Chromosome 3BS Research Articles

Dissecting the Genetic Complexity of Fusarium Crown Rot Resistance in Wheat

Fusarium crown rot (FCR) is one of the most important diseases of wheat (Triticum aestivum L.). FCR is mainly caused by the fungal pathogens Fusarium culmorum and F. pseudograminearum. In order to identify new sources of resistance to FCR and to dissect the complexity of FCR resistance, a panel of 161 wheat accessions was phenotyped under growth room (GR) and greenhouse conditions (GH). Analysis of variance showed significant differences in crown rot development among wheat accessions and high heritability of genotype-environment interactions for GR (0.96) and GH (0.91). Mixed linear model analysis revealed seven novel quantitative trait loci (QTLs) linked to F. culmorum on chromosomes 2AL, 3AS, 4BS, 5BS, 5DS, 5DL and 6DS for GR and eight QTLs on chromosomes on 3AS, 3BS, 3DL, 4BS (2), 5BS, 6BS and 6BL for GH. Total phenotypic variances (R²) explained by the QTLs linked to GR and GH were 48% and 59%, respectively. In addition, five favorable epistasis interactions among the QTLs were detected for both GR and GH with and without main effects. Epistatic interaction contributed additional variation up to 21% under GR and 7% under GH indicating strong effects of environment on the expression of QTLs. Our results revealed FCR resistance responses in wheat to be complex and controlled by multiple QTLs.

A spontaneous wheat-Aegilops longissima translocation carrying Pm66 confers resistance to powdery mildew.

A spontaneous Robertsonian T4SlS·4BL translocation chromosome carrying Pm66 for powdery mildew resistance was discovered and confirmed by RNA-seq, molecular marker, and in situ hybridization analyses. Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a severe disease of bread wheat worldwide. Discovery and utilization of resistance genes to powdery mildew from wild relatives of wheat have played important roles in wheat improvement. Aegilops longissima, one of the S-genome diploid wild relatives of wheat, is a valuable source of disease and pest resistance for wheat. Chromosome 4Sl from Ae. longissima confers moderate resistance to powdery mildew. In this study, we conducted RNA-seq on a putative Chinese Spring (CS)-Ae. longissima 4Sl(4B) disomic substitution line (TA3465) to develop 4Sl-specific markers to assist the transfer of a Bgt resistance gene from 4Sl by induced homoeologous recombination. A pairwise comparison of genes between CS and TA3465 demonstrated that a number of genes on chromosome 4BS in CS were not expressed in TA3465. Analysis of 4B- and 4Sl-specific molecular markers showed that 4BS and 4SlL were both missing in TA3465, whereas 4BL and 4SlS were present. Further characterization by genomic and fluorescent in situ hybridization confirmed that TA3465 carried a spontaneous Robertsonian T4SlS·4BL translocation. Powdery mildew tests showed that TA3465 was resistant to 10 of 16 Bgt isolates collected from different regions of China, whereas CS was susceptible to all those Bgt isolates. The powdery mildew resistance gene(s) in TA3465 was further mapped to the short arm of 4Sl and designated as Pm66.

QTL mapping of adult plant resistance to stripe rust and leaf rust in a Fuyu 3/Zhengzhou 5389 wheat population

Stripe or yellow rust (YR) and leaf rust (LR) cause large losses in wheat production worldwide. Resistant cultivars curtail the levels of losses. The present study aimed to identify quantitative trait loci (QTL) for YR and LR resistance in 147 F2:6 recombinant inbred lines (RIL) derived from the cross Fuyu 3/Zhengzhou 5389. The RIL population and parents were genotyped with the Wheat55K single nucleotide polymorphism (SNP) array and simple sequence repeat (SSR) markers. All materials were also phenotyped for YR severity at Mianyang in Sichuan province and Baoding in Hebei province in the 2015/2016, 2016/2017, and 2017/2018 cropping seasons, and LR severity at Zhoukou in Henan province and at Baoding in 2017/2018. Eleven QTL for YR resistance and five for LR resistance were detected using inclusive composite interval mapping (IciMapping). Four of these QTL on chromosomes 1BL, 2BS, 3AL, and 5AL conferred resistance to both YR and LR. The QTL on 1BL was Lr46/Yr29, and that on 7BL might be Lr68. The QTL on chromosome 2BS was detected at a similar position to previously detected loci. QYr.hebau-3AL/QLr.hebau-3AL, QYr.hebau-5AL/QLr.hebau-5AL, QYr.hebau-7DL, QYr.hebau-4BS, QYr.hebau-6DL, and QYr.hebau-2AS are likely to be new. An SSR marker for QYr.hebau-7DL was developed and validated in a diverse wheat panel from China, suggesting effectiveness in different genetic backgrounds. These QTL with closely linked SNP and SSR markers could be useful for marker-assisted selection in wheat breeding programs targeting durable resistance to both diseases.

Anther Extrusion and Its Association with Fusarium Head Blight in CIMMYT Wheat Germplasm

Pronounced anther extrusion (AE) is associated with field resistance to Fusarium head blight (FHB), one of the most devastating diseases of wheat globally. In this study, two recombinant inbred line (RIL) populations were used to map quantitative trait loci (QTL) for AE and field FHB resistance and to investigate the association of both traits at the genetic level. Furthermore, two panels of International Maize and Wheat Improvement Center (CIMMYT) wheat breeding lines were evaluated to describe the phenotypic association between the two traits in detail. Highly significant negative correlation was identified between AE and FHB severity in the two populations and the two panels, with r-values ranging from 0.55 to 0.74. QTL analysis in the two RIL populations identified 12 QTL for AE and nine for FHB resistance, of which five QTL located on chromosomes 3BL, 4BS, 4DS, 5AL, and 5BL were associated with both AE and FHB, collectively explaining over 50% of phenotypic variation for FHB. The QTL on chromosomes 4BS, 4DS, 5AL, and 5BL were closely linked to Rht-B1, Rht-D1, Vrn-A1, and Vrn-B1 genes, respectively. In conclusion, AE is closely related to field FHB resistance and could be used as a morphological marker in wheat breeding for field FHB resistance.

ANÁLISIS DE MARCADORES MOLECULARES PARA EL ÍNDICE DE FERTILIDAD DE ESPIGA Y CARACTERES ASOCIADOS EN UNA POBLACIÓN DE LÍNEAS ENDOCRIADAS RECOMBINANTES DE TRIGO PAN.

Spike fertility index (SF) has been well established as an ecophysiological trait related to grain number per unit area and a promising selection target in wheat breeding programs. Scarce information on the molecular basis of SF is available thus far. In this study, a preliminary molecular marker analysis was carried out in a RIL population derived from the cross between two Argentinean cultivars with contrasting SF to identify candidate genomic regions associated with SF. Twenty-four microsatellites and two functional markers that had been found to co-segregate with SF in a bulked-segregant analysis of the F3 generation of the population were analyzed. Phenotypic data were collected from three field experiments carried out during 2013, 2014 and 2015 growing seasons at Balcarce, Argentina. Two genomic regions associated with SF in chromosomes 5BS and 7AS were detected, which merit further investigation. Key words: selection, genomic regions, grain number, yield, QTL, spike fertility index, fruiting efficiency

QTL mapping for grain yield-related traits in bread wheat via SNP-based selective genotyping.

We identified four chromosome regions harboring QTL for grain yield-related traits, and breeder-friendly KASP markers were developed and validated for marker-assisted selection. Identification of major stable quantitative trait loci (QTL) for grain yield-related traits is important for yield potential improvement in wheat breeding. In the present study, 266 recombinant inbred lines (RILs) derived from a cross between Zhongmai 871 (ZM871) and its sister line Zhongmai 895 (ZM895) were evaluated for thousand grain weight (TGW), grain length (GL), grain width (GW), and grain number per spike (GNS) in 10 environments and for grain filling rate in six environments. Sixty RILs, with 30 higher and 30 lower TGW, respectively, were genotyped using the wheat 660K SNP array for preliminary QTL mapping. Four genetic regions on chromosomes 1AL, 2BS, 3AL, and 5B were identified to have a significant effect on TGW-related traits. A set of Kompetitive Allele Specific PCR markers were converted from the SNP markers on the above target chromosomes and used to genotype all 266 RILs. The mapping results confirmed the QTL named Qgw.caas-1AL, Qgl.caas-3AL, Qtgw.caas-5B, and Qgl.caas-5BS on the targeted chromosomes, explaining 5.0-20.6%, 5.7-15.7%, 5.5-17.3%, and 12.5-20.5% of the phenotypic variation for GW, GL, TGW, and GL, respectively. A novel major QTL for GNS on chromosome 5BS, explaining 5.2-15.2% of the phenotypic variation, was identified across eight environments. These QTL were further validated using BC1F4 populations derived from backcrosses ZM871/ZM895//ZM871 (121 lines) and ZM871/ZM895//ZM895 (175 lines) and 186 advanced breeding lines. Collectively, selective genotyping is a simple, economic, and effective approach for rapid QTL mapping and can be generally applied to genetic mapping studies for important agronomic traits.

Functional mutation allele mining of plant architecture and yield-related agronomic traits and characterization of their effects in wheat

BackgroundWheat mutant resources with phenotypic variation have been developed in recent years. These mutants might carry favorable mutation alleles, which have the potential to be utilized in the breeding process. Plant architecture and yield-related features are important agronomic traits for wheat breeders and mining favorable alleles of these traits will improve wheat characteristics.ResultsHere we used 190 wheat phenotypic mutants as material and by analyzing their SNP variation and phenotypic data, mutation alleles for plant architecture and yield-related traits were identified, and the genetic effects of these alleles were evaluated. In total, 32 mutation alleles, including three pleiotropic alleles, significantly associated with agronomic traits were identified from the 190 wheat mutant lines. The SNPs were distributed on 12 chromosomes and were associated with plant height (PH), tiller number, flag leaf angle (FLA), thousand grain weight (TGW), and other yield-related traits. Further phenotypic analysis of multiple lines carrying the same mutant allele was performed to determine the effect of the allele on the traits of interest. PH-associated SNPs on chromosomes 2BL, 3BS, 3DL, and 5DL might show additive effects, reducing PH by 10.0 cm to 31.3 cm compared with wild type, which means that these alleles may be favorable for wheat improvement. Only unfavorable mutation alleles that reduced TGW and tiller number were identified. A region on chromosome 5DL with mutation alleles for PH and TGW contained several long ncRNAs, and their sequences shared more than 90% identity with cytokinin oxidase/dehydrogenase genes. Some of the mutation alleles we mined were colocalized with previously reported QTLs or genes while others were novel; these novel alleles could also result in phenotypic variation.ConclusionOur results demonstrate that favorable mutation alleles are present in mutant resources, and the region between 409.5 to 419.8 Mb on chromosome 5DL affects wheat plant height and thousand grain weight.

Haplotype block analysis of an Argentinean hexaploid wheat collection and GWAS for yield components and adaptation

BackgroundIncreasing wheat (Triticum aestivum L.) production is required to feed a growing human population. In order to accomplish this task a deeper understanding of the genetic structure of cultivated wheats and the detection of genomic regions significantly associated with the regulation of important agronomic traits are necessary steps. To better understand the genetic basis and relationships of adaptation and yield related traits, we used a collection of 102 Argentinean hexaploid wheat cultivars genotyped with the 35k SNPs array, grown from two to six years in three different locations. Based on SNPs data and gene-related molecular markers, we performed a haplotype block characterization of the germplasm and a genome-wide association study (GWAS).ResultsThe genetic structure of the collection revealed four subpopulations, reflecting the origin of the germplasm used by the main breeding programs in Argentina. The haplotype block characterization showed 1268 blocks of different sizes spread along the genome, including highly conserved regions like the 1BS chromosome arm where the 1BL/1RS wheat/rye translocation is located. Based on GWAS we identified ninety-seven chromosome regions associated with heading date, plant height, thousand grain weight, grain number per spike and fruiting efficiency at harvest (FEh). In particular FEh stands out as a promising trait to raise yield potential in Argentinean wheats; we detected fifteen haplotypes/markers associated with increased FEh values, eleven of which showed significant effects in all three evaluated locations. In the case of adaptation, the Ppd-D1 gene is consolidated as the main determinant of the life cycle of Argentinean wheat cultivars.ConclusionThis work reveals the genetic structure of the Argentinean hexaploid wheat germplasm using a wide set of molecular markers anchored to the Ref Seq v1.0. Additionally GWAS detects chromosomal regions (haplotypes) associated with important yield and adaptation components that will allow improvement of these traits through marker-assisted selection.

Identification of a conserved ph1b-mediated 5DS–5BS crossing over site in soft-kernel durum wheat (Triticum turgidum subsp. durum) lines

Genetic recombination is the major mechanism and basis of genetic diversity and crop improvement. Recombination typically occurs between homologous chromosomes. In wheat, however, it can also occur between homoeologous chromosomes if a functional Ph1 (Pairing homoeologous-1) locus is absent (ph1b). Recently, the genes for soft kernel trait on the distal 5DS chromosome of common wheat were transferred into chromosome 5BS of durum wheat through ph1b-mediated recombination. Several independent 5DS–5BS recombination lines carrying the 5DS translocation were obtained. However, the specific size of the translocation and region where the translocation breakpoint occurred was not determined for any of the lines. In the present study, four independent 5DS–5BS recombination lines were investigated and the translocation breakpoints were fine-mapped and sequenced. All the analyzed lines lost a 5BS fragment of ~ 20,742,425 bp and gained a 5DS fragment of ~ 28,163,252 bp. The recombination breakpoint in each line was located within a predicted gene in a conserved 39 bp region, suggesting the presence of a recombination hotspot. The information obtained was then used to develop a KASP marker diagnostic for the 5DS–5BS translocation. Finally, a subset of the soft durum wheat lines exhibiting varying degrees of kernel hardness was analyzed through genomic in situ hybridization (GISH) and scanning electron microscopy (SEM). From both the GISH and SEM analyses no major differences were detected among the lines, indicating that factors other than ph1b-mediated recombination or endosperm morphology were responsible for the variation in kernel hardness.

Structure and expression analysis of TaGW5 in common wheat

OsGW5 (OsGSE) regulates cell proliferation and is involved in regulating grain size and thousand-grain weight in rice. Our knowledge about its wheat ortholog TaGW5 is limited. In the present study, we characterized the structure and expression of TaGW5 at molecular level in wheat and predicted the cis-elements and transcription factor binding sites (TFBS) of its promoter. The GW5 orthologs in barley (HvGW5), rice (OsGW5), Triticum turgidum L. (TtGW5) and Brachypodium distachyon (BdGW5) were also identified for comparative analyses. TaGW5 was mapped onto the short arms of group 1 chromosomes (1AS, 1BS, and 1DS). Multiple alignments indicated GW5 possesses three exons and two introns in all the analyzed species except for rice and the exon-intron junction composed of exon 2 and intron 2 was highly conserved. GW5 has a conserved domain (DUF 4005) and two neighboring IQ domains and was mainly expressed in wheat young spikes, in barley immature inflorescences and in rice anthers. Drought, heat and biotic-stress treatments had no significant effects on HvGW5 and TaGW5 expression. Significant correlation between the expression patterns of predicted transcription factor ABF2 and OsGW5 was also detected. Taken together, these results broaden our understanding of GW5 in wheat, barley and rice and will be helpful for further manipulating GW5 and uncovering its roles in plants

Grain protein content and thousand kernel weight QTLs identified in a durum × wild emmer wheat mapping population tested in five environments.

Genetic dissection of GPC and TKW in tetraploid durum × WEW RIL population, based on high-density SNP genetic map, revealed 12 GPC QTLs and 11 TKW QTLs, with favorable alleles for 11 and 5 QTLs, respectively, derived from WEW. Wild emmer wheat (Triticum turgidum ssp. dicoccoides, WEW) was shown to exhibit high grain protein content (GPC) and therefore possess a great potential for improvement of cultivated wheat nutritional value. Genetic dissection of thousand kernel weight (TKW) and grain protein content (GPC) was performed using a high-density genetic map constructed based on a recombinant inbred line (RIL) population derived from a cross between T. durum var. Svevo and WEW acc. Y12-3. Genotyping of 208 F6 RILs with a 15K wheat single nucleotide polymorphism (SNP) array yielded 4166 polymorphic SNP markers, of which 1510 were designated as skeleton markers. A total map length of 2169cM was obtained with an average distance of 1.5cM between SNPs. A total of 12 GPC QTLs and 11 TKW QTLs were foundunder five different environments. No significant correlations were found between GPC and TKW across all environments. Four major GPC QTLs with favorable alleles from WEW were found on chromosomes 4BS, 5AS, 6BS and 7BL. The 6BS GPC QTL coincided with the physical position of the NAC transcription factor TtNAM-B1, underlying the cloned QTL, Gpc-B1. Comparisons of the physical intervals of the GPC QTLs described here with the results previously reported in other durum × WEW RIL population led to the discovery of seven novel GPC QTLs. Therefore, our research emphasizes the importance of GPC QTL dissection in diverse WEW accessions as a source of novel alleles for improvement of GPC in cultivated wheat.

QTLs for cell membrane stability and flag leaf area under drought stress in a wheat RIL population

A population of 206 recombinant inbred lines (RILs F9–F10) derived from wheat cross WL711/C306 was phenotyped for morpho-physiological traits such as flag leaf area (FLA), flag leaf length (FLL), flag leaf width (FLW), and cell membrane stability (CMS) under water deficit stress (WDS) environment. High yielding cultivar, WL711 had higher FLA than the medium yielding cultivar C306 across trials under both environments. Parent cultivar C306 maintained membrane integrity while WL711 showed higher membrane damage under WDS. The RIL population showed considerable variation, normal distribution and transgressive segregation for FLA, FLL, FLW and CMS under WDS. The genetic linkage map of WL711/C306 RIL population was constructed comprising of 346 markers. The total map distance was 4526.8 cM with an averaged interval of 12.9 cM between adjacent markers. Major consistent QTL for FLA, FLL, FLW, and CMS were identified on chromosomes 2DS and 3BS respectively in the WL711/C306 RIL population under WDS. The major QTL for FLA, qFLAWD.2D.1 which expressed in multiple environments and for CMS, qCMSWD.3B.3 and qCMSWD.3B.4, accounted for a large proportion of phenotypic variance (PV) with positive allele being contributed by C306, a drought resistant (DR) parent. QTL qFLAWD.2D.1 for FLA co-located with QTL for grain number (GN) and days to flowering (DTF) while QTL qCMSWD.3B.3 and qCMS.3B.4 co-located with QTL for grain yield and its components, days to flowering, canopy temperature and coleoptiles length as reported in our previous publications on the WL711/C306 population (Shukla et al. in Euphytica 203:449–467, 2015; Singh et al. in J Plant Biochem Biotechnol 24:324–330, 2015). Two candidate genes Ghd7 for grain yield and heading date and OsCDK4 for calcium dependent protein kinases were identified in the 2DS and 3BS QTL regions respectively on comparison with gene content of rice chromosomes 7 and 1 respectively. Hence, QTLs qFLAWD.2D.1 and qCMSWD.3B.3 are potential target regions for fine mapping and marker assisted selection for FLA and CMS respectively in wheat under water deficit environments.

QTL Mapping and Transcriptome Analysis to Identify Differentially Expressed Genes Induced by Septoria Tritici Blotch Disease of Wheat

Resistance to Septoria tritici blotch (STB) is an economically important trait in many wheat-breeding programs across the world. Several quantitative trait loci (QTL) for STB resistance were identified in wheat but due to the dynamic pathogen population it is necessary to continuously identify new resistance genes/QTL and determine the underlying resistance mechanism. In this work, we integrated QTL mapping and transcriptome profiling to identify candidate genes underlying QTL associated with STB resistance in bread wheat at the seedling stage. The results revealed four QTL on chromosomes 1BS, 1BL, 3AS and 3DL for STB resistance. Among these, two QTL on 2BL and 3DL were mapped for chlorosis, necrosis and pycnidia while the other two on 1BS and 3AS were associated with necrosis and pycnidia. Among the four identified QTL, genes were identified in three QTL (1BS, 2BL and 3DL). In total, 238 differentially expressed genes (DEGs) were localized in 1BS, 16 DEGs in 2BL and 80 DEGs in 3DL QTL region respectively. F-box protein, NBS-LRR disease resistance genes and receptor-like protein kinase were the most over-represented. The results emphasize the importance of integrating QTL and transcriptome analysis to accelerate the identification of key genes underlying the traits of interest.

A 4-bp deletion in the 5\u2019UTR of TaAFP-B is associated with seed dormancy in common wheat (Triticum aestivum L.)

BackgroundAFP is a negative regulator of ABA signaling that promotes ABI5 protein degradation and weakens regulation of ABA signaling by targeting upstream genes of ABI5, and TaABI5 gene was seed-specific, and accumulated during wheat grain maturation and dormancy acquisition, which played an important role in seed dormancy; TaAFP has a conserved domain with AFP, so TaAFP may also play an important role in seed dormancy in wheat.ResultsTwo allelic variants of TaAFP were identified on chromosome 2BS in common wheat, and designated as TaAFP-B1a and TaAFP-B1b. Sequence analysis showed a 4-bp deletion in the 5’UTR region of TaAFP-B1b compared with TaAFP-B1a. Based on the 4-bp deletion, a co-dominant functional marker of TaAFP-B was developed and designated as AFPB. The genotype generating a 203-bp fragment (TaAFP-B1b) was more resistant to pre-harvest sprouting than the genotype producing a 207-bp fragment (TaAFP-B1a) in a test of 91 white-grained Chinese wheat cultivars and advanced lines. The average germination index(GI) values of TaAFP-B1a and that of TaAFP-B1b were 45.18 and 30.72%, respectively, indicating a significant difference (P < 0.001). Moreover, the 4-bp deletion located in the 5’UTR not only affected the transcription level of TaAFP-B but also affected the mRNA decay, reduced the translation level of GUS and tdTomatoER and GUS activity in wheat leaves of transient expression. The transcript expression and the mRNA half-life value of TaAFP-B1a in developing seeds and mature seeds were much higher than those of TaAFP-B1b.ConclusionWe identified a 4-bp InDel in the 5’UTR of TaAFP-B, which affected the mRNA transcription level, mRNA decay, translation levels of GUS and tdTomatoER, GUS activity, and was significantly associated with seed dormancy in common wheat. A functional marker was developed and validated based on this InDel.

Meta-analysis of QTL for Fusarium head blight resistance in Chinese wheat landraces

Epidemics of Fusarium head blight (FHB), incited by Fusarium graminearum Schwabe, in wheat cause significant reductions in grain yield and quality. Numerous quantitative trait loci (QTL) for FHB resistance have been reported from Chinese sources. However, the relationships among QTL from different landraces have not been characterized. We earlier mapped QTL for FHB resistance using low-density maps developed from five recombinant inbred line (RIL) populations involving Chinese landraces ‘Haiyanzhong’ (HYZ), ‘Wangshuibai’ (WSB), ‘Baishanyuehuang’ (BSYH), ‘Huangfangzhu’ (HFZ), and ‘Huangcandou’ (HCD) as FHB resistant parents. In this study, we used maps of single nucleotide polymorphisms (SNP) developed from the five populations and identified 31 QTL on 16 chromosomes; 10 QTL were new. We constructed a consensus map and identified six meta-QTL (MQTL) and SNP within the MQTL regions using meta-analysis. Two of the MQTL were on chromosome 3BS (3BSd and 3BSc), and one on each of chromosomes 3A, 2D, 3D, and 4D. Twenty-two SNP closely linked to MQTL were converted into breeder friendly Kompetitive Allele Specific Polymerase Chain Reaction (KASP) assays, which should be useful for marker-assisted selection in breeding programs.

Genome-wide association study of pre-harvest sprouting tolerance using a 90K SNP array in common wheat (Triticum aestivum L.).

Three major loci for pre-harvest sprouting tolerance (PHST) were mapped on chromosomes 1AL, 3BS, and 6BL, and two CAPS and one dCAPS markers were validated. Sixteen lines with favorable alleles and increased PHST were identified. Pre-harvest sprouting (PHS) significantly affects wheat grain yield and quality. In the present study, the PHS tolerance (PHST) of 192 wheat varieties (lines) was evaluated by assessment of field sprouting, seed germination index, and period of dormancy in different environments. A high-density Illumina iSelect 90K SNP array was used to genotype the panel. A genome-wide association study (GWAS) based on single- and multi-locus mixed linear models was used to detect loci for PHST. The single-locus model identified 23 loci for PHST (P < 0.0001) and explained 6.0-18.9% of the phenotypic variance. Twenty loci were consistent with known quantitative trait loci (QTLs). Three single-nucleotide polymorphism markers closely linked with three major loci (Qphs.ahau-1A, Qphs.ahau-3B, and Qphs.ahau-6B) on chromosomes 1AL, 3BS, and 6BL, respectively, were converted to two cleaved amplified polymorphic sequences (CAPS) and one derived-CAPS markers, and validated in 374 wheat varieties (lines). The CAPS marker EX06323 for Qphs.ahau-6B co-segregated with a novel major QTL underlying PHST in a recombinant inbred line population raised from the cross Jing 411 × Wanxianbaimaizi. Linear regression showed a clear dependence of PHST on the number of favorable alleles. Sixteen varieties showing an elevated degree of PHST were identified and harbored more than 16 favorable alleles. The multi-locus model detected 39 marker-trait associations for PHST (P < 0.0001), of which five may be novel. Six loci common to the two models were identified. The combination of the two GWAS methods contributes to efficient dissection of the complex genetic mechanism of PHST.

Molecular characterization of bacterial leaf streak resistance in hard winter wheat.

Bacterial leaf streak (BLS) caused by Xanthomonas campestris pv. translucens is one of the major bacterial diseases threatening wheat production in the United States Northern Great Plains (NGP) region. It is a sporadic but widespread wheat disease that can cause significant loss in grain yield and quality. Identification and characterization of genomic regions in wheat that confer resistance to BLS will help track resistance genes/QTLs in future wheat breeding. In this study, we evaluated a hard winter wheat association mapping panel (HWWAMP) containing 299 hard winter wheat lines from the US hard winter wheat growing region for their reactions to BLS. We observed a range of BLS responses among the lines, importantly, we identified ten genotypes that showed a resistant reaction both in greenhouse and field evaluation. ­Genome-wide association analysis with 15,990 SNPs was conducted using an exponentially compressed mixed linear model. Five genomic regions (p < 0.001) that regulate the resistance to BLS were identified on chromosomes 1AL, 1BS, 3AL, 4AL, and 7AS. The QTLs Q.bls.sdsu-1AL, Q.bls.sdsu-1BS, Q.bls.sdsu-3AL, Q.bls.sdsu-4AL, and Q.bls.sdsu-7AS explain a total of 42% of the variation. In silico analysis of sequences in the candidate regions on chromosomes 1AL, 1BS, 3AL, 4AL, and 7AS identified 10, 25, 22, eight, and nine genes, respectively with known plant defense-related functions. Comparative analysis with rice showed two syntenic regions in rice that harbor genes for bacterial leaf streak resistance. The ten BLS resistant genotypes and SNP markers linked to the QTLs identified in our study could facilitate breeding for BLS resistance in winter wheat.

Mutation of a histidine-rich calcium-binding-protein gene in wheat confers resistance to Fusarium head blight.

Head or ear blight, mainly caused by Fusarium species, can devastate almost all staple cereal crops (particularly wheat), resulting in great economic loss and imposing health threats on both human beings and livestock1-3. However, achievement in breeding for highly resistant cultivars is still not satisfactory. Here, we isolated the major-effect wheat quantitative trait locus, Qfhs.njau-3B, which confers head blight resistance, and showed that it is the same as the previously designated Fhb1. Fhb1 results from a rare deletion involving the 3' exon of the histidine-rich calcium-binding-protein gene on chromosome 3BS. Both wheat and Arabidopsis transformed with the Fhb1 sequence showed enhanced resistance to Fusarium graminearum spread. The translation products of this gene's homologs among plants are well conserved and might be essential for plant growth and development. Fhb1 could be useful not only for curbing Fusarium head blight in grain crops but also for improving other plants vulnerable to Fusarium species.

Genome-Wide Association Study Reveals Novel Genomic Regions Associated With High Grain Protein Content in Wheat Lines Derived From Wild Emmer Wheat.

Grain protein content (GPC) and yield are of two important traits in wheat, but their negative correlation has hampered their simultaneous improvement in conventional breeding. Wild emmer wheat (Triticum turgidum ssp. dicoccoides) is an important genetic resource for wheat quality improvement. In this study, we report a genome-wide association study (GWAS) using 13116 DArT-seq markers to characterize GPC in 161 wheat lines derived from wild emmer. Using a general linear model, we identified 141 markers that were significantly associated with GPC, and grouped into 48 QTL regions. Using both general linear model and mixed linear model, we identified four significant markers that were grouped into two novel QTL regions on chromosomes 2BS (QGpc.cd1-2B.1) and 7BL (QGpc.cd1-7B.2). The two QTLs have no negative effects on thousand kernel weight (TKW) and should be useful for simultaneous improvement of GPC and TKW in wheat breeding. Searches of public databases revealed 61 putative candidate/flanking genes related to GPC. The putative proteins of interest were grouped in four main categories: enzymes, kinase proteins, metal transport-related proteins, and disease resistance proteins. The linked markers and associated candidate genes provide essential information for cloning genes related to high GPC and performing marker-assisted breeding in wheat.

Genetic analysis of threshability and other spike traits in the evolution of cultivated emmer to fully domesticated durum wheat.

Genetic mutations in genes governing wheat threshability were critical for domestication. Knowing when these genes mutated during wheat evolution will provide more insight into the domestication process and lead to further exploitation of primitive alleles for wheat improvement. We evaluated a population of recombinant inbred lines derived from a cross between the durum variety Rusty and the cultivated emmer accession PI 193883 for threshability, rachis fragility, and other spike-related traits. Quantitative trait loci (QTL) associated with spike length, spikelets per spike, and spike compactness were primarily associated with known genes such as the pleiotropic domestication gene Q. Interestingly, rachis fragility was not associated with the Q locus, suggesting that this trait, usually a pleiotropic effect of the q allele, can be influenced by the genetic background. Threshability QTL were identified on chromosome arms 2AS, 2BS, and 5AL corresponding to the tenacious glume genes Tg2A and Tg2B as well as the Q gene, respectively, further demonstrating that cultivated emmer harbors the primitive non-free-threshing alleles at all three loci. Genetic analysis indicated that the effects of the three genes are mostly additive, with Q having the most profound effects on threshability, and that free-threshing alleles are necessary at all three loci to attain a completely free-threshing phenotype. These findings provide further insight into the timeline and possible pathways of wheat domestication and evolution that led to the formation of modern day domesticated wheats.