ULTRASTRUCTURE OF MEIOSIS IN DASYA BAILLOUVIANA (RHODOPHYTA). I. PROPHASE I1

This first of two papers on ultrastructural observations of meiosis in the red alga Dasya baillouviana (Gmelin) Montague describes stages of prophase I of meiosis. Although the five stages of prophase were originally derived from light microscopic studies, the same stages were utilized for this study based on the developmental sequence of the synaptonemal complex, which has the same morphology and mode of development as those reported for other red algae. The cytoplasm in early prophase sporocytes was typically less electron dense than either vegetative cells or sporocytes in later stages of meiosis. The reduction in density suggests clearing of ribosomes and other cytoplasmic components prior to conversion from sporophyte to gametophyte control. Leptotene cells often had an amorphous, chromatin-free area, function unknown, which was not obviously associated with any specific nuclear region. Diplotene cells were characterized by nuclei containing prominent ring-shaped nucleoli composed of a dark staining ring of material surrounding an electron-translucent “vacuole.” Packets of electron-dense, fibrillar material were often noted in the cytoplasm of late prophase cells. These packets are thought to he “nuage,” a term applied to large cytoplasmic aggregations of RNA in germ cells of several other phyla. It is suggested that nuage may represent a new infusion of ribosomal and messenger RNA for post-meiotic development. The division pales are established by late prophase and a single polar ring is found within each large “exclusion zone” in close association with a pore-free area of nuclear envelope. Both annulate lamellae and small, numerous vesicles are located in the exclusion zones. The significance of the various aspects of prophase I is discussed with the overall observation that this phase of meiosis in red algae is very similar to the process in higher plant and animal cells.     

GROWTH AND DIFFERENTIATION OF AXIAL AND LATERAL FILAMENTS IN BATRACHOSPERMUM SIRODOTII (RHODOPHYTA)1

Development of the vegetative gametophyte of Batrachospermum sirodotii Skuja was examined with light and both transmission and scanning electron microscopy. Patterns of wall growth were followed using the Calcofluor White ST pulse-chase method. Thallus structure was analysed in terms of the pattern of development of the apical, periaxial and pleuridial initials that generate the axial and whorled lateral filaments characteristic of Batrachospermum. Apical cells of axial filaments elongate initially by tip growth with the nucleus maintaining a distal position. Nuclear division is horizontal. One daughter nucleus migrates basipetally and a thin, convoluted annular septum and perforate-occluded pit connection are then formed. Elongating axial cells subsequently extend by wall deposition at the base of the cell. Periaxial cells are initiated laterally and elongate primarily by tip growth while the nucleus remains within the axial cell. The nucleus then migrates to the boundary between the initial and the axial cell, divides, and one daughter nucleus moves into the initial and the other back into the axial cell. A slightly irregular annular septum and simple-occluded pit connection are then formed. Pleuridial cell initials begin as terminal to subterminal protuberances on periaxial or pleuridial cells. They first extend by tip growth and later by bipolar band growth. The nucleus remains within the parent cell as the pleuridial initial expands and a narrow septal ring is formed between the two cells. It then migrates through the septal ring into the initial and divides transversely. One nucleus passes back into the parent cell and a thick, flat septum and perforate-occluded pit connection are formed. It is concluded that the potentially indeterminate axial filaments and the determinate lateral pleuridia represent distinct developmental types in Batrachospermum.     

PHYCOBILIPROTEIN COMPOSITION AND CHLOROPLAST STRUCTURE IN THE FRESHWATER RED ALGA COMPSOPOGON COERULEUS (RHODOPHYTA)1

Serial sections of uncorticated axial cells of Compsopogon coeruleus revealed a single interconnected parietal chloroplast. Phycobilisomes in such chloroplasts were hemidiscoidal in shape with a broad-face diameter of ca. 25–30 nm. The molar ratio of phycobiliproteins in whole cell extracts was IPE:3PC:1APC, similar to isolated phycobilisomes. Two spectrally distinct C-phycocyanin forms (A618 nm, F648 nm and A630 nm, F652 nm) were resolved in dissociated phycobilisomes along with B-phycoerythrin and allophycocyanin.     

人早幼粒白血病细胞(HL-60)的表面超微结构及其在诱导分化过程中的变化

应用扫描电镜技术,结合超薄切片和组织化学方法,观察分析了人早幼粒白血病细胞(HL-60)的表面形态结构。扫描电镜下,HL-60细胞最明显的表面形态特征是绝大多数(约90%)具有丝状伪足。丝状伪足呈丝状、指状或圆棒状,其平均长度为4—6μ,直径约为0.25—0.3μ。丝状伪足具有明显特征性分布,常以成簇或成束状态分布在细胞的周围或一侧。超薄切片和组织化学反应结果显示,丝状伪足的内部结构主要是微丝和糖原颗粒。大多数HL-60细胞具皱褶而极少数细胞显示微绒??毛。用维生素A酸处理HL-60细胞6天后,有些细胞的丝状伪足消失,其细胞表面变得不规则,出现大型钝形伪足。由于丝状伪足与大型钝形伪足的成分基本相同,以及钝形伪足的出现伴随丝状伪足的减少,故推测钝形伪足可能来自丝状伪足。文中讨论了HL-60细胞形态结构与生理功能的关系。     

ULTRASTRUCTURE OF MEIOSIS IN DASYA BAILLOUVIANA (RHODOPHYTA). II. PROMETAPHASE I—TELOPHASE II AND POST-DIVISION NUCLEAR BEHAVIOR1

This is the second of two papers which together are the first comprehensive ultrastructural report of meiosis in a red alga. Many details of the meiotic process in Dasya baillouviana (Gmelin) Montagne are the same as those reported previously for mitotic cells in ceramialian red algae, but several characteristics seem unique to meiotic cells. The nucleus and nucleolus of meiotic cells are larger than those of mitotic cells and large accumulations of smooth ER are often found at the division poles during meiosis 1. The function of the ER accumulations is unknown. Importantly, both interkinesis and a simultaneous division of two separate nuclei during meiosis II was demonstrated. These new observations fail to support earlier speculation on higher red algae for a “uninuclear” meiosis (both nuclear divisions within the same nuclear envelope). However, following meiosis II the four nuclei migrate centripetally and possibly fuse in the center of the tetrasporangium. This post-division nuclear maneuvering is not understood, but our interpretation accounts for the earlier and erroneous impression of “uninuclear” meiosis. Perhaps the most important aspect of meiosis observed in Dasya is its basic adherence to the pattern commonly seen in higher plants and animals. This conservatism of the meiotic process lends further skepticism to the belief that red algae are extremely “primitive” organisms, although they undoubtedly represent a very “ancient” group of eukaryotic plants.     

ULTRASTRUCTURE OF EARLY DEVELOPMENT IN THE FEMALE REPRODUCTIVE SYSTEM OF POLYSIPHONIA HARVEYI BAILEY (CERAMIALES,RHODOPHYTA)1

Prefertilization and immediate postfertilization development in the female reproductive branch of Polysiphonia harveyi (Bailey) was studied with the electron microscope. Results pertaining to prefertilization morphology and development are consistent with those established in earlier light microscopic studies, but several unexpected ultrastructural characteristics were discovered. The mature carpogonium was found to have double membrane-bound vacuoles of nuclear origin and the carpogonial nucleus contained a nucleolus with a distinctive crystalline lattice. Trichogynes lacked a nucleus. Of particular interest was the discovery of a highly structured channel of smooth endoplasmic reticulum (SER) which extended uninterrupted, except for pit connections, through the carpogonial branch. It is suggested that the message of fertilization is conducted through the SER channel from the carpogonium to the support cell. Few observations were made on postfertilization branches, but evidence of direct fusion between the carpogonium and auxiliary cell was fairly conclusive. Pit plugs in the female branch were found to be of three morphological types differing by the presence and number of pit membranes. We have designated these plugs Type I–III since differential functioning was not fully ascertained. Our data suggest that pit membranes may serve to stabilize the plug and that those plugs without membranes are more readily dismantled to allow cytoplasmic continuity between cells.     

ULTRASTRUCTURE AND CYTOCHEMISTRY OF EARLY SPERMATANGIAL DEVELOPMENT IN ANTITHAMNION NIPPONICUM (CERAMIACEAE,RHODOPHYTA)1

Spermatial development and differentiation of wall components were investigated by electron microscopy and cytochemical methods in Antithamnion nipponicum Yamada et Inagaki. The spermatium is composed of two parts, a globular head and two appendages projecting from near the basal portion. The appendages originate form spermatangial vesicles (SVs) and follow a developmental sequence beginning as amorphous material and ending as fully formed fibrous structures compressed with in the SVs. SV formation is due to contributions initially from endoplasmic reticulum and later form dictyosome-derived vesicles. Chemical differentiation of the spermatial wall occurs early in its development. Calcofluor white ST does not label spermatial walls, indicating an absence of cellulose polysaccharides, which are abundant in vegetative cell walls. Labeled lectins show that α-d -methyl manose and / or α-d -glucose as well as N-acetyl-glucosamine, β-d -galactose, and α-l -fucose moieties are present on the spermatial wall but not in the vegetative cell wall. The glyconjugate with α-d -methyl mannose and / or glucose residues, previously reported as a gamete recognition molecule in this species, is distributed along the surface of spermatia as well as in the SV during spermatangial development.     

ULTRASTRUCTURE,DEVELOPMENT AND CYTOPLASMIC ROTATION OF SETA-BEARING CELLS OF COLEOCHAETE SCUTATA (CHLOROPHYCEAE)1

Part of the cytoplasm, which always contains the plastid, of seta-bearing cells of Coleochaete scutata Bréb. rotates clockwise about the base of the seta. Many golgi bodies, vesicles and much endoplasmic reticulum occupy the bridges between the rotating central core of cytoplasm and the stationary peripheral layer of these cells. The setae, which grow from their base, are devoid of organelles other than vesicles and elongate mitochondria. At irregular intervals along the thin seta wall are annular thickenings containing callose. Microtubules which encircle the base of the seta disappear on treatment with colchicine. This drug had no effect on the speed of rotational streaming or the growth rate of existing hairs but did inhibit the development of new setae. Cytochalasin B slowed, but did not stop, streaming after 3 h exposure. However caffeine, but not EDTA, EGTA or the Ca ionophore A23187, reversibly inhibited cyclosis. The mechanism of cytoplasmic rotation is discussed in the light of these drug treatments and the presence of actin in the alga.     

ULTRASTRUCTURE OF THE ZOOSPORES AND VEGETATIVE CELLS OF TETRAEDRON AND CHLOROTETRAEDRON (CHLOROPHYCEAE)1

The flagellar apparatus of the zoospores of Tetraedron bitridens Beck-Mannagetta and Chlorotetraedron polymorphum MacEntee, Bold et Archibald includes directly opposed basal bodies, a distal fiber that is elaborated into a ribbed structure to which the continuous striated microtubule-associated component (SMAC) is connected, and partial caps over the proximal end of each basal body. The angle between basal bodies ranges from approximately 25° to 150°. Basal bodies at wider angles are interconnected via their cores. A septum is present in the B-tubule of one basal body triplet in C. polymorphum. Both organisms have four microtubular rootlets arranged in a cruciate pattern. The two X-membered rootlets in a single cell have dissimilar numbers of microtubules. In C. polymorphum there are 5 and 6 microtubules in a 4/1 and 5/1 arrangement. 3/1 and 4/1 rootlets are present in T. bitridens. Zoospores of T. bitridens have a fuzzy coat whereas those of C. polymorphum are naked. Pyrenoids in both species are covered by a continuous starch sheath. Vegetative, interphase cells of C. polymorphum have two centrioles connected by a fiber that are located in depressions in the nuclear envelope. We propose that these two genera may be closely related to Neochloris, and that the coenobial genera Hydrodictyon, Pediastrum and Sorastrum are derived from a Tetraedron-like alga.     

ULTRASTRUCTURE AND CYTOCHEMISTRY OF SPERMATANGIAL DEVELOPMENT AND FERTILIZATION IN AGLAOTHAMNION OOSUMIENSE (CERAMIACEAE, RHODOPHYTA)

Freshwater dinoflagellates have not previously been reported from Belize, although there has been extensive work with marine dinoflagellates and some work with other freshwater groups. Freshwater dinoflagellates are more frequently observed in standing water and none have been observed in the several streams and rivers sampled since 1990. The goal in 1998 was to examine water samples from small ponds within hours of collection to improve the chance of observing swimming dinoflagellates. A plankton net was used and whole water samples also were collected. A small brown water pond on a peninsula and 30m from the Caribbean yielded a bloom of Thompsodinium intermedium. Dinoflagellates, including Peridinium centenniale , Katodinium sp., and Peridinium sp. in the Umbonatum Group, were observed within Crocodile pond and Lily pond on the mainland.     

THE ULTRASTRUCTURE OF MEIOSPORES OF COLEOCHAETE PULVINATA (CHAROPHYCEAE)1

In view of the relative importance of reproductive cell ultrastructure in phylogenetic and systematic studies of green algae, we investigated the fine structure of germinating zygotes and meiospores of Coleochaete pulvinata Braun. Meiospores have a flagellar apparatus very similar to that of zoospores and spermatozoids of the same species. Meiospores differ from zoospores and spermatozoids of C. pulvinata in having pyramidal body scales similar to those present on zoospores of C. scutata. Meiospores of C. pulvinata had as many as twice the number of spline microtubules as zoospores, and four times the number present in splines of spermatozoids of the same species. Developing meiospores of C.pulvinata, like those of other Coleochaete species, are individually surrounded by chamber walls. These differed from vegetative cell walls in lacking plasmodesmata. Moreover, the chamber walls in germinating zygotes of C.pulvinata stained a cobalt blue color with resorcinal blue, and fluoresced yellow in the presence of aniline blue, thus exhibiting the staining characteristics of callose. In location, morphology and presence of callose, chamberwalls resemble “special walls” of land plants, they may represent a charophycean spore development preadaptation useful in the evolution of walled spores characteristic of land     

ULTRASTRUCTURE OF TETRASPOROGENESIS IN PALMARIA PALMATA (RHODOPHYTA)1

The tetrasporangial initial in Palmaria palmata (L.) O. Kuntze (formerly Rhodymenia palmata (L.) Greville) arises from a cortex cell which enlarges and deposits a protein-rich wall layer. This cell undergoes mitosis to form a tetrasporocyte and a stalk cell. Synaptonemal complexes are formed in the sporocyte nucleus while in the cytoplasm floridean starch is deposited in association with ER or with particles presumed to be ribosomes. Microbody-like structures become numerous between the nuclear envelope and perinuclear ER, and clusters of non-membranous, spherical structures also are associated with the nucleus. Chromatin condensation is reversed following pachytene and a prolonged diffuse stage ensues, when dictyosomes and ER produce vesicles which deposit mucilage rich in sulfated and acidic polysaccharides around the tetrasporocyte. A conspicuous lenticular thickening of the mucilage sheath develops at the apical end of the sporangium. Dictyosomes are frequently associated with mitochondria which may be associated with chloroplasts. Following nuclear divisions the tetrasporocyte is cleaved into four spores by sequentially initiated, but simultaneously completed periclinal and anticlinal furrows. When mucilage deposition ceases, the dictyosomes begin to produce vesicles with glycoprotein-rich contents. These vesicles are abundant in released tetraspores, and they probably contain adhesive material aiding in the attachment of the liberated spores.     

ESTABLISHMENT,PERSISTENCE AND DOMINANCE OF CORALLINA (RHODOPHYTA) IN ALGAL TURF1

The dominance of Corallina thalli in an intertidal algal assemblage was examined by a series of algal removal experiments to test the hypothesis that other tam are competitively excluded from rock substrate. An alternate explanation, that environmental factors seasonally filter out taxa leaving Corallina as the only alga adapted for year to year survival, was considered. Development of vegetation on patches of naturally exposed bare rock was monitored and compared with manipulated surfaces. Thalli of several species were selectively removed from exposed surfaces and intact turf;. changes in tam occupying primary substrate were recorded over more than three years. No significant differences in mean percent cover for Corallina, Lithothrix, bare rock, or algal crusts were found among treatments. Except for initial growth of colonizing species, abundances of other species dad not increase in the absence of Corallina. Large amounts of rock remained bare or intermittently covered by transient populations of short-lived algae. Data from single quad-rats, where individual clumps of Lithothrix were followed from month to month, indicated that the continued presence of this co-occurring and often abundant species depended on turnover of short-lived thalli rather than persistence of the same clumps. No interactions were found among the several categories of species that appeared after rock substrate was cleared. Most species were the same ones that grew, epiphytically in intact turf at the same time. In control quadrats Corallina maintained 59-95% cover while slowly increasing on surfaces earlier exposed. I predict that Corallina species will regain their dominance in the absence of competitors for primary substrate if the slowly spreading basal crusts remain undisturbed. Morphological and life history characteristics are identified that adapt Corallina to its dominant role in this habitat.     

A STARCH GRAIN-MITOCHONDRION-DICTYOSOME ASSOCIATION IN BATRACHOSPERMUM(RHODOPHYTA)1

In Batrachospermum sirodotii Skuja the dictyosome-mitochondrion association, commonly reported in other red algae, includes a floridean starch grain during periods of intense dictyosomal activity. Such an organellar assemblage may serve to provide the most efficient means of energy flow during periods of peak dictyosomal activity.     

ULTRASTRUCTURE OF GERMINATING CARPOSPORES OF PORPHYRA VARIEGATA (KJELLM.) HUS (BANGIALES,RHODOPHYTA)1

The fine structure of released, attached, and germinating carpospores of Porphyra variegata (Kjellm.) Hus is described. Adhesive vesicles, formed during sporogenesis and discharged upon settling of the spore, produced a layer of adhesive mucilage around the spore and filled a deep imagination on the spore's ventral side. The mucilage layer was punctured by the emergence of a germ tube. Both spore and germ tube were lined by newly deposited cell wall. Germination was accompanied by vacuolation and starch mobilization. The morphological development of the sporeling was not noticeably influenced by the great variability of the timing, location, and orientation of septum formation. The attached carpospore possessed a plastid like that of gametophyte cells: stellate with one large central pyrenoid and no peripheral encircling thylakoids. Cells of mature vegetative cells of the conchocelis had plastids that were elongate and parietal and had multiple pyrenoids and encircling thylakoids. Most stages in the transition between the two forms of plastids occurred during carpospore germination.     

THALLUS MORPHOLOGY AND SPORE FORMATION IN CULTURED ERYTHROCLADIA IRREGULARIS ROSENVINGE (RHODOPHYTA,BANGIOPHYCIDAE)1

The discoid thallus of Erythrocladia irregularis is the result of a pattern of cellular divisions expressed by mono-and endospores; the latter are described for the first time in Erythrocladia. In nutrient enriched growth conditions the discoid thallus becomes a shapeless aggregate of unicells with a wrinkled wall surface, whose plane of division is unpredictable. The cells are able to produce monosporangia with a curved wall or they form eight-celled endosporangia. Chloroplasts have an internal pyrenoid, as is characteristic of the Erythropeltidaceae. The observation that E. irregularis can form either a colonial cluster of unicells or a discoid thallus would lead to the assignment of the genus to the Porphyridiales; however, since this order is reserved to organisms in which only the unicellular organization can be expressed, it is suggested to maintain Erythrocladia in the Erythropeltidales.     

ATTACHMENT AND FUSION OF GAMETES DURING FERTILIZATION OF PALMARIA SP. (RHODOPHYTA)1

Fertilization of cultured microscopic female gametophytes by spermatia from field-collected male gametophytes of Palmaria sp. was observed by light and transmission electron microscopy. Liberated spermatia had a prophase-arrested nucleus with a pair of polar rings. The protoplast of spermatia was covered with ca. a 3-μm-thick hyaline covering. After spermatium inoculation, the spermatial covering was attached specifically to the coat surrounding the cell wall of the trichogyne. The spermatial covering was eliminated only at the site of gamete attachment, resulting in direct attachment of the spermatial plasma membrane to the trichogyne within 5 min after spermatium inoculation. This direct attachment was followed by completion of spermatial nuclear division and cell wall formation. The polar rings disappeared before prometaphase. The cytoplasm of the binucleate spermatium invaded the trichogyne cell wall and subsequently fused with the trichogyne cytoplasm. The trichogyne could fuse with many spermatia, and many male nuclei (the derivative nuclei of spermatial nuclear division) could enter the trichogyne cytoplasm.     

AN HERBICIDE (SULFOMETURON METHYL) RESISTANT MUTANT IN PORPHYRIDIUM (RHODOPHYTA)1

The unicellular red alga Porphyridium sp. is, like many bacteria, fungi and higher plants, sensitive to the sulfonylurea herbicide sulfometuron methyl (SMM). However, the minimal inhibitory concentration for SMM in Porphyridium (55 μM) is higher than in green plants. We isolated a spontaneous SMM-resistant mutant SMR1 of Porphyridium sp. by use of a double-layered agar technique. The mutation frequency and the similarity of the mutant's morphology and growth patterns to the parent strain strongly suggest that SMR1 is a single gene mutation. The activity of the enzyme acetohydroxy acid synthase (AHAS) in crude extracts of the mutant SMR1 is at least two orders of magnitude less sensitive to SMM than that of the parent strain, which indicates that AHAS is the target of SMM (as has been shown in a variety of organisms). We propose that such a mutation, the first isolated in a unicellular rhodophyte, would be a useful marker for genetic studies of Porphyridium. It may also be useful for maintaining unialgal cultures in large scale open ponds.     

植物分类学在化石珊瑚藻(珊瑚藻目,红藻门)中的应用

最近有人认为将化石藻类的分类归入现生藻类分类单元有利于珊瑚藻作为古环境的标志,便于理解该类群的演化。然而,这样分类可能很难,因为并不是所有现生藻类分类特征都能在化石种中保存下来。Sporolithacea科的钙化部分(独立或者聚集的孢子囊群)的出现,可以把它们与这个类群的另一个现生科Corallinaceae区别开,这个科在生殖窠中产生孢子囊。节片的有无,丝间细胞的联系类型,生殖窠中孢子囊释放的数目都是用来划分Coral1inaceae科的亚科的标准,在化石样品中也可以用合适的条件进行观察。在大多数情况下,对现生珊瑚藻类属的划分特征可以在化石藻类中鉴别出来,但在几种现生珊瑚藻没有钙化的生殖结构或发育特征。因此,它们生殖结构无法与相应的化石藻类进行对比,也不能进行化石藻类的分类。近年来的趋势认为生殖结构和发育特征是对现生珊瑚藻进行分类的优先鉴定标准,然而,某些特征的稳定性在属的划分上仍然存在争论。在许多情况下,现生藻类的分类标准特征都不能在化石中保存,对古生物化石的分类标准的最佳选择是在化石藻类中选择辅助的,并且可以识别的其它鉴定特征,或者应用非正式的比现生藻类代表定义更宽的属名。