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Pacific Biosciences
January 1, 2019

Applying the latest advances in genomics and phenomics for trait discovery in polyploid wheat.

Improving traits in wheat has historically been challenging due to its large and polyploid genome, limited genetic diversity and in-field phenotyping constraints. However, within recent years many of these barriers have been lowered. The availability of a chromosome-level assembly of the wheat genome now facilitates a step-change in wheat genetics and provides a common platform for resources, including variation data, gene expression data and genetic markers. The development of sequenced mutant populations and gene-editing techniques now enables the rapid assessment of gene function in wheat directly. The ability to alter gene function in a targeted manner will unmask the effects…

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January 1, 2019

Rapid Gene Cloning in Wheat

The identification of wheat and barley genes controlling important agronomic traits using positional cloning has traditionally been a challenging and time-consuming procedure. This is due to the enormous genome size and high repeat content from transposable elements (TEs). Low marker density, suppressed recombination, and the high cost of generating a physical contig across a genetically defined map interval have further restricted the application of positional approximation. Over the past decade, the cost of DNA sequencing has significantly dropped, as has our ability to computationally analyze large quantities of DNA sequence data. This has enabled researchers to exploit next-generation sequencing (NGS)…

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August 1, 2018

Optical and physical mapping with local finishing enables megabase-scale resolution of agronomically important regions in the wheat genome.

Numerous scaffold-level sequences for wheat are now being released and, in this context, we report on a strategy for improving the overall assembly to a level comparable to that of the human genome.Using chromosome 7A of wheat as a model, sequence-finished megabase-scale sections of this chromosome were established by combining a new independent assembly using a bacterial artificial chromosome (BAC)-based physical map, BAC pool paired-end sequencing, chromosome-arm-specific mate-pair sequencing and Bionano optical mapping with the International Wheat Genome Sequencing Consortium RefSeq v1.0 sequence and its underlying raw data. The combined assembly results in 18 super-scaffolds across the chromosome. The value…

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June 1, 2018

Optimise wheat A-genome.

The wild einkorn wheat Triticum urartu (Tu) is the A-genome progenitor of tetraploid (AABB) and hexaploid (AABBDD) wheat. A draft genome of Tu was published in 2013, but a better reference sequence is urgently needed by scientists and breeders. Hong-Qing Ling, from the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and colleagues have now completed a high-quality Tu genome using multiple methods.

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May 1, 2018

Genome sequence of the progenitor of wheat A subgenome Triticum urartu.

Triticum urartu (diploid, AA) is the progenitor of the A subgenome of tetraploid (Triticum turgidum, AABB) and hexaploid (Triticum aestivum, AABBDD) wheat1,2. Genomic studies of T. urartu have been useful for investigating the structure, function and evolution of polyploid wheat genomes. Here we report the generation of a high-quality genome sequence of T. urartu by combining bacterial artificial chromosome (BAC)-by-BAC sequencing, single molecule real-time whole-genome shotgun sequencing 3 , linked reads and optical mapping4,5. We assembled seven chromosome-scale pseudomolecules and identified protein-coding genes, and we suggest a model for the evolution of T. urartu chromosomes. Comparative analyses with genomes of…

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March 26, 2018

Dynamic evolution of a-gliadin prolamin gene family in homeologous genomes of hexaploid wheat.

Wheat Gli-2 loci encode complex groups of a-gliadin prolamins that are important for breadmaking, but also major triggers of celiac disease (CD). Elucidation of a-gliadin evolution provides knowledge to produce wheat with better end-use properties and reduced immunogenic potential. The Gli-2 loci contain a large number of tandemly duplicated genes and highly repetitive DNA, making sequence assembly of their genomic regions challenging. Here, we constructed high-quality sequences spanning the three wheat homeologous a-gliadin loci by aligning PacBio-based sequence contigs with BioNano genome maps. A total of 47 a-gliadin genes were identified with only 26 encoding intact full-length protein products. Analyses…

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February 14, 2018

Development of molecular markers linked to powdery mildew resistance GenePm4bby combining SNP discovery from transcriptome sequencing data with bulked segregant analysis (BSR-Seq) in wheat.

Powdery mildew resistance genePm4b, originating fromTriticum persicum, is effective against the prevalentBlumeria graminisf. sp.tritici(Bgt) isolates from certain regions of wheat production in China. The lack of tightly linked molecular markers with the target gene prevents the precise identification ofPm4bduring the application of molecular marker-assisted selection (MAS). The strategy that combines the RNA-Seq technique and the bulked segregant analysis (BSR-Seq) was applied in an F2:3mapping population (237 families) derived from a pair of isogenic lines VPM1/7*Bainong 3217 F4(carryingPm4b) and Bainong 3217 to develop more closely linked molecular markers. RNA-Seq analysis of the two phenotypically contrasting RNA bulks prepared from the representative…

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February 1, 2018

Current advances in genome sequencing of common wheat and its ancestral species

Common wheat is an important and widely cultivated food crop throughout the world. Much progress has been made in regard to wheat genome sequencing in the last decade. Starting from the sequencing of single chromosomes/chromosome arms whole genome sequences of common wheat and its diploid and tetraploid ancestors have been decoded along with the development of sequencing and assembling technologies. In this review, we give a brief summary on international progress in wheat genome sequencing, and mainly focus on reviewing the effort and contributions made by Chinese scientists.

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January 30, 2018

Identification and expression analysis of wheat TaGF14 genes.

The 14-3-3 gene family members play key roles in various cellular processes. However, little is known about the numbers and roles of 14-3-3 genes in wheat. The aims of this study were to identify TaGF14 numbers in wheat by searching its whole genome through blast, to study the phylogenetic relationships with other plant species and to discuss the functions of TaGF14s. The results showed that common wheat harbored 20 TaGF14 genes, located on wheat chromosome groups 2, 3, 4, and 7. Out of them, eighteen TaGF14s are non-e proteins, and two wheat TaGF14 genes, TaGF14i and TaGF14f, are e proteins.…

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November 15, 2017

Genome sequence of the progenitor of the wheat D genome Aegilops tauschii.

Aegilops tauschii is the diploid progenitor of the D genome of hexaploid wheat (Triticum aestivum, genomes AABBDD) and an important genetic resource for wheat. The large size and highly repetitive nature of the Ae. tauschii genome has until now precluded the development of a reference-quality genome sequence. Here we use an array of advanced technologies, including ordered-clone genome sequencing, whole-genome shotgun sequencing, and BioNano optical genome mapping, to generate a reference-quality genome sequence for Ae. tauschii ssp. strangulata accession AL8/78, which is closely related to the wheat D genome. We show that compared to other sequenced plant genomes, including a…

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November 1, 2017

New insights into structural organization and gene duplication in a 1.75-Mb genomic region harboring the a-gliadin gene family in Aegilops tauschii, the source of wheat D genome.

Among the wheat prolamins important for its end-use traits, a-gliadins are the most abundant, and are also a major cause of food-related allergies and intolerances. Previous studies of various wheat species estimated that between 25 and 150 a-gliadin genes reside in the Gli-2 locus regions. To better understand the evolution of this complex gene family, the DNA sequence of a 1.75-Mb genomic region spanning the Gli-2 locus was analyzed in the diploid grass, Aegilops tauschii, the ancestral source of D genome in hexaploid bread wheat. Comparison with orthologous regions from rice, sorghum, and Brachypodium revealed rapid and dynamic changes only…

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October 23, 2017

The first near-complete assembly of the hexaploid bread wheat genome, Triticum aestivum.

Common bread wheat, Triticum aestivum, has one of the most complex genomes known to science, with 6 copies of each chromosome, enormous numbers of near-identical sequences scattered throughout, and an overall haploid size of more than 15 billion bases. Multiple past attempts to assemble the genome have produced assemblies that were well short of the estimated genome size. Here we report the first near-complete assembly of T. aestivum, using deep sequencing coverage from a combination of short Illumina reads and very long Pacific Biosciences reads. The final assembly contains 15 344 693 583 bases and has a weighted average (N50)…

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September 15, 2017

Overview of the wheat genetic transformation and breeding status in China.

In the past two decades, Chinese scientists have achieved significant progress on three aspects of wheat genetic transformation. First, the wheat transformation platform has been established and optimized to improve the transformation efficiency, shorten the time required from starting of transformation procedure to the fertile transgenic wheat plants obtained as well as to overcome the problem of genotype-dependent for wheat genetic transformation in wide range of wheat elite varieties. Second, with the help of many emerging techniques such as CRISPR/cas9 function of over 100 wheat genes has been investigated. Finally, modern technology has been combined with the traditional breeding technique…

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August 29, 2017

High-quality RNA isolation from wheat immature grains

Grain quality is one of the most important targets in wheat breeding. Transcriptome analyses of wheat developing grains and endosperm have been performed using the microarray and RNA sequencing (RNA-seq) approaches (Wan et al. 2008, 2009; Nemeth et al. 2010; Pellny et al. 2012; Dong et al. 2015). For the RNA-seq analysis of the grain transcriptome and precise quantification of each transcript in developing grain and endosperm, the high-quality RNA is essential. For the microarray analysis, =7.3 RIN (RNA integrity number) value for the RNA sample quality is required according to the Agilent microarray protocol. In the previous report for…

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August 10, 2017

Complete genome sequencing and targeted mutagenesis reveal virulence contributions of Tal2 and Tal4b of Xanthomonas translucens pv. undulosa ICMP11055 in bacterial leaf streak of wheat

Bacterial leaf streak caused by Xanthomonas translucens pv. undulosa (Xtu) is an important disease of wheat (Triticum aestivum) and barley (Hordeum vulgare) worldwide. Transcription activator-like effectors (TALEs) play determinative roles in many of the plant diseases caused by the different species and pathovars of Xanthomonas, but their role in this disease has not been characterized. ICMP11055 is a highly virulent Xtu strain from Iran. The aim of this study was to better understand genetic diversity of Xtu and to assess the role of TALEs in bacterial leaf streak of wheat by comparing the genome of this strain to the recently…

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