Search

Chinese wheat landrace Anyuehong (AYH) has displayed high levels of stable adult plant resistance (APR) to stripe rust for >15 years. To identify quantitative trait loci (QTLs) for stripe rust resistance in AYH, a set of 110 recombinant inbred lines (RILs) was developed from a cross between AYH and susceptible cultivar Taichung 29. The parents and RILs were evaluated for final disease severity (FDS) in six field tests with a mixture of predominant Puccinia striiformis f. sp. tritici races at the adult plant stage and genotyped via the wheat 55K single-nucleotide polymorphism (SNP) array to construct a genetic map with 1,143 SNP markers. Three QTLs, designated as QYr.AYH-1AS, QYr.AYH-5BL, and QYr.AYH-7DS, were mapped on chromosome 1AS, 5BL, and 7DS, respectively. RILs combining three QTLs showed significantly lower FDS compared with the lines in other combinations. Of them, QYr.AYH-5BL and QYr.AYH-7DS were stably detected in all environments, explaining 13.6 to 21.4% and 17.6 to 33.6% of phenotypic variation, respectively. Compared with previous studies, QYr.AYH-5BL may be a new QTL, whereas QYr.AYH-7DS may be Yr18. Haplotype analysis revealed that QYr.AYH-5BL is probably present in 6.2% of the 323 surveyed Chinese wheat landraces. The kompetitive allele specific PCR (KASP) markers for QYr.AYH-5BL were developed by the linked SNP markers to successfully confirm the effects of the QTL in a validation population derived from a residual heterozygous line and were further assessed in 38 Chinese wheat landraces and 92 cultivars. Our results indicated that QYr.AYH-5BL with linked KASP markers has potential value for marker-assisted selection to improve stripe rust resistance in breeding programs.

Summary Pre‐harvest sprouting (PHS), the germination of grain before harvest, is a serious problem resulting in wheat yield and quality losses.Here, we mapped the PHS resistance gene PHS‐3D from synthetic hexaploid wheat to a 2.4 Mb presence–absence variation (PAV) region and found that its resistance effect was attributed to the pleiotropic Myb10‐D by integrated omics and functional analyses.Three haplotypes were detected in this PAV region among 262 worldwide wheat lines and 16 Aegilops tauschii, and the germination percentages of wheat lines containing Myb10‐D was approximately 40% lower than that of the other lines. Transcriptome and metabolome profiling indicated that Myb10‐D affected the transcription of genes in both the flavonoid and abscisic acid (ABA) biosynthesis pathways, which resulted in increases in flavonoids and ABA in transgenic wheat lines. Myb10‐D activates 9‐cis‐epoxycarotenoid dioxygenase (NCED) by biding the secondary wall MYB‐responsive element (SMRE) to promote ABA biosynthesis in early wheat seed development stages.We revealed that the newly discovered function of Myb10‐D confers PHS resistance by enhancing ABA biosynthesis to delay germination in wheat. The PAV harboring Myb10‐D associated with grain color and PHS will be useful for understanding and selecting white grained PHS resistant wheat cultivars.

A novel, stably expressed, and plant height-independent QTL for spike extension length on 5AS was identified and validated in different populations using a newly developed and tightly linked KASP marker. As an important component of plant height (PH), spike extension length (SEL) plays a significant role in formation of an ideotype in wheat. Despite the fact that numerous loci for SEL in wheat have been reported, our knowledge on PH-independent loci remains to be limited. In this study, two recombinant inbred line (RIL) populations genotyped using the Wheat55K SNP were used to detect quantitative trait loci (QTL) controlling SEL across six environments. A total of 30 QTL for SEL were detected in these two RIL populations, and four of them, i.e., QSEL.sicau-2CN-4D, QSEL.sicau-2SY-4B.2, QSEL.sicau-2SY-4D.1, and QSEL.sicau-2CN-5A, were stably expressed. Genetic and conditional QTL analysis showed that the first three were significantly associated with PH, while the last one, QSEL.sicau-2CN-5A, is independent of PH. Comparison of genetic and physical maps suggested that only QSEL.sicau-2CN-5A located on chromosome arm 5AS is likely a novel QTL. A Kompetitive Allele-Specific PCR (KASP) marker, KASP-AX-110413733, tightly linked to this novel QTL was developed to successfully confirm its effect in three different genetic populations. Further, in the interval where QSEL.sicau-2CN-5A was located on 'Chinese Spring' wheat reference genome, three promising genes mainly expressed in wheat stem were predicated and they all encode the cytochrome P450 that was demonstrated to be closely associated with SEL elongation in rice. In addition, significant correlations between SEL and PH, spikelet number per spike, and thousand-grain weight were also detected. Altogether, our results broaden our understanding on genetic basis of SEL and will be useful for marker-based selection of lines with different SELs and fine mapping the novel and PH-independent QTL QSEL.sicau-2CN-5A.

In our previous work, a novel high-molecular-weight glutenin subunit (HMW-GS) with an extremely large molecular weight from Aegilops sharonensis was identified that may contribute to excellent wheat (Triticum aestivum) processing quality and increased dough strength, and we further generated HMW-GS homozygous lines by crossing. In this study, we crossed the HMW-GS homozygous line 66-17-52 with ‘Chinese Spring’ Ph1 mutant CS ph1b to induce chromosome recombination between wheat and Ae. sharonensis. SDS-PAGE was used to identify 19 derived F2 lines with the HMW-GSs of Ae sharonensis. The results of non-denaturing fluorescence in situ hybridization (ND-FISH) indicated that lines 6-1 and 6-7 possessed a substitution of both 5D chromosomes by a pair of 1Ssh chromosomes. Further verification by newly developed 1Ssh-specific chromosome markers showed that these two lines amplified the expected fragment. Thus, it was concluded that lines 6-1 and 6-7 are 1Ssh(5D) chromosome substitution lines. The 1Ssh(5D) chromosome substitution lines, possessing alien subunits with satisfactory quality-associated structural features of large repetitive domains and increased number of subunits, may have great potential in strengthening the viscosity and elasticity of dough made from wheat flour. Therefore, these substitution lines can be used for wheat quality improvement and further production of 1Ssh translocation lines.

Stripe rust (caused by Puccinia striiformis f. sp. tritici) is one of the most severe diseases affecting wheat production. The disease is best controlled by developing and growing resistant cultivars. Chinese wheat (Triticum aestivum) landraces have excellent resistance to stripe rust. The objectives of this study were to identify wheat landraces with stable resistance and map quantitative trait loci (QTL) for resistance to stripe rust from 271 Chinese wheat landraces using a genome-wide association study (GWAS) approach. The landraces were phenotyped for stripe rust responses at the seedling stage with two predominant Chinese races of P. striiformis f. sp. tritici in a greenhouse and the adult-plant stage in four field environments and genotyped using the 660K wheat single-nucleotide polymorphism (SNP) array. Thirteen landraces with stable resistance were identified, and 17 QTL, including eight associated to all-stage resistance and nine to adult-plant resistance, were mapped on chromosomes 1A, 1B, 2A, 2D, 3A, 3B, 5A, 5B, 6D, and 7A. These QTL explained 6.06–16.46% of the phenotypic variation. Five of the QTL, QYrCL.sicau-3AL, QYrCL.sicau-3B.4, QYrCL.sicau-3B.5, QYrCL.sicau-5AL.1 and QYrCL.sicau-7AL, were likely new. Five Kompetitive allele specific PCR (KASP) markers for four of the QTL were converted from the significant SNP markers. The identified wheat landraces with stable resistance to stripe rust, significant QTL, and KASP markers should be useful for breeding wheat cultivars with durable resistance to stripe rust.

Wheat is a major staple food crop worldwide because of the unique properties of wheat flour. High molecular weight glutenin subunits (HMW-GSs), which are among the most critical determinants of wheat flour quality, are responsible for the formation of glutenin polymeric structures via interchain disulfide bonds. We herein describe the identification of a new HMW-GS Dy10 allele (Dy10-m619SN). The amino acid substitution (serine-to-asparagine) encoded in this allele resulted in a partial post-translational cleavage that produced two new peptides. These new peptides disrupted the interactions among gluten proteins because of the associated changes to the number of available cysteine residues for interchain disulfide bonds. Consequently, Dy10-m619SN expression decreased the size of glutenin polymers and weakened glutens, which resulted in wheat dough with improved cookie-making quality, without changes to the glutenin-to-gliadin ratio. In this study, we clarified the post-translational processing of HMW-GSs and revealed a new genetic resource useful for wheat breeding.

Additional file 2. Pearson’s correlation coefficients for infection type (IT), final disease severity (FDS) and area under the disease progress curve (AUDPC) against stripe rust evaluated among five environments during 2016 to 2018. Different environments were all correlated, Significant at P

Additional file 6. The avirulence(A) /virulence(V) formula of the Pst races used in this study.

Increasing wheat yield is essential in solving the global food shortage. The spikelet number per spike (SNS) is a primary factor that determines the kernel number per spike which has a great effect on wheat yield. Modern wheat breeding methods reduce the genetic diversity among cultivars, leading to a detrimental effect on future wheat production. It is, therefore, necessary to explore new genetic resources for SNS to increase wheat yield. A tetraploid landrace Ailanmai wild emmer wheat recombinant inbred line (RIL) population was used to construct a high-density genetic map using the wheat 55K single nucleotide polymorphism (SNP) array. The linkage map revealed that 94.83% (6204) of the mapped markers had consistent genetic and physical chromosomal locations. Fourteen quantitative trait loci (QTL) for SNS, explaining 4.23-27.26% of the phenotypic variation, were identified. QSns.sau-AM-2B.3 and QSns.sau-AM-3B.2 were detected as the major and novel QTL with a large cumulative effect that increased SNS by 17.47%. Some development-related genes within the physical intervals of QSns.sau-AM-2B.3 and QSns.sau-AM-3B.2 were predicted to participate in spikelet growth, thereby affecting SNS. Genetic associations between SNS and other agronomic traits such as a significant and positive correlation between SNS and thousand kernel weight (TKW) were detected and analyzed. QTL analysis was also performed for other spike-related traits, including spike length (SL), kernel number per spike (KNS), kernel number per spikelet (KNL), and spike density (SD). QSns.sau-AM-2B.3 was co-located with QSl.sau-AM-2B in the interval between AX-111626925 and AX-111588559. Similarly, QKns.sau-AM-5B.1 was co-related with QKnl.sau-AM-5B.2 in the region between AX-108760512 and AX-110180251. This study demonstrates the feasibility of wheat 55K SNP array in genetic mapping of tetraploid wheat population and shows the potential application of wheat related species in wheat improvement programs.

Stripe rust (yellow rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Chinese wheat landrace Guangtoumai (GTM) exhibited a high level of resistance against predominant P. striiformis f. sp. tritici races in China at the adult plant stage. The objective of this research was to identify and map the major locus/loci for stripe rust resistance in GTM. A set of 212 recombinant inbred lines (RILs) was developed from a cross between GTM and Avocet S. The parents and RILs were evaluated in three field tests (2018, 2019, and 2020 at Chongzhou, Sichuan) with the currently predominant P. striiformis f. sp. tritici races for final disease severity and genotyped with the Wheat 55K single nucleotide polymorphism (SNP) array to construct a genetic map with 1,031 SNP markers. A major locus, named QYr.GTM-5DL, was detected on chromosome 5DL in GTM. The locus was mapped in a 2.75-cM interval flanked by SNP markers AX-109855976 and AX-109453419, explaining up to 44.4% of the total phenotypic variation. Since no known Yr genes have been reported on chromosome 5DL, QYr.GTM-5DL is very likely a novel adult plant resistance locus. Haplotype analysis revealed that the resistance allele displayed enhanced levels of stripe rust resistance and is likely present in 5.3% of the 247 surveyed Chinese wheat landraces. The derived cleaved amplified polymorphic sequence (dCAPS) marker dCAPS-5722, converted from a SNP marker tightly linked to QYr.GTM-5DL with 0.3 cM, was validated on a subset of RILs and 48 commercial wheat cultivars developed in Sichuan. The results indicated that QYr.GTM-5DL with its linked dCAPS marker could be used in marker-assisted selection to improve stripe rust resistance in breeding programs, and this quantitative trait locus will provide new and possibly durable resistance to stripe rust.

Background Stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst), is a serious foliar disease of wheat. Identification of novel stripe rust resistance genes and cultivation of resistant cultivars are considered to be the most effective approaches to control this disease. In this study, we evaluated the infection type (IT), disease severity (DS) and area under the disease progress curve (AUDPC) of 143 Chinese wheat landrace accessions for stripe rust resistance. Assessments were undertaken in five environments at the adult-plant stage with Pst mixture races under field conditions. In addition, IT was assessed at the seedling stage with two prevalent Pst races (CYR32 and CYR34) under a controlled greenhouse environment. Results Seventeen accessions showed stable high-level resistance to stripe rust across all environments in the field tests. Four accessions showed resistance to the Pst races CYR32 and CYR34 at the seedling stage. Combining phenotypic data from the field and greenhouse trials with 6404 markers that covered the entire genome, we detected 17 quantitative trait loci (QTL) on 11 chromosomes for IT associated with seedling resistance and 15 QTL on seven chromosomes for IT, final disease severity (FDS) or AUDPC associated with adult-plant resistance. Four stable QTL detected on four chromosomes, which explained 9.99–23.30% of the phenotypic variation, were simultaneously associated with seedling and adult-plant resistance. Integrating a linkage map of stripe rust resistance in wheat, 27 QTL overlapped with previously reported genes or QTL, whereas four and one QTL conferring seedling and adult-plant resistance, respectively, were mapped distantly from previously reported stripe rust resistance genes or QTL and thus may be novel resistance loci. Conclusions Our results provided an integrated overview of stripe rust resistance resources in a wheat landrace diversity panel from the southern autumn-sown spring wheat zone of China. The identified resistant accessions and resistance loci will be useful in the ongoing effort to develop new wheat cultivars with strong resistance to stripe rust.

Anthocyanins and selenium have vital biological functions for human and plants, they were investigated thoroughly and separately in plants. Previous studies indicated pigmented fruits and vegetables had higher selenium concentration, but whether there is a relationship between anthocyanins and selenium is unclear. In this study, a combined phenotypic and genotypic methodological approach was undertaken to explore the potential relationship between anthocyanins and selenium accumulation by using phenotypic investigation and RNA-seq analysis. The results showed that pigmented cultivars enrichment in Se is a general phenomenon observed for these tested species, this due to pigmented cultivars have higher Se efficiency absorption. Se flow direction mainly improve concentration of S-rich proteins of LMW-GS. This may be a result of the MYB and bHLH co-regulate anthocyanins biosynthesis and Se metabolism at the transcriptional level. This thesis addresses a neglected aspect of the relevant relationship between anthocyanins and selenium.

Eight major and stably expressed QTL for flag leaf morphology across eleven environments were identified and validated using newly developed KASP markers in seven biparental populations with different genetic backgrounds. Flag leaf morphology is a determinant trait influencing plant architecture and yield potential in wheat (Triticum aestivum L.). A recombinant inbred line (RIL) population with a 55 K SNP-based constructed genetic map was used to map quantitative trait loci (QTL) for flag leaf length (FLL), width (FLW), area (FLA), angle (FLANG), opening angle (FLOA), and bend angle (FLBA) in eleven environments. Eight major QTL were detected in 11 environments with 5.73–54.38% of explained phenotypic variation. These QTL were successfully verified using the newly developed Kompetitive Allele Specific PCR (KASP) markers in six biparental populations with different genetic backgrounds. Among these 8 major QTL, two co-located intervals were identified. Significant interactions for both FLL- and FLW-related QTL were detected. Comparison analysis showed that QFll.sau-SY-2B and QFla.sau-SY-2B are likely new loci. Significant relationships between flag leaf- and yield-related traits were observed and discussed. Several genes associated with leaf development including the ortholog of maize ZmRAVL1, a B3-domain transcription factor involved in regulation of leaf angle, were predicted in physical intervals harboring these major QTL on reference genomes of bread wheat ‘Chinese spring’, T. turgidum, and Aegilops tauschii. Taken together, these results broaden our understanding on genetic basis of flag leaf morphology and provide clues for fine mapping and marker-assisted breeding wheat with optimized plant architecture for promising loci.

Chinese endemic wheat, comprising Tibetan semi-wild wheat (Triticum aestivum ssp. tibetanum), Yunnan hulled wheat (T. aestivum ssp. yunnanense), and Xinjiang rice wheat (T. petropavlovskyi), are genetically and morphologically unique. To examine the adult plant resistance to stripe rust among Chinese endemic wheat germplasms, a panel of 213 accessions was inoculated with mixed virulent races of wheat stripe rust (Puccinia striiformis f. sp. tritici) in four different field environments. Four traits associated with stripe rust resistance, infection type, final disease severity, disease index, and area under the disease progress curve, were used to evaluate the accessions. The phenotypic datasets were used for 55K single-nucleotide polymorphism (SNP) array-based genome-wide association studies to identify effective resistance loci. Eighty-nine accessions with stable resistance were identified in at least three of the four environments by phenotypic evaluation. Eleven markers located on chromosomes 1A, 2B, 5A, 5D, 7B, and 7D by the genome-wide association studies analysis showed significant associations with at least two resistance-associated traits in two of the environments. These loci, corresponding to seven genomic regions based on linkage disequilibrium decay distance, explained 9.3 to 26.0% of the total phenotypic variation. Five quantitative trait loci (QTLs) on chromosomes 1A, 2B, 7B, and 7D overlapped or were in close proximity to previously reported QTLs based on the consensus and physical maps using the reference sequence of bread wheat (IWGSC RefSeq v1.0). The other two QTLs were potential novel QTLs given their physical positions. Haplotype variants of QTL QYr.sicau-2BS showed subspecies-specific inheritance of the stripe rust resistance locus. Resistant loci among Chinese endemic wheat germplasms could be introduced into common wheat cultivars, and the high-confidence SNP markers will aid in marker-assisted selection in breeding for stripe rust disease resistance.

In this study, we successfully expressed the active 1Ay subunit of Triticum urartu in barley by Agrobacterium-mediated transformation with a transformation efficiency of 19.9%. The results of SDS-PAGE revealed that the expressed proteins of 1Ay subunit were present at some grains of each of 46 original T0 plants, showing identical mobility to those of positive standards of T. urartu grain protein and bacteria expressional proteins. In the T2 generation, three homozygous lines, 2-28, 3-11, and 5-6, were identified. The results of scanning electron microscopy showed an increased amount of protein bodies in these transgenic lines. The main effects in the expression of the 1Ay subunits was a considerable increase in the glutenin content, but a decrease in the contents of gliadins while there were no effects in the contents of albumin, globulin and the total protein. We found that the gluten could not be washed out from the flour obtained from transgenic barley lines when using a Gluten index analyzer and a Farinograph indicating that the transgenic barley lines could not form dough. The lack of x-type HMW-GS and the reduction in number of subunit were inferred as the possible reasons for the inability to form gluten polymer.

Background:Stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst), is a serious foliar disease of wheat. Identification of novel stripe-rust resistance genes and cultivation of resistant varieties are considered to be the most effective approaches to control this disease. In this study, we evaluated the infection type (IT), disease severity (DS) and area under the disease progress curve (AUDPC) of 143 Chinese wheat landrace accessions for stripe-rust resistance. Assessments were undertaken in five environments at the adult-plant stage with Pst mixture races under field conditions, in addition, IT was assessed at the seedling stage with two prevalent Pst races (CYR32 and CYR34) under controlled greenhouse conditions. Results:Seventeen accessions showed stable high-level resistance to stripe rust across all environments under field tests,while four accessions showed resistance to the Pst races CYR32 and CYR34 at the all-stage.Combining phenotypic data from the field and greenhouse trials with 6404 markers covering the whole genome, we detected 17quantitative trait locus(QTLs) on 11 chromosomes for IT associated with seedling resistance and 15 QTLs on seven chromosomes for IT, final disease severityor AUDPC associated with adult-plant resistance. Four stable QTLs detected on four chromosomes, which explained 9.99%–23.30% of the phenotypic variation, were simultaneously associated with seedling and adult-plant resistance. Integrating a linkage map of stripe-rust resistance in wheat, 29 QTLs overlapped with previously reported genes or QTLs, while two and oneQTLs conferring seedling and adult-plant resistancerespectively were mapped distantly from previously reported stripe rust resistance genes or QTLs and may be novel resistantloci. Conclusions:Our results provided an integrated view of stripe rust resistance resources in a Chinese wheat landrace diversity panel from the southern autumn-sown spring wheat zone.The identified resistant accessions and resistance loci will be useful in the ongoing effort to develop new wheat cultivars with strong resistance to stripe rust.

In our previous study, we have isolated novel HMW-GSs with unusually large molecular weight from Aegilops sharonensis; and to introduce this novel HMW-GSs variant into common wheat, we produced a number of amphiploids between various tetraploid wheat, hexaploid wheat and Ae. sharonensis by distant hybridization. In this study, we reported that the characterization of 24 HMW-GSs homozygous lines from wheat-Ae. sharonensis populations derived by continuous backcross or self-cross to common wheat cultivar LM3, which possessed homozygous HMW-GSs compositions including alien HMW-GS of Ae. sharonensis plus B and/or D genomes encoded HMW-GSs of LM3. According to the profiling of HMW-GS, these 24 homozygous lines can be summarized as four types. The results of Non-denaturing fluorescence in situ hybridization (ND-FISH) indicated that the chromosome numbers of eight of 24 lines were 40, 42, 44 or 45, and the number of alien Ae. sharonensis Ssh were between two and eight. Three 1S(1A) substitution lines, two 1S(1B) substitution lines, three 1S(1D) substitution lines. For the four lines with 44 and 45 chromosomes, they exhibit novel cytogenetic characteristics in which the substitution of 1S(1B) or 1S(1D) and addition of Ae. sharonensis 3S or 4S simultaneously appeared. The 24 homozygous progenies were similar to common wheat LM3 on agronomic traits. Specially, four lines of them showed almost identical agronomic traits to recurrent parent common wheat LM3 than the rest lines. These homozygous progenies could be used not only to evaluate function on quality of single HMW-GS but also to create translocation lines.

As an essential mineral element, selenium (Se) plays a critical role in human health. Given the low concentrations ( stem > grain. This is the first study to document a Se-rich synthetic wheat line, and root structure and Se grain concentration was strongly affected by QTL located on 3D.

Stripe rust, which is caused by the biotrophic fungus Puccinia striiformis f. sp. tritici (Pst), is one of the most challenging fungal diseases affecting wheat (Triticum aestivum L.), because specific virulent races often overcome host resistance genes. There is a continuous demand for new host resistance genes that are effective against such races. Chinese wheat landrace, which is the primary gene pool of the common wheat, is a valuable source of novel stripe rust resistance genes for wheat breeding. Chinese wheat landrace ‘Hejiangyizai’ (HY) is highly resistant to a wide spectrum of Chinese Pst races in both seedling and adult-plant stages. Genetic analysis has indicated that a single dominant gene in HY confers resistance to stripe rust, which has been temporarily designated as YrHY. Segregation of the F2 population and its F2:3 families derived from the cross between HY and Taichung 29 was used for bulked segregant analysis using simple sequence repeats (SSRs), resistance gene analog polymorphisms (RGAPs), sequence-related amplified polymorphisms (SRAPs), and target region amplified polymorphisms (TRAPs). Linkage analysis indicated that six SSR, eight RGAP, two TRAP, and six SRAP polymorphic markers were linked to the stripe rust resistance gene YrHY. The two closest flanking SSR markers were Xbarc92 and Xcfd66, with genetic distances of 4.3 and 8.5 cM, respectively. YrHY was physically mapped to chromosome 7DS using a set of Chinese spring nullisomic-tetrasomic lines. Comprehensive analyses of the seedling and adult reaction patterns of HY tested whit Chinese PST race CYR32, allelic test, and pedigree relationship indicated that YrHY differed from Yr18, Yr33, YrY201, YrY212, and YrWV on chromosome 7D. These results demonstrated that the YrHY may be a novel stripe rust resistance gene.

Background Kernel length (KL), kernel width (KW) and thousand-kernel weight (TKW) are key agronomic traits in wheat breeding. Chuannong16 (‘CN16’) is a commercial cultivar with significantly longer kernels than the line ‘20828’. To identify and characterize potential alleles from CN16 controlling KL, the previously developed recombinant inbred line (RIL) population derived from the cross ‘20828’ × ‘CN16’ and the genetic map constructed by the Wheat55K SNP array and SSR markers were used to perform quantitative trait locus/loci (QTL) analyses for kernel traits. Results A total of 11 putative QTL associated with kernel traits were identified and they were located on chromosomes 1A (2 QTL), 2B (2 QTL), 2D (3 QTL), 3D, 4A, 6A, and 7A, respectively. Among them, three major QTL, QKL.sicau-2D, QKW.sicau-2D and QTKW.sicau-2D, controlling KL, KW and TKW, respectively, were detected in three different environments. Respectively, they explained 10.88–18.85%, 17.21–21.49% and 10.01–23.20% of the phenotypic variance. Further, they were genetically mapped in the same interval on chromosome 2DS. A previously developed kompetitive allele-specific PCR (KASP) marker KASP-AX-94721936 was integrated in the genetic map and QTL re-mapping finally located the three major QTL in a 1- cM region flanked by AX-111096297 and KASP-AX-94721936. Another two co-located QTL intervals for KL and TKW were also identified. A few predicted genes involved in regulation of kernel growth and development were identified in the intervals of these identified QTL. Significant relationships between kernel traits and spikelet number per spike and anthesis date were detected and discussed. Conclusions Three major and stably expressed QTL associated with KL, KW, and TKW were identified. A KASP marker tightly linked to these three major QTL was integrated. These findings provide information for subsequent fine mapping and cloning the three co-localized major QTL for kernel traits.

In this study, we successfully knock-out the d-hordein component of barley storage protein using RNA-guided Cas9. Mutation frequencies of 25% and 14% at two different target sites were obtained. Homozygous mutant plants that were T-DNA free were identified in the T1 generation. Barley grains without d-hordein proteins from T2 seeds showed a significantly reduced grain size compared to the parent plant and control non-edited line. The protein matrix surrounding the starch granules was increased, whereas the starch granules themselves were decreased in size in the mutant plants compared to controls. The main effect of a lack of d-hordein was a considerable decrease in the prolamines and an increase in the glutenins. The changes of other grain composition included the increased starch content, amylose content, and β-glucan content. The roles of d-hordein mutation on barley grain size and grain composition remain to be studied.

Terpenoids play important roles in plant defense. Although some terpene synthases have been characterized, terpenoids and their biosynthesis in wheat (Triticum aestivum L.) still remain largely unknown. Here, we describe the identification of a terpene synthase gene in wheat. It encodes a sesquiterpene synthase that catalyzes &beta, patchoulene formation with E,E-farnesyl diphosphate (FPP) as the substrate, thus named as TaPS. TaPS exhibits inducible expression in wheat in response to various elicitations. Particularly, alamethicin treatment strongly induces TaPS gene expression and &beta, patchoulene accumulation in wheat. Overexpression of TaPS in Arabidopsis successfully produces &beta, patchoulene, verifying the biochemical function of TaPS in planta. Furthermore, these transgenic Arabidopsis plants exhibit resistance against herbivory by repelling beet armyworm larvae feeding, thereby indicating anti-herbivory activity of &beta, patchoulene. The catalytic mechanism of TaPS is also explored by homology modeling and site-directed mutagenesis. Two key amino acids are identified to act in protonation and stability of intermediates and product formation. Taken together, one wheat sesquiterpene synthase is identified as &beta, patchoulene synthase. TaPS exhibits inducible gene expression and the sesquiterpene &beta, patchoulene is involved in repelling insect infestation.

A major and stably expressed QTL for spikelet number per spike identified in a 2-cM interval on chromosome arm 2DS was validated using two populations with different genetic backgrounds. Spikelet number per spike (SNS) plays a key role in wheat yield improvement. Numerous genetic and environmental factors influencing SNS have been documented, but the number of major, stably expressed and validated loci underlying SNS is still limited. In this study, a recombinant inbred line (RIL) population derived from a normal spikelet cultivar and a multiple-spikelet wheat line (with a longer spike with more canonically oriented apical spikelets) was genotyped using a Wheat55K single-nucleotide polymorphism (SNP) array and simple sequence repeat (SSR) markers. SNS was measured for this RIL population in eight environments. Five QTL were each identified in two or more environments. One of them, QSns.sau-2D (LOD = 3.47–38.24, PVE = 10.16–45.68%), was detected in all the eight environments. The QTL was located in a 2-cM interval on chromosome arm 2DS flanked by the markers AX-109836946 and AX-111956072. This QTL, QSns.sau-2D, significantly increased SNS by up to 14.72%. Several genes associated with plant growth and development were identified in the physical interval of QSns.sau-2D. This QTL was further validated by the tightly linked Kompetitive Allele Specific PCR (KASP) marker, KASP-AX-94721936, in two other populations with different genetic backgrounds. The significant correlation between SNS and anthesis date, plant height, spike length, grain number per spike and thousand-grain weight were detected and discussed. These results lay the foundation for fine mapping and cloning gene(s) underlying QSns.sau-2D.

Phosphoglucan phosphatases (Like-SEX4 1 and 2; LSF1 and LSF2) were reported to play roles in starch metabolism in leaves of Arabidopsis. In this study, we identified and mapped the LSF1 and LSF2 genes in barley (HvLSF1 and HvLSF2), characterized their gene and protein structures, predicted the cis-elements of their promoters, and analysed their expression patterns. HvLSF1 and HvLSF2 were mapped on the long arm of chromosome 1H (1HL) and 5H (5HL), respectively. Our results revealed varied exon-intron structures and conserved exon-intron junctions in both LSF1 and LSF2 from a range of analysed species. Alignment of protein sequences indicated that cTP and CT domains are much less varied than the functional domains (PDZ, DPS and CBM48). LSF2 was mainly expressed in anthers of barley and rice, and in leaf of Arabidopsis. LSF1 was mainly expressed in endosperm of barley and leaf of Arabidopsis and rice. The expression of LSF1 exhibited a diurnal pattern in rice only and that of LSF2 in both rice and Arabidopsis. Of the investigated stresses, only cold stress significantly reduced expression level of LSF1 and LSF2 in barley and LSF2 in Arabidopsis at late stages of the treatments. While heat treatment significantly decreased expression levels of LSF1 at middle stage (4 h) of a treatment in Arabidopsis only. The strong relationships detected between LSF2 and starch excess4 (SEX4), glucan, water dikinases or phosphoglucan, water dikinases were identified and discussed. Taken together, these results provide information of genetic manipulation of LSF1 and LSF2, especially in monocotyledon and further elucidate their regulatory mechanism in plant development.

An EMS-induced single-base mutation at a splice site caused abnormal RNA splicing and resulted in the gene inactivation and the lack of Wx-A1 protein in a wheat EMS mutant line. An EMS-mutagenized population was generated using common wheat cv. SM126 consisting of 10,600 M2 plants. One Wx-A1 null mutant was identified through analyses of 390 grains produced from 130 M2 plants using electrophoresis analyses. The Wx-A1 sequences of parental line SM126 and M2-31 mutant were determined as 2781 bp, and there was only one SNP mutation between them. The SNP was a mutation from G to A at nucleotide sequence position 2168 bp (G2168A) downstream of the start codon which was located at the splicing site within the eighth intron. All 52 cDNA transcripts were found to be incorrectly spliced and can be summarized as five types of variations. The deletion of the exon and the exclusion of intron were structural features in abnormal splicing RNA. Together with the prediction of potential splice regulatory motifs, the mutation G2168A happened within the 5′ splice site of the eighth intron and destroyed the splice donor site from GU to AU, which may have brought about a barrier against correct RNA splice, and generated abnormal mRNA, which was the mechanism of the inactivation of Wx-A1 in M2-31. The lack of Wx-A1 has resulted in changes in starch properties in the M2-31 mutant, with the reduction in amylose and starch contents. The increased grains hardness was observed in M2-31, which may be related to the lower expression level of Pinb-D1 gene. As the waxy wheat foods have a lot of advantages, the null waxy genes will be widely applied in breeding waxy wheat for varied amylose contents.

Chromosome translocation is an important driving force in shaping genomes during evolution. Detailed knowledge of chromosome translocations in a given species and its close relatives should increase the efficiency and precision of chromosome engineering in crop improvement. To identify genes flanking the breakpoints of translocations and inversions as a step toward identifying breakpoints in bread wheat, we systematically analysed genes in the Brachypodium genome against wheat survey sequences and bin-mapped ESTs (expressed sequence tags) derived from the hexaploid wheat genotype ‘Chinese Spring’. In addition to those well-known translocations between group 4, 5, and 7 chromosomes, this analysis identified genes flanking the three pericentric inversions on chromosomes 2B, 4B, and 5A. However, numerous chromosomal rearrangements reported in early studies could not be confirmed. The genes flanking the breakpoints reported in this study are valuable for isolating these breakpoints.

Waxy gene plays a key role for amylose synthesis in wheat seeds. The study evaluated the Waxy gene variability in 59 accessions of six diploid Triticum and Aegilops species. The percentages of variable sites, singleton variable sites, and parsimony informative sites were 16.38, 5.59, and 10.79 %, respectively. A total of 22 amino acid changes in the transit peptide and the remaining in the mature protein were observed. Moreover, 17 amino acid changes between Triticum and Aegilops species were also detected. Specially, Gla 14 and Phe 153 was observed in diploid Triticum, compared with Thr 14 and Tyr 153 in diploid Aegilops. Two types of amino acids, Gla 5/Val 5 and Ile 140/Val 140, were identified in T. urartu, as well as Val 22/Phe 22, Thr 52/Lys 52, and Gln 54/delete in A. tauschii. The insertion/deletions (InDels) had high frequency in intron region, but very low in transit peptide and exon region. Neighbour-joining tree showed that 146 sequences from 23 species could be clustered into eight groups with the species characterizations. The Wx-B1 of polyploid Triticum were grouped with A. sharonensis, A. longissima, A. searsii, A. speltoides, while Wx-D1 of T. aestivum and T. spelta together with A. umbellulata, A. markgrafii, A. comosa, and A. tauschii. The Wx-A1 of polyploid Triticum were separated clearly with the diploid species. The Wx-B1 could be divided into two subgroups and maybe had two phylogenetic origins, but most of them were related to A. speltoides. Waxy gene of A. tauschii also had two subgroups, and the sequences from southern Caspian (Mazandaran, Iran) were more closed with Wx-D1. The variability of 5′ Un-Translation Region (UTR) of waxy was stronger than intron and exon region based on genetic distance. Phylogeography analysis showed that geography affected strongly the distribution of all accessions along the north–south axis based on the partial open reading fragment (ORF) of waxy gene, A. speltoides and A. longissima along the west–east axis and north–south axis based on exon, and A. tauschii along the west–east axis based on 5′ UTR, respectively. Our results suggest that diploid Triticum and Aegilops have new waxy gene resources; waxy gene could play a more important role in genetic exploitation, genetic relationship evaluation and phylogeography investigation.

Spike length (SL) is one of the most important components of spike morphology, and the uppermost internode represents an ideal organ to study the transportation system. We performed conditional and unconditional quantitative trait locus (QTL) mapping in two unrelated recombinant inbred line populations to precisely detect QTLs for uppermost internode length (UIL) and SL, and to dissect the genetic relationship between these two factors with plant height (PH). Both of the populations were derived from crosses with synthetic wheat. Ten repetitive QTLs for UIL and six environment-independent QTLs for SL were identified in this study, and twelve of these were completely independent of PH. Conditional QTL mapping analysis indicated that SL was more independent to PH than UIL was. The results indicated that the conditional QTL mapping method could evaluate PH component effects on PH, and thus accelerate the selection of suitable loci that improve commercial wheat morphology yet avoid changes to PH.

In this study, the ARF genes were cloned, sequenced and located on the chromosomes. The gene expression of various stress conditions were analyzed through RT-PCR. Two important features of ARF in wheat were found: (1) High sequences homology among species in mammalian and plant and (2) Four exons and three introns were conserved in Poaceae. In this study the coding genes of ADP-ribosylation Factors (ARF) were characterized and they were located on chromosomes 3AL and 2DL in common wheat and its diploid progenitors. Forty-seven candidate SNPs in ARF were detected which were located in exons (17 SNPs) and introns (30 SNPs), respectively. As expected, most of the SNPs (66.34%) in ARF were transitions and the rest (33.66%) were transversions. The expression difference of ARF under various environmental stresses (low-temperature, Abscisic Acid (ABA), Polyethylene Glycol (PEG), NaCl, stripe rust), in two stages (seedling and maturity) and in different tissues (root, stem, flag leaf and immature embryo) of 15 days post-flowering were investigated. The results revealed that the expression levels of ARF were affected by environmental stresses. PEG stress induced the highest level of ARF expression, followed by the stripe rust and ABA stresses.

Bioactive compounds such as phenols and phytic acid in wheat contribute to antioxidant capacities. (1) Background: Prior studies drew a general conclusion that the environment affected bioactive compounds greatly, but how the single environmental factor affects these characteristics remains unclear. (2) Methods: We conducted that twenty-eight winter wheat genotypes were grown in replicated trials at seven locations in China for two consecutive years and subdivided the environmental factor into five soil factors and six meteorological factors to evaluate the impact on the antioxidant capabilities and bioactive compounds contents of wheat grains by using principal component analysis (PCA). RT-PCR was used to identify gene expression of bioactive compounds under different conditions. (3) Results: Temperature affects bioactive compounds contents and antioxidant capacities greatly in wheat grains. Accumulation time, daylight length, and daily maximum temperature showed a high correlation with bioactive compounds contents and antioxidant capacities, especially in the vegetative growth phase. The gene TaMIPs related to phytic acid and TaPAL1, TaC3H1, TaC4H, Ta4CL1, and TaCOMT1 related to total phenolics had higher gene expression level with larger temperature differences in wheat grains. (4) Conclusions: The planting locations with higher temperatures and longer daylight length could produce higher contents of bioactive compounds and antioxidant capacities and the cooler temperatures of a planting location might produce wheat grains with lower phytic acid contents in wheat grains.

Stripe rust (or yellow rust), caused by the fungus Puccinia striiformis f. sp. tritici (Pst), is one of the most important foliar diseases of wheat. Characterization and utilization of novel resistant genes is the most effective, economic and environmentally friendly approach to controlling the disease. Wheat line LM168-1, which was derived from a cross between common wheat Chuannong 16 and Milan, has good adult-plant resistance to stripe rust, based on field tests over several years. To elucidate the genetic basis of resistance, LM168-1 was crossed with susceptible variety SY95-71. Parents and F1, F2, BC1 and F2:3 progenies were tested in 2009–2011 in a field inoculated with the predominant races of Pst in China. The genetic analysis showed that resistance to stripe rust in LM168-1 was controlled by a single recessive gene, temporarily designated yrLM168. Simple sequence repeat (SSR), resistance gene analog polymorphism (RGAP) and target region amplification polymorphism (TRAP) techniques were used to identify molecular markers linked to the resistance locus. Finally, a linkage group consisting of two SSR, four RGAP and five TRAP markers was constructed for yrLM168 with 102 F2 plants. The closest markers R1 and R2 flanked the resistance gene locus at 2.4 and 2.4 cM, respectively. Furthermore, two SSR markers Xwmc59 and Xwmc145 assigned the gene to chromosome 6A. Because yrLM168 confers high-level resistance to the predominant races of Pst in China, it should be useful in stripe rust resistance breeding programs. The closely linked markers can be used for rapidly transferring yrLM168 to wheat breeding populations.

Gene loss during the formation of hexaploid bread wheat has been repeatedly reported. However, our knowledge on genome-wide analysis of the genes present on a single subgenome (SSG) in bread wheat is still limited. In this study, by analysing the ‘Chinese Spring’ chromosome arm shotgun sequences together with high-confidence gene models, we detected 433 genes on a SSG. Greater gene loss was observed in A and D subgenomes compared with B subgenome. More than 79% of the orthologs for these SSG genes were detected in diploid and tetraploid relatives of hexaploid wheat. Unexpectedly, no bias in expression breadth or in the distribution patterns of GO (gene ontology) terms for these genes was detected among the high-confidence genes. Further, network and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analyses indicated that most of these genes were not functionally related to each other. Interestingly, 30.7% of these SSG genes were most highly expressed in root, showing biased distribution given the distribution of the whole high-confidence genes. Collectively, these results facilitate our understanding of the loss of the genes that were retained in a SSG during the formation of hexaploid wheat.

Pre-harvest sprouting (PHS) causes significant yield loss and degrade the end-use quality of wheat, especially in regions with prolonged wet weather during the harvesting season. Unfortunately, the gene pool of Triticum durum (tetraploid durum wheat) has narrow genetic base for PHS resistance. Therefore, finding out new genetic resources from other wheat species to develop PHS resistance in durum wheat is of importance. A major PHS resistance QTL, Qphs.sicau-3B.1, was mapped on chromosome 3BL in a recombinant inbred line population derived from ‘CSCR6’ (Triticum spelta), a PHS resistant hexaploid wheat and ‘Lang’, a PHS susceptible Australian hexaploid wheat cultivar. This QTL, Qphs.sicau-3B.1, is positioned between DArT marker wPt-3107 and wPt-6785. Two SCAR markers (Ph3B.1 and Ph3B.2) were developed to track this major QTL and were used to assay a BC2F8 tetraploid population derived from a cross between the durum wheat ‘Bellaroi’ (PHS susceptible) and ‘CSCR6’ (PHS resistant). Phenotypic assay and marker-assisted selection revealed five stable tetraploid lines were highly PHS resistant. This study has successfully established that PHS-resistance QTL from hexaploid wheat could be efficiently introgressed into tetraploid durum wheat. This tetraploid wheat germplasm could be useful in developing PHS resistant durum cultivars with higher yield and good end-use quality.

A meta-analysis of quantitative trait loci (QTL) associated with tolerance to abiotic stresses in barley was carried out using results from 35 different experiments. “MetaQTL” software was used to project QTL positions on a reference map. Three hundred and thirty-seven QTL for traits associated with tolerance to abiotic stresses were included in this analysis which identified 79 metaQTL (MQTL) including 26 for drought, 11 for low temperature, 22 for salinity, 17 for water-logging, and 3 for mineral toxicity and deficiency. The distribution of MQTL was similar to that of the initial QTL. Many of these MQTL were located on chromosomes 2H (mainly for water-logging and drought) and 5H (mainly for salinity and low temperature). It inferred that chromosomes 2H and 5H were important for researches on barley abiotic tolerance, and the genes associated with abiotic stresses were concentrated relatively. As expected from trait correlations, 22.8 % of these MQTL displayed overlapping CIs. These overlapping regions were mainly on chromosomes 1H, 2H and 4H. The results indicated that the tolerance to diverse abiotic stresses were associated with each other in barley. Additionally, 67 candidate genes responsive to abiotic stresses were co-located with the abiotic-stress MQTL. Of them, a total of 55 had different conserved motifs. It inferred that the tolerance to abiotic stresses was contributed by multi-genes with diverse functions, though there might be some important genes associated with the tolerance to abiotic stresses in barley. Additionally, of these candidate genes, scsnp02622, scsnp01644 and scsnp19641 could be better for further studies of abiotic stresses tolerance.

5′ Untransalted regions (UTR) sequences of Waxy genes were amplified from all 81 Triticum L. and Aegilops L. species by PCR with specific primers. It was found that the sequence length at 7D loci was longer than that 7B and 7A. These sequences contained 170 singleton variable sites and 484 polymorphic sites and that the average length of Indels was 8.5 bp. There were abundant regions of restriction enzyme sites and two regions of simple sequence repeat, “GAA” and “CTGA”, in all sequences. A total of 65 uORFs were detected and classed into 37 types, with the variation in uORFs mainly due to single nucleotide polymorphisms (SNPs), and also to the presence of Indels. All sequences in tetraploids and hexaploids could be grouped into Types A, B, I, II, GI and GII based on sequence variation. Restriction enzyme sites, Indel polymorphisms and the classes of uORFs present together indicated that Type I was more similar to Ae. tauschii whereas Type II was more similar to Type B, and both more similar to Ae. longissima; Type A was more similar to Ae. speltoides. Population analysis was performed and Neighbour-joining trees derived from different species, types and accessions further confirmed that the ancestors of T. urartu, Ae. speltoides, Ae. longissima and Ae. tauschii were involved to the evolution of common wheat, and also implied that Ae. longissima might have participated later than Ae. speltoides. 5′ UTRs of Waxy genes in tetraploid and hexaploid species conserved characters from their respective progenitors when compared with diploid. For the first time, we are able to conclude there is abundant variation in SNPs, Indels and uORFs between 5′ UTRs of Waxy genes from different species related to common wheat, and suggest that further research could help to understand Waxy gene function more deeply and hence improve wheat breeding. Our results also show that three hexaploid species in China have unique diversity in the 5′ UTR of their Waxy gene.

Six hundred and thirty gene sequences from 21 different genomes in Triticeae tribe were obtained and subjected to phylogenetic analysis. The sequences showed high homology in both nucleotide sequences and length variation, and had a common conserved cysteine skeleton C–Xn–C–Xn–C–Xn–CC–Xn–C–X–C–Xn–C–Xn–C–Xn–C. The sequences from common wheat formed three clusters; two were close to Aegilops tauschii and Aegilops speltoides sequences, respectively, and the third cluster was complex with sequences from Ae. speltoides, Aegilops searsii, and Aegilops bicornis. Different S genome(s) of Aegilops contributed α-amylase inhibitor loci to polyploid wheat by gene introgression in interspecific hybridizations. No sequence from common wheat was similar to that from einkorn wheat. We conclude that the occurrence of multiple chromosomal translocations or inversions in the different genomes of Triticeae had not dramatically affected the primary structure of dimeric α-amylase inhibitors. The results revealed important information on genome shaping events and processes occurring at the dimeric α-amylase inhibitor genes loci and their bearing on the phylogenetic relationships in the tribe Triticeae (Poaceae).

High molecular weight (HMW) glutenin subunits (GS) play a key role in the determination of end-use quality of wheat and other cereal crops. In this study, we report the isolation and characterization of both promoter region and ORF of novel HMW-GS allele 1St1.3 from a perennial Triticeae species, Elymus canadensis. The amino acid (AA) sequences of E. canadensis 1St1.3 were deduced as 434 aa. Its protein primary structure comprises a signal peptide with a conserved N-terminal domain, a central repetitive domain and a C-terminal domain. E. canadensis 1St 1.3 possesses several distinct characteristics which are different from those of wheat HMW-GSs. The N-terminal domains of E. canadensis 1St 1.3 resemble that of y-type subunits, while their C-terminal domains are more similar to x-type subunits. The deletion of 85 bp fragment has been observed in promoter region of 1St 1.3, however which has not interrupted the expression of this gene. Our results indicate that 1St 1.3 is novel HMW-GS variants which will be valuable for enhancing our understanding of structural differentiation and the evolutionary relationship among HMW-GSs in Triticeae species.

High molecular weight (HMW) glutenin subunits (GS) are important seed storage proteins relevant to the end-use quality of wheat and other cereal crops. Here we report the isolation and characterization of two novel HMW-GS alleles (1St 1.4 and 1St1.1) from the perennial Triticeae species Elymus glaucus. The amino acid (aa) sequences of E. glaucus 1St1.4 and 1St1.1 were predicted as 434 aa and 358 aa, respectively. Both subunits comprise a signal peptide with a conserved N-terminal domain, a central repetitive domain and a C-terminal domain. Elymus glaucus 1St 1.4 and 1St1.1 exhibit several distinct characteristics different from other known HMW-GSs. The lengths of repetitive domains in E. glaucus 1St 1.4 and 1St1.1 are substantially smaller than those of other known HMW-GSs, in which 1St1.1 (only 358 aa) is the smallest subunit identified so far. The N-terminal domains of E. glaucus 1St 1.4 and 1St1.1 are homologous to y-type subunits, whereas their C-terminal domains are similar to x-type subunits. Our results indicate that E. glaucus 1St 1.4 and 1St1.1 are novel HMW-GS variants or isoforms, and the characterization of both subunits can enhance our understanding on the structural differentiation and evolutionary relationship of HMW-GSs in Triticeae species.

With 3 figures and 3 tables Abstract Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most devastating diseases in common wheat (Triticum aestivum L.). With the objective of identifying and tagging a new gene for resistance to stripe rust in wheat line P81, F1, F2 and F2:3 populations from the cross ‘Chuanmai 28’/P81 were inoculated with Chinese PST race CYR32 in greenhouse and field trials. P81 carried a single dominant gene for resistance (designated YrP81) to CYR32. Tests of allelism showed that YrP81 was different from Yr5, Yr10, Yr15 and Yr26. Simple sequence repeat (SSR) and resistance gene-analogue polymorphism (RGAP) between the parents were used for genotyping the F2 populations. YrP81 was closely linked to four SSR loci on chromosome 2BS with genetic distances of 18.3 cM (Xwmc25), 1.8 cM (Xgwm429), 4.1 cM (Xwmc770) and 5.3 cM (Xgwm148). Two RGAP markers RGA1 (NLRR/XLRR) and RGA2 (Pto kin4/NLRR-INV2) were also closely linked to YrP81 with genetic distances of 4.7 and 6.3 cM, respectively. The linkage map of YrP81 and molecular markers was established in the order Xwmc25-RGA2-RGA1-Xgwm429-YrP81-Xwmc770-Xgwm148. Pedigree analysis, response patterns with Chinese PST races and associations with markers suggested that YrP81 is a novel stripe rust resistance gene. The PCR-based microsatellite and RGAP markers identified here could be applied in selection of YrP81 in wheat breeding.

Yellow or stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a devastating foliar disease that affects common wheat (Triticum aestivum L.) around the world. In China, common wheat landraces are potential sources of disease and abiotic stress resistance genes for wheat improvement. Yilongtuomai (YL), a wheat landrace from Yilong County, Sichuan Province, shows high levels of resistance against most Chinese Pst races. In this study, the resistance of YL to stripe rust disease was examined in detail. Parent strains, YL and Taichung 29, a variety susceptible to Pst race CYR32, and their F1, F2, and F2:3 offspring, were inoculated with CYR32 during the seedling stage in the field or adult-plant stage in the greenhouse. Results indicated that resistance to CYR32 in YL is conferred by a single dominant gene, designated YrYL The segregating F2 population (352 plants), was analyzed in terms of its resistance locus using simple sequence repeats (SSRs), resistance gene analog polymorphisms (RGAPs), and sequence-related amplified polymorphism (SRAP). A linkage group of 6 SSRs, 2 RGAPs, and 1 SRAP was constructed for the YrYL gene. Using the identified SSRs associated with physical mapping of RGAP using Chinese Spring nullisomic-tetrasomic stocks, the YrYL gene was localized to the short arm of chromosome 7D. The gene was flanked by 1 SSR marker, Xbarc92, and 1 RGAP marker, CLRRfor/Ptokin4, at genetic distances of 5.35 and 9.86 cM, respectively. The YrYL gene was compared to other stripe rust resistance genes reported on chromosome 7D by evaluating its reaction patterns to CYR32 and its pedigree relationship. Our results suggest that the YrYL gene is a new stripe rust resistance gene.

Fusarium graminearum is the major causal agent of Fusarium head blight (FHB) of wheat and barley and is considered to be one of the most devastating plant diseases worldwide. Chitin is a critical component of the fungal cell wall and is polymerized from UDP-N-acetyl-alpha-D-glucosamine by chitin synthase. We characterized FgCHS8, a new class of the chitin synthase gene in F. graminearum. Disruption of FgCHS8 resulted in reduced accumulation of chitin, decreased chitin synthase activity, and had no effect on conidia growth when compared with the wild-type isolate. ΔFgCHS8 had a growth rate comparable to that of the wild-type isolate in vitro. However, ΔFgCHS8 had reduced growth when grown on agar supplemented with either 0.025% SDS or 0.9 mM salicylic acid. ΔFgCHS8 produced significantly less deoxynivalenol and exhibited reduced pathogenicity in wheat spikes. Re-introduction of a functional FgCHS8 gene into the ΔFgCHS8 mutant strain restored the wild-type phenotypes. Fluorescence microscopy revealed that FgCHS8 protein was initially expressed in the septa zone, and then gradually distributed over the entire cellular membrane, indicating that FgCHS8 was required for cell wall development. Our results demonstrated that FgCHS8 is important for cell wall sensitivity to environmental stress factors and deoxynivalenol production in F. graminearum.

The identification and characterization of resistance genes effective against stripe rust of wheat is beneficial for modern wheat breeding programs. Molecular markers to such genes facilitate their deployment. The variety Milan has resistance that is effective against the predominant stripe rust races in the Sichuan region. Two resistant and two susceptible F8 lines from a cross between Milan and the susceptible variety Chuannong 16 were used to investigate inheritance of the Milan resistance. Three F2 populations were developed from crosses between the resistant lines and their susceptible sibling lines (LM168a × LM168c, LM168c × LM168a, LM168b × LM168d) and used for genetic analysis and molecular mapping of the genes for resistance. The stripe rust resistance in LM168a and LM168b was conferred by a single dominant gene, temporarily designated as YrLM168a. Forty-five extreme susceptible plants from the F2 families of LM168d × LM168b were genotyped with 836 simple sequence repeat (SSR) markers to map YrLM168a. YrLM168a was mapped in chromosome 6BL. The nearest flanking markers Xwmc756 and Xbarc146 were 4.6 and 4.6 cM away from the gene at both sides, respectively. The amplification results of twenty extreme resistant (IT 0) and susceptible (IT 4) F2 plants of LM168c × LM168a and LM168a × LM168c with marker Xwmc756 further validated the mapping results. The study suggested that extreme individuals and recessive phenotype class can be successfully used for mapping genes, which should be efficient and reliable. In addition, the flanking markers near YrLM168a should be helpful in marker-assisted breeding.

The function of starch phosphorylase has long been debated on the regulation of starch metabolism during the growth and development of plants. In this study, we isolated starch phosphorylase genes (Pho1 and Pho2) from barley, characterized their gene and protein structures, predicated their promoter’s cis-elements and analyzed expression patterns. Multiple alignments of these genes showed that (1) both Pho1 and Pho2 genes possess 15 exons and 14 introns in all but three of the species analyzed, Aegilops tauschii (for Pho1 which contains 16 exons and 15 introns), potato (for Pho1b which contains 14 exons and 13 introns), and Triticum uraru (for Pho2 which contains 15 exons and 14 introns); (2) the exon–intron junctions of Pho1 and Pho2 flanking the ligand-binding sites are more conservative than the other regions. Analysis of protein sequences revealed that Pho1 and Pho2 were highly homologous except for two regions, the N terminal domain and the L78 insertion region. The results of real-time quantitative PCR (RT-qPCR) indicated that Pho2 is mainly expressed in germinating seeds, and the expression of Pho1 is similar to that of starch synthesis genes during seed development in barley. Microarray-based analysis indicated that the accumulation of Pho1 or Pho2 transcripts exhibited uniform pattern both in various tissues and various stages of seed development among species of barley, rice, and Arabidopsis. Pho1 of barley was significantly down-regulated under cold and drought treatments, and up-regulated under stem rust infection. Pho2 exhibited similar expression to Pho1 in barley. However, significant difference in expression was not detected for either Pho1 or Pho2 under any of the investigated abiotic stresses. In Arabidopsis, significant down-regulation was detected for Pho1 (PHS1) under abscisic acid (ABA) and for Pho2 (PHS2) under cold, salt, and ABA. Our results provide valuable information to genetically manipulate phosphorylase genes and to further elucidate their regulatory mechanism in the starch biosynthetic pathway.

Flavonoids are plant secondary metabolites that contribute to the adaptation of plants to environmental stresses, including resistance to abiotic and biotic stress. Flavonoids are also beneficial for human health and depress the progression of some chronic diseases. The biosynthesis of flavonoids, which belong to a large family of phenolic compounds, is a complex metabolic process with many pathways that produce different metabolites, controlled by key enzymes. There is limited knowledge about the composition, biosynthesis and regulation of flavonoids in cereals. Improved understanding of the accumulation of flavonoids in cereal grains would help to improve human nutrition through these staple foods. The biosynthesis of flavonoids, scope for altering the flavonoid composition in cereal crops and benefits for human nutrition are reviewed here.

Genetic variations of the four subgroups of m-type low-molecular-weight glutenin subunits (LMW-GS) genes and their associations with dough quality parameters were investigated through the groupspecific markers in 68 landraces of Triticum turgidum ssp. turgidum from China. The number of alleles were two, three, three and two respectively by the markers Glu3A.2, Glu3B.1, Glu3B.2 and Glu3A.3, which was one-to-one correspondence to the “MDTSCIP-”, “MENSHIP-”, “METSHIPS-” and “METSCIP-” subgroups of m-type LMW-GS genes respectively. Results from the sequencing of PCR products further confirmed m-type LMW-GS genes from tetraploid landraces. The widespread variation of dough quality parameters were observed in all accessions and their association with the alleles of m-type LMW-GS genes were also investigated by crosstab analysis. We found that allele variations in “METSHIPS-” subgroup of m-type LMW-GS genes were significantly linked with gluten index at (K=14.02, P

Eight putative barley AP2/EREBP genes were cloned based on the barley HarvEST database. The introns in the translated regions of these genes were all located at the front of the conserved AP2 domain. Among the eight genes, DNA sequences of hv.18885 and hv.17070 were the same between Steptoe and Morex. Of the other six genes, there was very high ratio of ‘G’ and ‘C’ bases in the variant DNA sequence regions of three genes between Steptoe and Morex. Thus, only three genes were successfully located on the chromosomes. hv.8737 and hv.15732 were mapped to the short arm of chromosome 7H and hv.2871 to a major quantitative trait loci (QTL) region conferring adult spot blotch tolerance on the short arm of chromosomes 3H. Phylogenetic analysis showed that putative plant AP2/EREBP genes in the AP2 subfamily were completely separated from those in other subfamilies. The putative AP2/EREBP genes in RAV and Soloist subfamilies were clustered together, while those in the ERF and CRT/DRE subfamily were clustered together. Among the main branch-groups of each subfamily in plant, those from wheat, barley and rice had the tendency to cluster together when compared with those from Arabidopsis. Key words: AP2/EREBP, clone, map, phylogenetic, barley.

Pathogens are a reason for low yield in common wheat (Triticum aestivum L.). Stripe rust (or yellow rust), caused by the fungus Puccinia striiformis f. sp. tritici (Pst), is one of the most important foliar diseases of wheat. One of the most cost-effective and environmentally sound ways to control stripe rust is to use plant varieties that are resistant to this pathogen. It is an important task for wheat breeders and pathologists to identify new genes and pyramid them in order to achieve high-level, durable resistance to stripe rust. One spring wheat germplasm, HRMSN-81, with resistance to the most dominant races in China, is identified from the CIMMYT breeding lines. To elucidate the genetic basis of its resistance, HRMSN-81 was crossed with susceptible wheat genotype Taichung 29. Seedlings of the parents were tested with Chinese Pst isolates CYR31, CYR32, and CYR33 under controlled greenhouse conditions, and adult plants of the parents and F1, F2, and F2:3 progeny were inoculated with the epidemic stripe rust mixed races, including CYR31, CYR32, and CYR33, in fields under natural infection. Genetic analysis showed that HRMSN-81 has a single dominant gene conferring all-stage resistance, temporarily designated as YrHRMSN-81. Resistance gene analogue polymorphism (RGAP), simple sequence repeat (SSR), target region amplified polymorphism (TRAP), and sequence-related amplified polymorphism (SRAP) techniques in combination with bulked segregant analysis (BSA) were used to identify molecular markers linked to the resistance gene. A linkage map consisting of six RGAP, two SSR, one TRAP, and two SRAP markers was constructed for YrHRMSN-81 using 148 F2 plants. The gene was mapped to chromosome arm 2DS by testing the complete set of nulli-tetrasomic lines and selected ditelosomic lines with two RGAP markers and was further confirmed by two chromosome-specific SSR markers. The results of gene characteristics and chromosome locations indicated that YrHRMSN-81 was probably a new stripe rust resistance gene. The two flanking markers Xwgp-180bp (93% polymorphism rate) and Xwmc453 (91% polymorphism rate) detected 100% polymorphism of the 56 tested wheat genotypes when they were used in combination. The identification of the gene YrHRMSN-81 and the determination of the flanking markers should be useful for rapidly transferring it in wheat breeding programs.