Large Embryos Research Articles

Assessment of the Morphological Structure of Watermelon and Muskmelon Seeds as Related to Viability

Morphologically perfect seeds are critically important for raising normal, healthy seedlings as well as for increasing germination percentages. In this study, an X-ray microCT scanner was employed to generate CT images to investigate the internal conditions of naturally aged watermelon and muskmelon seeds. Different seed components, such as the seed coat thickness, the size of the embryo, the area of the air space inside the seed, the embryo structure, and the damages diagnosis in the seed anatomy were selected as observation parameters for investigation of the dicot seed samples. The areas of the seed coat thickness, embryo, and the internal air space were measured by ImageJ software, and the one-way analysis of variance (ANOVA) test was performed to inspect the significant differences in the parameters of the normal viable and the nonviable/abnormal viable seeds. A paper germination test was performed to identify the normal viable and nonviable/abnormal viable seeds. The findings of this study indicate that there was no significant difference in the seed coat thickness of the normal viable and the nonviable/abnormal viable naturally aged seeds, but the embryo and internal air space area structures were distinctive. The normal viable seeds contained a large embryo with nominal air space inside the seed, whereas the nonviable/abnormal viable seed embryo size was relatively small with a vast internal cavity. Based on the results of this study, we concluded that the normal viable and nonviable/abnormal viable aged seeds had significant differences in their morphological structures that adversely affect seed germination.

Twin formation from a twin boundary in Mg during in-situ nanomechanical testing

An important fundamental question regarding deformation twinning is whether it is possible for twins to nucleate at boundaries or interfaces when specific stress fields are present. A corollary that follows from this question is: if this is indeed possible, what determines the proper stress field and how does it occur at the nanoscale? Here, we demonstrate the application of an in-situ nanoindentation approach to confine and dynamically capture the stages in the formation of a deformation twin at an internal twin boundary in single crystal Mg. We observe the formation of contraction twin embryos at the pre-existing extension twin boundary, and the subsequent propagation of the twin embryos into the crystal. We reveal an intermediate step, involving the coalescence of tiny embryos into a larger embryo before the nucleus emanates into the crystal. De-twinnning of the twin embryos is captured during unloading and shown to leave a remnant nanosized twin (<10 nm) after complete unloading. A 3D full-field, crystal plasticity model identifies that the twin type and variant selection of the stable embryo are governed by the internal local stress state prevailing at the pre-existing twin boundary. The possible role played by <c+a> dislocations and boundary structure (incoherent vs. coherent) in embryo formation, as suggested by the TEM and modeling analyses, are discussed.

Identification and Characterization of LARGE EMBRYO, a New Gene Controlling Embryo Size in Rice (Oryza sativa L.)

BackgroundAlthough embryo accounts for only 2–3% of the total weight of a rice grain, it is a good source of various nutrients for human health. Because enlarged embryo size causes increase of the amount of nutrients and bioactive compounds stored within rice grain, giant embryo mutants of rice (Oryza sativa L.) are excellent genetic resources for improving the nutritional value of rice grains.ResultsThree giant embryo mutants, including large embryo (le), giant embryo (ge) and super-giant embryo (ges), with variable embryo size were used in this study. We investigated whether genes controlling embryo size in these mutants (le, ge and ges) were allelic to each other. Although ge and ges was allelic to GIANT EMBRY (GE), le was not allelic to ge and ges in allelism test. The GE gene carried a unique nucleotide substitution in each of the two mutants (ge and ges), resulting in non-synonymous mutations in exon 2 of GE in both mutants. However, the GE gene of the le mutant did not carry any mutation, suggesting that the enlarged embryo phenotype of le was governed by another gene. Using map-based cloning, we mapped the LE gene to the short arm of chromosome 3. The le mutant showed mild enlargement in embryo size, which resulted from an increase in the size of scutellar parenchyma cells. The LE encodes a C3HC4-type RING finger protein and was expressed to relatively high levels in seeds at a late developmental stage. Knockdown of LE expression using RNA interference increased the embryo size of rice grains, confirming the role of LE in determining the embryo size.ConclusionOverall, we identified a new gene controlling embryo size in rice. Phenotypic and molecular characterization results suggest that the le mutant will serve as a valuable resource for developing new rice cultivars with large embryos and nutrient-dense grains.

Growth after the streaming instability

Context. Streaming instability is a key mechanism in planet formation, clustering pebbles into planetesimals with the help of self-gravity. It is triggered at a particular disk location where the local volume density of solids exceeds that of the gas. After their formation, planetesimals can grow into protoplanets by feeding from other planetesimals in the birth ring as well as by accreting inwardly drifting pebbles from the outer disk. Aims. We aim to investigate the growth of planetesimals into protoplanets at a single location through streaming instability. For a solar-mass star, we test the conditions under which super-Earths are able to form within the lifetime of the gaseous disk. Methods. We modified the Mercury N-body code to trace the growth and dynamical evolution of a swarm of planetesimals at a distance of 2.7 AU from the star. The code simulates gravitational interactions and collisions among planetesimals, gas drag, type I torque, and pebble accretion. Three distributions of planetesimal sizes were investigated: (i) a mono-dispersed population of 400 km radius planetesimals, (ii) a poly-dispersed population of planetesimals from 200 km up to 1000 km, (iii) a bimodal distribution with a single runaway body and a swarm of smaller, 100 km size planetesimals. Results. The mono-dispersed population of 400 km size planetesimals cannot form protoplanets of a mass greater than that of the Earth. Their eccentricities and inclinations are quickly excited, which suppresses both planetesimal accretion and pebble accretion. Planets can form from the poly-dispersed and bimodal distributions. In these circumstances, it is the two-component nature that damps the random velocity of the large embryo through the dynamical friction of small planetesimals, allowing the embryo to accrete pebbles efficiently when it approaches 10−2 M⊕. Accounting for migration, close-in super-Earth planets form. Super-Earth planets are likely to form when the pebble mass flux is higher, the disk turbulence is lower, or the Stokes number of the pebbles is higher. Conclusions. For the single site planetesimal formation scenario, a two-component mass distribution with a large embryo and small planetesimals promotes planet growth, first by planetesimal accretion and then by pebble accretion of the most massive protoplanet. Planetesimal formation at single locations such as ice lines naturally leads to super-Earth planets by the combined mechanisms of planetesimal accretion and pebble accretion.

Accretion of the asteroids: Implications for their thermal evolution

AbstractThermal models of asteroids generally assume that they accreted either instantaneously or over an extended interval with a prescribed growth rate. It is conventionally assumed that the onset of accretion of chondrite parent bodies was delayed until a substantial fraction of the initial 26Al had decayed. However, this interval is not consistent with the early melting, and differentiation of parent bodies of iron meteorites. Formation time scales are tested by dynamical simulations of accretion from small primary planetesimals. Gravitational accretion yields rapid runaway growth of large planetary embryos until most smaller bodies are depleted. In a given simulation, all asteroid‐sized bodies have comparable growth times, regardless of size. For plausible parameters, growth times are shorter than the lifetime of 26Al, consistent with thermal models that assume instantaneous accretion. Rapid growth after planetesimal formation is consistent with differentiation of parent bodies of iron meteorites, but not with the assumed delay in formation of chondritic bodies. After the initial growth stage, there is an interval of slower evolution until the belt is stirred and the embryos are dynamically removed. During this interval, a fraction of asteroid‐sized bodies experience large accretional impacts, allowing bodies of the same final size to have very different histories of radius versus time. Accretion from small primary planetesimals leaves some fraction of material in bodies small enough to preserve CAIs while avoiding heating by 26Al. Unheated material can be a significant fraction of the mass that remains after large embryos are removed from the Main Belt.

Physics of planet trapping with applications to HL Tau

ABSTRACT We explore planet formation in the HL Tau disc and possible origins of the prominent gaps and rings observed by ALMA. We investigate whether dust gaps are caused by dynamically trapped planetary embryos at the ice lines of abundant volatiles. The global properties of the HL Tau disc (total mass and size) at its current age are used to constrain an evolving analytic disc model describing its temperature and density profiles. By performing a detailed analysis of the planet–disc interaction for a planet near the water ice line including a rigorous treatment of the dust opacity, we confirm that water is sufficiently abundant (1.5 × 10−4 molecules per H) to trap planets at its ice line due to an opacity transition. When the abundance of water is reduced by 50 ${{\ \rm per\ cent}}$ planet trapping disappears. We extend our analysis to other planet traps: the heat transition, dead zone edge, and the CO2 ice line and find similar trapping. The formation of planets via planetesimal accretion is computed for dynamically trapped embryos at the water ice line, dead zone, and heat transition. The end products orbit in the inner disc (R &amp;lt; 3 au), unresolved by ALMA, with masses that range between sub-Earth to 5 Jupiter masses. While we find that the dust gaps correspond well with the radial positions of the CO2, CH4, and CO ice lines, the planetesimal accretion rates at these radii are too small to build large embryos within 1 Myr.

Extension of the span and optimization of the optical "magic carpet": generation of a wide quasi-nondiffracting light sheet.

Light sheet illumination is the basis in developing light sheet microscopy (LSM), a technique with significant advantages compared with other classical techniques. Most proposed optical systems to generate light sheets for LSM use many optical elements, which require extensive adjustments and are costly; moreover, they generate a nonuniform or semiuniform light sheet or have a short depth of field (DOF). A simple scheme using a pair of double slits and a cylindrical lens for generating a quasi-nondiffracting 2D light sheet was reported in Opt. Lett.40, 5121 (2015)OPLEDP0146-959210.1364/OL.40.005121. In the present investigation, we elaborate on the optimization of the mask used. As the separation between the two slits increases, the light sheet becomes thinner and the DOF smaller and vice versa. The slits' width does not affect the light sheet thickness, but it does affect the intensity of the side lobes. For convergence angles of the inner slits from 0.75° to 8°, an optimum ratio of the slits' separation/width of 2.182 is recommended. The obtained light sheet is quasi-diffraction-free, namely, while its DOF is comparable with that of a Gaussian beam, its diffraction broadening is substantially smaller. We also add to the previously developed configuration a Powell lens in order to expand the beam in the spanwise direction while keeping nearly constant intensity in this dimension. We perform scalar diffraction theory calculations and conduct measurements showing the nearly constant intensity in the significantly broadened span of the light sheet. Potential applications for the augmented width include imaging of certain large embryos, laser micromachining, and microparticle image velocimetry.

Small day 8 equine embryos cannot be rescued by a less advanced recipient mare uterus

Equine embryos tolerate an unusually large degree of negative uterine asynchrony (recipient mare up to 5 days behind the donor mare). By contrast, positive asynchrony of more than 2 days results in a high incidence of early embryonic loss (EEL). Day 8 embryos range in diameter from approximately 130-1300 μm, with embryos smaller than 300 μm reported to suffer an increased incidence of EEL. However, it is not known whether this reduced viability is due to intrinsically poor embryo quality, or to inadvertent recipient uterine stage-embryo (positive) asynchrony. To examine whether small embryos survive better in Day 4–5 recipients than in recipients with a more advanced uterine stage, the likelihood of pregnancy (PR) and EEL for 62 small (<300 μm) and 215 larger Day 8 horse embryos were compared after transfer to recipients at different uterine stages (Days 4–5, 6–7 and 8–9) using logistic regression. Overall, EEL was higher (21.2%; P < 0.05) for small than larger embryos (7.1%). However, neither PR nor EEL were influenced by the recipient's uterine stage at the time of transfer (P > 0.1). The EEL for small embryos transferred into Day 4–5, 6–7 and 8–9 recipients was 20.8, 18.7 and 25.0%, respectively. We conclude that embryos recovered on Day 8 with a diameter <300 μm are at increased risk of EEL due to reasons other than inadvertent positive asynchrony with the recipient mare's uterus.

Gene expression in the phenotypically plastic Arctic charr (Salvelinus alpinus): A focus on growth and ossification at early stages of development.

Gene expression during development shapes the phenotypes of individuals. Although embryonic gene expression can have lasting effects on developmental trajectories, few studies consider the role of maternal effects, such as egg size, on gene expression. Using qPCR, we characterize relative expression of 14 growth and/or skeletal promoting genes across embryonic development in Arctic charr (Salvelinus alpinus). We test to what extent their relative expression is correlated with egg size and size at early life‐stages within the study population. We predict smaller individuals to have higher expression of growth and skeletal promoting genes, due to less maternal resources (i.e., yolk) and prioritization of energy toward ossification. We found expression levels to vary across developmental stages and only three genes (Mmp9, Star, and Sgk1) correlated with individual size at a given developmental stage. Contrary to our hypothesis, expression of Mmp9 and Star showed a non‐linear relationship with size (at post fertilization and hatching, respectively), whilst Sgk1 was higher in larger embryos at hatching. Interestingly, these genes are also associated with craniofacial divergence of Arctic charr morphs. Our results indicate that early life‐stage variation in gene expression, concomitant to maternal effects, can influence developmental plasticity and potentially the evolution of resource polymorphism in fishes.

THE INTRAPOPULATION HETEROSPERMIA OF SOME ORCHID SPECIES

The aim of the work was a comparative morphological study of the peculiarities of the structure of seeds of certain Orchidaceae species from various subfamilies growing on the territory of the mainland Ukraine and Crimea. Methods. In order to carry out the morphological analysis , the seeds were examined on a scanning electron microscope JSM-6060 LA (JEOL, Japan). The size of the structures on the obtained microphotographs was determined using the UTHSCSA Image Tool 3.0 program. The morphometric parameters of the seeds were determined in accordance with the average size of viable seeds for a particular species of orchids. The seed index , the embryo index , and the relative size of the embryo were analyzed. The results of the comparative study of features of the structure of seeds of orchids ( Orchidaceae family) of domestic flora: ( Cephalanthera longifolia (L.) Fritsch., Epipactis palustris (L.) Crantz, Dactylorhiza incarnata (L.) Soo. s.l., Anacamptis coriophora (L.) R.M. Bateman) are presented. The significant heterogeneity of seeds of the studied species is revealed. The morphological analysis established that at the time of dissemination the major part of the studied seeds of orchids were nonviable. Species of orchids with the greatest part of viable seeds with large embryos ( D. incarnata and C. longifolia ) are considered as perspective reintroducents. Conclusion . The results of the research can contribute to the scientific and practical developments aimed at creation of reserve genetic collections in botanical gardens and in reserved areas and also at restitution of a number of natural populations of rare and endangered species.

Influence of follicle size on bovine oocyte lipid composition, follicular metabolic and stress markers, embryo development and blastocyst lipid content.

This study assessed the lipid composition of oocytes from different follicle sizes and compared the expression of lipid-related genes and follicular fluid (FF) molecules between groups. We also investigated the functional consequences of differences on embryo development and blastocyst lipid deposits. Oocytes and FF were recovered from different follicle sizes. Oocytes from small (≤5mm) and large (≥6mm) bovine follicles were used to produce Day 7 expanded blastocysts (Day7Ex) and blastocysts that only became expanded at Day 8 (Day8Ex) after insemination. Oocytes from >8mm follicles had the highest lipid content. Few oocyte phospholipid variations were identified between groups. Very long chain fatty acid elongase 6 (ELOVL6) mRNA abundance was reduced in larger follicle-derived oocytes compared with the ≤2mm group. Increased levels of glucose, reactive oxygen species, glutathione and superoxide dismutase activity were also identified in FF from larger follicles. Large follicle-derived embryo development and lipid content of Day7Ex were greater than those derived from small follicles. Day8Ex had greater lipid deposition than Day7Ex. Oocytes and blastocysts exhibited follicle size-specific lipids. Large-follicle oocytes had increased lipid content and became Day7Ex with greater lipid deposition whereas delayed blastocoel expansion associated with a prolonged period of culture determined the lipid accumulation of Day8Ex. The FF microenvironment of large follicles seems to favour embryo development.

Rapid preimplantation genetic screening using a handheld, nanopore-based DNA sequencer

To determine if a handheld, nanopore-based DNA sequencer can be used for rapid preimplantation genetic screening (PGS). Laboratory study. Academic medical center. Amplified genomic DNA from euploid and aneuploid trophectoderm biopsy samples (n=9) that was also tested using traditional next generation sequencing (NGS). Short-read DNA library preparation and nanopore-based sequencing using a hand-held MinION sequencer. Comparison of cytogenetic testing result from NGS and nanopore-based sequencing and the time required for library preparation and sequencing. Multiplexed short-read DNA library preparation was completed in 45minutes. Sequencing on a single sample was completed within 20minutes and 5 samples were simultaneously sequenced in under 2hours. Whole-chromosome aneuploidy screening results obtained from nanopore-based sequencing were identical to those obtained using NGS. Here we report the first application of nanopore-based sequencing for PGS on trophectoderm biopsy samples using a novel rapid multiplxed short-read nanopore sequencing library preparation protocol. Sequencing for aneuploidy screening could be performed on a single sample in 20minutes and on 5 samples, simultaneously, within 2hours. Overall, nanopore sequencing is a promising tool to perform rapid PGS onsite, enabling same day testing and embryo transfer, thus obviating the need for complex, large and expensive DNA sequencers or embryo freezing.

A Dilution Model for Embryonic Scaling

A recent study in Nature Cell Biology (Almuedo-Castillo etal., 2018) describes a mechanism for tissue scaling in zebrafish embryos. The authors show that a fixed relative amount of the diffusible Nodal inhibitor Lefty produces an extended gradient in larger embryos, ensuring proportionally scaled germ layers, irrespective of embryo size.

Is aspiration of blastocoelic fluid essential to vitrify large equine embryos?

Is aspiration of blastocoelic fluid essential to vitrify large equine embryos?

Moonfalls: collisions between the Earth and its past moons

During the last stages of the terrestrial planet formation, planets grow mainly through giant-impacts with large planetary embryos. The Earth's Moon was suggested to form through one of these impacts. However, since the proto-Earth has experienced many giant-impacts, several moons are naturally expected to form through a sequence of multiple (including smaller scale) impacts. Each impact potentially forms a sub-Lunar mass moonlet that interacts gravitationally with the proto-Earth and possibly with previously-formed moonlets. Such interactions result in either moonlet-moonlet mergers, moonlet ejections or infall of moonlets on the Earth. The latter possibility, leading to low-velocity moonlet-Earth collisions is explored here for the first time. We make use of SPH simulations and consider a range of moonlet masses, collision impact-angles and initial proto-Earth rotation rates. We find that grazing/tidal-collisions are the most frequent and produce comparable fractions of accreted-material and debris. The latter typically clump in smaller moonlets that can potentially later interact with other moonlets. Other collision geometries are more rare. Head-on collisions do not produce much debris and are effectively perfect mergers. Intermediate impact angles result in debris mass-fractions in the range of 2-25% where most of the material is unbound. Retrograde collisions produce more debris than prograde collisions, whose fractions depend on the proto-Earth initial rotation rate. Moonfalls can slightly change the rotation-rate of the proto-Earth. Accreted moonfall material is highly localized, potentially explaining the isotopic heterogeneities in highly siderophile elements in terrestrial rocks, and possibly forming primordial super-continent topographic features. Our results can be used for simple scaling laws and applied to n-body studies of the formation of the Earth and Moon.

Is Aspiration of Blastocoelic Fluid Essential to Vitrify Large Equine Embryos?

Equine embryo vitrification is still not a well-established technique in equine practice. Notably, little work has been done on the effect of the warming system on viability of vitrified embryos. Our goal was to evaluate the effect of warming without cryoprotectants on in vitro - produced (IVP) embryo viability in culture, quality assessment parameters, and pregnancy after transfer. Equine IVP blastocysts were vitrified using commercial embryo vitrification media and a semi-closed vitrification device. In Exp. 1, we evaluated two warming temperatures (room temperature, RT, ∼22 °C; and 38 °C) for each of three warming systems: commercial warming solution (Kit); commercial embryo holding medium (EHM) with decreasing concentrations of sucrose (EHM + SS); or EHM alone without added sucrose. Embryos (n = 9 to 14 per treatment) were cultured in vitro for 24 h, stained with DAPI, TUNEL, and fluorophore-labelled phalloidin, and evaluated for nucleus number, mitotic rate, apoptotic rate, and actin filament distribution. In Exp. 2, to survey embryo viability in vivo, vitrified IVP blastocysts were shipped to an embryo transfer facility, then warmed immediately before transfer to recipient mares, using the warming treatments associated with the nominally best (Kit-RT, Kit-38, EHM-RT) and poorest (EHM + SS-38) assessed embryo quality in Exp. 1 (n = 7 to 8 per treatment). Subsequently, IVP blastocysts produced as part of our clinical program were vitrified and shipped, then warmed in embryo holding medium at an embryo transfer facility before transfer to recipient mares; fresh IVP embryos were shipped and transferred as controls. In Exp. 1, embryos increased significantly in diameter after culture (P < 0.01), with no difference among treatments. There was no difference (P > 0.05) in the number of viable nuclei, apoptotic rate, or microfilament distribution among treatments, or between vitrified-warmed and Control embryos. The mitotic rate was higher (P = 0.021) for Kit-RT (3.6%) when compared with the other treatment groups (1.5–2.0%). In Exp. 2, there was no difference (P > 0.05) in initial pregnancy (71.4–87.5%) or heartbeat (57.1%–85.7%) rates among warming treatments. In the clinical trial, there was no difference (P > 0.05) between vitrified-warmed and Control embryos in initial pregnancy (90.9% and 66.6%, respectively) or heartbeat (81.8% and 66.6%, respectively) rates. These results indicate that a semi-closed vitrification system using commercially-available media, and incorporating warming in the field in a single step using commercial embryo holding medium without cryoprotectants, can provide high pregnancy rates with IVP equine embryos.

Migration-driven diversity of super-Earth compositions

ABSTRACT A leading model for the origin of super-Earths proposes that planetary embryos migrate inward and pile up on close-in orbits. As large embryos are thought to preferentially form beyond the snowline, this naively predicts that most super-Earths should be very water-rich. Here we show that the shortest period planets formed in the migration model are often purely rocky. The inward migration of icy embryos through the terrestrial zone accelerates the growth of rocky planets via resonant shepherding. We illustrate this process with a simulation that provided a match to the Kepler-36 system of two planets on close orbits with very different densities. In the simulation, two super-Earths formed in a Kepler-36-like configuration; the inner planet was pure rock while the outer one was ice-rich. We conclude from a suite of simulations that the feeding zones of close-in super-Earths are likely to be broad and disconnected from their final orbital radii.

Improvements of doubled haploid production protocol for white cabbage (Brassica oleracea var. capitata L.)

Abstract Protocols leading to the development of doubled haploid (DH) lines by microspore culture are widely used in white cabbage (Brassica oleracea var. capitata L.), but efficiency varies according to the cultivar and induction procedure. Forty different genotypes consisting of F1 cultivars and their crosses with responsive doubled haploid lines were tested to evaluate the androgenic response. In total, 20,032 embryos were produced. On average, the haploid induction response of F1 cultivars was 7.0 embryos/Petri dish, but the average of these hybrids crossed to responsive DH lines was 26.6 embryos/Petri dish. In seven reciprocal crosses, a difference was observed in just one, meaning that the maternal effect probably has a minor influence on haploid embryogenesis in cabbage. Addition of 0.02% activated charcoal (AC) to the induction media increased embryo formation in several low-responsive genotypes, but its effect on embryo formation of high-responsive genotypes was predominantly negative, although larger embryos were formed on media containing AC than without AC. Further development into plantlets was tested by two procedures. Formed embryos were either transferred directly to regeneration medium or treated with abscisic acid and desiccated for 4 weeks. Regrowth and further development reached on average 15.5 and 57.6%, for the first and second procedures, respectively. Plantlets developed by direct transfer often exhibited abnormal development or hyperhydricity, unlike the desiccated embryos. Spontaneous diploidisation of embryos reached 42.5% in total and was not affected by AC added to the induction media.

A 3-dimensional (3D)-printed Template for High Throughput Zebrafish Embryo Arraying.

The zebrafish is a globally recognized fresh water organism frequently used in developmental biology, environmental toxicology, and human disease related research fields. Thanks to its unique features, including large fecundity, embryo translucency, rapid and simultaneous development, etc., zebrafish embryos are often used for large scale toxicity assessment of chemicals and drug/compound screening. A typical screening procedure involves adult zebrafish spawning, embryos selection, and arraying the embryos into multi-well plates. From there, embryos are subjected to exposure and the toxicity of chemical, or the effectiveness of the drugs/compounds can be evaluated relatively quickly based on phenotypic observations. Among these processes, embryos arraying is one of the most time-consuming and labor-intensive steps that limits the throughput level. In this protocol, we present an innovative approach that makes use of a 3D-printed arraying template coupled with vacuum manipulation to speed up this laborious step. The protocol herein describes the overall design of the arraying template, a detailed experimental setup and step-by-step procedure, followed by representative results. When implemented, this approach should prove beneficial in a variety of research applications using zebrafish embryos as testing subjects.

Barley lys3 mutants are unique amongst shrunken-endosperm mutants in having abnormally large embryos

Many shrunken endosperm mutants of barley (Hordeum vulgare L.) have been described and several of these are known to have lesions in starch biosynthesis genes. Here we confirm that one type of shrunken endosperm mutant, lys3 (so called because it was first identified as a high-lysine mutant) has an additional phenotype: as well as shrunken endosperm it also has enlarged embryos. The lys3 embryos have a dry weight that is 50–150% larger than normal. Observations of developing lys3 embryos suggest that they undergo a form of premature germination and the mature lys3 grains show reduced dormancy. In many respects, the phenotype of barley lys3 is similar to that of rice GIANT EMBRYO mutants (affected in the OsGE gene). However, the barley orthologue of OsGE is located on a different chromosome from Lys3. Together these results suggest that the gene underlying Lys3 is unlikely to encode a starch biosynthesis protein but rather a protein influencing grain development.