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Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), poses a severe threat to wheat yield and quality worldwide. Rapid identification and the accurate transference of effective resistance genes are important to the development of resistant cultivars and the sustainable control of this disease. In the present study, the wheat line AL11 exhibited high levels of resistance to powdery mildew at both the seedling and adult plant stages. Genetic analysis of the AL11 × Shixin 733 mapping population revealed that its resistance was controlled by a single dominant gene, tentatively designated PmAL11. Using bulked segregant RNA-Seq (BSR-Seq) and molecular marker analysis, PmAL11 was mapped to the Pm5 locus on chromosome 7B where it co-segregated with the functional marker Pm5e-KASP. Sequence alignment analysis revealed that the Pm5e-homologous sequence in AL11 was identical to the reported recessive gene Pm5e in wheat landrace Fuzhuang 30. It appears that PmAL11 was most probably Pm5e but it was mediated by a dominant inheritance pattern, so it should provide a valuable resistance resource for both genetic study and wheat breeding. To efficiently use and trace PmAL11 in breeding, a new kompetitive allele-specific PCR (KASP) marker AL11-K2488 that co-segregated with this gene was developed and was confirmed to be applicable in the different wheat backgrounds, thus promoting its use in the marker-assisted selection (MAS) of PmAL11.

Rye (Secale cereale L., RR) contains valuable genes for wheat improvement. However, most of the rye resistance genes have not been successfully used in wheat cultivars. Identification of new rye resistance genes and transfer these genes to wheat by developing small fragment translocation lines will make these genes more usable for wheat breeding. In this study, a broad-spectrum powdery mildew resistance gene PmW6RS was located on rye chromosome arm 6RS using a new set of wheat-rye disomic and telosomic addition lines. To further study and use PmW6RS, 164 wheat-rye 6RS translocation lines were developed by 60Coγ ray irradiation. Seedling and adult stage powdery mildew resistance analysis showed that 106 of the translocation lines were resistant. Physical map of 6RS was constructed using the 6RS translocation and deletion lines, and PmW6RS was located in the 6RS-0.58-0.66-bin, flanked by markers X6RS-3 and X6RS-10 corresponding to the physical interval of 50.23-68.61 Mb, and was 18.38 Mb in Weining rye genome. A total of 23 resistance-related genes were annotated. Nine markers co-segregate with the 6RS-0.58-0.66-bin, which can be used to rapidly trace the 6RS fragment carrying PmW6RS. Small fragment translocation lines with powdery mildew resistance were backcrossed with wheat major cultivars and 39 homozygous 6RS small fragment translocation lines were obtained. In conclusion, this study not only provide novel gene source and germplasms for wheat resistance breeding, but also laid solid foundation for further cloning of PmW6RS.

IntroductionIdentification of stable major quantitative trait loci (QTLs) for yield-related traits is important for yield potential improvement in wheat breeding.MethodsIn the present study, we genotyped a recombinant inbred line (RIL) population using the Wheat 660K SNP array and constructed a high-density genetic map. The genetic map showed high collinearity with the wheat genome assembly. Fourteen yield-related traits were evaluated in six environments for QTL analysis.Results and DiscussionA total of 12 environmentally stable QTLs were identified in at least three environments, explaining up to 34.7% of the phenotypic variation. Of these, QTkw-1B.2 for thousand kernel weight (TKW), QPh-2D.1 (QSl-2D.2/QScn-2D.1) for plant height (PH), spike length (SL) and spikelet compactness (SCN), QPh-4B.1 for PH, and QTss-7A.3 for total spikelet number per spike (TSS) were detected in at least five environments. A set of Kompetitive Allele Specific PCR (KASP) markers were converted based on the above QTLs and used to genotype a diversity panel comprising of 190 wheat accessions across four growing seasons. QPh-2D.1 (QSl-2D.2/QScn-2D.1), QPh-4B.1 and QTss-7A.3 were successfully validated. Compared with previous studies, QTkw-1B.2 and QPh-4B.1 should be novel QTLs. These results provided a solid foundation for further positional cloning and marker-assisted selection of the targeted QTLs in wheat breeding programs.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a devastating disease that seriously threatens the wheat yield and quality. To control this disease, host resistance is the preferred measure. However, wheat breeding is a complex process with elusive exchange and recombination of the traits from their parents. Increased resistance often leads to a decline in other key traits, such as yield and quality. Developing breakthrough germplasms with harmonious powdery mildew resistance and other key breeding traits is attractive in wheat breeding. In this study, we developed an ideal wheat breeding line AL46 that pyramided its hexaploid triticale parent-derived desirable yield traits and its wheat parent-derived powdery mildew resistance gene Pm2. Sequential genomic in situ hybridization (GISH), multicolor GISH (mc-GISH), multicolor fluorescence in situ hybridization (mc-FISH), and molecular marker analyses revealed that AL46 was a wheat-rye T1RS·1BL translocation line. Genetic analysis combined with function marker detection and sequence alignment were used to confirm that AL46 carried the Pm2 gene. Then, we evaluated the powdery mildew resistance and comprehensive traits of AL46, and just as we designed, AL46 showed harmonious powdery mildew resistance and some key breeding traits. This study not only developed an ideal wheat germplasm resource, but also provided a successful example for pyramiding breeding, which could be a promising direction for wheat improvement in the future.

Powdery mildew and leaf rust, caused by Blumeria graminis f. sp. tritici and Puccinia triticina, respectively, are widespread diseases of wheat worldwide. The use of resistant cultivars is considered the most economical, environment-friendly, and effective method to control these diseases. In the present study, a collection of 2,978 wheat accessions consisting of 1,394 advanced breeding lines, 1,078 Chinese cultivars, 291 introduced cultivars, 132 lines containing alien chromosomes, and 83 landraces was tested for reactions to powdery mildew and leaf rust. The results indicated that 659 wheat accessions (22.1%) were highly resistant to a widely prevalent B. graminis f. sp. tritici isolate, E09, at the seedling stage, and 390 were consistently resistant to the mixture of B. graminis f. sp. tritici isolates at the adult plant stage. Meanwhile, 63 accessions (2.1%) were highly resistant to leaf rust at the adult plant stage, of which 54 were resistant to a predominant and highly virulent P. triticina race, THTT, at the seedling stage. Notably, 17 accessions were resistant to both powdery mildew and leaf rust. To detect known genes for resistance to powdery mildew and leaf rust, these accessions were tested with gene-specific or tightly linked markers for seven powdery mildew genes (Pm genes; Pm2, Pm4, Pm5, Pm6, Pm8, Pm21, and Pm24) and 10 Lr genes (Lr1, Lr9, Lr10, Lr19, Lr20, Lr24, Lr26, Lr34, Lr37, and Lr46). Of the 659 powdery mildew-resistant accessions, 328 might carry single Pm genes and 191 carry combined Pm genes. Pm2 was detected at the highest frequency of 59.6%, followed by Pm8, Pm6, Pm21, Pm4, and Pm5, whereas Pm24 was not detected. In addition, 139 accessions might contain unknown Pm genes different from those tested in this study. In the 63 accessions resistant to leaf rust, four leaf rust genes (Lr genes; Lr1, Lr10, Lr26, and Lr34) were detected in 41 accessions singly or in combination, whereas six genes (Lr9, Lr19, Lr20, Lr24, Lr37, and Lr46) were not detected. Twenty-two accessions might contain unknown Lr genes different from those tested in this study. This study not only provided important information for rationally distributing resistance genes in wheat breeding programs, but also identified resistant germplasm that might have novel genes to enrich the diversity of resistance sources.

Background Plant height (PH), spike length (SL) and spike compactness (SCN) are important agronomic traits in wheat due to their strong correlations with lodging and yield. Thus, dissection of their genetic basis is essential for the improvement of plant architecture and yield potential in wheat breeding. The objective of this study was to map quantitative trait loci (QTL) for PH, SL and SCN in a recombinant inbred line (RIL) population derived from the cross ‘PuBing3228 × Gao8901’ (PG-RIL) and to evaluate the potential values of these QTL to improve yield. Results In the current study, Five, six and ten stable QTL for PH, SL, and SCN, respectively, were identified in at least two individual environments. Five major QTL QPh.cas-5A.3, QPh.cas-6A, QSl.cas-6B.2, QScn.cas-2B.2 and QScn.cas-6B explained 5.58–25.68% of the phenotypic variation. Notably, two, three and three novel stable QTL for PH, SL and SCN were identified in this study, which could provide further insights into the genetic factors that shape PH and spike morphology in wheat. Conditional QTL analysis revealed that QTL for SCN were mainly affected by SL. Moreover, a Kompetitive Allele Specific PCR (KASP) marker tightly linked to stable major QTL QPh.cas-5A.3 was developed and verified using the PG-RIL population and a natural population. Conclusions Twenty-one stable QTL related to PH, SL, and SCN were identified. These stable QTL and the user-friendly marker KASP8750 will facilitate future studies involving positional cloning and marker-assisted selection in breeding.

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a destructive disease of wheat throughout the world. Host resistance is considered the most sustainable way to control this disease. Powdery mildew resistance gene Pm2b was mapped to the same genetic interval with Pm2a and PmCH1357 cloned previously, but showed different resistance spectra from them, indicating that they might be caused by different resistance genes or alleles. In this study, Pm2b was delimited to a 1.64 Mb physical interval using a large segregating population containing 4,354 F2:3 families of resistant parent KM2939 and susceptible cultivar Shimai 15. In this interval, TraesCS5D03G0111700 encoding the coiled-coil nucleotide-binding site leucine-rich repeat protein (CC-NBS-LRR) was determined as the candidate gene of Pm2b. Silencing by barley stripe mosaic virus-induced gene silencing (BSMV-VIGS) technology and two independent mutants analysis in KM2939 confirmed the candidate gene TraesCS5D03G0111700 was Pm2b. The sequence of Pm2b was consistent with Pm2a/PmCH1357. Subcellular localization showed Pm2b was located on the cell nucleus and plasma membrane. Pm2b had the highest expression level in leaves and was rapidly up-regulated after inoculating with Bgt isolate E09. The yeast two-hybrid (Y2H) and luciferase complementation imaging assays (LCI) showed that PM2b could self-associate through the NB domain. Notably, we identified PM2b interacting with the transcription factor TaWRKY76-D, which depended on the NB domain of PM2b and WRKY domain of TaWRKY76-D. TaWRKY76-D negatively regulated the resistance to powdery mildew in wheat. The specific KASP marker K529 could take the advantage of high-throughput and high-efficiency for detecting Pm2b and be useful in molecular marker assisted-selection breeding. In conclusion, cloning and disease resistance mechanism analysis of Pm2b provided an example to emphasize a need of the molecular isolation of resistance genes, which has implications in marker assisted wheat breeding.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive fungal disease of wheat throughout the world. Utilization of effective powdery mildew resistance genes and cultivars is considered as the most economic, efficient, and environmental-friendly method to control this disease. Synthetic hexaploid wheat (SHW), which was developed through hybridization of diploid Aegilops and tetraploid wheat, is a valuable genetic resource for resistance to powdery mildew. SHW line YAV249 showed high levels of resistance to powdery mildew at both the seedling and adult stages. Genetic analysis indicated that the resistance was controlled by a single dominant gene, temporarily designated PmYAV. Bulked segregant analysis with wheat 660K single nucleotide polymorphism (SNP) array scanning and marker analysis showed that PmYAV was located on chromosome 2AL and flanked by markers Xgdm93 and Xwgrc763, respectively, with genetic distances of 0.8 cM and 1.2 cM corresponding to a physic interval of 1.89 Mb on the Chinese Spring reference genome sequence v1.0. Sequence alignment analysis demonstrated that the sequence of PmYAV was consistent with that of Pm4a but generated an extra splicing event. When inoculated with different Bgt isolates, PmYAV showed a significantly different spectrum from Pm4a, hence it might be a new resistant resource for improvement of powdery mildew resistance. The flanked markers GDM93 and WGRC763, and the co-segregated markers BCD1231 and JS717/JS718 were confirmed to be easily performed in marker-assisted selection (MAS) of PmYAV. Using MAS strategy, PmYAV was transferred into the commercial cultivar Kenong 199 (KN199) and a wheat line YK13 was derived at generation BC3F3 from the population of YAV249/4*KN199 due to its excellent agronomic traits and resistance to powdery mildew. In conclusion, an alternative splicing variant of Pm4 was identified in this study, which informed the regulation of Pm4 gene function.

Developing effective and durable host plant resistance is crucial for controlling powdery mildew, a devastating disease caused by Blumeria graminis f. sp. tritici (Bgt). In the present study, we dissected the genetic basis of the adult plant resistance to powdery mildew using a recombinant inbred line (RIL) composed of 176 F9 RILs population derived from a cross between PuBing 3228 (P3228) and susceptible cultivar Gao 8901. P3228 exhibits stable adult-plant resistance to powdery mildew in the field over consecutive years. We identified two QTLs on chromosomes 7DS (QPm.cas-7D) and 1AL (QPm.cas-1A) contributed by P3228, and one QTL on 3DS (QPm.cas-3D) contributed by Gao 8901, which could explain 65.44%, 3.45%, and 2.18% of the phenotypic variances, respectively. By analyzing the annotated genes in the 1.168 Mb physical interval of the major QTL QPm.cas-7D, we locked a previously cloned adult-plant resistance gene Pm38 that was most probably the candidate gene of QPm.cas-7D. Sequence alignment analysis revealed that the candidate gene of QPm.cas-7D in P3228 was identical to the reported Pm38 sequence. Two haplotypes QPm-7D-R and QPm-7D-S were identified in the whole Pm38 genomic regions between P3228 and Gao 8901. To apply QPm.cas-7D in wheat breeding, we developed a kompetitive allele-specific PCR (KASP) marker Kasp5249 that is closely linked with these haplotypes. It is worth mentioning that the QPm-7D-R haplotype significantly decreased TKW and underwent negative selection for higher yields in China wheat breeding. In this study, we identified a major QTL QPm.cas-7D and revealed the relationship between its resistance and yield, which could be beneficial for further applications in wheat disease resistance and high-yield breeding.

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Wheat-rye T1RS·1BL translocations have been widely used worldwide in wheat production for multiple disease resistance and superior yield traits. However, many T1RS·1BL translocations have successively lost their resistance to pathogens due to the coevolution of pathogen virulence with host resistance. Because of the extensive variation in rye (Secale cereale L.) as a naturally cross-pollinating relative of wheat, it still has promise to widen the variation of 1RS and to fully realize its application value in wheat improvement. In the present study, the wheat-rye breeding line R2207 was characterized by comprehensive analyses using genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization with multiple probes, multicolor GISH, and molecular marker analysis, and then was proven to be a cytogenetically stable wheat-rye T1RS·1BL translocation line. Based on the disease responses to different isolates of powdery mildew and genetic analysis, R2207 appears to possess a novel variation for resistance, which was confirmed to be located on the rye chromosome arm 1RS. Line R2207 also exhibited high levels of resistance to stripe rust at both seedling and adult stages, as well as enhanced agronomic performance, so it has been transferred into a large number of commercial cultivars using an efficient 1RS-specific kompetitive allele specific PCR marker for marker-assisted selection.

Rye (Secale cereale L.), a naturally cross-pollinating relative of wheat, is a tertiary gene donor and of substantial value in wheat improvement. Wheat powdery mildew is caused by Blumeria graminis f. sp. tritici (Bgt), which seriously affects yield and quality worldwide. Identifying and transferring new, effective resistance genes against powdery mildew from rye is important for wheat breeding. The current study developed a wheat-rye line YT2 resistant to powdery mildew by crossing, backcrossing, and self-pollination for multiple generations between octoploid triticale 09R2-100 and common wheat cultivar Shixin 616. YT2 was confirmed to be a 6R disomic addition and T1RS⋅1BL translocation line by genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization (mc-FISH), multicolor-GISH (mc-GISH), and molecular marker analyses. Disease responses to different Bgt isolates and genetic analysis showed that the powdery mildew resistance gene of YT2 was derived from the rye chromosome 6R of 09R2-100, which differed from the previously reported Pm genes from rye including Pm20 on 6RL. Resistance phenotype of different translocation lines and deletion lines derived from YT2 combined with newly developed 6RL-specific markers analysis suggested that the powdery mildew resistance gene of YT2 was localized to the region in chromosome 6RL: 890.09–967.51 Mb and flanked by markers XM189 and X4M19, corresponding to the reference genome of Weining rye. Therefore, YT2 could be used as a promising bridging parent for wheat disease resistance improvement.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a devastating disease of wheat that seriously affects yield and quality worldwide. Because of the extensive growth of wheat cultivars with homogeneous genetic background, exploring novel resistant resources from wheat relatives has become important for increasing the genetic diversity of wheat. Rye (Secale cereale) is a wheat relative possessing abundant resistance genes because of its high variation. Wheat line AL69, resistant to powdery mildew, was developed by crossing, backcrossing, and self-pollination for multiple generations between hexaploid triticale Zhongsi 237 and common wheat cultivar Zimai 17. Through genomic in situ hybridization (GISH) and multicolor fluorescence in situ hybridization (FISH), nondenaturing FISH, multicolor GISH, and selection with specific molecular markers, AL69 was determined to be a wheat-rye 2R (2D) disomic substitution line. Testing with different B. graminis f. sp. tritici isolates and genetic analysis showed that the all-stage resistance (also called seedling resistance) of AL69 was conferred by the cataloged powdery mildew resistance gene Pm4b derived from Zimai 17, and its adult-plant resistance was derived from the alien chromosome 2R of Zhongsi 237, which was found to be different from the previously reported rye-derived Pm genes, including Pm7 on 2RL. In addition, AL69 showed improved spike number per plant, spike length, fertile spikelet number per spike, kernel number per spike, and grain yield per plant compared with its wheat parent Zimai 17. An elite line S251 combining powdery mildew resistance with excellent agronomic performance was selected from the progenies of AL69 and wheat cultivar Jimai 22. Therefore, AL69 has two types of resistance genes to powdery mildew and improved agronomic traits through pyramiding and thus can be used as a promising genetic stock for wheat breeding.

Summary Kernel size is an important trait determining cereal yields. In this study, we cloned and characterized TaDA1, a conserved negative regulator of kernel size in wheat (Triticum aestivum). The overexpression of TaDA1 decreased the size and weight of wheat kernels, while its down‐regulation using RNA interference (RNAi) had the opposite effect. Three TaDA1‐A haplotypes were identified in Chinese wheat core collections, and a haplotype association analysis showed that TaDA1‐A‐HapI was significantly correlated with the production of larger kernels and higher kernel weights in modern Chinese cultivars. The haplotype effect resulted from a difference in TaDA1‐A expression levels between genotypes, with TaDA1‐A‐HapI resulting in lower TaDA1‐A expression levels. This favourable haplotype was found having been positively selected during wheat breeding over the last century. Furthermore, we demonstrated that TaDA1‐A physically interacts with TaGW2‐B. The additive effects of TaDA1‐A and TaGW2‐B on kernel weight were confirmed not only by the phenotypic enhancement arising from the simultaneous down‐regulation of TaDA1 and TaGW2 expression, but also by the combinational haplotype effects estimated from multi‐environment field data from 348 wheat cultivars. A comparative proteome analysis of developing transgenic and wild‐type grains indicated that TaDA1 and TaGW2 are involved in partially overlapping but relatively independent protein regulatory networks. Thus, we have identified an important gene controlling kernel size in wheat and determined its interaction with other genes regulating kernel weight, which could have beneficial applications in wheat breeding.

Additional file 2: Supplementary file 2.

Chinese wheat landrace Dahongtou was resistant to 35 of 38 tested Chinese isolates of Blumeria graminis f. sp. tritici at the seedling stage. Genetic analysis of the F2 populations and their derived F2:3 families of crosses of Dahongtou with the susceptible varieties Mingxian 169 and Huixianhong indicated that the resistance of Dahongtou to B. graminis f. sp. tritici isolate E09 was conferred by a single recessive gene, tentatively designated as pmDHT. The gene was mapped to chromosome arm 7BL and flanked by markers Xwmc526/XBE443877 and Xgwm611/Xwmc511 at genetic distances of 0.8 and 0.3 cM, respectively. The chromosomal position of pmDHT was similar to the multi-allelic Pm5 locus on 7BL. Allelism tests with crosses of Dahongtou with Fuzhuang 30 (Pm5e) and Xiaobaidong (mlxbd) indicated that pmDHT was allelic to both Pm5e and mlxbd. However, pmDHT showed a different pattern of resistance to the 38 B. graminis f. sp. tritici isolates compared with wheat lines with Pm5a, Pm5b, Pm5e, mlxbd, and PmHYM and also differed from PmSGA. Thus, pmDHT was identified most likely as a new allele or at least a closely linked gene of the Pm5 locus. This gene can be transferred into susceptible wheat cultivars/lines and pyramided with other resistance genes through marker-assisted selection to improve powdery mildew resistance.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat worldwide. The disease is most preferably managed by developing and growing cultivars with high-level, durable resistance. To achieve high-level and long-lasting resistance, we developed a wheat line, RIL-Yr64/Yr15, by pyramiding Yr64 and Yr15, both on the chromosome 1BS and providing high resistance to all tested Pst races. To validate RIL-Yr64/Yr15 possessing both genes, we crossed it to Avocet S (AvS). The F4 RILs from this cross were phenotyped with Pst races under controlled greenhouse conditions and also under natural Pst infection in the field. The population was genotyped with SSR markers previously reported to be linked to the resistance gene loci and with additional SSR and SNP-KASP markers along chromosome 1B. Both phenotype and genotype data confirmed the copresence of Yr64 and Yr15 in RIL-Yr64/Yr15, and the high-resolution linkage map dissected the chromosomal regions and traced their origins. New lines possessing these genes were selected from the F5 population of cross AvS × RIL-Yr64/Yr15 by marker-assisted selection. These lines with the two highly effective genes should be more useful than individual gene lines for developing high-level, durable resistant wheat cultivars.

BackgroundKernel weight and morphology are important traits affecting cereal yields and quality. Dissecting the genetic basis of thousand kernel weight (TKW) and its related traits is an effective method to improve wheat yield.ResultsIn this study, we performed quantitative trait loci (QTL) analysis using recombinant inbred lines derived from the cross ‘PuBing3228 × Gao8901’ (PG-RIL) to dissect the genetic basis of kernel traits. A total of 17 stable QTLs related to kernel traits were identified, notably, two stable QTLsQTkw.cas-1A.2andQTkw.cas-4Aexplained the largest portion of the phenotypic variance for TKW and kernel length (KL), and the other two stable QTLsQTkw.cas-6A.1andQTkw.cas-7D.2contributed more effects on kernel width (KW). Conditional QTL analysis revealed that the stable QTLs for TKW were mainly affected by KW. The QTLsQTkw.cas-7D.2andQKw.cas-7D.1associated with TKW and KW were delimited to the physical interval of approximately 3.82 Mb harboring 47 candidate genes. Among them, the candidate geneTaFT-D1had a 1 bp insertions/deletion (InDel) within the third exon, which might be the reason for diversity in TKW and KW between the two parents. A Kompetitive Allele-Specific PCR (KASP) marker ofTaFT-D1allele was developed and verified by PG-RIL and a natural population consisted of 141 cultivar/lines. It was found that the favorableTaFT-D1 (G)-allelehas been positively selected during Chinese wheat breeding. Thus, these results can be used for further positional cloning and marker-assisted selection in wheat breeding programs.ConclusionsSeventeen stable QTLs related to kernel traits were identified. The stable QTLs for thousand kernel weight were mainly affected by kernel width.TaFT-D1could be the candidate gene for QTLsQTkw.cas-7D.2andQKw.cas-7D.1.

Additional file 2: Table S2. Detection of 1RS translocations in 161 wheat cultivars/ lines.

Additional file 1: Table S1. Primer sequences of the 404 molecular markers specific for 14 rye chromosome arms.

A wheat-rye 4R chromosome disomic addition line with resistances to powdery mildew, stripe rust, sharp eyespot and high kernel number per spike was developed and characterized by molecular cytogenetic method as novel resistant germplasm. Rye (Secale cereale L.), a close relative of common wheat, is an important and valuable gene donor with multiple disease resistance for wheat improvement. However, resistance genes derived from rye have successively lost resistance to pathogens due to the coevolution of pathogen virulence and host resistance. Development and identification of new effective resistance gene sources from rye therefore are of special importance and urgency. In the present study, a wheat-rye line WR35 was produced through distant hybridization, embryo rescue culture, chromosome doubling and backcrossing. WR35 was then proven to be a new wheat-rye 4R disomic addition line using sequential GISH (genomic in situ hybridization), mc-FISH (multicolor fluorescence in situ hybridization) and ND-FISH (non-denaturing FISH) with multiple probes, mc-GISH (multicolor GISH), rye chromosome arm-specific marker analysis and SLAF-seq (specific-locus amplified fragment sequencing) analysis. At the adult stage, WR35 exhibited high levels of resistance to the powdery mildew (Blumeria graminis f. sp. tritici, Bgt) and stripe rust (Puccinia striiformis f. sp. tritici, Pst) pathogens prevalent in China, and a highly virulent isolate of Rhizoctonia cerealis, the cause of wheat sharp eyespot. At the seedling stage, it was highly resistant to 22 of 23 Bgt isolates and four Pst races. Based on its disease responses to different pathogen isolates, WR35 may possess resistance gene(s) for powdery mildew, stripe rust and sharp eyespot, which differed from the known resistance genes from rye. In addition, WR35 was cytologically stable and produced high kernel number per spike. Therefore, WR35 with multi-disease resistances and desirable agronomic traits should serve as a promising bridging parent for wheat chromosome engineering breeding.

Rye (Secale cereale L.) is an important gene donor for wheat improvement because of its many valuable traits, especially disease resistance. Development of novel wheat-rye translocations with disease resistance can contribute to transferring resistance into common wheat. In a previous study, a wheat-rye T4BL·4RL and T7AS·4RS translocation line (WR41-1) was developed by distant hybridization, and it was speculated that its resistance to powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), was derived from rye based on pedigree analysis. To make accurate use of chromosome 4R in wheat improvement, a set of new 4R translocations involving different arm translocations (e.g., 4RS monosomic, 4RL monosomic, 4RL disomic, 4RS monosomic plus 4RL monosomic, 4RS monosomic plus 4RL disomic, and 4RS disomic plus 4RL disomic translocations) was developed from crosses with common wheat. Those translocations were characterized by genomic in situ hybridization and expressed sequence tag simple sequence repeat marker analysis. To confirm the source of powdery mildew resistance, the translocation plants were tested against Bgt isolate E09. The results indicated that all translocations with 4RL were resistant at all tested growth stages, whereas those with only 4RS translocation or no alien translocation were susceptible. This further indicated that the powdery mildew resistance of WR41-1 was derived from the alien chromosome arm 4RL. To effectively use 4RL resistance in wheat improvement, two competitive allele-specific PCR markers specific for chromosome arm 4RL were developed to detect the alien chromosome in the wheat genome. These new translocation lines with diagnostic markers can efficiently serve as important bridges for wheat improvement.

Chinese wheat landrace Dahongtou was resistant to 35 of 38 tested Chinese isolates of

Wheat (Triticum aestivum) genotypes with rye (Secale cereale) 1RS chromosomal translocations are widely used in wheat breeding programs because 1RS carries genes for resistance to several diseases. However, some of the pathogens have evolved into new races that overcome the resistance due to extensive use of cultivars with the resistance genes from rye. Therefore, identification and deployment of new resistance sources with desirable agronomic characteristics are important and urgent. We have used winter rye cultivar German White as a source of genes for desirable traits in wheat improvement. A new genotype named WR04-32 was produced through hybridization and chromosome manipulation between common winter wheat cultivar Xiaoyan 6 and German White. This genotype was highly resistant to a wide spectrum of the wheat stripe rust (Puccinia striiformis f. sp. tritici) and powdery mildew (Blumeria graminis f. sp. tritici) pathotypes prevalent in China. The polymerase chain reaction (PCR) result using EST-STS (expressed sequence tag-site tagged sequence) marker STSWE126 specific to 1RS confirmed 1RS in WR04-32, and it was further proved to be a wheat-rye T2BL·1RS translocation line using sequential genomic in situ hybridization (GISH) and multicolor fluorescence in situ hybridization (FISH) with probes pAs1 and pSc119.2 (or pHvG38). In addition to its resistance to stripe rust and powdery mildew, WR04-32 was genetically stable and had desirable agronomic traits, making it a desirable germplasm for wheat breeding.

Exploitation of host resistance is important for controlling powdery mildew of wheat (Triticum aestivum L.). In this study, a wheat-Thinopyrum intermedium introgression line, designated WE99, conferred seedling resistance to 47 of 49 Blumeria graminis f. sp. tritici isolates. Genetic analysis demonstrated that the resistance segregation deviated significantly from a single gene Mendelian ratio. However, marker analysis indicated that only a single recessive resistance gene, temporarily designated PmWE99, conferred powdery mildew resistance (Pm). PmWE99 was mapped to chromosome arm 2BS and linked to the three simple-sequence repeat markers Gwm148, Gwm271, and Barc55. Using race spectrum analysis, PmWE99 was shown to be significantly different from the documented genes Pm42 and MlIW170 located on chromosome arm 2BS and, thus, appeared to be a new Pm gene. Examination of the genotype frequencies in the F2:3 families showed that a genetic variation in the PmWE99 interval that favored the transmission of the WE99 allele could be the cause of the deviated segregation. Further investigation revealed that the abnormal segregation only occurred at the PmWE99 interval and was not common at other loci in this population. Identification of PmWE99 will increase the diversity of the Pm genes for wheat improvement.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a serious disease of wheat (Triticum aestivum L.) throughout the world. Host resistance is the most effective and preferred means for managing this disease. Line 10V-2, a wheat breeding line with superior agronomic performance, shows broad-spectrum seedling resistance to powdery mildew. Genetic analysis demonstrated that its resistance was controlled by a single dominant gene, tentatively designated Pm10V-2. This gene was localized near the documented Pm2 locus on chromosome 5DS using the simple sequence repeat (SSR) marker Cfd81. To saturate the marker map of Pm10V-2, more markers were developed using bulked segregant RNA-Seq. Two single-nucleotide polymorphism (SNP) markers (Swgi047 and Swgi064), three expressed sequence tag markers (Swgi007, Swgi029, and Swgi038), and one SSR marker (Swgi066) were polymorphic between the resistant and susceptible bulks and showed tightly linked to the Pm10V-2 gene. Pm10V-2 was flanked by the new developed markers Swgi064 and Swgi066 at genetic distances of 0.4 and 1.2 centimorgans (cM), respectively, and cosegregated with Swgi007 and Swgi038. The homologous sequence of Pm2a was cloned from 10V-2 based on a recent study. Although the sequence cloned from 10V-2 was completely identical to that of the reported Pm2a-related gene, they did not cosegregate but were separated at a genetic distance of 1.6 cM, indicating that Pm10V-2 was different from the reported of Pm2a-related gene. When inoculated with multiple B. graminis f. sp. tritici isolates, Pm10V-2 had a significantly different resistance spectrum from Pm2a and other powdery mildew (Pm) resistance genes at or near the Pm2 locus. Therefore, Pm10V-2 may be a new Pm2 allele or Pm2-linked gene. To use Pm10V-2 in marker-assisted selection (MAS) breeding, seven markers applicable for MAS were confirmed, including three newly developed markers (Swgi029, Swgi038, and Swgi064) in the present work. Using these markers, a great number of resistant lines with desirable agronomic performance were selected from crosses involving 10V-2, including the breeding line KM5016, which has been entered in the Regional trials in Hebei Province, China.

Wheat spike development is a coordinated process of cell proliferation and differentiation with distinctive phases and architecture changes. However, the dynamic alteration of gene expression in this process remains enigmatic. Here, we characterized and dissected bread wheat spike into six developmental stages, and used genome-wide gene expression profiling, to investigate the underlying regulatory mechanisms. High gene expression correlations between any two given stages indicated that wheat early spike development is controlled by a small subset of genes. Throughout, auxin signaling increased, while cytokinin signaling decreased. Besides, many genes associated with stress responses highly expressed during the double ridge stage. Among the differentially expressed genes (DEGs), were identified 375 transcription factor (TF) genes, of which some homologs in rice or Arabidopsis are proposed to function in meristem maintenance, flowering time, meristem initiation or transition, floral organ development or response to stress. Gene expression profiling demonstrated that these genes had either similar or distinct expression pattern in wheat. Several genes regulating spike development were expressed in the early spike, of which Earliness per se 3 (Eps-3) was found might function in the initiation of spikelet meristem. Our study helps uncover important genes associated with apical meristem morphology and development in wheat.

Xiaoyan 6, one of the most important founder parents in wheat, possesses many superior agronomic traits and has played a crucial role in Chinese wheat breeding programs. In this study, a panel of 66 elite wheat accessions derived from Xiaoyan 6 was planted in four growing seasons; genome-wide association study (GWAS) was performed for six yield-related traits using the wheat 90K genotyping assay. A total of 803 significant marker-trait associations (MTAs) that explained up to 35.0% of the phenotypic variation were detected. Of these, the locus QTkw-5B which contains 19 MTAs for thousand kernel weight (TKW) was consistently detected in three growing seasons and confirmed in a recombinant inbred line (RIL) population by developing simple sequence repeats (SSR) and kompetitive allele-specific PCR (KASP) markers. The locus QPh-3A containing eight repetitive MTAs for plant height (PH) was consistently identified in all the four growing seasons and validated in a RIL population by developing SSR markers. The transmission of Xiaoyan 6 allele indicated that the favorite allele of QPh-3A was strongly selected in breeding programs. Comparing with previous studies, QTkw-5B and QPh-3A should be novel QTL. The locus QFss-2D for fertile spikelet number per spike (FSS) was identified and then validated in three bi-parental populations. This locus controlled various spike-related traits and may be a key spike polymorphic locus. This study could provide insight into dissecting yield-related traits in the breeding population and reliable molecular markers that might be valuable for marker-assisted selection in wheat high-yield breeding programs.

An allele of Pm2 for wheat powdery mildew resistance was identified in a putative Agropyron cristatum -derived line and used in wheat breeding programs. Powdery mildew (caused by Blumeria graminis f. sp. tritici, Bgt) is one of the most devastating wheat diseases worldwide. It is important to exploit varied sources of resistance from common wheat and its relatives in resistance breeding. KM2939, a Chinese breeding line, exhibits high resistance to powdery mildew at both the seedling and adult stages. It carries a single dominant powdery mildew resistance (Pm) allele of Pm2, designated Pm2b, the previous allelic designation Pm2 will be re-designated as Pm2a. Pm2b was mapped to chromosome arm 5DS and flanked by sequence characterized amplified region (SCAR) markers SCAR112 and SCAR203 with genetic distances of 0.5 and 1.3 cM, respectively. Sequence tagged site (STS) marker Mag6176 and simple sequence repeat (SSR) marker Cfd81 co-segregated with SCAR203. Pm2b differs in specificity from donors of Pm2a, Pm46 and PmLX66 on chromosome arm 5DS. Allelism tests indicated that Pm2b, Pm2a and PmLX66 are allelic. Therefore, Pm2b appears to be a new allele at the Pm2 locus. The closely linked markers were used to accelerate transfer of Pm2b to wheat cultivars in current production.

At present, most of released wheat cultivars or breeding lines in China are susceptible to powdery mildew (Pm) (caused by Blumeria graminis f. sp. tritici, Bgt), so there is an urgent need to rapidly transfer effective and broad-spectrum Pm resistance genes into elite cultivars/lines. Near-isogenic lines (NILs) with short target gene region are very important in molecular breeding and map-based cloning and can be developed by combining marker-assisted selection and conventional phenotypic identification. However, no Pm gene NILs were reported by using this method in the previous studies. A new broad-spectrum dominant resistance gene Pm2b, derived from the Chinese wheat breeding line KM2939, conferred high resistance to Pm at both the seedling and adult stages. In this study, with the aid of forward and background selection (FS and BS) using molecular markers, the Pm2b gene was introgressed into three elite susceptible commercial cultivars Shimai 15, Shixin 828, and Kenong 199 through the back-crossing procedure. With the appropriate backcrossing generations, selected population sizes and marker number for BS, the homozygous resistant BC3F2:3 NILs of Pm2b gene in the three genetic backgrounds with the highest recipient genome composition of about 99%, confirmed by simple sequence repeat markers and 660K single nucleotide polymorphic array, were developed and evaluated for the powdery mildew resistance and agronomic traits. The different resistance and similar or improved agronomic performance between Pm2b NILs and their corresponding recurrent parents indicated their potential value in the marker-assisted breeding of the Pm2b gene. Moreover, the development of four flanked diagnostic markers (CFD81, BWM25, BWM20, and BWM21) of the Pm2 gene can effectively assist the forward selection and accelerate the transfer and use of this resistance gene.

The improvement for drought tolerance requires understanding of the genetic control of wheat (Triticum aestivum L.) reaction to drought. In this study, a set of 131 recombinant inbred lines of wheat were investigated under well-watered (WW) and drought stress (DS) environments across 2 years to map quantitative trait loci (QTLs) for yield and physiological traits. A total of 225 QTLs were detected, including 32 non-environment-specific loci that were significant in both DS and WW, one drought-specific locus and two watering-specific loci. Three consistently-expressed QTLs (QTkw-3A.2, QTss-1A, and QScn-7A.1) were identified in at least three environments and the QTkw-1D.1 was significant in DS across the 2 years. By unconditional and conditional QTL analysis, spike number per plant and kernel number per spike were more important than thousand-kernel weight for grain yield (GY) at the given genetic background. Meta-analysis identified 67 meta-QTLs that contained QTLs for at least two traits. High frequency co-location of QTLs was found among either the spike-related traits or the six physiological traits. Four photosynthesis traits (CHL, LWUE, P N, and C i) were co-located with GY and/or yield components on various MQTLs. The results provided QTLs that warrant further study for drought tolerance breeding and are helpful for understanding the genetic basis of drought tolerance and the genetic contribution of yield components to GY at individual QTL level in wheat.

Identification and mapping new powdery mildew resistance (Pm) genes is important for resistance breeding in wheat. Common wheat (Triticum aestivum L.) line X3986-2 was tested against 27 isolates of Blumeria graminis f. sp. tritici. To identify the Pm gene(s) in X3986-2, an F2 population and its derived F2:3 lines were developed from a cross between X3986-2 and susceptible line Mingxian169. Segregation ratios indicated the presence of a single dominant Pm locus, tentatively designated PmX3986-2. Bulked segregant analysis was applied to screen for molecular markers linked to PmX3986-2. Two sequence characterized amplified region (SCAR) markers SCAR112 and SCAR203, and five simple sequence repeat markers CFD40, CFD78, CFD81, GWM293 and WMC443 on chromosome 5D were linked to PmX3986-2, with CFD81 and SCAR112 flanking PmX3986-2 at 0.6 and 1.5 cM, respectively. This suggests that PmX3986-2 may be a novel allele of loci Pm2, Pm46 and PmLX66 on chromosome arm 5DS. PmX3986-2 with its tightly linked DNA markers should be useful for broadening the genetic basis of Pm and rapidly transferring the resistance gene to susceptible cultivars or for us in gene pyramiding for resistance breeding.

Stripe rust resistance gene YrC591, present in wheat cultivar C591, is effective against currently important Puccinia striiformis Westend. f. sp. tritici isolates in China. An F2:3 population (127 lines) was developed by crossing C591 with susceptible cultivar Taichung 29. Thirty four simple sequence repeat (SSR) and 155 sequence tagged site (STS) markers located on chromosome 7BL were used to perform bulk segregant analysis. Eight SSR markers, cfa2040, wmc273, wmc166, gwm984, barc32 wmc276, barc182 and gwm146, and 6 STS markers, mag1714, mag1757, mag1811, BE425120, BE471173 and BG607810, were polymorphic between the parents and contrasting resistant and susceptible DNA pools. F2:3 lines were genotyped with these polymorphic markers. Linkage analysis indicated that YrC591 was flanked by Xmag1714 and Xbarc182 with genetic distances of 1.2 and 0.4 cM, respectively. In addition, validation of the SSR markers cfa2040, wmc273 and barc32, and STS markers mag1714 and BE425120 was carried out using wheat lines with C591 as a parent, indicating that these markers should be effective in tracing this gene in marker-assisted selection.

The present study identified some new important genomic regions and demonstrated the availability of conditional analysis in dissecting QTLs induced by environmental factors. The high input and low use efficiency of nutrient fertilizers require knowledge of the genetic control of crop reaction to nutrient supplements. In this study, 14 morphological and 8 physiological traits of a set of 182 wheat (Triticum aestivum L.) recombinant inbred lines (Xiaoyan 54 × Jing 411) were investigated in six environments to map quantitative trait loci (QTLs). The influence of nitrogen (N) and phosphorus (P) fertilization on QTL expression was studied by unconditional and conditional analysis. A total of 117 and 30 QTLs were detected by unconditional and conditional analysis, respectively, among which 21 were common for both methods. Thirty-four QTL clusters were identified. Eighteen conserved QTLs (15.4 % of the 117 QTLs) between years, but within nutritional treatment were found. The three major QTLs on chromosomes 2D, 4B and 6A were coincident with Rht8, Rht-B1b and TaGW2, respectively. The other two important intervals on chromosomes 4B and 7A for yield component traits were newly detected QTLs that warrant further study. By conditional analysis, spikelet number per spike was found to be induced by P fertilization mostly, whereas N fertilization had more effects on the expression of the QTLs for nitrogen concentration and utilization efficiency traits. QTLs that respond to N and P interactions were also detected. The results are helpful for understanding the genetic basis of N utilization efficiency in wheat under different N and P supplement environments and provide evidence for the availability of conditional analysis in dissecting QTLs induced by environmental factors.

Quantitative trait loci (QTLs) for salt tolerance with additive, epistatic and QTL × treatment interaction effects at seedling stage in wheat were identified. A set of 131 recombinant inbred lines derived from cross Chuan 35050 × Shannong 483 were evaluated under salt stress and normal conditions. Wide variation was found for all studied traits. A total of 18 additive and 16 epistatic QTLs were detected, among which five and 11 were with significant QTL × treatment effects. Ten QTL clusters were identified, and each may represent a single gene or closely linked genes. The locus controlling shoot K+/Na+ concentration ratio and shoot Na+ concentration on chromosome 5A may be identical to Nax2. The interval Xgwm6-Xgwm538 on chromosome 4B for total dry weight was also identified in a previous study, both near the marker Xgwm6. The marker Xgwm6 may be useful for marker-assisted selection. Six pairs of homoeologous QTLs were detected, showing synteny among the A, B and D genomes. These results facilitate understanding the mechanisms and the genetic basis of salt tolerance in wheat.

Powdery mildew has a negative impact on wheat production. Novel host resistance increases the diversity of resistance genes and helps to control the disease. In this study, wheat line FG-1 imported from France showed a high level of powdery mildew resistance at both the seedling and adult stages. An F2 population and F2:3 families from the cross FG-1 × Mingxian 169 both fit Mendelian ratios for a single dominant resistance gene when tested against multiple avirulent Blumeria tritici f. sp. tritici (Bgt) races. This gene was temporarily designated PmFG. PmFG was mapped on the multi-allelic Pm2 locus of chromosome 5DS using seven SSR, 10 single nucleotide polymorphism (SNP)-derived and two SCAR markers with the flanking markers Xbwm21/Xcfd81/Xscar112 (distal) and Xbwm25 (proximal) at 0.3 and 0.5 cM being the closest. Marker SCAR203 co-segregated with PmFG. Allelism tests between PmFG and documented Pm2 alleles confirmed that PmFG was allelic with Pm2. Line FG-1 produced a significantly different reaction pattern compared to other lines with genes at or near Pm2 when tested against 49 Bgt isolates. The PmFG-linked marker alleles detected by the SNP-derived markers revealed significant variation between FG-1 and other lines with genes at or near Pm2. It was concluded that PmFG is a new allele at the Pm2 locus. Data from seven closely linked markers tested on 31 wheat cultivars indicated opportunities for marker-assisted pyramiding of this gene with other genes for powdery mildew resistance and additional traits.

Low concentration or deficiencies of bioavailable micronutrients in staple food result in micronutrient malnutrition. The understanding of the genetic basis for micronutrient concentration in wheat grain can provide useful information for wheat biofortification breeding by marker-assisted selection (MAS). In the present study, grain zinc (Zn), iron (Fe) and protein concentrations were analyzed in a recombinant inbred line (RIL) population under two environments to detect quantitative trait loci (QTLs) and their interactions. Nine additive and four epistatic QTLs were identified, among which six and four, respectively, were effective at the two environments. Three intervals that affected two or three traits were found on chromosomes 4B and 5A, which indicated a common genetic basis among grain Zn, Fe and protein concentrations. Multi-trait composite interval mapping analysis detected five intervals that influenced more than one trait, among which three were identified at both the environments and two could affect all of the three traits. The interval on chromosome 5A for grain Fe and protein concentrations was co-located with QTLs controlling the same traits in a previous study, both near the marker Xgwm154. The molecular markers closely linked to the QTLs, e.g., Xgwm154, may be useful in wheat biofortification breeding by MAS. The interval Xcfd21641-NP21.1 on chromosome 6A which was effective for all the three traits may be an ortholog of Gpc-B1.

Since it was commercialized in 2008, Liangxing 66 is one of the most widely grown cultivars of wheat (Triticum aestivum L.) in winter and facultative wheat-producing regions in northern China. This cultivar displays broad-spectrum resistance to isolates of powdery mildew. To identify the powdery mildew resistance gene in Liangxing 66, genetic analysis and molecular mapping were conducted using the F2 populations and F2:3 families derived from the reciprocal crosses of Liangxing 66 and the susceptible cultivar Jingshuang 16. A single dominant gene, tentatively designated PmLX66, conferred resistance in Liangxing 66 to the powdery mildew isolate E09. The results of molecular mapping indicated that this gene was located on the short arm of chromosome 5D and flanked by SCAR203 and Xcfd81 at genetic distances of 0.4 and 2.8 cM, respectively, which is similar to the position of locus Pm2. However, PmLX66 and Pm2 showed different reactions to five of the 42 isolates of powdery mildew tested. Together, these results indicated that PmLX66 was most likely an allele of Pm2. Based on its superior yield and agronomic performance, in combination with powdery mildew resistance, Liangxing 66 is useful as a promising parent for control of powdery mildew and for the development of new disease-resistant cultivars.

Quantitative trait loci (QTLs) with additive (a), additive × additive (aa) epistatic effects, and their treatmental interactions (at and aat) were studied under salt stress and normal conditions at seedling stage of wheat (Triticum aestivum L.). A set of 182 recombinant inbred lines (RILs) derived from cross Xiaoyan 54 × Jing 411 were used. A total of 29 additive QTLs and 17 epistasis were detected for 12 traits examined, among which eight and seven, respectively, were identified to have QTL × treatment effects. Physiological traits rather than biomass traits were more likely to be involved in QTL × treatment interactions. Ten intervals on chromosomes 1A, 1D, 2A (two), 2D, 3B, 4B, 5A, 5B and 7D showed overlapping QTLs for different traits; some of them represent a single locus affecting different traits and/or the same trait under both treatments. Eleven pairs of QTLs were detected on seemingly homoeologous positions of six chromosome groups of wheat, showing synteny among the A, B and D genomes. Ten pairs were detected in which each pair was contributed by the same parent, indicating a strong genetic plasticity of the QTLs. The results are helpful for understanding the genetic basis of salt tolerance in wheat and provide useful information for genetic improvement of salt tolerance in wheat by marker-assisted selection.

To develop a set of molecular markers specific for the chromosome arms of rye, a total of 1,098 and 93 primer pairs derived from the expressed sequence tag (EST) sequences distributed on all 21 wheat chromosomes and 7 rye chromosomes, respectively, were initially screened on common wheat ‘Chinese Spring’ and rye cultivar ‘Imperial’. Four hundred and fourteen EST-based markers were specific for the rye genome. Seven disomic chromosome addition lines, 10 telosomic addition lines and 1 translocation line of ‘Chinese Spring-Imperial’ were confirmed by genomic in situ hybridization and fluorescencein situ hybridization, and used to screen the rye-specific markers. Thirty-one of the 414 markers produced stable specific amplicons in ‘Imperial’, as well as individual addition lines and were assigned to 13 chromosome arms of rye except for 6RS. Six rye cultivars, wheat cultivar ‘Xiaoyan 6’ and accessions of 4 wheat relatives were then used to test the specificity of the 31 EST-based markers. To confirm the specificity, 4 wheat-rye derivatives of ‘Xiaoyan 6 × German White’, with chromosomes 1RS, 2R and 4R, were amplified by some of the EST-based markers. The results indicated that they can effectively be used to detect corresponding rye chromosomes or chromosome arms introgressed into a wheat background, and hence to accelerate the utilization of rye genes in wheat breeding.

Molecular identification of wheat powdery mildew resistance (Pm) genes in breeding lines or cultivars is important for resistance breeding. Wanfengjian 34, a Chinese wheat breeding line with desirable agricultural performance in China, showed high level of resistance to powdery mildew at both seedling and adult stages. Genetic analysis demonstrated that the resistance to different Blumeria graminis f. sp. tritici isolates in Wanfengjian 34 was controlled by a single dominant Pm gene, temporarily designated PmWFJ. PmWFJ was mapped to chromosome arm 5DS and flanked by the simple sequence repeat marker Cfd81 and sequence characterized amplified region marker SCAR112 with genetic distance of 0.1 and 1.0 cM, respectively. PmWFJ shared the same locus with the documented Pm genes on the multi-allelic Pm2 locus by allelism tests between PmWFJ and Pm2 alleles, but showed unique powdery mildew reaction pattern compared with these documented Pm genes. Therefore, PmWFJ appears to be a new allele at the documented Pm2 locus. Four closely linked markers of PmWFJ were validated the applicability in 131 cultivar backgrounds for marker-assisted selection.

Nutrient use efficiency (NuUE), comprising nutrient uptake and utilization efficiency, is regarded as one of the most important factors for wheat yield. In the present study, six morphological, nine nutrient content and nine nutrient utilization efficiency traits were investigated at the seedling stage using a set of recombinant inbred lines (RILs), under hydroponic culture of 12 treatments including single nutrient levels and two- and three-nutrient combinations treatments of N, P and K. For the 12 designed treatments, a total of 380 quantitative trait loci (QTLs) on 20 chromosomes for the 24 traits were detected. Of these, 87, 149 and 144 QTLs for morphological, nutrient content and nutrient utilization efficiency traits were found, respectively. Using the data of the average value (AV) across 12 treatments, 70 QTLs were detected for 23 traits. Most QTLs were located in new marker regions. Twenty-six important QTL clusters were mapped on 13 chromosomes, 1A, 1B, 1D, 2B, 3A, 3B, 4A, 4B, 5D, 6A, 6B, 7A and 7B. Of these, ten clusters involved 147 QTLs (38.7%) for investigated traits, indicating that these 10 loci were more important for the NuUE of N, P and K. We found evidence for cooperative uptake and utilization (CUU) of N, P and K in the early growth period at both the phenotype and QTL level. The correlation coefficients (r) between nutrient content and nutrient utilization efficiency traits for N, P and K were almost all significantly positive correlations. A total of 32 cooperative CUU loci (L1-L32) were found, which included 190 out of the 293 QTLs (64.8%) for the nutrient uptake and utilization efficiency traits, indicating that the CUU-QTLs were common for N, P and K. The CUU-QTLs in L3, L7, L16 and L28 were relatively stable. The CUU-QTLs may explain the CUU phenotype at the QTL level.

Molecular markers are important tools that have been used to identify the short arm of rye chromosome 1R (1RS) which contains many useful genes introgressed into wheat background. Wheat expressed sequence tag (EST) sequences are valuable for developing molecular markers since ESTs are derived from gene transcripts and more likely to be conserved between wheat and its relative species. In the present study, 35 sequence-tagged site (STS) primers were designed based on EST sequences distributed on homology group 1 chromosomes of Triticum aestivum and used to screen specific markers for chromosome 1RS of Secale cereale . Two primer pairs different from the early studies, STS WE3 , which amplified a 1680-bp and a 1750-bp fragment, and STS WE126 , which produced a 850-bp fragment from rye genome, were proved to be specific to chromosome 1RS since the corresponding fragments were only amplified from 1R chromosome addition line and wheat-rye lines with chromosome 1RS, but not from wheat-rye 2R-7R chromosome addit...

The grain yield of normal oil maize ( Zea mays L.) might increase when pollinated by high oil maize (HOM) hybrids because of heterosis. To testify that the grain yield increase might be a result of improved photosynthetic rate and related traits, the normal oil maize (NOM) hybrid, Nongda108, was cross-pollinated by three HOM hybrids, HOM202, HOM115 and HOM4515 (for short as ND108pHOM202 , ND108pHOM115 and ND108pHOM4515 ). We found that the ND108pHOM202 and ND108pHOM115 exhibited higher net photosynthetic rate ( P n ), accompanied by larger stomatal conductance ( g s ) and transpiration rate ( E ). Moreover, delayed leaf senescence was observed in their leaves, including larger leaf area index (LAI) and higher Chl content and Chl a / b ratio. Apart from higher phosphoenolpyruvate carboxylase (PEPCase) activity, the soluble proteins were also higher in the two cross-pollinations. The higher leaf photosynthesis could explain the grain increase in ND108pHOM202 and ND108pHOM115 . However, ND108pHOM4515 exhibited a decreased photosynthetic characteristic and yield performance. Significantly positive relation between grain yield and biomass ( r 2 = 0.96, P P n and biomass ( r 2 = 0.74, P

It is important to detect wheat broad-spectrum powdery mildew resistance (Pm) genes in commercial cultivars with superior agronomic performance. YingBo 700, a Chinese commercial cultivar registered and released in 2012, showed a broad spectrum of resistance to 48 of 49 tested Blumeria graminis f. sp tritici (Bgt) isolates from different regions of China. Genetic analysis of F2 populations from the crosses YingBo 700 × Mingxian 169 and YingBo 700 × Shimai 15 and F2:3 lines from YingBo 700 × Mingxian 169 demonstrated that the resistance was controlled by a single dominant resistance gene, which was temporarily designated PmYB. Using molecular markers and bulked segregant analysis of the F 2:3 lines from YingBo 700 × Mingxian 169, PmYB was mapped to the multi-allelic Pm2 region of chromosome arm 5DS co-segregated with the sequence-characterized amplified region (SCAR) marker SCAR112 and flanked by the simple sequence repeat (SSR) marker Cfd81 and SCAR marker SCAR203 with genetic distance of 0.9 and 1.9 cM, respectively. YingBo 700 showed a unique response pattern to different Bgt isolates compared with the lines with documented Pm2a, Pm2b, Pm48, PmLX66, PmW14, PmZ155 and PmX3986-2 on chromosome arm 5DS. Therefore, PmYB appears to be a novel allele in this multi-allelic region. In order to validate applicability of the closely linked markers for marker-assisted selection, the closely linked markers Cfd81, SCAR112, SCAR203 and Mag6176 were evaluated for applicability in diagnosing the resistance gene in 62 cultivars from different regions of China. The results will contribute to rapidly transfer of PmYB into more cultivars.

A new broad-spectrum powdery mildew resistance allele Pm2c was identified and mapped in Chinese wheat landrace Niaomai. Chinese wheat landrace Niaomai showed resistance to 27 of 28 Chinese Blumeria graminis f. sp tritici (Bgt) races. Genetic analysis of an F2 population and its derived F2:3 families from the cross Niaomai × Mingxian 169 and backcross population, Niaomai/2*Mingxian 169, indicated that the resistance of Niaomai to Bgt races was conferred by a single dominant resistance gene, temporarily designated PmNM. Molecular tagging showed that PmNM was located on chromosome 5DS and flanked by SSR markers Xcfd81 and Xcfd78 with the genetic distances of 0.1/0.4 cM and 4.9/7.5 cM, respectively. Niaomai showed a different array of responses compared to lines with Pm2a, Pm2b, PmD57-5D, PmLX66, PmX3986-2 and Pm48 genes, sharing the same Xcfd81 allele but differing from Xcfd78 allele for Pm2a and Pm2b lines. Allelism tests based on crosses of Niaomai with Ulka/8*Cc and KM2939 showed that PmNM is allelic to Pm2a and Pm2b. We concluded that PmNM is a new allele of Pm2, re-designated Pm2c. Pm2c could be transferred into wheat cultivars by marker-assisted selection to improve the powdery mildew resistance of breeding cultivars/lines.

Using the nullisomic back-cross procedure, four wheat-rye chromosome substitution 2R (2D) lines with different agronomic performance, designated WR02-145-1, WR01-145-2, WR02-145-3, and WR02-145-4, were produced from a cross between 2D nullisomic wheat (Triticum aestivum L. cv. “Xiaoyan 6”) and rye (Secale cereale L. cv. “German White”). The chromosomal constitution of 2n=42=21 in WR02-145 lines was confirmed by cytological and molecular cytogenetic methods. Using genomic in situ hybridization on root tip chromosome preparations, a pair of intact rye chromosomes was detected in the WR02-145 lines. PCR using chromosome-specific primers confirmed the presence of 2R chromosomes of rye in these wheat-rye lines, indicating that WR02-145 lines are disomic chromosome substitution lines 2R(2D). The WR02-145 lines are resistant to the powdery mildew (Erysiphe graminis DC. f. sp. tritici E. Marchal) isolates prevalent in northern China and may possess gene(s) for resistance to powdery mildew, which differ from the previously identified Pm7 gene located on chromosome 2RL. The newly developed “Xiaoyan 6”- “German White” 2R (2D) chromosome substitution lines are genetically stable, show desirable agronomic traits, and are expected to be useful in wheat improvement. (Managing editor: Li-Hui Zhao)

The objective of this study was to map QTLs for N uptake (NUP) in wheat, and to investigate factors influencing NUP. Two independent field trials with low N (LN) and high N (HN) treatments were conducted in the growing seasons of 2002–2003 (trial 1) and 2003–2004 (trial 2) to measure NUP per plant (N accumulated in the aerial part at maturity stage) of a doubled haploid (DH) population consisting of 120 DH lines derived from winter wheat varieties Hanxuan 10 and Lumai 14. A hydroponic culture with all nutrients supplied sufficiently was conducted to investigate shoot dry weight (SDW), root dry weight (RDW), tiller number (TN) and NUP (total plant N uptake) per plant of this mapping population at seedling stage. SDW, RDW, TN and NUP investigated in the hydroponic culture were significantly and positively correlated with each other, and with NUP under both LN and HN conditions in the field trials. Nine and eight QTLs for NUP were detected under LN and HN conditions in the field trials, respectively. Four to five QTLs for SDW, RDW, TN and NUP were detected in the hydroponic culture. One SDW QTL, three RDW QTLs, two TN QTLs detected in the hydroponic culture were linked with QTLs for NUP under LN or HN condition in the field trials. The positive correlation and genetic linkage for the traits between the field trials and the hydroponic culture demonstrated that greater seedling vigor of root and shoot is an important factor influencing N uptake in wheat.

Rye is an important and valuable gene resource for wheat improvement. However, due to extensive growing of cultivars with disease resistance genes from short arm of rye chromosome 1R and coevolution of pathogen virulence and host resistance, these cultivars successively lost resistance to pathogens. Identification and deployment of new resistance gene sources in rye are, therefore, of especial importance and urgency. A new wheat–rye line, designated as WR41-1, was produced through distant hybridization and chromosome engineering protocols between common wheat cultivar Xiaoyan 6 and rye cultivar German White. It was proved to be a new wheat–rye T4BL·4RL and T7AS·4RS translocation line using sequential genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization (mc-FISH), and expressed sequence tag-simple sequence repeat (EST-SSR) marker analysis. WR41-1 showed high levels of resistance to powdery mildew (Blumeria graminis f. sp. tritici, Bgt) pathogens prevalent in China at the adult growth stage and 13 of 23 Bgt isolates tested at the seedling stage. According to its resistant pattern to 23 different Bgt isolates, WR41-1 may possess new gene(s) for resistance to powdery mildew, which differed from previously identified and known powdery mildew genes from rye (Pm7, Pm8, Pm17, and Pm20). In addition, WR41-1 was cytologically stable, had a desirable fertility, and is expected to be useful in wheat improvement.