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Unpredictable dynamics arising from a sensitivity to initial conditions is commonly associated with chaos. We demonstrate how similar unpredictability manifests in a nonlinear system that possesses a large number of long-term outcomes, namely the propagation of an air bubble within a viscous fluid-filled channel. The system under investigation supports various stable states of single-bubble propagation. In addition, bubbles can readily break up during their propagation. Upon subjecting steadily-propagating bubbles to finite-amplitude perturbations in the form of localised channel constrictions, we identify localised regions of the driving flow rate for which the resulting evolutions are unpredictable. Visibly-indistinguishable bubbles are observed to evolve towards a multitude of long-term outcomes, including each of the stable states available to the initial bubble and various states of permanently-changed bubble topology. By combining high-precision experimental results with simulations of a depth-averaged lubrication model of the system, we determine that this behaviour is driven by a sensitive dependence on initial conditions within the vicinity of an unstable periodic orbit.
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Introduction: Despite its rapid worldwide adoption as an efficient mutagenesis tool, plant genome editing remains a labor-intensive process requiring often several months of in vitro culture to obtain mutant plantlets. To avoid a waste in time and money and to test, in only a few days, the efficiency of molecular constructs or novel Cas9 variants (clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9) prior to stable transformation, rapid analysis tools are helpful. Methods: To this end, a streamlined maize protoplast system for transient expression of CRISPR/Cas9 tools coupled to NGS (next generation sequencing) analysis and a novel bioinformatics pipeline was established. Results and discussion: Mutation types found with high frequency in maize leaf protoplasts had a trend to be the ones observed after stable transformation of immature maize embryos. The protoplast system also allowed to conclude that modifications of the sgRNA (single guide RNA) scaffold leave little room for improvement, that relaxed PAM (protospacer adjacent motif) sites increase the choice of target sites for genome editing, albeit with decreased frequency, and that efficient base editing in maize could be achieved for certain but not all target sites. Phenotypic analysis of base edited mutant maize plants demonstrated that the introduction of a stop codon but not the mutation of a serine predicted to be phosphorylated in the bHLH (basic helix loop helix) transcription factor ZmICEa (INDUCER OF CBF EXPRESSIONa) caused abnormal stomata, pale leaves and eventual plant death two months after sowing.
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Isolated microspore culture systems have been designed in maize by several groups, mainly from the late 1980s to early 2000s. However, even with optimized protocols, microspore embryogenesis induction has remained very dependent on the genotype in maize, with elite germplasm generally displaying no response or very low response. Yet, these last few years, significant progress has been accomplished in understanding and controlling microspore embryogenesis induction in model dicot and monocot species. This knowledge may be transferred to maize, and isolated microspore culture may gain new interest in this crop, at least for embryogenesis research. The methods we hereby present in detail permit the purification of 3-12 × 105 viable microspores per maize tassel, at the favorable stage for microspore embryogenesis. When cultured in appropriate liquid media, microspores from responsive genotypes give rise to androgenic embryos, which can then be regenerated into fertile doubled haploid plants.
Assuntos
Gametogênese Vegetal , Técnicas de Cultura de Tecidos/métodos , Zea mays/crescimento & desenvolvimento , Meios de Cultura , Haploidia , Pólen/embriologia , Pólen/genética , Pólen/crescimento & desenvolvimento , Zea mays/embriologia , Zea mays/genéticaRESUMO
Identification of quantitative trait loci (QTL) involved in the variation of hybrid value is of key importance for cross-pollinated species such as maize (Zea mays L.). In a companion paper, we illustrated a new QTL mapping population design involving a factorial mating between two multiparental segregating populations. Six biparental line populations were developed from four founder lines in the Dent and Flint heterotic groups. They were crossed to produce 951 hybrids and evaluated for silage performances. Previously, a linkage analysis (LA) model that assumes each founder line carries a different allele was used to detect QTL involved in General and Specific Combining Abilities (GCA and SCA, respectively) of hybrid value. This previously introduced model requires the estimation of numerous effects per locus, potentially affecting QTL detection power. Using the same design, we compared this "Founder alleles" model to two more parsimonious models, which assume that (i) identity in state at SNP alleles from the same heterotic group implies identity by descent (IBD) at linked QTL ("SNP within-group" model) or (ii) identity in state implies IBD, regardless of population origin of the alleles ("Hybrid genotype" model). This last model assumes biallelic QTL with equal effects in each group. It detected more QTL on average than the two other models but explained lower percentages of variance. The "SNP within-group" model appeared to be a good compromise between the two other models. These results confirm the divergence between the Dent and Flint groups. They also illustrate the need to adapt the QTL detection model to the complexity of the allelic variation, which depends on the trait, the QTL, and the divergence between the heterotic groups.
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Biomassa , Hibridização Genética , Melhoramento Vegetal/métodos , Locos de Características Quantitativas , Zea mays/genética , Mapeamento Cromossômico/métodos , Ligação Genética , Polimorfismo de Nucleotídeo Único , Zea mays/crescimento & desenvolvimentoRESUMO
Several plant and animal species of agricultural importance are commercialized as hybrids to take advantage of the heterosis phenomenon. Understanding the genetic architecture of hybrid performances is therefore of key importance. We developed two multiparental maize (Zea mays L.) populations, each corresponding to an important heterotic group (dent or flint) and comprised of six connected biparental segregating populations of inbred lines (802 and 822 lines for each group, respectively) issued from four founder lines. Instead of using "testers" to evaluate their hybrid values, segregating lines were crossed according to an incomplete factorial design to produce 951 dent-flint hybrids, evaluated for four biomass production traits in eight environments. QTL detection was carried out for the general-combining-ability (GCA) and specific-combining-ability (SCA) components of hybrid value, considering allelic effects transmitted from each founder line. In total, 42 QTL were detected across traits. We detected mostly QTL affecting GCA, 31% (41% for dry matter yield) of which also had mild effects on SCA. The small impact of dominant effects is consistent with the known differentiation between the dent and flint heterotic groups and the small percentage of hybrid variance due to SCA observed in our design (â¼20% for the different traits). Furthermore, most (80%) of GCA QTL were segregating in only one of the two heterotic groups. Relative to tester-based designs, use of hybrids between two multiparental populations appears highly cost efficient to detect QTL in two heterotic groups simultaneously. This presents new prospects for selecting superior hybrid combinations with markers.
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Hibridização Genética , Modelos Genéticos , Locos de Características Quantitativas , Zea mays/genética , Biomassa , Genes Dominantes , Variação Genética , Endogamia , Característica Quantitativa Herdável , Zea mays/crescimento & desenvolvimentoRESUMO
When air is blown in a straw or tube near an air-liquid interface, typically one of two behaviors is observed: a dimple in the liquid's surface, or a frenzy of sputtering bubbles, waves, and spray. Here we report and characterize an intermediate regime that can develop when a confined air jet enters the interface at an angle. This regime is oscillatory with a distinct characteristic frequency and can develop periodic angled jets that can break up into monodisperse aerosols. The underlying mechanisms responsible for this highly periodic regime are not well understood. Here we flow a continuous stream of gas through a tube near a liquid surface, observing both optically and acoustically the deformation of the liquid-air interface as various parameters are systematically adjusted. We show that the Kelvin-Helmholtz instability is responsible for the inception of waves within a cavity formed by the gas. Inertia, gravity, and capillary forces both shape the cavity and govern the frequency and amplitude of these gas-induced cavity waves. The flapping cavity focuses the waves into a series of periodic jets that can break up into droplets following the Rayleigh-Plateau instability. We present scaling arguments to rationalize the fundamental frequencies driving this system, as well as the conditions that bound the periodic regime. These frequencies and conditions compare well with our experimental results.