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Genotyping‐by‐sequencing (GBS) is a widely used strategy for obtaining large numbers of genetic markers in model and non‐model organisms. In crop plants, GBS‐derived marker datasets are frequently used to perform quantitative trait locus (QTL) mapping. In some plant species, however, high heterozygosity and complex genome structure mean that researchers must use care in handling GBS data to conduct QTL mapping most effectively. Such outbred crops include most of the perennial grass and tree species used for bioenergy. To identify strategies for increasing accuracy and precision of QTL mapping using GBS data in outbred crops, we conducted an empirical study of SNP‐calling and genetic map‐building pipeline parameters in a Miscanthus sinensis population, and a complementary simulation study to estimate the relationship between genome‐wide error rate, read depth, and marker number. The bioenergy grass Miscanthus is an obligate outcrossing species with a recent (diploidized) whole‐genome duplication. For the study of empirical M. sinensis data, we compared two SNP‐calling methods (one non‐reference‐based and one reference‐based), a series of depth filters (12×, 20×, 30×, and 40×) and two map‐construction methods (i.e., marker ordering: linkage‐only and order‐corrected based on a reference genome). We found that correcting the order of markers on a linkage map by using a high‐quality reference genome improved QTL precision (shorter confidence intervals). For typical GBS datasets of between 1000 and 5000 markers to build a genetic map for biparental populations, a depth filter set at 30× to 40× applied to outbred populations provided a genome‐wide genotype‐calling error rate of less than 1%, improved accuracy of QTL point estimates and minimized type I errors for identifying QTL. Based on these results, we recommend using a reference genome to correct the marker order of genetic maps and a robust genotype depth filter to improve QTL mapping for outbred crops. [ABSTRACT FROM AUTHOR]

Tools for visualizing genomes are essential for investigating genomic features and their interactions. Currently, tools designed originally for animal mitogenomes and plant plastomes are used to visualize the mitogens of plants but cannot accurately display features specific to plant mitogenomes, such as nonlinear exon arrangement for genes, the prevalence of functional noncoding features and complex chromosomal architecture. To address these problems, a software package, plant mitochondrial genome map (PMGmap), was developed using the Python programming language. PMGmap can draw genes at exon levels; draw cis‐ and trans‐splicing gene maps, noncoding features and repetitive sequences; and scale genic regions by using the scaling of the genic regions on the mitogenome (SAGM) algorithm. It can also draw multiple chromosomes simultaneously. Compared with other state‐of‐the‐art tools, PMGmap showed better performance in visualizing 405 plant mitogenomes, showing potential as an invaluable tool for plant mitogenome research. The web and container versions and the source code of PMGmap can be accessed through the following link: http://www.1kmpg.cn/pmgmap. [ABSTRACT FROM AUTHOR]

The highly unique zigzag-shaped stem phenotype in tea plants boasts significant ornamental value and is exceptionally rare. To investigate the genetic mechanism behind this trait, we developed BC1 artificial hybrid populations. Our genetic analysis revealed the zigzag-shaped trait as a qualitative trait. Utilizing whole-genome resequencing, we constructed a high-density genetic map from the BC1 population, incorporating 5,250 SNP markers across 15 linkage groups, covering 3,328.51 cM with an average marker interval distance of 0.68 cM. A quantitative trait locus (QTL) for the zigzag-shaped trait was identified on chromosome 4, within a 61.2 to 97.2 Mb range, accounting for a phenotypic variation explained (PVE) value of 13.62%. Within this QTL, six candidate genes were pinpointed. To better understand their roles, we analyzed gene expression in various tissues and individuals with erect and zigzag-shaped stems. The results implicated CsXTH (CSS0035625) and CsCIPK14 (CSS0044366) as potential key contributors to the zigzag-shaped stem formation. These discoveries lay a robust foundation for future functional genetic mapping and tea plant genetic enhancement. [ABSTRACT FROM AUTHOR]

Background: Genome Mapping Technologies (optical and electronic) use ultra-high molecular weight DNA to detect structural variation and have application in constitutional genetic disorders, hematological neoplasms, and solid tumors. Genome mapping can detect balanced and unbalanced structural variation, copy number changes, and haplotypes. The technique is analogous to chromosomal microarray analysis, although genome mapping has the added benefit of being able to detect and ascertain the nature of more abnormalities in a single assay than array, karyotyping, or FISH alone. Key Messages: This paper describes a specific nomenclature for genome mapping that can be used by diagnostic and research centers to report their findings accurately. An international nomenclature is essential for patient results to be understood by different healthcare providers as well as for clear communication in publications and consistency in databases. Summary: Genome mapping can detect aneuploidy, balanced and unbalanced structural variation, as well as copy number changes. The Standing Committee for the International System for Human Cytogenomic Nomenclature (ISCN) recognised there was a need for a specific nomenclature for genome mapping that encompasses the range of abnormalities detected by this technique. This paper explains the general principles of the nomenclature as well as giving specific ISCN examples for the different types of numerical and structural rearrangements. [ABSTRACT FROM AUTHOR]

Background: Growth rate is a crucial economic trait for farmed animals, but the genetic regulation of this trait is largely unknown in non-model organisms such as shrimp. Results: In this study, we performed genome-wide phenotypic quantitative trait loci (QTL) and expression quantitative trait loci (eQTL) mapping analyses to identify genes affecting the growth rate of Pacific white shrimp (Litopenaeus vannamei), which is the most commercially-farmed crustacean worldwide. We used RNA-sequencing of 268 individuals in a mapping population, and subsequently validated our findings through gene silencing and shrimp growth experiments. We constructed a high-density genetic linkage map comprising 5533 markers spanning 44 linkage groups, with a total distance of 6205.75 cM and an average marker interval of 1.12 cM. Our analyses identified 11 QTLs significantly correlated with growth rate, and 117,525 eQTLs. By integrating QTL and eQTL data, we identified a gene (metalloreductase STEAP4) highly associated with shrimp growth rate. RNA interference (RNAi) analysis and growth experiments confirmed that STEAP4 was significantly correlated with growth rate in L. vannamei. Conclusions: Our results indicate that the comprehensive analysis of QTL and eQTL can effectively identify genes involved in complex animal traits. This is important for marker-assisted selection (MAS) of animals. Our work contributes to the development of shrimp breeding and available genetic resources. [ABSTRACT FROM AUTHOR]

Flower colour, as an important morphological marker, plays an essential role in improving the identification efficiency of the purity seed in hybrid production. However, the molecular mechanism of white‐flower trait has not been reported in pumpkin (Cucurbita moschata D.). In this study, we obtained a white‐flower mutant (wf) through the ethyl methane sulfonate (EMS) mutagenesis of inbred line N87 (yellow flower). F2 populations were then constructed by crossing wf mutant and N87 plant to fine map the genes controlling white‐flower trait in pumpkin. Phenotypic identification revealed that carotenoid content significantly decreased in the petals of wf mutants compared with N87 plants. Genetic analysis indicated that the white flower mutant trait was controlled by a single recessive gene, Cmowf. Using bulked segregant analysis and KASP phenotyping, Cmowf was mapped to a 762 kb region on chromosome 14 containing three annotated genes. Among them, a nonsynonymous single‐nucleotide polymorphisms mutation was identified only in CmoCh14G005820 gene, which encoded a DUF1997 family protein. Compared with CmoDUF1997 amino acid sequences between the wf mutants and N87 plants, the critical amino acid mutations (early termination of amino acids) occurred in wf mutants, so CmoCh14G005820 was predicted as a potential candidate for controlling the white‐flower trait. RNA‐sequencing analysis revealed that the expression of CmoCh14G005820 and most genes involved in carotenoid biosynthesis was significantly downregulated in wf mutants, whereas the expression of several genes responsible for carotenoid degradation was upregulated in wf mutants. This finding suggested that carotenoid metabolism may participate in the formation of flower colour in pumpkin. Overall, our results provided a theoretical basis for understanding the genetic mechanisms underlying white‐flower formation in pumpkin. [ABSTRACT FROM AUTHOR]

As an important spice species in Rutaceae, the Sichuan pepper (Zanthoxylum armatum) can provide pungent and numbing taste, as well as aroma in its mature fruit. Here we assembled a chromosome-level genome of green prickly ash which was widely cultivated in a major production area including Chongqing and Sichuan province, China. We generated 712 Gb (~112 x) PacBio long reads and 511 Gb (~82 x) Hi-C data, and yielded an assembly of 99 pseudochromosomes with total size of 5.32 Gb and contig N50 of 796 kb. The genomic analyses and cytogenetic experiments both indicated that the cultivar 'Zhuye Huajiao' was a triploid. We identified a Zanthoxylum-specific whole genome duplication event emerging about 24.8 million years ago (Mya). We also detected a transposition burst event (0.3e0.4 Mya) responsible for the large genome size of Z. armatum. Metabolomic analysis of the Zanthoxylum fruits during development stages revealed profiles of 39 volatile aroma compounds and 528 secondary metabolites, from which six types of sanshools were identified. Based on metabolomic and transcriptomic network analysis, we screened candidate genes encoding long chain acyl-CoA synthetase, fatty acid desaturase, branched-chain amino acid aminotransferase involved in sanshool biosynthesis and three genes encoding terpene synthase during fruit development. The multi-omics data provide insights into the evolution of Zanthoxylum and molecular basis of numbing and aroma flavor of Sichuan pepper. [ABSTRACT FROM AUTHOR]

Despite the longstanding importance of silage as a critical feed source for ruminants, its quality improvement has been largely overlooked. Although numerous quantitative trait loci (QTL) and genes affecting silage quality in maize have been reported, only a few have been effectively incorporated into breeding programs. Addressing this gap, the present study undertook a comprehensive meta-QTL (MQTL) analysis involving 523 QTL associated with silage-quality traits collected from 14 published studies. Of the 523 QTL, 405 were projected onto a consensus map comprising 62,424 genetic markers, resulting in the identification of 60 MQTL and eight singletons. The average confidence interval (CI) of the MQTL was 3.9-fold smaller than that of the source QTL. Nine of the 60 identified MQTL were classified as breeder’s MQTL owing to their small CIs, involvement of more QTL, and large contribution to phenotypic variation. One-third of the MQTL co-localized with DNA marker-trait associations identified in previous genomewide association mapping studies. A set of 78 high-confidence candidate genes influencing silage quality were identified in the MQTL regions. These genes and associated markers may advance marker-assisted breeding for maize silage quality. [ABSTRACT FROM AUTHOR]

More than 170 years after causing the potato famine in Ireland, late blight is still considered one of the most devastating crop diseases. Commercial potato breeding efforts depend on natural sources of resistance to protect crops from the rapidly evolving late blight pathogen, Phytophthora infestans. We have identified and mapped a novel broad-spectrum disease resistance gene effective against P. infestans from the wild, diploid potato species Solanum bulbocastanum. Diagnostic resistance gene enrichment sequencing (dRenSeq) was used to confirm the uniqueness of the identified resistance. RenSeq and GenSeq-based mapping of the resistance, referred to as Rpi-blb4, alongside recombinant screening, positioned the locus responsible for the resistance to potato chromosome 5. The interval spans approximately 2.3 Mb and corresponds to the DM reference genome positions of 11.25 and 13.56 Mb. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), threatens global wheat production. Development of cultivars with increased resistance to stem rust by identification, mapping, and deployment of resistance genes is the best strategy for controlling the disease. In this study, we performed fine mapping and characterization of the all-stage stem rust resistance (Sr) gene Sr8155B1 from the durum wheat line 8155-B1. In seedling tests of biparental populations, Sr8155B1 was effective against six Chinese Pgt races tested. In a segregating population of 5060 gametes, Sr8155B1 was mapped to a 0.06-cM region flanked by markers Pku2772 and Pku43365, corresponding to 1.5- and 2.7-Mb regions in the Svevo and Chinese Spring reference genomes. Both regions include several typical nucleotide-binding leucine-rich repeat (NLR) and protein kinase genes that represent candidate genes. Among them, three NLR genes and three receptor-like protein kinases were highly polymorphic between the parental lines and their transcripts were upregulated in the homozygous resistant line TdR2 relative to its susceptible sister line TdS4. Four markers (Pku2772, Pku43365, Pku2950, and Pku3721) developed in this study, together with seedling resistance responses, correctly predicted Sr8155B1 absence or presence in 78 tetraploid wheat genotypes tested. The presence of Sr8155B1 in tetraploid wheat accessions CItr 14916, PI 197492, and PI 197493 was confirmed by mapping in three F2 populations. The genetic map and linked markers developed in this study may accelerate the deployment of Sr8155B1-mediated resistance in wheat breeding programs. [ABSTRACT FROM AUTHOR]

High-quality molecular markers are essential for marker-assisted selection to accelerate breeding progress. Compared with diploid species, recently diverged polyploid crop species tend to have highly similar homeologous subgenomes, which is expected to limit the development of broadly applicable locus-specific single-nucleotide polymorphism (SNP) assays. Furthermore, it is particularly challenging to make genome-wide marker sets for species that lack a reference genome. Here, we report the development of a genome-wide set of kompetitive allele specific PCR (KASP) markers for marker-assisted recurrent selection (MARS) in the tetraploid minor crop perilla. To find locus-specific SNP markers across the perilla genome, we used genotyping-by-sequencing (GBS) to construct linkage maps of two F2 populations. The two resulting high-resolution linkage maps comprised 2326 and 2454 SNP markers that spanned a total genetic distance of 2133 cM across 16 linkage groups and 2169 cM across 21 linkage groups, respectively. We then obtained a final genetic map consisting of 22 linkage groups with 1123 common markers from the two genetic maps. We selected 96 genome-wide markers for MARS and confirmed the accuracy of markers in the two F2 populations using a high-throughput Fluidigm system. We confirmed that 91.8% of the SNP genotyping results from the Fluidigm assay were the same as the results obtained through GBS. These results provide a foundation for marker-assisted backcrossing and the development of new varieties of perilla. [ABSTRACT FROM AUTHOR]

Quantitative trait locus (QTL) mapping based on a genetic map is a very effective method of marker-assisted selection in breeding, and whole-genome resequencing is one of the useful methods to obtain high-density genetic maps. In this study, the hybrid assembly of Illumina, PacBio, and chromatin interaction mapping data was used to construct high-quality chromosomal genome sequences of Paulownia fortunei, with a size of 476.82 Mb, a heterozygosity of 0.52%, and a contig and scaffold N50s of 7.81 Mb and 21.81 Mb, respectively. Twenty scaffolds with a total length of 437.72 Mb were assembled into 20 pseudochromosomes. Repeat sequences with a total length of 243.96 Mb accounted for 51.16% of the entire genome. In all, 26,903 protein-coding gene loci were identified, and 26,008 (96.67%) genes had conserved functional motifs. Further comparative genomics analysis preliminarily showed that the split of P. fortunei with Tectona grandis likely occurred 38.8 (33.3–45.1) million years ago. Whole-genome resequencing was used to construct a merged genetic map of 20 linkage groups, with 2993 bin markers (3,312,780 SNPs), a total length of 1675.14 cm, and an average marker interval of 0.56 cm. In total, 73 QTLs for important phenotypic traits were identified (19 major QTLs with phenotypic variation explained ≥ 10%), including 10 for the diameter at breast height, 7 for the main trunk height, and 56 for branch-related traits. These results not only enrich P. fortunei genomic data but also form a solid foundation for fine QTL mapping and key marker/gene mining of Paulownia, which is of great significance for the directed genetic improvement of these species. [ABSTRACT FROM AUTHOR]

Ornamental kale (Brassica oleracea var. acephala) is an attractive ornamental plant with a range of leaf colors and shapes. Breeding new varieties of ornamental kale has proven challenging due to its lengthy breeding cycle and the limited availability of genetic markers. In this study, a F1DH ornamental kale population comprising 300 DH lines was constructed using microspore culture. A high-density genetic map was developed by conducting whole-genome sequencing on 150 individuals from the F1DH population. The genetic map contained 1696 bin markers with 982,642 single-nucleotide polymorphisms (SNPs) spanning a total distance of 775.81 cM on all nine chromosomes with an average distance between markers of 0.46 cM. The ornamental kale genetic map contained substantially more SNP markers compared with published genetic maps for other B. oleracea crops. Furthermore, utilizing this high-density genetic map, we identified seven quantitative trait loci (QTLs) that significantly influence the leaf shape of ornamental kale. These findings are valuable for understanding the genetic basis of key agronomic traits in ornamental kale. The F1DH progenies provide an excellent resource for germplasm innovation and breeding new varieties of ornamental kale. Additionally, the high-density genetic map provides crucial insights for gene mapping and unraveling the molecular mechanisms behind important agronomic traits in ornamental kale. [ABSTRACT FROM AUTHOR]

Takifugu fasciatus is an aquaculture species with high economic value. In recent years, problems such as environmental pollution and inbreeding have caused a serious decline in T. fasciatus germplasm resources. In this study, a high-density genetic linkage map was constructed by whole-genome resequencing. The map consists of 4891 bin markers distributed across 22 linkage groups (LGs), with a total genetic coverage of 2381.353 cM and a mean density of 0.535 cM. Quantitative trait locus (QTL) localization analysis showed that a total of 19 QTLs associated with growth traits of T. fasciatus in the genome-wide significance threshold range, distributed on 11 LGs. In addition, 11 QTLs associated with cold tolerance traits were identified, each scattered on a different LG. Furthermore, we used QTL localization analysis to screen out three candidate genes (IGF1, IGF2, ADGRB) related to growth in T. fasciatus. Meanwhile, we screened three candidate genes (HSP90, HSP70, and HMGB1) related to T. fasciatus cold tolerance. Our study can provide a theoretical basis for the selection and breeding of cold-tolerant or fast-growing T. fasciatus. [ABSTRACT FROM AUTHOR]

Single nucleotide polymorphism (SNP) genotyping arrays provide an optimal high-throughput platform for genetic research and molecular breeding programs in both animals and plants. In this study, a highquality and custom-designed Rice3K56 SNP array was developed with the resequencing data of 3024 rice accessions worldwide, which was then tested extensively in 192 representative rice samples. Printed on the GeneTitan chips of Affymetrix Axiom each containing 56,606 SNP markers, the Rice3K56 array has a high genotyping reliability (99.6%), high and uniform genome coverage (an average of 6.7-kb between adjacent SNPs), abundant polymorphic information and easy automation, compared with previously developed rice SNP arrays. When applied in rice varietal differentiation, population diversity analysis, gene mapping of 13 complex traits by a genome-wide association study analysis (GWAS), and genome selection experiments in a recombinant inbred line and a multi-parent advanced generation inter-cross populations, these properties of the Rice3K56 array were well demonstrated for its power and great potential to be a highly efficient tool for rice genetic research and genomic breeding. [ABSTRACT FROM AUTHOR]

The wild abortive (WA)-type cytoplasmic male sterility (CMS) derived from the wild rice species Oryza rufipogon Griff. is used widely in three-line indica hybrids. The identification and mapping of restorer of fertility (Rf) genes aided in the development of WA-type hybrids. Here we report that testcross F1 plants from the WA-type CMS line and 9311 exhibited stainable pollen grains with no seed set, indicating that 9311 carries minor-effect Rfs for WA-type CMS. We developed an advanced backcross population consisting of plants harboring small regions of donor chromosomal segments from 9311 in the WATianfengA genetic background with moderate seed setting rates. Genetic analysis showed that the pollen fertility levels of the backcross individuals are governed by a single gene from 9311 that we named Rf19 (t). By use of the RICE 40 K gene chip, three introduced segments were identified in the fertile lines, and a candidate region spanning 4.37–8.29 Mb on chromosome 1 was identified for Rf19(t). Finally, Rf19(t) was fine-mapped to a region of 90 kb between the DNA marker loci STS1-163 and STS1-183, in which eight ORFs were predicted. Also, using relative expression analyses, comparative sequence analyses and functional domain analyses, we identified LOC_Os01g10530 as the most likely candidate gene for Rf19(t). Furthermore, Rf19(t) was found to function in fertility restoration, most probably by regulating the degradation of mRNA transcribed from the mitochondrial gene WA352. These results increase our knowledge of fertility restoration in WA-type CMS lines and will facilitate the development of high-quality pairs of WAtype CMS and maintainer lines. [ABSTRACT FROM AUTHOR]

Apple replant disease (ARD) is a worldwide economic risk in apple production. Although several studies have shown that the wild apple accession Malus × robusta 5 (Mr5) is ARD-tolerant, the genetics of this tolerance have not yet been elucidated. A genetic mapping approach with a biparental population derived from contrasting parents involving molecular markers provides a means for marker-assisted selection of genetically complex traits and for determining candidate genes. In this study, we crossed the ARD-tolerant wild apple accession Mr5 and the ARD-susceptible rootstock 'M9' and analyzed the resultant progeny for ARD tolerance. Hence, a high-density genetic map using a tunable genotyping-by-sequencing (tGBS) approach was established. A total of 4804 SNPs together with 77 SSR markers were included in the parental maps comprising 17 linkage groups. The phenotypic responses to ARD were evaluated for 106 offspring and classified by an ARD-susceptibility index (ASI). A Kruskal–Wallis test identified SNP markers and one SSR marker on linkage groups (LG) 6 and 2 that correlated with ARD tolerance. We found nine candidate genes linked with these markers, which may be associated with plant response to ARD. These candidate genes provide some insight into the defense mechanisms against ARD and should be studied in more detail. [ABSTRACT FROM AUTHOR]

Leaf rust caused by Puccinia triticina Erikss. (Pt) is the most widely distributed and important wheat disease worldwide. The objective of the present study was to determine the frequency of Iranian Pt races, their virulence to key resistance genes and map quantitative trait loci (QTL) for resistance to different Pt races from 185 globally diverse wheat genotypes using a genome-wide association study (GWAS) approach. The virulence pattern of the 33 Pt isolates from various wheat-growing areas of Iran on 55 wheat differentials showed that the FKTPS and FKTTS were relatively frequent pathotypes among the 18 identified races. The weighted average frequency of virulence on the resistance genes Lrb, Lr3bg, Lr14b, Lr16, Lr24, Lr3ka, Lr11 and Lr20 were high (> 90%). However, low virulence on the resistant genes Lr2a, Lr9, Lr19, Lr25, Lr28 and Lr29 indicates that these genes are still effective against the pathogen population in Iran at present. GWAS on a panel of 185 wheat genotypes against 10 Pt races resulted into 62 significant marker-trait associations (MTAs) belonged to 34 quantitative trait loci (QTL) across 16 chromosomes. Among them, 10 QTLs on chromosomes 1A, 1B, 3B, 3D, 4A, 6D, 7A and 7D were identified as potential novel QTLs, of which four QTLs (QLr.iau-3B-2, QLr.iau-7A-2, QLr.iau-7A-3 and QLr.iau-7D-2) are more interesting, as they are associated with resistance to two or more Pt races. The known and novel QTLs associated with different Pt races found here, can be used in future wheat breeding programs to recombine different loci for durable resistance against leaf rust races. [ABSTRACT FROM AUTHOR]

SUMMARY: The advent of the pangenome era has unraveled previously unknown genetic variation existing within diverse crop plants, including rice. This untapped genetic variation is believed to account for a major portion of phenotypic variation existing in crop plants. However, the use of conventional single reference‐guided genotyping often fails to capture a large portion of this genetic variation leading to a reference bias. This makes it difficult to identify and utilize novel population/cultivar‐specific genes for crop improvement. Thus, we developed a Rice Pangenome Genotyping Array (RPGA) harboring probes assaying 80K single‐nucleotide polymorphisms (SNPs) and presence–absence variants spanning the entire 3K rice pangenome. This array provides a simple, user‐friendly and cost‐effective (60–80 USD per sample) solution for rapid pangenome‐based genotyping in rice. The genome‐wide association study (GWAS) conducted using RPGA‐SNP genotyping data of a rice diversity panel detected a total of 42 loci, including previously known as well as novel genomic loci regulating grain size/weight traits in rice. Eight of these identified trait‐associated loci (dispensable loci) could not be detected with conventional single reference genome‐based GWAS. A WD repeat‐containing PROTEIN 12 gene underlying one of such dispensable locus on chromosome 7 (qLWR7) along with other non‐dispensable loci were subsequently detected using high‐resolution quantitative trait loci mapping confirming authenticity of RPGA‐led GWAS. This demonstrates the potential of RPGA‐based genotyping to overcome reference bias. The application of RPGA‐based genotyping for population structure analysis, hybridity testing, ultra‐high‐density genetic map construction and chromosome‐level genome assembly, and marker‐assisted selection was also demonstrated. A web application (http://www.rpgaweb.com) was further developed to provide an easy to use platform for the imputation of RPGA‐based genotyping data using 3K rice reference panel and subsequent GWAS. Significance Statement: RPGA is a vital tool for diverse genomics‐assisted breeding applications and accelerated crop improvement in rice, as it provides an efficient pangenome‐based genotyping solution to overcome reference bias imposed by conventional single reference genome‐based genotyping strategies. [ABSTRACT FROM AUTHOR]

The sorghum-sudangrass hybrid is a vital annual gramineous herbage. Few reports exist on its ultra-high-density genetic map. In this study, we sought to create an ultra-high-density genetic linkage map for this hybrid to strengthen its functional genomics research and genetic breeding. We used 150 sorghum-sudangrass hybrid F2 individuals and their parents (scattered ear sorghum and red hull sudangrass) for high-throughput sequencing on the basis of whole genome resequencing. In total, 1,180.66 Gb of data were collected. After identification, filtration for integrity, and partial segregation, over 5,656 single nucleotide polymorphism markers of high quality were detected. An ultra-high-density genetic linkage map was constructed using these data. The markers covered approximately 2,192.84 cM of the map with average marker intervals of 0.39 cM. The length ranged from 115.39 cM to 264.04 cM for the 10 linkage groups. Currently, this represents the first genetic linkage map of this size, number of molecular markers, density, and coverage for sorghum-sudangrass hybrid. The findings of this study provide valuable genome-level information on species evolution and comparative genomics analysis and lay the foundation for further research on quantitative trait loci fine mapping and gene cloning and marker-assisted breeding of important traits in sorghum-sudangrass hybrids. [ABSTRACT FROM AUTHOR]

Stripe rust disease of wheat, caused by Puccinia striiformis f. sp. tritici, (Pst) is one of the most serious diseases of wheat worldwide. In India, virulent stripe rust races have been constantly evolving in the North-Western Plains Zone leading to the failure of some of the most widely grown resistant varieties in the region. With the goal of studying the recent evolution of virulent races in this region, we conducted whole-genome re-sequencing of three prevalent Indian Pst pathotypes Pst46S119, Pst78S84 and Pst110S119. We assembled 58.62, 58.33 and 55.78 Mb of Pst110S119, Pst46S119 and Pst78S84 genome, respectively and found that pathotypes were highly heterozygous. Comparative phylogenetic analysis indicated the recent evolution of pathotypes Pst110S119 and Pst78S84 from Pst46S119. Pathogenicity-related genes classes (CAZyme, proteases, effectors, and secretome proteins) were identified and found to be under positive selection. Higher rate of gene families expansion were also observed in the three pathotypes. A strong association between the effector genes and transposable elements may be the source of the rapid evolution of these strains. Phylogenetic analysis differentiated the Indian races in this study from other known United States, European, African, and Asian races. Diagnostic markers developed for the identification of three Pst pathotypes will help tracking of yellow rust at farmers field and strategizing resistance gene deployment. [ABSTRACT FROM AUTHOR]

Background: For genetic analyses, multi-allelic markers have an advantage over bi-allelic markers like SNPs (single nucleotide polymorphisms) in that they carry more information about the genetic constitution of individuals. This is especially the case in polyploids, where individuals carry more than two alleles at each locus. Haploblocks are multi-allelic markers that can be derived by phasing sets of closely-linked SNP markers. Phased haploblocks, similarly to other multi-allelic markers, will therefore be advantageous in genetic tasks like linkage mapping, QTL mapping and genome-wide association studies. Results: We present a new method to reconstruct haplotypes from SNP dosages derived from genotyping arrays, which is applicable to polyploids. This method is implemented in the software package PolyHaplotyper. In contrast to existing packages for polyploids it makes use of full-sib families among the samples to guide the haplotyping process. We show that in this situation it is much more accurate than other available software, using experimental hexaploid data and simulated tetraploid data. Conclusions: Our method and the software package PolyHaplotyper in which it is implemented extend the available tools for haplotyping in polyploids. They perform especially well in situations where one or more full-sib families are present. [ABSTRACT FROM AUTHOR]

Meagre (Argyrosomus regius), is a benthopelagic species rapidly emerging in aquaculture, due to its low food to biomass conversion rate, good fillet yield and ease of production. Tracing a species genomic background along with describing the genetic basis of important traits can greatly influence both conservation strategies and production perspectives. In this study, we employed ddRAD sequencing of 266 fish from six F1 meagre families, to construct a high-density genetic map comprising 4529 polymorphic SNP markers. The QTL mapping analysis provided a genomic appreciation for the weight trait identifying a statistically significant QTL on linkage group 15 (LG15). The comparative genomics analysis with six teleost species revealed an evolutionarily conserved karyotype structure. The synteny observed, verified the already well-known fusion events of the three-spine stickleback genome, reinforced the evidence of reduced evolutionary distance of Sciaenids with the Sparidae family, reflected the evolutionary proximity with Dicentrarchus labrax, traced several putative chromosomal rearrangements and a prominent putative fusion event in meagre's LG17. This study presents novel elements concerning the genome evolutionary history of a non-model teleost species recently adopted in aquaculture, starts to unravel the genetic basis of the species growth-related traits, and provides a high-density genetic map as a tool that can help to further establish meagre as a valuable resource for research and production. [ABSTRACT FROM AUTHOR]

Shell color shows broad variation within mollusc species and despite information on the genetic pathways involved in shell construction and color has recently increased, more studies are needed to understand its genetic architecture. The common cockle (Cerastoderma edule) is a valuable species from ecological and commercial perspectives which shows important variation in shell color across Northeast Atlantic. In this study, we constructed a high-density genetic map, as a tool for screening common cockle genome, which was applied to ascertain the genetic basis of color variation in the species. The consensus genetic map comprised 19 linkage groups (LGs) in accordance with the cockle karyotype (2n = 38) and spanned 1073 cM, including 730 markers per LG and an inter-marker distance of 0.13 cM. Five full-sib families showing segregation for several color-associated traits were used for a genome-wide association study and a major QTL on chromosome 13 associated to different color-traits was detected. Mining on this genomic region revealed several candidate genes related to shell construction and color. A genomic region previously reported associated with divergent selection in cockle distribution overlapped with this QTL suggesting its putative role on adaptation. [ABSTRACT FROM AUTHOR]

Key message: Rht22 was fine mapped in the interval of 0.53–1.48 Mb on 7AS, which reduces cell number of internode to cause semi-dwarfism in Jianyangailanmai. As a valuable germplasm resource for wheat genetic improvement, tetraploid wheat has several reduced height (Rht) and enhanced harvest index genes. Rht22, discovered in Jianyangailanmai (JAM, Triticum turgidum L., 2n = 4x = 28, AABB), significantly increases the spikelet number per spike, but its accurate chromosomal position is still unknown. In this study, a high-density genetic map was constructed using specific-length amplified fragment sequencing in an F7 RIL_DJ population, which was derived from a cross between dwarf Polish wheat (T. polonicum L., 2n = 4x = 28, AABB) and JAM. Two plant height loci, Qph.sicau-4B and Qph.sicau-7A, were mapped on chromosomes 4BS and 7AS, respectively. Qph.sicau-7A was mapped to the 0.33-4.46 Mb interval on 7AS and likely represents the candidate region of Rht22. Fine mapping confirmed and narrowed Rht22 on chromosome arm 7AS between Xbag295.s53 and Xb295.191 in three different populations. The physical region ranged from 0.53 to 1.48 Mb and included 18 candidate genes. Transcriptome analysis of two pairs of near-isogenic lines revealed that 135 differentially expressed genes (DEGs) were associated with semi-dwarfism. Of these, the expression of 83 annotated DEGs involved in hormones synthesis and signal transduction, cell wall composition, DNA replication, microtubule and phragmoplast arrays was significantly down-regulated in the semi-dwarf line. Therefore, Rht22 causes semi-dwarfism in JAM by disrupting these cellular processes, which impairs cell proliferation and reduces internode cell number. [ABSTRACT FROM AUTHOR]

Key message: A stripe rust resistance gene YrZM175 in Chinese wheat cultivar Zhongmai 175 was mapped to a genomic interval of 636.4 kb on chromosome arm 2AL, and a candidate gene was predicted. Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is a worldwide wheat disease that causes large losses in production. Fine mapping and cloning of resistance genes are important for accurate marker-assisted breeding. Here, we report the fine mapping and candidate gene analysis of stripe rust resistance gene YrZM175 in a Chinese wheat cultivar Zhongmai 175. Fifteen F1, 7,325 F2 plants and 117 F2:3 lines derived from cross Avocet S/Zhongmai 175 were inoculated with PST race CYR32 at the seedling stage in a greenhouse, and F2:3 lines were also evaluated for stripe rust reaction in the field using mixed PST races. Bulked segregant RNA-seq (BSR-seq) analyses revealed 13 SNPs in the region 762.50–768.52 Mb on chromosome arm 2AL. By genome mining, we identified SNPs and InDels between the parents and contrasting bulks and mapped YrZM175 to a 0.72-cM, 636.4-kb interval spanned by YrZM175-InD1 and YrZM175-InD2 (763,452,916–764,089,317 bp) including two putative disease resistance genes based on IWGSC RefSeq v1.0. Collinearity analysis indicated similar target genomic intervals in Chinese Spring, Aegilops tauschii (2D: 647.7–650.5 Mb), Triticum urartu (2A: 750.7–752.3 Mb), Triticum dicoccoides (2A: 771.0–774.5 Mb), Triticum turgidum (2B: 784.7–788.2 Mb), and Triticum aestivum cv. Aikang 58 (2A: 776.3–778.9 Mb) and Jagger (2A: 789.3–791.7 Mb). Through collinearity analysis, sequence alignments of resistant and susceptible parents and gene expression level analysis, we predicted TRITD2Bv1G264480 from Triticum turgidum to be a candidate gene for map-based cloning of YrZM175. A gene-specific marker for TRITD2Bv1G264480 co-segregated with the resistance gene. Molecular marker analysis and stripe rust response data revealed that YrZM175 was different from genes Yr1, Yr17, Yr32, and YrJ22 located on chromosome 2A. Fine mapping of YrZM175 lays a solid foundation for functional gene analysis and marker-assisted selection for improved stripe rust resistance in wheat. [ABSTRACT FROM AUTHOR]

Soybeans are essential crops that supply protein and oil. The composition and contents of soybean fatty acids are relevant to human health and have a significant relationship with soybean oil processing and applications. Identifying quantitative trait locus (QTL) genes related to palmitic acid could facilitate the development of a range of nutritive soybean cultivars using molecular marker-assisted selection. In this study, we used a cultivar with higher palmitic acid content, 'Dongnong42', and a lower palmitic acid content cultivar, 'Hobbit', to establish F2:6 recombinant inbred lines. A high-density genetic map containing 9980 SLAF markers was constructed and distributed across 20 soybean chromosomes. The genetic map contained a total genetic distance of 2602.58 cM and an average genetic distance of 0.39 cM between adjacent markers. Two QTLs related to palmitic acid content were mapped using inclusive composite interval mapping, explaining 4.2–10.1% of the phenotypic variance in three different years and environments, including the QTL included in seed palmitic 7-3, which was validated by developing SSR markers. Based on the SNP/Indel and significant differential expression analyses of Dongnong42 and Hobbit, two genes, Glyma.15g119700 and Glyma.15g119800, were selected as candidate genes. The high-density genetic map, QTLs, and molecular markers will be helpful for the map-based cloning of palmitic acid content genes. These could be used to accelerate breeding for high nutritive value cultivars via molecular marker-assisted breeding. [ABSTRACT FROM AUTHOR]

With global warming and regional decreases in precipitation, drought has become a problem worldwide. As the number of arid regions in the world is increasing, drought has become a major factor leading to significant crop yield reductions and food crises. Soybean is a crop that is relatively sensitive to drought. It is also a crop that requires more water during growth and development. The aim of this study was to identify the quantitative trait locus (QTL) that affects drought tolerance in soybean by using a recombinant inbred line (RIL) population from a cross between the drought-tolerant cultivar 'Jindou21' and the drought-sensitive cultivar 'Zhongdou33'. Nine agronomic and physiological traits were identified under drought and well-watered conditions. Genetic maps were constructed with 923,420 polymorphic single nucleotide polymorphism (SNP) markers distributed on 20 chromosomes at an average genetic distance of 0.57 centimorgan (cM) between markers. A total of five QTLs with a logarithm of odds (LOD) value of 4.035–8.681 were identified on five chromosomes. Under well-watered conditions and drought-stress conditions, one QTL related to the main stem node number was located on chromosome 16, accounting for 17.177% of the phenotypic variation. Nine candidate genes for drought resistance were screened from this QTL, namely Glyma.16G036700, Glyma.16G036400, Glyma.16G036600, Glyma.16G036800, Glyma.13G312700, Glyma.13G312800, Glyma.16G042900, Glyma.16G043200, and Glyma.15G100700. These genes were annotated as NAC transport factor, GATA transport factor, and BTB/POZ-MATH proteins. This result can be used for molecular marker-assisted selection and provide a reference for breeding for drought tolerance in soybean. [ABSTRACT FROM AUTHOR]

Fruit shape is one of the important quantitative traits in pear (Pyrus spp.) breeding program, thus genetic study related to fruit shape could be beneficial to pear breeding. Quantitative trait loci (QTL) analysis was carried out using 'Whangkeumbae' (P. pyrifolia, round) × 'Yali' (P. bretschneideri, pyriform) and high-resolution melting (HRM) markers were developed. The genetic linkage map of 'Whangkeumbae' × 'Yali' was constructed using single nucleotide polymorphisms (SNPs) derived from Axiom Pear 70 K Genotyping Array and simple sequence repeats. The integrated genetic linkage map of 'Whangkeumbae' × 'Yali' showed ~ 90% of genome coverage, a total genetic distance of 998.2 cM, and a marker density of 1.6 cM. F1 progenies of 'Whangkeumbae' × 'Yali' showed normal distribution of fruit length (L), diameter (D), and L/D ratio. Three QTLs located in linkage group (LG) 6, 7, and 12 were identified with LOD thresholds of 2.8–3.0. Six HRM markers were developed using array-SNPs anchored in the QTLs and predicted fruit shape with 28.6–65.3% accuracy. Notably, accuracy was increased by ~ 90% using an HRM marker combination consisting of CBp06sn02, CBp07sn01, and CBp12sn03. These results could provide a better understanding of the genetic mechanism of fruit shape development and reducing pear breeding period. [ABSTRACT FROM AUTHOR]

Melon (Cucumis melo L.) production is often restricted by a plethora of pests and diseases, including powdery mildew and downy mildew caused respectively by the fungal species Podosphaera xanthii/Golovinomyces orontii and oomycete species Pseudoperonospora cubensis. Many efforts have been directed on identification of resistant sources by screening (wild) melon germplasm. In the current review, we summarized such efforts from various publications of the last 50 plus years. Resistance to powdery mildew has been identified in 239 melon accessions and downy mildew resistance in 452 accessions of both C. melo and the wild relative species C. figarei. Among the resistance sources, C. melo var. cantalupensis accessions PMR 45, PMR 5, PMR 6, and WMR 29 as well as C. melo var. momordica accessions PI 124111, PI 124112, and PI 414723 have been considered as the most valuable germplasm because multiple resistance genes have been identified from these accessions and are widely used in melon resistance breeding. Further genetic mapping in a number of resistant sources has enabled identification of 25 dominant genes, two recessive genes and seven QTLs conferring powdery mildew resistance, as well as eight dominant genes and 11 QTLs for downy mildew resistances. Based on the reported sequences of associated markers, we anchored physically (many of) these genes and QTLs to chromosomes of the melon cv. DHL92 genome. In addition to presenting a comprehensive overview on powdery mildew and downy mildew resistance in (wild) melon germplasm, we suggest strategies aiming at breeding melon with durable and broad-spectrum resistance to pathogens and pests [ABSTRACT FROM AUTHOR]

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 F2 individuals generated from PPIS and PPS. In total, 4 655 SLAFs were selected and mapped on 20 linkage groups (LGs). The total map length was 2 502.01 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 F2 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. [ABSTRACT FROM AUTHOR]

Durian (Durio zibethinus L.) is a crop of economic and health importance globally. Efforts are being made to revamp China's only successful commercial-scale durian plantations in Hainan; however, their genetic base is unknown. Therefore, the present study was undertaken to assess the genetic base and population structure of 32 genotypes in durian plantation sites in Hainan, China, and develop simple sequence repeat (SSR) markers by whole genome sequencing through restriction site-associated DNA sequencing technology to facilitate germplasm conservation and breeding. The results from identity by state (IBS), phylogenetic tree, population structure, and principal component analysis grouped the 32 genotypes into two clusters/sub-populations. Based on IBS, genotypes in Cluster I are largely duplicated genotypes; however, results from the model-based population structure demonstrated that most of the genotypes in Sub-population II shared a common genetic background with those in Sub-population I/Cluster I. The results revealed that the core durian collection in the plantation sites in Hainan include D24, D101, MSW, JH, D163, HFH, and NLX-5. In addition, we developed a total of 79,178 SSR markers with varied lengths and amplicon sizes. The genetic diversity and population structure reported in this study will be useful for durian conservation and utilization. In addition, the discovered and developed SSR markers will lay the foundation for molecular breeding via marker-assisted selection, quantitative trait loci mapping, and candidate gene discovery and validation. [ABSTRACT FROM AUTHOR]

Rice as one of the important crops, is limited by salt stress in different growth stages. To identify genomic regions associated with morphological traits in seedling, vegetative and reproductive stages of rice in salt stress, three experiments were conducted as completely randomized design using an F8 RIL population caused two Indica varieties Tarom Mahali (salt-tolerant) and Khazar (salt-sensitive) cross. The F8 lines were planted in non-stress and NaCl stress (8 dS m–1) conditions. A linkage map was constructed by SSR, ISSR, and IRAP markers. The genetic map contained 12 linkage groups that covered 1042.2 cM with a 12.86 cM distance between two markers on average. qNRFW-1b QTL was identified as major QTLs (LOD = 4.02 and R2 = 15.00) for root fresh weight under non-stress condition, at seedling stage. In vegetative stage, qSPH120-9, qPH120-10 and qSSDW-1 were identified as major effect QTLs associated with plant height and shoot dry weight under salt stress condition. In reproductive stage, qNPH-1, qNRDW-1b (under non-stress condition), qSMP-1d and qSSSC-2 (under salt stress condition), were major QTLs for Plant height and stem dry weight, maturation period and stem sodium content, which explained 18, 25.4, 15.9 and 18.7% of the total phenotypic variation. QTLs with pleiotropic effects were identified in both conditions and all three stages which might be for multiple QTL introgression for marker-assisted breeding programs to improve the salinity tolerance. After validation, the major QTLs identified in the present study can be considered for use in marker-assisted selection (MAS) programs. [ABSTRACT FROM AUTHOR]

Common buckwheat (Fagopyrum esculentum M.) is an important pseudo-cereal crop and contains an abundance of nutrients and bioactive compounds. However, the yield of buckwheat is low compared to that of other major crops. QTL mapping and candidate gene screening of grain size are very important for increasing production in common buckwheat through molecular breeding in the future. In the present study, an F1 segregating population with 217 individuals was established using a cross between Ukraine daliqiao (UD) and Youqiao 2 (YQ2) that showed a significant difference in grain size. The InDel and SSR primers were developed based on transcriptome sequencing between parents in the previous study. We constructed a genetic linkage map, including 39 SSR loci and 93 InDel loci, with a total length of 1398.33 cM and an average spacing of 10.59 cM. Combined with the grain size phenotype data of the F1 population, a total of 14 QTL were detected, including 6-grain length QTL, 3-grain width QTL, and 5 hundred-grain-weight QTL. QTL of grain width and hundred-grain weight were all detected near SWU_Fe_InDel086 and SWU_Fe_InDel076. Some putative candidate genes with the ex1pression or InDel difference between parents were screened within the QTL interval. This study would lay the foundation for map-based cloning and molecular mechanism of grain size and ultimately improvement of yield in common buckwheat. [ABSTRACT FROM AUTHOR]

Wax gourd (Benincasa hispida [Thunb] Cogn.) is an important horticultural crop in the Cucurbitaceae family that has high nutritive and medicinal value. We analysed quantitative trait loci (QTLs) related to the fruit-related traits of wax gourd by constructing a high-density genetic map through whole-genome resequencing. We obtained an F2 population of 195 individuals by crossing two inbred wax gourd lines: XDJQ-1 and SL-7. A high-density genetic map was constructed after generating a sequencing library and identifying and genotyping single-nucleotide polymorphisms (SNPs). Finally, we used QTL mapping to analyse the fruit-related traits of wax gourd. The high-density genetic map showed sufficient coverage to detect recombination breakpoints and contained 323,107 SNP markers, spanned 1243.87 cM, and had an average distance between adjacent markers of 0.33 cM. Furthermore, 3,720 bin markers (including SNP markers) were mapped to 12 linkage groups. Based on the high-density genetic map, one major QTL for fruit flesh colour (fc5.1) was mapped on the 3.379 Mb region of chromosome 5, and two major QTLs for fruit length (fl2.1) and fruit width (fw2.1) were mapped on the (0.345 Mb) and (0.602 Mb) regions of chromosome 2 respectively. The QTLs for fc5.1, fl2.1, and fw2.1 accounted for 28.684%, 63.562%, and 59.979% of wax gourd phenotypic variation, respectively. To our knowledge, ours is the first high-density genetic map of wax gourd constructed using whole-genome resequencing and is the highest density genetic map available for wax gourd. Our results provide a foundation for future map-based cloning, candidate gene identification, and marker-assisted breeding. [ABSTRACT FROM AUTHOR]

Pumpkin is a popular vegetable crop and exhibits a broad diversity in fruit shape and size. Fruit-related traits are the decisive factors determining consumer acceptance and market value of pumpkin cultivar. As a result, deciphering the genetic basis of fruit-related traits is of great importance for pumpkin breeding. To address this problem, a F2 population was generated by two Cucurbita moschata inbred lines with contrasting fruit shapes, and genotyping by sequencing approach was used to construct a genetic map and localize the QTLs underlying the fruit-related traits in this study. The results showed that a high-quality genetic map was constructed for pumpkin, which comprised of 2413 bins and spanned a total length of 2252.10 cM with an average genetic distance of 0.94 cM. Thirty significant QTLs with moderate or small effects were identified for seven fruit-related traits, including fruit length, fruit diameter, fruit shape index, fruit weight, fruit flesh thickness, seed cavity size, and total soluble solids content. Co-locations were observed between the QTLs underlying different traits, demonstrating that pleiotropic effect plays an important role in genetic control of fruit-related traits. The identified QTLs provide valuable information for further fine mapping of the related genes and pumpkin breeding programs with the aim of improving fruit quality. [ABSTRACT FROM AUTHOR]

Genetic linkage maps are used to localize markers on the genome based on the recombination frequency. Most often, these maps are based on the segregation observed within a single biparental population of limited size (n < 300) where relatively few recombination events are sampled and in which some genomic regions are monomorphic because both parents carry the same alleles. Together, these two limitations affect both the resolution and extent of genome coverage of such maps. Consensus genetic maps overcome the limitations of individual genetic maps by merging the information from multiple segregating populations derived from a greater diversity of parental combinations, thus increasing the number of recombination events and reducing the number of monomorphic regions. The aim of this study was to construct a high-density consensus genetic map for single nucleotide polymorphism (SNP) markers obtained through a genotyping-by-sequencing (GBS) approach. Individual genetic maps were generated from six F4:5 mapping populations (n = 278–365), totaling 1857 individuals. The six linkage maps were then merged to produce a consensus map comprising a total of 16 311 mapped SNPs that jointly cover 99.5% of the soybean genome with only two gaps larger than 10 cM. Compared to previous soybean consensus maps, it offers a more extensive and uniform coverage. [ABSTRACT FROM AUTHOR]

Two sudangrass varieties, Sa and S722, were sequenced using restriction site-associated DNA sequencing (RAD-seq) on the Illumina Miniseq platform. After quality control, 920 542 and 892 626 clean reads were obtained for Sa and S722, respectively. Compared with the sorghum reference genome, 2341 and 2123 single nucleotide polymorphisms (SNPs) were obtained from Sa and S722, respectively. A total of 543 and 472 insertion-deletion (InDel) loci were obtained by sequence analysis in Sa and S722, respectively. From these InDel loci, 100 InDels were randomly selected to design InDel markers from Sa and S722. Polymorphism analyses were performed between sorghum Tx623B and Sa and between Tx623B and S722 using the InDel markers. The results showed that the polymorphism between Tx623B and Sa was 85%, and that between Tx623B and S722 was 87%. Diversity analysis was performed using 39 InDel markers for 42 sorghum and 6 sudangrass germplasms. Statistical analyses showed that the Shannon information index was 0.10–1.09 with an average value of 0.54. The polymorphism information content of the InDel markers ranged from 0.04 to 0.66 with an average of 0.35. Two genetic maps of chromosomes 1 and 2 of the Wancao No. 2 recombinant inbred line population were constructed using 16 InDel markers. Thus, it is an effective method to develop InDel markers for sorghum and sudangrass using sudangrass RAD-seq data. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Sugarcane leaf blight (SLB), caused by Stagonospora tainanensis, is one of the most harmful fungal diseases, threatening the sugarcane industry and causing high losses of cane yield and sugar in susceptible cultivars. Using a two-way pseudo-testcross mapping strategy in combination with array genotyping, two high-density genetic maps were constructed for sugarcane cultivars YT93-159 and ROC22 with mean densities of respectively 3.0 and 3.5 cM per marker, and covering respectively 4485 and 2720 cM of genetic distance. The maps showed highly conserved colinearity with the genome of the ancestral species Saccharum officinarum, supporting the reliability of the linkage configurations of the maps. Quantitative trait loci (QTL) analysis of SLB resistance revealed six QTL (qSLB-1-qSLB-6). The major QTL qSLB-1 explaining 16.4% of phenotypic variance was assigned as the main QTL, and the total percentages of phenotypic variance explained in YT93-159 and ROC22 were 37.9% and 17.6%, respectively. Nine transcription factor and seven pathogen receptor genes lying in the qSLB-1 interval were highly expressed and are proposed as candidate causal genes for SLB resistance. [ABSTRACT FROM AUTHOR]

Powdery mildew (PM), caused by the fungus Microsphaera diffusa, causes severe yield losses in soybean [Glycine max (L.) Merr.] under suitable environmental conditions. Identifying resistance genes and developing resistant cultivars may prevent soybean PM damage. In this study, analysis of F1, F2, and F8:11 recombinant inbred line (RIL) populations derived from the cross between Zhonghuang 24 (ZH24) and Huaxia 3 (HX3) indicated that adult-plant resistance (APR) to powdery mildew in the soybean cultivar (cv.) ZH24 was controlled by a single dominant locus. A high-density genetic linkage map of the RIL population was used for fine mapping. The APR locus in ZH24 was mapped to a 281-kb genomic region on chromosome 16. Using 283 susceptible plants of another F2 population, the candidate region was finemapped to a 32.8-kb genomic interval flanked by the markers InDel14 and Gm16_428. The interval harbored five genes, including four disease resistance (R)-like genes, according to the Williams 82.a2.v1 reference genome. Quantitative real-time PCR assays of candidate genes revealed that the expression levels of Glyma.16g214300 and Glyma.16g214500 were changed by M. diffusa infection and might be involved in disease defense. Rmd_B13 showed all-stage resistance (ASR) to PM in soybean cv. B13. An allelism test in the F2 segregating population from the cross of ZH24 × B13 suggested that the APR locus Rmd_ZH24 and the ASR locus Rmd_B13 may be allelic or tightly linked. These results provide a reference marker-assisted selection in breeding programs. [ABSTRACT FROM AUTHOR]

Leaf development underlies crop growth and productivity and has been a major target of crop domestication and improvement. However, most genes controlling leaf development in barley remain unknown. We identified a dwarf and liguleless (dl) mutant derived by ethylmethane sulfonate mutagenesis. The dl mutant showed dramatic changes in shoot architecture compared with wild-type (Yangnongpi 5) plants. Besides lacking ligules, the dl mutant showed much shorter plant height (28 cm) than Yangnongpi 5 (78 cm). By map-based cloning, the dl gene was localized to a 56.58-kb genomic interval on the long arm of chromosome 7. A C-to-T single-nucleotide substitution was identified at exon position 790, and is a functional mutation resulting in a proline-to-serine substitution at the 264th amino acid residue of HORVU7Hr1G106960. Consequently, HORVU7Hr1G106960 was identified as the DL gene, encoding 269 amino acids and containing the Arabidopsis LSH1 and Oryza G1 (ALOG) domain. DL is highly similar to rice OsG1-LIKE 1/2 (OsG1L1/2) and sorghum AWN1/AWN1-10 at the amino acid level. Although the dl mutant allele showed no expression changes in selected tissues by real-time PCR, we propose HORVU7Hr1G106960 as a candidate gene conferring the dwarf and liguleless phenotype in barley. [ABSTRACT FROM AUTHOR]

Background: Flowering time is an important agronomic trait of crops and significantly affects plant adaptation and seed production. Flowering time varies greatly among maize (Zea mays) inbred lines, but the genetic basis of this variation is not well understood. Here, we report the comprehensive genetic architecture of six flowering time-related traits using a recombinant inbred line (RIL) population obtained from a cross between two maize genotypes, B73 and Abe2, and combined with genome-wide association studies to identify candidate genes that affect flowering time. Results: Our results indicate that these six traits showed extensive phenotypic variation and high heritability in the RIL population. The flowering time of this RIL population showed little correlation with the leaf number under different environmental conditions. A genetic linkage map was constructed by 10,114 polymorphic markers covering the whole maize genome, which was applied to QTL mapping for these traits, and identified a total of 82 QTLs that contain 13 flowering genes. Furthermore, a combined genome-wide association study and linkage mapping analysis revealed 17 new candidate genes associated with flowering time. Conclusions: In the present study, by using genetic mapping and GWAS approaches with the RIL population, we revealed a list of genomic regions and candidate genes that were significantly associated with flowering time. This work provides an important resource for the breeding of flowering time traits in maize. [ABSTRACT FROM AUTHOR]

Background: Flowering time is an important agronomic trait of crops and significantly affects plant adaptation and seed production. Flowering time varies greatly among maize (Zea mays) inbred lines, but the genetic basis of this variation is not well understood. Here, we report the comprehensive genetic architecture of six flowering time-related traits using a recombinant inbred line (RIL) population obtained from a cross between two maize genotypes, B73 and Abe2, and combined with genome-wide association studies to identify candidate genes that affect flowering time. Results: Our results indicate that these six traits showed extensive phenotypic variation and high heritability in the RIL population. The flowering time of this RIL population showed little correlation with the leaf number under different environmental conditions. A genetic linkage map was constructed by 10,114 polymorphic markers covering the whole maize genome, which was applied to QTL mapping for these traits, and identified a total of 82 QTLs that contain 13 flowering genes. Furthermore, a combined genome-wide association study and linkage mapping analysis revealed 17 new candidate genes associated with flowering time. Conclusions: In the present study, by using genetic mapping and GWAS approaches with the RIL population, we revealed a list of genomic regions and candidate genes that were significantly associated with flowering time. This work provides an important resource for the breeding of flowering time traits in maize. [ABSTRACT FROM AUTHOR]

Key message: qFT7.1, a major QTL for flowering time in Brassica rapa was fine-mapped to chromosome A07 in a 56.4-kb interval, in which the most likely candidate gene is BraA07g018240.3C. In Brassica rapa, flowering time (FT) is an important agronomic trait that affects the yield, quality, and adaption. FT is a complicated trait that is regulated by many genes and is affected greatly by the environment. In this study, a chromosome segment substitution line (CSSL), CSSL16, was selected that showed later flowering than the recurrent parent, a rapid-cycling inbred line of B. rapa (RcBr). Using Bulked Segregant RNA sequencing, we identified a late flowering quantitative trait locus (QTL), designated as qFT7.1, on chromosome A07, based on a secondary-F2 population derived from the cross between CSSL16 and RcBr. qFT7.1 was further validated by conventional QTL mapping. This QTL explained 39.9% (logarithm of odds = 32.2) of the phenotypic variations and was fine mapped to a 56.4-kb interval using recombinant analysis. Expression analysis suggested that BraA07g018240.3C, which is homologous to ATC (encoding Arabidopsis thaliana CENTRORADIALIS homologue), a gene for delayed flowering in Arabidopsis, as the most promising candidate gene. Sequence analysis demonstrated that two synonymous mutations existed in the coding region and numerous bases replacements existed in promoter region between BraA07g018240.3C from CSSL16 and RcBr. The results will increase our knowledge related to the molecular mechanism of late flowering in B. rapa and lays a solid foundation for the breeding of late bolting B. rapa. [ABSTRACT FROM AUTHOR]

Key message: A powerful QTL analysis method for nested association mapping populations is presented. Based on a one-stage multi-locus model, it provides accurate predictions of founder specific QTL effects. Nested association mapping (NAM) populations have been created to enable the identification of quantitative trait loci (QTL) in different genetic backgrounds. A whole-genome nested association mapping (WGNAM) method is presented to perform QTL analysis in NAM populations. The WGNAM method is an adaptation of the multi-parent whole genome average interval mapping approach where the crossing design is incorporated through the probability of inheriting founder alleles for every marker across the genome. Based on a linear mixed model, this method provides a one-stage analysis of raw phenotypic data, molecular markers, and crossing design. It simultaneously scans the whole-genome through an iterative process leading to a model with all the identified QTL while keeping the false positive rate low. The WGNAM approach was assessed through a simulation study, confirming to be a powerful and accurate method for QTL analysis for a NAM population. This novel method can also accommodate a multi-reference NAM (MR-NAM) population where donor parents are crossed with multiple reference parents to increase genetic diversity. Therefore, a demonstration is presented using a MR-NAM population for wheat (Triticum aestivum L.) to perform a QTL analysis for plant height. The strength and size of the putative QTL were summarized enhancing the understanding of the QTL effects depending on the parental origin. Compared to other methods, the proposed methodology based on a one-stage analysis provides greater power to detect QTL and increased accuracy in the estimation of their effects. The WGNAM method establishes the basis for accurate QTL mapping studies for NAM and MR-NAM populations. [ABSTRACT FROM AUTHOR]

As a desirable agricultural trait, multi-inflorescence (MI) fulfills the requirement of mechanized harvesting and yield increase in rapeseed (Brassica napus L.). However, the genetic mechanism underlying the multi-inflorescence trait remain poorly understood. We previously identified a difference of one pair of dominant genes between the two mapping parental materials. In this study, phenotype and expression analysis indicated that the imbalance of the CLAVATA (CLV)-WUSCHEL (WUS) feedback loop may contribute to the abnormal development of the shoot apical meristem (SAM). BnaMI was fine-mapped to a 55 kb genomic region combining with genotype and phenotype of 5768 BCF1 individuals using a traditional mapping approach. Through comparative and expression analyses, combined with the annotation in Arabidopsis, five genes in this interval were identified as candidate genes. The present findings may provide assistance in functional analysis of the mechanism associated with multi-inflorescence and yield increase in rapeseed. [ABSTRACT FROM AUTHOR]

Sexual reproduction involves meiotic recombination and the creation of crossing over between homologous chromosomes, which leads to new allele combinations. We present a new approach that uses the allele frequency differences and the physical distance of neighboring polymorphisms to estimate the recombination rate from pool genotyping or sequencing. This allows a considerable cost reduction compared to conventional mapping based on genotyping or sequencing data of single individuals. We evaluated the approach based on computer simulations at various genotyping depths and population sizes as well as applied it to experimental data of 45 barley populations, comprising 4182 RIL. High correlations between the recombination rates from this new pool genetic mapping approach and conventional mapping in simulated and experimental barley populations were observed. The proposed method therefore provides a reliable genetic map position and recombination rate estimation in defined genomic windows. [ABSTRACT FROM AUTHOR]