QTL mapping for berry shape based on a high-density genetic map constructed by whole-genome resequencing in grape

Grape berry shape is an important agricultural trait.Clarifying its genetic basis is significant for cultivating grape varieties that meet market demands.However,the current study by forward genetics has not achieved in-depth results.Here,a high-density map was constructed to identify quantitative trait loci(QTLs)for berry shape.A total of 358709 polymorphic SNPs were obtained using whole-genome resequencing(WGS)based on 208 F2 individuals derived from round grape‘E42-6’and oblong grape‘Rizamat’.The 1635.65 cM high-density map was divided into 19 linkage groups with an average distance of 0.37 cM.Using this map,three significant QTLs for fruit shape index(ShI:ratio of berry length to berry width)identified over three years were mapped onto LG4 and LG5,including one stable QTL on Chr5 with the genomic region of 0.47–1.94 Mb.Combining with gene annotation and expression patterns based on RNA-seq data from two contrasting F2 individuals with round and oblong berry(their average ShI was 1.89 and 1.10,respectively)at four developmental stages,four candidate genes were selected from the above QTLs.They were mainly involved in DNA replication,cell wall modification,and phytohormone biosynthesis.Further analysis of RNA-seq data revealed that several important phytohormone synthesis and metabolic pathways were enriched based on differentially expressed genes(DEGs),which was consistent with the results of QTL mapping for genes related to plant hormone biosynthesis in the F2 population.Furthermore,a comparison of plant hormone content showed that there were significant differences in IAA and tZ content between the two contrasting F2 individuals at different developmental stages.Our findings provide molecular insights into the genetic variation in grape berry shape.Stable QTLs and their tightly linked markers offer the possibility of marker-assisted selection to accelerate berry shape breeding.

Breeding design in wheat by combining the QTL information in a GWAS panel with a general genetic map and computer simulation

A large amount of genome-wide association study(GWAS)panels together with quantitative-trait locus(QTL)information associated with breeding-targeted traits have been described in wheat(Triticum aestivum L.).However,the application of mapping results from a GWAS panel to conventional wheat breeding remains a challenge.In this study,we first report a general genetic map which was constructed from 44 published linkage maps.It permits the estimation of genetic distances between any two genetic loci with physical map positions,thereby unifying the linkage relationships between QTL,genes,and genomic markers from multiple genetic populations.Second,we describe QTL mapping in a wheat GWAS panel of 688 accessions,identifying 77 QTL associated with 12 yield and grain-quality traits.Because these QTL have known physical map positions,they could be mapped onto the general map.Finally,we present a design approach to wheat breeding by using known QTL information and computer simulation.Potential crosses between parents in the GWAS panel may be evaluated by the relative frequency of the target genotype,trait correlations in simulated progeny populations,and genetic gain of selected progenies.It is possible to simultaneously improve yield and grain quality by suitable parental selection,progeny population size,and progeny selection scheme.Applying the design approach will allow identifying the most promising crosses and selection schemes in advance of the field experiment,increasing predictability and efficiency in wheat breeding.

Construction of SNP genetic maps based on targeted next-generation sequencing and QTL mapping of vital agronomic traits in faba bean(Vicia faba L.)

Owing to the limitation of a large genome size(~13 Gb),the genetic and gene mapping studies on faba bean(Vicia faba L.)are lagging far behind those for other legumes.In this study,we selected three purified faba bean lines(Yundou 8137,H0003712,and H000572)as parents and constructed two F2 populations.These two F2 populations,namely 167 F2 plants in Pop1(Yundou 8137×H0003712)and 204 F2 plants in Pop2(H000572×Yundou 8137),were genotyped using a targeted next-generation sequencing(TNGS)genotyping platform,and two high-density single nucleotide polymorphisms(SNP)genetic linkage maps of faba bean were constructed.The map constructed from Pop1 contained 5103 SNPs with a length of 1333.31 cM and an average marker density of 0.26 cM.The map constructed from Pop2 contained 1904 SNPs with a greater length of 1610.61 cM.In these two F2 populations,QTL mapping identified 98 QTLs for 14 agronomic traits related to the flowers,pods,plant types and grains.The two maps were then merged into an integrated genetic linkage map containing 6895 SNPs,with a length of 3324.48 cM.These results not only lay the foundation for fine mapping and map-based cloning of related genes,but can also accelerate the molecular marker-assisted breeding of faba bean.

Construction of high-density SNP genetic maps and QTL mapping for dwarf-related traits in Litchi chinensis Sonn

Litchi chinensis Sonn is widely cultivated in subtropical regions and has an important economic value.A high-density genetic map is a valuable tool for mapping quantitative trait loci(QTL)and marker-assisted breeding programs.In this study,a single nucleotide polymorphism(SNP)-based high-density linkage map was constructed by a genotyping-by-sequencing(GBS)protocol using an F1 population of 178 progenies between two commercial litchi cultivars,‘Ziniangxi’(dwarf)and‘Feizixiao’(vigorous).The genetic map consisted of 3027 SNP markers with a total length of 1711.97 cM in 15 linkage groups(LGs)and an average marker distance of 0.57 cM.Based on this high-density linkage map and three years of phenotyping,a total of 37 QTLs were detected for eight dwarf-related traits,including length of new branch(LNB),diameter of new branch(DNB),length of common petiole(LCP),diameter of common petiole(DCP),length of internode(LI),length of single leaf(LSL),width of single leaf(WSL),and plant height(PH).These QTLs could explain 8.0 to 14.7%(mean=9.7%)of the phenotypic variation.Among them,several QTL clusters were observed,particularly on LG04 and LG11,which might show enrichment for genes regulating the dwarf-related traits in litchi.There were 126 candidate genes identified within the QTL regions,55 of which are differentially expressed genes by RNA-seq analysis between‘Ziniangxi’and‘Feizixiao’.These DEGs were found to participate in the regulation of cell development,material transportation,signal transduction,and plant morphogenesis,so they might play important roles in regulating plant dwarf-related traits.The high-density genetic map and QTLs identification related to dwarf traits can provide a valuable genetic resource and a basis for marker-assisted selection and genomic studies of litchi.

Construction of the first high-density genetic map for growth related QTL analysis in Ancherythroculter nigrocauda

Ancherythroculter nigrocauda is a fish endemic to the upper areas of the Changjiang(Yangtze)River in China.Quantitative trait locus(QTL)mapping is a powerful tool to identify potential genes affecting traits of economic importance in domestic animals.In this study,a high-density genetic map was constructed with 5901 single nucleotide polymorphism(SNP)makers by sequencing 92 individual fish from a F1 family using the specific-locus amplified fragment sequencing approach.Initially,48 QTLs for total length,body length,body height,and body weight were identified according to the high density of the genetic map with 24 LGs,a total length of 3839.4 cM,and marker spacing of about 0.82 cM.These QTLs explained 27.1%-49.9%of phenotypic variance.The results of this study suggest that major QTLs are responsible for the growth of A.nigrocauda,and these are potentially useful in comparative genomics research,genome assembly,and marker-assisted breeding programs for this species.

Advances and prospects of genetic mapping of Verticillium wilt resistance in cotton

Verticillium wilt is one of the most important diseases affecting cotton production in China.The fungus,Verticillium dahliae,has a wide host range and a high degree of genetic variability.No resistance resources have been found in the available planting resources,thus presenting difficulties and challenges for our study.The long-term production practice shows that selection of disease-resistant varieties is the most economical and effective measure to control Verticillium wilt of cotton to reduce the yield loss and quality decline of cotton.In this paper,we summarized the genetic mapping population,the analysis method of genetic localization,the discovery,mining and cloning of disease-resistant quantitative trait loci/markers,and the analysis of their genetic functions,so as to provide information for the molecular breeding approach of disease-resistant cotton.

High-density genetic mapping identified a major locus for environmental sex expression in pumpkin(Cucurbita moschata Duch.)

Long-day length and high temperature inhibit sex expression in pumpkin(Cucurbita moschata Duch.),and therefore directly impact the production potential.In this study,female flowering patterns in photoperiod-insensitive(PPIS)and photoperiod-sensitive(PPS)germplasms differed significantly in a moderately long day and high temperature environment.However,both germplasms exhibited a similar response in short day with either low temperature or high temperature environment.Photoperiod sensitivity led to this difference in sex expression between the germplasms.For the traits of 1st female flowering node(FFFN)and number of female flowers(NFF),high-density linkage map construction and quantitative trait locus(QTL)mapping were performed using SLAF-seq technology and 162 F_(2) individuals generated from PPIS and PPS.In total,4655 SLAFs were selected and mapped on 20 linkage groups(LGs).The total map length was 2502.01 cM with an average interval distance of 0.75 cM.Major QTLs for both FFFN and NFF were detected on LG6 with intervals of 7.89 and 17.67 cM and PVE values of 30.5%and 22.9%,respectively.Further analyses of the major locus for FFFN revealed 73 protein-coding genes.Among them,4 were related to sex expression,photoperiod flowering,and hormone response.An InDel(insertion and deletion)marker partially correlated with FFFN of the F_(2) population was also developed.Our study identified the QTL for the sex expression response to environmental factors using the high-density linkage map.The identified candidate genes and markers will provide useful information about the molecular interaction between the environment and sex expression and for marker-assisted selection of pumpkin environment-insensitive resources.

Construction of a high-density adzuki bean genetic map and evaluation of its utility based on a QTL analysis of seed size

Adzuki bean(Vigna angularis(Willd.)Ohwi&Ohashi)is an annual cultivated leguminous crop commonly grown in Asia and consumed worldwide.However,there has been limited research regarding adzuki bean genetics,which has prevented the efficient application of genes during breeding.In the present study,we constructed a high-density genetic map based on whole genome re-sequencing technology and validated its utility by mining QTLs related to seed size.Moreover,we analyzed the sequences flanking insertions/deletions(In Dels)to develop a set of PCR-based markers useful for characterizing adzuki bean genetics.A total of 2904 markers were mapped to 11 linkage groups(LGs).The total length of the map was 1365.0 cM,with an average distance between markers of 0.47 cM.Among the LGs,the number of markers ranged from 208(LG7)to 397(LG1)and the total distance ranged from 97.4 cM(LG9)to 155.6 cM(LG1).Twelve QTLs related to seed size were identified using the constructed map.The two major QTLs in LG2 and LG9 explained 22.1 and 18.8%of the total phenotypic variation,respectively.Ten minor QTLs in LG4,LG5 and LG6 explained 3.0–10.4%of the total phenotypic variation.A total of 9718 primer pairs were designed based on the sequences flanking In Dels.Among the 200 selected primer pairs,75 revealed polymorphisms in 24 adzuki bean germplasms.The genetic map constructed in this study will be useful for screening genes related to other traits.Furthermore,the QTL analysis of seed size and the novel markers described herein may be relevant for future molecular investigations of adzuki bean and will be useful for exploiting the mechanisms underlying legume seed development.

Genetic map construction and functional characterization of genes within the segregation distortion regions(SDRs)in the F_(2:3) populations derived from wild cotton species of the D genome

Background:Segregation distortion(SD)is a common phenomenon among stable or segregating populations,and the principle behind it still puzzles many researchers.The F2:3 progenies developed from the wild cotton species of the D genomes were used to investigate the possible plant transcription factors within the segregation distortion regions(SDRs).A consensus map was developed between two maps from the four D genomes,map A derived from F2:3 progenies of Gossypium klotzschianum and G.davidsonii while Map B from G.thurberi and G.trilobum F2:3 generations.In each map,188 individual plants were used.Results:The consensus linkage map had 1492 markers across the 13 linkage groups with a map size of 1467.445 cM and an average marker distance of 1.0370 cM.Chromosome D502 had the highest percentage of SD with 58.6%,followed by Chromosome D507 with 47.9%.Six thousand and thirty-eight genes were mined within the SDRs on chromosome D502 and D507 of the consensus map.Within chromosome D502 and D507,2308 and 3730 genes were mined,respectively,and were found to belong to 1117 gourp out of which 622 groups were common across the two chromosomes.Moreover,genes within the top 9 groups related to plant resistance genes(R genes),whereas 188 genes encoding protein kinase domain(PF00069)comprised the largest group.Further analysis of the dominant gene group revealed that 287 miRNAs were found to target various genes,such as the gra-miR398,gramiR5207,miR164a,miR164b,miR164c among others,which have been found to target top-ranked stress-responsive transcription factors such as NAC genes.Moreover,some of the stress-responsive cis-regulatory elements were also detected.Furthermore,RNA profiling of the genes from the dominant family showed that higher numbers of genes were highly upregulated under salt and osmotic stress conditions,and also they were highly expressed at different stages of fiber development.Conclusion:The results indicated the critical role of the SDRs in the evolution of the key regulatory genes in plants.

Genetic Map of Cotton with Molecular Markers

Cotton (Gossypium spp.) is the most important natural fiber in the world, and its seeds are also used as a food source. Breeding cotton for traits of interest, such as production and processing of fibers, will ensure that this natural product is as competitive as renewable synthetic fibers derived from petroleum. Thus, the mapping of the cotton genome for traits of interest may be the basis for its subsequent use in breeding programs. This work consists of a literature review, with the aim of bringing together works from different research groups working with the mapping of the cotton genome with molecular markers.

High density genetic map and quantitative trait loci(QTLs)associated with petal number and flower diameter identified in tetraploid rose

Rose is one of the most important ornamental and economic plants in the world.Modern rose cultivars are primarily tetraploid,and during meiosis,they may exhibit double reduction or preferential chromosome pairing.Therefore,the construction of a high density genetic map of tetraploid rose is both challenging and instructive.In this study,a tetraploid rose population was used to conduct a genetic analysis using genome sequencing.A total of 17382 single nucleotide polymorphism(SNP)markers were selected from 2308042 detected SNPs.Combined with 440 previously developed simple sequence repeats(SSR)and amplified fragment length polymorphism(AFLP)markers,a marker dosage of 6885 high quality markers was successfully assigned by GATK software in the tetraploid model.These markers were used in the construction of a high density genetic map,containing the expected seven linkage groups with 6842 markers,a total map length of 1158.9 c M,and an average inter-marker distance of 0.18 c M.Quantitative trait locus(QTL)analysis was subsequently performed to characterize the genetic architecture of petal number and flower diameter.One major QTL(qpnum-3-1)was detected for petal number in three consecutive years,which explained 20.18–22.11%of the variation in petal number.Four QTLs were detected for flower diameter;the main locus,qfdia-2-2,was identified in two consecutive years.Our results will benefit the molecular marker-assisted breeding of modern rose cultivars.In addition,this study provides a guide for the genetic and QTL analysis of autotetraploid plants using sequencing-based genotyping methods.

Construction of a Genetic Linkage Map of Mango Based on SRAP, AFLP and ISSR Markers

A hybrid F_1 population was obtained by crossing between mango cultivars‘Jinhuang'and‘Guifei'. Among the hybrid population,98 F_1 plants were selected as the mapping population. The molecular genetic map of mango was constructed by linkage analysis by SRAP,AFLP and ISSR markers using Joinmap 4.0 software. The genetic linkage map consisted of 33 linkage groups,with a total genetic distance of 1561. 1 cM. Moreover,the genetic linkage map involved 245 polymorphic markers,including 149 SRAP markers,90 AFLP markers and 6 ISSR markers,with an average genetic distance of 6. 37 cM. This study laid a foundation for further investigation of the important agronomic traits of mango.

MG2C:a user-friendly online tool for drawing genetic maps

Genetic map is a linear arrangement of the relative positions of sites in the chromosome or genome based on the recombination frequency between genetic markers.It is the important basis for genetic analysis.Several kinds of software have been designed for genetic mapping,but all these tools require users to write or edit code,making it time-costing and difficult for researchers without programming skills to handle with.Here,MG2C,a new online tool was designed,based on PERL and SVG languages.Users can get a standard genetic map,only by providing the location of genes(or quantitative trait loci)and the length of the chromosome,without writing additional code.The operation interface of MG2C contains three sections:data input,data output and parameters.There are 33 attribute parameters in MG2C,which are further divided into 8 modules.Values of the parameters can be changed according to the users’requirements.The information submitted by users will be transformed into the genetic map in SVG file,which can be further modified by other image processing tools.MG2C is a user-friendly and time-saving online tool for drawing genetic maps,especially for those without programming skills.The tool has been running smoothly since 2015,and updated to version 2.1.It significantly lowers the technical barriers for the users,and provides great convenience for the researchers.

Construction of a high-density genetic linkage map and identification of flowering-related QTL in erect milkvetch(Astragalus adsurgens)

Erect milkvetch(Astragalus adsurgens)is a perennial legume forage crop with economic and ecological value in livestock grazing and soil-erosion control in arid and semiarid areas worldwide.Genomic information and molecular tools to support breeding and research in the species are limited.The objectives of this investigation were to map its genome using DNA markers and to identify quantitative trait loci(QTL)in the species.An F1 mapping population of 250 plants was developed from a cross between two parents with differing flowering-related traits.A high-density genetic linkage map containing 4821 markers on eight linkage groups(LGs)with a total genetic length of 1395 cM and a mean interval of 0.29 cM between adjacent markers was constructed with SLAF-seq technology.Comparative genomic analyses revealed the highest genome sequence similarity(8.71%)between erect milkvetch and Medicago truncatula,followed by Glycine max(7.65%),Cicer arietinum(7.53%),and Lupinus angustifolius(5.21%).A total of 64 significant QTL for flowering-related traits on six LGs were detected,accounting for 9.38 to 19.1%of the associated phenotype variation.Five and 48 key candidate genes for floret number and inflorescence length were identified based on the Glycyrrhiza uralensis genome.These candidate genes were involved in ubiquitination/degradation,pollen development,cell division,cytokinin biosynthetic process,and plant flowering.These findings shed light on the regulation of flowering traits in erect milkvetch and provide genomic resources for future molecular breeding of the crop.

Multiple phenotypes in genome-wide genetic mapping studies

For many psychiatric and other traits,diagnoses are based on a number of different criteria or phenotypes.Rather than carrying out genetic analyses on the final diagnosis,it has been suggested that relevant phenotypes should be analyzed directly.We provide an overview of statistical methods for the joint analysis of multiple phenotypes in case-control association studies.

Genetic analysis and QTL mapping of growth period traits and plant height traits in soybean recombinant inbred lines from Dongnong 47 × PI 317334-B

High and stable yield is the main goal of soybean genetic improvement.In this study,association analysis was used to detect the quantitative trait loci(QTL)for the plant height,and soybean growth period using 182 SSR markers in the RIL population of 136 F_(4:8) lines,which developed from a cross between photoperiod-insensitive cultivar‘Dongnong 47’and photoperiod-sensitive variety PI317334–B.The results showed that 33 QTLs related to soybean growth period and plant height traits were detected by compound interval mapping,and were located on 12 linkage groups including N,C1,C2,J,D1a,B2,E,G,A2,O,L,I,with the contribution rate of 7.85–33.84%.These QTL loci and linkage markers related to soybean photoperiod sensitivity,would be helpful to identify key genes that control soybean photoperiod sensitivity,and provide an important basis for the breeding of new photoperiod-insensitive soybean varieties based on molecular design breeding.

Identification and fine mapping of PmNJ3946 for powdery mildew resistance in einkorn wheat

Powdery mildew caused by Blumeria graminis f.sp.tritici(Bgt)is a destructive wheat disease.Although it can be easily overcome by deployment of resistance genes,the resistance is often quickly compromised by pathogen virulence.Thus,exploration and characterization of new resistance genes is always ongoing.Line NJ3946 derived from a cross of einkorn wheat accessions TA2032 and M389 showed resistance to powdery mildew.Inheritance analysis of an F2 population derived from a cross of NJ3946 and M389 suggested that the resistance was conferred by a dominant allele.With polymorphic markers identified through bulked segregant analysis(BSA),this gene was mapped to a novel locus on chromosome 3A,and was designated as PmNJ3946.Bulked segregant RNA-seq analysis(BSR-seq)was conducted to obtain more closely linked markers,which allowed delimitation of the PMNJ3946 locus to a 0.9 cM interval covering a physical distance of less than 1 Mb.PMNJ3946 was flanked by Xwgrc5153 and SNP-derived marker CHS21_3A008915069,and co-segregated with SNP-derived markers CHS21_3A008939814 and CHS21_3A008943175.The PmNJ3946 discovery expands the diversity of powdery mildew resistance genes and is useful for wheat breeding.

Genetic Map Pinpoints Elephant Poachers

美国华盛顿大学一个研究小组最近发表了最新的基因技术成果:通过遗传图谱描绘了非洲大陆各个地区象群遗传概貌的变化情况。这份图谱,不仅可以确定被查抄的非法象牙的出处,而且还将帮助各国的野生动植物保护官员对偷猎事件频发的地域进行重点管理,有效打击偷猎者。

Constructing the wolfberry(Lycium spp.) genetic linkage map using AFLP and SSR markers

Genetic linkage maps are important for quantitative trait locus(QTL) and marker-assisted selection breeding. The wolfberry(Lycium spp.) is an important food and traditional medicine in China. However, few construction genetic linkage maps have been reported because of the lack of genomic and genetic resources. In this study, a population of 89 F1 seedings was derived from a cross between two heterozygous parents, L. chinense var. potaninii ‘BF-01’(female)and L. barbarum var. auranticarpum ‘NH-01’(male), in order to construct a genetic linkage map using simple sequence repeat(SSR) and amplified fragment length polymorphism(AFLP) markers based on the double pseudo-test cross mapping strategy. The resulting genetic map consisted of 165 markers(74 AFLPs and 91 SSRs) distributed across 12 linkage groups and spanned a total length of 557.6 cM with an average distance of 3.38 cM between adjacent markers.The 12 linkage groups contained 3 to 21 markers and ranged in length from 8.6 to 58.3 cM. Twenty-nine segregated markers distributed in the map were mainly located on LG4 and LG9 linkage groups at P<0.05. This is the first linkage map of Lycium species using SSR and AFLP markers, which can serve as basis for improving genes and selective breeding of the genome assembly.

QTL Mapping for Fiber Quality Traits Based on a Dense Genetic Linkage Map with SSR,TRAP,SRAP and AFLP Markers in Cultivated Tetraploid Cotton

Cotton is one of the most important economic crops in the world,and it provides natural fiber for the textile industry.With the advancement of the textile technology and increased consumption demands on cotton fiber,both cotton yield and quality should be enhanced.However,cotton yield