Hypoxylon cohaerens: Cytology of the ascus

The haploid chromosome number ofHypoxylon cohaerens apparently is 5, based on counts made at meiotic prophase and meiotic and mitotic metaphases. Newly formed ascospores are at first uninucleate, becoming binucleate following mitosis in the ascospore. Subsequently, one of the two nuclei disappears. Maturing ascospores are uninucleate.Scientific Paper No. 3732 Washington State University. College of Agriculture, Project 1767. This study was supported in part by National Science Foundation Grant GB-19924.     

Chromosome number in the lizard genus Uta (family Iguanidae)

A chromosome number of 34 (12 macro- and 22 microchromosomes) was found to be characteristic of the bone marrow in 47 animals including males from the species Uta antiquus, and both males and females from the following species and subspecies: Uta stansburiana stansburiana, Uta stansburiana stejnegeri, Uta stansburiana elegans, Uta stansburiana klauberi, Uta stansburiana mannophorus, Uta nolascensis, Uta palmeri, and Uta squamata. — Diploid chromosome numbers of 34 and haploid numbers of 17 were found in the nine testis smears examined. — The presence of a large number of hypodiploid figures in the bone marrow smears is attributed to cell fragmentation and the problem of distinguishing the small microchromosomes. — Series of polyploid figures whose chromosome numbers increased in arithmetic rather than geometric progressions were observed in the testis dry smears. Possible alternatives for the origin of these figures are presented. — Problems encountered in the use of chromosome number as a taxonomic character are discussed.Supported in part by Research Grants GB-366 and GB-5416 from the National Science Foundation, and GM-15361 from the United States Public Health Service.     

A specific chromosome element,the telomere of Drosophila polytene chromosomes

The submicroscopic organization of terminal chromosome regions of Drosophila hydei polytene chromosomes is described. A compact region composed of tightly packed fibrils of 100 to 125 Å diameter embedded in an amorphous material is located at each of the chromosome ends of the 5 long chromosome arms. From this compact region, sometimes containing cavities, fibrils extend onto the nearest normal band region. The diameter of the extending fibrils is 100–125 Å, 200–250 Å or 400 Å. Pronase digestion of fixed and squashed chromosomes reduced the electron density of the amorphous matrix in the compact regions but failed to affect the diameter of the fibrils. The extending fibrils, however, showed a decrease in diameter after pronase digestion. The most frequently observed diameter values were 100–125 Å. — The volume of the terminal structures, including the compact region as well as the extending fibrils, is characteristically different for the various elements of the karyotype. Chromosome 2 displays the largest terminal structure, whereas chromosome 4 only occasionally shows the presence of compact regions. — End to end association of the long chromosome arms involves the fusion of the compact terminal structures. The non-random distribution of end to end association seems to be correlated with the volume of the terminal structures. Chromosome 2 which contains the largest compact terminal region is more frequently involved in end to end associations than any other chromosome arm. — The terminal regions show replication of DNA. They belong to the group of regions which display a discontinuous labeling pattern along the chromosomes, representing a late phase of the replication cycle. — The unique structural organization of the terminal chromosome regions, which is never observed at any other location of the genome supports the idea that they are morphological manifestations of the postulated telomeres.     

Hypoxylon deustum: The chromosome number

Summary The haploid chromosome number ofHypoxylon deustum is 4. Ascospores are uninucleate when formed, becoming binucleate shortly thereafter. One of the two nuclei disappears in maturing ascospores. Mature ascospores are apparently uninucleate. The significance of these and other findings to the taxonomy of the fungus is discussed.Scientific Paper No. 3008. College of Agriculture, Washington State University. Project 3767.This study was supported in part by National Science Foundation Grant GB-5219.     

CHROMOSOME NUMBERS IN THE HYPOCREALES. II. ASCUS DEVELOPMENT IN NECTRIA CINNABARINA

The cytology of ascus development in Nectria cinnabarina was investigated with the orceinsmear technique, from crozier formation to ascospore maturation. At prophase I synapsis occurs while the chromosomes are still contracted, and the nucleus passes through dictyotene, a diffuse stage rarely seen in plants. A haploid complement of five chromosomes has been precisely determined. The first two divisions in the ascus constitute meiosis, and the third (mitotic) is followed by ascospore delimitation. A fourth division takes place in the ascospore, which is subsequently divided by a septum into two uninucleate cells. Of all species of Nectria thus far investigated N. cinnabarina is the only species in which additional nuclear divisions in the ascus do occur, accounting for the multinucleate condition in the ascospore cells. The bearing that this distinctive nuclear condition has on phylogeny and evolution in the Hypocreales is discussed.     

Cucullosporella mangrovei, ultrastructure of ascospores and their appendages

The ultrastructure ofCucullosporella mangrovei ascospores is described. Mature ascospores possess two wall layers, an outer electron-dense episporium and an innermost tripartite mesosporium. Episporial elaborations form electrondense spore wall ornamentations from which extend fibrils that may constitute a highly hydrated exosporium which was not visualised at either the scanning electron microscope or light microscope level. Ascospores possess a hamate appendage at each pole which unfolds in seawater to form a long thread. Ultrastructurally the polar appendage comprises folded fibro-granular electron-dense material and fine fibrils. The fibrils form a matrix around and within the fibro-granular appendage and around the entire unreleased ascospore. These fibrils have not been observed associated with the ascospore appendages in other species of the Halosphaeriales and are a discrete and new appendage component. The fibro-granular appendage and fibrils are bounded by the outer delimiting membrane which is absent around released ascospores. The nature of the spore appendage is compared with that of other marine and freshwater ascomycetes and the taxonomic assignment of the species is discussed.     

Taxonomic status of the species of the green algal genusNeochloris

Komárek has recently reviewed the various species assigned to the green algal genusNeochloris Starr (Chlorococcales, Chlorococcaceae) and removed those with uninucleate vegetative cells to a new genus,Ettlia. Watanabe & Floyd, unaware ofKomárek's work, also reviewed the species ofNeochloris and distributed them among three genera—Neochloris, Chlorococcopsis gen. nov., andParietochloris gen. nov.—on the basis of details of the covering of the zoospore and the arrangement of the basal bodies of the flagellar apparatus. This paper reconciles these two treatments and makes additional recommendations at the ranks of genus, family, order, and class.     

Wall texture in the spore of a vesicular-arbuscular mycorrhizal fungus

Summary InGlomus epigaeum Daniels and Trappe, a vesicular-arbuscular mycorrhizal fungus, the mature spore has a complex multi-layered wall containing a regular pattern of wall subunits.The outer wall (2–4 m thick) consists of a simple layer of parallel microfibrils. The inner wall (5–6 m thick) is built from two layers possessing different organization. The innermost layer, near the plasmalemma has a texture of apparently dispersed fibrils, whereas the second layer is regularly organized with an arced texture. Ten to twelve bundles of fibrils connected by apparently bow-shaped fibrils are consistently observed. The appearance of this arced organization depends on the section plane and on the angle of observation in the electron microscope as confirmed by tilting experiments. Wall subunits are evident as straight electron transparent fibrils; particularly well-defined in negatively stained frozen sections: their diameter is about 3.5nm.The regular pattern of wall subunits in this fungal cell wall is compared with the textures shown by cellulose fibrils in algae or higher plants and by chitin fibrils in arthropod cuticle.Research work supported by CNR, Italy. Special grant I.P.R.A.—Sub-project 1. Paper No. 55.     

Experimental alterations in the chromosome constitution of Sciara

Summary Using two reciprocal translocations between chromosomes X and IV in S. coprophila it has been possible to derive two kinds of aneuploid females. Both of the aneuploid complements are detrimental — one is lethal, the other may give rise to viable, fertile adults. Males with aneuploid somatic complements have not been obtained; three different aneuploid complements were tested but gave negative results. Males with euploid soma and aneuploid germ line have been produced in three separate instances; they are viable and fertile.Dedicated to Professor H. Bauer on the occasion of his sixtieth birthday.The studies reported here were supported by grant GB-42 from the National Science Foundation.     

Number and pattern of association of chromonemata in the chromosomes of Tradescantia

Summary Analysis of reconstructions, prepared from electron micrographs of successive longitudinal serial sections, has led to the conclusion that the somatic telophase chromosome of Tradescantia paludosa contains four cytologically separable chromonemata. The four represent a pair of pairs, that is, two diplospiremes — one with its two chromonemata arranged helically in dextrorse relationship, and the other with its two in sinistrorse relationship — which are associated to form a tetraspireme. During anaphase and telophase the tetraspireme constitutes the chromosome; during prophase and metaphase the tetraspireme represents one of the two chromatids of the chromosome, which is accordingly an octospireme in terms of the number of cytologically identifiable chromonemata. Loose intertwining of the two tetraspiremes during late prophase accounts for the so-called relational coiling.This paper is dedicated to Professor Hans Bauer on his sixtieth birthday anniversary in appreciation of his contributions to the development of modern cytology.The work reported here was supported in part by Research Grants GM-10499 from the National Institutes of Health, U.S. Public Health Service, and GB-290 from the National Science Foundation, and in part by a NATO fellowship awarded to E. Sparvoli by the Italian National Council of Research.     

Pleistophora finisterrensis n. sp., a microsporidian parasite of blue whiting Micromesistius poutassou

Pleistophora finisterrensis n. sp. is a microsporidian parasite of the hypoaxial musculature of the blue whiting Micromesistius poutassou (Risso). Foci of infection are between 3 and 6 mm in length and have no evident effects on adjacent muscle fibres. We found only a single type of spore (uninucleate, with mean dimensions of 4×2 µm in fresh preparations), contained within sporophorous vesicles (mean diameter 19 µm in fresh preparations; 150–250 spores per vesicle). All of the development stages of this microsporidian are monokaryotic. The meronts are initially uninucleate and bounded by a plasmalemma. Towards the end of merogony, meronts are multinucleate plasmodia with a well-defined surface coat. Sporogony is polysporous, with multinucleate sporonts, which likewise have a well-defined surface coat (about 130 nm thick), dividing by plasmotomy to give rise to uninucleate sporoblasts. The polar tube is isofilar and consists of 8–9 turns in the posterior half of spore. The polaroplast is made up of an anterior lamellar part and a posterior vesicular part.     

Ultrastructure of ascospore delimitation in freeze substituted samples ofAscodesmis nigricans (Pezizales)

Summary Freeze substitution proved to be a valuable technique for studying the early stages of ascosporogenesis inAscodesmis nigricans. Our observations indicate that the ascus vesicle originated from the ascus plasma membrane. Invaginations of the plasma membrane produced ascus vesicle initials consisting of two closely spaced unit membranes. The appearance of the outer leaflet of each of these membranes was identical to that of the inner leaflet of the ascus plasma membrane. Apparent points of continuity between ascus vesicle initials and the plasma membrane were observed. Ascus vesicle initials accumulated in the ascus cytoplasm near the plasma membrane and then coalesced to form the ascus vesicle, a peripheral, cylinder-like structure consisting of two closely spaced unit membranes that extended from the ascus apex to the ascus base. The ascus vesicle then became invaginated in a number of regions and subsequently gave rise to eight sheet-like segments, or ascosporedelimiting membranes, that encircled uninucleate segments of cytoplasm forming ascospore initials. Like the ascus vesicle, each ascospore-delimiting membrane consisted of two closely spaced unit membranes, the inner of which became the ascospore plasma membrane. The ascospore wall then developed between the spore plasma membrane and the outer membrane. Many details of ascospore maturation were clearly visible in freeze substituted samples.     

Electron histochemical study of 1:2-glycol groups in experimental hamster amyloid

Summary Leishmania-induced amyloid of hamster kidneys was studied by the periodic acid — thiocarbohydrazide — osmium tetroxide method which is analogous to the PAS reaction.The amyloid fibrils failed to give a reaction for 12-glycol groups. The ultrastructural distribution of mucosubstances containing 12-glycol groups was found to be localized in the ground substance between the fibrils.The reaction was unaffected by -amylase digestion, and chloroform-methanol extraction of lipids; it was dependant upon periodic acid oxidation.     

The inhibition of protein synthesis in meiotic cells and its effect on chromosome behavior

Autoradiographs show that tritiated leucine is incorporated into protein continually at an almost linear rate during meiotic prophase of lily microsporocytes in in vitro culture. Although label is mostly in the cytoplasm for the first hour, it becomes almost evenly distributed throughout the cell after a few hours. The amount of label decreases slightly, if at all, during a chase period extending through the rest of the prophase — a period of 3 to 4 days. — The incorporation of label was blocked by 95% by the protein inhibitor, cycloheximide, at a concentration of 3.5 × 10^-6 M. In the presence of this inhibitor, meiosis was arrested at all stages through metaphase I and even later. After temporary inhibition, however, or in low drug concentrations, characteristic cytological abnormalities subsequently developed, depending on the meiotic stage at which the inhibition occurred. One important observation was that the formation of chiasmata between homologs could be blocked if the inhibition was applied during the late zygotene or early pachytene stages.This work was supported by a grant from the National Science Foundation (GB-5173 X).USPHS postdoctoral fellow.     

Redheadia quercus gen. et sp. nov., the teleomorph of Mycopappus quercus, the frosty mildew fungus in Quercus acutissima

The teleomorph of Mycopappus quercus causing frosty mildew in Quercus acutissima is described as a new genus and species, Redheadia quercus, in the Sclerotiniaceae. Apothecia sprout from sclerotia on the fallen infected leaves kept for 10 months at 5°C and subsequent incubation at 15°C under diffused room light. Typical zonate lesions and multicellular propagules of M. quercus are produced on Q. acutissima, by mycelial inoculation using an isolate from a single ascospore, confirming the teleomorph–anamorphic connection. No significant differences are observed between cultured colonies of isolates from the ascospore and those from the propagule. Sclerotia and microconidia of the fungus are produced on culture media.     

Generation time and albumin evolution

The albumins of long-generation animals such as man and chimpanzee are shown to be no less evolved immunologically than those of some short generation time rodents such as rat, mouse, and hamster. This finding casts very serious doubt on certain assumptions concerning the times of divergence of these species made by DNA hybridization workers. As these assumptions have led them to conclude that generation time is a key factor controlling the rate of genomic or DNA evolution, this latter conclusion is also made suspect. The molecular clock concept—that it is absolute time which determines the amount of protein and DNA evolution—is, on the other hand, strengthened.This research was supported by NSF grants GB-20750 to the author and GB-13119 to Dr. A. C. Wilson.     

Studies on the fine structure of the dorsal vessel of arthropods

Summary The beetle family Altioidae contains some very archaic surviving species, and at the same time, very modern ones. In one lineage from archaic to modern (Fornterita — Alticini — Oedionychini), the number of spermatozoa has been stepwise (according to a geometric series, 2ⁿ) reduced from 256 to 16 per bundle. Comparison with the relatively few data in literature show that this trend is common in insects: more archaic orders have more sperm cells per bundle than the more modern ones, and within an order, the most modern or most specialized groups tend to have least. The reduced number of spermatozoa per bundle may be a sign of reduced sperm production in general, and this probably has adaptive value in limiting genetic variability.A small number of spermatozoa per bundle is sometimes accompanied by an extra large size of spermatocytes. Except for the alticid subtribe Oedionychina, it is hard to see what adaptive value this could have. In the Oedionychines, the large size of the spermatocytes is a prerequisition for the evolution of gigantism of the asynaptic sex chromosomes.Partly supported by a grant (GB-4522) from the National Science Foundation (Washington).I wish to express my gratitude to Prof. Dr. Jan Bechyné, Instituto de Zoología Agrícola, Universidad Central de Venezuela, Maracay, Venezuela, for identification of most alticids mentioned in this paper.     

The normal fine structure and aging changes of the human zonular fibers

Summary The normal zonular fibrils of the human eye do not differ from the fibrils of the zonula Zinnii of the rat. Furthermore, there is no difference between the single zonular fibrils and the fibrils of the vitreous body in rat and man. The average diameter of the human zonular fibrils is 109 Å. They are transversely striated at intervals with a periodicity of 70–150 Å. Over periods of mostly 400–440 å, but also of 500–640 Å were observed. At a few places over periods similar to those of the long spacing-type packing were found. Like in the rat eye, the zonular fibrils of the human eye must be regarded as a special form of collagen.An indirect proof for the collageneous nature of the zonular fibrils is the occurence in advanced age of degenerative alterations which are exclusively observed in connective tissue (hyalinization, elastoid degeneration).Supported by the Deutsche Forschungsgemeinschaft. Acknowledgement: I wish to thank Dr. H. Faßl, Institut für Medizinische Statistik und Dokumentation der Universität Mainz, for the statistical analysis.     

Structural aspects of amitosis: a light and electron microscope study of the isolated macronuclei of Paramecium aurelia and Tetrahymena pyriformis

Thin sections show the macronuclei of Paramecium aurelia and Tetrahymena pyriformis to contain two types of bodies. The smaller, measuring 0.1–0.2 in diameter, have been resolved in the light microscope by first removing the macronuclei from the cells in the presence of Mg⁺⁺, then chelating that divalent cation with EDTA, resulting in expansion of the nuclear material. By staining with methyl green, Azure B, and the Feulgen procedure, the small bodies were shown to contain DNA. In whole mounts these small bodies appear to be joined to one another producing a complex network suspended in which are the larger bodies, or nucleoli. — Macronuclei from both ciliates were isolated in large quantities and purified for spreading on an air-water interface. When the nuclei burst from surface tension forces and are examined with the electron microscope, the DNA containing bodies remain attached to one another by means of 100 Å fibrils. The pattern of attachment is non-linear. Occasionally individual DNA-containing bodies loosen revealing a coil resembling both in shape and dimensions the 250 Å coil characteristic of eucaryotic chromatin. The substructure of the 250 Å coil has not been directly observed. However, the frequent association of pairs of 100 Å fibrils makes it likely that two such fibrils are tightly complexed in the 250 Å coil. The 100 Å fibril, in turn, contains two 20 Å strands, each presumably a DNA double helix. — In Paramecium each small body of the macronucleus contains approximately one chromosome-equivalent of DNA. The fact that these small bodies are joined to form large structured masses of chromatin within the macronucleus indicates that the distribution of genetic material is not random. It is possible that, similar to bacteria, entire genomes segregate as units, thus accounting for successful amitotic division.This work was supported by a Predoctoral Fellowship to the author from the National Institutes of Health, U.S. Public Health Service, and by grant GM-13882 (NIH-USPHS) to Dr. Daniel Mazia.     

The structure of the chromosomes in human primordial oocytes

Primordial oocytes (oocytes in primordial follicles) from human ovaries aged 51/2 months post conception to 11 3/4 years post partum were examined in: (a) squash preparations of fresh and fixed tissue; (b) histological preparations; and (c) thin sections by electron microscopy, in order to study the structure of the chromosomes. — The light microscope shows that the chromosome consists of a thread bearing numerous fine lateral appendages. Cytochemical tests indicate that the thread contains DNA, and is surrounded by material containing RNA and protein. — The electron microscope shows that there are three main structural components in the chromosome: (i) an axis or core containing at least two longitudinal strands about 200 Å thick; (ii) a surrounding sheath composed of coiled fibrils which form symmetrically arranged columns and loops, and (iii) clusters of large granules which are associated with the outer parts of the sheath. Small nucleoli and other granular bodies are also present. — These observations indicate the presence of lampbrush chromosomes in the human oocyte. The significance of this type of chromosome in mammals is discussed in relation to the differential radiosensitivity of the oocytes, and to the form of chromosomes at the dictyate stage in rodents.