Wheat Cultivar Chinese Spring Research Articles

Transcriptome analysis of Lr19-virulent mutants provides clues for the AvrLr19 of Puccinia triticina.

Wheat leaf rust caused by Puccinia triticina (Pt) remains one of the most destructive diseases of common wheat worldwide. Understanding the pathogenicity mechanisms of Pt is important to control wheat leaf rust. The urediniospores of Pt race PHNT (wheat leaf rust resistance gene Lr19-avirulent isolate) were mutagenized with ethyl methanesulfonate (EMS), and two Lr19-virulent mutants named M1 and M2 were isolated. RNA sequencing was performed on samples collected from wheat cultivars Chinese Spring and TcLr19 infected with wild-type (WT) PHNT, M1, and M2 isolates at 14 days post-inoculation (dpi), respectively. Screening AvrLr19 candidates by quantitative reverse transcription PCR (qPCR) and Agrobacterium-mediated transient assays in Nicotiana benthamiana. 560 genes with single nucleotide polymorphisms (SNPs) and insertions or deletions (Indels) from non-differentially expressed genes were identified. Among them, 10 secreted proteins were screened based on their fragments per kilobase of exon model per million mapped reads (FPKM) values in the database. qPCR results showed that the expression profiles of 7 secreted proteins including PTTG_27471, PTTG_12441, PTTG_28324, PTTG_26499, PTTG_06910, PTTG_26516, and PTTG_03570 among 10 secreted proteins in mutants were significantly different with that in wild-type isolate after infection wheat TcLr19 and might be related to the recognition between Lr19 and AvrLr19. In addition, a total of 216 differentially expressed genes (DEGs) were obtained from three different sample comparisons including M1-vs-WT, M2-vs-WT, and M1-vs-M2. Among 216 DEGs, 15 were predicted to be secreted proteins. One secreted protein named PTTG_04779 could inhibit programmed progress of cell death (PCD) induced by apoptosis-controlling genes B-cell lymphoma-2 associated X protein (BAX) on Nicotiana benthamiana, indicating that it might play a virulence function in plant. Taken together, total 8 secreted proteins, PTTG_04779, PTTG_27471, PTTG_12441, PTTG_28324, PTTG_26499, PTTG_06910, PTTG_26516, PTTG_03570 are identified as AvrLr19 candidates. Our results showed that a large number of genes participate in the interaction between Pt and TcLr19, which will provide valuable resources for the identification of AvrLr19 candidates and pathogenesis-related genes.

TaPR1 Interacts With TaTLP1 via the αIV Helix to Be Involved in Wheat Defense to Puccinia triticina Through the CAPE1 Motif.

Pathogenesis-related (PR) proteins play important roles in plant defense response and systemic acquired resistance (SAR). PR1 has antifungal activity against many plant pathogens. In our previous study, RNA sequencing (RNA-seq) was conducted on resistant wheat line TcLr19 and sensitive wheat cultivar Chinese Spring inoculated with Puccinia triticina (Pt) race PHNT. In this study, seven salicylic acid (SA)-induced TaPR1 genes involved in plant disease resistance were found in the RNA-seq library. Quantitative PCR (qPCR) results showed that TaPR1-4 was most induced by Pt among these seven TaPR1 genes in the incompatible interaction. Yeast two-hybrid (Y2H) results showed that TaPR1-4 interacted with TaTLP1 via the αIV helix. Protein-mediated phenotyping assays in vivo and antifungal activity in vitro demonstrated that wheat leaves infiltrated with pure TaPR1-4 protein developed significantly less disease compared to control leaves. This effect was correlated with a strong increase in defense gene expression, and resistance activity was dependent on the CAPE1 motif located in the C-terminal region of TaPR1-4. These findings increase current knowledge regarding the interaction of TaPR1 and TaTLP1 and provide new insights on the role of TaPR1 protein in the resistance of wheat to Pt.

Distant somatic hybridization alters the structure of wheat root bacterial microbiota

AbstractRoot microbiota affect the host plant's adaptation to adverse environments. The structure and function of plant root bacterial communities are affected by soil physicochemical properties and plant genotypes. Distant hybridization is a common breeding strategy to introgress the excellent genes of wild relatives into the crop genome. The effect of distant hybridization on the crop root microbiota has not been reported. In this study, the structure and function of root bacterial community of the somatic hybrid wheat introgression lines SR3 and SR4, which have been respectively reported to be tolerant to salt and saline‐alkali stress, and their parent wheat cultivar JN177 as well as common wheat cultivar Chinese Spring (CS) were analyzed using 16S rRNA high‐throughput sequencing technology. The results showed that distant hybridization altered the structure and function of wheat (Triticum aestivum L.) root bacterial community. Compared with the parent JN177, some bacterial genera related to plant growth and stress resistance showed higher abundance in SR3 and SR4. Intriguingly, compared with parent JN177, the root bacterial communities of SR3 and SR4 are more similar to that of CS. This indicated that the process of distant hybridization may alter key genes regulating wheat root microbiota.

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

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

Transfer of fusarium head blight resistance from Thinopyrum elongatum to bread wheat cultivar Chinese Spring

The diploid form of tall wheatgrass, Thinopyrum elongatum (Host) D.R. Dewey (2n = 2x = 14, EE genome), has a high level of resistance to fusarium head blight. The symptoms did not spread beyond the inoculated florets following point inoculation. Using a series of E-genome chromosome additions in a bread wheat cultivar Chinese Spring (CS) background, the resistance was found to be localized to the long arm of chromosome 7E. The CS mutant ph1b was used to induce recombination between chromosome 7E, present in the 7E(7D) substitution and homoeologous wheat chromosomes. Multivalent chromosome associations were detected in the BC1 hybrids, confirming the effectiveness of the ph1b mutant. Genetic markers specific for chromosome 7E were used to estimate the size of the 7E introgression in the wheat genome. Using single sequence repeat (SSR) markers specific for homoeologous wheat chromosome 7, introgressions were detected on wheat chromosomes 7A, 7B, and 7D. Some of the introgression lines were resistant to fusarium head blight.

Gene structure of three kinds of vacuolar-type Na+/H+ antiporters including TaNHX2 transcribed in bread wheat.

The vacuolar-type sodium/proton antiporter is considered to play an important role in withstanding salt stress by transporting sodium ions into vacuoles. In this study, the gene structures of three kinds of vacuolar-type antiporters transcribed in bread wheat under salt stress were analyzed. After spraying 0.5 M NaCl to seedlings of wheat cultivar Chinese Spring, 1,392~1,400 bp cDNA fragments were isolated by RT-PCR using primers designed from common regions in rice OsNHX1 and Atriplex subcordata AgNHX1. Next, the entire structure of the genomic DNA and cDNA were determined via CapFishing-5’ Rapid Amplification of cDNA Ends (RACE), 3’RACE, and genomic PCR cloning. As a result, 3 kinds of vacuolar-type Na+/H+ antiporter genes, TaNHXa (genome DNA 4,255 bp, cDNA 2,414 bp, 539 a.a.), TaNHXb (gDNA 4,167 bp, cDNA 1,898 bp, 539 a.a.) and TaNHXc (gDNA 4,966 bp, cDNA 1,928 bp, 547 a.a.), were identified. They encode 12 transmembrane domains containing third domain’s amyloid binding sites (FFIYLLPP), characteristic of the vacuolar-type Na+/H+ antiporter, binding to the cell vacuolar membrane. TaNHXa, b and c consisting of 14 exons and 13 introns were 22~55 % longer than A. thaliana AtNHX1 in total length. TaNHXa (TaNHX2) showed homogeneity with OsNHX1, while TaNHXb and c were phylogenetically independent.

Identification of a novel ERF gene, TaERF8, associated with plant height and yield in wheat

BackgroundEthylene Responsive Factor (ERF) is involved in various processes of plant development and stress responses. In wheat, several ERFs have been identified and their roles in mediating biotic or abiotic stresses have been elucidated. However, their effects on wheat plant architecture and yield-related traits remain poorly studied.ResultsIn this study, TaERF8, a new member of the ERF family, was isolated in wheat (Triticum aestivum L.). Three homoeologous TaERF8 genes, TaERF8-2A, TaERF8-2B and TaERF8-2D (named according to sub-genomic origin), were cloned from the common wheat cultivar Chinese Spring. The three homoeologs showed highly similar protein sequences, with identical AP2 domain. Whereas homoeologs sequence polymorphism analysis allowed the establishment of ten, two and three haplotypes, respectively. Expression analysis revealed that TaERF8s were constitutively expressed through entire wheat developmental stages. Analysis of related agronomic traits of TaERF8-2B overexpressing transgenic lines showed that TaERF8-2B plays a role in regulating plant architecture and yield-related traits. Association analysis between TaERF8-2B haplotypes (Hap-2B-1 and Hap-2B-2) and agronomic traits showed that TaERF8-2B was associated with plant height, heading date and 1000 kernel weight (TKW). The TaERF8-2B haplotypes distribution analysis revealed that Hap-2B-2 frequency increased in domesticated emmer wheat and modern varieties, being predominant in five major China wheat producing zones.ConclusionThese results indicated that TaERF8s are differentially involved in the regulation of wheat growth and development. Haplotype Hap-2B-2 was favored during domestication and in Chinese wheat breeding. Unveiling that the here described molecular marker TaERF8-2B-InDel could be used for marker-assisted selection, plant architecture and TKW improvement in wheat breeding.

Machine learning analyses of methylation profiles uncovers tissue‐specific gene expression patterns in wheat

DNA methylation is a mechanism of epigenetic modification in eukaryotic organisms. Generally, methylation within genes promoter inhibits regulatory protein binding and represses transcription, whereas gene body methylation is associated with actively transcribed genes. However, it remains unclear whether there is interaction between methylation levels across genic regions and which site has the biggest impact on gene regulation. We investigated and used the methylation patterns of the bread wheat cultivar Chinese Spring to uncover differentially expressed genes (DEGs) between roots and leaves, using six machine learning algorithms and a deep neural network. As anticipated, genes with higher expression in leaves were mainly involved in photosynthesis and pigment biosynthesis processes whereas genes that were not differentially expressed between roots and leaves were involved in protein processes and membrane structures. Methylation occurred preponderantly (60%) in the CG context, whereas 35 and 5% of methylation occurred in CHG and CHH contexts, respectively. Methylation levels were highly correlated (r=0.7 to 0.9) between all genic regions, except within the promoter (r=0.4 to 0.5). Machine learning models gave a high (0.81) prediction accuracy of DEGs. There was a strong correlation (p-value=9.20×10-10 ) between all features and gene expression, suggesting that methylation across all genic regions contribute to gene regulation. However, the methylation of the promoter, the CDS and the exon in CG context was the most impactful. Our study provides more insights into the interplay between DNA methylation and gene expression and paves the way for identifying tissue-specific genes using methylation profiles.

The NLR-Annotator Tool Enables Annotation of the Intracellular Immune Receptor Repertoire.

Disease resistance genes encoding nucleotide-binding and leucine-rich repeat (NLR) intracellular immune receptor proteins detect pathogens by the presence of pathogen effectors. Plant genomes typically contain hundreds of NLR-encoding genes. The availability of the hexaploid wheat (Triticum aestivum) cultivar Chinese Spring reference genome allows a detailed study of its NLR complement. However, low NLR expression and high intrafamily sequence homology hinder their accurate annotation. Here, we developed NLR-Annotator, a software tool for in silico NLR identification independent of transcript support. Although developed for wheat, we demonstrate the universal applicability of NLR-Annotator across diverse plant taxa. We applied our tool to wheat and combined it with a transcript-validated subset of genes from the reference gene annotation to characterize the structure, phylogeny, and expression profile of the NLR gene family. We detected 3,400 full-length NLR loci, of which 1,560 were confirmed as expressed genes with intact open reading frames. NLRs with integrated domains mostly group in specific subclades. Members of another subclade predominantly locate in close physical proximity to NLRs carrying integrated domains, suggesting a paired helper function. Most NLRs (88%) display low basal expression (in the lower 10 percentile of transcripts). In young leaves subjected to biotic stress, we found up-regulation of 266 of the NLRs To illustrate the utility of our tool for the positional cloning of resistance genes, we estimated the number of NLR genes within the intervals of mapped rust resistance genes. Our study will support the identification of functional resistance genes in wheat to accelerate the breeding and engineering of disease-resistant varieties.

At Least Five Major Genes Are Involved in the Avirulence of an Eleusine Isolate of Pyricularia oryzae on Common Wheat.

Pyricularia oryzae is composed of pathotypes that show host specificity at the plant genus level. To elucidate the genetic mechanisms of the incompatibility between the Eleusine pathotype (pathogenic on finger millet) and common wheat, an Eleusine isolate (MZ5-1-6) was crossed with a Triticum isolate (Br48) pathogenic on wheat, and resulting F1 cultures were sprayed onto common wheat cultivars Hope, Norin 4 (N4), and Chinese Spring (CS). On Hope, avirulent and virulent cultures segregated in a 3:1 ratio, suggesting that two avirulence genes are involved. They were tentatively designated as eA1 and eA2. On N4 and CS, the segregation ratio was not significantly deviated from the 7:1, 15:1, or 31:1 ratios, suggesting that three or more genes are involved. A comparative analysis of the segregation patterns suggested that two of these genes were eA1 and eA2. A complementation test indicated that the third gene (tentatively designated as eA3) was the Ao9 type of the PWT3 gene controlling the avirulence of Avena and Lolium isolates on wheat. The fourth gene (tentatively designated as eA4) was detected by backcrossing 200R72, an F1 culture lacking eA1, eA2, and eA3, with Br48. Comparative analyses of phenotypes and the presence and/or absence of molecular markers in the F1 population revealed that some cultures were avirulent on N4/CS in spite of lacking eA1, eA2, eA3, and eA4, indicating the presence of the fifth gene (tentatively designated as eA5). Taken together, we conclude that at least five avirulence genes are involved in the incompatibility between MZ5-1-6 and N4/CS.

Overexpression of the Wheat (Triticum aestivum L.) TaPEPKR2 Gene Enhances Heat and Dehydration Tolerance in Both Wheat and Arabidopsis.

Wheat (Triticum aestivum L.) yield and quality are adversely affected by heat, drought, or the combination of these two stresses in many regions of the world. A phosphoenolpyruvate carboxylase kinase-related kinase gene, TaPEPKR2, was identified from our previous heat stress-responsive transcriptome analysis of heat susceptible and tolerant wheat cultivars. Based on the wheat cultivar Chinese Spring genome sequence, TaPEPKR2 was mapped to chromosome 5B. Expression analysis revealed that TaPEPKR2 was induced by heat and polyethylene glycol treatment. To analyze the function of TaPEPKR2 in wheat, we transformed it into the wheat cultivar Liaochun10, and observed that the transgenic lines exhibited enhanced heat and dehydration stress tolerance. To examine whether TaPEPKR2 exhibits the same function in dicotyledonous plants, we transformed it into Arabidopsis, and found that its overexpression functionally enhanced tolerance to heat and dehydration stresses. Our results imply that TaPEPKR2 plays an important role in both heat and dehydration stress tolerance, and could be utilized as a candidate gene in transgenic breeding.

Genotypic effects on sugar and by-products of liquid hydrolysates and on saccharification of acid-insoluble residues from wheat straw.

Wheat straw is one of the major attractive resources for low-cost raw materials for renewable energy, biofuels and biochemicals. However, like other sources of lignocellulosic biomass, straw is a heterogeneous material due to its mixed origin from different tissue and cell types. Here, to examine the genotypic effects on biorefinery usage of wheat straw, straw obtained from different wheat cultivars and experimental lines was pretreated with dilute acid. Significant differences between cultivars were observed in the concentrations of glucose and toxic by-products of the liquid hydrolysates. A higher content of xylose than glucose was found in liquid hydrolysates from wheat straw, and the xylose content appeared to be affected by both environmental and genetic factors. Analysis using chromosome substitution lines of the common wheat cultivar Chinese Spring showed that chromosomes 2A and 3A from other wheat cultivars, Hope and Timstein, significantly increased the xylose content. However, no significant relationship was observed between the liquid hydrolysate xylose content and the glucose content obtained from enzymatic saccharification of the acid-insoluble residue. These results highlight the potential of wheat breeding to improve biomass-related traits in wheat straw.

Molecular and Cytogenetic Characterization of the 5DS-5BS Chromosome Translocation Conditioning Soft Kernel Texture in Durum Wheat.

The durum wheat ( ssp. (Desf.) Husn.) cultivar Soft Svevo with a soft kernel texture was developed through a -mediated homoeologous 5DS-5BS chromosomal translocation. The soft kernel trait ( locus) derived from chromosome 5D of the common wheat ( L.) cultivar Chinese Spring. Soft Svevo was used as the donor parent to create near-isogenic soft durum germplasm. The size of the translocation, its estimated breakpoint, and the amount of chromosome 5BS translocated, if any, remain unknown. Four near-isogenic pairs of hard and soft kernel durum genotypes, in addition to Soft Svevo and the Chinese Spring deletion line 5DS-2, which lacks a distal 22% terminal segment of chromosome 5DS, were genotyped using Illumina's 90k wheat single nucleotide polymorphism array. Single nucleotide polymorphism results were processed in GenomeStudio and 164 polymorphic markers were identified between the near-isogenic lines (NILs). Subsequent BLASTn results for two subsets of markers corresponding to the distal ends of chromosomes 5DS and 5BS indicated that the translocation event was nearly reciprocal, as a ∼24.36-Mbp segment of chromosome 5DS was gained, whereas a ∼20.01-Mbp segment of chromosome 5BS was lost. Genomic in situ hybridization images of the soft durum NILs agreed with these estimates and confirmed the absence of additional terminal or interstitial translocations. Soft durum represents the potential of a new wheat market class and these findings will assist durum wheat breeders in the development of new soft durum germplasm.

Aegilops tauschii Accessions with Geographically Diverse Origin Show Differences in Chromosome Organization and Polymorphism of Molecular Markers Linked to Leaf Rust and Powdery Mildew Resistance Genes.

Aegilops tauschii (2n = 2x = 14) is a diploid wild species which is reported as a donor of the D-genome of cultivated bread wheat. The main goal of this study was to examine the differences and similarities in chromosomes organization among accessions of Ae. tauschii with geographically diversed origin, which is believed as a potential source of genes, especially determining resistance to fungal diseases (i.e., leaf rust and powdery mildew) for breeding of cereals. We established and compared the fluorescence in situ hybridization patterns of 21 accessions of Ae. tauschii using various repetitive sequences mainly from the BAC library of wheat cultivar Chinese Spring. Results obtained for Ae. tauschii chromosomes revealed many similarities between analyzed accessions, however, some hybridization patterns were specific for accessions, which become from cognate regions of the World. The most noticeable differences were observed for accessions from China which were characterized by presence of distinct signals of pTa-535 in the interstitial region of chromosome 3D, less intensity of pTa-86 signals in chromosome 2D, as well as lack of additional signals of pTa-86 in chromosomes 1D, 5D, or 6D. Ae. tauschii of Chinese origin appeared homogeneous and separate from landraces that originated in western Asia. Ae. tauschii chromosomes showed similar hybridization patterns to wheat D-genome chromosomes, but some differences were also observed among both species. What is more, we identified reciprocal translocation between short arm of chromosome 1D and long arm of chromosome 7D in accession with Iranian origin. High polymorphism between analyzed accessions and extensive allelic variation were revealed using molecular markers associated with resistance genes. Majority of the markers localized in chromosomes 1D and 2D showed the diversity of banding patterns between accessions. Obtained results imply, that there is a moderate or high level of polymorphism in the genome of Ae. tauschii determined by a geographical origin, which we proved by cytogenetic and molecular markers analysis. Therefore, selected accessions might constitute an accessible source of variation for improvement of Triticeae species like wheat and triticale.

Pattern of Protein Expression in Developing Wheat Grains Identified through Proteomic Analysis.

Grain development is one of the biological processes, which contributes to the final grain yield. To understand the molecular changes taking place during the early grain development, we profiled proteomes of two common wheat cultivars P271 and Chinese Spring (CS) with large and small grains, respectively at three grain developmental stages (4, 8, and 12 days post anthesis). An iTRAQ (isobaric tags for relative and absolute quantitation) based proteomics approach was used for this purpose. More than 3,600 proteins were reported to accumulate during early grain development in both wheat cultivars. Of these 3,600 proteins, 130 expressed differentially between two wheat cultivars, and 306 exhibited developmental stage-specific accumulation in either or both genotypes. Detailed bioinformatic analyses of differentially expressed proteins (DEPs) from the large- and small-grain wheat cultivars underscored the developmental differences observed between them and shed light on the molecular and cellular processes contributing to these differences. In silico localization of either or both sets of DEPs to wheat chromosomes exhibited a biased genomic distribution with chromosome 4D contributing largely to it. These results corresponded well with the earlier studies, performed in common wheat, where chromosome 4D was reported to harbor QTLs for yield contributing traits specifically grain length. Collectively, our results provide insight into the molecular processes taking place during early grain development, a knowledge, which may prove useful in improving wheat grain yield in the future.

Zn and Fe Concentration Variations of Grain and Flag Leaf and the Relationship with NAM-G1 Gene in Triticum timopheevii (Zhuk.) Zhuk. ssp. timopheevii

Grains of 12 accessions of Triticum timopheevii (Zhuk.) Zhuk. ssp. timopheevii (AAGG, 2n = 4x = 28) and one bread wheat cultivar Chinese Spring (CS) and one durum wheat cultivar Langdon (LDN) grown across two years were analyzed for grain iron (Fe) and zinc (Zn) concentrations. All the 12 tested T. timopheevii ssp. timopheevii genotypes showed significantly higher concentration of grain Fe and Zn than CS and LDN. Aboundant genetic variability of both the Fe and Zn concentrations was observed among the T. timopheevii ssp. timopheevii accessions, averagely varied from 47.06 to 90.26 mg kg−1 and from 30.05 to 65.91 mg kg−1, respectively. Their grain Fe and Zn concentrations between years exhibited a significantly positive correlation with the correlation coefficients r = 0.895 and r = 0.891, respectively, indicating the highly genetic stability. Flag leaf possessed twice or three times higher concentrations for both Fe and Zn than grain, and a significantly high positive correlation appeared between the two organs with r = 0.648 for Fe and r = 0.957 for Zn concentrations, respectively, suggesting flag leaves might be indirectly used for evaluating grain Zn and Fe contents. Significant correlations occurred between grain Fe and Zn concentrations, and between grain Zn concentration and the two agronomic traits of plant height and number of spikelets per spike. Both the concentrations were not related to seed size or weight as well as NAM-G1 gene, implying the higher grain Fe and Zn concentrations of T. timopheevii ssp. timopheevii species are not ascribed to concentration effects of seed and the genetic control of NAM-G1 gene. There might be some other biological factors impacting the grain’s Zn and Fe concentrations. These results indicated T. timopheevii ssp. timopheevii species might be a promising genetic resource with high Fe and Zn concentrations for the biofortification of current wheat cultivars.

A novel compensating wheat-Thinopyrum elongatum Robertsonian translocation line with a positive effect on flour quality.

Wheat flours are used to produce bread, pasta, breakfast cereals, and biscuits; the various properties of these end-products are attributed to the gluten content, produced as seed storage proteins in the wheat endosperm. Thus, genes encoding gluten protein are major targets of wheat breeders aiming to improve the various properties of wheat flour. Here, we describe a novel compensating wheat–Thinopyrum elongatum Robertsonian translocation (T1AS.1EL) line involving the short arm of wheat chromosome 1A (1AS) and the long arm of Th. elongatum chromosome 1E (1EL); we developed this line through centric breakage-fusion. Compared to the common wheat cultivars Chinese Spring and Norin 61, we detected two additional 1EL-derived high-molecular-weight glutenin subunits (HMW-GSs) in the T1AS.1EL plants. Based on the results of an SDS-sedimentation volume to estimate the gluten strength of T1AS.1EL-derived flour, we predict that T1AS.1EL-derived flour is better suited to bread-making than Chinese Spring- and Norin 61-derived flour and that this is because of its greater gluten diversity. Also, we were able to assign 33 of 121 wheat PCR-based Landmark Unique Gene markers to chromosome 1E of Th. elongatum. These markers can now be used for further chromosome engineering of the Th. elongatum segment of T1AS.1EL.

Characterization of proteins involved in early stage of wheat grain development by iTRAQ

Wheat (Triticum aestivum L.) is one of the most important crops in the world due to its value as a major food source. To explore the protein synthesis, accumulation and regulation during grain development are more complex because of its large genome. In this study, grains from wheat cultivar Chinese Spring were harvested at three development stages (DAA4, DAA8 and DAA12) and were used to display variable expressed proteins during these stages. iTRAQ-based quantitative proteomic and LC–MS/MS methods were used in this study. A total of 1762 proteins showed that at least 1.5-fold differences in abundance were found during these stages; most of the differential expressed proteins (DEPs) were involved in the metabolism process, which includes carbohydrate metabolism, cell division, cytoskeleton, lipid metabolism, nitrogen metabolism, protein synthesis, signal transduction, translation and transport. The results provide insight into biochemical events taking place during the early wheat grain development and highlight the value of proteomics in characterizing complex biochemical processes. Our results also showed that the photosynthesis played an important role for implying energy during the early grain development. SignificanceWheat (T. aestivum L., 2n=6X=42, AABBDD) is one of the most important crops in the world. The early phase of seed grain development mainly involved active cell enlargement, leading to a rapid increase in seed size available for further accumulation of starch of storage proteins. We applied the iTRAQ proteomic analysis to decipher the mechanism of wheat grain development during the early stage. Numerous differentially expressed proteins were identified; most of which were related to carbohydrate metabolism, cell division, cytoskeleton, lipid metabolism, nitrogen metabolism, protein synthesis, signal transduction, translation and transport. Results also showed that the photosynthesis played an important role for implying energy during the early grain development. Taken together, our results provide comprehensive proteome insights into the early wheat grain development.

Development and Validation of Thinopyrum elongatum–Expressed Molecular Markers Specific for the Long Arm of Chromosome 7E

ABSTRACTThe ditelocentric addition line CS‐7EL of the spring wheat (Triticum aestivum L.) cultivar Chinese Spring (CS) contains the long arm of the chromosome 7E from Thinopyrum elongatum (CS‐7EL), which confers high resistance to Fusarium head blight. It is of great interest to breeders to integrate the resistance locus (loci) from Th. elongatum into commercial wheat varieties. The objectives of this study were to identify candidate genes expressed from the 7EL chromosome of CS‐7EL, to develop 7EL‐specific molecular markers, and to validate their usefulness for characterizing recombination between one of the group 7 chromosomes of wheat and Th. elongatum. High‐throughput sequencing of Fusarium graminearum–infected and control CS and CS‐7EL cDNA libraries was performed using RNA‐Seq. A stepwise bioinformatics strategy was applied to assemble the sequences obtained from RNA‐Seq and to create a conservative list of candidate genes expressed from the foreign chromosome 7EL. Polymerase chain reaction primer pairs were designed and tested for 135 candidate genes. A total of 48 expressed molecular markers specific for the chromosome 7EL were successfully developed. Screening of progenies from two BC1F2 families from the cross CS‐7E(7D) × 2*CSph1b showed that these markers are useful for characterizing recombination events between the chromosomes 7D from wheat and 7E from Th. elongatum.

Identification of new heading date determinants in wheat 5B chromosome.

BackgroundVariability of heading date may assist in wheat adaptation to local environments. Thereafter, discovery of new heading date determinants is important for cereal improvement. In this study we used common wheat cultivar Chinese Spring (CS) and the substitution line of CS with 5B chromosome from T. dicoccoides (CS-5Bdic), different in their heading date by two weeks, to detect determinants of heading date on 5B chromosome.ResultsThe possible influence of the VRN-B1 gene, the most powerful regulator of flowering, located on 5B chromosome, to differences in heading time between CS and CS-5Bdic was studied. The sequencing of this gene from CS-5Bdic showed that an insertion of a nucleotide triplet produced an additional amino acid in the corresponding protein. No changes in the transcription levels of each homoeologous VRN-1 loci were found in CS-5Bdic by comparison with CS. To ascertain the loci determining heading date difference, a set of 116 recombinant inbred 5В chromosomal lines as a result of hybridization of CS with CS-5Bdic were developed and their heading dates were estimated. Using the Illumina Infinium 15 k Wheat platform, 379 5B-specific polymorphic markers were detected and a genetic map with 82 skeletal markers was constructed. Phenotype (heading date) – genotype association analysis revealed seventy eight markers in pericentromeric region of 5B chromosome significantly associated with heading date variation. Based on this estimation and synteny with model crop genomes we identified the three best candidate genes: WRKY, ERF/AP2 and FHY3/FAR1.ConclusionsWe supposed that the difference in activity of WRKY, ERF/AP2 and/or FHY3/FAR1 transcription factors between CS and CS-5Bdic to be a probable reason for the observed difference in heading dates. Data obtained in this study provide a good basis for the subsequent investigation of heading time pathways in wheat.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0688-x) contains supplementary material, which is available to authorized users.