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Safflower oil with high oleic is an important source of oil due to its oxidative stability and nutritional value as well as its industrial benefits. Identification of molecular markers associated with oil quality traits may aid in genotype selection in breeding efforts of safflower. To determine the genetic basis of seed oil, oleic acid, and linoleic acid content in safflower seeds, a recombinant inbred population was developed from a cross between Goldasht (low oleic line) and Mex 22–191 (high oleic line) by individual self-pollinated plants in each generation (from F2 to F9). Fatty acid components were detected by gas chromatography and subsequently analyzed by QTL analysis. The high variability in fatty acid composition was demonstrated and the first high-oleic safflower genotype was generated. A new linkage map with 69 AFLP and 45 ISSR polymorphic markers was made, covering 1472,441 cM of the safflower genome. A total of 63 individual QTLs and 238 epistatic QTLs, related to oil quality, were detected in 14 genomic regions. Some major QTLs with phenotypic variation greater than 10% were identified for the measured traits. These results demonstrate that major QTL may play an important role in increase the oleic acid of the genetic material used. Furthermore, the larger number of minor QTLs and epistatic QTLs, both of which have additive effects, indicates that the expression of these traits is also influenced by interactions between multiple genes. Our results provide additional genetic resources that can be used to precisely breed these traits in the future. [ABSTRACT FROM AUTHOR]

• This work represents the first-ever detailed multiple marker studies in Stevia. • QTL identification is the novelty of the work that will promote stevia breeding. • The QTL identified could be used in the mapping of genes responsible for rebaudioside-A. • Functional polymorphic markers will be useful in genetic dissection of traits. Stevia rebaudiana Bertoni, belonging to the family Asteraceae, is one of the most important perennial medicinal plants. It is used as a no-calorie sweetener therefore; mining of sweetening genes using molecular markers is a thrust area. In the present study combinations of markers were screened. Out of 170 RAPD, 26 ISSR, and 89 EST-SSR primers, it was observed that 36 RAPD, 10 ISSR, and 33 EST-SSR were found to be polymorphic. Using MAPMAKER/EXP version 3.0 b, four linkage groups were constructed spanning a distance of 927.3 cM with an average distance between loci of 16.29 cM. On QTL identification, a total of 53 locations were found for both trait 1 (rebaudioside-A) and trait 2 (stevioside). Among 53 QTL locations of trait 1, two major QTL were found for rebaudioside-A namely, in the marker interval of L67–L71 (ISSR HB-11)–(IISRS-3-L) in LG1 and the marker interval of L38–L40 (Sigma-5383–027)- (Sigma-5383-029) in LG3 at LOD of 2.5 and 2.7, respectively. The map constructed and QTL identified will greatly facilitate further genetic studies. It is hoped that this work will have a practical impact on the advancement of stevia breeding and genome analysis. [ABSTRACT FROM AUTHOR]

Soybean is a major crop in the world and an essential source for minerals, including iron (Fe) and zin (Zn). Deficiency of Fe and Zn in soil and soybean plants result in yield loss and poor seed nutritional qualities. Information on genomic regions and candidate genes controlling seed Fe and Zn accumulation in soybean seed is limited. Therefore, The objective of this research was to identify genetic regions, known as quantitative trait loci (QTL), and candidate genes that control the accumulation of Fe and Zn in soybean mature seeds. A ‘Forrest’ by ‘Williams 82’ (F × W82) recombinant inbred line (RIL) population (n = 306) was used and genotyped using a total of 5405 single nucleotides polymorphism (SNP) markers using Infinium SNP6K BeadChips. A two-year experiment was conducted across two environments: North Carolina in 2018 (NC) and Illinois in 2020 (IL). Only QTL with LOD scores ≥ 2.5, as identified by the composite interval mapping (CIM) method, are reported here. In total, 6 QTL were identified for seed Fe; specifically, 3 QTL (qFe-01-[NC-2018], qFe-02-[NC-2018], and qFe-03-[NC-2018]) were located on chromosomes 1, 2, and 6, respectively, in the NC environment, and 3 QTL (qFe-01-[IL-2020], qFe-02-[IL-2020], and qFe-03-[IL-2020]) were positioned on chromosomes 1, 2, and 12, respectively, in the IL environment. A total of 6 QTL associated with seed Zn were also identified; 4 QTL (qZn-01-[NC-2018]; qZn-02-[NC-2018]; qZn-03-[NC-2018]; and qZn-04-[NC-2018]), respectively on Chr 2, 3, 7, and 19 in NC; and 2 QTL (qZn-01-[IL-2020] and qZn-02-[IL-2020]), respectively, on Chr 5 and 8 in IL. Several functional genes encode Fe- and Zn-proteins, transcription factors, proteins-zinc finger motifs (involved in DNA binding and transcriptional regulation; crosstalk between the regulatory pathways of Zn and Fe transporters) were identified and located within the QTL interval. To our knowledge, and based on the literature available, the QTL identified here on Chr 2 and Chr 6 are novel and were not previously identified. This current research provides a new knowledge of the genetic basis of seed Fe and Zn and the markers associated with QTL. The QTL identified here will contribute to efficient marker assisted selection for higher Fe and Zn content in soybean seeds. The candidate genes and metal-responsive transcription factors may coordinate the expression of both Zn and Fe transporters in response to changes in metal availability, providing new knowledge on minerals uptake and transport mechanisms, allowing for possible genetic engineering application.

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]

Sheath blight (ShB) is the most serious disease of rice (Oryza sativa L.), caused by the soil-borne fungus Rhizoctonia solani Kühn (R. solani). It poses a significant threat to global rice productivity, resulting in approximately 50% annual yield loss. Managing ShB is particularly challenging due to the broad host range of the pathogen, its necrotrophic nature, the emergence of new races, and the limited availability of highly resistant germplasm. In this study, we conducted QTL mapping using an F2 population derived from a cross between a partially resistant accession (IRGC81941A) of Oryza nivara and the susceptible rice cultivar Punjab rice 121 (PR121). Our analysis identified 29 QTLs for ShB resistance, collectively explaining a phenotypic variance ranging from 4.70 to 48.05%. Notably, a cluster of four QTLs (qRLH1.1, qRLH1.2, qRLH1.5, and qRLH1.8) on chromosome 1 consistently exhibit a resistant response against R. solani. These QTLs span from 0.096 to 420.1 Kb on the rice reference genome and contain several important genes, including Ser/Thr protein kinase, auxin-responsive protein, protease inhibitor/seed storage/LTP family protein, MLO domain-containing protein, disease-responsive protein, thaumatin-like protein, Avr9/Cf9-eliciting protein, and various transcription factors. Additionally, simple sequence repeats (SSR) markers RM212 and RM246 linked to these QTLs effectively distinguish resistant and susceptible rice cultivars, showing great promise for marker-assisted selection programs. Furthermore, our study identified pre-breeding lines in the advanced backcrossed population that exhibited superior agronomic traits and sheath blight resistance compared to the recurrent parent. These promising lines hold significant potential for enhancing the sheath blight resistance in elite cultivars through targeted improvement efforts. [ABSTRACT FROM AUTHOR]

Soybean is a major crop in the world and an essential source for minerals, including iron (Fe) and zin (Zn). Deficiency of Fe and Zn in soil and soybean plants result in yield loss and poor seed nutritional qualities. Information on genomic regions and candidate genes controlling seed Fe and Zn accumulation in soybean seed is limited. Therefore, The objective of this research was to identify genetic regions, known as quantitative trait loci (QTL), and candidate genes that control the accumulation of Fe and Zn in soybean mature seeds. A 'Forrest' by 'Williams 82' (F × W82) recombinant inbred line (RIL) population (n = 306) was used and genotyped using a total of 5405 single nucleotides polymorphism (SNP) markers using Infinium SNP6K BeadChips. A two-year experiment was conducted across two environments: North Carolina in 2018 (NC) and Illinois in 2020 (IL). Only QTL with LOD scores ≥ 2.5, as identified by the composite interval mapping (CIM) method, are reported here. In total, 6 QTL were identified for seed Fe; specifically, 3 QTL (qFe-01-[NC-2018], qFe-02-[NC-2018], and qFe-03-[NC-2018]) were located on chromosomes 1, 2, and 6, respectively, in the NC environment, and 3 QTL (qFe-01-[IL-2020], qFe-02-[IL-2020], and qFe-03-[IL-2020]) were positioned on chromosomes 1, 2, and 12, respectively, in the IL environment. A total of 6 QTL associated with seed Zn were also identified; 4 QTL (qZn-01-[NC-2018]; qZn-02-[NC-2018]; qZn-03-[NC-2018]; and qZn-04-[NC-2018]), respectively on Chr 2, 3, 7, and 19 in NC; and 2 QTL (qZn-01-[IL-2020] and qZn-02-[IL-2020]), respectively, on Chr 5 and 8 in IL. Several functional genes encode Fe- and Zn-proteins, transcription factors, proteins-zinc finger motifs (involved in DNA binding and transcriptional regulation; crosstalk between the regulatory pathways of Zn and Fe transporters) were identified and located within the QTL interval. To our knowledge, and based on the literature available, the QTL identified here on Chr 2 and Chr 6 are novel and were not previously identified. This current research provides a new knowledge of the genetic basis of seed Fe and Zn and the markers associated with QTL. The QTL identified here will contribute to efficient marker assisted selection for higher Fe and Zn content in soybean seeds. The candidate genes and metal-responsive transcription factors may coordinate the expression of both Zn and Fe transporters in response to changes in metal availability, providing new knowledge on minerals uptake and transport mechanisms, allowing for possible genetic engineering application. [ABSTRACT FROM AUTHOR]

Abstract Linkage maps are essential for genetic mapping of phenotypic traits, gene map-based cloning, and marker-assisted selection in breeding applications. Construction of a high-quality saturated map requires high-quality genotypic data on a large number of molecular markers. Errors in genotyping cannot be completely avoided, no matter what platform is used. When genotyping error reaches a threshold level, it will seriously affect the accuracy of the constructed map and the reliability of consequent genetic studies. In this study, repeated genotyping of two recombinant inbred line (RIL) populations derived from crosses Yangxiaomai × Zhongyou 9507 and Jingshuang 16 × Bainong 64 was used to investigate the effect of genotyping errors on linkage map construction. Inconsistent data points between the two replications were regarded as genotyping errors, which were classified into three types. Genotyping errors were treated as missing values, and therefore the non-erroneous data set was generated. Firstly, linkage maps were constructed using the two replicates as well as the non-erroneous data set. Secondly, error correction methods implemented in software packages QTL IciMapping (EC) and Genotype-Corrector (GC) were applied to the two replicates. Linkage maps were therefore constructed based on the corrected genotypes and then compared with those from the non-erroneous data set. Simulation study was performed by considering different levels of genotyping errors to investigate the impact of errors and the accuracy of error correction methods. Results indicated that map length and marker order differed among the two replicates and the non-erroneous data sets in both RIL populations. For both actual and simulated populations, map length was expanded as the increase in error rate, and the correlation coefficient between linkage and physical maps became lower. Map quality can be improved by repeated genotyping and error correction algorithm. When it is impossible to genotype the whole mapping population repeatedly, 30% would be recommended in repeated genotyping. The EC method had a much lower false positive rate than did the GC method under different error rates. This study systematically expounded the impact of genotyping errors on linkage analysis, providing potential guidelines for improving the accuracy of linkage maps in the presence of genotyping errors.

Linkage maps are essential for genetic mapping of phenotypic traits, gene map-based cloning, and marker-assisted selection in breeding applications. Construction of a high-quality saturated map requires high-quality genotypic data on a large number of molecular markers. Errors in genotyping cannot be completely avoided, no matter what platform is used. When genotyping error reaches a threshold level, it will seriously affect the accuracy of the constructed map and the reliability of consequent genetic studies. In this study, repeated genotyping of two recombinant inbred line (RIL) populations derived from crosses Yangxiaomai × Zhongyou 9507 and Jingshuang 16 × Bainong 64 was used to investigate the effect of genotyping errors on linkage map construction. Inconsistent data points between the two replications were regarded as genotyping errors, which were classified into three types. Genotyping errors were treated as missing values, and therefore the non-erroneous data set was generated. Firstly, linkage maps were constructed using the two replicates as well as the non-erroneous data set. Secondly, error correction methods implemented in software packages QTL IciMapping (EC) and Genotype-Corrector (GC) were applied to the two replicates. Linkage maps were therefore constructed based on the corrected genotypes and then compared with those from the non-erroneous data set. Simulation study was performed by considering different levels of genotyping errors to investigate the impact of errors and the accuracy of error correction methods. Results indicated that map length and marker order differed among the two replicates and the non-erroneous data sets in both RIL populations. For both actual and simulated populations, map length was expanded as the increase in error rate, and the correlation coefficient between linkage and physical maps became lower. Map quality can be improved by repeated genotyping and error correction algorithm. When it is impossible to genotype the whole mapping population repeatedly, 30% would be recommended in repeated genotyping. The EC method had a much lower false positive rate than did the GC method under different error rates. This study systematically expounded the impact of genotyping errors on linkage analysis, providing potential guidelines for improving the accuracy of linkage maps in the presence of genotyping errors. [ABSTRACT FROM AUTHOR]

Crop yield is a complex phenomenon, controlled by several genes, each contributing to the overall phenotypic expression; which is affected by environment and genotype x environment interactions. Identifying and mapping quantitative trait loci (QTLs), make tracing these traits during breeding possible and easy. The objective of this study was to identify QTLs associated with chickpea (Cicer arietinum L.) grain yield and its associated traits, under drought stress. The experiment was conducted using 188 F3:5 genotypes from ICCV 05107 x ICCV 94954 crosses. Genotypic data were from 49 polymorphic simple sequence repeat (SSR) markers; while phenotypic data were obtained from a field evaluation designed in a 19 x 10 alpha lattice. The study was replicated thrice on three sites, namely at Koibatek Agricultural Training Centre, Muserech; and at the Kenya Agricultural Research and Livestock Organisation, Marigat. Eight QTLs were mapped on a linkage map spanning a total length of 335.04 cM, with varying phenotypic variation expression (PVE%). These QTLs include, one each for days to maturity, 100-seed weight, and two each for above-ground biomass, harvest index, and grain yield. Five major QTLs having PVE ranging from 10.37 to 32.39%, were identified for days to maturity, 100-seed weight above-ground biomass, harvest index (HI), and grain yield. Four of the eight QTLs were mapped on linkage group 4 (LG4); days to maturity, 2 for above-ground biomass, and grain yield. The QTLs mapped are useful in genomic-assisted breeding for chickpea yield improvement. However, there is a need for marker saturation on LGs and specific genes identified for effective marker-assisted breeding. [ABSTRACT FROM AUTHOR]

Psidium guajava L. is an important fruit crop in the tropical and subtropical regions of the world. The advanced breeding methods are not employed for important commercial traits like peel and pulp color, seed hardiness, fruit size, etc., due to the scarcity of genome-wide molecular markers and high-density linkage maps. In this study, we employed single-nucleotide polymorphism (SNP) markers and identified quantitative trait loci (QTL) regions that are associated with color traits of leaf, peel, and pulp in the guava intervarietal mapping population. The mapping population was developed from the contrasting genotypes of fruit and leaf color. Variations in color among the segregating hybrids were recorded both visually and using a Color reader. A high-density linkage map of guava was constructed using the SNP markers from genotyping by sequencing (GBS) of 150 hybrid individuals of the cross 'Arka Poorna' (green) x 'Purple Local' (purple). The integrated linkage map consisted of 1426 SNPs mapped on 11 linkage groups (LG), spanning a total distance of around 730 cM with an average of 129.6 markers per LG. Through QTL analysis for color traits, a minor QTL region was identified for visually scored leaf color and peel color on LG1, whereas a major QTL was detected for pulp color in LG4. The Hunter color values (L* and, a*) also had major QTLs with overlapping marker intervals for leaf and peel colors, establishing the association of SNP markers to the trait. The QTLs harbored genes and transcription factors involved in lycopene and anthocyanin pigment biosynthesis. This is the first report of a high-density linkage map based on SNP markers in guava and QTL mapping for color characters in leaf, fruit peel and pulp. The genotyping information generated in this study can aid in genetic engineering and marker-assisted breeding in guava. [ABSTRACT FROM AUTHOR]

The reinvention of potato, from a tetraploid clonal crop into a diploid seed-based hybrid crop, requires insight in the mutational load, recombination landscape, and the genetic basis of fertility. Genomics-based breeding and QTL discovery rely on efficient genotyping strategies such as skim sequencing, to gather genotypic information. The application of skim sequencing to full-sib population of non-inbred parents remains challenging. Here, we report on an R implementation of the OutcrossSeq pipeline for diploids. We applied this pipeline to a large diploid skim sequenced potato population. We used the resulting bin-markers for the construction of high-density parent specific linkage maps, highlighting variation in parental recombination rate and structural variations. We subsequently explored transmission ratio distortion and non-independent assortment of alleles, indicative of large-effect deleterious mutations. Finally, we identified QTLs for seedling tuber yield in pots and pollen shed. This study showcases the range of genetic analyses, from marker inference, identification of transmission ratio distortion, and linkage map construction to QTL mapping, resulting in new insights that contribute to breeding diploid potato. [ABSTRACT FROM AUTHOR]

Abstract 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.

A large seed mutant, TG 89 having a 76.7% increment in hundred kernel weight in comparison to its parent TG 26, was isolated from an electron beam-induced mutagenized population. Studies based on environmental scanning electron microscopy of both parent and mutant revealed that the mutant seed cotyledon had significantly bigger cell size than parent. A mapping population with 122 F2 plants derived from the mutant and a distant normal seed genotype (ICGV 15007) was utilized to map the QTL associated with higher HKW. Bulk segregant analysis revealed putative association of three markers with this mutant large seed trait. Further, genotyping of F2 individuals with polymorphic markers detected 14 linkage groups with a map distance of 1053 cM. QTL analysis revealed a significant additive major QTL for the mutant large seed trait on linkage group A05 explaining 12.7% phenotypic variation for the seed size. This QTL was located between flanking markers AhTE333 and AhTE810 having a map interval of 4.7 cM which corresponds to 90.65 to 107.24 Mbp in A05 chromosome, respectively. Within this genomic fragment, an ortholog of the BIG SEEDS 1 gene was found at 102,476,137 bp. Real-time PCR revealed down-regulation of this BIG SEEDS 1 gene in the mutant indicating a loss of function mutation giving rise to a large seed phenotype. This QTL was validated in 11 advanced breeding lines having large seed size from this mutant but with varied genetic backgrounds. This validation showcased a highly promising selection accuracy of 90.9% for the marker-assisted selection. [ABSTRACT FROM AUTHOR]

Background: The Yellowfin tuna (Thunnus albacares) is a large tuna exploited by major fisheries in tropical and subtropical waters of all oceans except the Mediterranean Sea. Genomic studies of population structure, adaptive variation or of the genetic basis of phenotypic traits are needed to inform fisheries management but are currently limited by the lack of a reference genome for this species. Here we report a draft genome assembly and a linkage map for use in genomic studies of T. albacares. Methods and results: Illumina and PacBio SMRT sequencing were used in combination to generate a hybrid assembly that comprises 743,073,847 base pairs contained in 2,661 scaffolds. The assembly has a N50 of 351,587 and complete and partial BUSCO scores of 86.47% and 3.63%, respectively. Double-digest restriction associated DNA (ddRAD) was used to genotype the 2 parents and 164 of their F1 offspring resulting from a controlled breeding cross, retaining 19,469 biallelic single nucleotide polymorphism (SNP) loci. The SNP loci were used to construct a linkage map that features 24 linkage groups that represent the 24 chromosomes of yellowfin tuna. The male and female maps span 1,243.8 cM and 1,222.9 cM, respectively. The map was used to anchor the assembly in 24 super-scaffolds that contain 79% of the yellowfin tuna genome. Gene prediction identified 46,992 putative genes 20,203 of which could be annotated via gene ontology. Conclusions: The draft reference will be valuable to interpret studies of genome wide variation in T. albacares and other Scombroid species. [ABSTRACT FROM AUTHOR]

Shiitake mushroom (Lentinula edodes) is one of the most popular and widely cultivated mushrooms, especially in East Asia. Temperature is the critical factor affecting fruiting body induction. Due to climate change and increasing fuel costs, there is a growing demand for breeding L. edodes strains that fruit at higher temperatures. This study constructed a linkage map using 94 single spore isolates (SSIs) from the hybrid strain MCR 487 derived from crossing the high-temperature type (H-type) strain KRCF 1097 and the low to medium-temperature type (LM-type) strain KRCF 1101. Quantitative trait loci (QTLs) associated with higher-temperature fruiting ability were analyzed using 92 dikaryotic strains obtained from crossing SSIs from MCR 487 with the L-type strain KRCF 1098SS-2. Four QTLs (qTF1-4) controlling temperature for fruiting body induction were identified on the linkage map. Specific DNA marker was developed based on the sequence of the H-type qTF1, enabling the selection of strains capable of fruiting at higher temperatures. This study demonstrates that genome sequencing and QTL analysis can be used to obtain parental strains for breeding L. edodes strains suited for higher temperature cultivation, offering promising prospects for future breeding strategies. [ABSTRACT FROM AUTHOR]

The evolution of azole resistance in fungal pathogens presents a major challenge in both crop production and human health. Apple orchards across the world are faced with the emergence of azole fungicide resistance in the apple scab pathogen Venturia inaequalis. Target site point mutations observed in this fungus to date cannot fully explain the reduction in sensitivity to azole fungicides. Here, polygenic resistance to tebuconazole was studied across a population of V. inaequalis. Genotyping by sequencing allowed Quantitative Trait Loci (QTLs) mapping to identify the genetic components controlling this fungicide resistance. Dose-dependent genetic resistance was identified, with distinct genetic components contributing to fungicide resistance at different exposure levels. A QTL within linkage group seven explained 65% of the variation in the effective dose required to reduce growth by 50% (ED50). This locus was also involved in resistance at lower fungicide doses (ED10). A second QTL in linkage group one was associated with dose-dependent resistance, explaining 34% of variation at low fungicide doses (ED10), but did not contribute to resistance at higher doses (ED50 and ED90). Within QTL regions, non-synonymous mutations were observed in several ATP-Binding Cassette and Major Facilitator SuperFamily transporter genes. These findings provide insight into the mechanisms of fungicide resistance that have evolved in horticultural pathogens. Identification of resistance gene candidates supports the development of molecular diagnostics to inform management practices. [ABSTRACT FROM AUTHOR]

Italian ryegrass (Lolium multiflorum Lam.; IRG), an important Lolium species, is mainly cultivated in South Korea, Japan, China, and used as forage (pastes, hay, silage). Another important species of Lolium are perennial ryegrass (Lolium perenne L.; PRG), which is cultivated in Europe, United States of America, Australia, and New Zealand. The main character of the genus Lolium, which includes IRG and PRG, is its palatability; however, Lolium species exhibit summer depression during hot summers. The breeding goal of Festulolium, an interspecific and generic hybridization product of Lolium and Festuca, is to overcome the shortcomings of Lolium species. As such, pasture breeding is same varieties because polycrossing methods are used as five or more genetic resources, but there are some differences within the individual varieties. Amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers are used to investigate the geographical origins, ploidy levels, biological status, donor information, and flowering seasons of these varieties. Genetic markers are also used to construct linkage maps. Based on linkage map analyses, studies of Lolium quantitative trait loci (QTL) contributed to the breeding of varieties with improved tolerance or resistance to biological and non-biological stress factors. In this study, we summarize recent studies on the cultivation status of IRG in South Korea, linkage mapping of Lolium, and associated QTL analyses. We also discuss interspecific and generic hybridization of Lolium and Festuca. [ABSTRACT FROM AUTHOR]

Understanding the genetic basis of seed Ni and Mo is essential. Since soybean is a major crop in the world and a major source for nutrients, including Ni and Mo, the objective of the current research was to map genetic regions (quantitative trait loci, QTL) linked to Ni and Mo concentrations in soybean seed. A recombinant inbred line (RIL) population was derived from a cross between 'Forrest' and 'Williams 82' (F × W82). A total of 306 lines was used for genotyping using 5405 single nucleotides polymorphism (SNP) markers using Infinium SNP6K BeadChips. A two-year experiment was conducted and included the parents and the RIL population. One experiment was conducted in 2018 in North Carolina (NC), and the second experiment was conducted in Illinois in 2020 (IL). Logarithm of the odds (LOD) of ≥2.5 was set as a threshold to report identified QTL using the composite interval mapping (CIM) method. A wide range of Ni and Mo concentrations among RILs was observed. A total of four QTL (qNi-01, qNi-02, and qNi-03 on Chr 2, 8, and 9, respectively, in 2018, and qNi-01 on Chr 20 in 2020) was identified for seed Ni. All these QTL were significantly (LOD threshold > 2.5) associated with seed Ni, with LOD scores ranging between 2.71–3.44, and with phenotypic variance ranging from 4.48–6.97%. A total of three QTL for Mo (qMo-01, qMo-02, and qMo-03 on Chr 1, 3, 17, respectively) was identified in 2018, and four QTL (qMo-01, qMo-02, qMo-03, and qMo-04, on Chr 5, 11, 14, and 16, respectively) were identified in 2020. Some of the current QTL had high LOD and significantly contributed to the phenotypic variance for the trait. For example, in 2018, Mo QTL qMo-01 on Chr 1 had LOD of 7.8, explaining a phenotypic variance of 41.17%, and qMo-03 on Chr 17 had LOD of 5.33, with phenotypic variance explained of 41.49%. In addition, one Mo QTL (qMo-03 on Chr 14) had LOD of 9.77, explaining 51.57% of phenotypic variance related to the trait, and another Mo QTL (qMo-04 on Chr 16) had LOD of 7.62 and explained 49.95% of phenotypic variance. None of the QTL identified here were identified twice across locations/years. Based on a search of the available literature and of SoyBase, the four QTL for Ni, identified on Chr 2, 8, 9, and 20, and the five QTL associated with Mo, identified on Chr 1, 17, 11, 14, and 16, are novel and not previously reported. This research contributes new insights into the genetic mapping of Ni and Mo, and provides valuable QTL and molecular markers that can potentially assist in selecting Ni and Mo levels in soybean seeds. [ABSTRACT FROM AUTHOR]

Psidium guajava L. is an important fruit crop in the tropical and subtropical regions of the world. The advanced breeding methods are not employed for important commercial traits like peel and pulp color, seed hardiness, fruit size, etc., due to the scarcity of genome-wide molecular markers and high-density linkage maps. In this study, we employed single-nucleotide polymorphism (SNP) markers and identified quantitative trait loci (QTL) regions that are associated with color traits of leaf, peel, and pulp in the guava intervarietal mapping population. The mapping population was developed from the contrasting genotypes of fruit and leaf color. Variations in color among the segregating hybrids were recorded both visually and using a Color reader. A high-density linkage map of guava was constructed using the SNP markers from genotyping by sequencing (GBS) of 150 hybrid individuals of the cross ‘Arka Poorna’ (green) x ‘Purple Local’ (purple). The integrated linkage map consisted of 1426 SNPs mapped on 11 linkage groups (LG), spanning a total distance of around 730 cM with an average of 129.6 markers per LG. Through QTL analysis for color traits, a minor QTL region was identified for visually scored leaf color and peel color on LG1, whereas a major QTL was detected for pulp color in LG4. The Hunter color values (L* and, a*) also had major QTLs with overlapping marker intervals for leaf and peel colors, establishing the association of SNP markers to the trait. The QTLs harbored genes and transcription factors involved in lycopene and anthocyanin pigment biosynthesis. This is the first report of a high-density linkage map based on SNP markers in guava and QTL mapping for color characters in leaf, fruit peel and pulp. The genotyping information generated in this study can aid in genetic engineering and marker-assisted breeding in guava.

Horsegram (Macrotyloma uniflorum) - an important, self-pollinated food legume, however due to limited genomic and genetic resources the genetic improvement could not be achieved as compare to other major legumes. Our work aims at finding novel microsatellite markers and their use for the construction of a linkage map from 157 individuals of F9 recombinant inbred lines (RILs) of horsegram. The determinate growth habit and plant height are important traits for its suitability for different cropping systems. The genotypic data were generated by screening 2 395 molecular markers, of which 600 (25.05 %) polymorphic markers were selected. Two-hundred eighty-seven (287) markers were mapped on ten linkage groups (LGs) at a log of odds (LOD) of 3.5 straddling 796.76 cM with 2.78 cM of marker density. For the identification of the quantitative trait loci (QTLs), the phenotypic data recorded on the RILs for the plant height and growth habit were analysed using the statistical tools JoinMap®and Windows QTL cartographer, based on the composite interval mapping (CIM) technique. Across the ten linkage groups, we detected four QTLs (LOD ≥ 2.5) for four traits. All the traits were major QTLs as indicated by the percentage of phenotypic variance (PVE) (≥ 10%) that ranged from 13.5% to 40.3%, therefore, this is very important information which can be used in marker-assisted selection (MAS). The present genomic information generated in this orphan crop, thus, provides the base for genetic improvements by devising molecular breeding strategies.

Abstract Background For Asian seabass (Lates calcarifer, Bloch 1790) cultured at sea cages various aquatic pathogens, complex environmental and stress factors are considered as leading causes of disease, causing tens of millions of dollars of annual economic losses. Over the years, we conducted farm-based challenges by exposing Asian seabass juveniles to complex natural environmental conditions. In one of these challenges, we collected a total of 1,250 fish classified as either ‘sensitive’ or ‘robust’ individuals during the 28-day observation period. Results We constructed a high-resolution linkage map with 3,089 SNPs for Asian seabass using the double digest Restriction-site Associated DNA (ddRAD) technology and a performed a search for Quantitative Trait Loci (QTL) associated with robustness. The search detected a major genome-wide significant QTL for increased robustness in pathogen-infected marine environment on linkage group 11 (ASB_LG11; 88.9 cM to 93.6 cM) with phenotypic variation explained of 81.0%. The QTL was positioned within a > 800 kb genomic region located at the tip of chromosome ASB_LG11 with two Single Nucleotide Polymorphism markers, R1-38468 and R1-61252, located near to the two ends of the QTL. When the R1-61252 marker was validated experimentally in a different mass cross population, it showed a statistically significant association with increased robustness. The majority of thirty-six potential candidate genes located within the QTL have known functions related to innate immunity, stress response or disease. By utilizing this ddRAD-based map, we detected five mis-assemblies corresponding to four chromosomes, namely ASB_LG8, ASB_LG9, ASB_LG15 and ASB_LG20, in the current Asian seabass reference genome assembly. Conclusion According to our knowledge, the QTL associated with increased robustness is the first such finding from a tropical fish species. Depending on further validation in other stocks and populations, it might be potentially useful for selecting robust Asian seabass lines in selection programs.

Horsegram (Macrotyloma uniflorum) - an important, self-pollinated food legume, however due to limited genomic and genetic resources the genetic improvement could not be achieved as compare to other major legumes. Our work aims at finding novel microsatellite markers and their use for the construction of a linkage map from 157 individuals of F9 recombinant inbred lines (RILs) of horsegram. The determinate growth habit and plant height are important traits for its suitability for different cropping systems. The genotypic data were generated by screening 2 395 molecular markers, of which 600 (25.05 %) polymorphic markers were selected. Two-hundred eighty-seven markers were mapped on ten linkage groups (LGs) at a log of odds (LOD) of 3.5 straddling 796.76 cM with 2.78 cM of marker density. For the identification of the quantitative trait loci (QTLs), the phenotypic data recorded on the RILs for the plant height and growth habit were analysed using the statistical tools JoinMap® and Windows QTL cartographer, based on the composite interval mapping (CIM) technique. Across the ten linkage groups, we detected four QTLs (LOD ≥ 2.5) for four traits. All the traits were major QTLs as indicated by the percentage of phenotypic variance (PVE) (≥ 10%) that ranged from 13.5% to 40.3%, therefore, this is very important information which can be used in marker-assisted selection (MAS). The present genomic information generated in this orphan crop, thus, provides the base for genetic improvements by devising molecular breeding strategies. [ABSTRACT FROM AUTHOR]

Soybean seed sugars are among the most abundant beneficial compounds for human and animal consumption in soybean seeds. Higher seed sugars such as sucrose are desirable as they contribute to taste and flavor in soy-based food. Therefore, the objectives of this study were to use the 'Forrest' by 'Williams 82' (F × W82) recombinant inbred line (RIL) soybean population (n = 309) to identify quantitative trait loci (QTLs) and candidate genes that control seed sugar (sucrose, stachyose, and raffinose) contents in two environments (North Carolina and Illinois) over two years (2018 and 2020). A total of 26 QTLs that control seed sugar contents were identified and mapped on 16 soybean chromosomes (chrs.). Interestingly, five QTL regions were identified in both locations, Illinois and North Carolina, in this study on chrs. 2, 5, 13, 17, and 20. Amongst 57 candidate genes identified in this study, 16 were located within 10 Megabase (MB) of the identified QTLs. Amongst them, a cluster of four genes involved in the sugars' pathway was collocated within 6 MB of two QTLs that were detected in this study on chr. 17. Further functional validation of the identified genes could be beneficial in breeding programs to produce soybean lines with high beneficial sucrose and low raffinose family oligosaccharides. [ABSTRACT FROM AUTHOR]

Obligate parthenogenesis (OP) is often thought to evolve by disruption of reductional meiosis and suppression of crossover recombination. In the crustacean Daphnia pulex , OP lineages, which have evolved from cyclical parthenogenetic (CP) ancestors, occasionally produce males that are capable of reductional meiosis. Here, by constructing high-density linkage maps, we find that these males show only slightly and nonsignificantly reduced recombination rates compared to CP males and females. Both meiosis disruption and recombination suppression are therefore sex-limited (or partly so), which speaks against the evolution of OP by disruption of a gene that is essential for meiosis or recombination in both sexes. The findings may be explained by female-limited action of genes that suppress recombination, but previously identified candidate genes are known to be expressed in both sexes. Alternatively, and equally consistent with the data, OP might have evolved through a reuse of the parthenogenesis pathways already present in CP and through their extension to all events of oogenesis. The causal mutations for the CP to OP transition may therefore include mutations in genes involved in oogenesis regulation and may not necessarily be restricted to genes of the "meiosis toolkit." More generally, our study emphasizes that there are many ways to achieve asexuality, and elucidating the possible mechanisms is key to ultimately identify the genes and traits involved. [ABSTRACT FROM AUTHOR]

Background: For Asian seabass (Lates calcarifer, Bloch 1790) cultured at sea cages various aquatic pathogens, complex environmental and stress factors are considered as leading causes of disease, causing tens of millions of dollars of annual economic losses. Over the years, we conducted farm-based challenges by exposing Asian seabass juveniles to complex natural environmental conditions. In one of these challenges, we collected a total of 1,250 fish classified as either 'sensitive' or 'robust' individuals during the 28-day observation period. Results: We constructed a high-resolution linkage map with 3,089 SNPs for Asian seabass using the double digest Restriction-site Associated DNA (ddRAD) technology and a performed a search for Quantitative Trait Loci (QTL) associated with robustness. The search detected a major genome-wide significant QTL for increased robustness in pathogen-infected marine environment on linkage group 11 (ASB_LG11; 88.9 cM to 93.6 cM) with phenotypic variation explained of 81.0%. The QTL was positioned within a > 800 kb genomic region located at the tip of chromosome ASB_LG11 with two Single Nucleotide Polymorphism markers, R1-38468 and R1-61252, located near to the two ends of the QTL. When the R1-61252 marker was validated experimentally in a different mass cross population, it showed a statistically significant association with increased robustness. The majority of thirty-six potential candidate genes located within the QTL have known functions related to innate immunity, stress response or disease. By utilizing this ddRAD-based map, we detected five mis-assemblies corresponding to four chromosomes, namely ASB_LG8, ASB_LG9, ASB_LG15 and ASB_LG20, in the current Asian seabass reference genome assembly. Conclusion: According to our knowledge, the QTL associated with increased robustness is the first such finding from a tropical fish species. Depending on further validation in other stocks and populations, it might be potentially useful for selecting robust Asian seabass lines in selection programs. [ABSTRACT FROM AUTHOR]

Cunninghamia lanceolata is one of the most important tree species in China due to its significance both in economy and ecology. The aims of the present study were to construct a high-density genetic map and identify a quantitative trait locus (QTL) for C. lanceolata. In this study, an F1 population comprising 81 individuals was developed. Using specific length amplified fragment sequencing (SLAF-seq) technology, a total of 254,899 loci were found to be polymorphic. After linkage analysis, 2574 markers were used to construct genetic linkage maps. Specifically, 1632 markers were allocated to 11 linkage groups (LGs) for the female map, 1038 for the male map, and 2574 for the integrated map. The integrated map consisted of 4596 single-nucleotide polymorphisms (SNPs) loci, resulting in an average of 1.79 SNP loci per SLAF marker. The marker coverage was 1665.76 cM for the female map, 1436.39 cM for the male map, and 1748.40 cM for the integrated map. The average interval between two adjacent mapped markers was 1.03 cM, 1.40 cM, and 0.68 cM for the female map, male map, and integrated map, respectively. Using the integrated map, we performed interval mapping (logarithm of odds, LOD > 2.0) to detect traits of interest. We identified a total of 2, 1, 2, 5, 1, 2, 1, and 3 QTLs for diameter at breast height, heartwood diameter, heartwood proportion, heartwood a*, heartwood b*, heartwood L*, sapwood a*, and sapwood L*, respectively. The number of markers associated with each QTL ranged from 1 to 14, and each marker explained phenotypic variances ranging from 12.70% to 23.60%. Furthermore, a common QTL was identified for diameter at breast height and heartwood color a*, while another common QTL was observed for heartwood color L* and heartwood color a*. These findings suggest possible pleiotropic effects of the same genes on these traits. In conclusion, we successfully constructed high-density genetic maps for C. lanceolata using the SLAF-seq method with an F1 population. Notably, these linkage maps represent the most comprehensive and densest ones available to date for C. lanceolata and will facilitate future chromosome assignments for C. lanceolata whole-genome sequencing. These identified QTLs will serve as a valuable resource for conducting fine-scale QTL mapping and implementing marker-assisted selection in C. lanceolata, particularly for growth and wood-color traits. [ABSTRACT FROM AUTHOR]

Melon (Cucumis melo L.) is an important horticultural cash crop and its quality traits directly affect consumer choice and market price. These traits are controlled by genetic as well as environmental factors. In this study, a quantitative trait locus (QTL) mapping strategy was used to identify the potential genetic loci controlling quality traits of melons (i.e., exocarp and pericarp firmness and soluble solid content) based on newly derived whole-genome single nucleotide polymorphism-based cleaved amplified polymorphic sequence (SNP-CAPS) markers. Specifically, SNPs of two melon varieties, M4-5 and M1-15, as revealed by whole-genome sequencing, were converted to the CAPS markers, which were used to construct a genetic linkage map comprising 12 chromosomes with a total length of 1414.88 cM, in the F2 population of M4-5 and M1-15. The six identified QTLs included: SSC6.1 and SSC11.1 related to soluble solid content; EF12.1 associated with exocarp firmness; and EPF3.1, EPF3.2 and EPF7.1 related to edible pericarp firmness. These genes were located on five chromosomes (3, 6, 7, 11, and 12) in the flanking regions of the CAPS markers. Moreover, the newly developed CAPS markers will be useful in guiding genetic engineering and molecular breeding in melon.

IntroductionRice is an important crop that is considered a staple meal for 2.7 billion people worldwide. Therefore, the demand for it will increase with the increase of population. Environmental constraints always pose a serious threat to crop production, including rice. Drought is one of the most important challenges that limits the production of high-yielding cultivars in arid and rainfed areas. Global warming has also become a factor in limiting rice production in rain-dependent areas. Therefore, researchers are looking for a way to stabilize rice production in arid regions. In this study, informative markers related to the desired agronomic traits were identified in 59 rice genotypes using microsatellite marking system.Materials and methodsIn order to evaluate the tolerance of rice genotypes to drought stress and to identify tolerant and sensitive genotypes, 59 genotypes received from the National Rice Research Institute and the International Rice Research Institute in a randomized complete block design with three replications in two separate conditions, without Stress (flood) and drought stress were performed in a research farm located in Aliabad Katoul city in 2013. In both conditions (normal and drought stress), the genotypes were planted in five rows of 25 × 25 cm in rows one meter long. Thirty days after planting in the nursery, healthy and strong seedlings were transferred to the main land. The required agronomic operations were carried out equally during the growth and development period of the plants under stress and normal conditions and only in terms of irrigation of the experimental field in both flood and stress environments, until the tillering stage of the cultivars were equally flooded. Then, to create stress, irrigation was done from 40 days after transplanting (maximum tillering stage) to the end of the growing season at 25-day intervals. Phenotypic values of grain yield and 1000-grain weight were measured under two conditions according to standard guidelines for evaluation of traits in rice. In order to investigate the relationship between agronomic traits and microsatellite markers with 59 rice genotypes out of 36 microsatellite molecular markers were performed in the Plant Breeding and Genetics Laboratory of Gonbad Kavous University, Faculty of Agriculture and Natural Resources. Young leaves of 21-day-old seedlings were extracted in four-leaf stage using CTAB method. Touchdown PCR reaction was studied and evaluated randomly using 36 microsatellite primers for 3 markers from each chromosome. To separate PCR products, 6% polyacrylamide gel electrophoresis was used and to reveal the banding pattern, silver nitrate staining method was used. The content of polymorphic information was calculated. The relationship between molecular data and traits of studied rice genotypes was investigated using multiple regression. Thus, each quantitative trait was considered as a dependent variable and microsatellite markers were considered as independent variables.Results and discussionThe average content of polymorphic corrections (PIC) was estimated to be 0.58, which showed RM 5647 with 0.81 the highest and RM 6022 with 0.32 the lowest polymorphism (PIC). The results of stepwise regression analysis showed that a total of 90 markers for normal conditions and 69 markers for drought stress conditions for morphological traits were identified. Under normal conditions, the number of spikes and the number of days to flowering with 9 markers and under drought stress, the weight of the cluster with 9 markers showed the most positive markers. The most explanation for variation in normal conditions is related to the total number of grains (0.83) by gene loci RM6324-E, RM5652-E, RM5761-D, RM6179-F, RM549-B, RM462-B, RM7420-D Explained. In drought stress conditions, the most explanation for variation related to panicle weight (0.70) by gene loci RM519-D, RM7545-A, RM6179-E, RM7118-G, RM3525-B, RM5761-B, RM38-C, RM7091-A, RM5647-B explained.ConclusionThe results showed that some markers are associated with more than one trait, which indicates that these traits are very closely related to each other or may be influenced by multi-effect genes. To understand this, it is necessary to develop transgressive generations and linkage.

Abstract Background Sex determination occurs across animal species, but most of our knowledge about its mechanisms comes from only a handful of bilaterian taxa. This limits our ability to infer the evolutionary history of sex determination within animals. Results In this study, we generated a linkage map of the genome of the colonial cnidarian Hydractinia symbiolongicarpus and used it to demonstrate that this species has an XX/XY sex determination system. We demonstrate that the X and Y chromosomes have pseudoautosomal and non-recombining regions. We then use the linkage map and a method based on the depth of sequencing coverage to identify genes encoded in the non-recombining region and show that many of them have male gonad-specific expression. In addition, we demonstrate that recombination rates are enhanced in the female genome and that the haploid chromosome number in Hydractinia is n = 15. Conclusions These findings establish Hydractinia as a tractable non-bilaterian model system for the study of sex determination and the evolution of sex chromosomes.

BACKGROUND:Unravelling the genetic architecture of agronomic traits in walnut such as budbreak date and bearing habit, is crucial for climate change adaptation and yield improvement. A Genome-Wide Association Study (GWAS) using multi-locus models was conducted in a panel of 170 walnut accessions genotyped using the Axiom™ J. regia 700 K SNP array, with phenological data from 2018, 2019 and legacy data. These accessions come from the INRAE walnut germplasm collection which is the result of important prospecting work performed in many countries around the world. In parallel, an F1 progeny of 78 individuals segregating for phenology-related traits, was genotyped with the same array and phenotyped for the same traits, to construct linkage maps and perform Quantitative Trait Loci (QTLs) detection. RESULTS:Using GWAS, we found strong associations of SNPs located at the beginning of chromosome 1 with both budbreak and female flowering dates. These findings were supported by QTLs detected in the same genomic region. Highly significant associated SNPs were also detected using GWAS for heterodichogamy and lateral bearing habit, both on chromosome 11. We developed a Kompetitive Allele Specific PCR (KASP) marker for budbreak date in walnut, and validated it using plant material from the Walnut Improvement Program of the University of California, Davis, demonstrating its effectiveness for marker-assisted selection in Persian walnut. We found several candidate genes involved in flowering events in walnut, including a gene related to heterodichogamy encoding a sugar catabolism enzyme and a cell division related gene linked to female flowering date. CONCLUSIONS:This study enhances knowledge of the genetic architecture of important agronomic traits related to male and female flowering processes and lateral bearing in walnut. The new marker available for budbreak date, one of the most important traits for good fruiting, will facilitate the selection and development of new walnut cultivars suitable for specific climates.

Simple Summary: Although lentil is not as popular as other legumes, it is a climate-resilient legume crop because of its high protein content, nitrogen fixation, and abiotic stress tolerance ability. Even though it can be grown on almost every continent and is distributed globally, the use of existing genetic diversity in marker-assisted selection is still limited. In this study, novel SSR markers needed in lentil were identified using a next-generation sequencing approach. In this study, we created a ready-to-use SSR library for genetic diversity studies and breeding and evaluated the effectiveness of the obtained SSR markers in a recombinant inbred line (RIL) population. Simple sequence repeats (SSRs) are highly versatile markers in genetic diversity analysis and plant breeding, making them widely applicable. They hold potential in lentil (Lens culinaris) breeding for genetic diversity analysis, marker-assisted selection (MAS), and linkage mapping. However, the availability and diversity of SSR markers in lentil is limited. We used next-generation sequencing (NGS) technology to develop SSR markers in lentil. NGS allowed us to identify regions of the lentil genome that contained SSRs. Illumina Hiseq-2000 sequencing of the lentil genotype "Karacadağ" resulted in 1,727,734 sequence reads comprising more than 48,390 Mb, and contigs were mined for SSRs, resulting in the identification of a total of 8697 SSR motifs. Among these, dinucleotide repeats were the most abundant (53.38%), followed by trinucleotides (30.38%), hexanucleotides (6.96%), tetranucleotides (6.59%), and pentanucleotides (3.19%). The most frequent repeat in dinucleotides was the TC (21.80%), followed by the GA (17.60%). A total of 2000 primer pairs were designed from these motifs, and 458 SSR markers were validated following their amplified PCR products. A linkage map was constructed using these new SSRs with high linkage disequilibrium (209) and previously known SSRs (11). The highest number of SSR markers (43) was obtained in LG2, while the lowest number of SSR markers (19) was obtained in LG7. The longest linkage group (LG) was LG2 (86.84 cM), whereas the shortest linkage group was LG7 (53.46 cM). The average length between markers ranged from 1.86 cM in LG1 to 2.81 cM in LG7, and the map density was 2.16 cM. The developed SSRs and created linkage map may provide useful information and offer a new library for genetic diversity analyses, linkage mapping studies, and lentil breeding programs. [ABSTRACT FROM AUTHOR]

The production of Brassica oleracea, an important vegetable crop, is severely affected by black rot disease caused by the bacterial pathogen Xanthomonas campestris pv. campestris. Resistance to race 1, the most virulent and widespread race in B. oleracea, is under quantitative control; therefore, identifying the genes and genetic markers associated with resistance is crucial for developing resistant cultivars. Quantitative trait locus (QTL) analysis of resistance in the F2 population developed by crossing the resistant parent BR155 with the susceptible parent SC31 was performed. Sequence GBS approach was used to develop a genetic linkage map. The map contained 7,940 single nucleotide polymorphism markers consisting of nine linkage groups spanning 675.64 cM with an average marker distance of 0.66 cM. The F2:3 population (N = 126) was evaluated for resistance to black rot disease in summer (2020), fall (2020), and spring (2021). QTL analysis, using a genetic map and phenotyping data, identified seven QTLs with LOD values between 2.10 and 4.27. The major QTL, qCaBR1, was an area of overlap between the two QTLs identified in the 2nd and 3rd trials located at C06. Among the genes located in the major QTL interval, 96 genes had annotation results, and eight were found to respond to biotic stimuli. We compared the expression patterns of eight candidate genes in susceptible (SC31) and resistant (BR155) lines using qRT-PCR and observed their early and transient increases or suppression in response to Xanthomonas campestris pv. campestris inoculation. These results support the involvement of the eight candidate genes in black rot resistance. The findings of this study will contribute towards marker-assisted selection, additionally the functional analysis of candidate genes may elucidate the molecular mechanisms underlying black rot resistance in B. oleracea. [ABSTRACT FROM AUTHOR]

Downy mildew (DM) is a major foliar disease globally causing great economic loss in melon production. Utilizing disease-resistant cultivars is the most efficient approach for disease control, while discovery of disease-resistant genes is crucial for the success of DM-resistant breeding. To address this problem, two F2 populations were constructed using the DM-resistant accession PI 442177 in this study, and QTLs conferring DM resistance were mapped using linkage map and QTL-seq analysis, respectively. A high-density genetic map with the length of 1096.7 cM and density of 0.7 cM was generated by using the genotyping-bysequencing data of a F2 population. A major QTL DM9.1 with the phenotypic variance explained proportion of 24.3-37.7% was consistently detected at the early, middle, and late growth stages using the genetic map. QTL-seq analyses on the two F2 populations also validated the presence of DM9.1. Kompetitive Allele-Specific PCR (KASP) assay was further carried out to fine map DM9.1 into 1.0 Mb interval. A KASP marker co-segregating with DM9.1 was successfully developed. These results not only provided valuable information for DMresistant gene cloning, but also offered useful markers for melon DM-resistant breeding programs. [ABSTRACT FROM AUTHOR]

A mapping population, recombinant inbred lines (RILs), was developed from the cross of aphid susceptible cv. PL426 and resistant genotype BK 9816 to identify genetic loci conferring corn leaf aphid (CLA) Rhopalosiphum maidis (Fitch) resistance in barley. RILs showed wide range of aphid infestation (2.2–65.0 aphids/tiller) indicating quantitative inheritance of aphid resistance. All the traits related to aphid resistance, viz. aphid fecundity, chlorophyll content and tiller dry weight of plants, also showed quantitative variation. RILs were further genotyped by sequencing to identify SNPs covering whole barley genome. A total of 605 high-quality bin-mapped SNPs were positioned in seven linkage groups with a total map length of 1208.9 cM and average marker distance of 2.02 cM. Composite interval mapping identified five different quantitative trait loci (QTLs) on chromosomes 1H, 3H, 5H and 6H associated with aphid resistance and related traits. Two QTLs on chromosome 6H, QCLA.pau-6H.1 and QCLA.pau-6H.2, were mapped at genetic intervals of 1.7 cM and 4.9 cM, respectively. QCLA.pau-6H.1 explained 9.76 percent of phenotypic variation for aphid count and 12.78 percent for aphid score. Similarly, QCLA.pau-6H.2 explained 10.96 percent of phenotypic variation for aphid score. The QTL for chlorophyll content and tiller dry weight spanned the same region of QCLA.pau-6H.2, explained 13.08 and 13.20 percent phenotypic variation, respectively. Another QTL associated with chlorophyll content was mapped on chromosome 3H at 3.6-cM interval explained 12.90 percent phenotypic variation. In addition, QTL regions were also identified for aphid resistance and related traits on chromosome 1H and 5H. [ABSTRACT FROM AUTHOR]

The maize weevil, Sitophilus zeamais, is a worldwide pest that disproportionately affects subsistence farmers in developing countries. Damage from this pest threatens food security in these communities as widely available and effective control methods are lacking. With advances over the last decade in the development of genetic pest management techniques, addressing pest issues at the ecosystem level as opposed to the farm level may be a possibility. However, pest species selected for genetic management techniques require a well-characterized genome and few genomic tools have been developed for S. zeamais. Here, we have measured the genome size and developed the first genetic linkage map for this species. The genome size was determined using flow cytometry as 682 Mb and 674 Mb for females and males, respectively. The linkage map contains 11 linkage groups, which correspond to the 10 autosomes and 1 X-chromosome found in the species and it contains 1,121 SNPs. This linkage map will be useful for assembling a complete genome for S. zeamais. [ABSTRACT FROM AUTHOR]

Chili pepper (Capsicum annuum L.) is one of the oldest and most phenotypically diverse pre-Columbian crops of the Americas. Despite the abundance of genetic resources, the use of wild germplasm and landraces in chili pepper breeding is limited. A better understanding of the evolutionary history in chili peppers, particularly in the context of traits of agronomic interest, can contribute to future improvement and conservation of genetic resources. In this study, an F2:3 mapping population derived from a cross between a C. annuum wild accession (Chiltepin) and a cultivated variety (Puya) was used to identify genomic regions associated with 19 domestication and agronomic traits. A genetic map was constructed consisting of 1023 single nucleotide polymorphism (SNP) markers clustered into 12 linkage groups and spanning a total of 1,263.87 cM. A reciprocal translocation that differentiates the domesticated genome from its wild ancestor and other related species was identified between chromosomes 1 and 8. Quantitative trait locus (QTL) analysis detected 20 marker-trait associations for 13 phenotypes, from which 14 corresponded to previously identified loci, and six were novel genomic regions related to previously unexplored domestication-syndrome traits, including form of unripe fruit, seedlessness, deciduous fruit, and growth habit. Our results revealed that the genetic architecture of Capsicum domestication is similar to other domesticated species with few loci with large effects, the presence of QTLs clusters in different genomic regions, and the predominance of domesticated recessive alleles. Our analysis indicates the domestication process in chili pepper has also had an effect on traits not directly related to the domestication syndrome. The information obtained in this study provides a more complete understanding of the genetic basis of Capsicum domestication that can potentially guide strategies for the exploitation of wild alleles. [ABSTRACT FROM AUTHOR]

BACKGROUND:Recent research has revealed that polymorphic behavioral strategies shape intra-and interspecific interactions and contribute to fitness in many animal species. A better understanding of the proximate mechanisms underlying these behavioral syndromes will enhance our grasp this phenomenon. Spiders in the genus Anelosimus exhibit inter-individual behavioral variation on several axes: individuals have consistent responses to stimuli (e.g. bold vs. shy individuals) and they are subsocial (exhibiting extended maternal care and sibling cooperation) across most of their range, but they sometimes form permanent social groups in northern temperate regions. Here, we seek genetic variants associated with boldness and with social structure in a socially polymorphic population of the spider Anelosimus studiosus. We also develop preliminary genomic resources, including a genome assembly and linkage map, that support this and future genomic research on this group. RESULTS:Remarkably, we identify a small genomic scaffold (~ 1200 bp) that harbors seven single nucleotide polymorphisms (SNPs) associated with boldness. Moreover, heterozygotes are less common than expected based on Hardy-Weinberg equilibrium, suggesting that either assortative mating or selection against heterozygotes may be occurring in this system. We find no loci significantly associated with social organization. Our draft genome assembly allows us to localize SNPs of interest in this study and to carry out genetic comparisons with other published genomes, although it remains highly fragmented. CONCLUSIONS:By identifying a locus associated with a well-studied animal personality trait, this study opens up avenues for future research to link behavioral studies of animal personality with genotype and fitness.

The silkworm (Bombyx mori) is an important lepidopteran model insect and an industrial domestic animal traditionally used for silk production. Here, we report the genome assembly of an improved Japanese strain Nichi01, in which the cocoon yield is comparable to that of commercial silkworm strains. The integration of PacBio Sequel II long-read and ddRAD-seq-based high-density genetic linkage map achieved the highest quality genome assembly of silkworms to date; 22 of the 28 pseudomolecules contained telomeric repeats at both ends, and only four gaps were present in the assembly. A total of 452 Mbp of the assembly with an N50 of 16.614 Mbp covered 99.3% of the complete orthologs of the lepidopteran core genes. Although the genome sequence of Nichi01 and that of the previously reported low-yielding tropical strain p50T assured their accuracy in most regions, we corrected several regions, misassembled in p50T, in our assembly. A total of 18,397 proteins were predicted using over 95 Gb of mRNA-seq derived from 10 different organs, covering 96.9% of the complete orthologs of the lepidopteran core genes. The final assembly and annotation files are available in KAIKObase (https://kaikobase.dna.affrc.go.jp/index.html) along with a genome browser and BLAST searching service, which would facilitate further studies and the breeding of silkworms and other insects. [ABSTRACT FROM AUTHOR]

The unique color and type characteristics of watermelon fruits are regulated by many molecular mechanisms. However, it still needs to be combined with more abundant genetic data to fine-tune the positioning. We assembled genomes of two Korean inbred watermelon lines (cv. 242-1 and 159-1) with unique color and fruit-type characteristics and identified 23,921 and 24,451 protein-coding genes in the two genomes, respectively. To obtain more precise results for further study, we resequenced one individual of each parental line and an F2 population composed of 87 individuals. This identified 1,539 single-nucleotide polymorphisms (SNPs) and 80 InDel markers that provided a high-density genetic linkage map with a total length of 3,036.9 cM. Quantitative trait locus mapping identified 15 QTLs for watermelon fruit quality-related traits, including b-carotene and lycopene content in fruit flesh, fruit shape index, skin thickness, flesh color, and rind color. By investigating the mapping intervals, we identified 33 candidate genes containing variants in the coding sequence. Among them, Cla97C01G008760 was annotated as a phytoene synthase with a singlenucleotide variant (A!G) in the first exon at 9,539,129 bp of chromosome 1 that resulted in the conversion of a lysine to glutamic acid, indicating that this gene might regulate flesh color changes at the protein level. These findings not only prove the importance of a phytoene synthase gene in pigmentation but also explain an important reason for the color change of watermelon flesh. [ABSTRACT FROM AUTHOR]

Karyotypes are generally conserved between closely related species and large chromosome rearrangements typically have negative fitness consequences in heterozygotes, potentially driving speciation. In the order Lepidoptera, most investigated species have the ancestral karyotype and gene synteny is often conserved across deep divergence, although examples of extensive genome reshuffling have recently been demonstrated. The genus Leptidea has an unusual level of chromosome variation and rearranged sex chromosomes, but the extent of restructuring across the rest of the genome is so far unknown. To explore the genomes of the wood white (Leptidea) species complex, we generated eight genome assemblies using a combination of 10X linked reads and HiC data, and improved them using linkage maps for two populations of the common wood white (L. sinapis) with distinct karyotypes. Synteny analysis revealed an extensive amount of rearrangements, both compared to the ancestral karyotype and between the Leptidea species, where only one of the three Z chromosomes was conserved across all comparisons. Most restructuring was explained by fissions and fusions, while translocations appear relatively rare. We further detected several examples of segregating rearrangement polymorphisms supporting a highly dynamic genome evolution in this clade. Fusion breakpoints were enriched for LINEs and LTR elements, which suggests that ectopic recombination might be an important driver in the formation of new chromosomes. Our results show that chromosome count alone may conceal the extent of genome restructuring and we propose that the amount of genome evolution in Lepidoptera might still be underestimated due to lack of taxonomic sampling. [ABSTRACT FROM AUTHOR]

Tobacco (Nicotiana tabacum L.) is an economic crop and a model organism for studies on plant biology and genetics. A population of 271 recombinant inbred lines (RIL) derived from K326 and Y3, two elite flue-cured tobacco parents, has been constructed to investigate the genetic basis of agronomic traits in tobacco. Six agronomic traits including natural plant height (nPH), natural leaf number (nLN), stem girth (SG), inter-node length (IL), length of the largest leaf (LL) and width of the largest leaf (LW) were measured in seven environments, spanning the period between 2018 and 2021. We firstly developed an integrated SNP-indel-SSR linkage map with 43,301 SNPs, 2,086 indels and 937 SSRs, which contained 7,107 bin markers mapped on 24 LGs and covered 3334.88 cM with an average genetic distance of 0.469cM. Based on this high-density genetic map, a total of 70 novel QTLs were detected for six agronomic traits by a full QTL model using the software QTLNetwork, of which 32 QTLs showed significant additive effects, 18 QTLs showed significant additive-by-environment interaction effects, 17 pairs showed significant additive-by-additive epistatic effects and 13 pairs showed significant epistasis-by-environment interaction effects. In addition to additive effect as a major contributor to genetic variation, both epistasis effects and genotype-byenvironment interaction effects played an important role in explaining phenotypic variation for each trait. In particular, qnLN6-1 was detected with considerably large main effect and high heritability (h2a =34.80%). Finally, four genes including Nt16g00284.1, Nt16g00767.1, Nt16g00853.1, Nt16g00877.1 were predicted as pleiotropic candidate genes for five traits. [ABSTRACT FROM AUTHOR]

Background: Sex determination occurs across animal species, but most of our knowledge about its mechanisms comes from only a handful of bilaterian taxa. This limits our ability to infer the evolutionary history of sex determination within animals. Results: In this study, we generated a linkage map of the genome of the colonial cnidarian Hydractinia symbiolongicarpus and used it to demonstrate that this species has an XX/XY sex determination system. We demonstrate that the X and Y chromosomes have pseudoautosomal and non-recombining regions. We then use the linkage map and a method based on the depth of sequencing coverage to identify genes encoded in the non-recombining region and show that many of them have male gonad-specific expression. In addition, we demonstrate that recombination rates are enhanced in the female genome and that the haploid chromosome number in Hydractinia is n = 15. Conclusions: These findings establish Hydractinia as a tractable non-bilaterian model system for the study of sex determination and the evolution of sex chromosomes. [ABSTRACT FROM AUTHOR]

Black spot disease (PBS) caused by Alternaria alternata is an economic disease of pear (Pyrus pyrifolia Nakai). Developing cultivars with durable PBS resistance traits is an important research objective for improving pear germplasm. The Deshengxiang is a popular pear variety in China and resistant to PBS. This study aimed to detect quantitative trait loci (QTL) associated with PBS resistance trait in pear and determine closely linked molecular markers by specific locus amplified fragment sequencing (SLAF-seq). F1 population resulting from a cross between "Deshengxiang" (female) and "Guiguan," a susceptible (male) variety, was developed and evaluated in 2016 and 2017. SLAF technology was used to discover SNPs in the F1 individuals and subsequently a high-density genetic linkage map for PBS resistance was constructed which contained 17,604 SNP markers. Based on the linkage map, the markers were distributed into 17 linkage groups, spanning 1548.48 cM, with a mean marker distance of 0.09 cM, representing the densest genetic map of the genus Pyrus. QTL analysis of PBS resistance identified a locus strongly related to PBS resistance at 77.68 ~ 112.99 cM on linkage group 15, which was further narrowed down to 93.79 ~ 112.99 cM. Two markers, Marker94293 and Marker94206, located at 97.47 and 102.93 cM, were closely associated with PBS resistance, with a Δ (SNP index) value of 0.46. Co-localization of QTL interval, bioinformatics analysis, and functional annotation revealed PBS putative candidate genes. Overall, the high-density pear linkage map is a suitable reference for mapping PBS resistance trait, QTL, and genes identified in this study contribute information that could be useful for PBS improvement in pear. [ABSTRACT FROM AUTHOR]

Main conclusion: The present review illustrates a comprehensive overview of the start codon targeted (SCoT) polymorphism marker and their utilization in various applications related to genetic and genomic studies. Start codon targeted (SCoT) polymorphism marker, a targeted fingerprinting marker technique, has gained considerable importance in plant genetics, genomics, and molecular breeding due to its many desirable features. SCoT marker targets the region flanking the start codon, a highly conserved region in plant genes. Therefore, it can distinguish genetic variations in a specific gene that link to a specific trait. It is a simple, novel, cost-effective, highly polymorphic, and reproducible molecular marker for which there is no need for prior sequence information. In the recent past, SCoT markers have been employed in many commercially important and underutilized plant species for a variety of applications, including genetic diversity analysis, interspecific/generic genetic relationships, cultivar/hybrid/species identification, sex determination, construction of linkage map, association mapping/analysis, differential gene expression, and genetic fidelity analysis of tissue culture-raised plants. The main aim of this review is to provide up-to-date information on SCoT markers and their application in many commercially important and underutilized plant species, mainly progress made in the last 8–10 years. [ABSTRACT FROM AUTHOR]