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小麦骨干亲本及其后代田间试验设计和分析方法
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摘要
在小麦育种中骨干亲本是改良品种的基础,没有骨干亲本的优良遗传性状,优良的小麦新品种是难以育成或发展起来的。不仅我国绝大部分小麦大面积推广品种是骨干亲本的后代,而且还由其衍生出许多具有广泛应用价值的亲本育种材料。但是由于试验材料是骨干亲本及其衍生后代组成的,这些亲本和它们的衍生系(后代)具有亲缘关系。对于大数量具有亲缘相似性特点的骨干亲本及其衍生后代材料的田间试验及其相应的分析方法值得探讨研究。
     本研究以2007-2008年陕西杨凌矮秆小麦骨干亲本及其衍生后代共254份材料为研究对象,通过前期确定大数量骨干亲本及其衍生后代品系试验设计方案(具有分层结构的设计),后期利用分层混合模型来解决材料间相关性,估计品种效应,再利用多重比较、线性组间比较、育种值估计、多元统计、综合评价等分析方法,阐明不同世代的骨干亲本和后代的性状差异,深入分析骨干亲本及其衍生后代之间的性状演变趋势和亲本对后代的贡献大小。另外通过对2007-2008年陕西杨凌模拟产生的小区产量数据和2007-2008年山东泰安实测小区产量数据进行分析,筛选出相对效率较高且稳定的模型方法为模型的选择提供依据。最后以R语言环境为开发平台以R-Commander为设计模板,加载了相应的分析模块,方便了试验者和科研人员在今后针对具有相关性材料的大数量区域试验中进行分析操作。
     1.在处理设计中针对骨干亲本衍生群内的亲缘相似性特点,按照分层设计思路,遵照区组内相似性及各衍生世代相对平衡原则,进行处理分层的不完全区组设计。在环境设计中采用双向田间主区区组排列、主区区组内处理拉丁方排列及设置主区中心对照、随机对照、侧对照设计。
     2.线性固定模型和分层混合模型的14个性状处理效应估计标准误的合计值分别为187.58及120.89,分层混合模型处理估计误差比固定模型降低35.55%。可以看出采用重复内区组分层混合模型可有效的降低处理效应的估计误差,建议在大数量处理且数据缺失不平衡时使用分层混合模型进行处理效应的估计。通过ICC(组内相关系数)值,发现:其中小区产量、分蘖数、叶长、株高4个性状具有较高的ICC值说明这4个性状组内各个体间存在较高的相似性。从两种多重比较方法进行分组后可知,Tukey检验得到字母重叠的分组结果,它比较适用于样本量较少时进行深入比较和细致分析。Scott-Knott检验得到字母不重叠的分组结果,它在大样本时结果表示简洁且易于解释。因此建议在大样本时使用Scott-Knott检验的分组结果较好。
     3.本研究对3个骨干亲本阿夫、碧玛4号、南大2419及其衍生系谱品种在2008年陕西杨凌进行了12个性状鉴定。从骨干亲本与衍生子一代12个性状平均值比较来看,骨干亲本阿夫将小区产量、分蘖数、抗条锈病、穗粒数、穗粒重、叶长、叶宽等7个性状的较高基因累积水平的遗传背景传递给阿夫衍生子一代,而在抽穗期、每穗小穗数、千粒重、穗长、株高等5个性状的基因累积水平由于其他亲本影响而得到改进。
     骨干亲本碧玛4号将小区产量、分蘖数、抗条锈病、每穗小穗数、穗长、穗粒数、穗粒重、叶长、叶宽等9个性状的较高基因累计水平的遗传背景传递给碧玛4号衍生子二代,而在抽穗期、千粒重、株高等3个性状的基因累计水平由于其他亲本影响而得到改进。
     骨干亲本南大2419将小区产量、分蘖数、每穗小穗数、叶宽等4个性状的较高基因累积水平的遗传背景传递给南大2419衍生子一代,而在抽穗期、抗条锈病、千粒重、穗长、穗粒数、穗粒重、叶长、株高等8个性状的基因累积水平由于其他亲本影响而得到改进。从9个性状的分析上看出,阿夫、碧玛4号、南大2419等3个骨干亲本的衍生子一代、子二代、子三代、子四代中都保持稳定,由此说明骨干亲本都将良好遗传背景稳定地传递给衍生后代。
     从阿夫、碧玛4号、南大2419等3个骨干亲本的衍生后代12个性状育种值分析来看,阿夫亲本给后代的有利基因效应传递较大的是抗条锈病、高穗粒数、高穗粒重、长叶、宽叶等5个性状。其中有利基因效应传递较大的是抗条锈病、长叶。并且具有不利基因效应的每穗小穗数的育种值很小,由此说明阿夫亲本给后代带来不利基因的遗传负荷很小。碧玛4号亲本给后代的有利增效基因效应传递最大的是多分蘖数,但是也在低小区产量、感条锈病、低小穗数、短穗、低穗粒数、低穗粒重、短叶、窄叶、高秆等9个性状给后代带来不利基因效应的遗传负荷。南大2419亲本给后代的有利增效基因效应传递较大的是多小穗数、高千粒重、长穗、高穗粒重等4个性状,但是也在少分蘖数、感条锈病、短叶、高秆等4个性状给后代带来不利基因的遗传负荷。
     4.采用欧氏距离类平均法对17个材料进行聚类分析。分为8个不同的类群:类群Ⅰ为碧玛4号;类群Ⅱ为阿夫;类群Ⅲ为碧玛4号中间亲本;类群Ⅳ为南大2419;类群Ⅴ为阿夫子一代、阿夫子二代、阿夫子三代、阿夫子四代;类群Ⅵ为南大2419子一代、南大2419子二代、南大2419子三代、南大2419子四代、南大2419子五代;类群Ⅶ为阿夫子五代;类群Ⅷ为碧玛4号子二代、碧玛4号子三代、碧玛4号子四代。通过对应分析可揭示每个类群的特征,最大程度的体现了骨干亲本及其各世代衍生后代间的相似性和差异性。通过因子载荷分析可将八个因子命名为:穗粒数量和重量因子、粒重因子、生育期因子、小区产量因子、叶宽因子、每穗小穗数因子、株高因子、叶长因子。其中穗粒数量和重量因子由于方差贡献最大达到14%,此因子对产量的形成起着决定作用。通过改进雷达图综合评价分析得到各样本综合得分,其中前五名的世代依次是:南大2419子五代、南大2419子四代、阿夫子三代、阿夫子四代、南大2419子二代,综合得分最低的是:碧玛4号。由综合得分中三个亲本基因型值和后代估计育种值来看:南大2419亲本在综合得分上贡献给后代的有利增效基因潜力较大;碧玛4号亲本在综合得分上贡献给后代的不利减效基因潜力较大.三个亲本对下代的综合传递能力:碧玛4号最强,南大2419其次,阿夫最弱。
     5.通过对2007-08年陕西杨凌矮杆小麦100次模拟小区产量数据的原始线性模型进行有效性分析,结果表明:三个固定效应:重复、处理、重复内区组的多套模拟数据计算的F值中超过50%大于原始数据F值,认为三个固定效应的原始概率值可以接受,即原始线性模型方差分析结果是有效的,可利用模型产生的模拟数据进行分析。通过两个地点数据分析综合说明了三次重复简化广义格子设计分析(SGL)模型能更进一步控制田间试验误差,且增加重复数可以更为精确的估计处理效应,在山东实测数据实验中2次重复(SGL)模型就能达到三次重复随机完全区组分析(RB)模型的试验效率水平。进一步证明了大数量试验中重复次数为3次重复的(SGL)模型能有效提高试验效率。
     6.本研究以R语言环境为开发平台以R-Commander为设计模板,遵循软件工程理念,采用模块化设计方法,从研究原有的混合模型分析程序、调整均值估计程序、多重比较方法程序和线性分组比较程序入手,构建了以鼠标、菜单操作的软件分析模块,并初步得到以下结果:(1)构建了混合模型分析模块,利用该模块可用于试验设计中的裂区设计分析,巢式设计分析等,试验者和科研人员可方便的在参数菜单中选择最大似然估计(ML)和限制性最大似然估计(REML)并在固定效应和随机效应参数中对分类因子各水平在不同层次上进行效应估计。(2)增加了调整均值的估计分析模块,方便在以后的协方差分析中对处理因子各水平效应均值进行有效的调整以及在非均衡数据情况下对均衡情况的边缘均值做简单地估计。(3)增加了多重比较分析、线性分组比较模块,并对经典的方法做了扩增(如:加入各种P值调整的多重比较方法和针对任意对比的Scheffe、Tukey线性分组比较法)丰富了试验者和科研人员对多重比较、线性分组比较方法的选择。
Founder parent is a base of improved variety in wheat breeding.A new and good wheat variety cannot be bred or developed without the integrated elite characters of Founder parent.Not only the descendant of Founder Parent were selected wheat varieties with large area as materials in most areas of china.But also It derives a lot of breeding parents who have a widely application.But because of experimental material are made up of founder parents and their progeny. And founder parent and its derivative progeny have a relative affinity.Field trial and the corresponding analytical methods for materials of founder parent and its derivative progeny with basing on the Large quantity and Affinities
     similarity which are worth disscussing and researching. This study is consider that the 254 materials of founder parents and their progeny from Yangling dwarf wheat variety trials in 2007-2008.The scheme of design(the design of hierarchical structure) for the number of founder parent and its derivative progeny was determined in prophase.The strain effects and correlation of the material are being solved by using hierarchical mixed model,And significant differences in traits between founder parents and their progeny in different generations was illustrated using methods of Multiple Comparison,Linear Grouped Contrast,Breeding value estimates,Multivariate Statistics analysis and Comprehensive Evaluation.The evolutionary tendency and contribution between founder parents and their progeny has been analyzed more further.Through analyzing the plot yield data which would be partly simulated from YangLing in 2007-2008 of ShanXi Province,and partly measured from TaiAn in 2007-2008 of ShanDong Province.The more relative efficiency and stable models were selected by simulation.Finally,We set R language environment as Development platform,and set R-Commander as Design Template.We constructed a software analytical module.It convenient to analyze and manipulate in the correlation of the material and large number of reginal trials and in the future for experimentalist and research worker.
     1. Based on the characteristic affinities similarity in founder parent and its derivative progeny of treatment design. According to the ideas of hierarchical design.“intrablock similarity”and“Relative balance on each derivative generation”should be basic principle during conducted incomplete block design with hierarchical treatment. Using Two-direction main area block Arrange in field, Latin square Arrange in intrablock treatment of main area and Setting center CK,random CK,side CK in main area during Environmental design.
     2. The S.E.means on the estimated treatment effects for 14 traits of general linear model and hierarchical mixed model value were 187.58 and 120.89,respectively,the S.E.means on the estimated treatment effects of hierarchical mixed model value was lowered 35.55% than general linear model value. It is observed that hierarchical mixed model on grouping block within repeat could effectively reduce the errors on the estimated treatment effects.Suggestions are useing hierarchical mixed model to estimated treatment effects when it has a Large number of treatments and Data are missing and imbalancing.Through ICC(Interclass Correlation Coefficient),We find that Four traits:plot yield,tillernumber,leaf length,plant height,which has higher ICC values.It shows that individual have higher Similarity within class in 4 traits. according to grouping results by two methods on multiple comparisons.the letter was overlap in Tukey grouping test. It is delicate, Intensive to be Compared on small samplesize.the letter wasn’t overlap in Scott-Knott grouping test.It is succinct,easy to be explained on large samplesize. It is thus suggested that Using the scott-knott method can conduct better the grouping results on large samplesize.
     3. Base on 3 founder parents:Funo,Bima4,Nanda2419 and its derivative progeny as research object.then their 12 traits were identified in YangLing of ShanXi province in 2008. In view of the means for 12 traits from founder parent and its first filial generations. Founder parent of Funo can transfer genetic background which have high cumulative level on alleles from 7 traits:plot yield, tillernumber, the ability of stripe rust-resistant, grain number per spike, grain weight per spike, leaf length, leaf width to Its first filial generations. But the cumulative level on alleles from 5 traits: heading stage, spikelet number per spike, 1000 grains weight, ear length, plant height are improved due to the influence of Other parents.
     Founder parent of Bima4 can transfer genetic background which have high cumulative level on alleles from 9 traits:plot yield, tillernumber, the ability of stripe rust-resistant, spikelet number per spike, ear length,grain number per spike, grain weight per spike, leaf length, leaf width to Its second filial generations.But the cumulative level on alleles from 3 traits: heading stage, 1000 grains weight, plant height are improved due to the influence of Other parents. Founder parent of Nanda2419 can transfer genetic background which have high cumulative level on alleles from 4 traits:plot yield, tillernumber, spikelet number per spike, leaf width to Its first filial generations.But the cumulative level on alleles from 8 traits: heading stage, the ability of stripe rust-resistant, ear length,grain number per spike, grain weight per spike, leaf length,plant height are improved due to the influence of Other parents. By analysing the 9 traits .It has remained basically steady on 3 founder parents:Funo,Bima4,Nanda2419 and their first filial generations,second filial generations,third filial generations, fourth filial generations, fifth filial generations, It shows that the Founder parent can Steady transfer good genetic background to its derivative progeny.
     In view of the breeding value for 12 traits from 3 founder parent:Funo,Bima4,Nanda2419 and its derivative progeny.Founder parent of Funo can transfer favorable genetic effect from 5 traits: the ability of stripe rust-resistant, high grain number per spike, high grain weight per spike, leaf length, leaf width to Its derivative progeny.And stripe rust-resistant, leaf length can transfer more favorable genetic effect to Its derivative progeny.breeding value which have unfavourable genetic effect is small on spikelet number per spike.It shows that Founder parent of Funo can transfer smaller genetic load which have unfavourable genetic effect to Its second filial generations. Founder parent of Bima4 can transfer most favorable and additive genetic effect from 1 traits: tillernumber to Its derivative progeny.But Founder parent of Bima4 can transfer genetic load which have unfavourable genetic effect to Its derivative progeny from 9 traits: low plot yield, Feeling stripe rust, less spikelet number per spike,short ear length,less grain number per spike, light grain weight per spike,short leaf, narrow leaf,high plant height.Founder parent of Nanda2419 can transfer most favorable and beneficiated genetic effect from 4 traits: more spikelet number per spike,high 1000 grains weight,long ear length, heavy grain weight per spike to Its derivative progeny.But Founder parent of Nanda2419 can transfer genetic load which have unfavourable genetic effect to Its derivative progeny from 4 traits: less tillernumber, Feeling stripe rust, short leaf, high plant height.
     4. The 17 grouping materials were clustered by using Euclidean distance and group-average method.It suggested that the materials could be divided into 8 groups: GroupⅠis Bima4.GroupⅡis Funo.GroupⅢis inter parents of Bima4.GroupⅣis Nanda2419. GroupⅤare first filial generations, second filial generations, third filial generations, fourth filial generations, fifth filial generations of Funo. GroupⅥare first filial generations, second filial generations, third filial generations, fourth filial generations, fifth filial generations of Nanda2419. GroupⅦis fifth filial generations of Funo. GroupⅧare second filial generations, third filial generations, fourth filial generations of Bima4 respectively.through a Correspondence Analysis, announcing to the characteristic of each groups: The results maximum present Similarity and dissimilarity between Founder parents and its derivative progeny.Through the factor loading was estimated by principal component analysis. The 8 factors will be named:The factor of grain number and weight per spike; The factor of 1000 grains weight; The factor of growth period ; The factor of plot yield; The factor of leaf Width; The factor of spikelet number per spike; The factor of plant height; The factor of leaf length;We find that the factor of grain number per spike and grain weight whose contribution of variance accounted to 14%.This factor play a decisive role in yield-building.Through the comprehensive evaluation and analysis by improved Radar chart.We can get the comprehensive score for each sample. The top 5 generations followed by:the fifth filial generations of Nanda2419,the fourth filial generations of Nanda2419,the third filial generations of Funo,the fourth filial generations of Funo,the second filial generations of Nanda2419. The lowest comprehensive score is Bima4. In view of the Comprehensive scores for genotype value and EBVs from3 founder parent and its derivative progeny.Founder parent of Nanda2419 can transfer more favorable and beneficiated genetic effect to Its derivative progeny. Founder parent of Bima4 can transfer more unfavorable and reductive genetic effect to Its derivative progeny.Comprehensive transmission ability for 3 founder parents and its derivative progeny:The best is Bima4,The next is Nanda2419,The weakest is Funo.
     5. Through the effectiveness analysis of the Primitive linear model which based on the 100 times of simulation data for plot yield from Yangling dwarf wheat variety trials in 2007-2008.The results show that Over 50 percent of F value which was calculated by the 100 times of simulation data for Three fixed effects: Repetition, Treatment,Block within Repetition is greater than F value which was calculated by raw data. We think original probability value from three fixed effects is quite acceptable. It is show that the result of ANOVA is effectiveIt is available to produced simulation data by the Primitive linear model, and It can be analyzed.
     Through analysis the data from 2 locis, it has been confirmed that the simplified generalized grid design analysis (SGL) is better control the error from field experiment,and It Can be more precise estimateeffects of treatments by increasing the number of Repetition.Test efficiency level in (SGL)model on 2 replications as well as Random complete block designs(RB)model on 3 replications. It is proved that (SGL)model on 3 replications can highly increase test efficiency in large number of experiment.
     6. This research based on R language environment platform.And taken R-Commander as the design of the template,established a module of the mixed models.Thismethod could be used in split plot design analysis and Nested design analysis in Design of Experiment.Experimentalist and Research worker can easily choose the maximum likelihood estimation(MLE)and the restricted estimation maximum likelihood (REML) in the parameter menu.And It can estimated the effects for each levels in grouping factors of different hierarchies in Fixed effects and random effects parameters.an increase of estimating adjustment mean modules.It can easily adjust means in each levels of treatment factors for Analysis of covariance in the future,and Marginal Means were simply estimated for assuming the balanced situation in unbalanced data.an increase of Multiple comparison analysis and Linear grouping contrasts modules,and It has increased some methods on the previous(for example:Added some methods of Multiple comparison analysis for P value adjustment and Scheffe、Tukey Linear grouping contrasts for arbitrary contrasts).It enrich the methods of Multiple comparison analysis and Linear grouping contrasts which were selected by Experimentalist and Research worker.
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