants were screened out in the progeny of tomato, and stable mutant lines were obtained. A leaf yellowing mutant strain was analyzed. The main results of this paper are as follows: 1. The seeds of Heinz 1706' were treated with different concentrations of EMS0.300.0.700.70 and 0.70, respectively. The seeds of Heinz 1706' were treated with mutagens for 8 hours and 12 hours. The results showed that the seed germination rate decreased gradually with the increase of EMS concentration and the prolongation of treatment time. High concentration significantly reduced the seed setting rate and affected the normal growth and development of plants. Considering the plant mutation rate and growth and development, the suitable treatment method was determined as 1% EMS mutagenesis seed 12h.2. Through statistics of growth and development characteristics of 7864 individual plants in M2 population, 553 variant individual plants were obtained, and 1160 leaf morphology, leaf color, flower morphology, fruit size and plant type were obtained. The total variation per plant was 7.00, of which 324 had multiple variation phenotypes and 229 had a single variant phenotype. The phenotypic variation of leaves, flowers, fruits and plants were 375183196 and 460 respectively, and the frequency of variation was 4.77 and 2.33, respectively. The EMS mutant library of Heinz 1706 'was constructed. A leaf color yellowish mutant Y5555 was obtained from M2 population. It showed a yellow leaf phenotype during the whole plant growth period, and its growth potential was weaker than that of wild type, but it could bloom and fruiting normally. The results showed that the chlorophyll a, chlorophyll b, carotenoid, total chlorophyll content and chlorophyll a / b ratio of Y55 mutant were significantly lower than those of wild type. At the same time, the content of precursor substances of Y55 chlorophyll synthesis was significantly lower than that of wild type, which indicated that the decrease of Y55 chlorophyll content may also be due to the decrease of the synthesis ability of the whole chlorophyll biosynthesis pathway. According to the observation of scanning electron microscope and transmission electron microscope, it was found that the area of epidermal guard cells, stomatal area and hairy number of Y55 leaves were significantly lower than those of wild type, and chloroplast development structure was damaged. The photosynthetic and fluorescence parameters of Y55 leaves were analyzed. The photosynthetic efficiency and light tolerance of Y55 were lower than that of wild type, and the phenotype of yellow leaf was controlled by a single invisible gene. In this paper, the mutagenesis system of sequenced tomato variety Heinz 1706'EMS was established, and the mutant library was established. The preliminary morphological structure, physiological parameters and genetic rules of leaf color traits of Y55 were analyzed. It lays a solid foundation for further study of related gene function.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S641.2

【相似文献】