Search

As a devastating wheat disease, Fusarium head blight (FHB) causes large losses in yield and quality. The purpose of this study was to detect the genetic loci for FHB resistance based on a genome-wide association study (GWAS) using two panels genotyped by the Wheat 660 K SNP array and Wheat 90 K SNP array. Phenotypic investigation results showed that 43.6–51.6% of the test varieties showed an infection type of more than 3, indicating that substantial improvement in the level of resistance was urgently required. GWAS detected 1264 SNPs associated with FHB response distributed on all wheat chromosomes. Fourteen QTL were stably detected in two environments. gFHB-1B (582.9~607.9 Mb on chromosome 1B) and gFHB-5A (577.9~596.5 Mb on 5A) were two novel resistance loci that were detected in two wheat panels. Haplotype analysis of both gFHB-1B and gFHB-5A indicated that varieties with gFHB-1B_R and gFHB-5A_R had consistently less FHB than varieties carrying a one or no resistance QTL. The application frequency of the FHB-resistant alleles gFHB-1B and gFHB-5A showed a gradually decreasing trend in FHB resistance breeding. This study identified new genetic loci for the control of FHB and provides useful information for marker-assisted selection in wheat FHB resistance.

Leaf rust, caused by Puccinia triticinaEriksson (Pt), is one of the most severe foliar diseases of wheat. Breeding for leaf rust resistance is a practical and sustainable method to control this devastating disease. Here, we report the identification of Lr47, a broad-spectrum leaf rust resistance gene introgressed into wheat fromAegilops speltoides. The Lr47 gene encodes a coiled-coil nucleotide-binding leucine-rich repeat protein that is both necessary and sufficient to confer Pt resistance, as demonstrated by loss-of-function mutations and transgenic complementation. New Lr47 introgression lines with no or reduced linkage dragwere generated using the ph1b mutation, and a diagnostic molecular marker for Lr47 was developed. The CC domain of the Lr47 protein was not able to induce cell death, nor did it have self-protein interaction. The cloning of Lr47 expands the number of Pt-resistance genes that can be incorporated into multigene transgenic cassettes to control this devastating disease.

Dissecting the genetic basis of seed traits in wheat is impeded by limited genetic polymorphisms and significant variations caused by environmental conditions and seed position in a spikelet. Seed performance is largely determined by endosperm development controlled by spatiotemporal variation in gene activities, which is greatly affected by chromatin status. Here, we mapped genome-wide dynamic distributions of H3K27me3, H3K4me3 and H3K9ac modifications and profiled gene transcription across wheat endosperm development. The combinatorial effects of active and repressive marks ensure spatiotemporal dynamic gene expression, especially for starch biosynthesis. By scanning the transcription factor binding motifs in the H3K9ac peaks, hub regulators were identified from the regulatory network. Strikingly, significant correlations were detected between sequence polymorphisms of hub regulators and variation in seed traits in a germplasm population. Thus, the analysis of genomic regulatory activities together with genetic variation provides a robust approach to dissect seed traits in bread wheat.

Leaf rust (LR) and stripe rust (YR) are important diseases in wheat producing areas worldwide and cause severe yield losses under favorable environmental conditions when susceptible varieties are grown. We determined the genetic basis of resistance to LR and YR in variety Borlaug 100 by developing and phenotyping a population of 198 F6 recombinant inbred lines derived from a cross with the susceptible parent Apav#1. LR and YR phenotyping were conducted for 4 and 3 seasons, respectively, at CIMMYT research stations in Mexico under artificial epidemics. Mendelian segregation analyses indicated that 3–5 LR and 2 YR genes conferred resistance in Borlaug 100. Lr46/Yr29 (1BL), Yr17 (2AS) and Yr30 (3BS) were present in the resistant parent and segregated in the RIL population based on characterization by molecular markers linked to these genes. When present alone, Lr46/Yr29 caused average 13% and 16% reductions in LR and YR severities, respectively, in RILs. Similarly, Yr17 and Yr30 reduced YR severities by 57% and 11%, respectively. The Yr30 and the Yr17 translocation were also associated with 27% and 14% reductions, respectively, in LR severity, indicating that the 3BS and 2AS chromosomal regions likely carry new slow rusting LR resistance genes, temporarily designated as LrB1 and LrB2, respectively. Additive effects of Yr30*Yr17, Yr29*Yr17 and Yr29*Yr30 on YR and LR were significant and reduced YR severities by 56%, 55%, and 45%, respectively, and LR severities by 34%, 40%, and 45%, respectively. Furthermore, interaction between the three genes was also significant, with mean reductions of 70% for YR and 54% for LR severities. Borlaug 100, or any one of the 21 lines with variable agronomic traits but carrying all three co-located resistance loci, can be used as resistance sources in wheat breeding programs.

Breeding resistant wheat variety is the most economic and efficient way to manage wheat stripe rust, while identification of new stripe rust resistance gene and development the molecular markers will be very useful for wheat breeder in the breeding program. In the present study, a recombinant inbred line (RIL) population derived from a cross of the resistant parent KFA/2*KACHU and susceptible parent Apav#1 was used to map stripe rust resistance loci. In combination of GBS genotyping platform and BSR-Seq method, we mapped a new race-specific stripe rust resistance gene on wheat chromosome 5BL at the seedling stage, temporarily named as YrK. TraesCS5B02G330700 encoded a receptor-like kinase was considered as the key candidate gene of YrK based on virus induced gene silencing (VIGS) method. Its expression showed a significant up-regulated at 24h after inoculation and the functional molecular markers were developed based on the polymorphic SNPs in the CDS region. In addition, a total of four adult plant resistance (APR) loci were identified on wheat chromosome 1BL, 2AS, 2BS and 4AL, respectively. Among these, QYr.hazu-1BL and QYr.hazu-2AS was verified as the known resistance genes Lr46/Yr29/Pm39 and Yr17, respectively. QYr.hazu-2BS was a new APR locus and the closely linked SNP markers were converted into breeder-friendly KASP markers.

Melatonin (Mel) is a multifunctional biomolecule found in both animals and plants. In plants, the biosynthesis of Mel from tryptophan (Trp) has been delineated to comprise of four consecutive reactions. However, while the genes encoding these enzymes in rice are well characterized no systematic evaluation of the overall pathway has, as yet, been published for wheat. In the current study, the relative contents of six Mel-pathway-intermediates including Trp, tryptamine (Trm), serotonin (Ser), 5-methoxy tryptamine (5M-Trm), N-acetyl serotonin (NAS) and Mel, were determined in 24 independent tissues spanning the lifetime of wheat. These studies indicated that Trp was the most abundant among the six metabolites, followed by Trm and Ser. Next, the candidate genes expressing key enzymes involved in the Mel pathway were explored by means of metabolite-based genome-wide association study (mGWAS), wherein two TDC genes, a T5H gene and one SNAT gene were identified as being important for the accumulation of Mel pathway metabolites. Moreover, a 463-bp insertion within the T5H gene was discovered that may be responsible for variation in Ser content. Finally, a ASMT gene was found via sequence alignment against its rice homolog. Validations of these candidate genes were performed by in vitro enzymatic reactions using proteins purified following recombinant expression in Escherichia coli, transient gene expression in tobacco, and transgenic approaches in wheat. Our results thus provide the first comprehensive investigation into the Mel pathway metabolites, and a swift candidate gene identification via forward-genetics strategies, in common wheat.

Agrobacterium–mediated transformation is a cost-effective and convenient way to introduce foreign genetic elements to plants. However, only limited protocols successfully generates transgenic plants with fielder, a spring wheat variety. Wheat transformation method with higher efficiency and without genotype restriction is heavily demanded. Here, a heat-shock protocol, independent of Japan Tobacco had been established. Transgenic plants can be obtained from immature embryo within only 60 days by this protocol. Morphogenic regulators Baby boom and Wuschel (BBM-WUS) was proved to promote transformation efficiency for five to six times in wheat when co-infiltrated with agrobacterium containing target construct. Notably, half of the transformants are BBM-WUS free and moreover, the BBM-WUS containing plants could be picked by florescent marker that was co-expressed with BBM-WUS. In conclusion, we managed to establish a new wheat transformation protocol with shorter duration than published protocol.

The durum wheat lines Heller#1 and Dunkler from the International Maize and Wheat Improvement Center Global Wheat Program showed moderate and stable adult plant resistance to leaf rust under high disease pressure over field environments in northwestern Mexico. Leaf rust phenotyping was performed on two recombinant inbred line (RIL) populations derived from crosses of Heller#1 and Dunkler with the susceptible parent Atred#2, conducted under artificially induced Puccinia triticina epidemics in 2013, 2014, 2015, and 2016. The Atred#2 × Heller#1 and Atred#2 × Dunkler populations were genotyped by single nucleotide polymorphism (SNP) platforms and diversity arrays technology markers, respectively. Four leaf rust resistance quantitative trait loci were detected simultaneously in the two RIL populations: Lr46, QLr.cim-2BC, QLr.cim-5BL, and QLr.cim-6BL based on phenotypic data across all four crop seasons. They explained 11.7 to 46.8%, 7.2 to 26.1%, 8.4 to 24.1%, and 12.4 to 28.5%, respectively, of the phenotypic variation for leaf rust resistance in Atred#2 × Heller#1 and 16.3 to 56.6%, 6.7 to 15.7%, 4.1 to 10.1%, and 5.1 to 20.2% of the variation in the Atred#2 × Dunkler population. Only the resistance allele of QLr.cim-2BC was from the susceptible parent Atred#2, and resistance alleles at other loci came from the resistant parents Heller#1 and Dunkler. The SNP markers closely linked to Lr46 and QLr.cim-2BC were converted to kompetitive allele specific PCR markers for use in marker-assisted selection to improve leaf rust resistance through crosses with Heller#1 and Dunkler sources.

Wheat (Triticum aestivum L.) is one of the main food crops in the world and a primary source of zinc (Zn) and iron (Fe) in the human body. The genetic mechanisms underlying related traits have been clarified, thereby providing a molecular theoretical foundation for the development of germplasm resources. In this study, 23,536 high-quality DArT markers were used to map quantitative trait loci (QTL) of grain Zn (GZn) and grain Fe (GFe) concentrations in recombinant inbred lines from Avocet/Chilero. A total of 17 QTLs located on chromosomes 1BL, 2BL, 3BL, 4AL, 4BS, 5AL, 5DL, 6AS, 6BS, 6DS, and 7AS accounted for 0.38–16.62% of the phenotypic variance. QGZn.haust-4AL, QGZn.haust-7AS.1, and QGFe.haust-6BS were detected on chromosomes 4AL, 6BS, and 7AS, accounting for 10.63–16.62% of the phenotypic variance. Four stable QTLs, QGZn.haust-4AL, QGFe.haust-1BL, QGFe.haust-4AL, and QGFe.haust-5DL were located on chromosomes 1BL, 4AL, and 5DL. Three pleiotropic effects locus for GZn and GFe concentrations were located on chromosomes 1BL, 4AL, and 5DL. Two high-throughput Kompetitive Allele Specific PCR markers were developed by closely linking single nucleotide polymorphisms on chromosomes 4AL and 5DL, which were validated by a germplasm panel. Therefore, it is the most important that quantitative trait loci and KASP marker for grain zinc and iron concentrations were developed for utilizing in marker-assisted breeding and biofortification of wheat grain in breeding programs.

Stripe rust is a major disease constraint of wheat production worldwide. Resistance to stripe rust was analyzed using 131 F6 recombinant inbred lines (RILs) derived from a cross between synthetic derived wheat line Soru#1 and wheat cultivar Naxos. The phenotype was evaluated in Mexico and Norway at both seedling and adult plant stages. Linkage groups were constructed based on 90K single-nucleotide polymorphism (SNP), sequence-tagged site, and simple sequence repeat markers. Two major resistance loci conferred by Soru#1 were detected and located on chromosomes 1BL and 4DS. The 1BL quantitative trait loci explained 15.8 to 40.2 and 51.1% of the phenotypic variation at adult plant and seedling stages, respectively. This locus was identified as Yr24/Yr26 based on the flanking markers and infection types. Locus 4DS was flanked by molecular markers D_GB5Y7FA02JMPQ0_238 and BS00108770_51. It explained 8.4 to 27.8 and 5.5% of stripe rust variation at the adult plant and seedling stages, respectively. The 4DS locus may correspond to known resistance gene Yr28 based on the resistance source. All RILs that combine Yr24/Yr26 and Yr28 showed significantly reduced stripe rust severity in all four environments compared with the lines with only one of the genes. SNP marker BS00108770_51 was converted into a breeder-friendly kompetitive allele-specific polymerase chain reaction marker that will be useful to accelerate Yr28 deployment in wheat breeding programs.

The characterization of leaf rust (caused by Puccinia triticina) and stripe rust (caused by Puccinia striiformis f. sp. tritici) resistance genes is the basis for breeding resistant wheat varieties and managing epidemics of these diseases in wheat. A cross between the susceptible wheat variety ‘Apav#1’ and resistant variety ‘Neimai 836’ was used to develop a mapping population containing 148 F5 recombinant inbred lines (RILs). Leaf rust phenotyping was done in field trials at Ciudad Obregón, Mexico, in 2017 and 2018, and stripe rust data were generated at Toluca, Mexico, in 2017 and in Mianyang, Ezhou, and Gansu, China, in 2019. Inclusive complete interval mapping (ICIM) was used to create a genetic map and identify significant resistance quantitative trait loci (QTL) with 2,350 polymorphic markers from a 15K wheat single-nucleotide polymorphism (SNP) array and simple-sequence repeats (SSRs). The pleiotropic multipathogen resistance gene Lr46/Yr29 and four QTL were identified, including two new loci, QLr.hzau-3BL and QYr.hzau-5AL, which explained 3 to 16% of the phenotypic variation in resistance to leaf rust and 7 to 14% of that to stripe rust. The flanking SNP markers for the two loci were converted to Kompetitive Allele-Specific PCR (KASP) markers and used to genotype a collection of 153 wheat lines, indicating the Chinese origin of the loci. Our results suggest that Neimai 836, which has been used as a parent for many wheat varieties in China, could be a useful source of high-level resistance to both leaf rust and stripe rust.

Genetic dissection kernel weight-related traits is of great significance for improving wheat yield potential. As one of the three major yield components of wheat, thousand kernel weight (TKW) was mainly affected by grain length (GL) and grain width (GW). To uncover the key loci for these traits, we carried out a quantitative trait loci (QTL) analysis of an F6 recombinant inbred lines (RILs) population derived from a cross of Henong 5290 (small grain) and 06Dn23 (big grain) with a 50 K single nucleotide polymorphism (SNP) array. A total of 17 stable and big effect QTL, including 5 for TKW, 8 for GL and 4 for GW, were detected on the chromosomes 1B, 2A, 2B, 2D, 4B, 5A, 6A and 6D, respectively. Among these, there were two co-located loci for three traits that were mapped on the chromosome 4BS and 6AL. The QTL on 6AL was the most stable locus and explained 15.4–24.8%, 4.1–8.8% and 15.7–24.4% of TKW, GW and GL variance, respectively. In addition, two more major QTL of GL were located on chromosome arm 2BL and 2DL, accounting for 9.7–17.8% and 13.6–19.8% of phenotypic variance, respectively. In this study, we found one novel co-located QTL associated with GL and TKW in 2DL, QGl.haaf-2DL.2/QTkw.haaf-2DL.2, which could explain 13.6–19.8% and 9.8–10.7% phenotypic variance, respectively. Genetic regions and linked markers of these stable QTL will help to further refine mapping of the corresponding loci and marker-assisted selection (MAS) breeding for wheat grain yield potential improvement.

Fructans (fructo-oligosaccharides) is one of the most important carbohydrates in wheat vegetative tissues and plays a vital role in wheat grain development. One hundred and forty 'Avocet'× 'Chilero' wheat (Triticum aestivum L.) recombinant inbred lines (RILs) were phenotyped for grain fructans at Luoyang and Langzhou field sites in the 2015-2016 and 2016-2017 growing seasons and 2090 Diversity Arrays Technology (DArT) molecular marker data were used to determine genomic regions controlling fructan content. Eleven significant quantitative trait loci (QTL) for wheat fructan content were identified on chromosomes 1B, 2A, 2B, 2D, 3D, 4B, 6B, 6D, 7A, and 7B using the inclusive composite interval mapping (ICIM) method, explaining 4.08% to 27.40% of the phenotypic variance, with seven exceeding 10%. The single environment analysis identified QF.haust-6D.1 and QF.haust-7B in Luoyang, QF.haust-1B, QF.haust-2D, QF.haust-3D, and QF.haust-7B in Lanzhou, and QF.haust-1B, QF.haust-7A, and QF.haust-7B in average phenotypic performance. The multi-environment analysis identified QF.haust-2B and QF.haust-7B, which explained 21.01% and 17.39% of the phenotypic variance, respectively. The positive allele of QF.haust-2B and QF.haust-7B were all originated from Chilero. QF.haust-2B was flanked by the 3957031 and SNP1203021 markers; candidate genes for QF.haust-2B were identified by anchoring marker sequences in the IGWSC wheat genome sequence, involving a variety of biological processes, such as the regulation of ribosomes metabolism in wheat. We also constructed the physical map for wheat grain fructan QF.haust-2B and QF.haust-7B, QF.haust-7B with a physical length of 11.47 Mb between marker SNP3026392 and 3954877 and 103 genes within the segment, in which ten were related to glucose metabolism. This study provides a theoretical basis and technical support for wheat grain fructan content-related genes and molecular marker-assisted breeding.

Since 1984, the ‘Chilero’ spring wheat line developed by CIMMYT has proven to be highly resistant to leaf rust and stripe rust. Amid efforts to understand the basis of resistance of this line, a recombinant inbred line (RIL) population derived from a cross between Avocet and Chilero was studied. The parents and RILs were characterized in field trials for leaf rust and stripe rust in three locations in Mexico between 2012 and 2015 and genotyped with DArT-array, DArT-GBS, and SSR markers. A total of 6,168 polymorphic markers were used to construct genetic linkage maps. Inclusive composite interval mapping detected four colocated resistance loci to both rust diseases and two stripe rust resistant loci in the Avocet × Chilero population. Among these, the quantitative trait locus (QTL) on chromosome 1BL was identified as a pleotropic adult plant resistance gene Lr46/Yr29, whereas QLr.cim-5DS/QYr.cim-5DS was a newly discovered colocated resistance locus to both rust diseases in Chilero. Additionally, one new stripe rust resistance locus on chromosome 7BL was mapped in the current population. Avocet also contributed two minor colocated resistance QTLs situated on chromosomes 1DL and 4BS. The flanking SNP markers can be converted to breeder friendly Kompetitive Allele Specific PCR (KASP) markers for wheat breeding programs.

Leaf rust (LR) and stripe rust (YR) are important diseases of wheat worldwide. We used 148 recombinant inbred lines (RIL) from the cross of Avocet × Kundan for determining and mapping the genetic basis of adult plant resistance (APR) loci. The population was phenotyped LR and YR for three seasons in field trials conducted in Mexico and genotyped with the diversity arrays technology sequencing (DArT-Seq) and simple sequence repeat markers. The final genetic map was constructed using 2,937 polymorphic markers with an average distance of 1.29 centimorgans between markers. Inclusive composite interval mapping identified two co-located APR quantitative trait loci (QTL) for LR and YR, two LR QTL, and three YR QTL. The co-located resistance QTL on chromosome 1BL corresponded to the pleiotropic APR gene Lr46/Yr29. QLr.cim-2BL, QYr.cim-2AL, and QYr.cim-5AS could be identified as new resistance loci in this population. Lr46/Yr29 contributed 49.5 to 65.1 and 49.2 to 66.1% of LR and YR variations, respectively. The additive interaction between detected QTL showed that LR severities for RIL combining four QTL ranged between 5.3 and 25.8%, whereas the lowest YR severities were for RIL carrying QTL on chromosomes 1BL + 2AL + 6AL. The high-density DArT-Seq markers across chromosomes can be used in fine mapping of the targeted loci and development SNP markers.

New leaf rust adult plant resistance (APR) QTL QLr.cim - 6BL was mapped and confirmed the known pleotropic APR gene Lr46 effect on leaf rust in durum wheat line Bairds. CIMMYT-derived durum wheat line Bairds displays an adequate level of adult plant resistance (APR) to leaf rust in Mexican field environments. A recombinant inbred line (RIL) population developed from a cross of Bairds with susceptible parent Atred#1 was phenotyped for leaf rust response at Ciudad Obregon, Mexico, during 2013, 2014, 2015 and 2016 under artificially created epidemics of Puccinia triticina (Pt) race BBG/BP. The RIL population and its parents were genotyped with the 50 K diversity arrays technology (DArT) sequence system and simple sequence repeat (SSR) markers. A genetic map comprising 1150 markers was used to map the resistance loci. Four significant quantitative trait loci (QTLs) were detected on chromosomes 1BL, 2BC (centromere region), 5BL and 6BL. These QTLs, named Lr46, QLr.cim-2BC, QLr.cim-5BL and QLr.cim-6BL, respectively, explained 13.5–60.8%, 9.0–14.3%, 2.8–13.9%, and 11.6–29.4%, respectively, of leaf rust severity variation by the inclusive composite interval mapping method. All of these resistance loci were contributed by the resistant parent Bairds, except for QLr.cim-2BC, which came from susceptible parent Atred#1. Among these, the QTL on chromosome 1BL was the known pleiotropic APR gene Lr46, whereas QLr.cim-6BL, a consistently detected locus, should be a new leaf rust resistance locus in durum wheat. The mean leaf rust severity of RILs carrying all four QTLs ranged from 8.0 to 17.5%, whereas it ranged from 10.9 to 38.5% for three QTLs (Lr46 + 5BL + 6BL) derived from the resistant parent Bairds. Two RILs with four QTLs combinations can be used as sources of complex APR in durum wheat breeding.

Mining of new genetic resources is of paramount importance to combat the alarming spread of stripe rust disease and breakdown of major resistance genes in wheat. We conducted a genome wide association study on 352 un-utilized Afghan wheat landraces against stripe rust resistance in eight locations. High level of disease variation was observed among locations and a core-set of germplasm showed consistence performance. Linkage disequilibrium (LD) decayed rapidly (R2≈0.16 at 0cM) due to germplasm peerless diversity. The mixed linear model resulted in ten marker-trait associations (MTAs) across all environments representing five QTL. The extensively short LD blocks required us to repeat the analysis with less diverse subset of 220 landraces in which R2 decayed below 0.2 at 0.3cM. The subset GWAS resulted in 36 MTAs clustered in nine QTL. The subset analysis validated three QTL previously detected in the full list analysis. Overall, the study revealed that stripe rust epidemics in the geographical origin of this germplasm through time have permitted for selecting novel resistance loci.

The International Maize and Wheat Improvement Center (CIMMYT)-derived durum wheat line Quetru is moderately resistant to leaf rust and displays disease severities of 15–30% at the adult plant stage. Atil C2000 is moderately susceptible to leaf rust at adult plant stage and displays severities of 20–60%. These two durum wheats were crossed with the susceptible line Atred#1, and the respective recombinant inbred line populations were phenotyped for resistance to leaf rust in field trials in four environments in Mexico. The phenotypic segregation in the Atred#1 × Atil C2000 population was attributed to the pleiotropic adult plant resistance (APR) gene Lr46 based on the closest marker cslv46G22 located within 1.4 cM. Lr46 reduced leaf rust severity by 19.6–35.5%, and it was also identified in the Atred#1 × Quetru population, where it reduced leaf rust severity by 28.5–57.7% depending on the environment. We also identified a putatively novel, race-specific APR gene LrQ in Quetru that is located on the Chromosome 6BL and flanked by SNP markers AX-95144243 and AX-95155193. These two markers have been converted into kompetitive allele–specific PCR (KASP) markers and validated in the population. LrQ was ineffective at the seedling stage but reduced leaf rust severity by 37.3% on average over four field trials. The combination of LrQ and Lr46 resulted in significantly reduced leaf rust severity in the field and could be used to breed improved durum wheat varieties.

Leaf rust (LR) is a destructive foliar disease of common wheat (Triticum aestivum L.) worldwide. Growing resistant cultivars with adult plant resistance (APR) are the ideal way to control the disease. The aim of the present study was to map quantitative trait loci (QTL) for leaf rust resistance using 137 doubled-haploid (DH) lines derived from the Pingyuan 50 × Mingxian 169 cross. The same population was previously tested for stripe rust (YR) and powdery mildew (PM) response, and the data were reassessed to determine the presence of QTL with pleiotropic effects. Phenotyping for leaf rust response was conducted in six environments at Baoding in Hebei province and Zhoukou in Henan province. The DH lines were genotyped using the wheat 55K SNP array and additional SSR markers. Eight QTLs for LR resistance were identified in the present study using the inclusive composite interval mapping (ICIM). The flanking sequences of all the SNP probes were subjected to BLAST against the Chinese Spring reference sequence to determine physical positions. Six LR resistance loci were contributed by Pingyuan 50, but QLr.hebau-2DS2 and QLr.hebau-3AL were from Mingxian 169. QLr.hebau-3DS, a new and stable QTL detected in all environments, explained 4.2–20.0% of the phenotypic variation. Another new QTL, QLr.hebau-3AL detected in four environments had a PVE (phenotypic variance explained) of 5.7–10.1%. QLr.hebau-1BL, QLr.hebau-2DS2, and QLr.hebau-7DL are new loci. Two QTLs showed potential pleiotropic effects on disease response. QTL on chromosome 7DL might be pleiotropic to LR and PM, and the QTL on 5DL might confer pleiotropic resistance to LR and YR. Kompetitive allele-specific PCR (KASP) markers for QLr.hebau-3AL and QLr.hebau-3DS were successfully developed and validated. The QTL identified with their flanking markers and in the present study could be useful for fine mapping, candidate gene mining, and marker-assisted selection (MAS).

The Kenyan wheat (Triticum aestivum L.) ‘Kenya Kongoni’ exhibits high levels of adult plant resistance (APR) to leaf rust (LR) and yellow rust (YR). We determined the genomic regions associated with LR and YR resistance in a population of 148 recombinant inbred lines generated from a cross between ‘Avocet-YrA’ and Kenya Kongoni. Field experiments to characterize APR to LR and YR were conducted in four and two Mexican or Uruguayan environments, respectively. A linkage map was constructed with 438 diversity arrays technology and 16 simple-sequence repeat markers by JoinMap 4.1 software. Genetic analyses showed that resistance to both rusts was determined by four to five APR genes, including Lr46/Yr29 and Sr2/Lr27/Yr30. Quantitative trait loci (QTL) analysis indicated that pleiotropic APR loci QYLr.cim-1BL corresponding to Lr46/Yr29 and QYLr.cim-7BL that is a putative novel QTL accounted for 5 to 57% and 12 to 35% of the phenotypic variation for resistance to LR and YR, respectively. These loci, in combination with another three LR QTL and two YR QTL, respectively, conferred high levels of resistance to both LR and YR in wheat under Mexican and Uruguayan environments. Among other detected QTL, QLr.cim-1DS, QLr.cim-2BL, and QYLr.icm-7BL may be new loci for APR to both rusts in common wheat.

Phenotyping of wheat stripe rust in genetic studies has traditionally relied on quantifying disease by means of the modified Cobb Scale. This approach requires scoring of disease severity and response type, either on flag leaves or on a whole plot basis. The use of spectral crop sensors in wheat phenotyping has raised the question of whether this objective methodology is suitable for detecting stripe rust resistance loci in genetic studies. An Avocet S X Francolin#1 recombinant inbred population developed at the International Maize and Wheat Improvement Center (Centro Internacional de Mejoramiento de Maiz y Trigo—acronym CIMMYT), and a Kariega X Avocet S doubled haploid population developed in South Africa, both with available genetic maps, were used in this study. Field trials for stripe rust evaluation of these populations were planted at Greytown, South Africa in 2013 and 2014. Severe and uniformly distributed infection of Puccinia striiformis race 6E22A+ occurred in both years. Populations and parents were phenotyped according to the modified Cobb Scale and with a handheld Trimble GreenSeeker® crop sensor (model HCS-100). The sensing device emits red and infrared light and measures the reflectance of each wavelength in terms of the normalized difference vegetation index (NDVI). In both years, NDVI detected four previously characterized quantitative trait loci (QTL) on chromosomes 1BL, 2BS, 3BS and 6AL in the Avocet S X Francolin#1 population. The same QTL were detected with visual estimates in 2013 but only 2BS was significant in 2014. In the Kariega X Avocet S population, all stripe rust traits, including NDVI, mapped to previously described QTL on chromosomes 2BS, 4AL and 7DS. Remote sensing of infection levels thus consistently detected the same QTL regions as described by using visual ratings in earlier studies, indicating that a crop sensor can be easily applied in genetic mapping of stripe rust resistance in wheat.

Powdery mildew resistance gene Pm55 was physically mapped to chromosome arm 5VS FL 0.60–0.80 of Dasypyrum villosum . Pm55 is present in T5VS·5AL and T5VS·5DL translocations, which should be valuable resources for wheat improvement. Powdery mildew caused by Blumeria graminis f. sp. tritici is a major wheat disease worldwide. Exploiting novel genes effective against powdery mildew from wild relatives of wheat is a promising strategy for controlling this disease. To identify novel resistance genes for powdery mildew from Dasypyrum villosum, a wild wheat relative, we evaluated a set of Chinese Spring-D. villosum disomic addition and whole-arm translocation lines for reactions to powdery mildew. Based on the evaluation data, we concluded that the D. villosum chromosome 5V controls post-seedling resistance to powdery mildew. Subsequently, three introgression lines were developed and confirmed by molecular and cytogenetic analysis following ionizing radiation of the pollen of a Chinese Spring-D. villosum 5V disomic addition line. A homozygous T5VS·5AL translocation line (NAU421) with good plant vigor and full fertility was further characterized using sequential genomic in situ hybridization, C-banding, and EST-STS marker analysis. A dominant gene permanently named Pm55 was located in chromosome bin 5VS 0.60–0.80 based on the responses to powdery mildew of all wheat-D. villosum 5V introgression lines evaluated at both seeding and adult stages. This study demonstrated that Pm55 conferred growth-stage and tissue-specific dependent resistance; therefore, it provides a novel resistance type for powdery mildew. The T5VS·5AL translocation line with additional softness loci Dina/Dinb of D. villosum provides a possibility of extending the range of grain textures to a super-soft category. Accordingly, this stock is a new source of resistance to powdery mildew and may be useful in both resistance mechanism studies and soft wheat improvement.

Chemotherapeutic agents (ChAs) are considered an integral part of current treatment protocols for the decontamination of titanium implants with peri-implantitis, based on their antimicrobial effect. Despite the proven antimicrobial effect of ChAs on titanium-bound biofilms, previous studies have elucidated an unexpected disassociation between bacterial reduction and biologically acceptable treatment outcomes. In this study, the authors hypothesize that ChAs residues alter titanium physicochemistry and thus compromise cellular response to decontaminated surfaces.Grit-blasted acid-etched titanium disks were contaminated with multispecies microcosm biofilms grown from in vivo peri-implant plaque samples. To simulate implant decontamination, the contaminated disks were burnished with 0.12% chlorhexidine, 20% citric acid, 24% EDTA/1.5% NaOCl, or sterile saline and assessed surface physicochemical properties. Sterile untreated surfaces were the controls. The biologic effects of decontamination were assessed via cell proliferation and differentiation assays.Bacterial counts after decontamination confirmed that the ChAs were antimicrobial. X-ray photoelectron spectroscopy invariably detected elemental contaminants associated with each ChA molecule or salt that significantly altered wettability compared with controls. Notably, all surfaces with ChA residues showed some cytotoxic effect compared with controls (P0.05). Increased cell counts were consistently found in the saline-treated group compared with chlorhexidine (P = 0.03). Interestingly, no association was found between antimicrobial effect and cell counts (P0.05).ChA-specific residues left on the titanium surfaces altered titanium physical properties and adversely affected the osteoblastic response irrespective of their observed antimicrobial effect. Chlorhexidine may compromise the biocompatibility of titanium surfaces, and its use is not recommended to detoxify implants. Sterile saline, citric acid, and NaOCl-EDTA may be proposed for use in the treatment of peri-implantitis. Contrary to previous studies that recommended the selection of ChAs for the decontamination of titanium implants according to their antimicrobial effects, the present study demonstrated that the restoration of the biocompatibility of contaminated titanium surfaces is also contingent on the preservation of titanium material properties.

A complete set wheat-R. ciliaris disomic addition lines (DALs) were characterized and the homoeologous groups and genome affinities of R. ciliaris chromosomes were determined. Wild relatives are rich gene resources for cultivated wheat. The development of alien addition chromosome lines not only greatly broadens the genetic diversity, but also provides genetic stocks for comparative genomics studies. Roegneria ciliaris (genome ScScYcYc), a tetraploid wild relative of wheat, is tolerant or resistant to many abiotic and biotic stresses. To develop a complete set of wheat-R. ciliaris disomic addition lines (DALs), we undertook a euplasmic backcrossing program to overcome allocytoplasmic effects and preferential chromosome transmission. To improve the efficiency of identifying chromosomes from Sc and Yc, we established techniques including sequential genomic in situ hybridization/fluorescence in situ hybridization (FISH) and molecular marker analysis. Fourteen DALs of wheat, each containing one pair of R. ciliaris chromosomes pairs, were characterized by FISH using four repetitive sequences [pTa794, pTa71, RcAfa and (GAA)10] as probes. One hundred and sixty-two R. ciliaris-specific markers were developed. FISH and marker analysis enabled us to assign the homoeologous groups and genome affinities of R. ciliaris chromosomes. FHB resistance evaluation in successive five growth seasons showed that the amphiploid, DA2Yc, DA5Yc and DA6Sc had improved FHB resistance, indicating their potential value in wheat improvement. The 14 DALs are likely new gene resources and will be phenotyped for more agronomic performances traits.

To develop a method for quantifying leakage in composite resin restorations after curing, using non-destructive X-ray micro-computed tomography (micro-CT) and image segmentation.Class-I cavity preparations were made in 20 human third molars, which were divided into 2 groups. Group I was restored with Z100 and Group II with Filtek LS. Micro-CT scans were taken for both groups before and after they were submerged in silver nitrate solution (AgNO3 50%) to reveal any interfacial gap and leakage at the tooth restoration interface. Image segmentation was carried out by first performing image correlation to align the before- and after-treatment images and then by image subtraction to isolate the silver nitrate penetrant for precise volume calculation. Two-tailed Student's t-test was used to analyze the results, with the level of significance set at p0.05.All samples from Group I showed silver nitrate penetration with a mean volume of 1.3 ± 0.7mm(3). In Group II, only 2 out of the 10 restorations displayed infiltration along the interface, giving a mean volume of 0.3 ± 0.3mm(3). The difference between the two groups was statistically significant (p0.05). The infiltration showed non-uniform patterns within the interface.We have developed a method to quantify the volume of leakage using non-destructive micro-CT, silver nitrate infiltration and image segmentation. Our results confirmed that substantial leakage could occur in composite restorations that have imperfections in the adhesive layer or interfacial debonding through polymerization shrinkage. For the restorative systems investigated in this study, this occurred mostly at the interface between the adhesive system and the tooth structure.

Stripe rust, caused by Puccinia striiformis f. sp. tritici W., is a devastating disease of wheat worldwide. A new stripe rust resistance gene with moderate seedling and adult plant resistance was mapped using an F5 recombinant inbred line (RIL) population developed from the cross of the resistant parent ‘Almop’ with the susceptible parent ‘Avocet’. The parents and RILs were phenotyped for seedling stripe rust response variation in a greenhouse and in field trials at Toluca, Mexico for 2 years. Almop showed moderate levels of resistance at both seedling and adult plant stages compared with the highly susceptible response of Avocet. The distribution of homozygous resistant, homozygous susceptible, and segregating RILs conformed to segregation at a single locus. Seedlings and adult plant responses were correlated, indicating that the same gene conferred resistance at both stages. A bulk segregant analysis approach with widely distributed simple sequence repeat (SSR) markers mapped the resistance gene to the distal region of the long arm of chromosome 4A. The SSR marker wmc776 cosegregated with this gene, whereas markers wmc219 and wmc313 were tightly linked and both located at 0.6 centimorgans. The resistance locus was designated Yr60.

Leaf rust (LR), caused by Puccinia triticina , is one of the most widespread diseases of common wheat ( Triticum aestivum L.) worldwide. The LR resistance gene LrBi16 has been mapped on chromosome arm 7BL in Chinese wheat cultivar Bimai 16 and was closely linked to SSR loci Xcfa2257 and Xgwm344 with genetic distances of 2.8 cM and 2.9 cM, respectively. In the present study, a total of 304 AFLP primer pairs were used to screen Bimai 16 and Thatcher and resistant and susceptible DNA bulks. The polymorphic AFLP marker P-ATT/M-CGC 173 bp was used to genotype F 2 and F 3 populations to identify markers more closely linked to LrBi16 . Marker P-ATT/M-CGC 173 bp was tightly linked to LrBi16 with a genetic distance of 0.5 cM. As LrBi16 was mapped near the Lr14a locus, 809 F 2 plants from the Bimai 16/RL6013 ( Lr14a ) cross were inoculated with the Pt pathotype FHNQ to test the allelism of Lr14a and LrBi16 . All of the F 2 plants were resistant to FHNQ (IT between; and 2), suggesting that Lr14a and LrBi16 are allelic.