Two newhalophytophthora species,H. tartarea andH. masteri,from intertidal decomposing leaves in saltmarsh and mangrove regions

Two new pythiaceous fungi were obtained from decaying leaves submerged in saltmarsh environments of the east coast of the U.S.A., or fringing mangroves in the Bahama Islands, and are described here asHalophytophthora tartarea andH. masteri. The two species have superficially similar zoosporangia whose dehiscence tubes have ragged-looking apices. However, differences in fine structures and development of the dehiscence tube and plug, characteristics of the dehiscence plugs, and presence or absence of vesicles clearly distinguish the two species.Halophytophthora masteri is the only species ofHalophytophthora that has a zoospore release mechanism involving both an extruded plug and a vesicle. Cultural properties concerning growth and asexual reproduction at various salinities and temperatures are also different between the two, probably reflecting adaptation to their respective habitats. Though zoospore release inH. masteri occurs spontaneously from mature zoosporangia, it is remarkably enhanced inH. tartarea and alsoH. masteri by mildly dehydrating mature zoosporangia followed by rewetting with seawater, which suggests a possible relation between the asexual reproduction of these oomycetes and the tidal rhythm in their natural habitats.     

ALKALINE EARTH ELEMENTS AND ZOOSPORE RELEASE AND DEVELOPMENT IN PROTOSIPHON BOTRYOIDES

O'Kelley , Joseph C., and Walter R. Herndon . (U. Alabama, University.) Alkaline earth elements and zoospore release and development in Protosiphon botryoides . Amer. Jour. Bot. 48(9): 796–802. Illus. 1961.—Cells of Protosiphon botryoides Klebs from depleted nutrient medium containing Ca were washed and resuspended in fresh complete medium with Ca; or in media with a Sr, Ba or Na replacement, respectively, for Ca; or in an equivalent CaCl₂ solution or deionized water. Zoospore release was observed in these media upon illumination following a 12-hr dark period. Free zoospores were less abundant in Sr-, Ba- and Na-replacement media than in the Ca medium. Zoospore production and release also were depressed in solutions of only CaCl₂ and in deionized water. In the Sr and Ba media, zoospores were formed but not released from the parent cell, as a rule; some zoospores were released in mass within a gelatinous vesicle which did not liquefy and set the zoospores free; these zoospores lost motility and continued development in Sr, producing characteristic, spheroidal clusters of aplanospores. In the Na medium, protoplasmic cleavage preceding zoospore formation was severely inhibited. A study of the reversibility of Sr inhibition of the zoospore-release mechanism revealed evidence of reversion 12 hr after replacement of Sr by Ca. Walls of cells produced in Ca are rich in ruthenium red-positive materials, whereas cells produced under conditions of Sr replacement lack these materials. The significance of these findings in relation to the Ca requirement of other algal species is discussed.     

EFFECT OF NITROGEN,SULFUR AND OTHER FACTORS ON ZOOSPORE PRODUCTION BY PROTOSIPHON BOTRYOIDES

O'Kelley , J. C., and T. R. Deason . (U. Alabama, University.) Effect of nitrogen, sulfur and other factors on zoospore production by Protosiphon botryoides. Amer. Jour. Bot. 49(7): 771–777. Illus. 1962.—Nutrient-medium pH, osmotic pressure, staling products, and mineral depletion were studied in relation to zoospore production by Protosiphon botryoides in liquid media. In an intermediate range (pH 5.1–7.7), pH has little or no influence specifically on zoospore production. Although distilled water is an unsuitable medium for zoospore production, if a balanced nutrient medium is supplied, osmotic pressure is without pronounced influence over a wide range. Staling products in old cultures exert a minor inhibitory effect. Deficiencies of nitrate, sulfate, or calcium in liquid media can decrease drastically zoospore production or release, and nitrate or calcium depletion appears to be mainly responsible for loss of the capacity of Protosiphon to produce and release zoospores in aged liquid cultures.     

Survival and motility of diatoms<Emphasis Type="Italic">Navicula grimmei</Emphasis> and<Emphasis Type="Italic">Nitzschia palea</Emphasis> affected by some physical and chemical factors

Navicula grimmei and Nitzschia palea occurring almost equally in a mixed population on department moist garden soil surface when maintained in fresh supernatant (of soil-water medium) at pH 7.0, temperature of 26 +/- 1 degree C and under continuous light (intensity of approximately equal to 30 micromol m(-2) s(-1)) in a culture chamber exhibited a similar cell survival period (of 28 d) and percentage (at the beginning 100 % and mid of survival period 65 %) and stop gliding 11 d prior to cell death (with gliding speed reduced in both from 204-330 microm/min at the beginning to 82.5-99 microm/min at the mid of gliding period) irrespective of their size differences. However, a sharp fall in the cell gliding period, gliding cell percentage and speed occurred at various levels (different from cell survival period and percentage) in both diatoms in a similar extent under water stress (2, 4 and 6 % agarized supernatant, liquid supernatant with 0.2-1.0 mol/L NaCl, blot-dryness of cells for 5-15 min), pH extreme of liquid supernatant (< or =5.0, > or =9.0), temperature extremes in liquid supernatant (< or =15, > or =40 degrees C), UV exposure (0.96-5.76 kJ/m2), lack of all nutrients from the medium (double distilled water), darkness or low light intensities (2 and 10 micromol m(-2) s(-1)), presence of 'heavy' metals (Ni, Cu, Zn, Co, Fe, Hg; 1-200 ppm), organic substances in liquid supernatant (DDT, captan, urea, 2,4-D, 100-2000 ppm; thiourea, 50-1000 ppm). N. palea sway (turn around at either ends) or not only when gliding but independent of cell gliding speed, which decreased continuously under all conditions.     

In vitro production of zoospores by the mosquito pathogen Lagenidium giganteum (Oomycetes: Lagenidiales) on solid media

Reliable, large-scale production of Lagenidium giganteum zoospores was obtained on solid media. The fungus was grown for 7 days in a liquid medium of wheat germ, hemp seed, yeast extract, and glucose, then placed onto hemp-seed agar. Zoosporogenesis was induced on agar by immersing the fungal cultures into water. Zoospore production began 10 hr postimmersion, peaked at 18 hr, and ceased by 36 hr. A single, 10-cm Petri dish of fungus on hemp-seed agar produced 1.7?3.8 × 10⁷ zoospores during the 26 hr of zoosporogenesis. Optimal zoospore production occurred with 4- to 7-day-old cultures; cultures older than 10 days produced few zoospores. The temperature range for zoosporogenesis was 15–35°C. The extent of zoosporogenesis was directly related to the volume of water used to induce zoospore formation and inversely proportional to agar thickness. Bioassay of zoospores against second instar Culex quinquefasciatus larvae yielded an LD₅₀ of 400 zoospores/ml.     

Vegetative survival,akinete and zoosporangium formation and germination in some selected algae as affected by nutrients,pH, metals,and pesticides

Lack of nitrogen, phosphorus, or all nutrients, extremes of pH (<4, >11), presence of ‘heavy’ metals (Co, Cu, Zn, Hg, Pb; 0.5–10 ppm) or pesticides (carbofuran, 2,4-D, dithane, phorate, or bavistin; 1–50 ppm) decreased to various extent or completely inhibited the survival of vegetative cells in all studied algae. The formation of akinetes, thier viability and germination inAnabaena iyengarii, Westiellopsis prolifica, Nostochopsis lobatus andPithophora oedogonia and the formation of zoosporangia, their viability, and the germination of zoospores inCladophora glomerata andRhizoclonium hieroglyphicum was affected. The formation of viable akinetes or zoosporangia was shown to be directly linked with vegetative cell survival and growth; it could not be induced by any chemical stress imposed.     

The effect of the weight of the seedling-derived tuber on subsequent clonal generations in a potato breeding programme

Cultures of Polymyxa graminis were maintained in roots of barley plants grown in sand at different temperatures using Wisconsin soil temperature tanks. At 17 – 20°C, the minimum time from inoculation with cystosori to the production of zoospores from the inoculated roots was 2 – 3 wk. At 11 – 20°C many zoospores were produced but the incubation period was longer at the lower temperatures. Above 20°C little fungal development occurred. The duration of motility of zoospores ranged from c. 1 h to > 24 h. Bovine serum albumen (BSA) prolonged motility but glycine and glucose had no effect or, at higher concentrations, were toxic. Zoospores were rapidly immobilised by zinc ions in solution at or above 10μg/ml. In some experiments BSA added to the zoospore suspension greatly increased transmission of barley yellow mosaic virus (BaYMV) while glucose, glycine and ovalbumen decreased it. When seedlings were incubated with zoospore suspensions for 24 h at different temperatures, BaYMV transmission was high (> 60%) at 10, 15 and 20°C but there was little at 5 or 25°C. In experiments to determine the time taken for zoospore penetration, seedlings were incubated in suspension for different periods of time and then rinsed in zinc sulphate solution to kill free zoospores. Between 3 and 3·5 h was needed for zoospores to establish infection. Transmission occurred equally to plants of various ages between 3 days and 7·5 wk.     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture III. Some notes on the process of zoospore liberation

When Chlamydomonas reinhardi cells liberate zoospores, theyexcrete into the medium a factor(s) which induces zoospore liberationof other cells that are not yet ready to liberate zoosporesby themselves. The "factor" is contained within cells at laterstages of the cell cycle, but its action is suppressed untilthe regular time of zoospore liberation in the cell cycle. (Received November 18, 1974; )     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture I. Some characteristics of the cell cycle

Cells of Chlamydomonas reinhardi Dangeard were grown synchronouslyunder a 12 hr light-12 hr dark regime. Time courses of nucleardivision, chloroplast division, "apparent cytokinesis" and zoosporeliberation were followed during the vegetative cell cycle inthe synchronous culture. Liberation of zoospores occurred atabout 23–24 hr after the beginning of the light periodat 25°C. Four zoospores were produced per mother cell underthe conditions used. At lower temperatures, the process of zoosporeliberation as well as length of the cell cycle was markedlyprolonged, but the number of zoospores produced per mother cellwas approximately the same. At different light intensities,lengths of the cell cycle were virtually the same, while thenumber of zoospores liberated was larger at higher rather thanat lower light intensities. During the dark period, nuclear division, chloroplast divisionand apparent cytokinesis took place, in diis order, and proceededless synchronously than did the process of zoospore liberation.When the 12 hr dark period was replaced with a 12 hr light periodduring one cycle, the time of initiation as well as the durationof zoospore liberation was litde affected in most cases, whereasnuclear division, chloroplast division and apparent cytokinesiswere considerably accelerated by extended illumination. Whenalgal cells which had been exposed to light for 24 hr were furtherincubated in the light, zoospore liberation started much earlierand proceeded far less synchronously, compared with that under12 hr light-12 hr dark alternation. (Received October 12, 1970; )     

Establishment and maintenance of nuclear position during zoospore formation in allomyces macrogynus: roles of the cytoskeleton

The involvement of the microtubule (MT) and actin microfilament (MF) cytoskeletons in establishing nuclear positions during zoosporogenesis in Allomyces macrogynus was assessed using selective cytoskeletal disrupting treatments and documented with light microscopy. These experiments were coupled with low-speed centrifugation studies to determine the degree to which cytoskeletal elements anchor nuclear position. At the onset of zoospore formation, nuclei were positioned only in cortical cytoplasmic regions of the zoosporangia (ZS). Immunofluorescence microscopy revealed that MTs primarily emanated from centrosomal regions into the surrounding cytoplasm at this stage. During delimitation of the cytoplasm into individual uninucleate zoospores, nuclei migrated from cortical regions to become distributed throughout the cytoplasm. Coincident with nuclear migrations, MTs were primarily organized at and emanated from nuclear surfaces, forming extensive perinuclear arrays. Nuclear migrations were suppressed in ZS induced to sporulate in the presence of cytochalasin D, an actin MF inhibiting compound. Disruption of MTs with nocodazole did not block nuclear migrations, although resultant nuclear spacing was irregular. Centrifugation treatments of control and drug-treated ZS demonstrated that nuclear positions were stabilized by perinuclear MT arrays. The results indicate that nuclear motility in ZS of A. macrogynus is the result of an actin-based system while perinuclear MTs arrays function to establish and fix nuclear position during zoospore formation. Copyright 1998 Academic Press.     

Zoospore production and motility of mangrove thraustochytrids from Hong Kong under various salinities

We investigated the effects of salinity on the zoospore production of four mangrove thraustochytrid isolates, Schizochytrium sp. KF1, Aurantiochytrium mangrovei KF6, Thraustochytrium striatum KF9 and Ulkenia sp. KF13. The zoospore motilities, which were based on curvilinear velocity (VCL) and straight-line velocity (VSL), were monitored using the Computer-Assisted Sperm Motility Analysis (CASA) Software system. The zoospore production of four isolates was suppressed at salinity above 15‰. Schizochytrium sp. produced the greatest number of zoospores at 15‰, while Aurantiochytrium mangrovei and Ulkenia sp. produced abundant zoospores in diluted sea water ranging from 7.5 to 15‰. Thraustochytrium striatum performed relatively poorly under all salinities. Salinity and exposure time, as well as their interactions, had significant impacts on most zoospore velocity measurements. The optimal velocities of zoospore motility also varied among isolates. Zoospores of Schizochytrium sp. and A. mangrovei had similar responses to salinity, with the highest motility at 7.3‰, followed by a decrease in velocities with increasing salinity. In contrast, the zoospore of T. striatum had optimal motility at 12‰ and remained highly motile from 15 to 20‰. The velocities of zoospores of Ulkenia sp. were the lowest among the tested thraustochytrids and had optimal motility at 12‰. Zoospores of all the isolates remained active after 4 h of exposure to aqueous medium, but the optimal salinity for each mode of swimming changed. The ecological significance of these data are discussed.     

Regulation of attachment, germination, and appressorium formation by zoospores ofLagenidium giganteum and related oomycetes by chitin, chitosan, and catecholamines

Summary Lagenidium giganteum (Oomycetes: Lagenidiales), a facultative parasite of mosquito larvae, infects the larval stage of most species of mosquitoes and a very limited number of alternate hosts. Host infection by this and other members of Oomycetes is initiated by motile, laterally biflagellate zoospores. Chemical bases for the various degrees of host specificity exhibited by these parasites is not known, but presumably involves receptors on the zoospore surface recognizing compounds either secreted by or on the surface of their hosts. Surface topography had no detectable effect onL. giganteum encystment or appressorium formation. Scanning electron microscopy documented the detachment of flagella during zoospore encystment. Bulbous knobs at the basal end of the detached flagellum were interpreted as encysting zoospores dropping the axoneme and/or the basal body and associated structures to which flagella are attached. Multiple signals appear to be involved in the initial steps ofL. giganteum host invasion. Zoospores of this parasite did not encyst on powdered preparations of chitin or chitosan (deacetylated chitin). Upon dissolution of chitosan in dilute acid followed by drying these solutions to form thin, transparent films, zoospores readily encysted. The degree of reacetylation of these films and the spacing of acetylated and deacetylated residues had no significant effect on zoospore encystment. Zoospores of a strain ofLagenidium myophilum isolated from marine shrimp, that also infects mosquito larvae, encysted on chitosan films. No encystment of spores of the plant parasitePhytophthora capsici was observed on chitin or chitosan films. Simulation of cuticle sclerotization by incubating chitosan films with different catecholamines and tyrosinase significantly reduced zoospore encystment. Zoospores that encysted on chitosan films did not germinate in distilled water. Germination could be induced by adding microgram quantities of bovine serum albumin or proteins secreted by motile zoospores into the water, and to a lesser degree by some amino acids, but not by various cations. Zoospores encysted and germinated on the pupal stage of some mosquito species. Appressoria were occasionally formed, but most subsequently sent out another mycelial branch, apparently without attempting to pierce the pupal cuticle. Methylation of pupal exuviae with ethereal diazomethane or methanol/HCl significantly increased zoospore encystment. Modification of chitin by catecholamines, lipids and protein on the epicuticular larval surface all affected host invasion.Abbreviations BSA bovine serum albumin - CID collision-induced dissociation - DOPA 3,4-dihydroxyphenylalanine - ESI-MS electrospray mass spectrometry - ESI-MS/MS tandem electrospray mass spectrometry - SDS-PAGE sodium dodecyl sulphate polyacrylamide gel electrophoresis - WGA wheat germ agglutinin - ZAP zoospore aggregation pheromone     

A STUDY OF LIGHT INTENSITY,PERIODICITY, AND WAVELENGTH ON ZOOSPORE PRODUCTION BY PROTOSIPHON BOTRYOIDES KLEBS12

When mature Protosiphon cells were placed in darkness, zoospore production was more extensive and was completed in a shorter time at a temperature of 27 C than at 22 or 15 C. Cool-white fluorescent (Sylvania) light inhibited the process measurably at a radiation intensity of 0.6±10³ ergsjcm²-sec; inhibition was 96% complete at 14±10³ ergs/cm²-sec. For mature cells previously grown under repeated 12-12 hr light-dark cycles, a dark period of approximately 2 hr at 22 C allowed cell division to proceed to a stage such that reillumination did not inhibit continued development of zoospores. Monochromatic light from 402 to approximately -494 nm, as compared to darkness, inhibited zoospore formation; maximal inhibition was at 432-461 nm. In contrast, monochromatic light from 522 to 726 nm stimulated zoospore formation relative to darkness. Synchronous zoospore production was obtained using the following regimes: (A) 12 hr cool-white alternated with 12 hr yellow, (B) 12 hr cool-white alternated with 12 hr blue. Under regime A synchronous zoospore release (following synchronous production) occurred near the end of the yellow irradiation period, while under regime B it occurred near the end of the cool-white irradiation period. The significance of this in terms of photoprocesses and possible photoreceptors is discussed.     

Protein kinase C is likely to be involved in zoosporogenesis and maintenance of flagellar motility in the peronosporomycete zoospores

The motility of zoospores is critical in the disease cycles of Peronosporomycetes that cause devastating diseases in plants, fishes, vertebrates, and microbes. In the course of screening for secondary metabolites, we found that ethyl acetate extracts of a marine Streptomyces sp. strain B5136 rapidly impaired the motility of zoospores of the grapevine downy mildew pathogen Plasmopara viticola at 0.1 μg/ml. The active principle in the extracts was identified as staurosporine, a known broad-spectrum inhibitor of protein kinases, including protein kinase C (PKC). In the presence of staurosporine (2 nM), zoospores moved very slowly in their axis or spun in tight circles, instead of displaying straight swimming in a helical fashion. Compounds such as K-252a, K-252b, and K-252c structurally related to staurosporine also impaired the motility of zoospores in a similar manner but at varying doses. Among the 22 known kinase inhibitors tested, the PKC inhibitor chelerythrine was the most potent to arrest the motility of zoospores at concentrations starting from 5 nM. Inhibitors that targeted kinase pathways other than PKC pathways did not practically show any activity in impairing zoospore motility. Interestingly, both staurosporine (5 nM) and chelerythrine (10 nM) also inhibited the release of zoospores from the P. viticola sporangia in a dose-dependent manner. In addition, staurosporine completely suppressed downy mildew disease in grapevine leaves at 2 μM, suggesting the potential of small-molecule PKC inhibitors for the control of peronosporomycete phytopathogens. Taken together, these results suggest that PKC is likely to be a key signaling mediator associated with zoosporogenesis and the maintenance of flagellar motility in peronosporomycete zoospores.     

Factors influencing the sporulation and cyst formation of Aphanomyces invadans, etiological agent of ulcerative mycosis in Atlantic menhaden, Brevoortia tyrannus

Oomycete infections caused by Aphanomyces invadans occur in freshwater and estuarine fishes around the world. Along the east coast of the USA, skin ulcers caused by A. invadans are prevalent in Atlantic menhaden, Brevoortia tyrannus. From laboratory observations low salinities appear crucial to transmission of the pathogen. To better understand aspects of transmission, we characterized sporulation and cyst formation of secondary zoospores of two isolates of A. invadans at different salinities and temperatures. Sporulation occurred only at low salinities. At room temperature (ca. 20-22 C), using "pond water" augmented with artificial sea salts, the endemic strain WIC and the Thailand strain PA7 of A. invadans produced free-swimming secondary zoospores at salinities of 0, 1 and 2 psu (practical salinity unit = per thousand), but not at 4 psu or higher. Secondary zoospores of another species, ATCC-62427 (Aphanomyces sp.), were observed at 1, 2, 4 and 8 psu but not at 0 and 12 psu. Secondary zoospores of all three isolates, especially WIC, were abundant and motile 1-2 d postsporulation. Sporulation was temperature dependent and occurred over a relatively narrow range. No sporulation occurred at 4, 30 or 35 C for either WIC or PA7. For both strains zoospore production within 1-3 d after the initiation of sporulation was more prolific at 25 C than at 20 and 15 C. At 15 C production of zoospores was sustained over 11 d for WIC and 5 d for PA7. At room temperature single WIC secondary zoospores remained motile 12-18 h. Salinities exceeding 4 psu or vigorous shaking caused immediate cyst formation of WIC secondary zoospores. Exposure to menhaden tissue, but not tissues of other fishes to secondary zoospores (WIC), caused rapid (2 h) cyst formation. Cysts were capable of excysting when transferred to 1 psu water within 2-3 h of cyst formation. Cysts that had remained encysted in 6.5 psu for 24 h did not excyst when transferred to 1 psu water. Salinity and temperature requirements for sporulation indicate that juvenile menhaden must acquire infections during rain or in low salinity oligohaline waters.     

Pattern formation and the fine structure of the developing cell wall in colonies of Pediastrum boryanum

A single-layered disc of peripheral pronged cells and central prongless cells impart the typical gear shape to colonies of Pediastrum, while the walls of each cell have a characteristic reticulate triangular pattern. The two-layered wall forms in the cells during colony formation following zoospore aggregation and adhesion. The uniformly thin outer layer reflects contours resulting from differential thickening in the reticulate pattern of the inner, thicker, more fibrillar and granular wall layer. The reticulate pattern thus imparted to the outer wall layer persists in empty zoosporangia following the release of zoospores. Columns of electron-dense material extend through the outer wall layer except at the ridges and centers of the reticulum. Following mitosis and cleavage, the resulting zoospores are extruded within a vesicle membrane consisting of the inner wall layer. Separation of this membrane from the parent cell occurs in material of the inner layer adjacent to the outer wall. Vesicles containing swarming zoospores also contain a granular material which appears to become associated with the aggregating and adhering cells of new colonies. Microtubules occur in zoospores prior to adherence but are absent during wall deposition.     

Morphology of three polycentric rumen fungi and description of a procedure for the induction of zoosporogenesis and release of zoospores in cultures

Three polycentric rumen fungi, LL, LC2 and Ruminomyces elegans (C2), isolated from the rumen of cattle were grown in six culture media. LL and LC2 were morphologically similar. Their characteristics resembled those of Orpinomyces and Neocallimastix joyonii, and they grew well and produced sporangia after 3-4 d growth in all the media. R. elegans differed morphologically from LL and LC2, but although it also grew well in all media, abundant sporangia occurred only after 2-3 d growth in media containing cellulose. Undifferentiated sporangia were produced by all three isolates; differentiation of the sporangia did not occur in the spent growth media. However, if thalli possessing recently-formed sporangia were transferred to, or flooded with, fresh liquid medium or rumen fluid, zoosporogenesis and liberation of zoospores occurred within 17-20 min for isolates LL and LC2 and 30 min for R. elegans. Procedures for inducing zoosporogenesis by polycentric anaerobic fungi are described.     

Characterization of<Emphasis Type="Italic"> Aspergillus oryzae</Emphasis> fermentation extract effects on the rumen fungus<Emphasis Type="Italic"> Neocallimastix frontalis</Emphasis>, EB 188. Part 1. Zoospore development and physiology

Experiments were performed to determine the effect of Aspergillus oryzae (AO) fermentation extract on zoospore development in the rumen fungus Neocallimastix frontalis EB 188. Powdered product, or liquid extract prepared from such powder, was added at the recommended value for supplementation in dairy cattle. Stationary and stirred cultures were periodically sampled and assayed for extracellular and intracellular protein and enzymes, gas production, zoospore production and maturation, and carbon source utilization. Soluble extract increased fungal physiology when grown in stirred vessels or stationary cultures. Treated cultures produced higher levels of enzymes (nearly double). Mobile zoospores matured into germination entities more rapidly in treated cultures, and when powdered product was used, nearly 3 times more motile zoospores were produced at 56 h of fungal growth. Levels of the intracellular enzyme malate dehydrogenase increased by 6-fold in the presence of powdered product. Product wheat bran carrier used as soluble extract or powder had very little effect on fungal cultures. Medium cellulose was completely hydrolyzed in all cultures but this occurred earlier in those containing AO treatment.     

ULTRASTRUCTURE OF CEPHALEUROS VIRESCENS (CHROOLEPIDACEAE; CHLOROPHYTA). III. ZOOSPORES

Transmission electron microscopic examination of Cephaleuros virescens Kunze growing on leaves of Camellia spp. and Magnolia grandiflora L. indicates that unreleased zoospores in mature zoosporangia are similar to those produced by the related genus Phycopeltis epiphyton Millardet and unlike the quadriflagellate motile cells produced by taxa in other families of Chlorophyta. The zoospores bear four smooth isokont bilaterally “keeled” flagella containing typical “9 + 2” axonemes and lacking scales. Flagellar insertion is apical and the parallel basal bodies overlap laterally at two levels. A cross section through the four basal bodies shows a trapezoidal arrangement wherein the two upper (anterior) basal bodies are closer together than are the lower (posterior) two. Serial sections indicate that diagonally opposing upper and lower basal bodies anchor flagella which emerge from the same side of the apical papilla. Each of the four basal bodies is associated with a microtubular spline which extends beneath the plasmalemma to the posterior end of the zoospore. A distinct multilayered structure is associated with each of the lower basal bodies. A nucleus, mitochondria (two of which are closely associated with the nucleus and spline microtubules), a chloroplast, and cytoplasmic haematochrome droplets are present in each zoospore. Pyrenoids and eyespots are absent. Flagellar insertion is characterized by “reversed bilateral symmetry”; and zoospores with both right-handed and left-handed arrangements are produced. The ultrastructure of the zoospores clearly indicates that: 1) the mode of flagellar insertion: 2) morphology, number, and arrangement of multilayered structures, and 3) bilaterally keeled flagella are characteristic of the Chroolepidaceae.