Polymorphic Single Nucleotide Polymorphism Markers Research Articles

Identification of consistent QTL with large effect on anther extrusion in doubled haploid populations developed from spring wheat accessions in German Federal ex situ Genebank.

Novel large-effect consistent QTL for anther extrusion (AE) to improve cross-pollination were mapped in doubled haploid populations derived from IPK gene bank spring wheat accessions. TaAP2-D, an ortholog of Cleistogamy1 in barley, is a likely candidate gene for AE in wheat. To establish a robust hybrid wheat breeding system, male lines harboring alleles that promote outcrossing should be developed. In this study, we developed two doubled haploid (DH) populations of hexaploid spring wheat (Triticum aestivum L.) by crossing accessions taken from IPK gene bank. In both populations, the phenotypic data of anther extrusion (AE) based on three years of field trials showed a wide variation and approximated a normal distribution. Both populations were genotyped with a 15k Infinium single nucleotide polymorphism (SNP) array resulting in 3567 and 3457 polymorphic SNP markers for DH population-1 and DH population-2, respectively. Composite interval mapping identified quantitative trait loci (QTL) on chromosomes 1D, 2D, 4A, 4B, 5A,5D, 6A, and 6B; with consistent QTL (that are identified in all the years) on chromosome 4A in DH population-1, and on chromosomes 2D and 6B in DH population-2. The consistent QTL explained 17.2%, 32.9%, and 12.3% of thephenotypic variances, respectively. Genic scan of the chromosome 2D-QTL showed that the wheat gene TaAP2-D, an ortholog of Cleistogamy1 which promotes AE via swelling of the lodicules in barley, lies within the QTL region. A diagnostic marker was developed for TaAP2-D that showed co-segregation with the AE phenotype. This study shows the use of gene bank diversity reservoir to find alleles which are otherwise difficult to detect in elite populations. The identification of large-effect consistent QTL for AE is expected to help form efficient male parental lines suitable for hybrid wheat seed production and serve asa source for map-based cloning.

SNP marker development and polymorphism detection of Schizothorax waltoni

As a vulnerable species in the Red List of China’s Vertebrates, Schizothorax waltoni (S. waltoni) is mainly distributed in the middle reaches of Yarlung Tsangpo River in Tibet. Due to its delicate meat and delicious taste, S. waltoni is the main economic fish in the producing area. But recently, it has undergone a drastic population decline due to the overfishing of fishery resources, the invasion of foreign species, environmental pollution and other factors. Therefore, it is extremely urgent to protect germplasm resource of S. waltoni. In this study, we used Illumina Hiseq 4000 sequencing to develop single nucleotide polymorphism (SNP) markers in S. waltoni. Based on a large number of transcriptome sequences and candidate SNP loci obtained by previous transcriptome studies, 63 SNPs were randomly selected from immune-related genes and 36 loci exhibited significant polymorphisms in genotyping by Sequenom MassARRAY. The observed heterozygosity and expected heterozygosity varied from 0.0789 to 0.9577 and 0.0763 to 0.5033, respectively. There is only 5 SNP loci accord with Hardy–Weinberg equilibrium (p > 0.05) and no linkage disequilibria. These polymorphic SNP markers could be used for conservation genetics research in S. waltoni, and may provide new insights into future conservation strategies.

Utilization of the Genomewide Wheat 55K SNP Array for Genetic Analysis of Stripe Rust Resistance in Common Wheat Line P9936

Breeding for resistance to stripe rust (caused by Puccinia striiformis f. tritici) is essential for reducing losses in yield and quality in wheat. To identify genes for use in breeding, a biparental population of 186 recombinant inbred lines (RILs) from a cross of the Chinese landrace Mingxian 169 and CIMMYT-derived line P9936 was evaluated in field nurseries either artificially or naturally inoculated in two crop seasons. Each of the RILs and parents was genotyped with the wheat 55K single-nucleotide polymorphism (SNP) 'Breeders' array and a genetic linkage map with 8,225 polymorphic SNP markers spanning 3,593.37 centimorgans was constructed. Two major quantitative trait loci (QTL) and two minor QTL were identified. The major QTL QYr.nwafu-3BS.2 and QYr.nwafu-7BL on chromosomes arms 3BS and 7BL were detected in all field locations and explained an average 20.4 and 38.9% of phenotypic variation stripe rust severity, respectively. QYr.nwafu-3BS.2 likely corresponds to the locus Yr30/Sr2 and QYr.nwafu-7BL may be a resistance allele identified previously in CIMMYT germplasm. The other minor QTL had limited individual effects but increased resistance when in combinations with other QTL. Markers linked to QYr.nwafu-7BL were converted to kompetitive allele-specific polymerase chain reaction markers and validated in a panel of wheat accessions. Wheat accessions carrying the same haplotype as P9936 at the identified SNP loci had lower average stripe rust severity than the average severity of all other haplotypes.

High-density SNP linkage map construction and QTL mapping for flavonoid-related traits in a tea plant (Camellia sinensis) using 2b-RAD sequencing

BackgroundFlavonoids are important components that confer upon tea plants a unique flavour and health functions. However, the traditional breeding method for selecting a cultivar with a high or unique flavonoid content is time consuming and labour intensive. High-density genetic map construction associated with quantitative trait locus (QTL) mapping provides an effective way to facilitate trait improvement in plant breeding. In this study, an F1 population (LJ43×BHZ) was genotyped using 2b-restriction site-associated DNA (2b-RAD) sequencing to obtain massive single nucleotide polymorphism (SNP) markers to construct a high-density genetic map for a tea plant. Furthermore, QTLs related to flavonoids were identified using our new genetic map.ResultsA total of 13,446 polymorphic SNP markers were developed using 2b-RAD sequencing, and 4,463 of these markers were available for constructing the genetic linkage map. A 1,678.52-cM high-density map at an average interval of 0.40 cM with 4,217 markers, including 427 frameset simple sequence repeats (SSRs) and 3,800 novel SNPs, mapped into 15 linkage groups was successfully constructed. After QTL analysis, a total of 27 QTLs related to flavonoids or caffeine content (CAF) were mapped to 8 different linkage groups, LG01, LG03, LG06, LG08, LG10, LG11, LG12, and LG13, with an LOD from 3.14 to 39.54, constituting 7.5% to 42.8% of the phenotypic variation.ConclusionsTo our knowledge, the highest density genetic map ever reported was constructed since the largest mapping population of tea plants was adopted in present study. Moreover, novel QTLs related to flavonoids and CAF were identified based on the new high-density genetic map. In addition, two markers were located in candidate genes that may be involved in flavonoid metabolism. The present study provides valuable information for gene discovery, marker-assisted selection breeding and map-based cloning for functional genes that are related to flavonoid content in tea plants.

Linkage fine-mapping and QTLs affecting morpho-agronomic traits of a Mesoamerican × Andean RIL common bean population

This paper proposes the construction of a genetic linkage map with 376 recombinant inbred lines (RILs) derived from a cross between Mesoamerican × Andean common bean (Phaseolus vulgaris L) parents based on single nucleotide polymorphism (SNP) markers; and to detect quantitative trait loci (QTLs) associated with seven morpho-agronomic traits: number of days to flowering (DF), number of days to maturity (DM) or crop cycle; plant architecture (ARC); seed yield (YLD); degree of seed flatness (SF); seed shape (SS); and 100-seed weight (SW). A total of 3060 polymorphic SNP markers were used and 2041 segregated at a 1:1 ratio in the RIL population, as expected. These markers were subjected to linkage analysis in each chromosome. The genetic linkage analysis resulted in linkage maps with a total of 1962 markers spanning 1079.21 cM. A total of 29 QTLs associated with seven morpho-agronomic traits were detected on the 11 chromosomes, which explained between 3.83 and 32.92% of the phenotypic variation in DF. A total of 18 candidate genes associated with the detected QTLs were identified and related with biological processes, molecular functions and cellular components.

Genotyping of Soybean Cultivars With Medium-Density Array Reveals the Population Structure and QTNs Underlying Maturity and Seed Traits.

Soybean was domesticated about 5,000 to 6,000 years ago in China. Although genotyping technologies such as genotyping by sequencing (GBS) and high-density array are available, it is convenient and economical to genotype cultivars or populations using medium-density SNP array in genetic study as well as in molecular breeding. In this study, 235 cultivars, collected from China, Japan, USA, Canada and some other countries, were genotyped using SoySNP8k iSelect BeadChip with 7,189 single nucleotide polymorphisms (SNPs). In total, 4,471 polymorphic SNP markers were used to analyze population structure and perform genome-wide association study (GWAS). The most likely K value was 7, indicating this population can be divided into 7 subpopulations, which is well in accordance with the geographic origins of cultivars or accession studied. The LD decay rate was estimated at 184 kb, where r2 dropped to half of its maximum value (0.205). GWAS using FarmCPU detected a stable quantitative trait nucleotide (QTN) for hilum color and seed color, which is consistent with the known loci or genes. Although no universal QTNs for flowering time and maturity were identified across all environments, a total of 30 consistent QTNs were detected for flowering time (R1) or maturity (R7 and R8) on 16 chromosomes, most of them were corresponding to known E1 to E4 genes or QTL region reported in SoyBase (soybase.org). Of 16 consistent QTNs for protein and oil contents, 11 QTNs were detected having antagonistic effects on protein and oil content, while 4 QTNs soly for oil content, and one QTN soly for protein content. The information gained in this study demonstrated that the usefulness of the medium-density SNP array in genotyping for genetic study and molecular breeding.

GWAS for plant growth stages and yield components in spring wheat (Triticum aestivum L.) harvested in three regions of Kazakhstan

BackgroundSpring wheat is the largest agricultural crop grown in Kazakhstan with an annual sowing area of 12 million hectares in 2016. Annually, the country harvests around 15 million tons of high quality grain. Despite environmental stress factors it is predicted that the use of new technologies may lead to increases in productivity from current levels of 1.5 to up to 3 tons per hectare. One way of improving wheat productivity is by the application of new genomic oriented approaches in plant breeding projects. Genome wide association studies (GWAS) are emerging as powerful tools for the understanding of the inheritance of complex traits via utilization of high throughput genotyping technologies and phenotypic assessments of plant collections. In this study, phenotyping and genotyping data on 194 spring wheat accessions from Kazakhstan, Russia, Europe, and CIMMYT were assessed for the identification of marker-trait associations (MTA) of agronomic traits by using GWAS.ResultsField trials in Northern, Central and Southern regions of Kazakhstan using 194 spring wheat accessions revealed strong correlations of yield with booting date, plant height, biomass, number of spikes per plant, and number of kernels per spike. The accessions from Europe and CIMMYT showed high breeding potential for Southern and Central regions of the country in comparison with the performance of the local varieties. The GGE biplot method, using average yield per plant, suggested a clear separation of accessions into their three breeding origins in relationship to the three environments in which they were evaluated. The genetic variation in the three groups of accessions was further studied using 3245 polymorphic SNP (single nucleotide polymorphism) markers. The application of Principal Coordinate analysis clearly grouped the 194 accessions into three clades according to their breeding origins. GWAS on data from nine field trials allowed the identification of 114 MTAs for 12 different agronomic traits.ConclusionsField evaluation of foreign germplasm revealed its poor yield performance in Northern Kazakhstan, which is the main wheat growing region in the country. However, it was found that EU and CIMMYT germplasm has high breeding potential to improve yield performance in Central and Southern regions. The use of Principal Coordinate analysis clearly separated the panel into three distinct groups according to their breeding origin. GWAS based on use of the TASSEL 5.0 package allowed the identification of 114 MTAs for twelve agronomic traits. The study identifies a network of key genes for improvement of yield productivity in wheat growing regions of Kazakhstan.

Genome-wide association mapping reveals a rich genetic architecture of stripe rust resistance loci in emmer wheat (Triticum turgidum ssp. dicoccum)

Key messageSNP-based genome scanning in worldwide domesticated emmer germplasm showed high genetic diversity, rapid linkage disequilibrium decay and 51 loci for stripe rust resistance, a large proportion of which were novel.Cultivated emmer wheat (Triticum turgidum ssp. dicoccum), one of the oldest domesticated crops in the world, is a potentially rich reservoir of variation for improvement of resistance/tolerance to biotic and abiotic stresses in wheat. Resistance to stripe rust (Puccinia striiformis f. sp. tritici) in emmer wheat has been under-investigated. Here, we employed genome-wide association (GWAS) mapping with a mixed linear model to dissect effective stripe rust resistance loci in a worldwide collection of 176 cultivated emmer wheat accessions. Adult plants were tested in six environments and seedlings were evaluated with five races from the United States and one from Italy under greenhouse conditions. Five accessions were resistant across all experiments. The panel was genotyped with the wheat 90,000 Illumina iSelect single nucleotide polymorphism (SNP) array and 5106 polymorphic SNP markers with mapped positions were obtained. A high level of genetic diversity and fast linkage disequilibrium decay were observed. In total, we identified 14 loci associated with field resistance in multiple environments. Thirty-seven loci were significantly associated with all-stage (seedling) resistance and six of them were effective against multiple races. Of the 51 total loci, 29 were mapped distantly from previously reported stripe rust resistance genes or quantitative trait loci and represent newly discovered resistance loci. Our results suggest that GWAS is an effective method for characterizing genes in cultivated emmer wheat and confirm that emmer wheat is a rich source of stripe rust resistance loci that can be used for wheat improvement.

Genomewide Selection with Biallelic versus Triallelic Models in Three‐Way Maize Populations

While single nucleotide polymorphism (SNP) markers are typically biallelic, quantitative trait loci (QTL) may have three alleles per locus in three‐way populations. Our objective in this study was to determine if multiallelic markers or haplotypes improve the prediction accuracy of genomewide selection in three‐way breeding populations. Simulated and empirical maize (Zea mays L.) doubled haploid populations were used to compare a biallelic model, marker interval model (which used adjacent markers to create haplotypes), and allele phasing model (which inferred triallelic markers from parental SNP data). The simulation experiments differed in the number of QTL (10, 40, or 100), heritability (0.30, 0.50, or 0.80), and sizes of allelic effects. Four empirical three‐way populations were phenotyped at four to seven locations between 2012 and 2015 and were genotyped with 356 to 960 polymorphic SNP markers. Genomewide marker effects were obtained by ridge regression‐best linear unbiased prediction. In the simulation experiments, differences in prediction accuracy were <0.01 among the biallelic, marker interval, and allele phasing models. For grain yield, moisture, and test weight in the four maize populations, the differences in predictive ability among the three models were nonsignificant (P = 0.05). Further simulations showed that the small or nonsignificant differences in prediction accuracy were caused by large linkage blocks found among inbreds, particularly doubled haploids. Overall, we recommend the marker interval model in three‐way populations because of its simplicity, similar prediction accuracy, and theoretical advantage over the two other models.

Identification of QTL and Qualitative Trait Loci for Agronomic Traits Using SNP Markers in the Adzuki Bean.

The adzuki bean (Vigna angularis) is an important grain legume. Fine mapping of quantitative trait loci (QTL) and qualitative trait genes plays an important role in gene cloning, molecular-marker-assisted selection (MAS), and trait improvement. However, the genetic control of agronomic traits in the adzuki bean remains poorly understood. Single-nucleotide polymorphisms (SNPs) are invaluable in the construction of high-density genetic maps. We mapped 26 agronomic QTLs and five qualitative trait genes related to pigmentation using 1,571 polymorphic SNP markers from the adzuki bean genome via restriction-site-associated DNA sequencing of 150 members of an F2 population derived from a cross between cultivated and wild adzuki beans. We mapped 11 QTLs for flowering time and pod maturity on chromosomes 4, 7, and 10. Six 100-seed weight (SD100WT) QTLs were detected. Two major flowering time QTLs were located on chromosome 4, firstly VaFld4.1 (PEVs 71.3%), co-segregating with SNP marker s690-144110, and VaFld4.2 (PEVs 67.6%) at a 0.974 cM genetic distance from the SNP marker s165-116310. Three QTLs for seed number per pod (Snp3.1, Snp3.2, and Snp4.1) were mapped on chromosomes 3 and 4. One QTL VaSdt4.1 of seed thickness (SDT) and three QTLs for branch number on the main stem were detected on chromosome 4. QTLs for maximum leaf width (LFMW) and stem internode length were mapped to chromosomes 2 and 9, respectively. Trait genes controlling the color of the seed coat, pod, stem and flower were mapped to chromosomes 3 and 1. Three candidate genes, VaAGL, VaPhyE, and VaAP2, were identified for flowering time and pod maturity. VaAGL encodes an agamous-like MADS-box protein of 379 amino acids. VaPhyE encodes a phytochrome E protein of 1,121 amino acids. Four phytochrome genes (VaPhyA1, VaPhyA2, VaPhyB, and VaPhyE) were identified in the adzuki bean genome. We found candidate genes VaAP2/ERF.81 and VaAP2/ERF.82 of SD100WT, VaAP2-s4 of SDT, and VaAP2/ERF.86 of LFMW. A candidate gene VaUGT related to black seed coat color was identified. These mapped QTL and qualitative trait genes provide information helpful for future adzuki bean candidate gene cloning and MAS breeding to improve cultivars with desirable growth periods, yields, and seed coat color types.

Development of 34 SNP markers developed from the genome resequencing dataset in golden snub-nosed monkey (Rhinopithecus roxellana) using high resolution melting (HRM)

As single nucleotide polymorphisms (SNPs), known as the third-generation molecular markers, are extensively used in genetic studies. The rapid development of high-throughput sequencing technologies has enabled the development of a large number of SNP markers to be cost-effective and rapid. Based on the genome resequencing dataset of Rhinopithecus roxellana, we characterized 34 polymorphic SNP markers using a high resolution melting diversity assay. The observed heterozygosity varied from 0.323 to 0.725, while the expected heterozygosity ranged from 0.352 to 0.638. In addition, the MAF varied from 0.206 to 0.5000, while HWE values varied from 0.060 to 0.725. To the best of our knowledge, this is the largest set of SNP markers developed for R. roxellana by far, and could be useful in further geographic populations, analysis of evolutionary relationships and conservation genetics of this species.

Genome-Wide SNP Linkage Mapping and QTL Analysis for Fiber Quality and Yield Traits in the Upland Cotton Recombinant Inbred Lines Population.

It is of significance to discover genes related to fiber quality and yield traits and tightly linked markers for marker-assisted selection (MAS) in cotton breeding. In this study, 188 F8 recombinant inbred lines (RILs), derived from a intraspecific cross between HS46 and MARCABUCAG8US-1-88 were genotyped by the cotton 63K single nucleotide polymorphism (SNP) assay. Field trials were conducted in Sanya, Hainan Province, during the 2014–2015 cropping seasons under standard conditions. Results revealed significant differences (P < 0.05) among RILs, environments and replications for fiber quality and yield traits. Broad-sense heritabilities of all traits including fiber length, fiber uniformity, micronaire, fiber elongation, fiber strength, boll weight, and lint percentage ranged from 0.26 to 0.66. A 1784.28 cM (centimorgans) linkage map, harboring 2618 polymorphic SNP markers, was constructed, which had 0.68 cM per marker density. Seventy-one quantitative trait locus (QTLs) for fiber quality and yield traits were detected on 21 chromosomes, explaining 4.70∼32.28% phenotypic variance, in which 16 were identified as stable QTLs across two environments. Meanwhile, 12 certain regions were investigated to be involved in the control of one (hotspot) or more (cluster) traits, mainly focused on Chr05, Chr09, Chr10, Chr14, Chr19, and Chr20. Nineteen pairs of epistatic QTLs (e-QTLs) were identified, of which two pairs involved in two additive QTLs. These additive QTLs, e-QTLs, and QTL clusters were tightly linked to SNP markers, which may serve as target regions for map-based cloning, gene discovery, and MAS in cotton breeding.

SNP-based discovery of salinity-tolerant QTLs in a bi-parental population of rice (Oryza sativa).

Breeding for salt tolerance is the most promising approach to enhance the productivity of saline prone areas. However, polygenic inheritance of salt tolerance in rice acts as a bottleneck in conventional breeding for salt tolerance. Hence, we set our goals to construct a single nucleotide polymorphism (SNP)-based molecular map employing high-throughput SNP marker technology and to investigate salinity tolerant QTLs with closest flanking markers using an elite rice background. Seedling stage salinity responses were assessed in a population of 281 recombinant inbred lines (RILs) derived from the cross between At354 (salt tolerant) and Bg352 (salt susceptible), by 11 morpho-physiological indices under a hydroponic system. Selected extreme 94 RILs were genotyped using Illumina Infinium rice 6K SNP array and densely saturated molecular map spanning 1460.81cM of the rice genome with an average interval of 1.29cM between marker loci was constructed using 1135 polymorphic SNP markers. The results revealed 83 significant QTLs for 11 salt responsive traits explaining 12.5-46.7% of phenotypic variation in respective traits. Of them, 72 QTLs responsible for 10 traits were co-localized together forming 14 QTL hotspots at 14 different genomic regions. The all QTL hotspots were flanked less than 1Mb intervals and therefore the SNP loci associated with these QTL hotspots would be important in candidate gene discovery for salt tolerance.

Identification of SNP and SSR Markers in Finger Millet Using Next Generation Sequencing Technologies.

Finger millet is an important cereal crop in eastern Africa and southern India with excellent grain storage quality and unique ability to thrive in extreme environmental conditions. Since negligible attention has been paid to improving this crop to date, the current study used Next Generation Sequencing (NGS) technologies to develop both Simple Sequence Repeat (SSR) and Single Nucleotide Polymorphism (SNP) markers. Genomic DNA from cultivated finger millet genotypes KNE755 and KNE796 was sequenced using both Roche 454 and Illumina technologies. Non-organelle sequencing reads were assembled into 207 Mbp representing approximately 13% of the finger millet genome. We identified 10,327 SSRs and 23,285 non-homeologous SNPs and tested 101 of each for polymorphism across a diverse set of wild and cultivated finger millet germplasm. For the 49 polymorphic SSRs, the mean polymorphism information content (PIC) was 0.42, ranging from 0.16 to 0.77. We also validated 92 SNP markers, 80 of which were polymorphic with a mean PIC of 0.29 across 30 wild and 59 cultivated accessions. Seventy-six of the 80 SNPs were polymorphic across 30 wild germplasm with a mean PIC of 0.30 while only 22 of the SNP markers showed polymorphism among the 59 cultivated accessions with an average PIC value of 0.15. Genetic diversity analysis using the polymorphic SNP markers revealed two major clusters; one of wild and another of cultivated accessions. Detailed STRUCTURE analysis confirmed this grouping pattern and further revealed 2 sub-populations within wild E. coracana subsp. africana. Both STRUCTURE and genetic diversity analysis assisted with the correct identification of the new germplasm collections. These polymorphic SSR and SNP markers are a significant addition to the existing 82 published SSRs, especially with regard to the previously reported low polymorphism levels in finger millet. Our results also reveal an unexploited finger millet genetic resource that can be included in the regional breeding programs in order to efficiently optimize productivity.

Genome-Wide QTL Mapping for Wheat Processing Quality Parameters in a Gaocheng 8901/Zhoumai 16 Recombinant Inbred Line Population.

Dough rheological and starch pasting properties play an important role in determining processing quality in bread wheat (Triticum aestivum L.). In the present study, a recombinant inbred line (RIL) population derived from a Gaocheng 8901/Zhoumai 16 cross grown in three environments was used to identify quantitative trait loci (QTLs) for dough rheological and starch pasting properties evaluated by Mixograph, Rapid Visco-Analyzer (RVA), and Mixolab parameters using the wheat 90 and 660 K single nucleotide polymorphism (SNP) chip assays. A high-density linkage map constructed with 46,961 polymorphic SNP markers from the wheat 90 and 660 K SNP assays spanned a total length of 4121 cM, with an average chromosome length of 196.2 cM and marker density of 0.09 cM/marker; 6596 new SNP markers were anchored to the bread wheat linkage map, with 1046 and 5550 markers from the 90 and 660 K SNP assays, respectively. Composite interval mapping identified 119 additive QTLs on 20 chromosomes except 4D; among them, 15 accounted for more than 10% of the phenotypic variation across two or three environments. Twelve QTLs for Mixograph parameters, 17 for RVA parameters and 55 for Mixolab parameters were new. Eleven QTL clusters were identified. The closely linked SNP markers can be used in marker-assisted wheat breeding in combination with the Kompetitive Allele Specific PCR (KASP) technique for improvement of processing quality in bread wheat.

Construction of a high-density genetic map of Ziziphus jujuba Mill. using genotyping by sequencing technology

The Chinese jujube (Ziziphus jujuba Mill., 2n = 2 × = 24), one of the most popular fruit trees in China, is widely cultivated and utilized in Asia. High-density genetic linkage maps are valuable resources for molecular breeding and functional genomics; however, they are still under-developed for the jujube. The genotyping by sequencing (GBS) strategy could be an efficient and cost-effective tool for single nucleotide polymorphism (SNP) discovery based on the sequenced jujube genome. Here, we report a new high-density genetic map constructed using GBS technology. An F1 population with 145 progenies and their parents (‘Dongzao’ × ‘Zhongningyuanzao’) were sequenced on the Illumina HiSeq 4000 platform. In total, 79.8 Gb of raw data containing 256,708,177 paired-end reads were generated. After data filtering and SNP genotyping, 40,372 polymorphic SNP markers were developed between the parents and 2540 (1756 non-redundant) markers were mapped onto the integrated genetic linkage map. The map spanned 1456.53 cM and was distributed among 12 linkage groups, which is consistent with the haploid chromosome number of the jujube. The average marker interval was 0.88 cM. The genetic map allowed us to anchor 224 Mb (63.7 %) of scaffolds from the sequenced ‘Junzao’ genome, containing 52 newly anchored scaffolds, which extended the genome assembly by 7 Mb. In conclusion, GBS technology was applied efficiently for SNP discovery in this study. The high-density genetic map will serve as a unique tool for molecular-assisted breeding and genomic studies, which will contribute to further research and improvement of the jujube in the near future.

English

Genetic linkage maps provide a genomic framework for quantitative trait loci identification applied in marker assisted selection breeding in crops with limited resources. It serves as a powerful tool to breeders for analysing the mode of inheritance of genes of interest and monitoring of the transmission of target genes from parents to progeny. Single nucleotide polymorphism (SNP) markers have a great influence on mapping research studies: for faster development of new varieties of crops due to its abundance and high number of polymorphism. Thus, this research work aimed at constructing genetic linkage map of cowpea using SNP markers. This will facilitate development of new varieties of cowpea. Single seed descent method of breeding was used to generate mapping population between IT-95K-193-12 and Ife-Brown. Fresh leaves from both parental lines and F1 hybrids were genotyped with 135 SNP markers. Data from125 polymorphic SNP markers were used to construct linkage map. Results revealed 12 linkage maps with an average of 10 markers to a chromosome and average marker distance of 3.02 cM between the markers. Constructed map provides basic information that could assist in genetic improvement of cowpea, most especially in developing brown blotch resistant cowpea varieties through detection of quantitative trait loci (QTL) responsible for brown blotch resistance in cowpea when utilizing marker assisted selection method. Key words: Cowpea, genetic linkage map, single nucleotide polymorphism (SNP) markers, brown blotch resistance.

Genome-wide linkage mapping of QTL for physiological traits in a Chinese wheat population using the 90K SNP array

Many physiological traits are associated with grain yield and yield-related traits in wheat. The aim of the present study was to identify quantitative trait loci (QTL) for physiological traits, including ground cover, normalized difference in vegetation index and canopy temperature depression, and closely linked molecular markers for marker-assisted breeding. Two hundred and forty-six F8 recombinant inbred lines (RILs) come from the Zhou 8425B/Chinese Spring cross were genotyped using the high-density Illumina iSelect 90K single nucleotide polymorphism (SNP) assay. Field trials were conducted under irrigated conditions at Zhengzhou and Zhoukou in Henan province, during the 2012–2013 and 2013–2014 cropping seasons. Analysis of variance of physiological traits showed significant differences (P < 0.01) among RILs, and RILs × environment interactions. Ground cover at late spring (GC-S), normalized difference in vegetation index at late spring (NDVI-S), and ground cover pre-winter (GC-W), had moderate broad-sense heritabilities of 0.66, 0.52 and 0.40, respectively. GC-S, NDVI-S and canopy temperature depression at 10 days post anthesis (CTD-10) were positively correlated with GY. Using a high-density linkage map of 5636 polymorphic SNP markers and composite interval mapping, 24 QTL for GC, NDVI and CTD were identified on 12 chromosomes, explaining 3.4–14.6 % of the phenotypic variance. Five stable QTL were detected across three environments, viz. QGC-W.caas-7AL, QNDVI-S.caas-7AL, QGC-S.caas-3AS, QCTD-A.caas-5BS and QCTD-10.caas-5BS. In addition, 10 QTL clusters were observed on chromosomes 1AL, 2AL, 2BL, 3AS, 3B (2), 4BS, 5B, 7AS, and 7AL. The stable QTL and QTL clusters were linked to SNP markers, with genetic distances to the closest SNPs ranging from 0 to 2.0 cM; these could be used for marker-assisted selection to improve yield-related traits in wheat breeding.

Analysis of genetic diversity and population structure in a tomato (Solanum lycopersicum L.) germplasm collection based on single nucleotide polymorphism markers.

Knowledge of genetic diversity is important to assist breeders in the selection of parental materials and in the design of breeding programs. In this study, we genotyped 348 inbred tomato lines, representing vintage and contemporary fresh-market varieties, by using 52 single nucleotide polymorphisms (SNPs); 45 of these were found to be polymorphic. The average minor allele frequency and unbiased expected heterozygosity were 0.315 and 0.356, respectively. Population structure analysis revealed that contemporary germplasm could be distinctly divided into six subpopulations representing three market classes and breeding programs (pink, green, and red). Vintage germplasm could be separated into at least two subpopulations, and more admixtures were found in vintage lines than in contemporary lines. These findings indicate that contemporary inbred lines are more diversified than vintage inbred lines. AMOVA of vintage and contemporary lines was performed. A significant difference was found (P < 0.01), which explained 17.4% of the total genetic variance. Subsequently, we constructed a core collection using 45 polymorphic SNP markers. The data showed that all alleles were captured by only 2% of lines, indicating that more alleles, as well as rare alleles, could enable more variation to be captured in the core collection. These data allow us to discard redundant inbred tomato lines and to select elite inbred lines, which will accelerate the breeding process.

Genome-Wide Linkage Mapping of QTL for Yield Components, Plant Height and Yield-Related Physiological Traits in the Chinese Wheat Cross Zhou 8425B/Chinese Spring.

Identification of genes for yield components, plant height (PH), and yield-related physiological traits and tightly linked molecular markers is of great importance in marker-assisted selection (MAS) in wheat breeding. In the present study, 246 F8 RILs derived from the cross of Zhou 8425B/Chinese Spring were genotyped using the high-density Illumina iSelect 90K single nucleotide polymorphism (SNP) assay. Field trials were conducted at Zhengzhou and Zhoukou of Henan Province, during the 2012–2013 and 2013–2014 cropping season under irrigated conditions, providing data for four environments. Analysis of variance (ANOVA) of agronomic and physiological traits revealed significant differences (P < 0.01) among RILs, environments, and RILs × environments interactions. Broad-sense heritabilities of all traits including thousand kernel weight (TKW), PH, spike length (SL), kernel number per spike (KNS), spike number/m2 (SN), normalized difference in vegetation index at anthesis (NDVI-A) and at 10 days post-anthesis (NDVI-10), SPAD value of chlorophyll content at anthesis (Chl-A) and at 10 days post-anthesis (Chl-10) ranged between 0.65 and 0.94. A linkage map spanning 3609.4 cM was constructed using 5636 polymorphic SNP markers, with an average chromosome length of 171.9 cM and marker density of 0.64 cM/marker. A total of 866 SNP markers were newly mapped to the hexaploid wheat linkage map. Eighty-six QTL for yield components, PH, and yield-related physiological traits were detected on 18 chromosomes except 1D, 5D, and 6D, explaining 2.3–33.2% of the phenotypic variance. Ten stable QTL were identified across four environments, viz. QTKW.caas-6A.1, QTKW.caas-7AL, QKNS.caas-4AL, QSN.caas-1AL.1, QPH.caas-4BS.2, QPH.caas-4DS.1, QSL.caas-4AS, QSL.caas-4AL.1, QChl-A.caas-5AL, and QChl-10.caas-5BL. Meanwhile, 10 QTL-rich regions were found on chromosome 1BS, 2AL (2), 3AL, 4AL (2), 4BS, 4DS, 5BL, and 7AL exhibiting pleiotropic effects. These QTL or QTL clusters are tightly linked to SNP markers, with genetic distances to the closest SNPs ranging from 0 to 1.5 cM, and could serve as target regions for fine mapping, candidate gene discovery, and MAS in wheat breeding.