Development and mapping of gene-tagged SNP markers in laccases of maize (Zea mays L.)

Laccases, EC 1.10.3.2 or p -diphenol : dioxygen oxidoreductases, have been proposed to be involved in the oxidative polymerization of monolignols into lignins in plants. While 17 laccases have been identified in Arabidopsis , only five ( ZmLac1–5 ) have so far been identified in maize. By a bioinformatic approach, 14 putative laccases were identified in maize. One putative laccase was identical to ZmLac1 , while five were highly homologous to either ZmLac4 or ZmLac5 . Sequence alignment of allelic sequences enabled the development of TaqMan single nucleotide polymorphism (SNP) markers for nine putative laccases. Four of these gene-tagged SNP markers were validated in a doubled-haploid mapping population of 140 individuals, mapping these loci to chromosomes 1, 2, 3 and 7, respectively.     

基于高通量测序开发玉米高效KASP分子标记

SNP (Single Nucleotide Polymorphism)在基因组中数量多、分布广,适用于大规模、自动化基因型检测。本研究利用205份不同来源的玉米自交系全基因组重测序数据鉴定出一系列多态性高的二态性SNP位点并开发出700个KASP分子标记。其中, 202个在46个玉米代表系中得到验证的KASP标记进一步用于系统进化树构建及群体结构分析。结果显示,开发成功的KASP标记在染色体上分布均匀,平均PIC为0.463,平均MAF为0.451。基于KASP标记位点和总SNP位点的聚类分析结果高度吻合。KASP标记位点与总SNP位点的遗传距离相似性系数高达89.5%,能成功区分玉米的杂种优势群。该KASP标记可在玉米种质资源分析、连锁群构建以及杂种优势群划分等方面发挥重要作用。     

Optimum Stand Height for Forage Brown Midrib Sorghum × Sudangrass in North-eastern USA

Brown midrib sorghum × sudangrass is attracting attention in the north‐eastern USA because of its ability to produce acceptable forage yields on marginal corn ground. Other potential advantages include conservation of soil on highly erosive ground, the possibility to spread manure mid‐summer (after first cutting) when runoff risk is minimal, greatly reduced needs for herbicides and the potential to reduce the importation of phosphorus onto the farm. Best management practices need to be developed to take full advantage of the crop in the north‐east. Seven field studies were conducted from 2000 to 2002 in four different agricultural regions in New York to determine the time of harvest optimum for yield and quality in one‐ to three‐cut management systems. Yield, rather than a change in forage quality indicators, was the key determining factor for estimated milk production. However, to prevent the shift from vegetative to reproductive growth (and associated decline in quality) and to better manage the amount of water at harvest, we recommend that in the north‐eastern USA brown midrib sorghum sudangrass be managed using a two‐cut system with harvest taking place when stand heights are about 125 cm or less.     

玉米穗轴粗全基因组关联分析

玉米穗轴粗是一个影响玉米产量和穗轴产量的重要性状,其遗传机制的解析可以对高产育种提供指导。本研究以309份玉米自交系为材料,利用测序基因型分型技术对其进行基因型鉴定。采用FarmCPU(fixed and random model circulating probability unification)、MLMM (multiple loci mixed linear model)和CMLM (compressed mixed linear model)方法,对2017年和2019年河南原阳、河南郸城、河南虞城、海南三亚以及最佳线性无偏估计值环境的穗轴粗,进行全基因组关联分析,共鉴定12个与穗轴粗显著关联的SNP (single nucleotide polymorphisms)(P<8.60E-07)。其中,S4₂9277313利用FarmCPU和MLMM方法在2017年原阳均检测到。S1₂9006330、S2₁70889116、S2₂046026464和S4₈3821...     

几种分子标记技术在玉米自交系类群划分中的应用

分子标记技术的开发利用推动了玉米育种的发展,概述了5种常用分子标记SSR、RFLP、RAPD、AFLP、SNP的原理及特点。综述了分子标记技术在玉米自交系类群划分中的应用。     

基于全基因组关联分析解析玉米籽粒大小的遗传结构

玉米籽粒大小是产量重要构成因子之一,也是受多基因调控的复杂数量性状,挖掘玉米籽粒大小相关性状的关键调控基因,将有助于提高玉米的产量。本研究以212份优良玉米自交系为材料,于2018年和2019年分别对粒长、粒宽和粒厚进行测定,并结合均匀分布于玉米基因组的73,006个单核苷酸多态性(single nucleotide polymorphism,SNP)标记进行全基因组关联分析。基于FarmCPU算法,在玉米的10条染色体上检测到47个与籽粒大小相关性状关联的SNP。结合B73玉米自交系籽粒发育的动态时空转录数据,在显著SNP标记的连锁不平衡区域内,共检测到58个与籽粒大小相关的候选基因,其编码的蛋白与多种蛋白存在互作关系,参与并调控多个与籽粒发育密切相关的生物学过程。本研究为解析玉米籽粒发育的分子调控机制,改良籽粒大小和提高作物产量提供了新的参考。     

QTL mapping of improving forage maize starch degradability in European elite maize germplasm

Improving maize starch content is of great importance for both forage and grain yield. In this study, 13 starch degradability traits were analysed including percentage of the seedling area, floury endosperm, hard endosperm of total grain area, percentage of the floury endosperm surface and vitreousness ratio surface hard: floury endosperm surface, etc. We mapped quantitative trait loci (QTL) in a biparental population of 309 doubled haploid lines based on field phenotyping at two locations. A genetic linkage map was constructed using 168 SSR (simple sequence repeat) markers, which covered 1508 cM of the maize genome, with an average distance of 9.0 cM. Close phenotypic and genotypic correlations were found for all traits, and were all statistically significant (p = 0.01) at two locations. Major QTL for more than two traits were detected, especially in two regions in bins 4.05–4.06 and 7.04–7.05, associated with 13 and 9 traits, respectively. This study contributes to marker‐assisted breeding and also to fine mapping candidate genes associated with maize starch degradability.     

Genome‐wide identification of candidate phosphate starvation responsive genes and the development of intron length polymorphism markers in maize

To develop genetic markers associated with tolerance to low phosphorus, we identified candidate phosphate starvation responsive (PSR) genes and developed their intron length polymorphism (ILP) markers in maize on a genome‐wide scale. Based on the known plant PSR genes, 161 candidate PSR genes were identified. Of these genes, 138 genes contained at least one intron and were then used to develop 606 PSR‐ILP markers by designing PCR primers to the intron‐flanking exonic regions. PCR evaluation was performed on 43 randomly selected PSR‐ILP markers in 30 maize inbred lines. Of the primers, 88.4% amplified stable and pure products in all maize inbred lines, and 53.5% of the markers showed ILP, with PIC values ranging from 0.06 to 0.80. The result of clustering analysis of the 30 maize inbred lines, based on the polymorphism of 23 ILP markers, suggests that the ILP markers developed in this study are not only efficient for genetic diversity analysis but also potentially useful for marker assisted selection for tolerance to low phosphorus.     

玉米Suwan种质改良过程中的关键基因组区段发掘

玉米Suwan种质抗性好、适应性强、籽粒品质优,在现代育种尤其是南方玉米育种中具有不可替代的作用。明确Suwan种质优良特性在改良过程中的遗传机制对我国南方玉米生态区的玉米生产具有重要意义。本研究以Suwan1(Suwan1C10)及其衍生群体(苏兰1号C0)不同改良世代为材料,利用包含5.6万个SNP标记的MaizeSNP50芯片对供试群体进行基因型鉴定。遗传分析发现:Suwan 1群体不同改良世代间的基因组差异片段较少,仅出现5个,其中4个出现在第11轮改良世代(Suwan 1 C11), 1个出现在第15轮改良世代(Suwan 1 C15);苏兰1号不同改良世代间的基因组差异片段相对较多,共有18个,其中8个在不同改良世代间稳定遗传; Suwan种质改良形成苏兰1号群体的过程中,共获得43个Lancaster特异性遗传片段,其中35个在苏兰1号不同改良世代间稳定遗传。全基因组关联分析共鉴定出16个与穗行数显著关联的QTN,分别位于第2、第3、第5、第6、第7、第8、第9染色体上,其中SYN25713和SYN36577位于苏兰1号群体的Lancaster特异性遗传片段内;共检测到13个控制穗长相关的QTN,分别位于第1、第2、第5、第7、第8、第9染色体上,其中PZE-105143697位于苏兰1号群体的Lancaster特异性遗传片段内。该结果为后续全基因组关联研究和分子标记辅助选择等提供了重要的理论依据。     

Genetic characterization of 218 elite CIMMYT maize inbred lines using RFLP markers

Characterization of genetic diversity among maize inbred lines can facilitate organization of germplasm and improve efficiency of breeding programs. A set of 218 phenotypically diverse inbred maize lines developed at CIMMYT for hybrid production was characterized using 32 RFLP markers to: (1) analyze the genetic diversity present; (2) define potential heterotic groups based on clusters formed with marker data; and (3) identify the most representative testers for each potential heterotic group. Lines were clustered using five different genetic distance measurements to find consensus non-hierarchical clusters. Dendrograms were produced to study hierarchical classification within smaller groups of lines. A very high average allelic diversity was seen in this germplasm. Lines did not cluster based on phenotype, environmental adaptation, grain color or type, maturity, or heterotic response (as determined based on hybrid performance with testers), but lines related by pedigree usually did cluster together. Previously defined testers from opposite heterotic groups were not genetically differentiated, and did not represent well their heterotic group. Discrete clusters were difficult to find; thus, potential heterotic groups will be difficult to suggest using RFLP markers alone. However, suggestions on how to use molecular markers and cross performance information to refine heterotic groups and select representative testers are presented.     

Relationship between genetic markers and morphological traits in a maize inbred lines collection

Summary Genetic markers (isozymes), in addition to the traits recommended by the UPOV convention, have been used in France since 1989 to characterize the maize inbred lines submitted to registration. In the years 1989. 1990 and 1991, a total of 974 inbreds has been described according to this procedure. Relationships between genetic markers were investigated and underlined the occurrence of linkage disequilibria within the tested germplasm. These disequilibria appeared to depend strongly on the breeding history of the germplasm. In some cases, these disequilibria could be related to a major progenitor (foundation effect). Relationships between genetic markers and quantitative traits were also pointed out. High coefficients of determination (up to 60%) were in some cases observed at the within group level, and also appeared to be very dependent on the major progenitors of the group of interest. The consequences of these results for breeding and distinctiveness studies are discussed.     

Reciprocal differences for forage traits in single and three-way crosses of maize

Eleven flint inbreds of maize were crossed with 11 dent inbreds according to a balanced incomplete factorial mating design to produce 66 single crosses (2W) in both reciprocal forms. Additionally, six three-way crosses (3W) were also produced in both reciprocal forms. These hybrids were evaluated for 11 forage yield and quality traits in 1983 and 1984 at three sites in Germany. Highly significant reciprocal differences were observed for nine traits in the 2W. The respective variance component estimates amounted to 2–33% of the variance component due to crosses. On average, flint × dent 2W had lower dry-matter content (DMC) than dent × flint 2W and a 1 % greater dry-matter yield (DMY) of forage. Three-way crosses produced on singles exhibited greater DMC (3%) and DMY (5%) of the ear, metabolizable energy content (MEC) of stover (1%), and DMC and DMY of forage (2%) than their reciprocal forms produced on lines. Based on this study, because of their minor importance, routine testing for reciprocal differences among 2W hybrids is not recommended.     

基于高通量测序的大麦In Del标记开发及应用

分子标记是遗传研究的基础工具,广泛应用于遗传多样性研究、种质鉴定、遗传图谱构建和基因定位等领域。本研究基于大麦的全基因组重测序数据鉴定出的二态性SNP位点,开发出遍布全基因组的118对InDel引物。以49份不同地理来源的大麦种质检测其有效性,筛选出2个等位基因的共显性InDel标记72对,进一步对288份大麦种质进行遗传距离分析及构建系统发育树。结果显示,筛选到32个有效性的核心InDel标记,覆盖大麦7条染色体上,平均PIC为0.44,平均MAF为0.34;基于InDel标记位点的供试大麦的系统发育树结果揭示供试大麦具有丰富的遗传多样性,且能够将具有相同地理来源的多数品种聚为一类。表明开发的32个二态性InDel标记可有效地用于鉴定品种间的亲缘关系,且丰富了大麦品种鉴定的分子标记。上述核心InDel标记在大麦品种鉴定、大麦资源亲缘关系分析以及群体划分方面具有一定的理论意义和应用价值。     

Heterotic groups based on yield-specific combining ability data and phylogenetic relationship determined by RAPD markers for 28 tropical maize open pollinated varieties

Twenty eight maize open pollinated varieties (OPVs) were crossed in a diallel scheme and the 378 F₁'s were evaluated in 10 environments in Brazil. Based on yield-specific combining ability data (SCA), these varieties were classified in four heterotic groups. The consistence of the proposed heterotic groups was confirmed comparing intra- and inter-group F₁ values and midparent heterosis. Superior OPVs combinations for use as a source of inbreds in hybrid breeding programs were determined. RAPD markers were used to genotype these varieties. A UPGMA dendogram, based on marker data from 50 primers and 178 polymorphic bands, was obtained. Phylogeny obtained with RAPD markers agreed with known pedigree data. Dent germplasm tended to group separately from flint germplasm. Multidimensional Scaling Analysis on marker data and morphological data showed a higher degree of genetic divergence among the dent germplasm than among the flint germplasm used in this study. Correlation between RAPD marker estimated genetic distance and SCA for yield was low and positive (r = 0.16^**). This revised version was published online in August 2006 with corrections to the Cover Date.     

Isolation and functional analysis of the zmARM4 locus in a novel maize (Zea mays) grain-filling mutant

Maize grain filling (GF) and grain weight (GW) are closely associated with grain yield. In our study, the ZmARM4 locus affecting GF and GW was identified using a reverse genetics screen of UniformMutator (Mu). First, MutatorDR-flanking sequence (MuDRFs) was isolated from the mutant by the MuDR-amplified fragment length polymorphism method, and it was verified by cosegregation (between MuDRFs and phenotype) and linkage analyses. Second, the target gene of the MuDRFs was found to encode the Armadillo domain of the repeat protein gene families and has high expression in the GF stage. Third, association mapping between SNPs in the ZmARM4 region and the GW of 489 maize lines indicated that ZmARM4 is a functional gene that is closely related to GW. Moreover, bioinformatics analysis suggested that ZmARM4 may be relevant for the activity of sucrose transport or the number of cell divisions in endosperm cells. Therefore, ZmARM4 may be used as an important locus for maize GW in molecular breeding.     

近红外分析技术在玉米种质资源品质分析中应用

近红外分析技术具有快速、简便、准确、非破坏性的优点,为玉米品质育种提供了一个新的技术手段。利用FOSS公司1255型近红外透射光谱分析仪,对2000 年～2001年的686份玉米自交系和2001年～2002年的1438份杂交组合的籽粒品质成分进行测定。结果表明,玉米籽粒3种品质成分在不同材料间存在显著差异,粗脂肪和蛋白质变异幅度和变异系数都较大,粗淀粉含量变异幅度和变异系数都较小,年份之间有差异。自交系中粗脂肪平均含量分别为4.24%和4.27%,蛋白质平均含量分别为11.75%和12.17%,粗淀粉平均含量分别为71.04%和71.10%,玉米杂交组合粗脂肪平均含量分别为4.53%和4.86%,蛋白质平均含量分别为9.93%和10.05%,粗淀粉平均含量分别为72.56%和72.74%。     

玉米叶片紫色中脉性状的遗传

利用引进的紫黑色种皮玉米随机交配群体,通过杂交和多代单株自交选择,培育出紫色中脉稳定纯系06Z-2。以06Z-2(P₂)和白色中脉自交系L-24(P₁)为材料,创制出F₁、F₂、BC₁和BC₂群体,对玉米叶片紫色中脉性状的遗传特征进行了研究。结果表明,紫色中脉性状属于显性单基因遗传,分离世代中紫色中脉株数与白色中脉株数的分离比符合孟德尔经典遗传学第一定律,且遗传稳定,易于转育。深入开展玉米叶片紫色中脉性状的遗传与应用研究,对于玉米自交系或杂交种防杂保纯、进行种子纯度鉴定、保护知识产权等具有重要的理论与适用价值。     

糯玉米自交系SSR标记遗传多样性及群体遗传结构分析

为了解糯玉米种质的遗传基础,利用29对SSR标记对87份糯玉米自交系进行遗传多样性分析,共检测出180个等位变异,平均每个位点6个等位变异,多态性信息含量变幅为0.308～0.915,平均为0.572。材料间遗传相似系数为0.49～0.93,平均为0.66。通过聚类分析UPGMA (unweighted pair-group method with arithmetic means)方法在遗传相似系数0.64处将87份糯玉米自交系划分为4个类群,分别包含9、66、10和2份材料。此外,利用Structure群体遗传结构分析也将87份糯玉米自交系分为4个类群,分别包含24、25、19和19份材料;进一步分析表明,供试群体中大部分糯玉米自交系的遗传变异较单一。本研究为糯玉米新品种选育和遗传进化分析提供了种质基础和理论依据。     

玉米抗倒伏相关性状QTL的关联和连锁分析

倒伏是影响玉米高产和机械化收获的重要因素,明确玉米抗倒伏相关性状的遗传基础,增强玉米品种抗倒伏能力,可为玉米高产宜机收育种提供理论依据。本研究以国内外收集的153份自交系为材料,利用6H90K SNP芯片检测的70,438个高质量SNP标记对地上第3节茎秆强度、株高、穗位高和穗位系数进行全基因组关联分析,分别检测到与茎秆强度、株高、穗位高和穗位系数相关位点5个、14个、16个和21个,单个关联位点最大效应值为13.24。同时以KA105/KB020的F₅群体为试验材料,采用QTL IciMapping V4.2软件的完备区间作图法进行QTL定位,检测到21个与抗倒伏相关的QTL,可解释表型变异3.86%～16.58%。结合关联分析和连锁分析结果发现, 2个QTL区间与关联分析的候选区间重合。经过候选区段的基因功能注释和文献查阅,挖掘到GRMZM2G105391、GRMZM2G014119和GRMZM2G341410等与细胞壁生物合成、细胞分裂和细胞伸长的相关基因可供进一步分析。本研究结果为进一步解析玉米抗倒伏性状的遗传基础提供参考。