Validation Of Quantitative Trait Loci Research Articles

Validation of quantitative trait loci for biofortification traits and variability research on agro-morphological, physiological, and quality traits in dicoccum wheat (Triticum dicoccum Schrank.)

Validation of quantitative trait loci for biofortification traits and variability research on agro-morphological, physiological, and quality traits in dicoccum wheat (Triticum dicoccum Schrank.)

Identification and validation of novel QTL associated with powdery mildew (Golovinomyces cichoracearum (DC.) V.P. Heluta.) resistance in sunflower (Helianthus annuus L.)

Sunflower (Helianthus annuus L.; 2n=2x=34) is an important annual oilseed crop cultivated worldwide. In the recent past, powdery mildew disease has evolved as a significant constraint for yield and quality production in sunflower crop. Hence, a F2 mapping population was developed from a cross COSF 15B x IR6 to identify quantitative trait loci (QTL) associated with powdery mildew resistance in sunflower. F2:4 recombinant lines of this population were screened along with parents and scored for powdery mildew resistance during Dec-Feb 2021-22. Per cent disease index (PDI) was calculated from the data taken 75 days after sowing (DAS) using a 0-9 score scale. Genetic linkage map was constructed with 92 SSR markers which covered 1053.3 cM. The QTL analysis resulted in the detection of one QTL in LG10 for PDI. It has 2.54 LOD and accounted for 12.25 per cent phenotypic variation. The flanking markers viz., ORS684 and ORS1110 have a flanking distance of 1.7 cM and validated with a set ofpopulation revealed that 24% and 28% PVE respectively. Hence, it may be concluded that markers ORS684 and ORS1110 can be effectively utilized for marker assisted breeding programme to develop the sunflower lines resistant to powdery mildew.

QTL Mining and Validation of Grain Nutritional Quality Characters in Rice (Oryza sativa L.) Using Two Introgression Line Populations

Nutritional grain quality is mainly influenced by the protein fraction content and grain protein content. Quantitative trait loci (QTL) mining for five traits, about 245 and 284 BC3F3 individual families of two introgression line (IL) populations were derived from Kongyu 131/Cypress (population-I) and Kongyu 131/Vary Tarva Osla (population-II), respectively. A genetic linkage map was developed using 127 simple sequence repeat (SSR) markers in population-I and 119 SSR markers in population-II. In total, 20 and 5 QTLs were detected in population-I and population-II, respectively. About twenty QTLs were mapped in population-I: five QTLs for albumin, seven QTLs for globulin, six QTLs for prolamin, one QTL for glutelin, and one QTL for grain protein content. In total, five QTLs were mapped in population-II: one QTL for albumin and four QTLs for grain protein content. Out of 25 QTLs, 19 QTLs exhibit co-localization with the previously reported QTLs. QTL-like qGPC7.3 was delineated for total protein content. This QTL was derived from population-I and was successfully validated in NILs (near-isogenic lines). The grain protein phenotype showed a significant variation between two NILs. This investigation serves as groundwork for additional cloning of nutritional quality-related genes in rice grains.

Multivariate analysis and genetic dissection of staygreen and stem reserve mobilisation under combined drought and heat stress in wheat (Triticum aestivum L.).

Introduction: Abiotic stresses significantly reduce crop yield by adversely affecting many physio-biochemical processes. Several physiological traits have been targeted and improved for yield enhancement in limiting environmental conditions. Amongst them, staygreen and stem reserve mobilisation are two important mutually exclusive traits contributing to grain filling under drought and heat stress in wheat. Henceforth, the present study was carried out to identify the QTLs governing these traits and to identify the superiors' lines through multi-trait genotype-ideotype distance index (MGIDI) Methods: A mapping population consisting of 166 recombinant inbred lines (RILs) developed from a cross between HD3086 and HI1500 was utilized in this study. The experiment was laid down in alpha lattice design in four environmental conditions viz. Control, drought, heat and combined stress (heat and drought). Genotyping of parents and RILs was carried out with 35K Axiom® array (Wheat breeder array). Results and Discussion: Medium to high heritability with a moderate to high correlation between traits was observed. Principal component analysis (PCA) was performed to derive latent variables in the original set of traits and the relationship of these traits with latent variables.From this study, 14 QTLs were identified, out of which 11, 2, and 1 for soil plant analysis development (SPAD) value, leaf senescence rate (LSR), and stem reserve mobilisation efficiency (SRE) respectively. Quantitative trait loci (QTLs) for SPAD value harbored various genes like Dirigent protein 6-like, Protein FATTY ACID EXPORT 3, glucan synthase-3 and Ubiquitin carboxyl-terminal hydrolase, whereas QTLs for LSR were found to contain various genes like aspartyl protease family protein, potassium transporter, inositol-tetrakisphosphate 1-kinase, and DNA polymerase epsilon subunit D-like. Furthermore, the chromosomal region for SRE was found to be associated with serine-threonine protein kinase. Serine-threonine protein kinases are involved in many signaling networks such as ABA mediated ROS signaling and acclimation to environmental stimuli. After the validation of QTLs in multilocation trials, these QTLs can be used for marker-assisted selection (MAS) in breeding programs.

Mapping and validation of quantitative trait loci associated with dorsal aleurone thickness in rice (Oryza sativa)

Mapping of QTLs for dorsal aleurone thickness (DAT) was performed using chromosome segment substitution lines in rice. Three QTLs, qDAT3.1, qDAT3.2, and qDAT7.1, were detected in multiple environments. As a specified endosperm cell type, the aleurone has an abundance of various nutrients. Increasing the number of aleurone layers is a practicable way of developing highly nutritious cereals. Identifying genes that can increase aleurone thickness is useful for the breeding of aleurone traits to improve the nutritional and health values of rice. Here, we found that iodine staining could efficiently distinguish the aleurone layers, which revealed great variation of the aleurone thickness in rice, especially at the dorsal side of the seed. Therefore, we used a population of chromosome segmental substitution lines (CSSLs) derived from Koshihikari and Nona Bokra for quantitative trait locus (QTL) analysis of the dorsal aleurone thickness (DAT). Three QTLs, qDAT3.1, qDAT3.2, and qDAT7.1, were detected in multiple seasons. Among these, qDAT3.2 colocalizes with Hd6 and Hd16, two QTLs previously identified to regulate the heading date of Koshihikari, explaining the negative correlation between the DAT and days to heading (DTH) in rice. We also provide evidence that early-heading ensures the filling of rice seed under a relatively high temperature to promote aleurone thickening. qDAT7.1, the most stable QTL expressed in different environments, functions independently from heading date. Although Nona Bokra has a lower DAT, its qDAT7.1 allele significantly increased DAT in rice, which was further validated using two near-isogenic lines (NILs). These findings pave the way for further gene cloning of aleurone-related QTLs and may aid the development of highly nutritious rice.

Identification and validation of quantitative trait loci for fertile spikelet number per spike and grain number per fertile spikelet in bread wheat (Triticum aestivum L.)

A major and stable QTL for fertile spikelet number per spike and grain number per fertile spikelet identified in a 4.96-Mb interval on chromosome 2A was validated in different genetic backgrounds. Fertile spikelet number per spike (FSN) and grain number per fertile spikelet (GNFS) contribute greatly to wheat yield improvement. To detect quantitative trait loci (QTL) associated with FSN and GNFS, we used a recombinant inbred line population crossed by Zhongkemai 13F10 and Chuanmai 42 in eight environments. Two Genomic regions associated with FSN were detected on chromosomes 2A and 6A using bulked segregant exome sequencing analysis. After the genetic linkage maps were constructed, four QTL QFsn.cib-2A, QFsn.cib-6A, QGnfs.cib-2A and QGnfs.cib-6A were identified in three or more environments. Among them, two major QTL QFsn.cib-2A (LOD = 4.67-9.34, PVE = 6.66-13.05%) and QGnfs.cib-2A (LOD = 5.27-11.68, PVE = 7.95-16.71%) were detected in seven and six environments, respectively. They were co-located in the same region, namely QFsn/Gnfs.cib-2A. The developed linked Kompetitive Allele Specific PCR (KASP) markers further validated this QTL in a different genetic background. QFsn/Gnfs.cib-2A showed pleiotropic effects on grain number per spike (GNS) and spike compactness (SC), and had no effect on grain weight. Since QFsn/Gnfs.cib-2A might be a new locus, it and the developed KASP markers can be used in wheat breeding. According to haplotype analysis, QFsn/Gnfs.cib-2A was identified as a target of artificial selection during wheat improvement. Based on haplotype analysis, sequence differences, spatiotemporal expression patterns, and gene annotation, the potential candidate genes for QFsn/Gnfs.cib-2A were predicted. These results provide valuable information for fine mapping and cloning gene(s) underlying QFsn/Gnfs.cib-2A.

Elucidation and characterization of QTLs for Russet formation on apple fruit in ‘Honeycrisp’-derived breeding germplasm

Russet formation in apples (Malus domestica Borkh.) is a superficial skin disorder that detracts from fruit appearance and therefore is a breeding target in many apple scion breeding programs. Previous studies have reported quantitative trait loci (QTLs) for russet formation, but these results have been limited to a narrow genetic background, which reduces breeding utility. In this study, 14 breeding parents were represented in 13-full sib families, nine of which were ‘Honeycrisp’-derived while the other four were derived from ‘Minneiska’, an offspring of ‘Honeycrisp.’ Two previously reported QTLs, on linkage group (LG) 2 and LG6, were confirmed and characterized for the first time in ‘Honeycrisp’-derived germplasm. Five other putative QTLs were also detected. The QTLs characterized in this study could be targeted for DNA test development to be used in marker-assisted selection. Non-additive interactions were observed at and between the LG2 and LG6 QTLs. Future work should include validation of QTL and haplotype effects in other germplasm, development of trait-predictive DNA tests for use in apple breeding programs, and exploration of genome-wide prediction models for russet formation.

Identification and validation of quantitative trait loci for chlorophyll content of flag leaf in wheat under different phosphorus treatments.

In wheat, the leaf chlorophyll content in flag leaves is closely related to the degree of phosphorus stress. Identifying major genes/loci associated with chlorophyll content in flag leaves under different phosphorus conditions is critical for breeding wheat varieties resistant to low phosphorus (P). Under normal, medium, and low phosphorus conditions, the chlorophyll content of flag leaves was investigated by a double haploid (DH) population derived from a cross between two popular wheat varieties Jinmai 47 and Jinmai 84, at different grain filling stages. Chlorophyll content of the DH population and parents decreased gradually during the S1 to the S3 stages and rapidly at the S4 stage. At the S4 stage, the chlorophyll content of the DH population under low phosphorus conditions was significantly lower than under normal phosphate conditions. Using a wheat 15K single-nucleotide polymorphism (SNP) panel, a total of 157 QTLs were found to be associated with chlorophyll content in flag leaf and were identified under three phosphorus conditions. The phenotypic variation explained (PVE) ranged from 3.07 to 31.66%. Under three different phosphorus conditions, 36, 30, and 48 QTLs for chlorophyll content were identified, respectively. Six major QTLs Qchl.saw-2B.1, Qchl.saw-3B.1, Qchl.saw-4D.1, Qchl.saw-4D.2, Qchl.saw-5A.9 and Qchl.saw-6A.4 could be detected under multiple phosphorus conditions in which Qchl.saw-4D.1, Qchl.saw-4D.2, and Qchl.saw-6A.4 were revealed to be novel major QTLs. Moreover, the closely linked SNP markers of Qchl.saw-4D.1 and Qchl.saw-4D.2 were validated as KASP markers in a DH population sharing the common parent Jinmai 84, showed extreme significance (P <0.01) in more than three environments under different phosphorus conditions, which has the potential to be utilized in molecular marker-assisted breeding for low phosphorus tolerance in wheat.

Mapping and validation of major and stable QTL for flag leaf size from tetraploid wheat.

The flag leaf is an important photosynthetic organ of wheat (Triticum aestivum L.). Appropriate flag leaf size can effectively increase grain yield. In this study, a tetraploid wheat population of recombinant inbred lines (RILs) and a genetic map constructed based on a wheat 55K single-nucleotide polymorphism (SNP) array were used to identify quantitative trait loci (QTL) for flag leaf length (FLL), flag leaf width (FLW), flag leaf area (FLA), and the flag leaf length/width ratio (FLR). A novel and major interval flanked by markers AX-111633224 and AX-109317229 was identified. This interval includes QTL for FLL (QFll.sau-AM-4B.2), for FLW (QFlw.sau-AM-4B.4), for FLA (QFla.sau-AM-4B), and for FLR (QFlr.sau-AM-4B). Based on the genotypes of the closely linked KASP (Kompetitive allele-specific polymerase chain reaction [PCR]) marker (KASP-AX-108756198), QFlw.sau-AM-4B.4 and QFla.sau-AM-4B were successfully verified in two F3 populations with different genetic backgrounds. Genetic associations between flag leaf-related traits and other agronomic traits were detected and analyzed. Four genes in this interval were likely involved in the growth and development of the flag leaf size. In conclusion, this study provides clues for excavating genes related to flag leaf size and breeding variety with ideal plant structure.

Identification and validation of major QTL for grain size and weight in bread wheat (Triticum aestivum L.)

Grain size and weight are key components of wheat yield. Exploitation of major underlying quantitative trait loci (QTL) can improve yield potential in wheat breeding. A recombinant inbred line (RIL) population was constructed to detect QTL for thousand-grain weight (TGW), grain length (GL) and grain width (GW) across eight environments. Genomic regions associated with grain size and grain weight were identified on chromosomes 4A and 6A using bulked segregant exome sequencing (BSE-Seq) analysis. After constructing genetic maps, six major QTL detected in at least four individual environments and in best linear unbiased estimator (BLUE) datasets, explained 7.50%–23.45% of the phenotypic variation. Except for QGl.cib-4A, the other five QTL were co-located in two regions, namely QTgw/Gw.cib-4A and QTgw/Gw/Gl.cib-6A. Interactions of these QTL were analyzed. Unlike QTgw/Gw/Gl.cib-6A, QTgw/Gw.cib-4A and QGl.cib-4A had no effect on grain number per spike (GNS). The QTL were validated in a second cross using Kompetitive Allele Specific PCR (KASP) markers. Since QTgw/Gw.cib-4A was probably a novel locus, it and the KASP markers reported here can be used in wheat breeding. TraesCS4A03G0191200 was predicted to be potential candidate gene for QTgw/Gw.cib-4A based on the sequence differences, spatiotemporal expression patterns, gene annotation and haplotype analysis. Our findings will be useful for fine mapping and for marker-assisted selection in wheat grain yield improvement.

Identification and validation of quantitative trait loci for a parameter associated with nitrogen partitioning to grain using a population derived from japonica- and indica-type cultivars of rice (Oryza sativa L.)

ABSTRACT Nitrogen (N) partitioning to grain affects productivity and grain quality in rice. The objectives of this study were to clarify the difference between the rice cultivars ‘Momiroman’ (japonica) and ‘Takanari’ (indica) in the relation between the amount of total N in aboveground parts per grain dry weight (TNA/GW) and grain N concentration (GNC), to identify quantitative trait loci (QTLs) associated with the coefficient characterizing this relation by using populations derived from a cross between Momiroman and Takanari, and to verify the effects of the detected QTLs by using near-isogenic lines (NILs). We used 156 F2 plants grown under high N in 2015 and 156 F3 lines grown under high or low N in 2016, and determined N concentrations and contents in grain and aboveground vegetative parts. We found a logarithmic relation between GNC and TNA/GW in Momiroman and Takanari. The regression coefficient (A) was higher in Takanari. Under each N condition, A was calculated for each population and QTL analysis was performed. QTLs for A were detected on chromosomes (Chrs.) 6 and 10 in all conditions; the Takanari alleles of both QTLs increased the value. NILs with the Takanari allele in each region had higher A than NILs with the Momiroman alleles. We conclude that the QTLs for A are associated with N partitioning to grain in rice. Abbreviations: GNA, amount of N in grain; GNC, grain N concentration; GW, grain dry weight; TNA, amount of N in aboveground parts

Identification and Validation of Quantitative Trait Loci for Grain Size in Bread Wheat (Triticum aestivum L.)

Grain width (GW) and grain length (GL) are crucial components affecting grain weight. Dissection of their genetic control is essential for improving yield potential in wheat breeding. Yangmai 12 (YM12) and Yanzhan 1 (YZ1) are two elite cultivars released in the Middle and Lower Yangtze Valleys Wheat Zone (MLYVWZ) and the Yellow-Huai River Valleys Wheat Zone (YRVWZ), respectively. One biparental population derived from YM12/YZ1 cross was employed to perform QTL mapping based on the data from four environments over two years to detect quantitative trait loci (QTL) for GW and GL. A total of eight QTL were identified on chromosomes 1B, 2D, 3B, 4B, 5A, and 6B. Notably, QGW.yz.2D was co-located with QGL.yz.2D, and QGW.yz.4B was co-located with QGL.yz.4B, respectively. QGW.yz.2D and QGL.yz.2D, with the increasing GW/GL allele from YZ1, explained 12.36–18.27% and 13.69–26.53% of the phenotypic variations for GW and GL, respectively. QGW.yz.4B and QGL.yz.4B, with the increasing GW/GL allele from YM12, explained 10.34–11.95% and 10.35–16.04% of the phenotypic variation for GW and GL, respectively. QGL.yz.5A, with the increasing GL allele from YM12, explained 10.04–12.48% of the phenotypic variation for GL. Moreover, the positive alleles of these three QTL regions could significantly increase thousand-grain weight, and QGW.yz.4B/QGL.yz.4B and QGL.yz.5A did not show significant negative effects on grain number per spike. QGL.yz.2D, QGW.yz.4B/QGL.yz.4B, and QGL.yz.5A have not been reported. These three QTL regions were then further validated using Kompetitive Allele-Specific PCR (KASP) markers in 159 wheat cultivars/lines from MLYVWZ and YRVWZ. Combining the positive alleles of the major QTL significantly increased GW and GL. Eleven candidate genes associated with encoding ethylene-responsive transcription factor, oleosin, osmotin protein, and thaumatin protein were identified. Three major QTL and KASP markers reported here will be helpful in developing new wheat cultivars with high and stable yields.

Molecular mapping and validation of quantitative trait loci for spike-related traits and plant height in wheat

<p id="C3">Spike-related traits and plant height are important target traits in wheat breeding. In the present study, a population of 198 recombinant inbred lines (RILs) derived from the cross between a CIMMYT wheat line C615 and Yangmai 13 (YM13) was constructed, followed by genotyping with Wheat 90K SNP array and phenotyping of spike-related traits and plant height in three environments to excavate QTLs (quantitative trait loci) for these traits. Using composite interval mapping method, we identified one QTL for total spikelet number per spike (TSS), two QTLs for spike length (SL), two QTLs for spikelet compactness (SC), and three QTLs for plant height (PH). <italic>QSN.yaas-3B</italic> and <italic>QPH.yaas-3B</italic> overlapped on the chromosome 3B. <italic>QSL.yaas-5A</italic>, <italic>QSC.yaas-5A</italic> and <italic>QPH.yaas-5A</italic> overlapped on the chromosome 5A. <italic>QSL.yaas-6A</italic> and <italic>QSC.yaas-6A</italic> overlapped on the chromosome 6A. <italic>QSN.yaas-3B</italic>/<italic>QPH.yaas-3B</italic> and <italic>QSL.yaas-6A</italic>/<italic>QSC.yaas-6A</italic> had not been reported yet, and were likely to be novel loci. The SNP marker closely linked to <italic>QSL.yaas-5A</italic>/<italic>QSC.yaas-5A</italic>/<italic>QPH.yaas-5A</italic> was then converted into one Kompetitive Allele Specific PCR (KASP) marker (QC615-5A-KASP), and validated in a panel of 105 wheat lines. The results would be useful for improvement of yield related traits in wheat breeding.

Identification and validation of QTLs for tuber quality related traits in greater yam (Dioscorea alata L.)

Two Dioscorea alata populations were generated by hand pollination between contrasted diploid genitors. Population A (74F × Kabusa) was composed of 121 progenies while population B (74F × 14M) involved 193 progenies. These two populations were assessed over two consecutive years regarding important tuber quality traits. Analysis of variance showed that the genotype had the greatest influence on the phenotypic scores. Also for some traits, effect of the year_replicate was strong. The heritabilities of most traits were high. Based on these data and a reference high-density genetic map of greater yam, a total of 34 quantitative trait loci (QTLs) were detected on 8 of the 20 yam chromosomes. They corresponded to five of each of the following traits: tuber size, shape regularity, tubercular roots, skin texture, tuber flesh oxidation, six for oxidation ratio and three for flesh colour. The fraction of total phenotypic variance attributable to a single QTL ranged from 11.1 to 43.5%. We detected significant correlations between traits and QTL colocalizations that were consistent with these correlations. A majority of QTLs (62%) were found on linkage group LG16, indicating that this chromosome could play a major role in genetic control of the investigated traits. In addition, an inversion involving this chromosome was detected in the Kabusa male. Nine QTLs were validated on a diversity panel, including three for tuber size, three for shape regularity, two for skin texture and one for tubercular roots. The approximate physical localization of validated QTLs allowed the identification of various candidates genes. The validated QTLs should be useful for breeding programs using marker-assisted selection to improve yam tuber quality.

Identification and Validation of Quantitative Trait Loci for Wheat Dwarf Virus Resistance in Wheat (Triticum spp.).

Wheat dwarf virus (WDV) is transmitted by the leafhopper Psammotettix alienus. As a major pathogen in wheat and other cereals, WDV causes high yield losses in many European countries. Due to climate change, insect-transmitted viruses will become more important and the restrictions in the use of insecticides efficient against P. alienus renders growing of WDV resistant/tolerant varieties the only effective strategy to control WDV. So far, there is little information about the possible sources of resistance and no known information about the genome regions responsible for the resistance. In a screening for WDV resistance using artificial inoculation in gauze houses, a panel of 500 wheat accessions including cultivars, gene bank accessions, and wild relatives of wheat was phenotyped for virus titer, infection rate, as well as plant height and yield parameters relative to healthy controls of the same genotype. Additionally, 85 T. aestivum-Ae. tauschii intogression lines were tested for WDV resistance in the greenhouse. A subset of 250 hexaploid wheat accessions was genotyped with the 15k iSelect SNP Chip. By genome-wide association study (GWAS), the quantitative trait loci (QTL) for partial WDV resistance were identified. Within these studies, one cultivar was identified showing an average infection rate of only 5.7%. By analyzing single seed descent (SSD) and doubled haploid (DH) populations comprising 153 and 314 individuals for WDV resistance and by genotyping these with the 25k iSelect SNP Chip, QTL for yield per plant, thousand-grain weight, and relative virus titer were validated on chromosomes 1B, 2B, 3B, 4B, 4A, 5A, 6A, and 7A. These results will be the basis for marker-assisted selection for WDV resistance to replacing the laborious, time-consuming, and technically challenging phenotyping with WDV bearing leafhoppers.

Identification and validation of quantitative trait loci for the functional stay green trait in common wheat (Triticum aestivum L.) via high-density SNP-based genotyping.

This study identified QTLs associated with the functional stay green trait by a high-density genetic map. Two large effect QTLs, QSg.sau-2B.1 and QSg.sau-6A.2, were identified in multiple years and one of them was successfully validated. The functional stay green phenotype enables wheat to acclimate to stressful environments and prolongs the effectiveness of photosynthesis during the end-of-crop season. Despite the fact that stay green mutants in wheat have been reported, our knowledge of loci for the functional stay green trait remains limited. In this study, an RIL population containing 371 lines genotyped using the Wheat55K SNP array was used to map QTLs controlling the functional stay green trait in multiple years. In total, 21 and 19 QTLs were mapped using the BIP or MET modules of the ICIM method, respectively. Among them, two QTLs, QSg.sau-2B.1 and QSg.sau-6A.2, were considered large effect QTLs for the stay green trait and explained 11.43% and 15.27% of phenotypic variation on average, respectively. Two KASP markers were developed and tightly linked to QSg.sau-2B.1 and QSg.sau-6A.2, respectively, and the genetic effects of different genotypes in the RIL population were successfully confirmed. QSg.sau-2B.1 was also validated by linked KASP marker in different genetic backgrounds. QSg.sau-2B.1 and QSg.sau-6A.2 may influence heredity of the stay green trait and also exhibited a positive effect on the grain filling content. In the interval where QSg.sau-2B.1 and QSg.sau-6A.2 were located on the Chinese Spring and T. turgidum ssp. dicoccoides reference genomes, several genes associated with the leaf senescence process were identified. Altogether, our results identified two QTLs associated with the functional stay green trait and will be useful for the fine mapping and cloning of genes for stay green in the future.

Identification of quantitative trait loci associated with upper temperature tolerance in turbot, Scophthalmus maximus

Temperature tolerance is an important trait from both an economic and evolutionary perspective in fish. Because of difficulties with measurements, genome-wide selection using quantitative trait loci (QTLs) affecting Upper temperature tolerance may be an alternative for genetic improvement. Turbot Scophthalmus maximus (L.) is a cold-water marine fish with high economic value in Europe and Asia. The genetic bases of upper temperature tolerance (UTTs) traits have been rarely studied. In this study, we constructed a genetic linkage map of turbot using simple sequence repeats (SSRs) and single nucleotide polymorphism (SNP) markers. A total of 190 SSR and 8,123 SNP were assigned to 22 linkage groups (LGs) of a consensus map, which spanned 3,648.29 cM of the turbot genome, with an average interval of 0.44 cM. Moreover, we re-anchored genome sequences, allowing 93.8% physical sequences to be clustered into 22 turbot pseudo-chromosomes. A high synteny was observed between two assemblies from the literature. QTL mapping and validation analysis identified thirteen QLTs which are major effect QTLs, of these, 206 linked SNP loci, and two linked SSR loci were considered to have significant QTL effects. Association analysis for UTTs with 129 QTL markers was performed for different families, results showed that eight SNP loci were significantly correlated with UTT, which markers could be helpful in selecting thermal tolerant breeds of turbot. 1,363 gene sequences were genomically annotated, and 26 QTL markers were annotated. We believe these genes could be valuable candidates affecting high temperatures, providing valuable genomic resources for the study of genetic mechanisms regulating thermal stress. Similarly, they may be used in marker-assisted selection (MAS) programs to improve turbot performance.

QTL Mapping and Validation for Kernel Area and Circumference in Common Wheat via High-Density SNP-Based Genotyping.

As an important component, 1,000 kernel weight (TKW) plays a significant role in the formation of yield traits of wheat. Kernel size is significantly positively correlated to TKW. Although numerous loci for kernel size in wheat have been reported, our knowledge on loci for kernel area (KA) and kernel circumference (KC) remains limited. In the present study, a recombinant inbred lines (RIL) population containing 371 lines genotyped using the Wheat55K SNP array was used to map quantitative trait loci (QTLs) controlling the KA and KC in multiple environments. A total of 54 and 44 QTLs were mapped by using the biparental population or multienvironment trial module of the inclusive composite interval mapping method, respectively. Twenty-two QTLs were considered major QTLs. BLAST analysis showed that major and stable QTLs QKc.sau-6A.1 (23.12–31.64 cM on 6A) for KC and QKa.sau-6A.2 (66.00–66.57 cM on 6A) for KA were likely novel QTLs, which explained 22.25 and 20.34% of the phenotypic variation on average in the 3 year experiments, respectively. Two Kompetitive allele-specific PCR (KASP) markers, KASP-AX-109894590 and KASP-AX-109380327, were developed and tightly linked to QKc.sau-6A.1 and QKa.sau-6A.2, respectively, and the genetic effects of the different genotypes in the RIL population were successfully confirmed. Furthermore, in the interval where QKa.sau-6A.2 was located on Chinese Spring and T. Turgidum ssp. dicoccoides reference genomes, only 11 genes were found. In addition, digenic epistatic QTLs also showed a significant influence on KC and KA. Altogether, the results revealed the genetic basis of KA and KC and will be useful for the marker-assisted selection of lines with different kernel sizes, laying the foundation for the fine mapping and cloning of the gene(s) underlying the stable QTLs detected in this study.

Identification and Validation of QTLs for Yield and Yield Components under Long-Term Salt Stress Using IR64 CSSLs in the Genetic Background of Koshihikari and Their Backcross Progenies

Unraveling the complex genetic bases and mechanisms underlying salt tolerance is of great importance for developing salt-tolerant varieties. In this study, we evaluated 42 chromosome segment substitution lines (CSSLs) carrying chromosome segments from IR64 on the genetic background of Koshihikari under salt stress. Two CSSLs, SL2007 and SL2038, produced higher plant dry weight and grain yield than did Koshihikari under the stress condition. These CSSLs also showed lower Na+ and Cl− accumulation in the leaf and whole plant at the full heading stage, which might be related to the higher grain yield and yield components. To understand the genetic control of its grain yield and yield components, a SL2007/Koshihikari F2 population was generated for quantitative trait locus (QTL) analysis. Six QTLs for grain yield and yield-related traits were detected on chromosome 2. Using near-isogenic lines (NILs) from a SL2007/Koshihikari F5 population, qSTGY2.2 was delimited to a 2.5 Mb region and novel qSTPN2 was delimited to a 0.6 Mb region. We also detected a novel QTL, qSTGF2, for grain filling, which was considered an important contributor to grain yield under salt stress in this CSSL. Our results provide insights into mechanisms conferring grain yield under salinity stress and new genetic resources for cloning and breeding.

Mapping and validation of quantitative trait loci that confer resistance to rice black-streaked dwarf virus disease in rice (Oryza sativa)

Rice black-streaked dwarf virus (RBSDV) disease is one of the most destructive viral diseases that threatens rice production in China. Breeding of resistant cultivars through multi-gene pyramiding is considered to be an effective way to control the disease, but few resistance genes have been characterized to date. In the present study, we identified T1012, a BC2F6 line from a cross of the japonica variety ‘Wuyujing3’ (recipient) and the indica variety ‘Dular’ (donor), that had improved resistance to RBSDV disease in a field test, and 140 backcross inbred lines (BILs) derived from a cross between T1012 and ‘Wuyujing3’ were developed using marker-assisted selection. Genetic analysis showed that the resistance of T1012 to RBSDV disease was controlled by quantitative trait loci (QTLs). Two QTLs for RBSDV disease resistance located on chromosomes 1 and 4, qRBSDV-1 and qRBSDV-4, were identified, and qRBSDV-4 was repeatedly detected in two environments. Compared to ‘Wuyujing3’, the line containing only the substitution segment covering qRBSDV-4 exhibited significantly decreased disease incidence, indicating that qRBSDV-4 is a reliable resistance QTL with a high breeding value. Furthermore, two linked QTLs, qRBSDV-4-1 and qRBSDV-4-2, were identified within the interval containing qRBSDV-4. The QTLs identified here will provide a useful resource for breeding RBSDV-resistant rice cultivars through marker-assisted selection and establish a foundation for the cloning of RBSDV disease resistance genes.