Hydrology influences carbon flux through metabolic pathways in the hypolimnion of a Mediterranean reservoir

Global change is modifying meteorological and hydrological factors that influence the thermal regime of water bodies. These modifications can lead to longer stratification periods with enlarged hypolimnetic anoxic periods, which can promote heterotrophic anaerobic processes and alter reservoir carbon cycling. Here, we quantified aerobic and anaerobic heterotrophic processes (aerobic respiration, denitrification, iron and manganese reduction, sulfate reduction, and methanogenesis) on dissolved inorganic carbon (DIC) production in the hypolimnion of a Mediterranean reservoir (El Gergal, Spain) under two contrasting hydrological conditions: a wet year with heavy direct rainfall and frequent water inputs from upstream reservoirs, and a dry year with scarce rainfall and negligible water inputs. During the wet year, water inputs and rainfall induced low water column thermal stability and earlier turnover. By contrast, thermal stratification was longer and more stable during the dry year. During wet conditions, we observed lower DIC accumulation in the hypolimnion, mainly due to weaker sulfate reduction and methanogenesis. By contrast, longer stratification during the dry year promoted higher hypolimnetic DIC accumulation, resulting from enhanced methanogenesis and sulfate reduction, thus increasing methane emissions and impairing reservoir water quality. Aerobic respiration, denitrification and metal reduction produced a similar amount of DIC in the hypolimnion during the two studied years. All in all, biological and geochemical (calcite dissolution) processes explained most of hypolimnetic DIC accumulation during stratification regardless of the hydrological conditions, but there is still ~ 30% of hypolimnetic DIC production that cannot be explained by the processes contemplated in this study and the assumptions made.This research was funded by project Alter-C (PID2020-114024GB-C31, PID2020-114024GB-C32, PID2020-114024GB-C33) of Spanish Ministry of Science and Innovation (Spanish Research Agency, AEI). JJM-P was supported by a Spanish FPI grant (RE2018-083596). EMASESA staff provide essential technical support during field surveys. R.M. acknowledges funding from Generalitat de Catalunya through the Consolidated Research Group 2017SGR1124 and the CERCA programme. Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Funding for open access charge: Universidad de Málaga / CBUA

Enhanced error estimator based on a nearly equilibrated moving least squares recovery technique for FEM and XFEM

In this paper a new technique aimed to obtain accurate estimates of the error in energy norm using a moving least squares (MLS) recovery-based procedure is presented. We explore the capabilities of a recovery technique based on an enhanced MLS fitting, which directly provides continuous interpolated fields, to obtain estimates of the error in energy norm as an alternative to the superconvergent patch recovery (SPR). Boundary equilibrium is enforced using a nearest point approach that modifies the MLS functional. Lagrange multipliers are used to impose a nearly exact satisfaction of the internal equilibrium equation. The numerical results show the high accuracy of the proposed error estimator

Effects of tidal-forcing variations on tidal properties along a narrow convergent estuary

A 1D analytical framework is implemented in a narrow convergent estuary that is 78 km in length (the Guadiana, Southern Iberia) to evaluate the tidal dynamics along the channel, including the effects of neap-spring amplitude variations at the mouth. The close match between the observations (damping from the mouth to ∼ 30 km, shoaling upstream) and outputs from semi-closed channel solutions indicates that the M2 tide is reflected at the estuary head. The model is used to determine the contribution of reflection to the dynamics of the propagating wave. This contribution is mainly confined to the upper one third of the estuary. The relatively constant mean wave height along the channel (< 10% variations) partly results from reflection effects that also modify significantly the wave celerity and the phase difference between tidal velocity and elevation (contradicting the definition of an “ideal” estuary). Furthermore, from the mouth to ∼ 50 km, the variable friction experienced by the incident wave at neap and spring tides produces wave shoaling and damping, respectively. As a result, the wave celerity is largest at neap tide along this lower reach, although the mean water level is highest in spring. Overall, the presented analytical framework is useful for describing the main tidal properties along estuaries considering various forcings (amplitude, period) at the estuary mouth and the proposed method could be applicable to other estuaries with small tidal amplitude to depth ratio and negligible river discharge.info:eu-repo/semantics/publishedVersio

Guillain-Barré syndrome: a century of progress

In 1916, Guillain, Barré and Strohl reported on two cases of acute flaccid paralysis with high cerebrospinal fluid protein levels and normal cell counts — novel findings that identified the disease we now know as Guillain–Barré syndrome (GBS). 100 years on, we have made great progress with the clinical and pathological characterization of GBS. Early clinicopathological and animal studies indicated that GBS was an immune-mediated demyelinating disorder, and that severe GBS could result in secondary axonal injury; the current treatments of plasma exchange and intravenous immunoglobulin, which were developed in the 1980s, are based on this premise. Subsequent work has, however, shown that primary axonal injury can be the underlying disease. The association of Campylobacter jejuni strains has led to confirmation that anti-ganglioside antibodies are pathogenic and that axonal GBS involves an antibody and complement-mediated disruption of nodes of Ranvier, neuromuscular junctions and other neuronal and glial membranes. Now, ongoing clinical trials of the complement inhibitor eculizumab are the first targeted immunotherapy in GBS

Efficient recovery-based error estimation for the smoothed finite element method for smooth and singular linear elasticity

[EN] An error control technique aimed to assess the quality of smoothed finite element approximations is presented in this paper. Finite element techniques based on strain smoothing appeared in 2007 were shown to provide significant advantages compared to conventional finite element approximations. In particular, a widely cited strength of such methods is improved accuracy for the same computational cost. Yet, few attempts have been made to directly assess the quality of the results obtained during the simulation by evaluating an estimate of the discretization error. Here we propose a recovery type error estimator based on an enhanced recovery technique. The salient features of the recovery are: enforcement of local equilibrium and, for singular problems a ¿smooth + singular¿ decomposition of the recovered stress. We evaluate the proposed estimator on a number of test cases from linear elastic structural mechanics and obtain efficient error estimations whose effectivities, both at local and global levels, are improved compared to recovery procedures not implementing these features.Stephane Bordas would like to thank the partial financial support of the Royal Academy of Engineering and of the Leverhulme Trust for his Senior Research Fellowship Towards the next generation surgical simulators as well as the financial support for Octavio A. Gonzalez-Estrada and Stephane Bordas from the UK Engineering Physical Science Research Council (EPSRC) under grant EP/G042705/1 Increased Reliability for Industrially Relevant Automatic Crack Growth Simulation with the eXtended Finite Element Method. Stephane Bordas also thanks partial financial support of the European Research Council Starting Independent Research Grant (ERC Stg grant agreement No. 279578) and the FP7 Initial Training Network Funding under grant number 289361 "Integrating Numerical Simulation and Geometric Design Technology, INSIST". This work has been carried out within the framework of the research project DPI2010-20542 of the Ministerio de Ciencia e Innovacion (Spain). The financial support from Universitat Politecnica de Valencia, PROMETEO/2012/023 and Generalitat Valenciana are also acknowledged.González Estrada, OA.; Natarajan, S.; J.J. Ródenas; Nguyen-Xuan, H.; Bordas, S. (2013). Efficient recovery-based error estimation for the smoothed finite element method for smooth and singular linear elasticity. Computational Mechanics. 52(1):37-52. https://doi.org/10.1007/s00466-012-0795-6S3752521Liu GR, Dai KY, Nguyen TT (2006) A smoothed finite element method for mechanics problems. Comput Mech 39(6): 859–877. doi: 10.1007/s00466-006-0075-4Liu GR, Nguyen TT, Dai KY, Lam KY (2007) Theoretical aspects of the smoothed finite element method (SFEM). Int J Numer Methods Eng 71(8): 902–930Nguyen-Xuan H, Bordas SPA, Nguyen-Dang H (2008) Smooth finite element methods: convergence, accuracy and properties. Int J Numer Methods Eng 74(2): 175–208. doi: 10.1002/nmeBordas SPA, Natarajan S (2010) On the approximation in the smoothed finite element method (SFEM). Int J Numer Methods Eng 81(5): 660–670. doi: 10.1002/nmeZhang HH, Liu SJ, Li LX (2008) On the smoothed finite element method. Int J Numer Methods Eng 76(8): 1285–1295. doi: 10.1002/nme.2460Nguyen-Thoi T, Liu G, Lam K, Zhang G. (2009) A face-based smoothed finite element method (FS-FEM) for 3D linear and nonlinear solid mechanics using 4-node tetrahedral elements. Int J Numer Methods Eng 78: 324–353Liu G, Nguyen-Thoi T, Lam K (2009) An edge-based smoothed finite element method (ES-FEM) for static, free and forced vibration analyses of solids. J Sound Vib 320: 1100–1130Liu G, Nguyen-Thoi T, Nguyen-Xuan H, Lam K (2009) A node based smoothed finite element method (NS-FEM) for upper bound solution to solid mechanics problems. Comput Struct 87: 14–26Liu G. Smoothed Finite Element Methods. CRC Press, 2010Liu G, Nguyen-Xuan H, Nguyen-Thoi T (2010) A theoretical study on the smoothed FEM (SFEM) models: Properties, accuracy and convergence rates. Int J Numer Methods Biomed Eng 84: 1222–1256Nguyen T, Liu G, Dai K, Lam K (2007) smoothed finite element method. Tsinghua Sci Technol 12: 497–508Hung NX, Bordas S, Hung N (2009) Addressing volumetric locking and instabilities by selective integration in smoothed finite element. Commun Numer Methods Eng 25: 19–34Nguyen-Xuan H, Rabczuk T, Bordas S, Debongnie JF (2008) A smoothed finite element method for plate analysis. Comput Methods Appl Mech Eng 197: 1184–1203Nguyen NT, Rabczuk T, Nguyen-Xuan H, Bordas S (2008) A smoothed finite element method for shell analysis. Comput Methods Appl Mech Eng 198: 165–177Bordas SPA, Rabczuk T, Hung NX, Nguyen VP, Natarajan S, Bog T, óuan DM, Hiep NV (2010) Strain smoothing in FEM and XFEM. Comput Struct 88(23–24): 1419–1443. doi: 10.1016/j.compstruc.2008.07.006Bordas SP, Natarajan S, Kerfriden P, Augarde CE, Mahapatra DR, Rabczuk T, Pont SD (2011) On the performance of strain smoothing for óuadratic and enriched finite element approximations (XFEM/GFEM/PUFEM). Int J Numer Methods Biomed Eng 86: 637–666Liu G, Nguyen-Thoi T, Nguyen-Xuan H, Dai K, Lam K (2009) On the essence and the evaluation of the shape functions for the smoothed finite element method (SFEM). Int J Numer Methods Eng 77: 1863–1869. doi: 10.1002/nme.2587Strouboulis T, Zhang L, Wang D, Babuška I. (2006) A posteriori error estimation for generalized finite element methods. Comput Methods Appl Mech Eng 195(9–12): 852–879Bordas SPA, Duflot M (2007) Derivative recovery and a posteriori error estimate for extended finite elements. Comput Methods Appl Mech Eng 196(35–36): 3381–3399Xiao óZ, Karihaloo BL (2004) Statically admissible stress recovery using the moving least sóuares technique. In: Topping BHV, Soares CAM (eds) Progress in computational structures technology. Saxe-Coburg Publications, Stirling, pp 111–138Ródenas JJ, González-Estrada OA, Tarancón JE, Fuenmayor FJ (2008) A recovery-type error estimator for the extended finite element method based on singular + smooth stress field splitting. Int J Numer Methods Eng 76(4): 545–571. doi: 10.1002/nme.2313Panetier J, Ladevèze P, Chamoin L (2010) Strict and effective bounds in goal-oriented error estimation applied to fracture mechanics problems solved with XFEM. Int J Numer Methods Eng 81(6): 671–700Barros FB, Proenca SPB, de Barcellos CS (2004) On error estimator and p-adaptivity in the generalized finite element method. Int J Numer Methods Eng 60(14):2373–2398. doi: 10.1002/nme.1048Nguyen-Thoi T, Liu G, Nguyen-Xuan H, Nguyen-Tran C (2011) Adaptive analysis using the node-based smoothed finite element method (NS-FEM). Int J Numer Methods Biomed Eng 27(2): 198–218. doi: 10.1002/cnmGonzález-Estrada OA, Ródenas JJ, Bordas SPA, Duflot M, Kerfriden P, Giner E (2012) On the role of enrichment and statical admissibility of recovered fields in a-posteriori error estimation for enriched finite element methods. Eng Comput 29(8)Zienkiewicz OC, Zhu JZ (1987) A simple error estimator and adaptive procedure for practical engineering analysis. Int J Numer Methods Eng 24(2): 337–357Ródenas JJ, González-Estrada OA, Díez P, Fuenmayor FJ (2010) Accurate recovery-based upper error bounds for the extended finite element framework. Comput Methods Appl Mech Eng 199(37–40): 2607–2621Williams ML (1952) Stress singularities resulting from various boundary conditions in angular corners of plate in extension. J Appl Mech 19: 526–534Szabó BA, Babuška I (1991) Finite element analysis. Wiley, New YorkBarber JR. (2010) Elasticity. Series: solid mechanics and its application, 3rd edn. Springer, DordrechtChen JS, Wu CT, Yoon S, You Y (2001) A stabilized conforming nodal integration for Galerki mesh-free methods. Int J Numer Methods Eng 50: 435–466Yoo J, Moran B, Chen J (2004) Stabilized conforming nodal integration in the natural element method. Int J Numer Methods Eng 60: 861–890Zienkiewicz OC, Zhu JZ (1992) The superconvergent patch recovery and a posteriori error estimates. Part 1: The recovery technique. Int J Numer Methods Eng 33(7): 1331–1364Zienkiewicz OC, Zhu JZ (1992) The superconvergent patch recovery and a posteriori error estimates. Part 2: Error estimates and adaptivity. Int J Numer Methods Eng 33(7): 1365–1382Wiberg NE, Abdulwahab F (1993) Patch recovery based on superconvergent derivatives and eóuilibrium. Int J Numer Methods Eng 36(16): 2703–2724. doi: 10.1002/nme.1620361603Blacker T, Belytschko T (1994) Superconvergent patch recovery with eóuilibrium and conjoint interpolant enhancements. Int J Numer Methods Eng 37(3): 517–536Stein E, Ramm E, Rannacher R (2003) Error-controlled adaptive finite elements in solid mechanics. Wiley, ChichesterDuflot M, Bordas SPA (2008) A posteriori error estimation for extended finite elements by an extended global recovery. 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In: Khalili N, Valliappan S, Li ó, Russell A (eds) 9th World congress on computational mechanics (WCCM9). 4th Asian Pacific Congress on computational methods (APCOM2010). Centre for Infrastructure Engineering and SafetyRódenas JJ, González-Estrada OA, Díez P, Fuenmayor FJ (2007) Upper bounds of the error in the extended finite element method by using an eóuilibrated-stress patch recovery technique. In: International conference on adaptive modeling and simulation (ADMOS 2007). International Center for Numerical Methods in Engineering (CIMNE), pp 210–213Menk A, Bordas S (2010) Numerically determined enrichment function for the extended finite element method and applications to bi-material anisotropic fracture and polycrystals. Int J Numer Methods Eng 83: 805–828Menk A, Bordas S (2011) Crack growth calculations in solder joints based on microstructural phenomena with X-FEM. 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Mammalian Stem Cells Reprogramming in Response to Terahertz Radiation

We report that extended exposure to broad-spectrum terahertz radiation results in specific changes in cellular functions that are closely related to DNA-directed gene transcription. Our gene chip survey of gene expression shows that whereas 89% of the protein coding genes in mouse stem cells do not respond to the applied terahertz radiation, certain genes are activated, while other are repressed. RT-PCR experiments with selected gene probes corresponding to transcripts in the three groups of genes detail the gene specific effect. The response was not only gene specific but also irradiation conditions dependent. Our findings suggest that the applied terahertz irradiation accelerates cell differentiation toward adipose phenotype by activating the transcription factor peroxisome proliferator-activated receptor gamma (PPARG). Finally, our molecular dynamics computer simulations indicate that the local breathing dynamics of the PPARG promoter DNA coincides with the gene specific response to the THz radiation. We propose that THz radiation is a potential tool for cellular reprogramming

Benefits of a ball and chain: simple environmental enrichments improve welfare and reproductive success in farmed American mink (Neovison vison)

Can simple enrichments enhance caged mink welfare? Pilot data from 756 sub-adults spanning three colour-types (strains) identified potentially practical enrichments, and suggested beneficial effects on temperament and fur-chewing. Our main experiment started with 2032 Black mink on three farms: from each of 508 families, one juvenile male-female pair was enriched (E) with two balls and a hanging plastic chain or length of hose, while a second pair was left as a non-enriched (NE) control. At 8 months, more than half the subjects were killed for pelts, and 302 new females were recruited (half enriched: ‘late E’). Several signs of improved welfare or productivity emerged. Access to enrichment increased play in juveniles. E mink were calmer (less aggressive in temperament tests; quieter when handled; less fearful, if male), and less likely to fur-chew, although other stereotypic behaviours were not reduced. On one farm, E females had lower cortisol (inferred from faecal metabolites). E males tended to copulate for longer. E females also weaned more offspring: about 10% more juveniles per E female, primarily caused by reduced rates of barrenness (‘late E’ females also giving birth to bigger litters on one farm), effects that our data cautiously suggest were partly mediated by reduced inactivity and changes in temperament. Pelt quality seemed unaffected, but E animals had cleaner cages. In a subsidiary side-study using 368 mink of a second colour-type (‘Demis’), similar temperament effects emerged, and while E did not reduce fur-chewing or improve reproductive success in this colour-type, E animals were judged to have better pelts. Overall, simple enrichments were thus beneficial. These findings should encourage welfare improvements on fur farms (which house 60-70 million mink p.a.) and in breeding centres where endangered mustelids (e.g. black-footed ferrets) often reproduce poorly. They should also stimulate future research into more effective practical enrichments

Environmental enrichment reduces signs of boredom in caged mink

Animals housed in impoverished cages are often labelled 'bored'. They have also been called 'apathetic' or 'depressed', particularly when profoundly inactive. However, these terms are rarely operationally defined and validated. As a negative state caused by under-stimulation, boredom should increase interest in stimuli of all kinds. Apathy (lack of interest), by contrast, should manifest as decreased interest in all stimuli, while anhedonia (loss of pleasure, a depressive symptom) should specifically decrease interest in normally rewarding stimuli. We tested the hypotheses that mink, a model carnivore, experience more boredom, depression-like apathy, or anhedonia in non-enriched (NE) cages than in complex, enriched (E) cages. We exposed 29 subjects (13 E, 16 NE) to ten stimuli categorized a priori as aversive (e.g. air puffs), rewarding (e.g. evoking chasing) or ambiguous/neutral (e.g. candles). Interest in stimuli was assessed via latencies to contact, contact durations, and durations oriented to stimuli. NE mink contacted all stimuli faster (P = 0.003) than E mink, and spent longer oriented to/in contact with them, albeit only significantly so for ambiguous ones (treatment*type P<0.013). With stimulus category removed from statistical models, interest in all stimuli was consistently higher among NE mink (P<0.0001 for all measures). NE mink also consumed more food rewards (P = 0.037). Finally, we investigated whether lying down while awake and stereotypic behaviour (both increased by NE housing) predicted these responses. Lying awake positively co-varied with certain measures of increased exploration. In contrast, stereotypic 'scrabbling' or locomotion (e.g. pacing) did not. Overall, NE mink showed no evidence of apathy or depression, but instead a heightened investigation of diverse stimuli consistent with boredom. This state was potentially indicated by spending much time lying still but awake (although this result requires replication). Boredom can thus be operationalized and assessed empirically in non-human animals. It can also be reduced by environmental enrichment

Natural History of MYH7-Related Dilated Cardiomyopathy

BACKGROUND: Variants in myosin heavy chain 7 (MYH7) are responsible for disease in 1% to 5% of patients with dilated cardiomyopathy (DCM); however, the clinical characteristics and natural history of MYH7-related DCM are poorly described. OBJECTIVE: We sought to determine the phenotype and prognosis of MYH7-related DCM. We also evaluated the influence of variant location on phenotypic expression. METHODS: We studied clinical data from 147 individuals with DCM-causing MYH7 variants (47.6% female; 35.6 ± 19.2 years) recruited from 29 international centers. RESULTS: At initial evaluation, 106 (72.1%) patients had DCM (left ventricular ejection fraction: 34.5% ± 11.7%). Median follow-up was 4.5 years (IQR: 1.7-8.0 years), and 23.7% of carriers who were initially phenotype-negative developed DCM. Phenotypic expression by 40 and 60 years was 46% and 88%, respectively, with 18 patients (16%) first diagnosed at <18 years of age. Thirty-six percent of patients with DCM met imaging criteria for LV noncompaction. During follow-up, 28% showed left ventricular reverse remodeling. Incidence of adverse cardiac events among patients with DCM at 5 years was 11.6%, with 5 (4.6%) deaths caused by end-stage heart failure (ESHF) and 5 patients (4.6%) requiring heart transplantation. The major ventricular arrhythmia rate was low (1.0% and 2.1% at 5 years in patients with DCM and in those with LVEF of ≤35%, respectively). ESHF and major ventricular arrhythmia were significantly lower compared with LMNA-related DCM and similar to DCM caused by TTN truncating variants. CONCLUSIONS: MYH7-related DCM is characterized by early age of onset, high phenotypic expression, low left ventricular reverse remodeling, and frequent progression to ESHF. Heart failure complications predominate over ventricular arrhythmias, which are rare

SILEX: a fast and inexpensive high-quality DNA extraction method suitable for multiple sequencing platforms and recalcitrant plant species

[EN] Background The use of sequencing and genotyping platforms has undergone dramatic improvements, enabling the generation of a wealth of genomic information. Despite this progress, the availability of high-quality genomic DNA (gDNA) in sufficient concentrations is often a main limitation, especially for third-generation sequencing platforms. A variety of DNA extraction methods and commercial kits are available. However, many of these are costly and frequently give either low yield or low-quality DNA, inappropriate for next generation sequencing (NGS) platforms. Here, we describe a fast and inexpensive DNA extraction method (SILEX) applicable to a wide range of plant species and tissues. Results SILEX is a high-throughput DNA extraction protocol, based on the standard CTAB method with a DNA silica matrix recovery, which allows obtaining NGS-quality high molecular weight genomic plant DNA free of inhibitory compounds. SILEX was compared with a standard CTAB extraction protocol and a common commercial extraction kit in a variety of species, including recalcitrant ones, from different families. In comparison with the other methods, SILEX yielded DNA in higher concentrations and of higher quality. Manual extraction of 48 samples can be done in 96 min by one person at a cost of 0.12 euro/sample of reagents and consumables. Hundreds of tomato gDNA samples obtained with either SILEX or the commercial kit were successfully genotyped with Single Primer Enrichment Technology (SPET) with the Illumina HiSeq 2500 platform. Furthermore, DNA extracted fromSolanum elaeagnifoliumusing this protocol was assessed by Pulsed-field gel electrophoresis (PFGE), obtaining a suitable size ranges for most sequencing platforms that required high-molecular-weight DNA such as Nanopore or PacBio. Conclusions A high-throughput, fast and inexpensive DNA extraction protocol was developed and validated for a wide variety of plants and tissues. SILEX offers an easy, scalable, efficient and inexpensive way to extract DNA for various next-generation sequencing applications including SPET and Nanopore among others.This research has been funded by the European Union's Horizon 2020 research and innovation programme under grant agreement No 677379 (Linking genetic resources, genomes and phenotypes of Solanaceous crops; G2P-SOL). David Alonso is grateful to Universitat Politecnica de Valencia for a predoctoral (PAID-01-16) contract under the Programa de Ayudas de Investigacion y Desarrollo initiative. Mariola Plazas is grateful to Generalitat Valenciana and Fondo Social Europeo for a postdoctoral grant (APOSTD/2018/014). Pietro Gramazio is grateful to Japan Society for the Promotion of Science for a Postdoctoral Grant (P19105, FY2019 JSPS Postdoctoral Fellowship for Research in Japan (Standard)). The Spanish Ministerio de Educacion, Cultura y Deporte funded a predoctoral fellowship granted to Edgar Garcia-Fortea (FPU17/02389).Vilanova Navarro, S.; Alonso-Martín, D.; Gramazio, P.; Plazas Ávila, MDLO.; García-Fortea, E.; Ferrante, P.; Schmidt, M.... (2020). SILEX: a fast and inexpensive high-quality DNA extraction method suitable for multiple sequencing platforms and recalcitrant plant species. Plant Methods. 16(1):1-11. https://doi.org/10.1186/s13007-020-00652-yS111161Scheben A, Batley J, Edwards D. Genotyping-by-sequencing approaches to characterize crop genomes: choosing the right tool for the right application. Plant Biotechnol J. 2017;15:149–61.Jung H, Winefield C, Bombarely A, Prentis P, Waterhouse P. Tools and strategies for long-read sequencing and de novo assembly of plant genomes. 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