Histological characterization of a monoclonal antibody raised against the branchial arches of the chick embryo: reactivity with myogenic lineages and a few non-mesodermal derivatives

Summary Monoclonal antibody A7H2 has been selected from a library of monoclonals raised against the branchial arch cells of 3-day-old chicken embryos, and its histological distribution was examined at different embryonic stages and levels. The immunoreactivity appeared during neurulation and was almost general in extraembryonic areas. As the embryo formed, A7H2-fluorescence disappeared from most tissues, but was concentrated in the splanchnomesoderm-derived smooth muscle lineages and epithelial linings of the coelomic cavity. A strong and durable reactivity was also observed in the myogenic condensations constituting the axis of the branchial arches, whereas myotomal cells of the differentiating somites were labelled more weakly and for a shorter time. Interestingly, non-mesodermal fluorescence was restricted to the branchial arch epithelium and some ectodermal placodes, to the thinning-out zones of the neural tube destined to form the choroid plexus, and to the pharyngeal and cloacal extremities of the digestive tract.     

Soybean somatic embryogenesis: Effects of hormones and culture manipulations

Somatic embryos were induced in cultures of immature soybean (Glycine max (L.) Merr) embryos, or isolated cotyledons on MS modified medium supplemented with NAA and 2,4-D, BAP and ABA. When NAA and 2,4-D were compared at similar concentrations (25 and 23 M), 2,4-D produced larger number of somatic embryos, however, embryogenesis efficiency was improved in media containing NAA by using higher levels (100–150 M) of the auxin. Somatic embryo morphology varied with auxin type: NAA-induced embryos more closely resembled zygotic embryos than did 2,4-D-induced embryos. Additions of BAP or ABA to auxin-containing media had either no effect or reduced embryo production, although ABA altered the morphology of 2,4-D-induced embryos. In media containing both NAA and 2,4-D, the latter was dominant in terms of embryo morphology. The effects of subculture frequency and of transfers between 2,4-D and NAA media were investigated: Subculture frequency influenced mainly the frequency of normal embryos, while preculture on 2,4-D increased subsequent embryogenesis efficiency on NAA medium but reduced the frequency of normal embryos.Abbreviations Em^-2 s^-1 microEinsteins per square meter per second - NAA -naphthalene acetic acid - 2,4-D 2,4-dichlorophenoxy acetic acid - ABA abscisic acid - BAP benzylamino purine This paper (No. 86-3-96), is published with the approval of the director of the Kentucky Agricultural Experiment Station.     

Factors affecting adventitious bud induction in Pinus elliottii (Engelm.) embryos cultured in vitro

Embryos of slash pine (Pinus elliottii Engelm.) were induced to form adventitious buds when placed in culture on nutrient media supplemented with cytokinin. Buds were induced on media containing Risser & White major salts. The high content in nitrogen of Murashige & Skoog formulation seems to be deleterious for this in vitro system, since morphogenic responses were only promoted when nitrogen concentration was drastically reduced in the macronutrient formulation. Factors such as concentration of cytokinin (6-benzyladenine) and time and method of exposure (liquid or solid induction medium) strongly influenced bud formation and development. The greatest number of buds and shoots were obtained from 22.0 M cytokinin, but these shoots showed less and slower development than those induced with low dosages of cytokinin. The presence of naphthaleneacetic acid in combination with cytokinin in the induction medium decreased the frequency of bud formation.Abbreviations (BA) 6-benzyladenine - (NAA) 1-naphthaleneacetic acid     

Measurement of intra-embryonic pH during the early stages of development in the chick embryo

Summary Measurements have been made of the pH in the extracellular space, adjacent to the neural tube, in 73 isolated chick embryos in vitro at stages from 4–22 somites. A pH of 7.8–8.4 was observed in the segmented region, while caudally, in the segmental plate, the pH was consistently lower falling by as much as 0.5 pH units at the regressing primitive streak. Variations were noted in the pH of embryos of the same age but the regional variation in pH was a consistent finding in all of the embryos examined. The buffering capacity of the extracellular space was found to be 12.9 mequiv/pH unit/1 in the segmented region and 13.9 mequiv/pH unit/1 in the segmental plate. Thus it is unlikely that the regional variations in pH result from local variations in the buffering power of the extracellular space. Varying the K⁺, Cl^-, Mg²⁺ or HCO ₃ ^- ion concentrations in the bathing medium caused little change in the intra-embryonic pH, while reducing the concentrations of Na⁺ or Ca²⁺ caused a small acidification. This suggests that the ectoderm and endoderm form an effective barrier between the embryo and the external environment. Exposure of the embryo to KCN reduced the intra-embryonic pH suggesting that the alkaline environment is maintained by active processes.     

刺梨胚及胚乳的发育

本研究对单瓣刺梨胚及胚乳的发育过程进行了观察,获得如下主要结果:1.刺梨胚的发育属于紫菀型的一种变异类型。原胚发育早期,在胚体顶端具有明显的胚芽原细胞。成熟胚为典型的双子叶植物胚的形态,在子叶中贮藏大量的蛋白质粒。2.刺梨的胚乳属核型。经游离核时期以后形成胚乳细胞。紧邻胚囊周界壁的表层胚乳细胞可以进行平周分裂,产生层叠状的胚乳周缘层。此种后形成的胚乳,我们称之为次生胚乳。当次生胚乳形成时,其余的胚乳细胞逐渐解体,最后几乎完全消失。次生胚乳只在合点处解体,其余保留至种子成熟。3.发现了开花后一些胚珠中无胚或胚和胚乳在发育早期退化的现象,可认为是刺梨种子不育的一个重要原因。     

Endogenous β-d-galactoside binding lectin during the expansion of the yolk sac in the developing chick embryo

Summary A lectin with an affinity for -d-galactoside-containing saccharides is present in the developing yolk sac from the chick embryo at stages from 2 to 7 days of incubation. This activity is present in the area vitellina (less differentiated) and the area vasculosa (more differentiated). In both areas, lectin activity increases significantly during the spreading of the yolk sac up to 5 days of incubation. At all of the stages studied lectin activity was significantly higher in the area vasculosa, as compared to the area vitellina.Lectins were purified by affinity chromatography and examined by SDS-PAGE. Under reducing conditions two components are evident. A more prominent band of subunit molecular weight of 14,200±100 for the area vitellina and 13,700±300 for the area vasculosa and a second band with molecular weight of about 68,000±700 and 68,000±1,200 for the area vitellina and area vasculosa respectively, were observed. The -d-galactoside-binding lectin appears to be similar if not identical to that of the early chick blastoderm.     

Effect of ice nucleation by droplet of immobilized silver iodide on freezing of rabbit and bovine embryos

Silver iodide was immobilized by applying the insoluble reaction between sodium alginate and calcium chloride. The immobilized silver iodide was immersed into a freezing solution in order to trigger ice nucleation. Temperature change during cooling and postthaw in vitro development of embryos were examined in order to evaluate the effectiveness of the immobilized silver iodide (AgI alginate-gel droplet) on embryo development. Samples containing the AgI alginate-gel droplets released the latent heat of fusion at a higher subzero temperature than samples without the AgI alginate-gel droplets. When the AgI alginate-gel droplet was added to the freezing solution of rabbit and bovine embryos, they were successfully preserved in liquid nitrogen.     

A system for routine plantlet regeneration of sunflower (Helianthus annuus L.) from immature embryo-derived callus

Immature embryos from inbred commercial cultivars of sunflower (Helianthus annuus L.) were used as donor material for induction of regenerable tissue in vitro. Optimum regeneration frequencies were obtained by transferring the tissue through a sequence of defined media using a specified timetable. The first medium was characterized by a high sucrose content (12%), 10 M 2,4-dichlorophenoxyacetic acid and 5 M abscisic acid. Within 7 days, proliferation of smooth, white, dedifferentiated tissue from the cotyledons was evident. After 3 weeks, the tissues were transferred through a series of 3 media, designed to promote shoot formation, shoot elongation and rooting. Regenerates were obtained with all 7 genotypes tested. From 1983 to 1984, approximately 500 primary R₀ plantlets were regenerated, grown to maturity in a greenhouse and self-pollinated. The resultant R₁ seeds were subsequently field-grown and the plants were evaluated for variation.     

Parasitism of aphid embryos by Aphidius smithi: some effects of extremely small host size

Embryos of the pea aphid, Acyrthosiphon pisum (Harris), were successfully parasitized by Aphidius smithi Sharma & Subba Rao while still inside their mother. Parasites from embryos were much smaller, laid fewer eggs, but survived as long as their counterparts that had developed in adult viviparous aphids. Lifetime fecundity was positively correlated with wasp size, as measured by dry weight at the time of death. A. smithi females produced the same offspring sex ratio at eclosion on both host classes, although emergence was significantly lower from mummified adult aphids than from embryos. The data are discussed with regard to progeny and sex allocation decisions by solitary hymenopterous parasites. It is suggested that, in A. smithi, the decision to lay a fertilized or unfertilized egg is influenced by the host's overall size, rather than by the size and quality of a smaller target area inside the host, such as embryos.     

Effects of growth regulator concentrations and explant size on shoot organogenesis from callus derived from zygotic embryos of sunflower (Helianthus annuus L.)

Effects of medium growth regulator composition and embryo size on shoot organogenesis of callus derived from globular- to torpedo-shaped zygotic embryos of five sunflower (Helianthus annuus L.) genotypes were examined. Forty growth regulator combinations composed of 0 to 5 mgl^-1 naphthaleneacetic acid (NAA) and 0 to 1 mgl^-1 6-benzylaminopurine (BA) were tested. The frequency of zygotic embryos forming shoot-regenerating callus was analysed according to categorical data modelling using a maximum-likelihood approach. Both NAA and BA must be present to induce the formation of morphogenic callus from zygotic embryos, but each growth regulator effect varied with the genotype. For four genotypes, NAA and BA effects were neither linear nor quadratic; whereas, they were linear for the fifth one. Most effective concentrations across genotypes were 0.1 mgl^-1 NAA and 0.5 mgl^-1 or 0.2 mgl^-1 BA. However, the optimal growth regulator combination depended on the genotype and an interaction between the two growth regulators. The frequency of shoot-regenerating callus also varied with the size of the embryo explant. For all five genotypes, 0.4 to 1.2 mm long heart-shaped zygotic embryos formed morphogenic callus more frequently than smaller less-developed ones.