不同苜蓿材料产量、品质、耐盐性评价及遗传多样性分析

发布时间：2018-06-08 13:23

本文选题：苜蓿 + 种质资源　；参考：《内蒙古农业大学》2017年博士论文

【摘要】：本研究以收集自国内外93份苜蓿(Medicago)种质为试材,在田间和室内对其生物学特性、产量、品质及分子水平的遗传多样性和蛋白质组的差异表达等进行了全面的分析研究,并对10份野生苜蓿材料进行耐盐性评价,旨在优化种质材料鉴定评价的相关研究方法,评价相关形态重要性及生理生化指标的优劣,并最终筛选优异种质材料,以为苜蓿新品种的选育提供素材与依据。试验数据经相关分析、主成分分析、通径分析以及加权隶属函数法和灰色关联度分析后,得出如下结果:(1)各农艺性状中,产量与茎粗和株高间相关性较大,相关系数分别为0.39和0.30,均呈极显著正相关。灰色关联度分析结果显示,各性状与产量的关联度范围为0.72～0.84,从大到小的排序为:株高干鲜比株丛茎叶比分枝数生长速度植株数茎粗;供试材料与标准品种之间关联度范围为0.49～0.66,当遗传距离为7时,93份材料聚为4类,其中表现最好的一类材料有4个,包括M1-5(赛特)、M1-7(West Blend)、M1-54(中苜 2 号)、M1-86(步行者 3 级)。(2)30对SSR标记对30个国内外苜蓿品种共120个基因型进行研究,共检测到565个等位基因,平均每个位点扩增得到18.83个等位基因,多态性信息含量(PIC)为0.84,且所选SSR标记的多态性信息含量均在0.6以上。120个苜蓿基因型经Structure分析处理划分为2个种群,其中种群Ⅰ包括了中国大部分的苜蓿材料,以及其他9个国家的共96个基因型,种群Ⅱ的24个基因型来自于6个国家,地理区域分别为亚洲、欧洲、美洲、大洋洲。这两个种群的遗传组成较复杂,可分别被进一步划分为2个和4个亚群,其中惊喜苜蓿在亚群划分中,与中苜2号苜蓿遗传差异较大。(3)耐盐性鉴定中,与苜蓿种子萌发期耐盐性关联度较大的指标为幼苗干重和发芽率,分别为0.752和0.745;与幼苗期耐盐性关联度较大的指标为叶片含水量和叶绿素含量,分别为0.741和0.660。耐盐性较强的材料是M3-8(来源于内蒙古通辽市大青山扁蓿豆)。(4)新品系鉴定试验中,总产量最高的材料是M4-5(新品系1),为55.92t/hm2。相关分析和通径分析结果均表明,第一茬草对总干草产量的贡献率最大。供试年中新品系1和2的的相对饲用价值较低,分别位于第12名和第11名。惊喜和草原3号苜蓿的产量和相对饲用价值均处于前5名,为高产优质材料。(5)用TMT技术对叶型差异较大的惊喜苜蓿叶片进行蛋白质组分析,鉴定出在0.05水平下,差异大于1.2倍的蛋白共53个,其中三叶型苜蓿叶片中表达上调29个,下调24个。GO注释结果中,40%的差异蛋白参与生物过程,43%参与分子功能,17%参与细胞组成。其中多叶型苜蓿中表达较高的差异蛋白参与的主要功能有细胞分化、前体代谢物和能量以及光合作用,这些生理过程都有利于植物生成新生组织,促进叶片生长。KEGG通路注释中,差异蛋白较多的前5位通路为碳代谢,剪接体,氨基酸的生物合成,内吞作用以及淀粉和蔗糖代谢,其中多叶型苜蓿中表达较多的蛋白主要参与了前4个通路,使多叶型苜蓿的碳代谢增强,促进RNA成熟和增加氨基酸的合成,有利于植物积累有机化合物,促进细胞代谢生长,从而增加产量。最终将中苜2号、新品系1号、惊喜和草原3号苜蓿以及通辽市大青山野生扁蓿豆材料作为推荐新品种选育原材料。
[Abstract]:In this study, 93 alfalfa (Medicago) Germplasm from home and abroad were collected as test materials. The genetic diversity of biological characteristics, yield, quality and molecular level and the differential expression of protein groups were analyzed in the field and indoor, and the salt tolerance of 10 Wild Alfalfa materials was evaluated in order to optimize the identification of germplasm materials. The relevant research methods of evaluation were used to evaluate the importance of related morphologic and physiological and biochemical indexes, and to select excellent germplasm materials, and to provide material and basis for the selection of new varieties of alfalfa. The experimental data were analyzed by correlation analysis, principal component analysis, path analysis, weighted subordinate function method and grey correlation analysis. Fruit: (1) among the agronomic traits, the correlation between yield and stem diameter and plant height was larger, and the correlation coefficient was 0.39 and 0.30 respectively. The correlation degree of grey correlation analysis showed that the correlation range of each character and yield was 0.72 ~ 0.84, from large to small, the number of growth rate plant number of plant height and fresh ratio plant stem and leaf ratio was the number of plant growth rate. Stem diameter; the correlation range between the tested materials and the standard varieties was 0.49 to 0.66. When the genetic distance was 7, 93 materials were gathered into 4 classes, of which 4 were the best types of materials, including M1-5 (Seth), M1-7 (West Blend), M1-54 (middle alalex No. 2), M1-86 (3). (2) 30 to 30 alfalfa varieties with 30 genotypes to SSR markers. A total of 565 alleles were detected and 18.83 alleles per locus were amplified on the average. The polymorphism information content (PIC) was 0.84, and the polymorphism information content of the selected SSR markers was divided into 2 populations by Structure analysis of more than 0.6.120 alfalfa genotypes, among which the population I included most of the alfalfa in China. There are 96 genotypes in 9 countries, and 24 genotypes of population II are derived from 6 countries. The geographical regions are Asia, Europe, America and Oceania. The genetic composition of the two populations is complex, which can be further divided into 2 and 4 subgroups, among which alfalfa is divided into subgroups and alfalfa 2 in alfalfa remains. (3) in the identification of salt tolerance, the index of salt tolerance of Alfalfa at the germination stage was the dry weight and germination rate of seedlings, which were 0.752 and 0.745 respectively. The significant correlation of salt tolerance in seedling stage was leaf water content and chlorophyll content, 0.741 and 0.660. with strong salt tolerance were M3-8 (from Inner Mongolia). The Mount Daqing flattened bean in ancient Tongliao. (4) the highest total yield material was M4-5 (new strain 1) in the new strain identification test. The results of 55.92t/hm2. correlation analysis and path analysis showed that the first straw has the greatest contribution to the total hay yield. The relative feeding value of the new strain 1 and 2 in the year of the trial was twelfth and eleventh respectively. The yield and relative feeding value of surprise and prairie 3 alfalfa were in the top 5, for high yield and quality materials. (5) the protein group analysis of alfalfa leaves with large difference in leaf type was analyzed by TMT technology, and 53 proteins were identified at the 0.05 level, the difference was greater than 1.2 times, the expression of three leaf clover leaves was up 29, down 24.G In the O annotation, 40% of the difference protein participates in the biological process, 43% participates in the molecular function and 17% participates in the cell composition. In the multileaf alfalfa, the main functions of the high differentially expressed proteins are cell differentiation, precursor metabolites and energy and photosynthesis. These physiological processes are beneficial to the generation of new tissues and promote leaf birth. In the long.KEGG pathway annotation, the first 5 pathways with more differentially protein are carbon metabolism, splice body, biosynthesis of amino acids, endocytosis, and starch and sucrose metabolism. The most expressed proteins in the multileaf alfalfa are mainly involved in the first 4 pathways, which enhance the carbon metabolism of alfalfa, promote the maturation of RNA and increase the synthesis of amino acids. It is beneficial for plants to accumulate organic compounds, promote cell metabolism and growth, and thus increase production. Finally, the raw materials are selected as recommended new varieties of Alfalfa 2, new strain 1, surprise and grassland 3 alfalfa and Tongliao Mount Daqing wild flattened bean material.
【学位授予单位】：内蒙古农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S54

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