Construction of AFLP-based genetic linkage maps for the Chinese shrimp Fenneropaeneus chinensis

Fenneropaeneus chinensis is an important species in marine fishery resources and aquaculture in China. A genetic linkage map is essential for improving the efficiency of its breeding by marker-assisted selection and identifying commercially important genes. Linkage maps of F. chinensis were constructed with an F2 mapping population （110 progenies） using amplified fragment length polymorphic （AFLP） marker in this study. Fifty-five AFLP primer combinations produced 532 AFLP markers fitting for map strategy in mapping family. The markers with 3：1 segregating ratios were analyzed using F2 intercross model for the common linkage map, while the markers with 1：1 ratio were analyzed using the pseudo-testcross strategy. The maps of male, female and common were constructed. The female map included 103 markers that formed 28 linkage groups, covering a total length of 1090 cM. All markers were linked with the linkage groups. Segregation distortion was observed for 6 of 103 markers in the female map. The average distance between markers was 14.53 cM and ranged from 4.4 to 24.8 cM. The male map included 144 markers that formed 35 linkage groups. Ten markers remained unlinked in male map. Segregation distortion was observed for 7 of 144 markers in the male map. The total distance of male map covered 1617 cM. The average distance between markers was 16.36 cM. The male map was 32.6% longer than the female map, which may reflect sex-specific recombination rates in Chinese shrimp. The common map was composed of 216 markers, including in 44 linkage groups covering a total distance of 1772.1 cM. Two markers remained unlinked. No distorted markers of 216 markers were shown in the common map. The distance between markers was 10.42 cM. An average estimated genome size for the Chinese shrimp was 2420 cM, which was consistent with the relative size of the Penaeid genome. The distribution of AFLP markers was relatively even in chromosomes of Chinese shrimp maps. The linkage analysis presented in this work provided some insight     

Construction of a genetic linkage map for cotton based on SRAP

DNA markers have been widely used in construction of molecular genetic linkage maps in plants. The first genetic linkage map of cotton was constructed by Reinish in 1994 using RFLP (restriction fragment length polymorphism)[1], which included 705 polymorphic loci on 41 linkage groups with a total length of 4675 cM. Afterwards, several genetic linkage maps were constructed[2—7], but no map is comparable to this one in marker density. A high-density genetic linkage map could be applied effec…     

Construction of a genetic map with SRAP markers and localization of the gene responsible for the first-flower-node trait in cucumber (Cucumis sativus L.)

Cucumber (Cucumis sativus L.) is an annualclimber originated from the tropic rain forest area inthe south of Himalayas, which belongs to the Cucur bitaceae family. Cucumber is one of the importantvegetables in the world, and its planting area is sec ond only to that of tomato[1]. However genetics re search on cucumber obviously drops behind that of thelatter. The classic genetic map of cucumber, with sixlinkage groups, is composed of more than 100 genesfor color, morphology, and dise…     

Localization of genes for lateral branch and female sex expression and construction of a molecular linkage map in cucumber (Cucumis sativus L. ) with RAPD markers

A cucumber ( Cucumis sativus L. ) molecular linkage map, including 79 random-amplified polymorphic DNAs (RAPD)and two genes , lb for lateral branch and f for female sex expression, is constructed from a cross between a line, S52, with weak lateral growing ability and staminate from Dabieshan Mountains area in China and another line, S06, with strong lateral growing ability and gynoecious from Europe. The map contains nine linkage groups and spans 1110.0 cM with an average distance of 13.7 cM between loci. The lb locus is located in a longer linkage group LG-2 and flanked by two markers, OP-Q5-1 and OP-M-2-2, at 9.3 cM and 15.9 cM, respectively. In the meantime, the RAPD loci, OP-Q5-2 and BC151, in a short linkage group were found to flank f at 13.7 cM and 13.4 cM,respectively. The construction of RAPD map has paved a way for further study of the genes for lateral branch, female sex expression and other agronomic traits in cucumber.     

Fine mapping of the <Emphasis Type="Italic">Ht2 (Helminthosporium turcicum</Emphasis> resistance 2) gene in maize

Fine mapping of Helminthosporium turcicum resistance gene Ht2 is extremely valuable for map-based cloning of the Ht2 gene,gaining a better knowledge of the distribution of resistance genes in maize genome and marker-assisted selection in maize breeding.An F2 mapping population was developed from a cross between a resistant inbred line 77Ht2 and a susceptible inbred line Huobai.With the aid of RFLP marker analyses,the Ht2 gene was mapped between the RFLP markers UMC89 and BNL2.369on chromosome 8,with a genetic distance of 0.9cM to BNL2.369.There was a linkage between SSR markers UMC1202,BNLG1152,UMC1149 and the Ht2 gene by SSR assay,Among the SSR markers,the genetic distance between UMC1149 and the Ht2 gene was 7.2cM,By bulked segregant analysis 7 RAPD-amplified products which were probably linked to the Ht2 gene were selected after screening 450 RAPD primers and converted the single-copy ones into SCAR markers.Linkage analysis showed that the genetic distance between the SCAR marker SD-06633 and the Ht2 gene was 0.4cM.From these results,a part of linkage map around the Ht2 gene was constructed.     

Reconstruction of linkage maps in the distorted segregation populations of backcross, doubled haploid and recombinant inbred lines

Non-Mendelian segregation of markers, known as distorted segregation, is a common biological phenomenon. Although segregation distortion affects the estimation of map distances and the results of quantitative trait loci (QTL) mapping, the effects of distorted markers are often ignored in the construction of linkage maps and in QTL mapping. Recently, we have developed a multipoint method via a Hidden Markov chain method to reconstruct linkage maps in an F2 population that corrects for bias of map distances between distorted markers. In this article, the method is extended to cover backcross, doubled haploid and recombinant inbred line (RIL) populations. The results from simulated experiments show that: (1) the degree that two linked segregation distortion loci (SDL) affect the estimation of map distances increases as SDL heritability and interval length between adjacent markers increase, whereas sample size has little effect on the bias; (2) two linked SDL result in the underesti- mation of linkage distances for most cases, overestimation for an additive model with opposite additive effects, and unbiased estimation for an epistatic model with negative additive-by-additive effects; (3) the proposed method can obtain the unbiased estimation of linkage distance. This new method was applied to a rice RIL population with severely distorted segregation to reconstruct the linkage maps, and a bootstrap method was used to obtain 95% confidence intervals of map distances. The results from real data analysis further demonstrate the utility of our method, which provides a foundation for the inheritance analysis of quantitative and viability traits.     

Construction of a genetic map and localization of QTLs for yield traits in tomato by SSR markers

Using an F2 population derived from the hybrid of Lycopersicon esculentum Mill. ‘XF 98-7’× Lycopersicon pimpinellifolium LA2184, a SSR genetic linkage map of tomato is constructed. The map contains 112 markers and spans 808.4 cM with an average distance of 7.22 cM between loci. Two quantitative trait loci (QTLs) for first flower node on chromosomes 5 and 11, two QTLs for number of flowers per truss on chromosomes 2 and 5, and five QTLs for fruit weight on chromosomes 1, 2, 3, 9 and 12 are identified.     

利用DNA微卫星标记定位水稻的抗稻瘟病基因

利用回交育种中产生的回交群体结合前人的研究结果构建了Pil基因区域的局部分子标记连锁图，通过BC1F2家系的接种结果判断其基因型，将Pil定位在RFLP标记RZ536与SSR标记RM144之间，图距分别为9．7、6．8cm，从而建立了一套完整的以PCR为基础的分子标记辅助选择体系。     

春小麦重组自交系及多种SSR标记构建遗传图谱

 以春小麦重组自交系(RIL)宁春4 号×宁春27 号为作图群体,利用SSR 标记构建小麦遗传连锁图谱。结果表明,用1001对SSR 引物选出亲本间表现多态性的引物307 对,多态性频率为30.7%。利用307 对多态性引物对RIL 群体进行分析,共检测到266 个多态性标记位点。通过χ²检测(P<0.05),有147 个SSR 标记表现为偏分离,偏分离率为55.3%,129 个偏向母本宁春4号,其偏分离位点主要分布在B 和D 基因组上。用Mapmaker 3.0 和Mapdraw 2.1 软件将266 个SSR 位点绘制在小麦遗传连锁图上,该图谱覆盖小麦基因组全长2187.79 cM,标记间的平均遗传距离为8.22 cM。     

Verification and fine-mapping of QTLs conferring days to flowering in soybean using residual heterozygous lines

The results of QTL mapping based on a primary mapping population should be further verified and refined for its real utilization in marker-assisted selection or map-based cloning.The primary mapping population contains 114 BC1F1 plants of the backcross between Essex (maturity group V,MG V) as the donor parent and ZDD2315 (MG II) as the recurrent parent.In this study,a genetic linkage map with 250 SSR markers spanning a total length of 2963.5 cM on 25 linkage groups (LG) was constructed using software MAPMAK...     

Chicken QTL mapping by multiplex PCR

To facilitate rapid determination of the chromosomal location of quantitative trait loci' the current approaches to gene mapping are improved using a multiplex PCR technique. The high-throughput linkage analysis method described here allows selection of 178 from 328 microsatellite markers through the multiplex PCR method combined with the semi-automatic fluorescence-labeled DNA analysis technology. Those polymorphism markers are distributed on 23 autosomes and one sex chromosome (chromosome Z). covering 3080cM genetic distance. The average marker density is 18cM. dispersed into 30 different sets. These selected polymorphism microsatellite markers segregate with the family members, following the Mendel's heritage laws, and are very useful for chicken linkage map analysis as well as for the research on some important economic quantitative characters of chicken.     

基于SLAF-seq玉米高密度遗传连锁图谱构建与株型性状QTL定位

 株型直接决定生物产量、种植密度与籽粒产量,利用玉米高密度遗传连锁图谱解析株型相关性状的遗传机制,对选育理想株型玉米新品种具有重要意义。本研究利用SLAF-seq技术,依据玉米黄早四参考基因组信息,对昌7-2与PHB1M及其138个F_2：3家系高通量测序,开发高密度SNP的遗传图谱,并进行株型相关性状QTL定位。研究结果构建了一张总图距为1354.81 cM,标记间的平均距离为0.32 cM,标记数为4220个SLAF标签（7876个SNP）的高密度遗传图谱。在E1与E2两种密度下,对株高、穗位、叶片数、节间数、平均节间长进行QTL定位,共检测到10个QTL位点,其中,有7个PVE超过了10%。叶片数、穗位、节间数为主效QTL,ADD为负值,叶片数与节间数的减效等位基因来源于PHB1M,穗位的减效等位基因来源于昌7-2。叶片数与节间数在2个密度下均定位在8号染色体上,说明二者之间有着共同的遗传机制。与QTL关联的SLAF标记共有61个,其中,SLFA7305498和SLFA6717271为qLC-2-LG8与qIC-2-LG8共有标记。该研究将为玉米株型相关性状的标记辅助选择提供支撑。     

Fine mapping of the dominant glandless Gene Gl2^e in Sea-island cotton （Gossypium barbadense L.）

Gle2 is a mutant gene that controls glandless trait in cotton plants and seeds. It is an important gene resource to gossypol-free cottonseed breeding. The objective of this research was to develop SSR markers tightly linked with Gle2 by using the F2 segregating population containing 1599 plants derived from the cross of G. hirsutum genetic standard line TM-1 and G. barbadense glandless mutant line Hai-1. Genetic analysis suggested that the Gle2 was an incomplete dominant gene. Based on the backbone of genetic linkage map from G. hirsutum × G. barbadense BC1 published by our laboratory,Gle2 was lo-cated between CIR362 and NAU2251b,NAU3860b,STV033,with a genetic distance 9.27 and 0.96 cM,respectively. This result is useful for cloning Gle2 gene by map-based cloning method.     

Construction of a genetic map and location of quantitative trait loci for dwarf trait in maize by RFLP markers

Using F₂ population derived from the cross of tall inbred 7922 by dwarf inbred 5003, an RFLP linkage map of maize has been constructed, on which 85 markers are distributed among 10 linkage groups and span maize genome about 1827.8 cM with an average distance (24.4 cM) between markers. 106 F_2:3 lines of the population were grown in a 10 × 11 simple rectangular lattice design of one-raw plots with two replications and evaluated for plant height (PH). With interval mapping procedure, 5 QTLs controlling plant height have been identified and their genetic effects and gene action determined. 2 major QTLs with opposite effect have been discovered. One for increasing plant height isph1 which is located at chromosome 2 and accounts for 51.8% of the total phenotypic variation; the other for decreasing plant height isph3 which is located at chromosome 5 and accounts for 38.6% of the total phenotypic variation. The chromosomal location ofph3 might be the same as or close to the position ofbv1, a dwarf mutant of maize.     

Fine mapping of an incomplete recessive gene for leaf rolling in rice （Oryza sativa L.）

Genetic analysis and fine mapping of genes controlling leaf rolling were conducted using two backcrossed generations （BC4F2, BC4F3） derived from a cross between QMX, a non-rolled leaf cultivar as a recurrent parent, and JZB, a rolled leaf NIL of ZB as a donor parent. Results indicated that leaf rolling was mainly controlled by an incompletely recessive major gene, namely rl（t）, and at the same time, affected by quantitative trait loci （QTLs） and/or the environment. A genetic linkage map was constructed using MAPMAKER/EXP3.0 with eight polymorphic markers on chromosome 2, which were screened by BAS method from 500 SSR markers and 15 newly developed insertion/deletion （InDel） markers. The position of rl（t） was estimated with composite interval mapping （CIM） method using WinQTLcart2.5. Gene rl（t） was mapped between markers InDel 112 and RM3763, and 1.0 cM away from InDel 112 using 241 plants in BC4F2 population. To fine map r（t）, one BC4F3 line with 855 plants was generated from one semi-rolled leaf plant in BC4F2. Four new polymorphic InDel markers were developed, including InDel 112.6 and InDel 113 located between markers InDe1112 and RM3763. Based on the information of recombination offered by 191 rolled leaf plants and 185 non-rolled leaf plants from the BC4F3 line ,we mapped r（t） to a 137-kb region between markers InDel 112.6 and InDel 113. Homologous gene analysis suggested that r（t）was probably related to the process of leaf development regulated by microRNA.     

A genome scan for quantitative trait loci affecting grain yield and its components of maize both in single-and two-locus levels

Maize is one of the most important cereal crops in the world. The hybrid yield advantage is responsible for about 10 percent of the total global maize production of 550 Mt[1]. It is exigent to study the yield traits so as to improve the hybrids per se in …     

Comparison of human genetic and sequence-based physical maps

Recombination is the exchange of information between two homologous chromosomes during meiosis. The rate of recombination per nucleotide, which profoundly affects the evolution of chromosomal segments, is calculated by comparing genetic and physical maps. Human physical maps have been constructed using cytogenetics, overlapping DNA clones and radiation hybrids; but the ultimate and by far the most accurate physical map is the actual nucleotide sequence. The completion of the draft human genomic sequence provides us with the best opportunity yet to compare the genetic and physical maps. Here we describe our estimates of female, male and sex-average recombination rates for about 60% of the genome. Recombination rates varied greatly along each chromosome, from 0 to at least 9 centiMorgans per megabase (cM Mb(-1)). Among several sequence and marker parameters tested, only relative marker position along the metacentric chromosomes in males correlated strongly with recombination rate. We identified several chromosomal regions up to 6 Mb in length with particularly low (deserts) or high (jungles) recombination rates. Linkage disequilibrium was much more common and extended for greater distances in the deserts than in the jungles.     

石刁柏雄性连锁的AFLP及SCAR标记的建立

目的:对石刁柏雌雄株基因组差异进行分析,筛选雄性或雌性连锁的分子标记.方法:利用限制性片段长度多态性技术,设计了多个引物组合,分别对石刁柏雌雄株基因组进行扩增.结果:在使用的72个引物组合中,引物组合E-AAG+M-CAT从雄性基因组中扩增出了一个雄性连锁的标记(MLDA555),该序列长度为555bp,AT含量为59%.Blast检索未发现相似序列.根据该片段序列设计的引物将该标记转化为雄性连锁的大小为523bp的稳定的SCAR标记,经过不同基因型雄性个体的验证证明该标记广泛存在于不同基因型石刁柏雄性个体中.结论:通过AFLP扩增筛选得到了石刁柏雄性连锁的AFLP和SCAR标记,为石刁柏性别决定机制的理解及石刁柏的分子标记辅助育种提供理论资料和技术支持.     

Characterization and mapping of a white panicle mutant gene in rice

A spontaneous white panicle mutant was found from the F6 progenies of an indicajaponica cross.The mutant exhibits white stripes on its basal leaves while the panicles,rachis and pedicel are milky white colored at flowering stage.Genetic analysis in an F2 population from the cross of Zhi7/white panicle mutant indicates that the white panicle phenotype is controlled by a single recessive nuclear gene,tentatively termed as wp(t).Using microsatellite markers,the wp(t) gene was anchored between the markers of SSR101 and SSR63.9 with a map distance of 2.3 and 0.8cM,respectively,and co-segregated with the marker of SSR17 on rice chromosome 1.     

Fine-scale recombination rate differences between sexes, populations and individuals

Meiotic recombinations contribute to genetic diversity by yielding new combinations of alleles. Recently, high-resolution recombination maps were inferred from high-density single-nucleotide polymorphism (SNP) data using linkage disequilibrium (LD) patterns that capture historical recombination events. The use of these maps has been demonstrated by the identification of recombination hotspots and associated motifs, and the discovery that the PRDM9 gene affects the proportion of recombinations occurring at hotspots. However, these maps provide no information about individual or sex differences. Moreover, locus-specific demographic factors like natural selection can bias LD-based estimates of recombination rate. Existing genetic maps based on family data avoid these shortcomings, but their resolution is limited by relatively few meioses and a low density of markers. Here we used genome-wide SNP data from 15,257 parent-offspring pairs to construct the first recombination maps based on directly observed recombinations with a resolution that is effective down to 10 kilobases (kb). Comparing male and female maps reveals that about 15% of hotspots in one sex are specific to that sex. Although male recombinations result in more shuffling of exons within genes, female recombinations generate more new combinations of nearby genes. We discover novel associations between recombination characteristics of individuals and variants in the PRDM9 gene and we identify new recombination hotspots. Comparisons of our maps with two LD-based maps inferred from data of HapMap populations of Utah residents with ancestry from northern and western Europe (CEU) and Yoruba in Ibadan, Nigeria (YRI) reveal population differences previously masked by noise and map differences at regions previously described as targets of natural selection.