Chromosome 2DL Research Articles

Fine mapping of two recessive genes TaFLA1 and TaSPL8 controlling flag leaf angle in bread wheat

Flag leaf angle is one of the key target traits in high yield wheat breeding. A smaller flag leaf angle reduces shading and enables plants to grow at a higher density, which increases yield. Here we identified a mutant, je0407, with an 84.34%–89.35% smaller flag leaf angle compared with the wild type. The mutant also had an abnormal lamina joint and no ligule or auricle. Genetic analysis indicated that the ligule was controlled by two recessive genes, which were mapped to chromosomes 2AS and 2DL. The mutant allele on chromosome 2AS was named Tafla1b, and it was fine mapped to a 1 Mb physical interval. The mutant allele on chr. 2DL was identified as Taspl8b, a novel allele of TaSPL8 with a missense mutation in the second exon, which was used to develop a cleaved amplified polymorphic sequence marker. F3 and F4 lines derived from crosses between Jing411 and je0407 were genotyped to investigate interactions between the Tafla1b and Taspl8b alleles. Plants with the Tafla1b/Taspl8a genotype had 58.41%–82.76% smaller flag leaf angles, 6.4%–24.9% shorter spikes, and a greater spikelet density (0.382 more spikelets per cm) compared with the wild type. Plants with the Tafla1a/Taspl8b genotype had 52.62%–82.24% smaller flag leaf angles and no differences in plant height or spikelet density compared with the wild type. Tafla1b/Taspl8b plants produced erect leaves with an abnormal lamina joint. The two alleles had dosage effects on ligule formation and flag leaf angle, but no significant effect on thousand-grain weight. The mutant alleles provide novel resources for improvement of wheat plant architecture.

Identification of a major QTL conferring resistance to wheat yellow mosaic virus derived from the winter wheat 'Hokkai 240' on chromosome 2AS.

Wheat yellow mosaic disease is a soilborne disease caused by wheat yellow mosaic virus (WYMV). Symptoms include yellow mosaic coloring of leaves, stunting, and growth inhibition. Here we conducted a detailed analysis of resistance to this virus in winter wheat 'Hokkai 240' by carrying out inoculation tests of WYMV and conducting field tests. The resistance level observed in 'Hokkai 240' was compared with those in varieties harboring known resistance genes. In the inoculation tests, 'Hokkai 240' showed resistance to WYMV Pathotypes I and II and partial resistance to Pathotype III. This result was contrary to the sensitive responses to the three pathotypes exhibited by the variety harboring resistance gene on chromosome 2DL. In fields infected with WYMV Pathotypes II and III, 'Hokkai 240' plants exhibited few disease symptoms and little proliferation of the virus. By analyzing the quantitative trait loci (QTLs) in recombinant inbred lines from a cross between 'Hokkai 240' and 'Nanbukomugi', a single major QTL, Q.Ymhk, from 'Hokkai 240', which had significant effects on Pathotypes II and III of the virus, was detected in the proximity of snp4212 and snp4215 mapped on chromosome 2AS. These results indicate that Q.Ymhk may be useful for developing broad resistance to WYMV in wheat breeding programs.

Molecular identification and validation of four stable QTL for adult-plant resistance to powdery mildew in Chinese wheat cultivar Bainong 64.

Four stable QTL for adult-plant resistance (APR) to powdery mildew were identified on chromosome arms 1DL, 2BS, 2DL, and 6BL in the widely grown Chinese wheat cultivar Bainong 64. These QTL had no effect on response to stripe rust or leaf rust. Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a devastating fungal disease. Seedlings of Chinese wheat Bainong 64 are susceptible to Bgt, but adult plants have maintained resistance since it was released in 1996. A population of 171 recombinant inbred lines (RILs) developed from cross Jingshuang 16/Bainong 64 (JS16/BN64) was used to dissect genetic components of powdery mildew resistance. A genetic map comprising 5383 polymorphic markers was constructed using the 15K SNP chip and kompetitive allele-specific PCR (KASP) markers. Composite interval mapping identified four stable QTL with favorable alleles all from BN64 on chromosome arms 1DL, 2BS, 2DL, and 6BL in at least four environments. They accounted for 8.3%, 13.8%, 14.4%, and 9.0% of the total phenotypic variation explained (PVE) in maximum, respectively. QPmjbr.caas-1DL, situated about 22Mb from centromere, is probably a new QTL. QPmjbr.caas-2DL located near the end of arm 2DL and explained the largest PVE. Using genetic maps populated with KASP markers, QPmjbr.caas-2BS and QPmjbr.caas-6BL were fine mapped to a 1.8cM genetic intervals spanning 13.6Mb (76.0-89.6Mb) and 1.7cM and 4.9Mb (659.9-664.8Mb), respectively. The four QTL independent of stripe rust and leaf rust resistance were validated for powdery mildew resistance in another RIL population related to BN64 and a cultivar panel using representative KASP markers. Since BN64 has been a leading cultivar and an important breeding parent in China, the QTL and markers reported in this study will be useful for marker-assisted selection of APR.

Molecular characterization of stable QTL and putative candidate genes for grain zinc and iron concentrations in two related wheat populations.

Major QTL for grain zinc and iron concentrations were identified on the long arm of chromosomes 2D and 6D. Gene-based KASP markers were developed for putative candidate genes TaIPK1-2D and TaNAS10-6D. Micronutrient malnutrition is one of the most common public health problems in the world. Biofortification, the most attractive and sustainable solution to surmount malnutrition requires the development of micronutrient enriched new crop cultivars. In this study, two recombinant inbred line (RIL) populations, ZM175/XY60 and ZM175/LX987, were used to identify QTL for grain zinc concentration (GZnC), grain iron concentration (GFeC) and thousand grain weight (TGW). Eight QTL for GZnC, six QTL for GFeC and five QTL for TGW were detected. Three QTL on chromosomes 2DL and 4BS and chromosome 6A showed pleiotropic effects on all three traits. The 4BS and 6A QTL also increased plant height and might be Rht-B1a and Rht25a, respectively. The 2DL locus within a suppressed recombination region was identified in both RIL populations and the favorable allele simultaneously increasing GZnC, GFeC and TGW was contributed by XY60 and LX987. A QTL on chromosome 6DL associated only with GZnC was detected in ZM175/XY60 and was validated in JD8/AK58 RILs using kompetitive allele-specific PCR (KASP) marker K_AX-110119937. Both the 2DL and 6DL QTL were new loci for GZnC. Based on gene annotations, sequence variations and expression profiles, thephytic acid biosynthesis gene TaIPK1-2D and nicotianamine synthase gene TaNAS10-6D were predicted as candidate genes. Their gene-based KASP markers were developed and validated in a cultivar panel of 343 wheat accessions. This study investigated the genetic basis of GZnC and GFeC and provided valuable candidate genes and markers for breeding Zn- and Fe-enriched wheat.

DMC1 stabilizes crossovers at high and low temperatures during wheat meiosis.

Effective chromosome synapsis and crossover formation during meiosis are essential for fertility, especially in grain crops such as wheat. These processes function most efficiently in wheat at temperatures between 17-23 °C, although the genetic mechanisms for such temperature dependence are unknown. In a previously identified mutant of the hexaploid wheat reference variety 'Chinese Spring' lacking the long arm of chromosome 5D, exposure to low temperatures during meiosis resulted in asynapsis and crossover failure. In a second mutant (ttmei1), containing a 4 Mb deletion in chromosome 5DL, exposure to 13 °C led to similarly high levels of asynapsis and univalence. Moreover, exposure to 30 °C led to a significant, but less extreme effect on crossovers. Previously, we proposed that, of 41 genes deleted in this 4 Mb region, the major meiotic gene TaDMC1-D1 was the most likely candidate for preservation of synapsis and crossovers at low (and possibly high) temperatures. In the current study, using RNA-guided Cas9, we developed a new Chinese Spring CRISPR mutant, containing a 39 bp deletion in the 5D copy of DMC1, representing the first reported CRISPR-Cas9 targeted mutagenesis in Chinese Spring, and the first CRISPR mutant for DMC1 in wheat. In controlled environment experiments, wild-type Chinese Spring, CRISPR dmc1-D1 and backcrossed ttmei1 mutants were exposed to either high or low temperatures during the temperature-sensitive period from premeiotic interphase to early meiosis I. After 6-7 days at 13 °C, crossovers decreased by over 95% in the dmc1-D1 mutants, when compared with wild-type plants grown under the same conditions. After 24 hours at 30 °C, dmc1-D1 mutants exhibited a reduced number of crossovers and increased univalence, although these differences were less marked than at 13 °C. Similar results were obtained for ttmei1 mutants, although their scores were more variable, possibly reflecting higher levels of background mutation. These experiments confirm our previous hypothesis that DMC1-D1 is responsible for preservation of normal crossover formation at low and, to a certain extent, high temperatures. Given that reductions in crossovers have significant effects on grain yield, these results have important implications for wheat breeding, particularly in the face of climate change.

Genomic Regions Associated with Resistance to Three Rusts in CIMMYT Wheat Line "Mokue#1".

Understanding the genetic basis of rust resistance in elite CIMMYT wheat germplasm enhances breeding and deployment of durable resistance globally. "Mokue#1", released in 2023 in Pakistan as TARNAB Gandum-1, has exhibited high levels of resistance to stripe rust, leaf rust, and stem rust pathotypes present at multiple environments in Mexico and Kenya at different times. To determine the genetic basis of resistance, a F5 recombinant inbred line (RIL) mapping population consisting of 261 lines was developed and phenotyped for multiple years at field sites in Mexico and Kenya under the conditions of artificially created rust epidemics. DArTSeq genotyping was performed, and a linkage map was constructed using 7892 informative polymorphic markers. Composite interval mapping identified three significant and consistent loci contributed by Mokue: QLrYr.cim-1BL and QLrYr.cim-2AS on chromosome 1BL and 2AS, respectively associated with stripe rust and leaf rust resistance, and QLrSr.cim-2DS on chromosome 2DS for leaf rust and stem rust resistance. The QTL on 1BL was confirmed to be the Lr46/Yr29 locus, whereas the QTL on 2AS represented the Yr17/Lr37 region on the 2NS/2AS translocation. The QTL on 2DS was a unique locus conferring leaf rust resistance in Mexico and stem rust resistance in Kenya. In addition to these pleiotropic loci, four minor QTLs were also identified on chromosomes 2DL and 6BS associated with stripe rust, and 3AL and 6AS for stem rust, respectively, using the Kenya disease severity data. Significant decreases in disease severities were also demonstrated due to additive effects of QTLs when present in combinations.

Identification of stable quantitative trait loci underlying waterlogging tolerance post-anthesis in common wheat (Triticum aestivum)

Waterlogging is a growing threat to wheat production in high-rainfall areas. In this study, a doubled haploid (DH) population developed from a cross between Yangmai 16 (waterlogging-tolerant) and Zhongmai 895 (waterlogging-sensitive) was used to map quantitative trait loci (QTL) for waterlogging tolerance using a high-density 660K single-nucleotide polymorphism (SNP) array. Two experimental designs, waterlogging concrete tank (CT) and waterlogging plastic tank (PT), were used to simulate waterlogging during anthesis in five environments across three growing seasons. Waterlogging significantly decreased thousand-kernel weight (TKW) relative to non-waterlogged controls, although the degree varied across lines. Three QTL for waterlogging tolerance were identified on chromosomes 4AL, 5AS, and 7DL in at least two environments. All favorable alleles were contributed by the waterlogging-tolerant parent Yangmai 16. QWTC.caas-4AL exhibited pleiotropic effects on both enhancing waterlogging tolerance and decreasing plant height. Six high-confidence genes were annotated within the QTL interval. The combined effects of QWTC.caas-4AL and QWTC.caas-5AS greatly improved waterlogging tolerance, while the combined effects of all three identified QTL (QWTC.caas-4AL, QWTC.caas-5AS, and QWTC.caas-7DL) exhibited the most significant effect on waterlogging tolerance. Breeder-friendly kompetitive allele-specific PCR (KASP) markers (K_AX_111523809, K_AX_108971224, and K_AX_110553316) flanking the interval of QWTC.caas-4AL, QWTC.caas-5AS, and QWTC.caas-7DL were produced. These markers were tested in a collection of 240 wheat accessions, and three superior polymorphisms of the markers distributed over 67 elite cultivars in the test population, from the Chinese provinces of Jiangsu, Anhui, and Hubei. The three KASP markers could be used for marker-assisted selection (MAS) to improve waterlogging tolerance in wheat.

Identification of Novel QTLs/Defense Genes in Spring Wheat Germplasm Panel for Seedling and Adult Plant Resistance to Stem Rust and Their Validation Through KASP Marker Assays.

Stem rust is one of the major diseases threatening wheat production globally. To identify novel resistance quantitative trait loci (QTLs), we performed 35K Axiom Array SNP genotyping assays on an association mapping panel of 400 germplasm accessions, including Indian landraces, in conjunction with phenotyping for stem rust at seedling and adult plant stages. Association analyses using three genome wide association study (GWAS) models (CMLM, MLMM, and FarmCPU) revealed 20 reliable QTLs for seedling and adult plant resistance. Among these 20 QTLs, five QTLs were found consistent with three models, i.e., four QTLs on chromosome 2AL, 2BL, 2DL, and 3BL for seedling resistance and one QTL on chromosome 7DS for adult plant resistance. Further, we identified a total of 21 potential candidate genes underlying QTLs using gene ontology analysis, including a leucine rich repeat receptor (LRR) and P-loop nucleoside triphosphate hydrolase, which have a role in pathogen recognition and disease resistance. Furthermore, four QTLs (Qsr.nbpgr-3B_11, QSr.nbpgr-6AS_11, QSr.nbpgr-2AL_117-6, and QSr.nbpgr-7BS_APR) were validated through KASP located on chromosomes 3B, 6A, 2A, and 7B. Out of these QTLs, QSr.nbpgr-7BS_APR was identified as a novel QTL for stem rust resistance which has been found effective in both seedling as well as the adult plant stages. Identified novel genomic regions and validated QTLs have the potential to be deployed in wheat improvement programs to develop disease resistant varieties for stem rust and can diversify the genetic basis of resistance.

Mapping the quantitative field resistance to stripe rust in a hard winter wheat population “Overley” × “Overland”

AbstractStripe (yellow) rust, caused by Puccinia striiformis f. sp. tritici, is a devastating disease of wheat (Triticum aestivum) worldwide. In commercial production, stripe rust reduces grain quality, grain yield, and forage yield. This study was conducted to identify quantitative trait locus (QTL) associated with field resistance to stripe rust in hard winter wheat. Stripe rust infection type and severity were rated in recombinant inbred lines (RILs, n = 204) derived from a cross between hard red winter wheat cultivars “Overley” and “Overland” in replicated field trials in the Great Plains and Pacific Northwest. RILs (n = 184) were genotyped with reduced representation sequencing to produce single nucleotide polymorphism (SNP) markers from alignment to the “Chinese Spring” reference sequence, IWGSC v2.1, and from alignment to the reference sequence for “Jagger,” which is a parent of Overley. Genetic linkage maps were developed independently from each set of SNP markers. QTL analysis identified genomic regions on chromosome arms 2AS, 2BS, 2BL, and 2DL that were associated with stripe rust resistance using multi‐environment best linear unbiased predictors for stripe rust infection type and severity. Results for the two linkage maps were very similar. PCR‐based SNP marker assays associated with the QTL regions were developed to efficiently identify these genomic regions in breeding populations.

Mapping of leaf rust resistance genes in common wheat ‘Guinong08-6’

Wheat leaf rust (Puccinia triticina Eriks.), a devastating disease of wheat in the world, causes severe yield losses and therefore the development of resistant cultivars is very important. Here, a Chinese wheat line (Guinong08-6) showed adult-plant resistance against mixed fungal isolates of leaf rust, which is common in Guiyang region. It was crossed with a susceptible wheat line (Guinong19) to develop F1, F2, and F3 hybrids. Combined SSR and STS markers were used to map leaf rust resistance genes in Guinong08-6, and the resistance phenotype of Guinong08-6 was co-regulated by two complementary dominant genes, named LrGn08-6A and LrGn08-6B. LrGn08-6A was mapped to chromosome 2AS with markers URIC-LN2 and Xgpw2204, which flanked the gene at distances of 1.8 centimorgan (cM) and 14.83 cM, respectively. LrGn08-6B was mapped to chromosome 4DL with markers Xgpw342 and Xbarc93, which both flanked the gene at a distance of 26.57 cM. Genetic and molecular marker analyses demonstrated that LrGn08-6A, which was inherited from Aegilops ventricosa may be the resistance gene Lr37, while LrGn08-6B may be a newly discovered leaf rust resistance gene.

Relocation of Sr48 to Chromosome 2D Using an Alternative Mapping Population and Development of a Closely Linked Marker Using Diverse Molecular Technologies.

The Ug99-effective stem rust resistance gene Sr48 was mapped to chromosome 2A based on its repulsion linkage with Yr1 in an Arina/Forno recombinant inbred line (RIL) population. Attempts to identify markers closely linked to Sr48 using available genomic resources were futile. This study used an Arina/Cezanne F5:7 RIL population to identify markers closely linked with Sr48. Using the Arina/Cezanne DArTseq map, Sr48 was mapped on the short arm of chromosome 2D and it co-segregated with 12 markers. These DArTseq marker sequences were used for BlastN search to identify corresponding wheat chromosome survey sequence (CSS) contigs, and PCR-based markers were developed. Two simple sequence repeat (SSR) markers, sun590 and sun592, and two Kompetitive Allele-Specific PCR (KASP) markers were derived from the contig 2DS_5324961 that mapped distal to Sr48. Molecular cytogenetic analysis using sequential fluorescent in situ hybridization (FISH) and genomic in situ hybridization (GISH) identified a terminal translocation of chromosome 2A in chromosome 2DL of Forno. This translocation would have led to the formation of a quadrivalent involving chromosomes 2A and 2D in the Arina/Forno population, which would have exhibited pseudo-linkage between Sr48 and Yr1 in chromosome 2AL. Polymorphism of the closet marker sunKASP_239 among a set of 178 wheat genotypes suggested that this marker can be used for marker-assisted selection of Sr48.

QTL analysis identified two major all-internodes solidness loci from a completely solid-stemmed spring wheat line.

The culms of solid-stemmed wheat cultivars are filled with "pith" - a parenchymatous tissue largely composed of soft, spongy, and compact parenchyma cells. Breeding solid-stemmed cultivars is the most effective way to decrease the detrimental impact of wheat stem sawfly (WSS), Cephus cinctus Norton (Hymenoptera: Cephidae) on wheat production. Althougha major solid stem gene has been previously identified from durum wheat, it produces an intermediate level of stem solidness in common wheat which is insufficient to provide the required level of WSS resistance. The maximum resistance is achieved when stems are totally filled with pith. Thus, to identify a secondary source of solidness in common wheat, we developed three mapping populations from wheat cvs. Sadash, 'AAC Innova' and 'AAC Cameron', each crossed separatelywith P2711, a completely solid-stemmed hexaploid wheat breeding line. All populations were genotyped using either wheat 15K or 90K Infinium iSelect SNP Assay and high-density linkage maps were generated from individual populations along with consensus maps for chromosomes 3B and 3D from all populations. 'Sadash/P2711' and 'AAC Innova/P2711' populations were subjected to extensive phenotyping in ≥3 environments followed by quantitative trait loci (QTL) analyses using population-specific and consensus linkage maps. We identified two major solid stem QTLs in the distal regions of chromosome arms 3BL and 3DL in both populations in addition to several population-specific or common minor QTLs. Internode-specific QTL analyses detected both major QTLs of chromosomes 3B and 3D across internodes, from top to bottom of the stalk, but minor QTLs were largely detected in upper or middle internodes. Our results suggest that both major QTLs are sufficient to develop highly solid-stemmed cvs; however, the minor loci, which additively enhance the pith expression, can be coupled with major genes to achieve a complete solid stem phenotype in common wheat. Comparative and haplotype analyses showed that the 3B locus is homoeologous to 3D, the former being mapped to a 1.1 Mb genomic region. Major QTLs identified in this study can be incorporated in modern wheat cultivars to achieve maximum WSS resistance from high pith expression.

Detection of resistance loci to English grain aphid in common wheat

AbstractThe English grain aphid (Sitobion avenae F.) causes significant yield and quality losses in wheat (Triticum aestivum L.) worldwide, and the use of insect‐resistant wheat cultivars reduces the loss level. The present study aimed to identify quantitative trait loci (QTL) and perform putative candidate gene evolutionary analysis for English grain aphid resistance in 261 F2:5 recombinant inbred lines (RILs) derived from the cross Amigo/XN1376. The RIL population and parents were genotyped with the Wheat 55K single nucleotide polymorphism (SNP) array. A total of 14,694 polymorphic SNPs were used to construct a genetic linkage map. One major effect of QTL was detected in F2:5 RILs and was located on chromosome 3DL, explained 19.33% phenotypic variance, temporarily designated as QSa3.haust‐3D. It was located in a 2.73 Mb, flanked by markers AX‐109429351 and AX‐110834100, from 550.25 to 552.98 Mb according to Chinese Spring reference genome. This region contained 39 annotated genes. Eight putative candidate genes for resistance to English grain aphid were identified among these genes. The QTL with closely linked SNP markers could be useful for marker‐assisted selection for resistance to English grain aphid for wheat breeding.

Genetic dissection of lutein content in common wheat via association and linkage mapping.

Genetic architecture controlling grain lutein content of common wheat was investigated through an integration of genome-wide association study (GWAS) and linkage analysis. Putative candidate genes involved in carotenoid metabolism and regulation were identified, which provide a basis for gene cloning and development of nutrient-enriched wheat varieties through molecular breeding. Lutein, known as 'the eye vitamin', is an important component of wheat nutritional and end-use quality. However, the genetic manipulation of grain lutein content (LUC) in common wheat has not previously been well studied. Here, quantitative trait loci (QTL) associated with the LUC measured by high performance liquid chromatography (HPLC) were first identified by integrating a genome-wide association study (GWAS) and linkage mapping. A Chinese wheat mini-core collection (MCC) of 262 accessions and a doubled haploid (DH) population derived from Jinchun 7 and L1219 were genotyped using the 90K SNP array. A total of 124 significant marker-trait associations (MTAs) on all 21 wheat chromosomes except for 1A, 4D, and 5B that formed 58 QTL were detected. Among them, six stable QTL were identified on chromosomes 2AL, 2DS, 3BL, 3DL, 7AL, and 7BS. Meanwhile, three of the ten QTL identified in the DH population, QLuc.5A.1 and QLuc.5A.2 on chromosome 5AL and QLuc.6A.2 on 6AS, were stable and independently explained 5.58-10.86% of the phenotypic variation. The QLuc.6A.2 region colocalized with two MTAs identified by GWAS. Moreover, 71 carotenoid metabolism-related candidate genes were identified, and the allelic effects were analyzed in the MCC panel based on the 90K array. Results revealed that the genes CYP97A3 (Chr. 6B) and CCD1 (Chr. 5A) were significantly associated with LUC. Additionally, the gene PSY3 (QLuc.5A.1) and several candidate genes involved in the methylerythritol 4-phosphate (MEP) pathways colocalized with stable QTL regions. The present study provides potential targets for future functional gene exploration and molecular breeding in common wheat.

Molecular analyses in wheat and Aegilops tauschii reveal a new orthologue of the leaf rust resistance gene Lr19 on chromosome 7DL of Ae. tauschii

AbstractLeaf rust is one of the most devastating wheat diseases worldwide, to which many resistance genes have been successfully introgressed from wheat wild relatives. Though the Thinopyrum ponticum‐derived leaf rust resistance gene Lr19 is widely effective, and previous studies have shown its likely presence in Aegilops tauschii, no thorough investigation has been conducted to confirm this. The current study examined the presence of Lr19 in Ae. tauschii using molecular and bioinformatic analyses. Accordingly, the wheat cultivar Thatcher was used as a susceptible control, and the Lr19‐carrying genotypes; Thatcher + Lr19 (TcLr19) and Agatha were used as resistant controls. Phenotyping using pathotype CDHLQ accompanied with genotyping using two DNA markers verified previous results of the likely presence of an orthologue for Lr19 on chromosome 7DL of Ae. tauschii (AtLr19). Sequencing of the STS GB marker products from Ae. tauschii and TcLr19 showed 99% homology between the corresponding fragments, confirming the phenotyping and genotyping results. Both sequenced fragments matched to the melatonin biosynthesis gene, O‐methyltransferase‐2 (OMT2). A hierarchical gene network was reconstructed using all identified putative genes within a genomic region containing 2.5 cM upstream and downstream of OMT2. Results indicated that numerous important biotic stress‐responsive genes such as RPM1, RGA2, TRIUR3, BURP12 and myosin‐11, were located downstream of melatonin as a master regulator molecule through the OMT2 node. To our knowledge, this is the first report of the identification of an orthologue for Lr19 in Ae. tauschii, which provides insights into the possible regulatory route of this gene.

Identification and Validation of Quantitative Trait Loci Mapping for Spike-Layer Uniformity in Wheat.

Spike-layer uniformity (SLU), the consistency of the spike distribution in the vertical space, is an important trait. It directly affects the yield potential and appearance. Revealing the genetic basis of SLU will provide new insights into wheat improvement. To map the SLU-related quantitative trait loci (QTL), 300 recombinant inbred lines (RILs) that were derived from a cross between H461 and Chinese Spring were used in this study. The RILs and parents were tested in fields from two continuous years from two different pilots. Phenotypic analysis showed that H461 was more consistent in the vertical spatial distribution of the spike layer than in Chinese Spring. Based on inclusive composite interval mapping, four QTL were identified for SLU. There were two major QTL on chromosomes 2BL and 2DL and two minor QTL on chromosomes 1BS and 2BL that were identified. The additive effects of QSlu.sicau-1B, Qslu.sicau-2B-2, and QSlu.sicau-2D were all from the parent, H461. The major QTL, QSlu.sicau-2B-2 and QSlu.sicau-2D, were detected in each of the conducted trials. Based on the best linear unbiased prediction values, the two loci explained 23.97% and 15.98% of the phenotypic variation, respectively. Compared with previous studies, the two major loci were potentially novel and the two minor loci were overlapped. Based on the kompetitive allele-specific PCR (KASP) marker, the genetic effects for QSlu.sicau-2B-2 were validated in an additional RIL population. The genetic effects ranged from 26.65% to 32.56%, with an average value of 30.40%. In addition, QSlu.sicau-2B-2 showed a significant (p < 0.01) and positive influence on the spike length, spikelet number, and thousand kernel weight. The identified QTL and the developed KASP marker will be helpful for fine-mapping these loci, finally contributing to wheat breeding programs in a marker-assisted selection way.

Genetic dissection for head blast resistance in wheat using two mapping populations.

Wheat head blast is a dangerous fungal disease in South America and has recently spread to Bangladesh and Zambia, threatening wheat production in those regions. Host resistance as an economical and environment-friendly management strategy has been heavily relied on, and understanding the resistance loci in the wheat genome is very helpful to resistance breeding. In the current study, two recombinant inbred line (RIL) populations, Alondra/Milan (with 296 RILs) and Caninde#2/Milan-S (with 254 RILs and Milan-S being a susceptible variant of Milan), were used for mapping QTL associated with head blast resistance in field experiments. Phenotyping was conducted in Quirusillas and Okinawa, Bolivia, and in Jashore, Bangladesh, during the 2017–18 and 2018–19 cropping cycles. The DArTseq® technology was employed to genotype the lines, along with four STS markers in the 2NS region. A QTL with consistent major effects was mapped on the 2NS/2AS translocation region in both populations, explaining phenotypic variation from 16.7 to 79.4% across experiments. Additional QTL were detected on chromosomes 2DL, 7AL, and 7DS in the Alondra/Milan population, and 2BS, 4AL, 5AS, 5DL, 7AS, and 7AL in the Caninde#2/Milan-S population, all showing phenotypic effects <10%. The results corroborated the important role of the 2NS/2AS translocation on WB resistance and identified a few novel QTL for possible deployment in wheat breeding. The low phenotypic effects of the non-2NS QTL warrantee further investigation for novel QTL with higher and more stable effects against WB, to alleviate the heavy reliance on 2NS-based resistance.

Identification and Validation of a Novel Major QTL Controlling Leaf Pubescence in the Chinese Wheat Landrace ‘Baimaomai’

Leaf pubescence is an important trait closely associated with plant adaptability to specialized habitats. Baimaomai (BMM) is a wheat (Triticum aestivum L.) landrace originating from the high-altitude, drought-prone environment of Sichuan Province, China with long, dense leaf pubescence. A population of 234 recombinant inbred lines (F10) developed from the cross between Chuanmai104 (CM104), which lacks leaf pubescence, and BMM with pubescent leaves, was used to conduct a phenotypic evaluation of leaf pubescence. Three quantitative trait loci (QTLs) were detected on chromosome arms 7BS, 3DL and 3AL using a high-density wheat 50K single-nucleotide polymorphism array in four environments. The QTLs were designated QLp.saas-7BS, QLp.saas-3DL and QLp.saas-3AL. QLp.saas-3AL, derived from BMM, and QLp.saas-3DL, derived from CM104, were new minor-effect loci. QLp.saas-7BS, derived from BMM, was a novel major-effect locus detected in all environments and was localized in a 0.48 Mb interval on chromosome arm 7BS based on the wheat ‘Chinese Spring’ reference genome. QLp.saas-7BS explained up to 40.77% of the total phenotypic variance. KASP markers tightly linked to QLp.saas-7BS were developed and verified. The present results provide valuable information for further fine mapping, cloning, and marker-assisted selection with QLp.saas-7BS in wheat.

Fine mapping of the tiller inhibition gene TIN4 contributing to ideal plant architecture in common wheat.

A tiller inhibition gene, TIN4, was mapped to an approximately 311kb genomic interval on chromosome arm 2DL of wheat. The tiller is one of the key components of plant morphological architecture and a central agronomic trait affecting spike number in wheat. Low tiller number has been proposed as a major component of crop ideotypes for high yield potential. In this study, we characterized the development of tillering in near-isogenic lines (NIL7A and NIL7B), indicating that the TIN4 gene inhibited the growth of tillering buds and negatively regulated tiller number. Low-tillering was controlled by a single gene (TIN4) located on chromosome 2DL by genetic analysis and bulked segregant RNA-seq analysis. A total of 17 new polymorphic markers were developed in this study, and 61 recombinants were identified in the secondary F2 population containing 4,266 individuals. TIN4 was finally mapped on a 0.35cM interval, co-segregated with molecular marker M380, within a 311kb genomic interval of the wheat cultivar Chinese Spring reference genome sequence that contained twelve predicted genes. Yield experiments showed that the yield of low-tillering lines was higher than that of high-tillering lines at a higher density. Overall, this study provides a foundation for the construction of a low-tillering ideotype for improving wheat yield and further cloning TIN4 by map-based cloning approach.

QTL mapping of drought tolerance at germination stage in wheat using the 50 K SNP array

AbstractDrought is a major concern among abiotic stresses in wheat (Triticum aestivum L.) production. Breeding resistant cultivars are the most effective means to manage drought stress. F6 recombinant inbred lines (RIL) derived from the cross of Berkut/Worrakatta were used to identify quantitative trait loci (QTL) for drought tolerance at germination stage under treatment of PEG6000 using the wheat 50 K single nucleotide polymorphism (SNP) array. Twenty-eight linkage groups were constructed, covering a length of 2220.26 cM. Eighteen QTL were detected based on the drought tolerance coefficients and D-value, explaining 2.7–6.5% of the phenotypic variances, in which 15 were likely to be novel. Three QTL, QGR.xjau-5AS, QCL.xjau-5AS and QD.xjau-5AS for GR, CL and D-value, respectively, at physical positions of 11.70–20.61 Mb between markers AX-111258240 and AX-94458300 on chromosome 5AS accounted for 3.4–4.8% of the phenotypic variances. Three QTL, QGP.xjau-5DL, QSH.xjau-5DL and QD.xjau-5DL for GP, SH and D-value, respectively, were flanked by markers AX-94524442 and AX-110998507 at 560.42–567.39 Mb on chromosome 5DL, accounting for 4.4–6.5% of the phenotypic variances. In addition, the candidate genes TraesCS5A02G022100, TraesCS5B02G014200 and TraesCS5D02G563900 were predicted. Based on transcriptional expression data, the results showed that the expression level of TaGATAs-5A, TaUbox-5B and TaGSTP-5D changed with the increase of treatment time under drought stress in tolerant and sensitive varieties. These are interesting targets in mining drought tolerance genes and the improvement of drought tolerance in wheat.