MAGIC群体的遗传特征及其在作物耐逆研究上的应用

多亲本高世代互交（multi-parent advanced generation intercross,MAGIC）群体是近年来发展起来的新一代遗传作图及育种群体。MAGIC群体最初是以研究动物及人类复杂性状遗传基础为目标而构建的基于多亲本的重组近交群体,随后将其构建方法衍生到植物中应用。MAGIC群体应用于作物遗传育种,可以建立包含无限多株系的群体,主要的优势是拥有大量可利用的多样性遗传基因池。亲本可选用育种中性状优异的材料,通过多次重组创造大量的遗传变异。群体中选出的优良株系可用作育种中间材料或直接组配新品种,也可灵活应用于数量性状位点（quantitative trait locus,QTL）的精确遗传定位分析,真正做到育种群体和定位群体的有机整合。作物耐逆性大多是由QTL控制,因此,主要围绕MAGIC群体的定义、构建流程、遗传特征及其在作物耐逆性研究上的应用和发展前景作综合性阐述。     

Comparison of quantitative trait loci for 1,000-grain weight and spikelets per panicle across three connected rice populations

The ability to detect quantitative trait loci (QTLs) in a bi-allelic population is often limited. The power of QTL detection and identification of the most beneficial allele at each QTL could be greatly improved by comparing QTLs among different populations derived from connecting multi-parents. In this study, three sets of connected recombinant inbred lines (RILs) derived from the crosses between Zhenshan 97 and Minghui 63 (PZM), Zhenshan 97 and Teqing (PZT), and Minghui 63 and Teqing (PMT), respectively, were used. QTL analyses for the number of spikelets per panicle (SPP) and 1,000-grain weight (TGW) were performed in PZT, and five SPP QTLs on chromosomes 1, 6, and 7 and two TGW QTLs on chromosome 1 were detected. QTL for SPP was also identified in PMT, and six QTLs were detected on chromosomes 1, 2, 3, 6, and 7 in this population. In an earlier study, we identified five SPP QTLs and four TGW QTLs in PMT and nine TGW QTLs in PZM. Comparison of the QTL mapping results of these two studies showed that one QTL was common to the three populations, 11 QTLs were detected in two populations, and six QTLs were found in only one population. Comparison of genetic effect and the action direction of the QTLs detected in the three populations showed that additive effects of QTLs estimated in different populations were also expressed additively among three parental alleles. Additive effects of SPP7a estimated in three near-isogenic line F₂ populations supported this finding. Based on these results, we suggest that pyramiding the most beneficial alleles among the three parents could efficiently improve rice yield.     

Identifying quantitative trait loci for the general combining ability of yield-relevant traits in maize

Maize is the most important staple crop worldwide. Many of its agronomic traits present with a high level of heterosis. Combining ability was proposed to exploit the rule of heterosis, and general combining ability (GCA) is a crucial measure of parental performance. In this study, a recombinant inbred line population was used to construct testcross populations by crossing with four testers based on North Carolina design II. Six yield-relevant traits were investigated as phenotypic data. GCA effects were estimated for three scenarios based on the heterotic group and the number of tester lines. These estimates were then used to identify quantitative trait loci (QTL) and dissect genetic basis of GCA. A higher heritability of GCA was obtained for each trait. Thus, testing in early generation of breeding may effectively select candidate lines with relatively superior GCA performance. The GCA QTL detected in each scenario was slightly different according to the linkage mapping. Most of the GCA-relevant loci were simultaneously detected in all three datasets. Therefore, the genetic basis of GCA was nearly constant although discrepant inbred lines were appointed as testers. In addition, favorable alleles corresponding to GCA could be pyramided via marker-assisted selection and made available for maize hybrid breeding.     

Quantitative trait loci mapping of forage agronomic traits in six mapping populations derived from European elite maize germplasm

Four agronomic traits were analysed including dry matter concentration (DMC) and dry matter yield (DMY) for stover, plant height (PHT) and days from planting to silking (DPS). We mapped quantitative trait loci (QTL) in three populations with doubled haploid lines (DHL), one RIL population and two testcross (TC) populations derived from crosses between two of the four populations mentioned above to elite tester lines, based on field phenotyping at multiple locations and years for each; 146–168 SSRs were used for genotyping of the four mapping populations. Significant high phenotypic and genotypic correlations were found for all traits at two locations, while DMC was negatively correlated with the other traits. A total of 42, 41, 54, and 45 QTL were identified for DMC, DMY, PHT, and DPS, respectively, with 9, 7, 12, and 7 major QTL for each trait. Most detected QTL displayed significant interactions with environment. Major QTL detected in more than two populations will contribute to marker‐assisted breeding and also to fine mapping candidate genes associated with maize agronomic traits.     

利用品质性状的回交选择导入系挖掘水稻抗纹枯病QTL

将优质、抗纹枯病的高秆供体Tarom Molaii和Binam导入半矮秆IR64和特青背景,培育品质性状回交选择构建的4个导入系群体IR64/Tarom Molaii、特青/Tarom Molaii、IR64/Binam和特青/Binam,定位了影响水稻抗纹枯病病级(disease scale, DS)、相对病斑高度(relative lesion height, RH)和株高(plant height, PH)的QTL。结果表明,4个导入系群体的DS与RH高度相关,两者与PH呈显著负相关。导入系后代各性状均呈现超亲分离,出现抗性明显优于双亲的抗病个体,其中40%左右属半矮秆抗病类型。采用单向方差分析,在这4个群体中分别定位到10、8、8和6个影响3个性状的QTL,多数基因座上降低DS和RH即增强抗病性同时增加株高的等位基因均来自两个供体。未在同一供体两个不同背景下检测到影响3个性状的相同QTL,表明抗纹枯病QTL表达有明显的遗传背景效应。PH与DS及PH与RH被定位在同一个显著标记位点的QTL数分别占两个性状QTL总数的38%和52%,表明水稻纹枯病抗性与株高关系密切,两者存在许多连锁位点。与以往相同群体品质性状QTL的定位结果相比,发现品质性状QTL与抗纹枯病QTL大多分布在染色体的不同区域,彼此独立遗传。对利用目标性状选择导入系定位非目标性状QTL的效果、影响因素及育种应用进行了探讨,强调了目标性状选择导入系对非目标性状QTL发掘及育种应用的重要性。     

陆地棉光合相关性状的QTL定位分析

光合作用是棉花产量的主要物质来源。本研究以高光效陆地棉冀优861和低光效陆地棉新陆早25号为亲本组配的196个F₂单株为作图群体,利用SSR（Simple Sequence Repeat）标记构建陆陆杂交遗传连锁图谱,共有30个标记位点连锁,包含4个连锁群,全长244.4cM。利用QTL IciMapping 4.1软件的完备区间作图法对冀优861×新陆早25号F_2:3家系的光合相关性状进行QTL作图分析,共定位到光合相关5个性状的10个QTLs,其中1个光合速率QTL和1个胞间CO₂浓度QTL分别定位在D3和D7染色体上。本研究为棉花光合相关性状QTL的精细定位及分离克隆打下基础,为聚合棉花高光效分子标记辅助育种提供理论依据。     

Marker-assisted selection efficiency in multiple connected populations: a simulation study based on the results of a QTL detection experiment in maize

Many studies already investigated marker-assisted selection (MAS) efficiency but mainly in biparental populations. Connected multiparental populations address a broader diversity and confer a gain of power for QTL detection which must be beneficial for MAS. Our objective was to compare multiparental connected designs to biparental populations taken separately for MAS and phenotypic selection. We first detected QTL for flowering time and grain yield in an experimental maize design involving four parental inbred lines crossed to produce six different biparental populations and confirmed the advantage of multiparental connected designs over biparental populations for QTL detection. Based on these results we performed stochastic simulations to evaluate the expected efficiency of four generations of MAS and phenotypic selection. Different parameters were considered: trait heritability, genetic architecture and whether QTL were assumed to be known or have to be detected. Genetic gains were higher in the multiparental design than on average over the biparental populations considered separately, especially when favourable alleles were equally distributed among parental lines. When QTL detection was included in the simulation process, we found that type I error risk considered for declaring QTL as significant should be adapted to the design. Type I error risks leading to the best response were higher for the biparental populations than for the multiparental design. Besides addressing a broader diversity, multiparental designs increase the power of QTL detection, which reinforces their superiority over biparental designs for MAS. Application of MAS to multiparental designs therefore appears promising to accelerate genetic gain in plant breeding programs.     

数量性状基因定位研究中若干常见问题的分析与解答

QTL作图是基因精细定位、克隆以及有效开展分子育种的基础,在利用QTL作图开展数量性状基因定位研究的过程中经常会碰到一些问题,与统计方法有关的一些问题包括LOD的统计学意义是什么？检测QTL的可信度和LOD临界值的关系是什么？如何评价不同的QTL作图方法？提高QTL检测效率的途径有哪些？与遗传参数估计有关的一些问题包括QTL的贡献率是如何计算出来的？如何确定QTL有利等位基因的来源？选择基因型分析的有效性如何？复合性状是否适宜于QTL作图？与作图群体及遗传图谱有关的一些问题包括QTL作图群体中表型数据是否要求服从正态分布？加密标记是否可以显著提高QTL检测功效？缺失分子标记对QTL作图有什么影响？奇异分离标记对QTL作图有什么影响？文章试图结合笔者多年研究工作对这12个有共性的常见问题做出分析和解答,以供科研工作者参考。     

玉米衔接式单片段导入系群体的构建和评价

以生产上广泛种植的玉米杂交种豫玉22的亲本自交系87-1和综3为受体亲本,以糯质、抗病性较好的玉米自交系衡白522为供体亲本,采用回交和自交的方法,结合SSR分子标记辅助选择,分别构建了以87-1和综3为背景的衔接式玉米单片段导入系群体,并对其遗传背景、导入片段大小、数目和覆盖率等进行了评价。结果表明,87-1背景的导入系群体导入了40个供体片段,片段长度介于0.03~342.8 cM,平均长度为91.1 cM,导入片段总长为3 643.9 cM,覆盖率为48.9%;综3背景的导入系群体导入了78个供体片段,片段长度介于0.03~343.4 cM,平均长度为75.5 cM,导入片段总长为5 895.2 cM,覆盖率为79.2%。     

Identifying QTL for fiber quality traits with three upland cotton (Gossypium hirsutum L.) populations

Cotton fiber quality was quantitative trait, controlled by multiple genes. Identification of stable quantitative trait loci (QTL) effectively contributing to favorable fiber quality traits would provide the key basis for marker-assisted selection used in molecular breeding projects. Three upland cotton F₂ populations were established with a common parent Chinese cultivar Yumian 1 and three American commercial cultivars/lines (Acala Maxxa, CA3084 and TAM94L-25), each of which had unique fiber quality characteristic that was favorable economically. Three whole genome genetic maps were constructed with 323, 302 and 262 SSR loci for population (Yumian 1 × Acala Maxxa), (Yumian 1 × CA3084), and (Yumian 1 × TAM 94L-25) respectively, spanning 1,617.2, 1,639.9 and 1,441.4 cM. Based on these genetic maps and three generation phenotypic data of fiber quality traits (F₂, F_2:3 and F_2:4), 77 QTL were detected, including 19 for fiber length, 14 for fiber uniformity, 17 for micronaire, 10 for fiber elongation, and 17 for fiber strength. Among these QTL, 46 QTL were significant QTL and 31 were putative QTL, including that one QTL (qFL05.1) and four QTL (qFL23.1, qFM06.1, qFM06.2 and qFE25.1) were detected across three and two populations respectively; two QTL qFL10.1 (Yumian 1 × TAM 94L-25) and qFL15.1 (Yumian 1 × Acala Maxxa) were detected in three generations; qFM23.1, qFE18.1 and qFS21.2 detected in population (Yumian 1 × CA3084), qFE10.1, and qFS10.2 detected in population (Yumian 1 × TAM 94L-25), and qFS15.1 detected in population (Yumian 1 × Acala Maxxa), were all detected in two generations. Alleles underlying these stable QTL were valuable candidate gene for fine mapping, cloning, and favorable gene pyramiding projects. Our study also verified that QTL mapping of fiber quality traits using multiple populations with a common parent had higher efficiency compared to single population crossed with two parents and favorable alleles contributed to QTL effect could be conferred by parents with inferior fiber quality traits.     

Multiple‐line cross QTL mapping for grain yield and thousand kernel weight in triticale

Grain yield and its component trait thousand kernel weight are important traits in triticale breeding programmes. Here, we used a large mapping population of 647 doubled haploid lines derived from four families to dissect the genetic architecture underlying grain yield and thousand kernel weight by multiple‐line cross QTL mapping. We identified 3 QTL for grain yield and 13 for thousand kernel weight which cross‐validated explained 5.2% and 48.2% of the genotypic variance, respectively. Both traits showed a positive phenotypic correlation, and we found two QTL overlapping between them. Full two‐dimensional epistasis scans revealed epistatic QTL for both traits, suggesting that epistatic interactions contribute to their genetic architecture. Based on QTL identified in our results, we conclude that the potential for marker‐assisted selection is limited for grain yield but more promising for thousand kernel weight.     

超级杂交稻‘Y两优2号’耐冷性QTL定位与杂种优势分析

挖掘耐冷基因并提高耐冷性对于保证水稻在气候变化条件下的高产稳产具有至关重要的意义。本研究利用‘远恢2号’和‘Y58S’杂交而成的超级杂交稻‘Y两优2号’的高世代重组自交系(RIL F14)276个家系作为作图群体,以SNP为分子标记构建了高密度遗传图谱,对水稻的芽期耐冷性(cold tolerance at the bud bursting period,CTBP)性状进行数量性状位点(quantitative trait locus,QTL)定位分析;同时对全世界范围内收集的水稻自然变异微核心(Minicore)种质群体进行芽期耐冷性全基因组关联分析(genome-wide association study,GWAS)。结果表明,水稻芽期耐冷性在水稻群体内呈连续分布,是由多基因控制的数量性状。同时,在RIL群体的第9号染色体上定位到1个与耐冷性性状相关的QTL,位于区间Block73479和Block72824之间,对表型变异的解释率为9.65%。进一步分析表明该QTL对水稻芽期耐冷性为负显性。结果还显示‘Y两优2号’耐冷性显著强于亲本,具有杂种优势。     

利用籼粳交探讨水稻株高和抽穗期的遗传基础

广亲和基因的发现,克服了籼粳交杂种不育的矛盾,但杂种后代株高过高、抽穗期延迟等问题仍然限制着籼粳亚种间杂种优势的利用。本试验利用一个籼粳交(珍汕97/武育粳2号)双单倍体群体(doublehaploidpopulation,DH系)及其分别与两亲本回交构建的两个回交群体,考查了株高和抽穗期的遗传,将三个群体的表型值和两个回交群体的中亲优势值进行了数量性状位点(quantitativetraitlocus,QTL)检测及效应分析,并对结果进行了比较。2个性状在3个相关群体中一共检测到了21个主效QTL和10个上位性QTL,主效QTL的数目和贡献率都比上位性QTL大,而且,在10个上位性QTL中,发生在2个主效QTL间的互作有3个,主效QTL与背景位点间的互作有6个,2个互补位点间的互作仅有1个,进一步说明了主效QTL在株高和抽穗期遗传中的重要作用。比较加性和非加性QTL的作用,发现加性效应、显性效应和上位性效应是籼粳亚种间杂种株高和生育期变化的共同遗传基础。     

Genetic analysis of plant height using two immortalized populations of “CRI12 × J8891” in Gossypium hirsutum L.

Plant height is an important plant architecture trait that determines the canopy structure, photosynthetic capacity and lodging resistance of upland cotton populations. To understand the genetic basis of plant height for marker-assisted breeding, quantitative trait loci (QTL) analysis was conducted based on the genetic map of recombinant inbred lines (RILs) derived from the cross “CRI12 × J8891” (Gossypium hirsutum L.). Three methods, including composite interval mapping, multiple interval mapping and multi-marker joint analysis, were used to detect QTL across multiple environments in the RILs and in the immortalized F₂ population developed through intermating between RILs. A total of 19 QTL with genetic main effects and/or genetic × environment interaction effects were identified on 15 chromosomes or linkage groups, each explaining 5.8–14.3 % of the phenotypic variation. Five digenic epistatic QTL pairs, mainly involving additive × additive and/or dominance × dominance, were detected in different environments. Seven out of eight interacting loci were main-effect QTL, suggesting that these loci act as major genes as well as modifying genes in the expression of plant height. The results demonstrate that additive effects, dominance and epistasis are all important for the genetic constitution of plant height, with additive effects playing a more important role in reducing plant height. QTL showing stability across environments that were repeatedly detected by different methods can be used in marker-assisted breeding.     

四向杂交设计QTL分析的极大似然方法

四向杂交（four-way cross）设计是指4个纯系亲本参与杂交衍生分离群体的一种交配设计。尽管国际上已经提出基于四向杂交设计的迭代重新加权最小平方（iteratively reweighed least squares，IRWLS）QTL作图方法，但该方法忽略了双侧标记基因型内QTL基因型的混合分布特性，当QTL位置和标记的位置不重合时作图精度较低。本文根据四向杂交设计的数量遗传模型，发展出基于四向杂交设计和混合分布理论的QTL作图的极大似然估计方法。首先利用染色体上所有标记基因型联合计算该染色体上任一假定位置QTL的条件概率，然后根据混合分布理论建立基于EM算法实现的QTL作图的极大似然估计方法。以计算机模拟数据研究了QTL遗传力、样本容量和分子标记信息含量3个因素对方法的影响，结果表明,（1）在QTL的被发现能力上，标记信息不完全的四向杂交设计仅略低于信息完全时的四向杂交设计；（2）随着QTL遗传力、样本容量和标记信息含量的增大，QTL位置以及效应估计值的准确度和精确度逐步提高。     

利用相同来源F2:3和BC2S1群体定位玉米生育期QTL

以普通玉米自交系丹232和爆裂玉米自交系N04为亲本构建259个F2:3和220个BC2S1家系群体,利用SSR标记构建分子标记遗传图谱,利用复合区间作图方法对4个生育期性状进行QTL定位和效应分析。利用F2:3群体共检测到4个抽雄期QTL、6个吐丝期QTL和3个散粉期QTL。单个QTL可解释的表型变异为6.7%~18.4%,可解释的表型总变异为28.9%~50.3%,11个QTL的增效基因来自生育期较长的亲本丹232,其余2个QTL的增效基因来自生育期较短的亲本N04;BC2S1群体检测到8个与4个生育期性状相关的QTL,单个QTL可解释的表型变异为4.5%~11.6%,可解释的表型总变异为13.2%~18.5%,增效基因来自两个亲本的QTL为3个和5个。两类群体检测出QTL的数目、位置、效应和贡献率均存在较大差异,主要原因在于BC2S1群体抽样选择所引起的群体结构差异,F2:3群体显示出较高的QTL检测能力,但回交育种过程中应慎重依据F2:3群体QTL定位结果进行标记辅助选择(MAS)。     

Molecular and comparative mapping for heading date and plant height in oat

Selection of oat genotypes combining earliness and short plant height could stimulate oat cultivation worldwide. However, the mechanisms involved with the genetic control of heading date and plant height traits are not fully understood to date. This study aimed to identify genomic regions controlling heading date and plant height in two hulled by naked oat populations and to compare these genomic regions with that of other grass species. Recombinant inbred lines of each population and their parents were genotyped by a 6 K BeadChip Illumina Infinium array and assessed for heading date and plant height in two sowing dates. The quantitative trait loci (QTL) affecting these traits were detected by simple interval mapping. The two oat populations showed different genetic mechanisms controlling heading date. A major QTL was identified in one of the populations, mapped into the ‘Mrg33’ consensus linkage group from the current oat map. Two other QTL were detected into the ‘Mrg02’ and ‘Mrg24’ groups, in the second population. On the other hand, both populations presented the same genomic region controlling plant height. Six SNP markers, mapping on the same linkage group within each population, were associated with the trait, regardless the sowing date, explaining more than 20% of the phenotypic variation. Five of these six markers were mapped into three different linkage groups on the oat consensus map. Genomic regions associated with heading date and plant height in oat seem to be conserved in Oryza sativa L. and Brachypodium distachyon. Our results provide valuable information for marker-assisted selection in oats, allowing selection for earliness and plant height on early segregating generations.     

Analysis of QTL for seed oil content in <Emphasis Type="Italic">Brassica napus</Emphasis> by association mapping and QTL mapping

Increasing seed oil content is one of the most important breeding targets for rapeseed (Brassica napus). In this study, we combined quantitative trait loci (QTL) mapping and marker-trait association analysis to dissect the genetic basis of seed oil content in rapeseed. A doubled haploid (DH) population with 261 lines was grown in two highly contrasting macro-environments, Germany with winter ecotype environment and China with semi-winter ecotype environment, to explore the effect of environment effect of on seed oil content. Notable macro-environment effect was found for seed oil content. 19 QTL for seed oil content were identified across the two macro-environments. For association analysis, a total of 142 rapeseed breeding lines with diverse oil contents were grow in China macro-environment. We identified 23 simple sequence repeat (SSR) markers that were significantly associated with the seed oil content. Comparative analysis revealed that five QTL identified in the DH population, located on chromosomes A03, A09, A10 and C09, were co-localized with 11 significantly associated SSR markers that were identified from the association mapping population. Of which, the QTL on chromosome A10 was found to be homeologous with the QTL on chromosome C09 by aligning QTL confidence intervals with the reference genomes B. napus. Those QTL associated with specific macro-environments provides valuable insight into the genetic regulation of seed oil content and will facilitate marker-assisted breeding of B. napus.     

Identification and confirmation of quantitative trait loci for stachyose content in soybean seed

Stachyose is an unfavorable sugar in soybean meal that causes flatulence for non‐ruminant animals. Understanding the genetic control of stachyose in soybean will facilitate the modification of stachyose content at the molecular level. The objective of this study was to identify quantitative trait loci (QTL) associated with seed stachyose content using simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers. A normal stachyose cultivar, ‘Osage’, was crossed with a low stachyose line, V99‐5089, to develop a QTL mapping population. Two parents were screened with 33 SSR and 37 SNP markers randomly distributed on chromosome 10, and 20 SSR and 19 SNP markers surrounding a previously reported stachyose QTL region on chromosome 11. Of these, 5 SSR and 16 SNP markers were used to screen the F_3:4 lines derived from ‘Osage’ x V99‐5089. Seed samples from F_3:5 and F_3:6 lines were analyzed for stachyose content using high‐performance liquid chromatography (HPLC). Composite interval mapping analysis indicated that two stachyose QTL were mapped to chromosome 10 and 11, explaining 11% and 79% of phenotypic variation for stachyose content, respectively. The SSR/SNP markers linked to stachyose QTL could be used in breeding soybean lines with desired stachyose contents. Chi‐square tests further indicated that these two QTL probably represent two independent genes for stachyose content. Therefore, a major QTL was confirmed on chromosome 11 and a novel QTL was found on chromosome 10 for stachyose content.     

基于两个陆地棉低世代群体定位纤维品质相关QTL

【目的】定位棉花纤维品质性状相关的数量性状位点(Quantitative trait locus,QTL)。【方法】以陆地棉高强纤维品系中棉所679和纤维品质一般的农垦5号为亲本构建包含200个单株的F2群体及对应的F2:3家系群体,对2个群体的纤维长度、断裂比强度等5个纤维品质性状进行检测。用6 688对简单重复序列(Simple sequence repeat, SSR)引物在双亲间筛选,得到149对多态性引物,以F2为作图群体,使用QTL IciMapping软件进行连锁图谱构建,并对F2及F2:3群体进行QTL定位。【结果】根据F2群体基因型信息构建了1张包含119个标记、28个连锁群、总长为1 173.5 cM(centiMorgan)的遗传连锁图谱。分别在F_2、F2:3群体中检测到9个和11个与纤维品质性状相关的QTLs,这些QTLs分布在11个连锁群上。其中F2群体的qFL-D11-1、q BT-D11-1与F2:3群体的qFL-D11-1、q MIC-D11-1均定位在标记DPL0062与HAU0423之间,推测这些位点可能是控制纤维品质性状的重要QTL。【结论】利用多个群体进行QTL定位有益于发现稳定的QTL位点,控制纤维品质性状的基因可能成簇存在,为挖掘纤维品质性状相关基因及分子标记辅助育种奠定基础。