Wheat powdery mildew resistance gene Pm64 derived from wild emmer (Triticum turgidum var.dicoccoides) is tightly linked in repulsion with stripe rust resistance gene Yr5

Stripe rust and powdery mildew are both devastating diseases for durum and common wheat.Pyramiding of genes conferring resistance to one or more diseases in a single cultivar is an important breeding approach to provide broader spectra of resistances in wheat improvement. A new powdery mildew resistance gene originating from wild emmer(Triticum turgidum var.dicoccoides) backcrossed into common wheat(T. aestivum) line WE35 was identified. It conferred an intermediate level of resistance to Blumeria graminis f. sp. tritici isolate E09 at the seedling stage and a high level of resistance at the adult plant stage. Genetic analysis showed that the powdery mildew resistance in WE35 was controlled by a dominant gene designated Pm64. Bulked segregant analysis(BSA) and molecular mapping indicated that Pm64 was located in chromosome bin 2 BL4-0.50–0.89. Polymorphic markers were developed from the corresponding genomic regions of Chinese Spring wheat and wild emmer accession Zavitan to delimit Pm64 to a 0.55 cM genetic interval between markers WGGBH1364 and WGGBH612, corresponding to a 15 Mb genomic region on Chinese Spring and Zavitan 2 BL, respectively. The genetic linkage map of Pm64 is critical for fine mapping and cloning. Pm64 was completely linked in repulsion with stripe rust resistance gene Yr5. Analysis of a larger segregating population might identify a recombinant line with both genes as a valuable resource in breeding for resistance to powdery mildew and stripe rust.

Effects of high temperature on the microstructure and mechanical behavior of hard coal

Coal is a common porous sedimentary rock whose microstructure and mechanical behavior are often affected by high temperature.In order to deeply investigate the effect of high temperature on the structure and mechanical properties of coal,six groups of coal samples with different heating temperatures were prepared.Various testing methods,including ultrasonic test,scanning electron microscopy,nuclear magnetic resonance and uniaxial compression test,were used in this study.The results showed that the physical and chemical change processes of coal samples under high temperature can be divided into two stages:the stage of drying and degassing and the stage of pyrolysis of the coal matrix.With the increase of heating temperature,the total porosity of coal samples increases,while the integrity and the deformation resistance of coal samples decrease.In addition,the elastic modulus and uniaxial compressive strength of the coal sample decrease with the increase of heating temperature.The Poisson’s ratio decreases firstly and then increases with the increase of heating temperature.

Mapping stripe rust resistance genes by BSR-Seq:YrMM58 and YrHY1 on chromosome 2AS in Chinese wheat lines Mengmai 58 and Huaiyang 1 are Yr17

Stripe rust(yellow rust), caused by Puccinia striiformis f. sp. tritici(PST),is one of the most devastating fungal diseases in common wheat(Triticum aestivum L.) in China and worldwide. Resistance breeding is the most effective strategy to control diseases in crop plants. Chinese wheat lines Mengmai 58 and Huaiyang 1 are highly resistant to PST race CYR34(V26) at the adult plant stage. To genetically map the underlying resistance genes we developed segregating populations by crossing Mengmai 58 and Huaiyang 1 with the susceptible cultivar Nongda 399. The stripe rust resistances in Mengmai 58 and Huaiyang 1 were both controlled by single dominant genes, provisionally designated YrMM58 and YrHY1, respectively. Bulked segregant RNA-Seq(BSR-Seq) analysis showed that YrMM58 and YrHY1 were located in the same distal ~16 Mb region on chromosome 2 AS.Comparative genomics analysis with the physical map of Aegilops tauschii proved useful for developing additional markers to saturate the genetic linkage map. YrMM58 and YrHY1 were mapped to the distal end of chromosome arm 2 AS, with the closest marker WGGB148 being 7.7 cM and 3.8 cM from the resistance gene, which was considered to be Yr17. These markers can be used in marker-assisted selection.

Mapping a leaf senescence gene els1 by BSR-Seq in common wheat

Leaf senescence is normally the last stage of plant development. Early senescence of functional leaves significantly reduces the photosynthetic time and efficiency, seriously affecting grain yield and quality in wheat. Discovering genes responsible for early leaf senescence(els) are necessary for developing novel germplasms and cultivars with delayed leaf-senescence through molecular manipulation and marker assisted selection. In this study, we identified an early leaf senescence line M114 in a derivative of a wheat breeding population. Genetic analysis indicated that early leaf senescence in M114 is controlled by a single recessive gene, provisionally designated els1. By applying bulked segregant analysis and RNA-Seq(BSR-Seq), seven polymorphic markers linked to els1 were developed and the gene was located on chromosome arm 2 BS in a 1.5 c M genetic interval between markers WGGB303 and WGGB305. A co-segregating marker, WGGB302, provide a starting point for fine mapping and map-based cloning of els1.

Mapping of wheat stripe rust resistance gene Yr041133 by BSR-Seq analysis

Puccinia striiformis Westend. f. sp. tritici(Pst) pathotype CYR34 is widely virulent and prevalent in China.Here, we report identification of a strpie rust resistance(Yr) gene, designated Yr041133, in winter wheat line 041133. This line produced a hypersensitive reaction to CYR34 and conferred resistance to 13 other pathotypes. Resistance to CYR34 in line 041133 was controlled by a single dominant gene. Bulked segregant RNA sequencing(BSR-Seq) was performed on a pair of RNA bulks generated by pooling resistant and susceptible recombinant inbred lines. Yr041133 was mapped to a 1.7 c M genetic interval on the chromosome arm 7 BL that corresponded to a 0.8 Mb physical interval(608.9–609.7 Mb) in the Chinese Spring reference genome. Based on its unique physical location Yr041133 differred from the other Yr genes on this chromosome arm.

RNA-seq analysis of Brachypodium distachyon responses to Barley stripe mosaic virus infection

Barley stripe mosaic virus(BSMV) is the type member of the genus Hordeivirus. Brachypodium distachyon line Bd3-1 shows resistance to the BSMV ND18 strain, but is susceptible to an ND18 double mutant(βNDTGB1R390K, T392K) in which lysine is substituted for an arginine at position 390 and for threonine at position 392 of the triple gene block 1(TGB1) protein. In order to understand differences in gene expression following infection with ND18 and double mutant ND18, Bd3-1 seedlings were subjected to RNA-seq analyses at 1, 6, and14 days post inoculation(dpi). The results revealed that basal immunity genes involved in cellulose synthesis and pathogenesis-related protein biosynthesis were enhanced in incompatible interactions between Bd3-1 and ND18. Most of the differentially expressed transcripts are related to trehalose biosynthesis, ethylene, jasmonic acid metabolism,protein phosphorylation, protein ubiquitination, transcriptional regulation, and transport process, as well as pathogenesis-related protein biosynthesis. In compatible interactions between Bd3-1 and ND18 mutant, Bd3-1 developed weak basal resistance responses to the virus. Many genes involved in cellulose biosynthesis, protein amino acid phosphorylation,protein biosynthesis, protein glycosylation, glycolysis and cellular macromolecular complex assembly that may be related to virus replication, assembly and movement were up-regulated. Some genes involved in oxidative stress responses were also up-regulated at14 dpi. BSMV ND18 mutant infection suppressed expression of genes functioning in regulation of transcription, protein kinase, cellular nitrogen compound biosynthetic process and photosynthesis. Differential expression patterns between compatible and incompatible interactions in Bd3-1 to the two BSMV strains provide important clues for understanding mechanism of resistance to BMSV in the model plant Brachypodium.

A 36 Mb terminal deletion of chromosome 2BL is responsible for a wheat semi-dwarf mutation

Reduced plant height is one of the most important traits related to lodging resistance and crop yield. The use of reduced height genes has been one of the main features in breeding modern high-yielding wheat varieties with less lodging. A spontaneous dwarf mutant DD399 was identified in a high yielding, gibberellic acid(GA)-insensitive, lodging-resistant variety Nongda 399(ND399). Significant differences in upper internode lengths between mutant DD399 and wild type ND399 were caused by reduced cell elongation. The plant height of ND399 × DD399 F_(1) hybrids was intermediate between the parents, indicating incomplete dominance or a dose–response effect of a reduced height gene. Plant height showed continuous distribution in the F_(2) population, and segregation distortion was observed among the 2292 F_(2:3) progenies. The reduced height mutation was characterized by Illumina 90 K iSelect SNP genotyping and bulked segregant RNA-Seq(BSR-Seq) analysis of the segregating population. A concentrated cluster of polymorphic SNPs associated with the reduced height phenotype was detected in the distal region of chromosome arm 2 BL. Co-segregation of reduced height phenotype with the clustered markers revealed a 36 Mb terminal deletion of chromosome 2 BL in mutant DD399.

WPA1 encodes a vWA domain protein that regulates wheat plant architecture

Plant height,spike,leaf,stem and grain morphologies are key components of plant architecture and related to wheat yield.A wheat(Triticum aestivum L.)mutant,wpa1,displaying temperaturedependent pleiotropic developmental anomalies,was isolated.The WPA1 gene,encoding a von Willebrand factor type A(vWA)domain protein,was located on chromosome arm 7DS and isolated by map-based cloning.The functionality of WPA1 was validated by multiple independent EMS-induced mutants and gene editing.Phylogenetic analysis revealed that WPA1 is monocotyledon-specific in higher plants.The identification of WPA1 provides opportunity to study the temperature regulated wheat development and grain yield.

Utility of Triti-Map for bulk-segregated mapping of causal genes and regulatory elements in Triticeae

Triticeae species,including wheat,barley,and rye,are critical for global food security.Mapping agronomically important genes is crucial for elucidating molecular mechanisms and improving crops.However,Triticeae includes many wild relatives with desirable agronomic traits,and frequent introgressions occurred during Triticeae evolution and domestication.Thus,Triticeae genomes are generally large and complex,making the localization of genes or functional elements that control agronomic traits challenging.Here,we developed Triti-Map,which contains a suite of user-friendly computational packages specifically designed and optimized to overcome the obstacles of gene mapping in Triticeae,as well as a web interface integrating multi-omics data from Triticeae for the efficient mining of genes or functional elements that control particular traits.The Triti-Map pipeline accepts bothDNA and RNAbulk-segregated sequencing data as well as traditional QTL data as inputs for locating genes and elucidating their functions.We illustrate the usage of Triti-Map with a combination of bulk-segregated ChIP-seq data to detect a wheat disease-resistance gene with its promoter sequence that is absent from the reference genome and clarify its evolutionary process.We hope that Triti-Map will facilitate gene isolation and accelerate Triticeae breeding.

Functional characterization of powdery mildew resistance gene MIIW172,a new Pm60 allele and its allelic variation in wild emmer wheat

Wild emmer wheat(Triticum dicoccoides,WEW)is an immediate progenitor of both the cultivated tetraploid and hexaploid wheats and it harbors rich genetic diversity against powdery mildew caused by Blumeria graminis f.sp.tritici(Bgt).A powdery mildew resistance gene Ml I^(W172)originated from WEW accession I^(W172)(G-797-M)is fine mapped in a 0.048 centimorgan(c M)genetic interval on 7 AL,corresponding to a genomic region spanning 233 kb,1 Mb and 800 kb in Chinese Spring,WEW Zavitan,and T.urartu G1812,respectively.Ml I^(W172)encodes a typical NLR protein NLRI^(W172)and physically locates in an NBS-LRR gene cluster.NLRI^(W172)is subsequently identified as a new allele of Pm60,and its function is validated by EMS mutagenesis and transgenic complementation.Haplotype analysis of the Pm60 alleles reveals diversifications in sequence variation in the locus and presence and absence variations(PAV)in WEW populations.Four common single nucleotide variations(SNV)are detected between the Pm60 alleles from WEW and T.urartu,indicative of speciation divergence between the two different wheat progenitors.The newly identified Pm60 alleles and haplotypes in WEW are anticipated to be valuable for breeding powdery mildew resistance wheat cultivars via marker-assisted selection.