甜菜育性相关基因蛋白差异表达分析

发布时间：2018-05-29 01:48

本文选题：甜菜 + 细胞质雄性不育　；参考：《内蒙古农业大学》2016年硕士论文

【摘要】：为了更好的探索甜菜不育系质核基因在蛋白调控上的相关关系,本研究通过对内蒙古农牧科学院甜菜所提供的同一育种背景下拟选为不育系的未成系甜菜植株及成系的甜菜不育系及对应保持系(N9849和960767)材料进行田间花粉调查,并对花粉调查结果中的有粉植株进行VNTR分子鉴定获得的甜菜拟选为不育系中的S型胞质有粉材料,S型胞质无粉材料(不育系)及其对应保持系利用蛋白质双向凝胶技术获得对应的双向电泳图谱。进而对获得的双向电泳图谱进行图谱软件分析获得特异差异蛋白点。得到结果及结论如下。1、通过对91份供试材料的田间调查,除41份未抽薹,4份未出苗外获得25份有粉材料,21份无粉材料；利用VNTR微卫星分子分子标记鉴定技术对25份有粉材料进行鉴定,获得S型细胞质材料5份。2、对甜菜叶片及花器蛋白进行TCA-丙酮沉淀法和丙酮酚法两种蛋白提取方法的比较、优化得到适合甜菜叶片及花器蛋白蛋白提取并满足双向凝胶电泳差异分析的TCA-丙酮沉淀蛋白提取方法。3、优化了甜菜蛋白质双向电泳中等电聚焦过程技术参数,得到清晰的双向蛋白图谱。4、以不育系及保持系花期叶片为研究材料,进行蛋白质双向电泳研究获得385个蛋白点,差异点12个,其中不育系中特异表达的7个,保持系中特异表达5个；以花器为研究材料,获得差异蛋白点23个,其中不育系中特异表达的15个,保持系中特异表达8个。5、对实验中筛选出的各S型胞质有粉材料,同参考不育系对比发现差异点数在除不育系所特有差异点外,各材料所特有差异点分别为,5、4、6、8、11。
[Abstract]:In order to better explore the relationship between cytoplasmic nuclear gene and protein regulation in sugarbeet sterile line, In this study, field pollen investigation was carried out on sugarbeet plants selected as sterile lines under the same breeding background provided by Sugarbeet of Inner Mongolia Academy of Agriculture and Animal Husbandry, as well as adult sugarbeet sterile lines and maintainer lines N9849 and 960767. The S type cytoplasmic pollen material (CMS line) of S type cytoplasmic powder was selected by VNTR molecular identification of powdered plants in pollen investigation results and its application of protein bidirectional coagulation to the maintainer line. The corresponding two-dimensional electrophoretic patterns were obtained by gel technique. Then, the specific differential protein spots were obtained by the analysis of the two-dimensional electrophoresis map software. The results and conclusions are as follows: 1. Through the field investigation of 91 tested materials, 25 powdery materials were obtained except 41 without bolting and 4 without sprouting. 25 powdered materials were identified by VNTR microsatellite molecular marker technique, and 5 samples of S-type cytoplasm were obtained. The extraction methods of sugar beet leaves and floral proteins were compared by TCA-acetone precipitation method and pyruvol method. The extraction method of TCA-acetone precipitated protein suitable for extracting sugar beet leaves and floral proteins and satisfying the differential analysis of two-dimensional gel electrophoresis was obtained. The technical parameters of medium electric focusing process of 2-D electrophoretic electrophoresis of beet protein were optimized. A clear two-dimensional protein map of .4 was obtained, and 385 protein spots were obtained by using leaves of male sterile lines and maintainer lines at flowering stage, with 12 differentially expressed spots, among which 7 of them were specifically expressed in male sterile lines. Using flower organ as the research material, 23 differentially expressed proteins were obtained, of which 15 were specifically expressed in male sterile lines and 8. 5 in maintainer lines. In comparison with the reference male sterile lines, it was found that the difference points of the number of different points except for the specific points of the sterile lines were respectively the different points of 5, 5, 4, 4, 5, 5, 5, 4, 5, 5, 4, 5, 5, 4, 4
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S566.3

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