Studies on freshwater red algae of Malaysia III. development of carposporophytes ofBatrachospermum cayennense Montagne,B. beraense Kumano andB. hypogynum Kumano et Ratnasabapathy

The development of the carposporophytes ofBatrachospermum cayennense, B. beraense andB. hypogynum is observed. The carpogonium bearing branch ofB. cayennense is very long and its cells are differentiated from those of the vegetative fascicles in size and shape. The trichogyne nucleus ofB. cayennense andB. beraense is not recognized. In the above two species it is observed that after contact with the trichogyne the nucleus of the spermatium divides into two daughter nuclei, one of which fuses with the female nucleus, while the other remains in the trichogyne. InB. beraense the fertilized carpogonium is divided transversely and the gonimoblast filaments are produced from the divided carpogonium. The protoplasmic connection between the carpogonium and the hypogynous nutritive cell is especially pronounced inB. hypogynum. Carpospores ofB. hypogynum germinate within the carposporangia and young germlings often throw away original carpospores, hence these young germlings look like gemmae.     

Seedling establishment ofCastanopsis cuspidata var.sieboldii andPersea thunbergii on lava and scoria of the 1962 eruption on Miyake-jima Island,the Izu Islands

Seedling establishment ofCastanopsis cuspidata var.sieboldii Nakai andPersea thunbergii Kosterm. (Machilus thunbergii Sieb. et Zucc.) in early succession was studied on the lava and the scoria of the 1962 eruption on Miyake-jima Island of the Izu Islands. While tall individuals (> 1 m) ofP. thunbergii were found on both the lava and the scoria, those ofC. cuspidata (> 1 m) were found only on the lava. The height growth rate ofC. cuspidata was very low on the scoria, and there was no individual ofC. cuspidata taller than 0.4 m on the scoria. The proportion of yellow leaved individuals was much higher (37.1%) inC. cuspidata than inP. thunbergii (4.0%) on the scoria, while it was similar between the two species on the lava.Persea thunbergii seemed to be able to establish on both the lava and the scoria, butC. cuspidata established only on the lava. The differences in growth rate and vigor between the two species on the lava and the scoria in early succession may influence later stages of succession.     

Observations On Nuclear Division in vegetative hyphae of aquatic fungi

Summary Achlya aquatica, A. prolifera andA. racemosa are additional examples of fungi in whose vegetative mycelium, the nuclei do not divide in a manner directly comparable to ordinary mitosis. During division the entire nucleus, which consists of variously shaped chromatin and the nucleolus, becomes angular and elongates. The chromatin separates into two portions, which are situated at opposite ends of the elongating nucleus. As division proceeds the nuclei separate. Each sister nucleus sonsists of chromatin and a portion of the divided nucleolus. Spindless, chromosomal filaments or metaphase plates could not be observed during division.     

Meiotic differences between three triatomine species (Hemiptera,Reduviidae)

We have found the following differences in the male meiosis among three triatomine species: (1) The three largest autosomal bivalents ofTriatoma infestans are heterochromatic.Rhodnius prolixus has two autosomal bivalents with heterochromatic blocks.Triatoma rubrovaria does not show any heteropycnotic autosomes. (2) Sex chromosomes inT. infestans form a chromocenter. At early prophase terminal associations are seen between sex chromosomes inT. rubrovaria, and they maintain a close association until diakinesis. An intimate association between the X and Y chromosomes is observed during early prophase inR. prolixus, but a distant association is maintained by the sex chromosomes at diffuse and diplotene stages in this species. (3) Polyploid nuclei of the nutritive cells are quite distinct. Numerous chromocenters of different shapes and sized are seen in those ofT. infestans. InT. rubrovaria one chromocenter having two positively heteropycnotic elements is observed surrounded by homogeneous chromatin. Only one compact chromocenter is found amongst unevenly distributed chromatin, inR. prolixus.     

Chromosome banding in the genusPinus

The chromosomes (2n=24) ofPinus densiflora Sieb. et Zucc. andP. thunbergii Parl. collected from several localities were analyzed on their fluorescent banding patterns by sequential staining with the base specifically binding fluorochromes, CMA and DAPI. In both species, the CMA-bands were localized at the proximal and/or interstitial regions of most of the chromosomes. The CMA-banding pattern was constant among the cells in a plant and was specific to respective species with a few variations. After the CMA and DAPI stainings each chromosome was identified individually. The fluorescent banding patterns of the two species were somewhat similar, but were diferent with respect to in some characters.Pinus thunbergii had two pairs of metacentric chromosomes without CMA-band and two pairs of metacentric chromosomes with an additional thin CMA-band at the interstitial region. The 10th and 11th pairs of chromosomes of both species, which showed similarity in interstitial CMA and DAPI banding and chromosome shape, had the proximal CMA-bands inP. densiflora and DAPI-band inP. thunbergii. The interspecific F₁ hybrid between the two species could easily be identified by the fluorescent banding method.     

A light and electron microscopic study of mitosis inBullera alba and the histochemistry of some cytoplasmic substances

Summary Mitosis in the imperfect yeast-like basidiomyceteBullera alba was studied by comparative light and electron microscopy. During mitosis the chromatin containing part of the nucleus moved into the progeny cell, and the nucleolus containing part of the nucleus remained in the parent cell. The two portions of the nucleus then separated and the nucleolar part degenerated. Metaphase and anaphase took place in the progeny cell. Subsequently one mass of chromatin returned to the parent cell, and two new nuclei were formed. The study concentrated on the nuclear envelope, nucleolus, spindle pole body, chromatin, spindle, and cytoplasmic microtubules. Mitosis inB. alba was compared with reports of mitosis in other basidiomycetes, theUredinales, and theAscomycotina and was deemed closest to the heterobasidiomycete yeasts.Histochemical evidence for the presence of lipid, glycogen, and polyphosphate in the cytoplasm was presented.     

Functional role of newly formed pore complexes in postmitotic nuclear reorganization

Many nuclear proteins are released into the cytoplasm at prometaphase and are transported back into the daughter nuclei at the end of mitosis. To determine the role of this reentry in nuclear remodelling during early interphase, we experimentally manipulated nuclear protein uptake in dividing cells. Recently we and others have shown that signal-dependent, pore complex-mediated uptake of nuclear protein is blocked in living cells on microinjection of the lectin wheat germ agglutinin (WGA), or of antibodies such as PI1 that are directed against WGA-binding pore complex glycoproteins. In the present study, we microinjected mitotic PtK₂ cells with WGA or antibody PI1 and followed nuclear reorganization of the daughter cells by immunofluorescence and electron microscopy. The inhibitory effect on nuclear protein uptake was monitored by co-injection of the karyophilic protein nucleoplasmin. When injected by itself early in mitosis, nucleoplasmin became sequestered into the daughter nuclei as they entered telophase. In contrast, nucleoplasmin was excluded from the daughter nuclei in the presence of WGA or antibody PI1. Although PtK₂ cells with blocked nuclear protein uptake completed cytokinesis, their nuclei showed a telophaselike organization characterized by highly condensed chromatin surrounded by a nuclear envelope containing a few pore complexes. These findings suggest that pore complexes become functional as early as telophase, in close coincidence with nuclear envelope reformation. They further indicate that the extensive structural rearrangement of the nucleus during the telophase-G1 transition is dependent on the influx of karyophilic proteins from the cytoplasm through the pore complexes, and is not due solely to chromosome-associated components.Abbreviations WGA wheat germ agglutinin - GlcNAc N-acetylglucosamine     

Ultrastructural features of mitosis inLeishmania adleri

Summary The interphase nucleus ofLeishmania adleri has clumps of chromatin associated with the nuclear envelope and a large centrally located nucleolus. Prior to mitosis the basal bodies replicate at the cell anterior. Subsequently, dense plaques appear in the equatorial region of the nucleus at the time of spindle development. Microtubules appear in the nucleus adjacent to the nuclear envelope and embedded in the matrix of the plaques. A central spindle composed of a single bundle of microtubules develops and spans the nucleus. Plaques and nucleolar components laterally associate with the spindle and migrate towards the poles. The central spindle elongates to three to four times its original length separating the forming daughter nuclei and producing an interzonal spindle. A remnant of the interzonal spindle remains attached to each of the daughter nuclei until late into cytokinesis. The kinetoplast does not divide until after the completion of mitosis.     

An immunofluorescence study of mitosis in a mite-pathogen,Neozygites sp. (Zygomycetes: Entomophthorales)

Summary Mitosis in a mite-pathogenic species ofNeozygites (Zygomycetes: Entomophthorales) was investigated by indirect immunofluorescence microscopy using an antibody against -tubulin for visualization of microtubules (MTs). DAPI and rhodamine-conjugated phalloidin were used to stain chromatin and actin, respectively. Salient features of mitosis inNeozygites sp. are (1) a strong tendency for mitotic synchrony in any given cell, (2) conical protrusions at the poles of metaphase and anaphase nuclei revealed by actin staining, (3) absence of astral and other cytoplasmic MTs, (4) a spindle that occupies most of the nuclear volume at metaphase, (5) a spindle that remains symmetrical throughout most of mitosis, (6) kinetochore MTs that shorten during anaphase A, (7) a central spindle that elongates during anaphase B, pushing the daughter nuclei into the cell apices, and (8) interpolar MTs that continue to elongate even after separation of the daughter nuclei. Cortical cytoplasmic MTs are present in a few interphasic and post-cytokinetic cells. The data presented show thatNeozygites possesses features unique to this genus and support the erection of theNeozygitaceae as a separate family in theEntomophthorales.Abbreviations DAPI 4,6-diamidino-2-phenylindole - MT microtubule - SPB spindle pole body     

The relation between cell size,chromosome length and the orientation of chromosomes in dividing root cortex cells

Cells in the root meristem are organised in longitudinal files. Repeated transverse cell divisions in these files are the prime cause of root growth. Because of the orientation of the cell divisions, we expected to find mitoses with an spindle axis parallel to the file axis. However, we observed in the root cortex ofVicia faba large number of oblique chromosome orientations. From metaphase to telophase there was a dramatic increase of the rotation of the spindle axis. Measurements of both the size of the cortex cells and the chromosome configurations indicated that most cells were too small for an orientation of the spindle parallel to the file axis. Space limitation force the spindle into an oblique position. Despite this spindle axis rotation, most daughter cells remained within the original cell file. Only in extremely flat cells did the position of the daughter nuclei forced the cell to set a plane of division parallel to the file axis, which result in side-by-side orientation of the daughter cells. Telophase spindle axis rotations are also observed inCrepis capillaris andPetunia hybrida.. These species have respectively medium and small sized chromosomes compared toVicia. Since space limitation, which causes the rotation, depends both on cell and chromosome size, the frequency and extent of the phenomenon in former two species is comparatively low.     

Meiotic division and fusion of nuclei in multinucleate cells from the induced fusion of meiotic protoplasts of liliaceous plants

Multinucleate protoplasts were produced from meiotic cells at the zygotene and pachytene stages in a lily andTrillium, and their meiotic divisions were followed during subsequent culture. In each multinucleate, a complete synchrony of nuclear division was maintained throughout the meiotic process, and chromosome behavior appeared normal up to the metaphase stage. In most dinucleates, chromosome segregation movement was organized in a common spindle, and the daughter nuclei at the telophase appeared to envelope each other in the newly formed nuclear membrane. The cell was divided into two daughter cells by a common cell plate. Trinucleates were similarly converted to two cells with a hexaploid number of chromosomes. Some of the di- and trinucleates subsequently completed the second meiotic division with the formation of typical tetrad configurations. In giant cells with more than several nuclei, chromosomes separated at random but reaggregated into one giant resting nucleus, with no later cytokinesis. The rate of meiotic development in multinucleates was relatively slower in cells which contained greater numbers of nuclei.     

Liver chromatin fractions inMus andAkodon

Summary The liver chromatin fromMus musculus andAkodon molinae was separated in 8 fractions by differential centrifugation. Like fractions from both species showed approximately similar contents of DNA, equivalent ratios of histone to non-histone proteins, corresponding template activities and equal amounts of positive C-banded material. On the other hand, heavy chromatin fractions ofMus were highly enriched in satellite DNA whereas no satellite DNA was found inAkodon chromatin. Heavy chromatin fractions isolated by differential sedimentation have been usually homologued with the constitutive heterochromatin. The properties of the constitutive chromatin are discussed and the validity of the foregoing concept is challenged. It is proposed to define the constitutive heterochromatin as those chromatin regions comprising highly repeated DNA sequences clustered in restricted areas of chromosomes and not transcribed (satellite DNA).     

Endopolyploidy and structures of the nuclei in the endosperm of theCucurbitaceae

Summary In the endosperm ofEchinocystis lobata the endopolyploid nuclei show structural changes of chromocentres typical of the process of endomitosis in Angiosperms.The cytological differentiation of the endosperm proper and the chalazal haustorium in three further representatives of theCucurbitaceae: Sicyos angulata, Cyclanthera pedata andCitrullus colocynthis is also connected with endomitotic polyploidisation. The nuclei of the endosperm proper attain in the different species different levels of polyploidy, 384n, as a maximum. In the nuclei of the chalazal haustorium the degree of polyploidy is lower (as a maximum 24n).The structure of polyploid nuclei which did not undergo postendomitotic mitoses shows some differences in the species studied in the course of the present work.Sicyos angulata has prochromosomic diploid nuclei, whereas those ofCitrullus colocynthis andCyclanthera pedata can be assigned to chromomeric nuclei with a dense structure. Endopolyploid nuclei of all three species studied are characterized by the occurence of compact chromocentres of different size; besides in some nuclei ofSicyos angulata andCyclanthera pedata the chromocentres within the resting nucleus reveal their chromomere structure. In addition, inCitrullus colocynthis nuclei with numerous thin threads of various degrees of spiralisation have been observed. — In all three species the endopolyploid nuclei after mitotic divisions have notably higher numbers of nucleoli and small chromocentres.Dedicated to Professor L.Geitler on the occasion of his 70th birthday.     

Single fertilization inSpiranthes sinensis

The reproduction system inSpiranthes sinensis (Orchidaceae), collected at various localities in Japan, was revealed to be of a peculiar new type which is not to be found in other angiosperms. None of generative cell nuclei has been observed to dividede novo throughout the progamic phase, although they do participate in fertilization. Neither fertilization of the generative cell nucleus with the central nucleus nor formation of endosperm occurred in this plant. Although the chromosome number in the developing proembryonic cells numbered 2n=30, exhibiting diploidy, the number of marker chromosomes was equal to the sum of half the number of marker chromosomes of the parental clones. From the results described above, we may conclude that the reproduction system of this species represents a new type of single fertilization (non-double fertilization) between egg cell and sperm cell nuclei caused by the omission of generative cell division and the formation of only one sperm cell nucleus.     

Gene amplification in oocytes with 8 germinal vesicles from the tailed frog Ascaphus truei Stejneger

In the last 3 oogonial mitoses in Ascaphus truei all daughter nuclei remain in the same cell. The oocyte is 8-nucleate at the start of meiotic prophase and remains so until late in oogenesis when 7 of the nuclei disappear. All 8 nuclei in a single oocyte resemble one another with respect to size and chromatin distribution at all stages of meiotic prophase. Much of the Feulgen-positive material in pachytene nuclei is concentrated into one region of the nucleus. — All of the 8 germinal vesicles of yolky oocytes have a full set of lampbrush diplotene bivalents. Germinal vesicles from oocytes of up to 0.8 mm diameter have less than 100 nucleoli, some of which are multiple nucleoli in the sense that they have more than one core region. Each of the 8 nuclei in oocytes from one animal had about the same volume of nucleolar material. — Two values have been obtained for the amount of DNA in a diploid nucleus from Ascaphus. A biochemical estimate utilizing erythrocyte nuclei and the diphenylamine reaction yielded a value of 7.1 pg per nucleus. Microphotometry of erythrocyte nuclei stained with Feulgen's reagent gave a value of 8.2 pg per nucleus. — Microphotometric measurements of Feulgen-stained nuclei at various stages of meiotic prophase up to diplotene indicate that each nucleus synthesizes up to 5 pg of extrachromosomal DNA during and immediately after pachytene. This DNA is considered to be nucleolar. Autoradiography of nuclei from oocytes which had been incubated for 6h in ³H thymidine showed silver grains over pachytene and early diplotene nuclei only. In pachytene nuclei the silver grains overlaid that part of the nucleus where Feulgen-positive material was most concentrated. Most of the chromosomal material was unlabelled. — The significance of the 8-nucleate condition in Ascaphus oocytes is discussed, and the amount of nucleolar DNA synthesized at pachytene and of nucleolar material present in germinal vesicles is compared with corresponding situations in other amphibians.     

The ultrastructure of mitosis inIsochrysis galbana parke (Prymnesiophyceae)

Summary Mitosis and cytokinesis have been studied in the flagellate algaIsochrysis galbana Parke (Prymnesiophyceae). Nuclear division is preceded by replication of the flagella and haptonema, the Golgi body and the chloroplast; fission in the chloroplast occurs in the region of the pyrenoid. During prophase, spindle microtubules radiating from two ill-defined poles are formed. The nuclear envelope breaks down and the chromatin condenses. At metaphase the spindle is fully developed, some pole-to-pole microtubules passing through the well-defined chromatin plate, others terminating at it. No kinetochores or individual chromosomes were observed. By late metaphase, many Golgi-derived vesicles may be seen against the two poleward faces of the metaphase plate. During anaphase, the two daughter masses of chromatin move towards the poles. In early telophase, the nuclear envelope of each daughter nucleus is complete only on the side towards the adjacent chloroplast, remaining open on the interzonal side. However, during telophase each nucleus becomes reorientated so that it lies lateral to the long axis of the spindle and with its open side towards the chloroplasts. By late telophase, each new nuclear envelope is complete and confluence with the adjacent chloroplast ER established.Cytokinesis and subsequent segregation of the daughter cells are effected by the dilation of Golgi- and ER-derived vesicles in the interzonal region. No microtubular structures are involved. Comparisons with the results from other studies of mitosis in members of thePrymnesiophyceae show that they all have a number of features in common, but that there are differences in detail between species.     

Comparisons of nutrient recovery and specific leaf area variation betweenCarex lasiocarpa var.occultans andCarex thunbergii var.appendiculata with reference to nutrient conditions and shading byPhragmites australis

The distribution of two sedge species was studied in two mires which differ in abiotic environments and in distribution ofPhragmites australis. Carex lasiocarpa var.occultans dominated in nutrient-poor valley mire, andCarex thunbergii var.appendiculata dominated in nutrient-rich flood plain subject to water fluctuations.Phragmites australis grew well in nutrient-rich conditions. The distribution ofC. lasiocarpa showed a strong negative correlation withP. australis coverage, whereasC. thunbergii coverage was not affected byP. australis. The leaf area per dry leaf mass (specific leaf area: SLA) ofC. thunbergii increased with shading byP. australis, but that ofC. lasiocarpa was stable. The SLA flexibility ofC. thunbergii to light interception might enable this species to invadeP. australis patches in nutrient-rich environments. The residual nutrient ratio of nitrogen and phosphorus (the ratio of the residual nutrient content at the end of the growing season to peak nutrient content) in the vegetative ramet ofC. thunbergii was 1.7 times higher than that ofC. lasiocarpa. This low residual ratio may indicate effective nutrient recovery to storage organs. The effective nutrient recovery inC. lasiocarpa might enable this species to grow even in nutrient-poor environments. However, it may be difficult forC. lasiocarpa to expand its habitat to nutrient-rich areas whereP. australis dominates as it is not shade tolerant.     

Carbon input from plant to soil through root exudation inDigitaria adscendens andAmbrosia artemisiifolia

The amount of carbon released into soil through root exudation byDigitaria adscendens Herrm. andAmbrosia artemisiifolia L. var.elatior Desc., which often predominate at the early stages of secondary succession, was evaluated in a laboratory experiment conducted over a 60-day period. Differences in the amount of exuded carbon between these species and between the developmental stages were examined. The amount of carbon exuded increased with growth in both species. The percentage of exuded carbon to photosynthetically net fixed carbon, which was higher at younger stages (13%) inD. adscendens, decreased to 3.1% with time. On the other hand, no reduction in the amount of carbon exuded was observed inA. artemisiifolia (4.7–8.1% range). The total amount of carbon released through root exudation inD. adscendens andA. artemisiifolia was estimated at 3.1% and 6.9% of photosynthetically net fixed carbon, respectively. These results suggest the possibility that wild plants exude a considerable amount of carbon from their roots to the soil, and emphasizes the necessity for considering root exudation in the carbon cycle.     

Morphological changes of sperm nuclei during spermatogenesis in the brown alga Cystoseira hakodatensis (Fucales, Phaeophyceae)

Morphological changes and chromatin condensation of sperm nuclei were observed during spermatogenesis in the fucalean brown alga Cystoseira hakodatensis (Yendo) Fensholt. Ultrastructural studies have shown that the mature spermatozoid has an elongated and concave nucleus with condensed chromatin. The morphological changes and the chromatin condensation process during spermatogenesis was observed. Nuclear size decreased in two stages during spermatogenesis. During the first stage, spherical nuclei decreased in size as they were undergoing meiotic divisions and the subsequent mitoses within the antheridium. During the second stage, the morphological transformation from a spherical into an elongated nucleus occurred. Afterwards, chromatin condensed at the periphery in each nucleus, and chromatin‐free regions were observed in the center of the nucleus. These chromatin‐free regions in the center of nucleus were compressed by the peripheral chromatin‐condensed region. As the result, the elongated and concave nucleus of the mature sperm consisted of uniformly well‐condensed chromatin.     

Flavanol binding of nuclei from tree species

Light microscopy was used to examine the nuclei of five tree species with respect to the presence of flavanols. Flavanols develop a blue colouration in the presence of a special p-dimethylaminocinnamaldehyde (DMACA) reagent that enables those nuclei loaded with flavanols to be recognized. Staining of the nuclei was most pronounced in both Tsuga canadensis and Taxus baccata, variable in Metasequoia glyptostroboides, faint in Coffea arabica and minimal in Prunus avium. HPLC analysis showed that the five species contained substantial amounts of different flavanols such as catechin, epicatechin and proanthocyanidins. Quantitatively, total flavanols were quite different among the species. The nuclei themselves, as studied in Tsuga seed wings, were found to contain mainly catechin, much lower amounts of epicatechin and traces of proanthocyanidins. Blue-coloured nuclei located centrally in small cells were often found to maximally occupy up to 90% of a cells radius, and the surrounding small rim of cytoplasm was visibly free of flavanols. A survey of 34 gymnosperm and angiosperm species indicated that the first group has much higher nuclear binding capacities for flavanols than the second group.Abbreviations DMACA p-Dimethylaminocinnamaldehyde Communicated by W. Barz