Extrachromosomal Circular DNA from Nuclear Fraction of Higher Plants

To determine the cellular location of extrachromosomal circularDNA in higher plants, wheat (Triticum aestivum) and tobacco(Nicotiana tabacum) nuclei were isolated and purified, and theircircular DNAs were examined by electron microscopy. Covalentlyclosed circular (ccc) DNAs were found in nuclear fractions fromboth species. They showed a heterogeneous size distributionranging from 0.1 µm to more than 5 µm in contourlength, with a mean of 1.7 µm (5.3 kbp) for T. aestivumand 1.5 µm (4.7 kbp) for N. tabacum, respectively. Thisdistribution is significantly different from that for smallcircular DNAs in mitochondria. Small polydisperse circular (spc)DNA/protein complexes were also observed by a rapid microscalemethod of mica-press-adsorption for electron microscopy. Complexesof spcDNA/protein showed a similar size distribution to cccDNA.The average number of spcDNA/protein complexes per nucleus wasestimated at more than hundred. The origin and biological functionsof the nuclear circular DNAs are discussed. (Received February 13, 1985; Accepted August 8, 1985)     

Leaf Interveinal Distances Corresponding to Anatomical Types in Grasses

Considerable variation in the interveinal distances in leaveswas observed in both C₃ and C₄ grass species. In C₄ species,interveinal distances of the Mestome Sheath type (89.0 µm)were smaller than those of the Parenchyma Sheath type (140.0µm). In contrast, non-Kranz type (C₃ species) had thelargest interveinal distances of all the anatomical types. Variationin the interveinal distances of leaves of grasses is discussedin relation to photosynthetic ability and water use. (Received August 7, 1984; Accepted January 16, 1985)     

Swelling of Etiolated Oat Protoplasts Induced by cAMP and Red Light

Etiolated oat protoplasts were treated with dibutyryl cAMP tostudy possible function of cAMP in the development by measuringthe protoplast swelling. The mean diameter of protoplasts inthe absence of any chemical treatment was 33.58±1.26(SE) µm, which increased to 36.96±0.86 µmin the presence of 100 µM dibutyryl cAMP. Prostacyclin,a potent activator of adenyl cyclase, also showed a significantswelling effect (diameter 38.01±0.98 µm). Red lightalso elicited the swelling of protoplasts (40.26±0.8µm). ¹Present address: Department of Biology, Pusan National University,Pusan 607, Korea. ²Present address: Department of Horticulture, Cheju NationalUniversity, Cheju 590, Korea. ³Present address: Department of Biological Sciences, Texas TechUniversity, Lubbock, TX 79409, U.S.A. (Received June 29, 1985; Accepted November 18, 1985)     

Isolation and Characterization of Photosynthetically Active Cells from Submersed and Floating Leaves of the Aquatic Macrophyte Potamogeton nodosus Poir

Photosynthetically active cells were isolated by enzymic digestionof floating and submersed leaves of the heterophyllous aquaticmacrophyte Potamogeton nodosus Poir. The yields of cells isolatedfrom floating leaves represented approximately 25% of the leafprotein or chlorophyll, while cell yields from submersed leaveswere only 3%. Photosynthetic activity was maximal in cells isolatedfrom submersed leaves 10 to 14 days after germination of thewinterbuds. Floating leaves were induced by treatment of theplants with abscisic acid. Cells from induced floating leavesshowed maximum photo synthetic rates between 9 and 21 days posttreatment.Phosphoglycerate, fructose 1,6-bisphosphate, sulfate and phosphatewere without significant effect on photosynthesis in eithercell type indicating that the cells were substantially intact.Half-saturation of photosynthesis for bicarbonate was at 0.6mM (pH 7.6) for cells from both leaf types, and the maximumrate was greater for cells from floating leaves. The light intensityfor half-saturation of photosynthesis was approximately 95 µEm^–2s^–1 for cells from both leaf types, and the maximumrate was greater for cells from floating leaves. (Received September 19, 1984; Accepted December 6, 1984)     

Size-selective grazing of bacteria by Bosmina longirostris--an image-analysis study

Image analyses on the filtering apparatus of Bosmina longirostricshowed that the filter mesh is finer on the gnathobasic filterplates of the second and third trunk limbs (ranges from 0.43to 0.97 µm) and coarser for the outer ones of the thirdlimb (ranges from 0.5 to 1.36 µm), and the intersetulardistances increase with body length. Grazing experiments combinedwith image analysis confirmed the efficient grazing of B.longirostrison natural bacteria with cell lengths equal to or larger thanthe intersetular distances of the gnathobasic filter plates.During the experiments, the animals minimized the average celllength of the bacterioplankton assemblages from 0.77–0.96µm to 0.55–0.68 µm, corresponding to the meanof the filter mesh size on the fine gnathobasic filter platesof the experimental populations. The clearance rates for large,elongated or dividing cells with maximal lengths of 0.88–8.40µm were 2.3–17.7 times higher than those for smallsingle coccoids with a diameter of <0.45 µm. The resultsprovide evidence of a significant differential impact of B.longirostrison the bacterial community structure with respect to the shapeand size of the cells, and demonstrate that the species is amore effective bacterial feeder than considered previously.     

Localization of Thermo-Osmotically Active Partitions in Young Leaves of Nuphar lutea

The submerged roots and rhizomes of the aquatic vascular macrophyteNuphar lutea (L.) Sm. are aerated, at least in part, by pressurizedventilation. Depending on temperature differences of up to 5K between the inside of young, just-emerged leaves and the surroundingair, pressure differences of 79 to 100 Pa higher than atmosphericare detectable inside the lacunuous spongy parenchyma of theleaf blades. The pressurization is a consequence of structuralfeatures of leaf tissues separating the air filled spaces ofthe spongy parenchyma from the atmosphere. These tissues areacting as thermo-osmotic partitions. Whereas the dimensionsof the stomatal openings (about 5·6 x 2·4 µm)and of the intercellular spaces of the palisade parenchyma (diametersabout 15 µm) are too large, those of the monolayers ofcells separating the palisade and the spongy parenchyma (diameters:0·7–1·2 µm) are small enough to impedefree gaseous diffusion. This inner non-homogeneous partitioninggives rise to the so-called Knudsen diffusion, a physical phenomenonleading to pressurization of the warmer air inside the spongyparenchyma. The rising pressure difference is strong enoughto establish an air flow through the aerenchyma of the wholeplant and out of the most porous older leaves in which a temperatureinduced pressurization is never detectable. These thermo-osmoticallyactive leaves enhance the influx of air to the rhizome and thediffusion path for oxygen to the roots is shortened to the distancebetween rhizome and root tips. Therefore, pressurized ventilationin Nuphar is seen to be of considerable ecological importancefor plant life in anaerobic environments. Key words: Aeration, leaf anatomy, thermo-osmosis of gases, Nuphar lutea     

THE ULTRASTRUCTURE OF CILIATED CELLS FROM THE SIPHON OF SOLEN CAPENSIS (MOLLUSCA, BIVALVIA)

The structure of ciliated cells from the siphon of Solen capensishas been studied by both scanning and transmission electronmicroscopy. Two types of ciliated cell, based on the numberand length of cilia have been described which resemble thosedescribed in Donax. Type I is characterized by having 26–57({macron}= 43, n = 50) cilia which are 2.5 µm in length;Type II has fewer cilia (5–10; {macron}= 7) which are5 µn long. Both are primary receptors. Estimations ofabundance show that receptors are most numerous on the tipsof the siphon tentacles (8.8 x 10³/mm²). (Received 15 January 1985;     

Regeneration of Tissues in Wounded Stems: A Quantitative Study

When a dicotyledonous stem is wounded by longitudinally splittinga young internode into halves, cells near the cut surface proliferateto form a callus within which vascular tissues differentiateand tend to restore a vascular cylinder in each half. Threephases of regeneration after wounding were identified and quantifiedin stems of three Solanaceous species. (1) In an initial ‘lag’phase, lasting about 2 d, neither cell division nor enlargementwere detected, but mitotic figures were observed within about300 µm of the cut surface. (2) Throughout a second, ‘division’phase, from about days 2–10, cell division and enlargementoccurred. Both were initiated mainly in the two cell layersnearest the surface. A mass of callus formed, with new cellwalls mostly parallel to the surface. Cell enlargement laggedbehind cell division for the first few days, so that mean radialcell diameter decreased until day 6, thereafter remaining almostconstant at 30–40 µm. Towards the end of this phase,mitoses ceased within the callus except in the positions ofthe future vascular and cork cambia, where radial cell diameterfell towards a constant 15–20 µm. (3) During a third,‘differentiation’ phase, cell division was restrictedto the cambial zones, and derivatives differentiated into cork,phloem or xylem according to position. The rate of increasein cell number per transect was 1.5–2.0 cells d^–1,of which more than half was xylem. Capsicum annuum L., sweet pepper, Lycopersicon esculentum Mill., tomato, cambium, cell division, differentiation, regeneration, wounding of stems, xylem     

Size-fractionated phytoplankton carboxylase activities in the Indian sector of the Southern Ocean

During the ANTARES 3 cruise in the Indian sector of the SouthernOcean in October–November 1995, the surface waters ofKerguelen Islands plume, and the surface and deeper waters (30–60m) along a transect on 62°E from 48°36'S to the iceedge (58°50'S), were sampled. The phytoplankton communitywas size-fractionated (2 µm) and cell numbers, chlorophyllbiomass and carbon assimilation, through Rubisco and ß-carboxylaseactivities, were characterized. The highest contribution of<2 µm cells to total biomass and total Rubisco activitywas reported in the waters of the Permanent Open Ocean Zone(POOZ) located between 52°S and 55°S along 62°E.In this zone, the picophytoplankton contributed from 26 to 50%of the total chlorophyll (a + b + c) with an average of 0.09± 0.02 µg Chl l^–1 for <2 µm cells.Picophytoplankton also contributed 36 to 64% of the total Rubiscoactivity, with an average of 0.80 ± 0.30 mg C mg Chla^–1 h^–1 for <2 µm cells. The picophytoplanktoncells had a higher ß-carboxylase activity than largercells >2 µm. The mixotrophic capacity of these smallcells is proposed. From sampling stations of the Kerguelen plume,a relationship was observed between the Rubisco activity perpicophytoplankton cell and apparent cell size, which variedwith the sampled water masses. Moreover, a depth-dependent photoperiodicityof Rubisco activity per cell for <2 µm phytoplanktonwas observed during the day/night cycle in the POOZ. In thenear ice zone, a physiological change in picophytoplankton cellsfavouring phosphoenolpyruvate carboxykinase (PEPCK) activitywas reported. A species succession, or an adaptation to unfavourableenvironmental conditions such as low temperature and/or availableirradiance levels, may have provoked this change. The high contributionof picophytoplankton to the total biomass, and its high CO₂fixation capacity via autotrophy and mixotrophy, emphasize thestrong regeneration of organic materials in the euphotic layerin the Southern Ocean.     

Hormonal Control of Elongation of Tobacco Cells Derived from Protoplasts

Spherical protoplasts (30–40 µm in diameter), isolatedfrom cultured tobacco cells, elongated rapidly during a cultureperiod of 3 to 8 days and finally assumed a long cylindricalform (300–400 µm in length). The optimum concentrationof a-naphthaleneacetic acid and benzyladenine for the elongationwas 0.1 mg/liter and 1 mg/liter respectively. (Received August 10, 1982; Accepted November 12, 1982)     

Nitrate Reduction by Cassava

Nitrate reductase (NR) was assayed in vivo in cassava (Manihotesculenta Crantz). Activity in the leaves ranged from 0 to 2.51µmole of NO₃^– reduced g^–1 h^–1, withno activity in the younger leaves (leaf 1 on top). NR activitywas localized in the sides and toward the tip of the lobes ofthe leaf. (Received December 10, 1985; Accepted April 8, 1986)     

Are bacteria an important food source for rotifers in eutrophic lakes?

In situ grazing measurements using fluorescent particles of0.5, 2.4 and 6.3 µm diameter in eutrophic Lake Loosdrecht(The Netherlands) showed that Anuraeopsis fissa, a small rotifer,filtered the smallest, bacteria-sized particles as efficientlyor more efficiently than the larger particles. In contrast,three other rotifer species (Brachionus angularis, Filinia longisetaand Pompholyx sulcata) filtered the bacteria-sized particlesless efficiently than the larger particles. Both Keratella cochlearisand Conochilus unicornis only ingested the bacteria-sized particles.Anuraeopsis fissa had a higher uptake of fluorescent bacteria-sizedparticles than K.cochlearis, both in 1 µm filtrate oflake water and in lake water. Within both species, uptake didnot differ between juveniles and adults. When cultured on threedifferent size fractions of lake water (1, 3 and 15 µmfiltrate) in July, all rotifer species declined in numbers onthe 1 and 3 µm filtrates, while A.fissa and B.angularisincreased in numbers on the 15 µm filtrate. The high abundanceof small bacteria in the lake water could not support rotiferpopulations. It is concluded that bacteria are not a suitablefood source of high quality for A.fissa because its populationdoes not grow even though the bacterial concentration was higherthan its estimated threshold food concentration. In August,when individually cultured, the mortality was high for all species,but especially for F.longiseta. The lifespan of K.cochleariswas reduced in the 1 and 3 µm filtrates of lake water,compared with in the 15 µm filtrate. The lifespan of A.fissawas similar in all filtrates, but reproduction was reduced inthe 1 and 3 µm filtrates, as in Keratella. On the 15 µmfiltrate, their ages at first reproduction and growth ratesdid not differ. Individuals of A.fissa older than 4 days showeda higher survival in the 15 µm filtrate than in the othertwo filtrates, as did K.cochlearis throughout its life. Hence,bacteria seem to be a more important food source for youngerindividuals of A.fissa than of K.cochlearis.     

Cortical Bundles in the Persistent, Photosynthetic Stems of Cacti

We examined 62 species in 45 genera of the cactus subfamilyCactoideae; all had collateral cortical bundles that permeatedthe broad, water-storing inner cortex and extended to the baseof the outer, photosynthetic palisade cortex. Mean distancebetween cortical bundles was 0.75 mm, similar to the mean spacing(0.74 mm) of veins in leaves of Pereskia, a genus of relictleaf-bearing cacti. In 16 species, both young and extremelyold stem cortex was available for study: in all of these, olderbundles had larger amounts of phloem than did younger bundles,indicating that phloem had been produced for many years. Inten species, older bundles also had more xylem than youngerbundles. In two genera (Rhipsalis and Selenicereus) there werecaps of primary phloem fibres, and in a single species (Pilosocereusmortensenii) cortical bundle xylem contained libriform fibres.All cortical bundle tracheary elements were narrow (radius range,0.91–8.2 µm; mode, 1.8–2.7 µm), similarto Pereskia leaf vein elements (radius range, 1.8–2.7µm); this was much narrower than stem wood vessels (radiusrange, 10–42 um; mode, 23–28 µm). Longitudinalconduction of water and nutrients probably occurs predominantlyin stem wood, with cortical bundles maintaining the broad, voluminouscortex, the outer part of which is the plant's photosynthetictissue and the inner part of which stores water and starch.The cortex of the Cactordeae contains numerous leaflike characters;homeotic genes may be involved in its morphogenesis. Cactaceae, cortical bundles, homeotic, xylem, phloem, evolution     

A method for isolating adult and neonatal airway smooth muscle cells and measuring shortening velocity

Methods are described for isolating smooth muscle cells from thetracheae of adult and neonatal sheep and measuring the single-cell shortening velocity. Isolated cells were elongated,Ca²⁺ tolerant, and contractedrapidly and substantially when exposed to cholinergic agonists, KCl,serotonin, or caffeine. Adult cells were longer and widerthan preterm cells. Mean cell length in 1.6 mMCaCl₂ was 194 ± 57 (SD) µm(n = 66) for adult cells and 93 ± 32 µm (n = 20) for preterm cells(P < 0.05). Mean cell width at thewidest point of the adult cells was 8.2 ± 1.8 µm(n = 66) and 5.2 ± 1.5 µm(n = 20) for preterm cells(P < 0.05). Cells were loaded into aperfusion dish maintained at 35°C and exposed to agonists, andcontractions were videotaped. Cell lengths were measured from 30 videoframes and plotted as a function of time. Nonlinear fitting of celllength to an exponential model gave shortening velocities faster thanmost of those reported for airway smooth muscle tissues. For a sampleof 10 adult and 10 preterm cells stimulated with 100 µM carbachol,mean (± SD) shortening velocity of the preterm cells was notdifferent from that of the adult cells (0.64 ± 0.30 vs. 0.54 ± 0.27 s¹, respectively), butpreterm cells shortened more than adult cells (68 ± 12 vs. 55 ± 11% of starting length, respectively;P < 0.05). The preparative andanalytic methods described here are widely applicable to other smoothmuscles and will allow contraction to be studied quantitatively at thesingle-cell level.

     

Particle capture by Favella sp. (Ciliata, Tintinnina)

Particle capture by the tintinnid ciliate, Favella sp., wasinvestigated with high-speed video microscopy. Experiments withmicrospheres and algal cells indicate that within the size range1–19 p.m. behavioral responses are partly responsiblefor the differential capture of large over small particles.For example, 4 µm microspheres or Isochrysis [spheiicaldiameter (ESD) - 4.5 µ are only captured if they are encounteredin the inner zones of the encounter area. Ten micrometer microspheresor Heterocapsa(14–19 – are captured with a greaterefficiency than smaller particles from the inner zones, butcan also be captured from the outer zone of the encounter area.Capture of particles from the outer zone is associated withcilialy reversal. Microsp1ieres and algal cells of similar sizeare captured in a similar manner.     

Anatomical considerations related to photosynthesis in cotton (Gossypium hirsutum L.) leaves, bracts, and the capsule wall

Light and electron microscopy was used to relate histologicaland ultrastructural differences of cotton (Gossypium hirsutumL.) leaves, bracts, and capsule walls to their different photosyntheticactivities. Light microscopy revealed that the leaf thicknesswas approximately 152µm, had a well-defined internal organizationwith elongated palisade mesophyll cells and loosely packed spongymesophyll cells. In contrast, the bract was thinner (111 µm),lacked a defined palisade layer, and was largely composed ofinternal air spaces. The capsule wall was very thick (1013µm)and composed of numerous tightly packed, paren-chymatous corticalcells with little or no intercellular air space. Chloroplastswith well-defined granal stacks and extensive stroma lamellaewere observed in each of these three tissues, however, theirdensity was always greater in the palisade cells of the leafcompared to spongy mesophyll cells of the bract and the parenchymatouscells of the capsule wall. The low rates of photosynthesis inthe bracts and the capsule wall were associated with the internalorganization of these tissues. Key words: Cotton, photosynthesis, anatomy, cuticle, tissues     

Ultrastructure of Protein Crystals in Potato and Tomato Trichomes

Large protein crystals were located in the leaf and stem trichomesof Solanum tuberosum L. and Lycopersicon esculentum Mill. Inpotato the crystals ranged from 1.05 to 4.5 µm (average2.3 µm) on a side and in tomato they ranged from 1.16to 3.5 µm (average 2.7 µm) on a side. The proteinnature of the crystals was determined by histochemical stainingwith Coumassie brilliant blue R250 and aniline blue black. Thecrystalline structure of the inclusions was observed in ultrathinsections using electron microscopy. In potato, in cleared areasof the cytoplasm, ribosomes were observed scattered among proteinfilaments. The filaments were approximately 7 nm in diameter.Morphologically similar crystals were observed in the tomatotrichomes but the protein filaments were smaller (approximately4 nm in diameter). Protein crystals were also observed in palisadeand spongy parenchyma and epidermal leaf cells in tomato. Protein crystals, trichomes, potato, Solanum tuberosum L., tomato, Lycopersicon esculentum Mill., ultrastructure     

Identification of the Excitable Cells in the Petiole of Mimosa pudica by Intracellular Injection of Procion Yellow

Excitable cells in the petiole of Mimosa pudica were locatedby microelectrode technique and stained with Procion YellowMx4R which was previously filled in the electrode and injectediontophoretically into the cells. Microscopic observations ofsections of the stained petioles revealed that protoxylem parenchymacells and narrow phloem cells were excitable. The protoxylemlocalized just inside the metaxylem was composed almost entirelyof the parenchyma cells which were 106.3±5.2 µmlong (mean±EM, n=15) and 14.2±0.6 µm indiameter (n =33). The excitable phloem cells were 76.4±4.1µm long (n=7) and 7.0±0.3 pan in diameter (n=37)and were thought to be companion cells or narrow parenchymacells or both. Amplitudes of action potentials recorded fromthe petiolar surface had a linear relation to those from theexcitable cells in the same petiole. From this fact and thearrangement of excitable cells in the petiole, we conclude thatwhen the transmission of action potential takes place in thepetiole all excitable cells in it are activated. ¹ Present address: 1st Department of Physiology, Hamamatsu UniversitySchool of Medicine, Handa-cho 3600, Hamamatsu 431-31, Japan. (Received September 7, 1982; Accepted November 8, 1982)     

Drought effects on cellular and spatial parameters of leaf growth in tall fescue

The effect of drought and recovery on cellular and spatial parametersof the growth process in tall fescue leaves was studied in twoexperiments. In both experiments plants grown on vermiculiteand maintained in a controlled environment were submitted toa 7 d drought period generated by withholding water. Droughtwas followed by a 3 d recovery period in experiment II. As leafelongation rate (LER) decreased during developing drought boththe growth zone length (initially 40 mm) and the maximum relativeelemental growth rate (initially 0.09 mm mm^–1 h^–1during the dark period of diurnal cycles) within the growthzone declined. But the growth zone still exhibited a lengthof approximately 15 mm when LER approached 0 under severe drought(–2.0 MPa predawn leaf water potential). The growth potentialof the basal 15-mm-long portion of the leaf was conserved duringthe period when drought effected the complete arrest of leafelongation. A (retrospective) analysis of the position-timerelationships of epidermal cells identified on leaf replicas(experiment II) indicated that the cell flux out of the growthzone responded very sensitively to drought. Before drought theflux was maximum at approximately 3.2 cells (cell file h)^–1during the dark period. Flux decreased to 0 when leaf elongationstopped. Flux also varied diurnally both under well-wateredand droughted conditions. In well-watered conditions it wasabout 30% less during the light than the dark period. Cell elongationwas also sensitive to drought. Under well-watered conditionsepidermal cell elongation stopped when cells attained a lengthof approximately 480 µm. During developing drought cellsstopped elongating at progressively shorter lengths. When LERhad decreased to almost nil, cells stopped elongating at a lengthof approximately 250 µn. When drought was relieved followinga 2 d complete arrest of leaf elongation then cells shorterthan 250 µm were able to resume expansion. Following rewateringcell flux out of the growth zone increased rapidly to and abovethe pre-drought level, but there was only a slow increase overtime in the length at which cell elongation stopped. About 2d elapsed until the leaf growth zone produced cells of similarlength as before drought (i.e. approximately 480 µm). Key words: Epidermal cell length, cell flux, (leaf) growth zone, leaf elongation rate, relative elemental growth rate, position-time relationships (path line, growth trajectory), drought, water deficit     

Colony growth and evidence for colony multiplication in Phaeocystis pouchetii (Prymnesiophyceae) isolated from mesocosm blooms

Using well plates of Phaeocystis pouchetii colonies isolatedfrom experimental mesocosms in western Norway, increases incolony size and division were documented. Median longest lineardimensions increased 0–7 µm h^–1; literaturePhaeocystis globosa values are 0.9–4.7 µm h^–1.Ten to twelve percent of colonies divided at rates of 0.21–0.28divisions day^–1. Daughter colonies were 100 µm smallerthan mother colonies. Colonies delayed 3.5–4.9 days tofirst division, compared with literature values of 4–5days for P. globosa. This study provides the first experimentalevidence for colony division of wild P. pouchetii.