Light and electron microscopical studies onHafniomonas gen. nov. (Chlorophyceae,Volvocales), a genus resemblingPyramimonas (Prasinophyceae)

Based on light and electron microscopical studies ofPyramimonas reticulata the genusPyramimonas is shown to contain a number of unrelated flagellates.P. reticulata andP. montana are transferred to the new genusHafniomonas, cells of which differ fromPyramimonas in shape, in the absence of scales and hairs on the body and flagellar surfaces, in details of the chloroplast, the position of the nucleus, the Golgi apparatus, the internal structure of the flagellar apparatus, and in cell division. The prasinophytePyramimonas contains a characteristic association of a large microbody and a rhizoplast, situated on the nuclear surface. A similar association is being found in an increasing number of prasinophycean flagellates, but is absent inHafniomonas, which is considered related to chlorophycean rather than prasinophycean flagellates. The phylogenetic position ofHafniomonas is discussed, based in particular on details of the unique flagellar apparatus.     

Description of Pyramimonas diskoicola sp. nov. and the importance of the flagellate Pyramimonas (Prasinophyceae) in Greenland sea ice during the winter–spring transition

Pyramimonas Schmarda is a genus of unicellular green flagellates, recorded in marine water and sea ice samples. Pyramimonas is within the prey size range of the most important protozoan grazers in Disko Bay, West Greenland, where this study took place. Despite the potential ecological importance, little is known about the occurrence of the genus. The aim of this study was to explore the biomass of Pyramimonas in developing stages of sea ice and in the water column. Pyramimonas colonized the early stages of sea ice, and the highest percent of Pyramimonas biomass was found in grease ice. The biomass of Pyramimonas was more than a magnitude higher within sea ice compared to the surface water. The results illustrate that Pyramimonas from the ice is an important contributor to the plankton community prior to the spring bloom. An undescribed species, Pyramimonas diskoicola sp. nov., was found. Based on morphology and ultrastructure, combined with molecular phylogeny inferred from the small-subunit SSU rDNA and the large-subunit chloroplast-encoded rbcL, the species was placed in subgenus Vestigifera. The cells possessed four flagella, measured 8.3 ± 2.6 μm in length and 5.1 ± 0.8 μm in width, and were characterized by an uplifted quadrant in the center of the box scales, not seen at any other Pyramimonas species. The phylogenetic analyses indicated P. diskoicola to be closely related to other polar sea ice species of Pyramimonas.     

Neue Arten der Gattung Pyramimonas Schmarda

Zusammenfassung Es wird auf eine grosse Formenfülle der Gattung Pyramimonas hingewiesen. Mehrere neue Arten and eine neue Varietät, die in Nordmähren (Tschechoslowakei) gefunden werden, werden erwähnt und abgebildet. Folgende Arten werden beschrieben: Pyramimonas recta, P. fasciata, P. radiata, P. conradii, P. cribrum, P. polychloris, P. rapa, P. pluristigma, P. fabelliformis und die Varietdt P. reticulata var. conica.

---

Summary The great number of Pyramimonas-species is discussed. Some new species, found in Nordmähren (Czechoslovakia) are described and pictured. The new species described are: Pyramimonas recta, P. fasciata, P. radiata, P. conradii, P. cribrum, P. polychloris, P. rapa, P. pluristigma, P. fabelliformis and the variety P. reticulata var. conica.

     

THE ULTRASTRUCTURE OF PYRAMIMONAS PSEUDOPARKEAE SP. NOV. (PRASINOPHYCEAE) FROM SOUTH AFRICA1

The ultrastructure of Pyramimonas pseudoparkeae sp. nov., a member of the class Prasinophyceae occurring in tidal pools along the east, south and west coast of South Africa, is described. The cell surface is covered by three distinctive body scales whilst the flagellar surfaces possess four types of scales. The structure of these scales is described. P. pseudoparkeae resembles Pyramimonas parkeae Norris and Pearson but differs in the structure of the type 2 body scale. The symmetry and ultrastructure of the cell are described with special attention given to the flagellar apparatus. Preliminary information on the life cycle of this species is presented. This new species is compared with other closely related members of the genus Pyramimonas.     

Diversity of swimming behavior in Pyramimonas (Prasinophyceae)

High‐speed videography and inverted light microscope observations of cultures of several Pyramimonas Schmarda species (Pyramimonas chlorina Sym et Pienaar, Pyramimonas disomata McFadden Hill et Wetherbee, Pyramimonas gelidicola McFadden Wether‐bee et Moestrup, Pyramimonas mantoniae Moestrup et Hill, Pyramimonas melkonianii Sym et Pienaar, Pyramimonas mitra Moestrup et Hill, Pyramimonas moestrupii McFadden, Pyramimonas mucifera Sym et Pienaar, Pyramimonas nephroidea McFadden, Pyramimonas orientalis McFadden Hill et Wetherbee, Pyramimonas parkeae Norris et Pearson, Pyramimonas propulsa Moestrup et Hill) revealed swimming behavior to be much more diverse than originally reported for the genus. This diversity shows gradation between extremes of recurved flagellar beating and non‐biphasic ciliary beating. Comparison with the behavior of presumably more primitive members of the Pyramimonadales (Cymbomonas tetramitformis Schiller and Pterosperma cristatum Schiller) leads to the conclusion that the former condition is primitive and the gradation may assist in reconstructing phylogenetic relationships within the genus Pyramimonas, particularly as it is consistent with phylogenies derived from ultrastructural and molecular data.     

Screening for marine nanoplanktic microalgae from Greek coastal lagoons (Ionian Sea) for use in mariculture

Mediterranean mariculture uses imported strains of marine phytoplankton, raising questions of ecological risk and ability to adapt to local conditions for mass culture outdoors. In this context, we report here on the mass-culture potential and chemical composition of six strains of Prasinophyceae (five strains of Tetraselmis sp. and one Pyramimonas sp.) isolated from a Greek coastal lagoon. Proximate composition had a pattern of 10–20% ash, 35–65% protein, 6–10% lipids, and 25–45% other organics including carbohydrates. The amino acid profiles were typical for the marine representatives of the class. All strains had a high PUFA content with dominant the ω3 fraction in four of them. The fatty acid profiles indicated a Tetraselmis strain with high EPA (14%) and a Pyramimonas strain with high DHA (6%). These strains might be a good alternative for the common commercial strains used in Mediterranean aquaculture.     

Observations with the electron microscope on Asteromonas gracilis Artari emend. (Stephanoptera gracilis (Artari) wisl.), with some comparative observations on Dunaliella Sp

A clone of Asteromonas gracilis has been examined in culture by means of light and electron microscopy. The morphological observations of Artari (1913) have been confirmed and extended. New observations include demonstration of an unusual type of pyrenoid somewhat resembling that of Prasinocladus, with tubular channels entering the pyrenoid centre from outside the plastid and penetrated by haustorial outgrowths from the nuclear envelope. The very different structure characteristic of Dunaliella is briefly illustrated. Other comparisons indicate that A. gracilis is the type species and sole known representative of a very distinct genus which cannot be confused with Pyramimonas and is unlikely to be synonymous with the imperfectly known genus, Stephanoptera.     

LIGHT AND ELECTRON MICROSCOPY OF A PUNCTATE SPECIES OF PYRAMIMONAS,P. MUCIFERA SP. NOV. (PRASINOPHYCEAE)1

Pyramimonas mucifera sp. nov., a punctate species of the genus, is unusual both behaviorally and at the fine structural level. It forms two distinct populations in culture, one benthic and one planktonic. Planktonic forms are more conventional for the genus, but benthic forms are found in loosely packed mucilage, have flagellar rather than ciliary beating of the flagella, and display a higher degree of metaboly. Ultrastructurally this species is unusual in that it has a unique scale complement and the cells contain numerous muciferous vesicles, leaving only small pockets of cytoplasm containing the usual organelles. This species has a 3–1 type flagellar apparatus but has an additional fibrillar band, a 4–3-2–3 microtubular root system and a flexible synistosome. The discovery of a mucilage-producing species of Pyramimonas draws attention to possible links with other prasinophytes (Halosphaera) and green algae of questionable affiliation (Oltmannsiellopsis and Hafniomonas). It also provides a model of the primitive pyramimonad.     

A QUANTITATIVE ANALYSIS OF THE MEGAGAMETOPHYTE OF FIVE SPECIES OF CORNUS L.

Comparisons of variations of morphological features, treated quantitatively, of the megagametophytic stages in Cornus asperifolia Michaux., C. stricta Lam., C. amomum Miller, C. florida L., and C. alternifolia L. suggest differences in growth patterns of their megagametophytes. The successful application of the clearing fluid 41/2 (Herr Fluid) affords adequate data for assessment of quantitative features of such minute plants. This method, less time consuming than traditional paraffin sectioning techniques, allows for statistical treatment of a sufficient number of species for valid inferences of angiospermous embryological features. An investigation of megagametophytic size was directed both to successive stages within each species and to each stage among all five species. In both instances, means, confidence intervals (CI), and growth increments (GI) were compared. Without exception, the largest means for length and width occurred in the 8-nucleate stage for all species. The greatest GI for length occurred in the 8-nucleate stage for C. stricta, C. alternifolia, and C. asperifolia and in the 4-nucleate stage for C. florida. The 4- and 8-nucleate GI were nearly equal for C. amomum. Overlapping of 95 % CI between successive stages indicated uniform growth between stages for the megagametophyte. Only C. asperifolia had CI overlapping between all successive stages for length and width. These five species are clearly separated on the basis not only of variations in qualitative features but also on megagametophytic growth patterns among species.     

Chemical Composition and Antimicrobial Activity of Achillea tenuifolia Lam. Essential Oil at Different Phenological Stages from Khoy

Achillea species and in particular Achillea tenuifolia Lam . is generally used as a food flavor and traditional remedies, especially in the initial developmental stage for medical conditions in the Mediterranean part of Iran. In this report, we extracted the essential oil from the aerial parts of A. tenuifolia (collected from Khoy), at various developmental stages (i. e., vegetative, flowering and fruiting), characterized them and studied their antibacterial activities. Of 46, 51 and 38 components found in the vegetative, flowering, and fruiting stages, respectively, 35 were present in all three stages, including oxygenated terpenes such as carvacrol (30.85–34.11), germacrene C (16.21–17.87), spathulenol (7.26–8.96), β‐sesquiphellandrene (4.11–4.25), τ‐muurolol (2.27–3.25) and α‐cadinol (2.01–3.29). We witnessed that the composition of the essential oils varies with phenological stages and geographic regions. The essential oil demonstrated substantial antibacterial properties against both Gram‐positive and Gram‐negative bacteria, indicated by disk method, Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. Except Pseudomonas aeruginosa, the essential oils of various phenological stages showed higher antibacterial activity against tested bacteria, with Bacillus anthracis as the most sensitive strain. Moreover, although antibacterial characteristics of the essential oil from the vegetative and flowering stages were similar (p=0.91), they were significantly different from those of fruiting stage (p<0.005 in both MIC and MBC tests). This emphasizes the importance of the developmental stage of the plant in the biological properties of its essential oil and justifies the widespread application of this plant in the vegetative stage.     

ABSOLUTE CONFIGURATION ANALYSIS OF THE FLAGELLAR APPARATUS OF PTEROSPERMA CRISTATUM (PRASINOPHYCEAE) AND CONSIDERATION OF ITS PHYLOGENETIC POSITION1

Pterosperma cristatum Schiller, a member of the Pra-sinophyceae, was examined with light and electron microscopy with special attention on the absolute configuration of flagellar apparatus components and associated structures. This alga is characterized by asymmetrically arranged basal bodies, connecting fibers and microtubular roots. The microtubular root system is homologous with the cruciate root system, the so-called X-2-X-2 root system typical of non-charophycean green algae. Two ducts are associated with microtubular roots. A similar flagellar apparatus and duct system was found in two other prasinophyte genera, Pyramimonas and Halosphaera. The close phylogenetic affinity of these three genera is discussed.     

Protein-protein interactions of the light-harvesting chlorophyll protein. II. Evidence for two stages of cation independent association

In a previous paper, we observed a two-stage cation-independent association of the light-harvesting chlorophyll protein from spinach chloroplasts based on concentration-dependent changes in the sedimentation coefficient. The two stages of association occurred between (2–4) and (4–7) μg/ml chlorophyll. In this paper, we provide further evidence for this association.This includes: (1) A decrease in the number of divalent cation binding sites in the second stage of association. (2) A corresponding decrease in the extent of the cation-dependent association. (3) A positive deviation from Beer's law for chlorophyll b for both stages of the cation-independent association and a positive deviation for chlorophyll a for the second stage of association only. (4) A change in the fluorescence emission of both chlorophyll a and b. The change for chlorophyll b was observed for both steps of association whereas that for chlorophyll a was observed for the second step of association only. Therefore, the first stage of association affects only chlorophyll b whereas the second stage alters the environment of both chlorophyll a and b. (5) In addition, divalent cations quenched chlorophyll fluorescence. However, the quenching which required 200–300 μM divalent cation for half-maximal effects was related neither to divalent cation binding nor to the divalent cation-induced association of the protein.     

不同生长季节黑果枸杞的根际细菌群落结构

【目的】黑果枸杞是一种耐盐植物,是我国西北干旱区盐渍土改良的优良植物物种,其根际土壤细菌群落结构在不同生长时期的变化特征尚不清楚。【方法】本研究采用Illumina MiSeq高通量测序研究了黑果枸杞3个生长阶段的根际土壤细菌群落结构的动态变化。【结果】所有样品中共获得317467条序列,对应于7028个细菌/古细菌OTUs。根际土壤细菌群落的α多样性显著高于非根际土壤。衰老期根际细菌的多样性和丰富度明显低于营养生长期和花/果期。变形菌门和酸杆菌门的相对丰度随生长时期的演变而逐渐降低,而蓝细菌门则相反。厚壁菌门的丰度在衰老期明显高于营养生长期和花/果期。优势属的组成也随生长期的演变而改变,营养生长期、花/果期、衰老期的优势属数量分别为17、16、4,且组成也具有差异。相似性分析表明营养生长期和花/果期的根际细菌群落具有很高的相似性,衰老期根际细菌群落组成与生长期和花/果期具有很高差异,然而与非根际土壤的群落结构具有较高的相似性。【结论】根际土壤细菌群落多样性和组成随生长期的改变而表现出明显的动态变异性,表明黑果枸杞生长时期对根际土壤细菌群落结构具有重要的影响。     

Leaf anatomy of <Emphasis Type="Italic">Cynara scolymus</Emphasis> L. in successive micropropagation stages

Summary Leaf structure along the successive stages of Early French artichoke Cynara scolymus L. micropropagation was characterized using light and transmission electron microscopy. The mesophyll presents disorganized spongy and palisade parenchyma with large intercellular spaces and a few small chloroplasts in the leaves of plants cultured in vitro. In addition, both epidermal surfaces of such leaves invariably show a cell wall of the same thickness with a very thin cuticle and open stomata. In the root differentiation stage in vitro, structural changes take place in the leaves that are favorable for survival in the acclimatization stage: conspicuous cuticle, greater cell wall thickness, functional stomata, better mesophyll organization, developed vascular bundles, and the presence of sclerenchymatous tissue are observed. These features found in later in vitro stages are maintained in the following ex vitro stages, some becoming more evident. Our results demonstrate that the structural changes required to ensure appropriate acclimatization of micropropagated artichoke plants begin at the root differentiation stage, which can reduce in vivo acclimatization time and achieve greater survival of transferred plants.     

Sublethal effect of chlorpyriphos and fenpropathrin on functional response of Habrobracon hebetor (Hym.: Braconidae)

One of the criteria for selecting a suitable natural enemy is its functional response to different densities of the host. Habrobracon hebetor (Say) is a larval ectoparasitoid of several species of lepidopteran. Functional response of H. hebetor, after treatment with LC₂₅ of chlorpyrifos and fenpropathrin in three stages, was assessed to different densities of fifth instar of Ephestia kuehniella (2, 4, 8, 16, 32 and 64). LC₂₅ values were estimated to be 0.043, 5.36 and 2.00 and 0.38, 13.9 and 4.45?mg a.i. l^?1 for the larval, pupal and adult stages for chlorpyrifos and fenpropathrin, respectively. Host densities were offered to treated wasps for 24?h. Experiments were replicated 10 times. The results showed that the functional responses of wasps in control as well as immature stages treatments were Type III, whereas those of adult stage were Type II. a (Type II) or b (Type III) and T_h parameters were evaluated with Roger’s equation. b and T_h in control were assessed to be 0.0129 and 0.1723?h, respectively. These parameters were estimated in chlorpyrifos and fenpropathrin to be 0.0052, 0.00718 and 0.1945, 0.2492?h in larval stage; 0.00817, 0.00916 and 0.2736, 0.3108?h in pupal stage; and a and T_h were 0.0064, 0.0137?h^?1 and 0.2492, 0.2242?h in the adult stage, respectively. The insecticide had no significant effect on the searching efficiency of H. hebetor compared with those treated in control; however, they had significant effect on the handling time of the parasitoid and increased it, except in the larval stage treated with chlorpyrifos treatment.     

Temporal relationships betweenZea mays,ostrinia nubilalis (Lep.: Pyralidae) and endophyticBeauveria bassiana

The entomopathogenic fungus,Beauveria bassiana (Balsamo) Vuillemin, was applied to whorl-stage (V7) corn,Zea mays L., by foliar application of a granular formulation of corn grits containing conidia or by injection of a conidial suspension. All plants were infested with European corn borer larvae,Ostrinia nubilalis (Hübner), at the V7 (whorl), V12 (late-whorl), or V17 (pretassel) stage of plant development. Plants infested at whorl and late-whorl stages had significantly more European corn borer tunneling than did plants infested at the pretassel stage. The percentage of plants colonized byB. bassiana did not differ significantly among the whorl, late-whorl, and pretassel stages. As the plants matured,B. bassiana was isolated from different plant areas, with the pith more frequently colonized than the leaf collars. Foliar application ofB. bassiana provided immediate suppression ofO. nubilalis in those plants infested at whorl stage. The reduced efficacy ofB. bassiana at the intermediate plant stages relative to efficacy at harvest is discussed. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Growth and accumulation of storage reserves by somatic embryos of Ocotea catharinensis Mez. (Lauraceae)

The focus of this study was the assessment of in vitro growth of embryogenic cultures of Ocotea catharinensis Mez. (Lauraceae) on Woody Plant Medium (WPM) supplemented with 22.7 g l^–1 sorbitol, 2 g l^–1 Phytagel, 20 g l^–1 sucrose and 400 mg l^–1 glutamine and the biochemical analysis of somatic embryos at different developmental stages (globular, early cotyledonary, cotyledonary and mature). The embryoids underwent repetitive embryo-genesis and developed non-synchronously, throughout the culture period. Dry weight increased 12.6- and 26.8-fold after 3 and 5 weeks, respectively. During the 5 week culture period a reduction in the frequency of embryoids at the globular stage and increasing frequencies of embryoids at early cotyledonary, cotyledonary and mature stages were observed. Embryoids at the mature stages presented a small but significantly higher water content than those at the globular stage. Therefore, embryoid expansion at the latter stages of development was mainly due to water uptake. Embryoids at the globular stages accumulated higher levels of total proteins while those at the cotyledonary and mature stage showed higher levels of soluble sugars and starch (physiological markers of embryo maturation). Thus, significant differences in the profiles of accumulated storage reserves were detected in the embryoids at different developmental stages in the culture conditions tested.     

The larval development of an Australian limnadiid clam shrimp (Crustacea, Branchiopoda, Spinicaudata), and a comparison with other Limnadiidae

The adult morphology of the Australian Limnadopsis shows some remarkable differences to that of other Limnadiidae. These differences are not reflected in its larval development. In Limnadopsis parvispinus, larval development comprises six stages. In stages I and II only the three naupliar appendages are present: the antennule as a small bud, the biramous antenna as the main swimming organ, and the mandible. The antennal protopod bears two endites, the proximal naupliar process and a more distal endite. In stage III a bifid naupliar process (in earlier stages not bifid) and the first signs of the carapace and trunk limb anlagen (undifferentiated rudiments) appear. In stage IV the carapace anlagen become more pronounced. The number of trunk limb anlagens increases to five, and differentiation has commenced. In stage V the first five pairs of trunk limbs are differentiated to varying degrees. The anterior-most four pairs of trunk limbs are subdivided into five endites, a small endopod, an exopod and an epipod. The bivalved carapace covers the anterior-most limbs. In larval stage VI the carapace is larger and the trunk limbs are further differentiated. A general pattern in the sequence of larval stages is the increasing number of sensilla on the antennules. From the last larval to the first postlarval stage, a significant change in morphology takes place. The trunk limbs are now used for swimming. Typical larval organs are much smaller than in the last larval stage. A comparison with other representatives of the Limnadiidae shows a high degree of correspondence, with most differences explained by the heterochronous appearance of characters during development. Five to seven stages are described for all studied Limnadiidae, including one particular stage in which four fully developed setae, a bifid naupliar process and the first signs of carapace anlagen are present. These characters are found in stage III in L. parvispinus, Limnadia stanleyana, Eulimnadia texana, and Imnadia yeyetta but in stage IV in E. braueriana and L. lenticularis. Based on a comparison of the larval stages of six limnadiid and one cyzicid species, we conclude that at least six naupliar stages belong to the limnadiid ground pattern.     

Abortion and determination of stages for embryo rescue in crosses between sweet-potato,Ipomoea batatas Lam. (2n=6x=90) and its wild relative,I. trifida (H. B. K.) G. Don. (2n=2x=30)

Summary The frequency of aborted fruits and the changes and abnormalities that occur during the embryo development in intraspecific crosses of sweet-potato Ipomoea batatas (2n=6x=90) and interspecific crosses between I. batatas and I. trifida (2n=2x=30) were investigated in order to study the causes of the low seed production. Three genotypes of I. batatas and 18 genotypes of I. trifida were intermated. The frequency of aborted fruits was below 25% in the intraspecific crosses and over 90% in the interspecific crosses. Paraffin sections were used to examine the developmental stages of fruits and seeds. Embryos in different developmental stages were observed to determine the stage of abortion. These observations permitted the identification of developmental stages of embryo rescue in interspecific crosses. There were no significant differences in the frequency of embryo abortion before the early globular stage among female sweet-potato progenitors for the intraspecific and interspecific crosses. The frequency of the late occurrence of embryo abortion (when embryo abortion occurs after the pre-globular stage) was higher in interspecific crosses (19.1%) than in intraspecific crosses (5.5%). The frequency of the late occurrence of embryo abortion in interspecific crosses was higher at the globular stage (9.6%) than at the heart stage (4.3%). Providing that embryo rescue is conducted in interspecific crosses, the estimated number of potentially viable embryos could be increased: 30 times with embryos at the globular stage; 20 times with embryos at the heart stage; and 11 times if embryos at the torpedo stage were used for the rescue with respect to the seed set. The results suggested that the appropriate time for embryo rescue in interspecific crosses is at the globular stage. If embryos could be rescued at the globular stage, it would be possible to increase the number of surviving embryos up to 30 times in interspecific crosses and 0.02 times in intraspecific crosses with respect to natural conditions without embryo rescue.This research was initiated during sabbatical of M.I. at the Asian Vegetable Research and Development Center (AVRDC) in Taiwan     

CHLOROPELTA GEN. NOV., AN ULVACEOUS GREEN ALGA WITH A DIFFERENT TYPE.OF DEVELOPMENT1

A diminutive, distromatic ulvaceous green alga was collected in southern California and studied in culture. The initial stages of development resemble those found in the Ulvaceae sensu Bliding. Germlings pass through a uniseriate filamentous stage, a multiseriate stage and a monostromatic saccate stage. At this stage the development departs from the developmental patterns found in the Ulvaceae. Each cell of the monostromatic upright portion of the germling undergoes a single division in a plane parallel to the surface of the germling to form a distromatic saccate germling. Rupture of the apical end of the germling and continued growth eventually results in a peltate distromatic alga superficially resembling Ulva. Based on the developmental pattern, which is unique to the green algae, the new genus Chloropelta gen. nov. and new species Chloropelta caespitosa sp. nov. are proposed for this alga.