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Highly parallel SNP genotyping platforms have been developed for some important crop species, but these platforms typically carry a high cost per sample for first-time or small-scale users. In contrast, recently developed genotyping by sequencing (GBS) approaches offer a highly cost effective alternative for simultaneous SNP discovery and genotyping. In the present investigation, we have explored the use of GBS in soybean. In addition to developing a novel analysis pipeline to call SNPs and indels from the resulting sequence reads, we have devised a modified library preparation protocol to alter the degree of complexity reduction. We used a set of eight diverse soybean genotypes to conduct a pilot scale test of the protocol and pipeline. Using ApeKI for GBS library preparation and sequencing on an Illumina GAIIx machine, we obtained 5.5 M reads and these were processed using our pipeline. A total of 10,120 high quality SNPs were obtained and the distribution of these SNPs mirrored closely the distribution of gene-rich regions in the soybean genome. A total of 39.5% of the SNPs were present in genic regions and 52.5% of these were located in the coding sequence. Validation of over 400 genotypes at a set of randomly selected SNPs using Sanger sequencing showed a 98% success rate. We then explored the use of selective primers to achieve a greater complexity reduction during GBS library preparation. The number of SNP calls could be increased by almost 40% and their depth of coverage was more than doubled, thus opening the door to an increase in the throughput and a significant decrease in the per sample cost. The approach to obtain high quality SNPs developed here will be helpful for marker assisted genomics as well as assessment of available genetic resources for effective utilisation in a wide number of species.

From 1834 to 1850, Latin America's first penitentiary, the Casa de Correção in Rio de Janeiro, was a construction site where slaves, “liberated Africans”, convicts, and unfree workers interacted daily, forged identities, and deployed resistance strategies against the pressures of confinement and the demands of Brazil's eclectic labor regimes. This article examines the utilization of this motley crew of workers, the interactions among “liberated Africans”, slaves, and convict laborers, and the government's intervention between 1848 and 1850 to restrict slave labor at the prison in favor of free waged workers. It asserts that the abolition of the slave trade in 1850 and the subsequent inauguration of the penitentiary augured profound changes in Rio's labor landscape, from a predominantly unfree to a free wage labor force.

Estimating sugarcane biomass is difficult to achieve when working with highly variable spatial distributions of growing conditions, like on Reunion Island. We used a dataset of in-farm fields with contrasted climatic conditions and farming practices to compare three methods of yield estimation based on remote sensing: (1) an empirical relationship method with a growing season-integrated Normalized Difference Vegetation Index NDVI, (2) the Kumar-Monteith efficiency model, and (3) a forced-coupling method with a sugarcane crop model (MOSICAS) and satellite-derived fraction of absorbed photosynthetically active radiation. These models were compared with the crop model alone and discussed to provide recommendations for a satellite-based system for the estimation of yield at the field scale. Results showed that the linear empirical model produced the best results (RMSE = 10.4 t∙ha−1). Because this method is also the simplest to set up and requires less input data, it appears that it is the most suitable for performing operational estimations and forecasts of sugarcane yield at the field scale. The main limitation is the acquisition of a minimum of five satellite images. The upcoming open-access Sentinel-2 Earth observation system should overcome this limitation because it will provide 10-m resolution satellite images with a 5-day frequency.

Grain size is a key agronomic trait that contributes to grain yield in hexaploid wheat. Grain length and width were evaluated in an international collection of 157 wheat accessions. These accessions were genetically characterized using a genotyping-by-sequencing (GBS) protocol that produced 73,784 single nucleotide polymorphism (SNP) markers. GBS-derived genotype calls obtained on Chinese Spring proved extremely accurate when compared to the reference (> 99.9%) and showed > 95% agreement with calls made at SNP loci shared with the 90 K SNP array on a subset of 71 Canadian wheat accessions for which both types of data were available. This indicates that GBS can yield a large amount of highly accurate SNP data in hexaploid wheat. The genetic diversity analysis performed using this set of SNP markers revealed the presence of six distinct groups within this collection. A GWAS was conducted to uncover genomic regions controlling variation for grain length and width. In total, seven SNPs were found to be associated with one or both traits, identifying three quantitative trait loci (QTLs) located on chromosomes 1D, 2D and 4A. In the vicinity of the peak SNP on chromosome 2D, we found a promising candidate gene (TraesCS2D01G331100), whose rice ortholog (D11) had previously been reported to be involved in the regulation of grain size. These markers will be useful in breeding for enhanced wheat productivity.

Genotyping-by-sequencing (GBS) is a relatively low-cost high throughput genotyping technology based on next generation sequencing and is applicable to orphan species with no reference genome. A combination of genome complexity reduction and multiplexing with DNA barcoding provides a simple and affordable way to resolve allelic variation between plant samples or populations. GBS was performed on ApeKI libraries using DNA from 48 genotypes each of two heterogeneous populations of tetraploid alfalfa (Medicago sativa spp. sativa): the synthetic cultivar Apica (ATF0) and a derived population (ATF5) obtained after five cycles of recurrent selection for superior tolerance to freezing (TF). Nearly 400 million reads were obtained from two lanes of an Illumina HiSeq 2000 sequencer and analyzed with the Universal Network-Enabled Analysis Kit (UNEAK) pipeline designed for species with no reference genome. Following the application of whole dataset-level filters, 11,694 single nucleotide polymorphism (SNP) loci were obtained. About 60% had a significant match on the Medicago truncatula syntenic genome. The accuracy of allelic ratios and genotype calls based on GBS data was directly assessed using 454 sequencing on a subset of SNP loci scored in eight plant samples. Sequencing depth in this study was not sufficient for accurate tetraploid allelic dosage, but reliable genotype calls based on diploid allelic dosage were obtained when using additional quality filtering. Principal Component Analysis of SNP loci in plant samples revealed that a small proportion (<5%) of the genetic variability assessed by GBS is able to differentiate ATF0 and ATF5. Our results confirm that analysis of GBS data using UNEAK is a reliable approach for genome-wide discovery of SNP loci in outcrossed polyploids.

[...]more durable sources of resistance are needed to manage P. sojae. [...]RNA‐seq, as a genotyping tool, represents an alternative reduced‐representation approach focusing on protein‐coding regions (Scheben et al., ). [...]this study proposes an integrated approach, exploiting a new phenotyping procedure, RNA‐seq analyses and SNP variants of predicted functional impact, to discriminate and prioritize high‐value candidate genes modulating complex quantitative traits such as those defining PR during plant–pathogen interactions.

The SoyaGen project was a collaborative endeavor involving Canadian soybean researchers and breeders from academia and the private sector as well as international collaborators. Its aims were to develop genomics-derived solutions to real-world challenges faced by breeders. Based on the needs expressed by the stakeholders, the research efforts were focused on maximizing realized yield through optimization of maturity and improved disease resistance. The main deliverables related to molecular breeding in soybean will be reviewed here. These include: (1) SNP datasets capturing the genetic diversity within cultivated soybean (both within a worldwide collection of > 1,000 soybean accessions and a subset of 102 short-season accessions (MG0 and earlier) directly relevant to this group); (2) SNP markers for selecting favorable alleles at key maturity genes as well as loci associated with increased resistance to key pathogens and pests ( Phytophthora sojae , Heterodera glycines , Sclerotinia sclerotiorum ); (3) diagnostic tools to facilitate the identification and mapping of specific pathotypes of P. sojae ; and (4) a genomic prediction approach to identify the most promising combinations of parents. As a result of this fruitful collaboration, breeders have gained new tools and approaches to implement molecular, genomics-informed breeding strategies. We believe these tools and approaches are broadly applicable to soybean breeding efforts around the world.

Small-molecule inhibitor of apoptosis (IAP) antagonists, called Smac mimetic compounds (SMCs), sensitize tumours to TNF-α-induced killing while simultaneously blocking TNF-α growth-promoting activities. SMCs also regulate several immunomodulatory properties within immune cells. We report that SMCs synergize with innate immune stimulants and immune checkpoint inhibitor biologics to produce durable cures in mouse models of glioblastoma in which single agent therapy is ineffective. The complementation of activities between these classes of therapeutics is dependent on cytotoxic T-cell activity and is associated with a reduction in immunosuppressive T-cells. Notably, the synergistic effect is dependent on type I IFN and TNF-α signalling. Furthermore, our results implicate an important role for TNF-α-producing cytotoxic T-cells in mediating the anti-cancer effects of immune checkpoint inhibitors when combined with SMCs. Overall, this combinatorial approach could be highly effective in clinical application as it allows for cooperative and complimentary mechanisms in the immune cell-mediated death of cancer cells. Smac mimetics sensitize cancer cells to the extrinsic cell death pathway and stimulate anti-tumour immunity. In this study, the authors show that Smac mimetics can synergize with immune checkpoint inhibitors to control tumour growth in mouse cancer models, including aggressive CNS tumours, in a cytotoxic CD8 + T-cell- and TNFα-dependent manner.

Agro‐Land Surface Models (agro‐LSM) combine detailed crop models and large‐scale vegetation models (DGVMs) to model the spatial and temporal distribution of energy, water, and carbon fluxes within the soil–vegetation–atmosphere continuum worldwide. In this study, we identify and optimize parameters controlling leaf area index (LAI) in the agro‐LSM ORCHIDEE‐STICS developed for sugarcane. Using the Morris method to identify the key parameters impacting LAI, at eight different sugarcane field trial sites, in Australia and La Reunion island, we determined that the three most important parameters for simulating LAI are (i) the maximum predefined rate of LAI increase during the early crop development phase, a parameter that defines a plant density threshold below which individual plants do not compete for growing their LAI, and a parameter defining a threshold for nitrogen stress on LAI. A multisite calibration of these three parameters is performed using three different scoring functions. The impact of the choice of a particular scoring function on the optimized parameter values is investigated by testing scoring functions defined from the model‐data RMSE, the figure of merit and a Bayesian quadratic model‐data misfit function. The robustness of the calibration is evaluated for each of the three scoring functions with a systematic cross‐validation method to find the most satisfactory one. Our results show that the figure of merit scoring function is the most robust metric for establishing the best parameter values controlling the LAI. The multisite average figure of merit scoring function is improved from 67% of agreement to 79%. The residual error in LAI simulation after the calibration is discussed.

Estimation of allelic frequencies is often required in breeding but genotyping many individuals at many loci can be expensive. We have developed a genotyping‐by‐sequencing (GBS) approach for estimating allelic frequencies on pooled samples (Pool‐GBS) and used it to examine segregation distortion in doubled haploid (DH) populations of barley (Hordeum vulgare L.). In the first phase, we genotyped each line individually and exploited these data to explore a strategy to call single nucleotide polymorphisms (SNPs) on pooled reads. We measured both the number of SNPs called and the variance of the estimated allelic frequencies at various depths of coverage on a subset of reads containing 5 to 25 million reads. We show that allelic frequencies could be cost‐effectively and accurately estimated at a depth of 50 reads per SNP using 15 million reads. This Pool‐GBS approach yielded 1984 SNPs whose allelic frequency estimates were highly reproducible (CV = 10.4%) and correlated (r = 0.9167) with the “true” frequency derived from analysis of individual lines. In a second phase, we used Pool‐GBS to investigate segregation bias throughout androgenesis from microspores to a population of regenerated plants. No strong bias was detected among the microspores resulting from the meiotic divisions, whereas significant biases could be shown to arise during embryo formation and plant regeneration. In summary, this methodology provides an approach to estimate allelic frequencies more efficiently and on materials that are unsuitable for individual analysis. In addition, it allowed us to shed light on the process of androgenesis in barley.