SummaryBread wheat (Triticum aestivum) is a globally important crop, accounting for 20% of the calories consumed by mankind. We sequenced its large and challenging 17 Gb hexaploid genome using 454 pyrosequencing and compared this with the sequences of diploid ancestral and progenitor genomes. Between 94,000-96,000 genes were identified, and two-thirds were assigned to the A, B and D genomes. High-resolution synteny maps identified many small disruptions to conserved gene order. We show the hexaploid genome is highly dynamic, with significant loss of gene family members upon polyploidization and domestication, and an abundance of gene fragments. Several classes of genes involved in energy harvesting, metabolism and growth are among expanded gene families that could be associated with crop productivity. Our analyses, coupled with the identification of extensive genetic variation, provide a new resource for accelerating gene discovery and improving this major crop.

BackgroundThe prevailing paradigm of host-parasite evolution is that arms races lead to increasing specialisation via genetic adaptation. Insect herbivores are no exception and the majority have evolved to colonise a small number of closely related host species. Remarkably, the green peach aphid, Myzus persicae, colonises plant species across 40 families and single M. persicae clonal lineages can colonise distantly related plants. This remarkable ability makes M. persicae a highly destructive pest of many important crop species.ResultsTo investigate the exceptional phenotypic plasticity of M. persicae, we sequenced the M. persicae genome and assessed how one clonal lineage responds to host plant species of different families. We show that genetically identical individuals are able to colonise distantly related host species through the differential regulation of genes belonging to aphid-expanded gene families. Multigene clusters collectively upregulate in single aphids within two days upon host switch. Furthermore, we demonstrate the functional significance of this rapid transcriptional change using RNA interference (RNAi)-mediated knock-down of genes belonging to the cathepsin B gene family. Knock-down of cathepsin B genes reduced aphid fitness, but only on the host that induced upregulation of these genes.ConclusionsPrevious research has focused on the role of genetic adaptation of parasites to their hosts. Here we show that the generalist aphid pest M. persicae is able to colonise diverse host plant species in the absence of genetic specialisation. This is achieved through rapid transcriptional plasticity of genes that have duplicated during aphid evolution.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-016-1145-3) contains supplementary material, which is available to authorized users.

SummaryTwo mutant lines of barley, Risù 17 and Notch-2, were found to accumulate phytoglycogen in the grain. Like the sugary mutants of maize and rice, these phytoglycogen-accumulating mutants of barley lack isoamylase activity in the developing endosperm. The mutants were shown to be allelic, and to have lesions in the isoamylase gene, isa1 that account for the absence of this enzyme. As well as causing a reduction in endosperm starch content, the mutations have a profound effect on the structure, number and timing of initiation of starch granules. There are no normal A-type or B-type granules in the mutants. The mutants have a greater number of starch granules per plastid than the wild-type and, particularly in Risù 17, this leads to the appearance of compound starch granules. These results suggest that, as well as suppressing phytoglycogen synthesis, isoamylase in the wild-type endosperm plays a role in determining the number, and hence the form, of starch granules.

SummaryFood security is a global concern and substantial yield increases in cereal crops are required to feed the growing world population. Wheat is one of the three most important crops for human and livestock feed. However, the complexity of the genome coupled with a decline in genetic diversity within modern elite cultivars has hindered the application of marker-assisted selection (MAS) in breeding programmes. A crucial step in the successful application of MAS in breeding programmes is the development of cheap and easy to use molecular markers, such as single-nucleotide polymorphisms. To mine selected elite wheat germplasm for intervarietal single-nucleotide polymorphisms, we have used expressed sequence tags derived from public sequencing programmes and next-generation sequencing of normalized wheat complementary DNA libraries, in combination with a novel sequence alignment and assembly approach. Here, we describe the development and validation of a panel of 1114 single-nucleotide polymorphisms in hexaploid bread wheat using competitive allele-specific polymerase chain reaction genotyping technology. We report the genotyping results of these markers on 23 wheat varieties, selected to represent a broad cross-section of wheat germplasm including a number of elite UK varieties. Finally, we show that, using relatively simple technology, it is possible to rapidly generate a linkage map containing several hundred single-nucleotide polymorphism markers in the doubled haploid mapping population of Avalon · Cadenza.

For more than 10,000 years, the selection of plant and animal traits that are better tailored for human use has shaped the development of civilizations. During this period, bread wheat (Triticum aestivum) emerged as one of the world's most important crops. We used exome sequencing of a world-wide panel of almost 500 genotypes selected from across the geographical range of the wheat species complex to explore how 10,000 years of hybridization, selection, adaptation and plant breeding shaped the genetic makeup of modern bread wheats. We observed considerable genetic variations at the genic, chromosomal and subgenomic levels deciphering the likely origins of modern day wheats, the consequences of range expansion and allelic variants selected since its domestication. Our data supports a reconciled model of wheat evolution and provides novel avenues for future breeding improvement.

The processes of quality assessment and control are an active area of research at The Genome Analysis Centre (TGAC). Unlike other sequencing centers that often concentrate on a certain species or technology, TGAC applies expertise in genomics and bioinformatics to a wide range of projects, often requiring bespoke wet lab and in silico workflows. TGAC is fortunate to have access to a diverse range of sequencing and analysis platforms, and we are at the forefront of investigations into library quality and sequence data assessment. We have developed and implemented a number of algorithms, tools, pipelines and packages to ascertain, store, and expose quality metrics across a number of next-generation sequencing platforms, allowing rapid and in-depth cross-platform Quality Control (QC) bioinformatics. In this review, we describe these tools as a vehicle for data-driven informatics, offering the potential to provide richer context for downstream analysis and to inform experimental design.

In higher plants several isoforms of starch synthase contribute to the extension of glucan chains in the synthesis of starch. Different isoforms are responsible for the synthesis of essentially linear amylose chains and branched, amylopectin chains. The activity of granule-bound starch synthase I from potato has been compared with that of starch synthase II from potato following expression of both isoforms in Escherichia coli. Significant differences in their activities are apparent which may be important in determining their specificities in vivo. These differences include affinities for ADPglucose and glucan substrates, activation by amylopectin, response to citrate, thermosensitivity and the processivity of glucan chain extension. To define regions of the isoforms determining these characteristic traits, chimeric proteins have been produced by expression in E. coli. These experiments reveal that the C-terminal region of granule-bound starch synthase I confers most of the specific properties of this isoform, except its processive elongation of glucan chains. This region of granule-bound starch synthase I is distinct from the C-terminal region of other starch synthases. The specific properties it confers may be important in defining the specificity of granule-bound starch synthase I in producing amylose in vivo.Keywords: amylopectin; amylose; starch synthase.In plants, starch synthases form starch by catalysing the transfer of the glucosyl moiety from ADPglucose to the nonreducing ends of glucan chains through an a1±4 linkage. Starch is composed of two types of glucan polymer: amylose, a predominantly linear a1±4-linked glucan and amylopectin, in which the a1±4 linked chains are highly branched by a1±6 linkages, which are formed by starch branching enzymes. Several isoforms of starch synthase are involved in starch formation and these can be classified according to their localization within the plastid; one isoform, granule-bound starch synthase I (GBSSI), is bound exclusively to the starch granule and others are localized predominantly in the soluble phase (stroma) of the plastid. In general, GBSSI is a protein of < 60 kDa. Through mutant analysis GBSSI is known to be solely responsible for the synthesis of amylose, because the waxy mutants of cereals, the amf mutant of potato, the lam mutants of pea and GBSSI antisense potato lines all involve the reduction or elimination of GBSSI activity and a specific reduction or elimination of amylose in the starch of the storage organs of these species [1±4].Among the starch synthase isoforms found in the plastid stroma of higher plants, starch synthase II (SSII) is also found bound to starch granules in significant amounts [5±9]. However, SSII cannot compensate for the effects of loss of GBSSI on amylose synthesis in amf mutant potatoes, indicating that SSII is not involved in amylose synthesis in vivo [8]. In the storage organs of different higher plants, SSII may make quite different quantitative contributions to total starch synthase activity. Mutant analysis in pea...

To provide information on the roles of the different forms of ADP-glucose pyrophosphorylase (AGPase) in barley (Hordeum vulgare) endosperm and the nature of the genes encoding their subunits, a mutant of barley, Risø 16, lacking cytosolic AGPase activity in the endosperm was identified. The mutation specifically abolishes the small subunit of the cytosolic AGPase and is attributable to a large deletion within the coding region of a previously characterized small subunit gene that we have called Hv.AGP.S.1. The plastidial AGPase activity in the mutant is unaffected. This shows that the cytosolic and plastidial small subunits of AGPase are encoded by separate genes. We purified the plastidial AGPase protein and, using amino acid sequence information, we identified the novel small subunit gene that encodes this protein. Studies of the Risø 16 mutant revealed the following. First, the reduced starch content of the mutant showed that a cytosolic AGPase is required to achieve the normal rate of starch synthesis. Second, the mutant makes both A-and B-type starch granules, showing that the cytosolic AGPase is not necessary for the synthesis of these two granule types. Third, analysis of the phylogenetic relationships between the various small subunit proteins both within and between species, suggest that the cytosolic AGPase single small subunit gene probably evolved from a leaf single small subunit gene.In the endosperm of all of the species of grasses so far investigated, there are both cytosolic and plastidial forms of the enzyme of ADP-Glc pyrophosphorylase (AGPase). However, there are indications that there may be differences between species in the relative amounts of the plastidial and cytosolic activities of AGPase in the endosperm and in the nature of the genes encoding their subunits. Studies of mutants of maize (Zea mays) show that the cytosolic form accounts for Ͼ95% of the total activity of AGPase in the endosperm (Denyer et al., 1996) and that this form is essential for normal rates of starch synthesis (Tsai and Nelson, 1966;Dickinson and Preiss, 1969). These studies also show that the plastidial and cytosolic forms of AGPase are encoded by different pairs of large and small subunit genes. For example, mutations in maize at the Bt2 locus specifically affect the cytosolic single small subunit (SSU) of AGPase and eliminate the activity of AGPase in the cytosol, but they do not affect the plastidial AGPase SSU or activity Hannah et al., 2001). For barley (Hordeum vulgare), the situation is less clear, but there is evidence to suggest that barley differs from maize in the following respects.First, the plastidial activity as a proportion of the total AGPase activity is considerably higher in barley endosperm (15%; Thorbjørnsen et al., 1996b) than in maize (Ͻ5%; Denyer et al., 1996). In maize endosperm, we calculate from published measurements of the rate of starch synthesis and total AGPase activity in maize (Singletary et al., 1997) that the plastidial AGPase activity alone is not sufficient to account for t...