1、 硕士毕业论文硕 士 学 位 论 文多花黑麦草杂交、回交群体的遗传变异及生产性能评价学科专业：草业科学硕士毕业论文I摘 要本研究采用 SSR 分子标记及表型性状分析的方法，对多花黑麦草(Lolium multiflorum)杂交及回交群体进行遗传分析，旨在对多花黑麦草杂交回交群体的改良效果进行评价。另外研究还通过对 14 个多花黑麦草品种（系）两年品比的试验，在生产性能、农艺和表型性状等方面综合评价14 个多花黑麦草品种（系）在雅安地区的适应性。主要研究结果如下：(1)利用SSR分子标记对2个多花黑麦草杂交组合5轮改良群体进行了遗传变异分析。 20对SSR引物在12个育种群体中共扩增出217个
2、条带，多态性条带比率为92.2%。两个杂交群体的不同选择世代中，分别有72.3%和75.2%的遗传变异存在于群体内，27.7%和24.8%的遗传变异存在于各群体间，群体内的遗传变异大于群体间的遗传变异。从带型分析来看，杂交改良群体中双亲共有的条带比例增多，同时缺失双亲特异性条带的比例减少。随着混合选择世代的增加，多态性条带百分率、Shannon遗传多样性指数及Neis基因多态性都呈递减趋势，表明群体内的遗传差异随着选择世代的增加呈下降趋势。研究结果将为多花黑麦草杂交选择育种提供基本信息。(2) 对 2 个多花黑麦草杂交组合材料 5 轮改良群体进行了表型性状分析。通过变异系数和方差分析等统计方法
3、表明，供试的 2 个杂交组合内形态性性状上的遗传差异显著，且随着混合选择世代的增加各表型性状的变异系数相对减小。且 2 个杂交组合各世代间在表型性状上的显著性差异随世代的增加而增加，世代内的显著性差异随世代的增加而减少。（3）采用回交对长江2号多花黑麦草的茎粗等主要表型性状进行改良，效果明显。14份材料的表型性状分析结果表明：杂交组合C2、C7及C8的茎粗及节间长表现优于亲本长江2号。经过一轮回交，各回交组合除含有目标性状外，其他性状也发生了不同程度的回复。对各改良材料群体进行SSR分析表明， 20对SSR 引物在7个育种群体中共扩增出 217个条带，多态性条带比率为92.2%。 SSR分子标
4、记水平上的遗传相似性分析表明，回交组合表型性状上的变化原因是发生在遗传物质变化上的。这为长江2号多花黑麦草今后的进一步改良提供基础信息。（4）对创制的新材料，以国审品种为对照，进行品种比较试验。结果表明：新品系YA03和YA08的冬季生长速度相对较快，与剑宝、阿伯德差异显著（p0.05），具有明显的冬季生长优势。产草量方面，新品系YA17表现最好，长江2号次之。而新品系YA03前三茬产量优势明显，20112013两年度平均干草产量可达12994kg/hm 2，比对照增产11.5015.66%；两年度平均鲜草产量为98973kg/hm 2，比对照增产9.4712.82%。茎叶比方面，阿伯德叶片含
5、量最高（茎叶比为1：1.86），其次为新品系YA03（茎叶比为1：1.68）。而在物候期、营养成分等方面，各多花黑麦草品种（系）差异不显著。综合得知，新品系YA17和YA03具有产量较高，叶片含量丰富，较适合川西南区冬闲田稻草轮作。关键词：多花黑麦草；杂交育种；回交育种；混合选择；表型性状；SSR；生产性能。硕士毕业论文IIAbstractIn this study, SSR markers and agronomic traits were used to evaluate the genetic diversity in the crossbreeding groups and backc
6、ross breeding groups of annual ryegrass. As well, According to comparison of 14 lolium mulitiflorum cutivars and strains on production perfomance, growing speed, agronomic characteristics in two years, adaptability of these lolium multiflorum was assessmented in Yaan city.The results were as followe
7、d:（1）SSR markers were used to evaluate the genetic diversity in two crossbreeding groups of annual ryegrass after 5 cycles mass selection. With the 20 primer pairs selected, a total of 217 bands were detected in the all populations, the percentage of polymorphic bands was 92.2%. In the two crossbree
8、ding groups, 72.3% or 75.2% of the genetic diversities were estimated intra-populations and 27.7% or 24.8% were estimated inter-populations. The inter-populations genetic diversity was much more than the intra-populations. After the bands analysis, the amplified results showed that the crossbreeding
9、 groups had higher mutual bands of parents with progenies but lower non parental specific bands. With the proceeding of the mass selection in crossbreeding groups, the percentage of polymorphic bands in populations declined as well as the Shannon index and Neis gene diversity. It is showed that gene
10、tic diversity of per population declined under the mass selection. The results of this study will provide some basic information to the ryegrass crossbreeding.（2）Seven charateristics of two Lolium multiflorum hybrid combinations were evaluated at heading stage and flowering stage. And statistical an
11、alysis such as coeffcient of variation and variance analysis indicated that there was singificant difference in two combinations. With the proceeding of the mass selection in crossbreeding groups, the variation varied declined, the significant difference among the crossbreeding groups increased, but
12、 the significant difference inner the crossbreeding groupsdeclined.（3）Backcross method was used to improving the annual ryegrass Changjiang No.2.The analysis of the agronomic traits of the 14 tested materials showed that the significant heterobeltiosis were found in the stem diameter and the interno
13、des length of the hybridized combination C2, C7 and C8. After the backcross, the agronomic traits of each backcross combination had recovered at different degrees. The genetic analysis by SSR markers showed that a total of 217 bands were detected in the all populations, with the 20 primer pairs sele
14、cted, the percentage of polymorphic bands was 92.2%. The genetic similarity analysis by SSR markers showed that the variation of agronomic traits in backcross 硕士毕业论文IIIwas caused by the changes occurred in the genetic materials. The results of this study will provide some basic information to the ry
15、egrass backcross breeding.(4) The production performances of 14 varieties of annual ryegrass in Yaan, southwest of Sichuan province, were studied in present study. The results showed that the growth rates of the new lines YA03 and YA08 were faster than Jumbo and Aubade (p0.05). During the two years
16、from 2011 to 2013, the yields of new line YA17 were the highest, and YA03 also had advantagesin the yields of the first three crops.The fresh yield of YA03 achieved 12994kg/hm2 on an average, increased with the range from 11.50% to 15.66%,comparing with Changjiang No.2, Tetragold andAubade(p0.05). T
17、he hay yield also increased with the range from 9.47% to 12.82%, achieved 98973 kg/hm2. In addition, Aubade and YA03 were very rich in leaves. Meanwhile, there were no significant differences of the phenologyand the main nutrient content among the 9 tested varieties. Thus, the new lines YA17 and YA0
18、3 with higher yields and rich leaves were more suitable for planting in southwest of Sichuan province.Key words:Lolium multiflorum; Crossbreeding; Backcrossbreeding；Mass Selection；Agronomic traits；SSR; Production Performance.硕士毕业论文IV目录第一章 前言 .11 研究的目的及意义 .12 国内外的研究进展 .12.1 国外多花黑麦草的新品种选育研究 .12.2.2 选择
19、育种 .32.2.3 杂交育种 .32.2.4 诱变育种 .42.2.5 倍性育种 .42.3 分子标记在黑麦草种质资源研究中的进展 .43 研究内容与技术路线 .53.1 多花黑麦草杂交及回交群体的 SSR 多样性变化 .53.2 多花黑麦草杂交及回交群体的表型性状变化 .53.3 14 个多花黑麦草品种（系）在川西南地区生产性能综合评价 .63.4 技术路线图 .6第二章 连续混合选择下多花黑麦草杂交群体的 SSR 多样性变化 .71 材料与方法 .71.1 试验材料 .71.2 试验方法 .81.2.1 DNA 提取及检测 .81.2.2 SSR-PCR 反应及电泳检测 .81.2.3
20、数据统计及分析 .82 结果与分析 .92.1 SSR 引物的筛选及其多态性分析 .92.2 各杂交后代群体的 SSR 扩增条带分析 .93 讨论与小结 .12第三章 连续混合选择对多花黑麦草杂交群体表型性状的影响 .141 材料与方法 .141.1 试验地简介 .141.2 试验材料 .141.3 田间管理 .151.4 试验内容与方法 .15硕士毕业论文V1.5 数据处理 .152 结果与分析 .152.1 形态学指标 .152.2 方差分析 .173 讨论与小结 .18第四章 长江 2 号多花黑麦草主要表型性状的回交改良分析 .201 材料与方法 .201.1 试验材料 .201.2 试
21、验方法 .201.3 DNA 提取及检测 .211.4 SSR-PCR 反应及电泳检测 .211.5 数据统计及分析 .211.5.1 表型性状数据分析 .211.5.2 SSR 分子标记结果统计及分析 .222 结果与分析 .222.1 具有抗倒伏相关性状的多花黑麦草杂交子代的筛选 .222.2 回交子代的回交效果分析 .222.2.1 回交子代表型性状水平上的回交效果分析 .222.2.2 SSR 分子标记水平上的回交效果分析. .233 讨论与小结 .24第五章 几个多花黑麦草品种（系）在川西南地区生产性能综合评价 .261 材料与方法 .261.1 试验时间及试验地概况 .261.2 试验材料及来源 .271.3 试验方法与测定项目 .271.3.1 小区设计 .271.3.2 观测项目及方法 .271.4 田间管理 .282 结果与分析 .282.1 生育期 .282.2 植株生长速度 .292.3 主要营养成分 .292.4 表型性状分析 .302.5 产草量分析 .31硕士毕业论文VI2.6 茎叶比 .343 讨论与小结 .
