Development of molecular linkage maps in sweet potato (Ipomoea batatas L.) using sequence‐related amplified polymorphism markers

Sweet potato is an important food crop with high starch content. It is a hexaploid species, for which the chromosomes have not yet been well characterized. In this study, we used 856 SRAP primer pairs to analyse the 240 individuals from a mapping population, which were derived from a hybrid F₁ generation of ‘Luoxushu 8’ (female) and ‘Zhengshu 20’ (male). Genetic linkage maps of the two parents were constructed. In the female parent (‘Luoxushu 8’), the linkage map consisted of 1391 markers, and the length of the linkage map was estimated to be 10,188.4 cM with an average distance of 7.17 cM between markers. In the male parent (‘Zhengshu 20’), the final linkage map consisted of 1,112 markers, and the estimated length of the map was 9,165.17 cM with an average distance of 8.40 cM between markers. Our results provide a basis for the detailed characterization of sweet potato chromosome sequences and the development of related molecular markers.     

An SSR-based molecular genetic map of cassava

Summary Microsatellites or simple sequence repeats (SSR) are the markers of choice for molecular genetic mapping and marker-assisted selection in many crop species. A microsatellite-based linkage map of cassava was drawn using SSR markers and a F₂ population consisting of 268 individuals. The F₂ population was derived from selfing the genotype K150, an early yielding genotype from an F₁ progeny from a cross between two non-inbred elite cassava varieties, TMS 30572 and CM 2177-2 from IITA and CIAT respectively. A set of 472 SSR markers, previously developed from cassava genomic and cDNA libraries, were screened for polymorphism in K150 and its parents TMS 30572 and CM 2177-2. One hundred and twenty two polymorphic SSR markers were identified and utilized for linkage analysis. The map has 100 markers spanning 1236.7 cM, distributed on 22 linkage groups with an average marker distance of 17.92 cM. Marker density across the genome was uniform. This is the first SSR based linkage map of cassava and represents an important step towards quantitative trait loci mapping and genetic analysis of complex traits in M. esculenta species in national research program and other institutes with minimal laboratory facilities. SSR markers reduce the time and cost of mapping quantitative trait loci (QTL) controlling traits of agronomic interest, and are of potential use for marker-assisted selection (MAS).     

利用3个F_2群体整合高密度栽培种花生遗传连锁图

遗传图谱的构建及整合是开展花生分子育种研究的基础,利用多个作图群体整合遗传图谱是解决图谱标记密度低的有效途径。本研究采用基于锚定SSR标记的作图策略,构建3个F_2群体3张遗传连锁图,利用Join Map 3.0软件整合图谱,获得一张包含20个连锁群、792个位点、总遗传距离为2079.50 c M,标记间平均距离为2.63 c M的整合图谱,各连锁群标记数在20~66个之间,遗传距离在59.10~175.80 c M之间。将3个分离群体中检测到的与荚果及种子大小相关的QTL区段与整合连锁图的标记比较发现,各群体中检测到的位于各染色体上的QTL在整合图谱中都能出现,有些QTL标记区间在整合图谱中存在更多的标记,为今后利用这些标记进行精细定位奠定了基础。     

Density enhancement of a faba bean genetic linkage map (Vicia faba) based on simple sequence repeats markers

Genetic mapping for faba bean lags far behind other major crops. Density enhancement of the faba bean genetic linkage map was carried out by screening 5,325 genomic SSR primers and 2033 expressed sequence tag (EST)‐SSR primers on the parental cultivars '91825' and 'K1563'. Two hundred and fifteen genomic SSR and 133 EST‐SSR primer pairs that detected polymorphisms in the parents were used to screen 129 F₂ individuals. This study added 337 more SSR markers and extended the previous linkage map by 2928.45 cM to a total of 4516.75 cM. The number of SSR markers in the linkage groups varied from 12 to 136 while the length of each linkage group ranged from 129.35 to 1180.21 cM. The average distance between adjacent loci in the enhanced genetic linkage map was 9.71 cM, which is 2.79 cM shorter than the first linkage map of faba bean. The density‐enhanced genetic map of faba bean will be useful for marker‐assisted selection and breeding in this important legume crop.     

The first genetic linkage map of crape myrtle (Lagerstroemia) based on amplification fragment length polymorphisms and simple sequence repeats markers

Lagerstroemia (crape myrtle) are famous ornamental plants with large pyramidal racemes, long flower duration and diverse colours. Genetic maps provide an important genomic resource of basic and applied significance. A genetic linkage map was developed by genotyping 192 F₁ progeny from a cross between L. caudata (female) and L. indica (‘Xiang Xue Yun’) (male) with a combination of amplification fragment length polymorphisms (AFLP) and simple sequence repeats (SSR) markers in a double pseudo‐testcross mapping strategy. A total of 330 polymorphic loci consisting of 284 AFLPs and 46 SSRs showing Mendelian segregation were generated from 383 AFLP primer combinations and 150 SSR primers. The data were analysed using JoinMap 4.0 (evaluation version) to construct the linkage map. The map consisted of 20 linkage groups of 173 loci (160 AFLPs and 13 SSRs) covering 1162.1 cM with a mean distance of 10.69 cM between adjacent markers. The 20 linkage groups contained 2–49 loci and ranged in length from 7.38 to 163.57 cM. This map will serve as a framework for mapping QTLs and provide reference information for future molecular breeding work.     

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.     

利用2个F_2群体整合中国豌豆高密度SSR遗传连锁图谱

豌豆(Pisum sativum L.)是一种重要的食用豆类作物,在全世界范围内广泛种植,既可作为人类食物,也可作为牲畜饲料。用SSR标记构建的遗传连锁图谱在豌豆和其他作物的标记辅助育种中发挥着重要的作用。尽管对豌豆遗传连锁作图的研究已有悠久历史,但公众可获得且可转移的SSR标记以及基于遗传独特的中国豌豆种质的高密度遗传连锁图谱仍然有限。为了获得更多可转移的SSR标记和中国豌豆的高密度遗传连锁图谱,本研究首先从自主开发和文献获取的12,491个全基因组SSR标记中筛选了617个多态性SSR标记,并用于G0003973×G0005527 F_2群体遗传连锁图谱的加密。加密后的图谱全长扩展到5330.6 cM,包含603个SSR标记,标记平均间距离8.8 cM,相比之前的图谱有明显改善。基于上述结果,我们又筛选了119个具有多态性的SSR标记,用于构建大样本W6-22600×W6-15174 F_2群体的遗传连锁图谱,新图谱累积长度为1127.1 cM,包含118个SSR标记,装配在7条连锁群上。最后,将来自以上2个遗传图谱的数据进行整合,得到了一张覆盖范围6592.6 cM的整合图谱,包含668个SSR标记,由509个基因组SSR、134个EST-SSR和25个锚定标记组成,分布在7条连锁群上。这些SSR标记和遗传连锁图谱将为豌豆的遗传研究和标记辅助育种提供有力工具。     

栽培种花生遗传图谱的构建及主茎高和总分枝数QTL分析

栽培种花生是异源四倍体,基因组大,构建花生的分子遗传连锁图谱并对相关性状进行QTL定位研究的工作缓慢。本研究以遗传差异大的亲本组配杂交组合富川大花生×ICG6375构建F2作图群体,采用公开发表的2653对SSR引物,构建了一张含有234个SSR标记、分布于20个连锁群的栽培种花生遗传图谱。该图谱覆盖基因组的长度为1683.43c M,各个连锁群长度在36.11~131.48 c M之间,每个连锁群的标记数在6~15个之间,标记间的平均距离为7.19 c M。结合F3在湖北武汉和阳逻环境下的主茎高和总分枝数鉴定结果,应用Win QTLCart 2.5软件采用复合区间作图法进行了QTL定位和遗传效应分析。共检测到17个与主茎高和总分枝数相关的QTL位点,贡献率在0.10%~10.22%之间,分布于8个连锁群上。综合分析武汉和阳逻环境的鉴定结果,获得重复一致的与主茎高相关的6个QTL,其中q MHA061.1和q MHA062.1位于连锁群LG06上TC1A2~AHGS0153标记区间,贡献率为5.49%~8.95%;q MHA061.2和q MHA062.2位于LG06上AHGS1375~PM377标记区间,贡献率为2.93%~5.83%;q MHA092.2和q MHA091.1位于连锁群LG09上GM2839~EM87标记区间,贡献率为0.53%~9.43%。     

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.     

甘蔗SSR和AFLP分子遗传连锁图谱构建

采用甘蔗商业品种Co419与野生种割手密Y75/1/2杂交,获得269个单株,组成F1群体,用F102/356与商业品种ROC25回交获得266个单株,组成BC1群体。利用筛选的多态性条带丰富的36对SSR引物和12对AFLP引物,对两个群体进行PCR扩增和分子遗传连锁分析,构建甘蔗分子遗传连锁图谱。用F1群体获得630个分离标记,经χ2检测,298个标记为单双剂量标记,占总标记数的47%;用BC1群体获得571个分离标记,有264个标记为单双剂量标记,占总标记数的46%;4个亲本获得单双剂量标记的数量依次为Co41902/356Y75/1/2ROC25。在LOD≥5.0,相邻标记遗传距离≤40cM的条件下,F1群体有134个单双剂量标记被纳入55个连锁群,其中39个连锁群归属8个同源组,16个未列入,总遗传距离为1458.3cM,标记间平均图距为10.9cM;BC1群体有133个单双剂量标记被纳入47个连锁群,其中34个连锁群归属于8个同源组,13个连锁群未列入,总遗传距离为1059.6cM,标记间平均图距为8.0cM。从4个亲本单双剂量标记进入的连锁群数来看,Co419最多,归入34个连锁群,其次为Y75/1/2,归入20个连锁群,第3为02/356和ROC25,归入19个连锁群。研究结果表明,从单双剂量标记比例、形成连锁群数量、总遗传距离来看,F1群体构图质量要优于BC1群体。     

YM型小麦温敏雄性不育系不育基因的QTL定位

YM型小麦温敏雄性不育系的不育基因被定位在1Bs染色体片段上, 但已发现的相邻分子标记与该基因的遗传距离较大, 达10 cM以上。为寻找与该基因连锁更紧密的分子标记, 以YM型温敏雄性不育系ATM3314与恢复系中国春杂交的F₂代200株为作图群体, 从1Bs的22个SSR引物中筛选出5个在亲本和F₂代中分离的SSR引物, 构建了1个包含5个标记的1Bs局部遗传连锁图谱。结合F₂代个体的育性调查, 采用复合区间作图法在YM型温敏雄性不育系的1Bs染色体上检测到不育基因的1个主效QTLrfv1-1和1个微效QTLrfv1-2。rfv1-1位于SSR标记Xgwm18和Xwmc406之间, 与两标记的遗传距离分别为6.0 cM和4.6 cM, LOD值为8.80, 加性效应23.87, 显性效应10.44, 可解释表型变异的23.91%; rfv1-2位于Xwmc406和Xbarc8之间, 与两标记的遗传距离分别为4.0 cM和3.4 cM, LOD值为3.10, 加性效应17.59, 显性效应5.99, 可解释表型变异的7.78%。本研究初步定位了YM型小麦温敏雄性不育系1Bs染色体片段上不育基因的QTL, 为进一步准确定位该基因奠定了基础。     

Comparison and integration of genetic maps generated from F2 and BC1-type mapping populations in perennial ryegrass

Linkage maps of perennial ryegrass were constructed from F₂ and BC₁‐type populations using, predominantly, restriction fragment length polymorphism data based on heterologous probes used in mapping other grass species. The maps identified seven linkage groups, which covered a total of 515 cM (F₂) and 565 cM (BC₁). They were aligned using 38 loci identified in both populations (common loci) and a possible marker order for all mapped loci in either population was identified in an integrated map. The estimated recombination frequencies and map distances between adjacent common loci were compared between the two data sets and regions of heterogeneity identified. Overall, the common markers identified a map distance of 446 cM in the F₂ population and 327 cM in the BC₁ population, reflecting a higher recombination frequency in the former, although the difference was not evenly spread over the seven linkage groups.     

Different recombination frequencies in wheat doubled haploid populations obtained through maize pollination and anther culture

This study compared the meiotic recombination frequency between wheat doubled haploid (DH) populations obtained through two different methods, maize pollination (MP♀) and anther culture (AC♂). The comparison was based on a genetic linkage analysis, performed with DNA markers. Thirty-five polymorphic markers (15 SSR, 15 AFLP, 5 RAPD) were screened in MP♀ and AC♂ doubled haploids populations, derived from the same hybrid genotype (F₁ of ‘Eta’ × ‘Darkhan 15’). Nine linkage groups, comprising 35 loci (the MP♀ lines) and 31 loci (the AC♂ lines), were constructed. The linkage groups in both DH populations showed identical orders of markers, except for one group mapping to chromosome 6B. The MP♀ and AC♂ linkage maps differed significantly in recombination frequencies for corresponding intervals. In total, the AC♂ linkage map (495.5 cM) was 40.5% longer than the MP♀ map (352.8 cM), indicating a significantly higher meiotic recombination rate in pollen mother cells. The enhancement in recombination was visible in five of nine linkage groups, and in 7 intervals between individual loci out of 19 compared. Moreover, for 6 other intervals a lack of linkage was observed in the AC♂ population, as compared to the MP♀ map.     

Construction of a high-density reference linkage map of tea (Camellia sinensis)

A few linkage maps of tea have been constructed using pseudo-testcross theory based on dominant marker systems. However, dominant markers are not suitable as landmark markers across a wide range of materials. Therefore, we developed co-dominant SSR markers from genomic DNA and ESTs and constructed a reference map using these co-dominant markers as landmarks. A population of 54 F₁ clones derived from reciprocal crosses between ‘Sayamakaori’ and ‘Kana-Ck17’ was used for the linkage analysis. Maps of both parents were constructed from the F₁ population that was taken for BC₁ population. The order of most of the dominant markers in the parental maps was consistent. We constructed a core map by merging the linkage data for markers that detected polymorphisms in both parents. The core map contains 15 linkage groups, which corresponds to the basic chromosome number of tea. The total length of the core map is 1218 cM. Here, we present the reference map as a central core map sandwiched between the parental maps for each linkage group; the combined maps contain 441 SSRs, 7 CAPS, 2 STS and 674 RAPDs. This newly constructed linkage map can be used as a basic reference linkage map of tea.     

Molecular marker-based genetic linkage map of a diploid banana population (Musa acuminata Colla)

A well-saturated genetic linkage map is valuable for fundamental and applied genetic research. Genetic linkage maps of two half-sib diploid banana populations were constructed using allele-specific-polymerase chain reactions (AS-PCRs), diversity array technology (DArT), and simple sequence repeat (SSR) markers. Molecular maps were produced for each parent using the pseudo-testcross mapping strategy. The first maternal map (6142-1, 81 individuals) consisted of 231 markers divided as followed: 121 DArT, 106 SSR and 4 AS-PCR markers in 15 linkage groups (LGs) covering 670?cM. The second maternal map (6142-1-S, 58 individuals) contained a total of 152 markers including 71 DArTs, 79 SSRs, and 2 AS-PCRs mapped to 16 LGs that spanned 698?cM. The combined paternal map (139 individuals) comprised 316 markers (196 DArTs, 117 SSRs and 3 AS-PCRs) distributed over 15 LGs with a total map length of 1,004?cM. While distorted segregation of some markers was observed in all maps, this was much more frequent for the male parent. Homology between maps was assessed using common markers. While there was generally good congruity with regard to marker order across maps, incongruity in other cases may reflect chromosomal rearrangement events such as inversions, translocations, or deletions. The new banana map can provide a better understanding of the Musa genome and could be used for the identification of economically important traits and improvement of breeding strategies.     

Development of tomato SSR markers from anchored BAC clones of chromosome 12 and their application for genetic diversity analysis and linkage mapping

In this study, we developed a total of 37 simple sequence repeat (SSR) markers from 11 bacterial artificial chromosome (BAC) clone sequences anchored on chromosome 12 of tomato available at Solanaceae Genomics Network. These SSR markers could group a set of 16 tomato genotypes comprising of Solanum lycopersicum, S. pimpinellifolium, S. habrochaites, and S. pennellii unambiguously according to their known species status. Clear subgroups of genotypes within S. lycopersicum were also observed. A subset of 16 SSR markers representing the 11 BAC clones was used for developing genetic linkage maps of three interspecific F₂ populations produced from the crosses involving a common S. lycopersicum parent (CLN2498E) with S. pennellii (LA1940), S. habrochaites (LA407) and S. pimpinellifolium (LA1579). The length of the genetic linkage maps were 112.5 cM, 109.3 cM and 114.1 cM, respectively. Finally, an integrated genetic linkage map spanning a total length of 118.7 cM was developed. The reported SSR markers are uniformly distributed on chromosome 12 and would be useful for genetic diversity and mapping studies in tomato.     

SSR作为锚定标记构建白菜×芜菁分子遗传图谱

为了将已有的大白菜分子遗传图谱和已发表的A基因组参考图谱对应起来,本研究利用国际上发表的大白菜和甘蓝型油菜A基因组特异SSR标记作为锚定标记,重新构建了白菜×芜菁分子遗传图谱。利用双亲和F1对326个SSR标记进行了筛选,共获得86个多态性分子标记。在此基础上整合了已有的400个标记,最终构建了一张由10个连锁群组成,包含了347个标记的分子连锁图谱,图谱总长度为1008.7cM,标记间的平均图距为2.91cM。此图谱上包含了已在A基因组参考图谱上定位的56个SSR标记,分布于10个连锁群上,从而将各个连锁群与参考图谱的连锁群对应起来。每个连锁群上的标记数在25～58个之间,连锁群长度在60.6～177.0cM范围内,平均图距在1.33～4.92cM之间。该图谱为白菜重要性状的遗传定位奠定了良好基础。     

Construction of a high-density SSR marker-based linkage map of zoysiagrass (<Emphasis Type="Italic">Zoysia japonica</Emphasis> Steud.)

A consensus genetic linkage map with 447 SSR markers was constructed for zoysiagrass (Zoysia japonica Steud.), using 86 F₁ individuals from the cross ‘Muroran 2’ × ‘Tawarayama Kita 1’. The consensus map identified 22 linkage groups and had a total length of 2,009.9 cM, with an average map density of 4.8 cM. When compared with a previous AFLP-SSR linkage map, the SSR markers from each linkage group mapped to similar positions in both maps. Eight pairs of linkage groups from the AFLP-SSR map were joined into eight new groups in the current map. This zoysiagrass consensus map contained 35 SSR markers exhibiting high homology with rice genomic sequences from known chromosomal locations. This allowed synteny to be identified between Zoysiagrass linkage groups 2, 3, 9, 19 and rice chromosomes 3, 12, 2, 7 respectively. These results provide important comparative genomics information and the new map is now available for quantitative trait locus analysis, marker-assisted selection and breeding for important traits in zoysiagrass.     

Construction of a genetic linkage map and QTL analysis of fiber-related traits in upland cotton (Gossypium hirsutum L.)

A genetic linkage map with 70 loci (55 SSR, 12 AFLP and 3 morphological loci) was constructed using 117 F₂ plants obtained from a cross between two upland cotton cultivars Yumian 1 and T586, which have relatively high levels of DNA marker polymorphism and differ remarkably in fiber-related traits. The linkage map comprised of 20 linkage groups, covering 525 cM with an average distance of 7.5 cM between two markers, or approximately 11.8% of the recombination length of the cotton genome. The present genetic linkage map was used to identify and map the quantitative trait loci (QTLs) affecting lint percentage and fiber quality traits in 117 F_2:3 family lines. Sixteen QTLs for lint percentage and fiber quality traits were identified in six linkage groups by multiple interval mapping: four QTLs for lint percentage, two QTLs for fiber 2.5% span length, three QTLs for fiber length uniformity, three QTLs for fiber strength, two QTLs for fiber elongation and two QTLs for micronaire reading. The QTL controlling fiber-related traits were mainly additive, and meanwhile including dominant and overdominant. Several QTLs affecting different fiber-related traits were detected within the same chromosome region, suggesting that genes controlling fiber traits may be linked or the result of pleiotropy.     

Male and female genetic linkage map of hops, Humulus lupulus

A male and female linkage map of hop has been constructed using 224 DNA polymorphisms (106 amplified fragment length polymorphisms (AFLPs), three random amplified polymorphic DNAs (RAPDs), one RAPD‐sequence‐tagged‐site (STS), and three microsatellite (STSs) segregating in an F₁ population of the English cultivar ‘Wye Target’‐the German male breeding line ‘85/54/15’. Linkage between these loci was estimated using JOINMAP Version 2.0. The final map for the female parent consisted of 110 loci assigned to eight linkage groups covering a distance of 346.7 cM. For the male map, 57 loci could be mapped on nine linkage groups spanning over 227.4 cM. One of these male linkage groups (Gr09‐M) presumably represents the Y chromosome, since all markers assigned (10 AFLPs, three RAPDs and one STS) were closely linked to the male sex (M). Because of their sex‐specific segregation, 10 doubly heterozygous AFLPs spanning a distance of 18.7 cM could be identified as markers describing the X chromosome, which is part of the male and female map. Three STMSs, which had already proved useful in hop genotyping, could be integrated as codominant locus‐specific markers and thus allowed to produce reliable allelic bridges between the female and male counterparts.