Quantitative trait loci analysis in Theobroma cacao using molecular markers. Inheritance of polygenic resistance to Phytophthora palmivora in two related cacao populations

Two related segregating populations of Theobroma cacao L. were analysed for their resistance to Phytophthora palmivora. The first F₁ population was obtained by crossing two susceptible cacao clones of Catongo (a highly homozygous genotype) and Pound 12(a highly heterozygous genotype) and the second population was obtained by backcrossing a single F₁ tree with Catongo. The genetic maps obtained for each population were compared. The F₁ map includes 162 loci and the backcross has 140 loci. The two maps, F₁ and BC₁, exhibit high co-linear loci organization covering respectively, 772 and 944 cM.Phytophthora resistance was assessed by measuring the size increase of a lesion at five (DL5)and ten days (DL10) after pod inoculation. Six different QTL were detected in the F₁ and BC₁ populations. One QTL was found in both populations, and appeared to be a major component of disease resistance, and explaining nearly 48% of the phenotypic variance in the F₁ population. The absence of some yield QTL detection in the BC₁ in comparison with the F₁ population is due to the lack of transmission of the favouring alleles for these QTL from the single F₁ tree used for the backcross. The phenotypic variance explained by the action of the quantitative trait alleles indicated that genetic factors of both major and minor effects were involved in the control of the character studied. QTL conferring increased resistance to Phytophthorawere identified in both susceptible parents, suggesting the presence of transgressive traits and the possibility of selection in cacao. Pleiotropic and epistatic effects for the QTL were also detected. Finally, the use of marker assisted selection (MAS) in cacao breeding programs is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

QTL mapping of maize (Zea mays) stay-green traits and their relationship to yield

A maize genetic linkage map derived from 115 simple sequence repeat (SSR) markers was constructed from an F₂ population. The F₂ was generated from a cross between a stay-green inbred line (Q319) and a normal inbred line (Mo17). The map resolved 10 linkage groups and spanned 1431.0 cM in length with an average genetic distance of 12.44 cM between two neighbouring loci. A total of 14 quantitative trait loci (QTL) were detected for stay-green traits at different postflowering time intervals and identified by composite interval mapping. The respective QTL contribution to phenotypic variance ranged from 5.40% to 11.49%, with trait synergistic action from Q319. Moreover, maize stay-green traits were closely correlated to grain yield. Additional QTL analyses indicated that multiple intervals of stay-green QTL overlapped with yield QTL.     

陆地棉衣分差异群体产量及产量构成因素

 以衣分差异较大的陆地棉品种为材料,构建了包含188个F₂单株的作图群体,应用6111对SSR引物对亲本进行了分子标记筛选,结果仅获得了123个多态性位点,其中88个位点构建了总长为666.7 cM、平均距离为7.57 cM的遗传图谱,覆盖棉花基因组的14.9%。通过复合区间作图法对F₂单株和F_2∶3家系进行QTL检测,共鉴定出了18个控制产量及产量构成因素变异的QTLs,包括2个衣分QTLs、4个子棉产量QTLs、4个皮棉产量QTLs、2个衣指QTLs、3个单株铃数QTLs、2个铃重QTLs和1个子指QTL。 解释的表型变异分别为\{6.9%\}～16.9%、5.6%～16.2%、4.8%～15.6%、7.7%～13.3%、8.2%～11.6%、6.1%～7%和6.6%。不同QTLs在相同染色体区段上的成簇分布表明与产量性状相关的基因可能紧密连锁或一因多效。产量及产量构成因素QTLs的遗传方式主要以显性和超显性效应为主。检测到的主效QTLs可以用于棉花产量及产量构成因素的分子标记辅助选择。     

Quantitative trait loci mapping and epistatic analysis for grain yield and yield components using molecular markers with an elite maize hybrid

Summary The aim of this investigation was to map quantitative trait loci (QTL) associated with grain yield and yield components in maize and to analyze the role of epistasis in controlling these traits. An F_2:3 population from an elite hybrid (Zong3 × 87-1) was used to evaluate grain yield and yield components in two locations (Wuhan and Xiangfan, China) using a randomized complete-block design. The mapping population included 266 F_2:3 family lines. A genetic linkage map containing 150 simple sequence repeats and 24 restriction fragment length polymorphism markers was constructed, spanning a total of 2531.6 cM with an average interval of 14.5 cM. A logarithm-of-odds threshold of 2.8 was used as the criterion to confirm the presence of one QTL after 1000 permutations. Twenty-nine QTL were detected for four yield traits, with 11 of them detected simultaneously in both locations. Single QTL contribution to phenotypic variations ranged from 3.7% to 16.8%. Additive, partial dominance, dominance, and overdominance effects were all identified for investigated traits. A greater proportion of overdominance effects was always observed for traits that exhibited higher levels of heterosis. At the P ≤ 0.005 level with 1000 random permutations, 175 and 315 significant digenic interactions were detected in two locations for four yield traits using all possible locus pairs of molecular markers. Twenty-four significant digenic interactions were simultaneously detected for four yield traits at both locations. All three possible digenic interaction types were observed for investigated traits. Each of the interactions accounted for only a small proportion of the phenotypic variation, with an average of 4.0% for single interaction. Most interactions (74.9%) occurred among marker loci, in which significant effects were not detected by single-locus analysis. Some QTL (52.2%) detected by single-locus analysis were involved in epistatic interactions. These results demonstrate that digenic interactions at the two-locus level might play an important role in the genetic basis of maize heterosis.     

Identification of QTL for oil content,seed yield,and flowering time in oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>)

A genetic map was constructed with 353 sequence-related amplified polymorphism and 34 simple sequence repeat markers in oilseed rape (Brassica napus L.). The map consists of 19 linkage groups and covers 1,868 cM of the rapeseed genome. A recombinant doubled haploid (DH) population consisting of 150 lines segregating for oil content and other agronomic traits was produced using standard microspore culture techniques. The DH lines were phenotyped for days to flowering, oil content in the seed, and seed yield at three locations for 3 years, generating nine environments. Data from each of the environments were analyzed separately to detect quantitative trait loci (QTL) for these three phenotypic traits. For oil content, 27 QTL were identified on 14 linkage groups; individual QTL for oil content explained 4.20–30.20% of the total phenotypic variance. For seed yield, 18 QTL on 11 linkage groups were identified, and the phenotypic variance for seed yield, as explained by a single locus, ranged from 4.61 to 24.44%. Twenty-two QTL were also detected for days to flowering, and individual loci explained 4.41–48.28% of the total phenotypic variance.     

甘蓝型油菜产量及其构成因素的QTL定位与分析

产量性状是复杂的数量性状, 对种子的单株产量及其构成因素(全株总有效角果数、每角粒数、千粒重)进行QTL定位和上位性分析,确定其在染色体上的位置及其遗传效应,可以探讨油菜杂种优势产生原因,提高育种中对产量性状优良基因型选择的效率,达到提高油菜产量的目的。在双低油菜细胞质雄性不育保持系1141B和双高恢复系垦C₁构建的F₂作图群体中,运用SRAP、AFLP和SSR三种标记技术构建了一个甘蓝型油菜(Brassica napus L.)的分子标记遗传连锁图谱。共包含244个标记,分布于20个主要连锁群、1个三联体上,图谱总长度为2 769.5 cM。采用Windows QTL Cartographer Version 2.0统计软件及复合区间作图法,对油菜单株产量及其3大构成因素进行QTL定位,共检测到QTLs 16个分布在9个连锁群上,其中第6和13连锁群最多,均有3个。单个QTL解释性状表型变异的0.38%～73.34%。对于同一性状,等位基因的增效作用既来自母本,亦源自父本;采用双向方差分析法对位点间互作及其上位性进行分析,检测到26对影响产量构成性状的上位性互作效应QTL,说明油菜基因组中存在大量控制产量的互作位点,油菜产量性状的上位性存在着多效性,上位性互作包括QTL与非QTL位点,其中以非QTL位点较多。一般互作位点的独立效应值较小,而互作的效应值显著增大,且一般超过两位点独立效应值之和。反映了控制产量性状基因的复杂性。上位性是甘蓝型油菜产量性状杂种优势的重要遗传基础。     

干旱和灌溉两种处理条件下玉米株高、产量的QTL分析

为了对玉米的抗旱遗传学研究以及开展抗旱分子标记辅助育种提供有力支撑,利用SSR分子标记技术,构建了玉米基于RIL6群体的遗传连锁图谱,包含101个位点,覆盖玉米基因组1 395.2 cM,标记间平均距离为13.81 cM。在灌溉与干旱两种环境下,对该RIL家系的株高、产量进行QTL初级定位,在灌溉条件下,检测到3个控制株高和3个控制产量的QTL,在干旱条件下,检测到5个控制株高和3个控制产量的QTL,两种水分条件下检测出的QTL不同,并找到1个相对稳定的QTL。     

甘蓝型油菜产量及相关性状的QTL分析

高产是甘蓝型油菜育种的重要目标之一，产量是多基因控制的数量性状。本文通过QTL作图分析了产量及其相关性状的数量性状位点，以甘蓝型油菜中油821和保604 F₁代小孢子培养获得的DH系为作图群体，构建了由20个连锁群组成的，包括251个分子标记( 2个RFLP标记，72个RAPD标记，91个SSR标记，86个SRAP标记)的遗传连锁图(10个标记没有分配到连锁群中)。图谱的平均图距为6.96 cM，共覆盖油菜基因组1 746.5 cM。在此图谱基础上采取复合区间作图法，检测到与油菜产量及其相关性状有关的QTL共17个。其中控制株高的3个分别位于第4、第9和第10连锁群上，对性状的解释率为9.42%～17.58%；与分枝部位有关的4个分别位于第4、第6和第7连锁群上，其中Bp1 和Bp2 均位于第4连锁群，对性状的解释率为8.13%～15.20%；与主花序有效长有关的3个分别位于第4、第10和第16连锁群上，对性状的解释率为7.49%～23.36%；与一次有效分枝有关的2个分别位于第1、第4连锁群上，对性状的解释率为15.29%～19.58%；与角果总数和千粒重有关的分别位于第4连锁群和第9连锁群上，贡献率分别为17.42%和7.64%；与单株产量有关的3个分别位于第3、第4和第15连锁群，共解释26.60%的表型变异。部分性状的QTL在连锁群上成簇分布,对性状贡献率很大，表现主效QTLs的特点，相应的性状之间也呈显著相关，这表明一因多效或者相关的QTLs之间紧密连锁是性状相关的遗传基础。本研究中与主效QTLs连锁的标记可用于油菜产量性状的分子标记辅助选择。     

Genome mapping and molecular markers identification for yield,yield component and fibre quality traits in tetraploid cotton

The identification of quantitative trait loci (QTL) across different environments is a prerequisite for marker‐assisted selection (MAS) in crop improvement programmes. CottonSNP63k Illumina infinium array was used for genotyping 178 inter‐specific recombinant inbred lines and the parents, and identified 1,667 homozygous polymorphic markers between the parents. Of these, 1,430 markers were used for the construction of linkage map after removing 237 redundant markers. The genetic map spans a total genetic length of 3,149.8 cM with an average marker interval size of 2.2 cM. The phenotypic data from five environments were analysed separately using inclusive composite interval mapping which identified a total of 56 QTL explaining phenotypic variances (PVE) in the range of 8.18%–28.91%. There were 11 and 24 major QTL found for fibre quality and yield components, respectively. A total of 64 QTL were identified through Multi‐Environment Trials analysis, of which 34 recorded QTL × Environment interactions.     

苦荞SSR分子遗传图谱的构建及分析

构建苦荞遗传连锁图谱,为今后有关苦荞基因组结构、重要农艺性状QTL定位、分子标记辅助育种和基因克隆等研究工作奠定基础。以栽培苦荞‘滇宁一号’和苦荞野生近缘种杂交产生的119份F4代分离材料为作图群体,利用SSR分子标记来构建苦荞的分子遗传连锁图谱。本研究构建的连锁图谱包含15个连锁群,由89个标记组成,其中偏分离的标记有22个,占24.7%,每条连锁群上的标记在2~16之间。连锁群长度在6.9~165.8 cM的范围,覆盖基因组860.2 cM,总平均长度9.7 cM。本研究构建了首张苦荞SSR遗传连锁图谱,为苦荞QTL定位、基因克隆、遗传选育等研究奠定了基础。     

栽培种花生荚果大小相关性状QTL定位

以远杂9102为母本,徐州68-4为父本杂交衍生的F5和F6共188个家系,构建了一张包含365个标记,总长度713.07 c M,标记间平均距离1.96 c M的栽培种花生遗传图谱。图谱包含22个连锁群,各连锁群平均长度12.37~81.39 c M,连锁群上标记数量3~46个。结合2013和2014年采集的荚果表型数据,采用Win QTLcart 2.5软件的复合区间作图法(composite interval mapping,CIM)进行QTL定位和效应估计。2个环境下共检测到41个QTL,其中与荚果长、宽、厚和百果重相关的QTL分别为13、7、13和8个,表型变异解释率为3.14%~18.27%。有6个QTL在2种环境下被重复检测到,其中百果重相关的2个(q HPWLG13.1、q HPWLG14.1),分布在LG13和LG14连锁群,遗传贡献率为6.95%~14.60%;与荚果长相关的3个(q LPLG2.2、q LPLG13.1、q LPLG14.1),分布在LG2、LG13和LG14连锁群,遗传贡献率为3.14%~18.27%;与荚果厚相关的1个(q TPLG3.4),分布在LG3连锁群,遗传贡献率为8.24%~9.24%。本研究涉及性状存在9个QTL热点区,每个热点区涉及2~3个性状,表型贡献率为3.57%~18.27%。     

普通小麦多酚氧化酶活性的QTL分析

多酚氧化酶（polyphenol oxidase, PPO）是引起面团（片）颜色褐变的主要原因。利用122对SSR引物、4对贮藏蛋白的STS引物和10对AFLP引物组合，分析了中优9507´CA9632的71个DH系的基因型，构建了由173个位点组成的遗传连锁图，在小麦21个连锁群上覆盖2 881 cM。将该群体种植于3种环境，采用复合区间作图法（CIM）进行了P     

Genetic mapping of quantitative trait loci for fiber quality and yield trait by RIL approach in Upland cotton

The improvement of cotton fiber quality has become more important because of changes in spinning technology. Stable quantitative trait loci (QTLs) for fiber quality will enable molecular marker-assisted selection to improve fiber quality of future cotton cultivars. A simple sequence repeat (SSR) genetic linkage map consisting of 156 loci covering 1,024.4 cM was constructed using a series of recombinant inbred lines (RIL) developed from an F₂ population of an Upland cotton (Gossypium hirsutum L.) cross 7235 × TM-1. Phenotypic data were collected at Nanjing and Guanyun County in 2002 and 2003 for 5 fiber quality and 6 yield traits. We found 25 major QTLs (LOD ≥ 3.0) and 28 putative QTLs (2.0 < LOD < 3.0) for fiber quality and yield components in two or four environments independently. Among the 25 QTLs with LOD ≥ 3, we found 4 QTLs with large effects on fiber quality and 7 QTLs with large effects on yield components. The most important chromosome D8 in the present study was densely populated with markers and QTLs, in which 36 SSR loci within a chromosomal region of 72.7 cM and 9 QTLs for 8 traits were detected.     

Molecular markers for yield components in Brassica juncea – do these assist in breeding for high seed yield?

A population of 112 F₁-derived doubled haploid lines was produced from a reciprocal cross of Brassica juncea. The parents differed for seed quality, seed color and many agronomic traits. A detailed RFLP linkage map of this population, comprising 316 loci, had been constructed, and was used to map quantitative trait loci (QTL) for seed yield and yield components, viz. siliqua length, number of seeds per siliqua, number of siliques per main raceme and 1000-seed weight. Stable and significant QTLs were identified for all these yield components except seed yield. For yield components, a selection index based on combined phenotypic and molecular data (QTL effects) could double up the efficiency of selection compared to the expected genetic advance by phenotypic selection. Selection indices for high seed yield, based on the phenotypic data of yield and yield components, could only improve the efficiency of selection by 4% of the genetic advance that can be expected from direct phenotypic selection for yield alone. Inclusion of molecular data together with the phenotypic data of yield components in the selection indices did not improve the efficiency of selection for higher seed yield. This is probably due to often negative relationships among the yield components. Most of the QTLs for yield components were compensating each other, probably due to linkage, pleiotropy or developmentally induced relationships among them. The breeding strategy for B. juncea and challenges to marker assisted selection are discussed.     

Construction of a linkage map for quantitative trait loci associated with economically important traits in creeping bentgrass (Agrostis stolonifera L.)

Creeping bentgrass (Agrostis stolonifera L.) is the most widely cultivated and high-value turfgrass species. Genetic linkage maps of creeping bentgrass were constructed for quantitative trait loci (QTL) analysis of gray snow mold (Typhula incarnata) resistance, recovery and leaf width. A segregating population of 188 pseudo-F₂ progeny was developed by two-way pseudo-testcross mapping strategy. Amplified fragment length polymorphism, new developed Agrostis specific expressed sequence tag-single sequence repeat (SSR), random amplified polymorphic DNA and genomic SSR markers corresponding to DNA polymorphisms heterozygous in one parent and null in the other, were scored and placed on two separate genetic linkage maps, representing each parent. In the male parent map, 100 markers were mapped to 14 linkage groups covering a total length of 793?cM with an average interval of 8.2?cM. In the female parent map, 146 markers were clustered in another 14 linkage groups spanning 805?cM with an average distance of 5.9?cM between adjacent markers. We identified three putative QTL for leaf width and one QTL for snow mold disease resistance. The construction of a linkage map and QTL analysis are expected to facilitate the development of disease resistant creeping bentgrass cultivars by using molecular marker-assisted selection.     

不同遗传背景下陆地棉衣分和子指性状QTL定位

为陆地棉产量性状有关的分子标记辅助育种奠定理论基础,以高品质中长绒棉品种‘新陆早24号’为父本,转基因抗虫棉常规品种‘鲁棉研28号’和高产、优质棉花新品种‘冀棉516’为母本,构建F2和F2:3分离群体;利用7638对SSR引物对‘鲁棉研28号’和‘新陆早24号’进行多态性进行筛选,共获得225对多态性引物,对238个F2单株DNA扩增获得238个多态性标记位点,其中185个构建了包括44个连锁群,总长为1509.38 cM的遗传连锁图谱,标记间的平均距离为8.16 cM,覆盖棉花总基因组的33.91%。根据已有图谱的定位结果,40个连锁群与染色体建立联系。利用复合区间作图法定位‘鲁棉研28号’与‘新陆早24号’分离群体F2单株和F2:3家系的衣分和子指性状QTL,其中得到3个衣分和5个子指的QTL;根据定位结果,选择了14对SSR引物,分析‘冀棉516’与‘新陆早24号’的多态性,其中6个标记构建了两个连锁群。1个衣分和1个子指的QTL在两个群体中均检测到,这些共同QTLs为分子标记辅助育种奠定了基础。     

Localization of QTL for basal root thickness in <Emphasis Type="Italic">japonica</Emphasis> rice and effect of marker-assisted selection for a major QTL

Root traits are key components of plant adaptation to drought environment. By using a 120 recombined inbred lines (RILs) rice population derived from a cross between IRAT109, a japonica upland rice cultivar and Yuefu, a japonica lowland rice cultivar, a complete genetic linkage map with 201 molecular markers covering 1,833.8 cM was constructed and quantitative trait loci (QTLs) associated with basal root thickness (BRT) were identified. A major QTL, conferring thicker BRT, located on chromosome 4, designated brt4, explained phenotypic variance of 20.6%, was selected as target QTL to study the effects of marker-assisted selection (MAS) using two early segregating populations derived from crosses between IRAT109 and two lowland rice cultivars. The results showed that the flanking markers of brt4 were genetically stable in populations with different genetic backgrounds. In the two populations under upland conditions, the difference between the means of BRT of plants carrying positive and negative favorable alleles at brt4 flanking markers loci was significant. Phenotypic effects of BRT QTL brt4 were 5.05–8.16%. When selected plants for two generations were planted at Beijing and Hainan locations under upland conditions, MAS effects for BRT QTL brt4 were 4.56–18.56% and 15.46–26.52% respectively. The means of BRT for the homozygous plants were greater than that of heterozygous plants. This major QTL might be useful for rice drought tolerance breeding. L. Liu and P. Mu are contributed equally to this work.     

Detection of QTL for forage yield,lodging resistance and spring vigor traits in alfalfa (Medicago sativa L.)

Alfalfa (Medicago sativa L.) is an internationally significant forage crop. Forage yield, lodging resistance and spring vigor are important agronomic traits conditioned by quantitative genetic and environmental effects. The objective of this study was to identify quantitative trait loci (QTL) and molecular markers associated with increased forage yield, resistance to lodging, and spring vigor. A backcross population composed of 128 progeny was developed by crossing the breeding parents DW000577 (lodging susceptible) and NL002724 (lodging-resistant) and back-crossing an individual F₁ plant to the maternal parent (i.e. DW000577). A linkage map of NL002724 was developed based upon the segregation of 236 AFLP, SRAP, and SSR markers among the backcross progeny. The markers were distributed among 14 linkage groups, covering an estimated recombination distance of 1497.6 centiMorgans (cM). Replicated clones of both parents and backcross progeny were evaluated in the field for estimated forage yield, lodging, and spring vigor in Washington and Wisconsin during 2007 and 2008. Significant QTL were found for all three traits. In particular, two QTL for lodging resistance were identified that explained ≥14 % of trait variation, and were significant in all years and locations. Major QTL explaining over 25 % of trait variation for forage yield were detected in multiple environments at two separate locations on chromosome III. Several QTL for spring vigor were located in the same or similar positions as QTL for forage yield, possibly explaining the significant correlation between these traits. Molecular markers associated with the aforementioned QTL were also identified.     

利用重组自交系定位棉花第22染色体短臂的纤维品质性状的QTL

 以TM-1的染色体片段代换系CSB22sh和TM-1杂交,构建了包含104个家系的重组自交系（RILs）群体。利用74对具有多态性位点的引物进行检测,构建了包含61个多态性位点,长度为76.93 cM的遗传图谱,平均标记间距离1.26 cM。利用此遗传图谱结合重组自交系群体4个环境下的5个纤维品质性状进行QTL定位,共定位出12个QTLs,解释性状表型变异的2.45%～21.11%;在1,2,3,4个环境中同时检测到的QTLs分别有9,1,1,1个。而利用4个环境平均值进行联合分析定位出个4个QTLs,纤维长度和纤维整齐度的QTLs均为2个,解释性状表型变异的14.37%～19.97%,并且纤维长度和整齐度的QTL在相同的位置。多环境下检测到的QTL可能对标记辅助选择有实际应用价值。     

Genetic linkage map of Chinese native variety faba bean (Vicia faba L.) based on simple sequence repeat markers

Simple sequence repeat (SSR) marker is a powerful tool for construction of genetic linkage map which can be applied for quantitative trait loci (QTL) and marker‐assisted selection (MAS). In this study, a genetic map of faba bean was constructed with SSR markers using a 129 F₂ individuals population derived from the cross of Chinese native variety 91825 (large seed) and K1563 (small seed). By screening 11 551 SSR primers between two parents, 149 primer pairs were detected polymorphic and used for F₂ population analysis. This SSR‐based genetic linkage map consisted of 15 linkage groups with 128 SSR. The map encompassed 1587 cM with an average genetic distance of 12.4 cM. The genetic map generated in this study will be beneficial for genetic studies of faba bean for identification of marker‐locus‐trait associations as well as comparative mapping among faba bean, pea and grasspea.